V13I3 (Summer 2023)

Innovative VETERINARY CARE

VOLUME 13 ISSUE 3

WHAT GENETICS TELLS US ABOUT LONGEVITY

CONTROLLING DIET, ENVIRONMENT, AND STRESS CAN POSITIVELY IMPACT A DOG OR CAT’S GENES AND LENGTHEN THEIR LIFESPAN. — P. 8

HOW VETERINARY DIAGNOSTICS HAVE ADVANCED IN 50 YEARS

TAKING A TCVM APPROACH TO FELINE COLITIS

SUMMER 2023

www.IVCJournal.com

Veterinary diagnostics, along with disease management and treatment options, have come a long way since the 1970s. — P. 12

By identifying the patterns of disharmony leading to feline colitis, the TCVM practitioner can treat the problem and bring the cat’s body back into balance. —­P. 48

CHRONIC DIARRHEA IN DOGS — WHY AN INTEGRATIVE APPROACH HELPS

WAYS TO SUPPORT THE EQUINE MICROBIOME

An integrative treatment plan that encompasses the interconnected factors contributing to this challenging problem can lead to successful outcomes. — P. 18

EPIGENETICS AND CANINE CANCER — WHAT'S THE LINK?

Understanding the epigenetic changes associated with cancer in dogs provides insights into underlying disease mechanisms and identifies new treatment targets. — P. 26

Why providing forage diversity while minimizing stress will help balance a horse’s microbiome. — P. 54

CREATE A SUSTAINABLE MARKETING STRATEGY FOR YOUR PRACTICE

For optimal success, you need to be a marketer as well as a veterinary professional – here’s what to do. — P. 40

SUMMER 2023 EDITORIAL DEPARTMENT

Editor-in-Chief: Dana Cox Managing Editor: Ann Brightman Associate Editor IVC: Omer Rashid, DVM Senior Content Editor: Ashley Tonkens Senior Content Editor: Belinda Rose-Innes Graphic Design Lead: Ethan Vorstenbosch Graphic Designer: Joy Sunga

COLUMNISTS & CONTRIBUTING WRITERS

Susan Albright, DVM Keri Barron, PhD Chris Bessent, DVM, MSOM, Dipl. OM, LAc Suus Bettink, DVM Ron Carsten, DVM, PhD, CVA, CCRT Todd Cooney, DVM, CVH Kelly Diehl, DVM, MS W. Jean Dodds, DVM Melissa Kellagher, AHVMA Director of Operations Megan Kelly, BVSc, CCRP Cynthia Lankenau, DVM Melissa L. Magnuson, DVM Lisa Miller, DVM, CCRT, CVA Omer Rashid, DVM Margo Roman, DVM, CVA, COT, CPT, FAAO Keith Weingardt, DVM, CVA, CTCVM

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contents FEATURES

LONGEVITY IN 8 INDUCING COMPANION ANIMALS: WHAT GENETICS TELLS US

A SUSTAINABLE MARKETING STRATEGY FOR YOUR 40 CREATING VETERINARY PRACTICE

By Melissa L. Magnuson, DVM

By Megan Kelly, BVSC, CCRP

The genetics of companion animals are affected by diet, environment, and stress. By controlling these three aspects, we care for our patients’ genes and extend their longevity.

As a veterinary professional, you also need to be a marketer — and learn how to measure your marketing efforts for optimal success.

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HOW VETERINARY DIAGNOSTICS HAVE ADVANCED OVER THE LAST 50 YEARS

By Margo Roman, DVM, CVA, COT, CPT, FAAO

Thanks to a range of scientific and clinical advancements, veterinary diagnostics, along with disease management and treatment options, have come a long way since the 1970s.

Used with ozone therapy and other modalities, MBRT led to full recovery for an injured dog with an antibioticresistant urinary tract infection and damage to his autonomic nerves.

By Ron Carsten, DVM, PHD, CVA, CCRT Optimal support for dogs with chronic diarrhea relies on an integrative treatment plan that encompasses the interrelated factors contributing to this challenging health problem.

CLOSER LOOK AT THE LINK 26 ABETWEEN EPIGENETICS AND CANINE CANCER

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By Chris Bessent, DVM, MSOM, DIPL. OM, LAC By identifying the patterns of disharmony leading to feline colitis, the TCVM practitioner can use several modalities to treat the problem and bring the cat’s body back into balance.

INFLUENCING THE EQUINE MICROBIOME — AND 54 FACTORS HOW TO SUPPORT IT

By Suus Bettink, DVM

Understanding the epigenetic changes that occur in canine cancer can provide insights into the underlying mechanisms of the disease, and help identify new targets for treatment.

High stress levels and a lack of forage diversity are among the factors causing imbalances in the equine microbiome.

By Lisa Miller, DVM, CCRT, CVA

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APPROACH TO 48 TCVM FELINE COLITIS

By Kelly Diehl, DVM, MS

THERAPY FOR 32 PBM MUSCULOSKELETAL DISORDERS IN COMPANION ANIMALS

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AUTONOMIC NERVES

By W. Jean Dodds, DVM

INTEGRATIVE APPROACH TO 18 AN CHRONIC DIARRHEA IN DOGS

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HELPS DOG RECOVER 44 MBRT FROM ANTIBIOTIC-RESISTANT INFECTION AND DAMAGED

OA and other musculoskeletal disorders are common in companion animals. Laser or PBM therapy is an alternative treatment option of choice for these patients.

PROFESSIONALS 59 VETERINARY ARE VULNERABLE TO STRESS — 7 WAYS TO EASE THE ANGST

By Omer Rashid, DVM Stress is a serious problem among veterinarians. Recognizing the signs and using these calming techniques will help you control your stress before it becomes overwhelming.

advisory board Dr. Richard Palmquist, DVM GDipVCHM(CIVT) CVCHM (IVAS), graduated from Colorado State University in 1983. He is chief of integrative health services at Centinela Animal Hospital in Inglewood, California, former president and research chair of the AHVMA, and an international speaker in integrative veterinary medicine. Dr. Palmquist is a consultant for the Veterinary Information Network (VIN) and a past president of the AHVM Foundation. He has published two books, one for conventional veterinarians and a second for clients discussing how integrative thinking works.

48 COLUMNS & DEPARTMENTS

7 Editorial 30 Industry Innovations 39 From the AVH

Michelle J. Rivera, MT, VDT, is an instructor at the University of Wisconsin and The Healing Oasis Wellness Center, a post-graduate educational institution offering state-approved programs. She is coowner of The Healing Oasis Veterinary Hospital, offering massage, rehabilitation, chiropractic and Chinese and Western Herbology.

Michelle completed the Chinese Herbal Medicine program from the China Beijing International Acupuncture Training Center, and is certified in Chinese Medicine by the Wisconsin Institute of Chinese Herbology.

Dr. Joyce Harman, DVM, MRCVS, graduated in 1984 from Virginia Maryland Regional College of Veterinary Medicine. Her practice is 100% holistic, using acupuncture, chiropractic, herbal medicine and homeopathy to treat horses to enhance performance and those with a variety of chronic conditions, with an emphasis on Lyme Disease.

Her publications include the Pain Free Back and Saddle Fit Books, and numerous articles in lay and professional magazines. She maintains an informative website: www.harmanyequine.com.

43 From the AHVMA 51 From the VMAA 53 From the VBMA IN THE NEWS:

58 New Endocrine Guidelines from AAHA 62 How Updated State Laws Could

Change Virtual Veterinary Care

Dr. Steve Marsden, DVM, ND, MSOM, Lac. Dipl.CH, CVA, AHG lectures for IVAS, the AHVMA, the AVMA, and numerous other organizations. He is co-founder of the College of Integrative Veterinary Therapies and is a director emeritus of the National University of Natural Medicine in Portland OR. He authored the Manual of Natural Veterinary Medicine (Mosby); and Essential Guide to Chinese Herbal Formulas (CIVT). Dr. Marsden is extensively trained in alternative medicine, including Chinese herbology, acupuncture and naturopathic medicine. He has holistic veterinary and naturopathic medical practices in Edmonton, Alberta. In 2010, Dr. Marsden was named Teacher of the Year by the AHVMA; and Small Animal Veterinarian of the year by the CVMA in 2009.

Dr. Jean Dodds, DVM, received her veterinary degree in 1964 from the Ontario Veterinary College. In 1986, she moved to Southern California to establish Hemopet, the first non-profit national blood bank program for animals. Dr. Dodds has been a member of many national and international committees on hematology, animal models of human disease, veterinary medicine, and laboratory animal science. She received the Holistic Veterinarian of the Year Award from the AHVMA in 1994.

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Dr. Barbara Fougere, DVM, CVAA graduated in 1986, and was named the American Holistic Veterinary Medical Association Educator for 2011. Dr. Fougere is the principal and one of the founders of the College of Integrative Veterinary Therapies. She has continued studying over the last 26 years, and has three Bachelor degrees, two Masters degrees, three post Graduate Diplomas, several Certifications and numerous other courses under her belt.

Dr. Christina Chambreau, DVM, CVH, graduated from the University of Georgia Veterinary College in 1980. She is a founder of the Academy of Veterinary Homeopathy, was on the faculty of the National Center for Homeopathy Summer School and has been the holistic modality adjunct faculty liaison for the Maryland Veterinary Technician Program and is the former Associate Editor of IVC Journal. Dr. Chambreau teaches classes in homeopathy for animals, lectures on many topics, speaks on Radio and TV, and is the author of the Healthy Animal’s Journal among other titles. She is now on the faculty of the Holistic Actions Academy, which empowers members to keep their animals healthy with weekly live webinars.

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contributors 1

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1. CHRIS BESSENT, DVM, MSOM, DIPL. OM, LAC

6. MEGAN KELLY, BVSC, CCRP

2. SUUS BETTINK, DVM

7. MELISSA L. MAGNUSON, DVM

Dr. Chris Bessent has over 30 years of experience in veterinary medicine, and has certificates in veterinary acupuncture, veterinary chiropractic, and veterinary Chinese herbology. She also received her degree in veterinary nutrition and founded The Simple Food Project, formulating freeze-dried raw food diets for cats and dogs using nothing but whole foods. Currently, Dr. Bessent divides her time between The Simple Food Project and Herbsmith, a manufacturer of premium quality supplements for pets. Both are owned and operated out of her facilities in southeastern Wisconsin (simplefoodproject.com, herbsmithinc.com). p.48

Dr. Suus Bettink is a Dutch holistic veterinarian who lives in Portugal. She studied at Faculteit Utrecht and started her career in a rural area where her patients were mainly horses and cows. She switched then to a pharmaceutical company and made her way back to the practice field in a companion animal practice. When following her own internal path, Dr. Bettink started her own company called Praktijk Flip (Practice Flip). She works with quantum energy and herbs. Her special interest and area of knowledge is the gut microbiome.

She combines that with individualized patient plans using herbs and working on stress. p.54

3. RON CARSTEN, DVM, PHD, CVA, CCRT

Dr. Ron Carsten was one of the first veterinarians practicing in Colorado to advocate an integrative approach to patient management. He has lectured on the integrative approach and nutritional supplements, and been published in peer-reviewed journals. Dr. Carsten developed the original Standard Process Veterinary Formulas™. He has an AAS in animal health technology, a BS in microbiology, MS in anatomy and neurobiology, and PhD in cell and molecular biology. He is a certified veterinary acupuncturist and canine rehabilitation therapist, and practices in Glenwood Springs. Dr. Carsten founded Colorado Animal Rescue in Glenwood Springs, and has served on the Colorado State Board of Veterinary Medicine. Dr. Carsten is the 2022 recipient of the Colorado Veterinary Medical Association Distinguished Service Award. p.18

4. KELLY DIEHL, DVM, MS

Dr. Kelly Diehl received her DVM from the University of Tennessee and started her career in an emergency clinic in New Jersey. She completed an internship at the Animal Medical Center in NYC, then completed a residency in small animal medicine at Colorado State University. Dr. Diehl joined the staff of the Veterinary Referral Center of Colorado as co-owner of the internal medicine section. After 14 years, Dr. Diehl left private practice to pursue a career in medical communication and joined Morris Animal Foundation in 2013.

Dr. Diehl is a board-certified small animal internal medicine specialist with an interest in gastroenterology and immunology. p.26

5. W. JEAN DODDS, DVM

Dr. Jean Dodds received her veterinary degree in 1964 from the Ontario Veterinary College. In 1986, she established Hemopet, the first non-profit national blood bank program for animals. Dr. Dodds has been a member of many committees on hematology, animal models of human disease and veterinary medicine.

She received the Holistic Veterinarian of the Year Award from the AHVMA in 1994. p.12

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Dr. Megan Kelly has been a veterinarian for 19 years. After completing a Post Graduate Certificate in Veterinary Rehabilitation at the University of Tennessee, she opened her own practice, Holisticvet, in Diep River, Cape Town. In 2014, Dr. Kelly decided to take a sabbatical from practising and has since been swept up into the online world of veterinary education. She owns and runs onlinepethealth.com, an international continuing education site for veterinary rehabilitation therapists. She creates and provides solutions for vets and veterinary rehab therapists through webinars and online conferences. p.40

Dr. Melissa Magnuson has always wanted to be a veterinarian and fulfill her passion for helping animals.

She has a degree in biology and philosophy from Hamline University in St. Paul, MN, and a master’s degree from Southern Mississippi University. She was one of nine researchers to be trained to run an automated DNA sequencer, and helped map the pine genome. From there, she completed her DVM at the University of Minnesota, where she participated in mapping the pig and canine genomes. Dr. Magnuson has a special interest in surgery, emergency medicine, and avian and exotic animal care, and owns and operates three AAHA accredited small animal and exotic veterinary practices in New Hampshire. p.8

8. LISA MILLER, DVM, CCRT, CVA

Dr. Lisa Miller is VP of Clinical Veterinary Medicine for Companion Animal Health, and a graduate of the University of Tennessee, College of Veterinary Medicine. She is certified in canine rehabilitation therapy, and practiced rehab, sports medicine, and acupuncture before returning to general practice, and later joining the animal health industry. Dr. Miller has been working with laser therapy/photobiomodulation (PBM) since 2006. She coordinates all veterinary and advanced PBM research involving Companion Animal Health technologies, and is a consultant and reviewer for PBM research worldwide. She is a member of the AVMA, the American Association of Rehabilitation Veterinarians, and the American Society of Laser Medicine & Surgery. p.32

9. MARGO ROMAN, DVM, CVA, COT, CPT, FAAO

Dr. Margo Roman graduated from Tuskegee University Veterinary School in 1978, and did undergraduate studies at the University of Florida. She owns the integrative veterinary practice M.A.S.H. (Main Street Animal Services of Hopkinton in Massachusetts, www.mashvet.com). Dr. Roman has been using Medical Ozone Therapy for the past 20 years and Microbiome Restorative Therapy (MBRT) for 12 years, enhancing her 45 years of practicing both conventional and alternative modalities (e.g. acupuncture, homeopathy, herbs, nutrition, hyperbaric oxygen, UVBI). At home, Dr. Roman has seven standard poodles, sixth generation, and two Siamese cats, who have been donors for over 30,000 MBRT cases. p.44

editorial

DIGESTIVE HEALTH IS

FOUNDATIONAL HEALTH I have treated many gastroenterology cases, one of them a four-year-old female German Shepherd with a history of intermittent vomiting, reduced appetite, and lethargy. After further testing, she was diagnosed with inflammatory bowel disease. My treatment protocol involved switching her to an elimination diet, along with immunosuppressant medication to control inflammation. I also used probiotic supplementation. After a few weeks, the dog’s symptoms had resolved, illustrating the importance of accurate diagnoses, tailored treatment plans, and the use of probiotics for GI-based problems.

Gastrointestinal issues may be deemed the most common reason for veterinary visits. These disorders of the stomach and intestines can have a significant impact on an animalʼs quality of life. In this special gastroenterology-themed issue, we aim to explore digestive health and innovative treatment options from a variety of angles. For example, Iʼm glad we’re including a groundbreaking article by Dr. Margo Roman about microbiome restorative therapy (MBRT), which should reinforce the significance of microbiome health to readers. I always give importance to the microbiota and probiotic treatments, since a growing body of evidence indicates that the intestinal microbiome plays a role in the emergence, and subsequent maintenance, of a number of disorders. The makeup and functionality of the gut microbiota are influenced by a range of environmental variables. Keeping these variables in mind places us in a better position to understand the overall condition of the patient.

Successful outcomes begin with a thorough diagnosis.

Numerous laboratory tests for both dogs and cats can

help you reach a sound diagnosis, but the information you are able to extract and the clinical mapping you do is of pivotal importance — and it all can be achieved by asking the right questions. Malnutrition, obesity, adverse food reactions, diseases, nutritional therapy (including soluble fiber and probiotics), conventional treatment protocols, alternative therapies, and last but not least, integrative approaches, should all be taken into consideration for the successful diagnosis and treatment of gastroenterological problems.

Along with MBRT, we also feature other gastroenterology topics in this issue, including an integrative approach to chronic canine diarrhea by Dr. Ron Carsten, and TCVM treatment options for feline colitis by Dr. Chris Bessent.

In addition, you’ll find articles focusing on advances in veterinary diagnostics (Dr. Jean Dodds), and longevity in animals as seen through the lens of genetics (Dr. Melissa Magnuson).

This issue of IVC Journal offers a wealth of knowledge for veterinary professionals. I hope you enjoy it, and find the information helpful for your own practice.

Best regards,

Omer Rashid, DVM, MSc Parasitology Associate Editor, dromer@redstonemediagroup.com

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INDUCING LONGEVITY IN COMPANION ANIMALS:

WHAT DO GENETICS TELL US?

BY MELISSA L. MAGNUSON, DVM

The genetics of companion animals are affected by many factors, including diet, environment, and stress. By controlling these three aspects, we help care for our patients’ genes and extend their longevity.

There are two threats to well-being, as Dr. Deepak Chopra writes in Super Genes, a book he co-authored with Dr. Rudolph Tanzi. These threats are illness and aging, and they are constantly present. “Out of sight, without your knowing it, your present good health is being silently undermined,” Dr. Chopra writes.1 This is what we are forever up against as we try to “interrupt” the dying process in our animal patients. Every client wants their pet to live forever, or at least for as long as possible. No veterinarian has a magic pill to make this happen; if we did, we would be using it every day. I believe “our magic” lies in “caring” for the genes inside our patients. This genetic

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information holds answers to many of the questions we try to answer for our clients. What drives longevity in pets? And what can genetics tell us?

DOGS AS A MODEL FOR AGING Much has been studied about aging and scientists are currently using the dog as a model for aging in humans.

Why the dog? Researchers have noted that dogs have been on the same evolutionary path as humans, and since they live with us, they share the same environments and stressors

we do.2 Dogs also naturally develop age-related cognitive decline, with behavioral and histological characteristics that are very similar to those in humans. Researchers also know the genetic and biochemical pathways by which aging is controlled, and have identified nine common denominators.

3 These hallmarks are: genomic instability, telomere attrition, epigenetic alterations, loss of proteostasis, deregulated nutrient-sensing, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, and altered intercellular communication.3 Half these hallmarks focus on DNA and genetic pathways.

GENETIC INSTABILITY AND AGING Genetic injury or instability is one of the nine hallmarks of aging.3 I will focus only on genetic instability as this topic on its own is immense. The integrity and stability of DNA is continuously challenged by many exogenous and endogenous events. There are too many to detail, so for the purpose of this article we will focus on those we can control, including diet, the environment, and stress.

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DIET

Diet is not as simple as recommending just any bag of food on the shelf. Pet diets have become very important; consumers are educating themselves on

the best diets for themselves, and want that same level of nutritional quality for their pets. I surmise that some highlyeducated owners, thanks to access to peer-reviewed studies, have already done a great deal of research on their pets’ dietary needs.

There are so many options for pets as far as food is concerned.

The discussion in this arena is broad and controversial among many veterinarians. From my experience seeing birds and exotics, in addition to dogs and cats, replicating a diet in the wild is most beneficial, and keeping it in moderation is key.

Feeding one thing all the time is not ideal. When we do this, we create a microbiome that is lacking in diversity and susceptible to insult (stressors). I call these animals my “Sensitive Sallys”.

They’re the pets that fall ill if they eat anything besides one certain food; this tells me their microbiome is not diverse and the genetic variation has not evolved to handle stressors.

Another factor of pet diets is food quality. Processed foods have preservatives, dyes, synthetic additives and chemicals that act as stressors on the genetic material in the microbiome. Also, thanks to chemical farming, much of our food is devoid of nutrients before it is even further processed to make pet food. Though a few studies have been done in this area, I believe it’s something that requires more independent research.

Microbiome testing can be beneficial for

GOOD DNA ISN’T THE WHOLE PICTURE You may think that if a dog is born with good genes, they will live a long time. While good DNA is “part” of it, it certainly isn’t the whole story. What is actually more important is taking care of the genes inside the dog, and that the dog has a good mechanism for repairing DNA when it becomes injured. Then they can live a long time.

This includes not just the genes inside the 20 trillion cells that make up the dog, but also the genetic material in their microbiome4 -the millions and millions of bacteria that are coding and replicating on a daily basis. When genes become damaged, whether inside the cell or the microbiome bacteria, mutations occur. Mutation allows for genetic variability, and most of the time causes evolution.

But sometimes it has deleterious effects and ends in disease or even death. The bigger question is what damages these genes, why don’t they get repaired, and what can we do about it?

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understanding how a pet’s microbiome is currently working, and allows us to help regulate bacteria and general overall health. Some companies have done much work in this area to help our dogs’ and cats’ microbiomes build themselves with more variation and stability.

In my experience (based on 25 years of vetting pets), keeping animals lean is vital to longevity. Two key factors keep our pets lean — the food they eat and the activity they receive. Balancing these two factors keeps the animal lean — not average or a little heavy, but a little less than average, with a BSC of 3 or 4 out of 9). Research on longevity in nematodes, yeast, fruit flies, mice and rhesus monkeys shows that dietary restriction causes specific nutrient sensitive pathways to become more protective and conserve DNA.5 Though the following is a study of one, I had a Chocolate Labrador Retriever with severe heart disease live to be 17 years of age. He was very lean when diagnosed with heart disease and the owner was compliant with medications and veterinary visits. She focused on diet, exercise, medications for the heart, and lots of TLC. Typically, heart disease in dogs does not parallel longevity. Most pets (and humans) with heart disease are overweight. In this Labrador’s case, weight was eliminated as an underlying factor, and he flourished in his geriatric years.

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ENVIRONMENT

A pet’s environment can be managed by providing appropriate shelter, exercise that replicates being “in the wild”, and paying attention to the effects of sun or lack thereof. Vitamin D levels have not been studied extensively in pets, except when it has been over-supplemented.6 Pet foods are fortified with vitamin D since many of animals do not go outside. Feeding commercially-prepared diets (when fed correctly) does provide the proper amount of vitamin D; however, with the plethora of diets

available, owners home-cooking and supplementing on their own, vitamin D screening becomes, I believe, a crucial diagnostic. Human studies have shown that as people age, vitamin D levels decrease and supplementation is necessary.

Vitamin D is necessary for gene expression and has a wide variety of functions in more than 160 pathways linked to cancer, autoimmune disorders, and cardiovascular disease.7 From reading the studies, this area may be useful in understanding how nutrients and nutrition affect a pet’s genomic expression.

Other environmental factors affecting gene expression include air quality, emotional stability within the home, and the time the owner has for the pet. Our pets are social creatures, and it is imperative we give them time so they are emotionally as well as nutritionally cared for. Human studies are not abundant, but researchers are starting to look very closely at DNA methylation and the maltreatment of children at a young age.8 This is a complex topic, but there are theories that biological genetic development may be influenced more than we think by psychological environmental factors.

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STRESS

Stressors are abundant, and can be anything that causes an imbalance to homeostasis. Diet and environment are discussed above, but things like chronic pain, chronic inflammation, or chronic disease can affect pets at a cellular level. We often think of cancer when we consider chronic inflammation and disease. Genetics helps us understand cancer evolution a bit better. The

WHY UNDERSTANDING GENES IS SO IMPORTANT When I was sequencing the pig and canine genome back in 1994, I could never have fathomed the impact that understanding genes would have on unravelling how life really works.

Genes hold DNA, the sophisticated and complex molecule that contains all the information for growing and sustaining any organism.

DNA is handed down to offspring, sometimes as an exact copy and sometimes rearranged to become better, worse, or just different.

Knowing and understanding how DNA works, what affects it, and how it replicates has become an enormous area of study. Because this hereditary information is passed along to many generations, we are able to see the vast impact that diet, environment, and stress have on genetic information.

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problem with cancer is not that cell function is weakened by damage, but that it’s too vigorous.5 Most disease etiology is a combination of molecular (DNA) damage and hyperfunction.5 Understanding hyperfunction is best explained by considering a young pet that we feed to help with growth and development. When the animal becomes an adult, we no longer need to support growth and development; but by overfeeding the pet (in the exam room I call it “over-loving”), we cause the genetic and cellular pathways to work at the same level they did during the growth and developmen phase of life.

Hypertrophy and hyperplasia underlie many diseases, including cardiovascular, diabetes, cancer, inflammation, adipose and hyperglycemia.5 The progression of cancer is more a problem of too much than too little. Overworking our systems with too much could actually be the very problem causing our DNA to become damaged. Eliminating hyperfunction is protective.

Damage to cells from stressors, whether it be endogenous or exogenous, causes DNA to be injured and in need of repair.

These repair mechanisms (there are many) are vital to keeping DNA in check.9 When repair mechanisms are no longer doing

their job, they can cause accelerated aging and disease. Much research is being done on DNA repair, and the reasons why repair mechanisms sometimes work and sometimes don’t is still not fully understood.9 When they do not work, we feel like we are dead in the water. Disease happens, cancer happens, our pets age faster than we thought, and turning it around can seem an insurmountable task.

Focusing on prevention is much easier than trying to fix a broken genetic or cellular pathway after years of it being bombarded by the infinite array of DNA damaging agents.

Working toward a chemical-free, preservative-free, organic, emotionally stable, and connected world, where we focus on the basics of care, clean and nutritious food, and optimal husbandry, is the best place to start. Preventative care, instead of reactionary care, is the key to longevity in our pets. _____________________________________________________ Deepak Chopra MD, Rudolph E. Tanzi PhD, Super Genes, Crown Publishing 2015.

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Sara Sandor and Eniko Kubinyi, Genetic Pathways of Aging and Their Relevance in the Dog as a Natural

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Model of Human Aging, Front Genet. 2019; 10: 948. Published online 2019 Oct 18. doi: 10.3389/ fgene.2019.00948.

Carlos Lopez-Otin, Maria A. Blasco, Linda Partridge, Manuel Serrano, Guido Kroemer, The Hallmarks of

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Aging, Cell. Author manuscript; available in PMC 2013 Nov 21. Published in final edited form as: doi: 10.1016/j.cell.2013.05.039.

Jan S. Suchodolski, Anaysis of the gut microbiome in dogs and cats. Vet Clin Pathol. 2022 Feb;

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50(Suppl 1): 6–17. Published online 2021 Sep12. doi: 10.1111/vcp.13031.

David Gems, Linda Partridge, Genetics of Longevity in Model Organisms: Debates and

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Paradigm Shifts, Annu. Rev. Physiol. 2013. 75:621–44 First published online as a Review in Advance on November 26, 2012. The Annual Review of Physiology is online at http://physiol.annualreviews.org This article’s doi: 10.1146/annurevphysiol-030212-183712.

HAW Hazewinkel, MA Tryfonidou, Vitamin D3 metabolism in dogs, Mol Cell

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Endocrinol, 2002 Nov 29;197(1-2):23-33. PMID: 12431792, DOI: 10.1016/ s0303-7207(02)00275-7.

J. Christopher Gallagher MD MRCP, Vitamin D and Aging, Endocrinol Metab

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Clin North Am. Author manuscript; available in PMC 2014 Jun 1. Published in final edited form as: Endocrinol Metab Clin North Am. 2013 Jun; 42(2): 319–332. Published online 2013 Apr 9. doi: 10.1016/j.ecl.2013.02.004.

Stephanie H Parade, Lindsay Huffhines, Teresa E Daniels, Laura R Stroud, Nicole

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R Nugent, Audrey R Tyrka, A systematic review of childhood maltreatment and DNA methylation: candidate gene and epigenome-wide approaches, Transl Psychiatry. 2021; 11: 134. Published online 2021 Feb 19. doi: 10.1038/s41398021-01207-y.

Nimrat Chatterjee, Graham C Walker, Environ Mol Mutagen. Author manuscript; available in

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PMC 2018 Jun 1. Published in final edited form as: Environ Mol Mutagen. 2017 Jun; 58(5): 235–263. Published online 2017 May 9. doi: 10.1002/em.22087.

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HOW VETERINARY DIAGNOSTICS HAVE ADVANCED OVER THE LAST 50 YEARS

BY W. JEAN DODDS, DVM

Thanks to a range of scientific and clinical advancements, veterinary diagnostics, along with disease management and treatment options, have come a long way since the 1970s.

The development and application of clinical innovations in both human medical and veterinary diagnostics have evolved dramatically over the last five decades.1-7 Today, diagnostic, management, and treatment options are improved even further by focusing on the individual patient, as in personalized medicine for people and companion animals.3,8 Additional factors, including One Health (see sidebar on page 33) and significant advancements in the field of genetics, have further improved diagnostics for both human and veterinary patients.

VETERINARY DIAGNOSTICS

BY GENETICS, ORGAN AND FUNCTION It may be surprising to readers that there is so much more literature on veterinary diagnostics than there is for humans. It becomes more understandable when we recognize that animals often serve as models before any findings are adapted to humans.8 While an equal number of studies have been done in livestock and wildlife,15 this review primarily covers a selected group of studies in companion animals.

• Behavioral disorders and the brain: Stress, whether emotional, physical, chemical or metabolic, plays a major role in behavior as measured by biomarkers of cellular

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oxidative stress.9,17,18,19 Recent studies have shown that cannabinoids (CBD) help control stress not only in people but also in veterinary patients. Over a six-month study period, a once daily dose of broad-spectrum THC-free CBD was shown to be efficacious and safe in relieving stress anxiety from being left alone or during car travel.20 Additionally, the importance of the gut-brain axis in regulating the microbiome and gut health has resulted in improved diagnostics, management, and therapy.9,19,21-24 Several commercial entities now offer microbiome analyses for pet animals.24

• B leeding disorders: A recent major breakthrough in gene therapies for humans with hemophilia A (factor VIII deficiency) and B (factor IX deficiency), has led to effective and safe cures for these disorders, albeit at a very high cost in the millions of dollars.25,26 The commercial products are Roctavian (valoctocogene roxaparvovec) for hemophilia A,25 and Hemgenix (etranacogene dezaparvovec) for hemophilia B.26 Hopefully, these research and therapeutic advances will soon be applied to parallel companion animal hemophilias, although obtaining the needed funding will be challenging.

In beagles with delayed post-operative hemorrhage (DPOH), a missense G96E mutation encoding for alpha-2 antiplasmin in the F7 gene was found at exon 5, with

glycine26 substituted by glutamic acid136; this mutation has a frequency of 31% in the breed.12 The effect of this genetic variant on bleeding susceptibility was reported as mild to moderate.8,12 More recently, another missense variant mutation in the SERPINEF2 gene encoding alpha-2 antiplasmin was identified by GWAS on chromosome 9, with a relatively high prevalence in Scottish deerhounds that were also affected by DPOH.10 Platelet lysates for human cell therapy not only provide for improved hemostasis but also contribute to regenerative medicine by helping control inflammation and infection, and supporting wound healing.27 Parallel studies with platelet lysates are ongoing in companion animals, including horses.28, 29

• Cancers: Diagnostics for the early detection of cancers in humans and animals, in addition to the conventional techniques of pathology, biopsy and histology, now include nanotechnology, genetic copy number variations, cytological grading systems, specialized ultrasound, cellular and serum zbiomarkers, flow cytometry, stem cell technologies, patterns of lymphocytic infiltrates, immunohistochemical expression, expression of sex hormone receptors, and liquid biopsies.1,2,5,6,9,14,30-33

• C ardiac disorders: Controversial issues surrounding the diagnosis and causes of canine heart disease, specifically dilated canine cardiomyopathy, have escalated to include genetic predisposition, dietary components, environmental inhalant and contact exposures, vaccines, toxins, parasiticide preventives and others.9,21,22

• C RISPR applications: The decades-long genomic revolution culminated in the recent Nobel Prize-winning CRISPR Cas9 and Cas11-13 research and its ensuing patent disputes between the University of California, Harvard University and MIT’s Broad Institute.8,34 This has since evolved to include novel genome editing tools such as Prime and PASTE.34

• Endocrine and metabolic disorders: Endocrine disruption, deficiencies, and excess production have been recognized for many years, especially when involving the diet and microbiome, and pituitary, thyroid, pancreas, adrenal, and parathyroid glands.9,35-38 Studies of the pituitary-thyroid and adrenal axes, and the novel reference laboratory in-clinic/

THE IMPACT OF

ONE HEALTH ON DIAGNOSTICS The concept of One Health, where all sentient beings learn from and benefit each other, has provided a strong impetus for exploration and discovery.8 For example, take a look at the scientific history of bleeding disorders:8 • 1 700s — The process of blood clotting was linked to hemostasis (the cessation of bleeding).

• 1 800s — thrombosis was first recognized by Virchow; platelets were discovered by Bizzozero; familial bleeding tendency in males (hemophilia) was first recognized.

• 1 900s — Morawitz described the classical theory of blood coagulation.

• 1 930s-1940s — disputes arose between scientists about the factors that form and dissolve clots; more clotting factor disorders were recognized in people (von Willebrand Disease, vWD) and dogs (hemophilia).

• 1 950s — vWD was identified in Poland-China pigs; factor VII was identified in dog plasma after coumarin therapy prolonged blood clotting in vitro.

• 1 970s-1980s — the evolution of human and veterinary clinical pathology/diagnostic medicine began with early hand-held plain and silicone-coated test tubes dipped in and out of a water bath kept at body temperature; and has evolved to sophisticated nanotechnology and molecular arrays and other plasma biomarker assays. vWD was described in German Shepherd Dogs imported to North America from Germany, and then in many dog breeds, cats, and rabbits; hemophilia was described in cats and horses; factors I, IX, X, XI and XII deficiencies were documented in dogs, cats, cattle, goats; and platelet defects were described in dogs, rats, and mice.

• 1 990s-today — more of these bleeding disorders were found in domestic and companion animals, including familial pre-kallikrein and kallikrein deficiencies. Gene editing and therapy was developed and applied to several heritable human coagulopathies.

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bedside diagnostic testing of humans and companion animals,have made diagnosis more accurate along with emphasizing the need for species-specific reagents and dynamic metabolic testing.3,7,9,35-38 Parallel concerns apply to the increasing number of metabolic disorders, with emphasis on expanding global numbers of inflammatory conditions; autoimmune disorders, especially diabetes, thyroiditis, and Addison’s disease; infectious and antibiotic-resistant microbial diseases; increasing gluten sensitivities; and the widespread issues arising from obesity in people and pet animals.9,39,40

• Gastrointestinal disorders: As the critical importance of the microbiome in balancing the gut-brain and gut-lung axes has evolved, new diagnostic and therapeutic approaches have become available, including stem cell infusion and fecal transplantation (microbial restoration therapy) from healthy, matched donors.9,21,22,24,38-43 Commercial availability now extends to animal patients.9,21,39,41,42

• Genetic disorders: Many of the conditions discussed above occur more often in people and animals carrying a specific heritable trait or a predisposition to them.8-14,25,26,34-37,39

GENOME SEQUENCING HAS CHANGED MEDICINE Genome sequencing has now been completed for humans and 14 other animal species (primates, rodents, farm animals, and companion animals).7-14 The application of genome wide association sequencing (GWAS), and gene editing with CRISPR and other emerging technologies and therapies, has changed the face of scientific and clinical medicine by enhancing the health, well-being, and longevity of all living things.1,8,9,13,15 However, not all genetically based diagnostic tests are accurate and reliable, as recently documented for canine DNA testing.16

• Hematologic disorders: In addition to the bleeding disorders already discussed, other human and animal blood disorders have been documented.9,44,45 Companion animal blood typing is performed for the purposes of identifying the transfusion needs of patients and blood donors; establishing techniques in regenerative and stem cell medicine; and determining potential mismatched blood type mating that can lead to hemolytic disease of newborn dogs, cats, horses, and livestock.9,44-47

around time for completion, and their increased cost.

1-7,43,52-57 Adding to this challenge is the ever-increasing development of microbial strains becoming resistant to standard antimicrobials, which now require novel, more costly therapies with limited availability that need to be given for extended periods in both people and animals.1,3,4-7,43,51,52, 56, 57

• Hepatic disorders: As a major organ for the production of coagulation proteins and hematopoietic cell storage, as well as reticuloendothelial cell function and clearance, the liver plays a pivotal role in health and disease conditions.8,9 Similar physiological and pathological functions apply to humans and other species.48-51

• Infectious diseases and prevalence: Microbial infections from bacteria, viruses, fungi (including yeast), protozoa, phages, and parasites are much more prevalent than the other conditions discussed above. However, the worldwide success rate for early and accurate identification is hampered by factors such as the varying ability of even the most accurate tests to predict clinical results, their turn

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Regarding the prevention of infectious diseases, vaccines play a preeminent role but have been plagued by controversy ever since the SARS-CoV-2 pandemic of COVID-19 and the advent of life-saving mRNA vaccine technology.57 Similar vaccine hesitancy and naysayers are vocal in veterinary medicine, such that refusal to vaccinate has led to the inevitable increased risk of vaccine-preventable infectious diseases, suffering and even death.58,59 Issues regarding vaccine safety and adverse events include injection site sarcomas, adjuvant-induced autoimmune reactions, thimervosal (mercury) and aluminum heavy metal content, and squalene content, as well as the immunizing volume of vaccine needed for very small versus larger pets.58,59

• Renal and urinary tract disorders: Like the liver, the kidneys detoxify the body of small proteins, drugs, and fluids in relation to the glomerular filtration rate. Congenital abnormalities and acquired disorders affect kidney size, number, anatomy (hypoplasia, dysplasia, polycystic), and function in humans and animals.9 At least 14 dog breeds have heritable, familial renal disease or are genetically predisposed, ranging from the small Shih Tzu to the much larger Doberman Pinscher, Akita, and Alaskan Malemute, where the renal disease is associated with dwarfism.60 Disorders of the urethra, ureters (e.g. ectopic ureter especially in female pups), and bladder also occur.60

Improved diagnostics that help us find disease processes faster and earlier often lead to better outcomes for our animal patients.

The scientific advances of the last 50 years have greatly enriched our ability to get to the bottom of our patients’ health problems, so we can implement a treatment or management plan as soon as possible.

1Sarkar AA. Biotech Horizon. Molecular diagnostics adapts to evolving diseases. Gen Eng Biotech News

2023, 43 (2); 40, 49, 50. www.GENengnews.com.

2 Stein RA. The many facets of flow cytometry. Gen Eng Biotech News 2023, 43 (2); 44-47. www.

GENengnews.com.

3Soreng K. Advances in personalized medicine: prognostic testing in patient management.

Med Lab Observer 2023, 55 (3); 18-21. www.MLO-Online.com.

• Respiratory disorders: Interest in human respiratory disorders, as exemplified over the years by infections like bacterial and parasitic pneumonia, tuberculosis, and the common cold, has grown exponentially with the advent of the COVID-19 pandemic, and co-infections with seasonal human influenza virus, rhinoviruses, enteroviruses, and respiratory syncytial virus.1,57,61 In animals, similar issues and controversy have arisen with the zoonotic transmission of these novel coronavirus variants from people to wildlife and pets, and vice versa.59 Other respiratory infections in animals include canine parainfluenza virus 5, with improved diagnostic identification.43,54 The current global spread of virulent avian influenza (bird flu) H5N1-N8 strains from migrating wild birds has devastated commercial and backyard poultry flocks. Certain mammals like the red fox are also susceptible,62 although it very rarely causes respiratory infection in humans.62 The importance of the gut-lung axis and the role of the microbiome have also been explored with regard to respiratory disorders.24,56

4Frew E.

For emerging infectious diseases, reliable controls support QC efforts. Med Lab Observer 2023, 55 (3); 14-16. www.MLO-Online.com.

5Clifford L-J. Digital pathology and clinical testing: considerations for a successful integration. Med Lab

Observer 2023, 55 (3); 22-23. www.MLO-Online.com.

6Oelmueller U, Safwat N. Liquid biopsies: brilliant potential but highly workflow dependent.

Med Lab Observer 2023, 55 (3); 34, 36. www.MLO-Online.com.

7Nadeau K. State of the Industry. Laboratory data analytics. Med Lab Observer 2023, 55 (3); 24-26, 28,

30, 32. www.MLO-Online.com.

8 Dodds WJ. One Health: Animal models of heritable human bleeding diseases. Animals 2023, 13; 87.

doi.org/10.3390/ani3010087.

9 Dodds WJ. Scientific Reports: Epigenetics. Programming for health and longevity. J Am Hol Vet Med

Assoc 2014, 37;16-22.

10 Court MH, Kiser JN, Neibergs HL, Zhu Z, Dillberger JE. Identification by whole genome sequencing

of genes associated with delayed postoperative hemorrhage in Scottish deerhounds. J Vet Intern Med 2023; 1-8. doi:10.1111/jvim.16643.

11Parker HG, Dreger DL, Rimbault M, et al. Genomic analyses reveal the influence of geographic origin,

migration, and hybridization on modern dog breed development. Cell Rep 2017,19;697-708.

12Callan MB, Aljamali MN, Margaritis P, et al. A novel missense mutation responsible for factor VII

deficiency in research beagle colonies. J Thromb Haemost 2006, 4; 2616-2622.

13Donner J, Anderson H, Davison S, et al. Frequency and distribution of 152 genetic disease variants

in over 100,000 mixed breed and purebred dogs. PLoS Genet 2018, 14(4): e1007361. doi. org/10.1371/ journal.pgen.1007361.

14 Aupperle-Lellbach H, Heidrich D, Kehl A, et al. KITLG copy number germline variations in schnauzer

breeds and their relevance in digital squamous cell carcinoma in black giant schnauzers. Vet Sci 2023, 10; 147. doi.org/10.3390/vetsci10020147.

15Scocco P, De Felice E, Malfatti A. Immunohistochemical and physiological research on farm animals.

Animals 2023, 13; 739. doi.org/10.3390/ani13040739.

39 Kolb N, Alf V, Cappello R, et al. Novel association of gluten sensitivity with immune-mediated

neuromuscular syndrome in a dog. Neuromuscular Disorders 2022, 32; 903–907.

40 Corda F, Ballocco I, Corda A, et al. Coagulation abnormalities in dogs with parvoviral enteritis. Vet Sci

2023, 10 (1), 41. doi.org/10.3390/vetsci10010041.

41Higueras C, Escudero R, Rebolé A, er al. Changes in faecal and plasma amino acid profile in dogs with

16CBC News Marketplace. How accurate are dog DNA tests? CBC News 2023, March 4, 2023 www cbc.

ca/news/business/marketplace-dog-dna-test-1.6763274.

food-responsive enteropathy as indicators of gut homeostasis disruption: a pilot study. Vet Sci 2023, 10; 112. doi.org/ 10.3390/vetsci1002011.

17 Guelfi G, Capaccia C, Santoro MM, et al. Identification of appropriate endogenous controls for

42El-Wahab AA, Lingens JB, Hankel J, et al. Effect of different fiber sources as additives to wet food for

circulating miRNA quantification in working dogs under physiological stress conditions. Animals 2023, 13; 576. doi.org/10.3390/ani13040576.

beagle dogs on diet acceptance, digestibility, and fecal quality. Vet Sci 2023, 10; 91. doi.org/10.3390/ vetsci10020091.

18 Ogi A, Gazzano A. Editorial: Biomarkers of stress in companion animals. Animals 2023, 13(4), 660.

43Xiao X, Xiangqi Hao X, Bo Chen B, et al. Two multiplex PCR methods for detecting several patho-

doi: 10.3390/ani13040660.

19 NHGRI. Autism susceptibility in mouse models linked to endogenous retroviruses. Gen Eng Biotech

News 2023, 43 (4); March 13, 2023. www.GENengnews.com.

20 Hunt ABG, Flint HE, Logan DW, et al. A single dose of cannabidiol (CBD) positively influences

measures of stress in dogs during separation and car travel. Front Vet Sci 2023, 10; 11126044, Feb 22, 2023. doi:10.3389/fvets.2023.1112604.

21Dodds WJ. Diagnosis and management of adverse food reactions. Biomed J Sci &Tech Res 2018, 3(2).

doi:10.26717/BJSTR.2018.03.000868.

22Dodds WJ. Diagnosis of canine food sensitivity and intolerance using saliva: report of outcomes. J

Am Hol Vet Med Assoc 2017/2018, 49: 32-43.

23Balouei F, Stefanon B, Sgorlon, et al. Review. Factors affecting gut microbiota of puppies from birth

to weaning. Animals 2023, 13; 578. doi.org/10.3390/ani13040578.

gens associated with feline respiratory and intestinal tracts. Vet Sci 2023, 10 (1), 14. doi.org/10.3390/ vetsci10010014.

44 Giger U, Idalan N, Caviezel L, Raj K. Controversies and updates on immunodiagnostics of IMHA in

dogs. Proc of ACVIM Forum, 2021. www.vin.com.

45Miglio A, Valente C, Guglielmini C. Red blood cell distribution width as a novel parameter in

canine disorders: Literature review and future prospective. Animals 2023, 13, 985. doi.org/10.3390/ ani13060985.

46Véran E, Blais M-C. Validation of a cage-side agglutination card for Dal blood typing in dogs. J Vet

Intern Med 2023, March 2. doi:10.1111/jvim.16646.

47Kaneps AJ. A one-health perspective: use of hemoderivative regenerative therapies in canine and

equine patients. J Am Vet Med Assoc 2022, 261 (3); 301-308. doi.org/10.2460/javma.22.12.0556.

48 Balkman CE, Center SA, Randolph JF, et al. Evaluation of urine sulfated and nonsulfated bile acids as

24Dodds WJ. Your pet’s microbiome. Animal Wellness 2022, 24 (3); March 24, 2022.

a diagnostic test for liver disease in dogs. J Am Vet Med Assoc 2003, 222 (10); 1368-1375. doi:10.2460/ javma.2003.222.1368.

25Mahlangu J, Kaczmarek R, von Drygalski A, et al. Two-year outcome of valoctocogene roxaparvovec

49 Pena-Ramos J, Barker L, Saiz R, et al. Resting and postprandial serum bile acid concentrations in

therapy for hemophilia A. N Eng J Med 2023, 388; 694-705. doi:10.1056/NEJMoa2211075.

dogs with liver disease. J Vet Intern Med 2021, 35;1333–1341.

26Pipe SW, Leebeek FWG, Recht M, et al.

50 Phillips RK, Steiner JM, Suchodolski JS, et al. Urinary 15-F2t-isoprostane concentrations in dogs with

Gene therapy with etranacogene dezaparvovec for hemophilia B. N Eng J Med 2023, 388; 706-718. doi:10.1056/NEJMoa2211644.

27Delabie W, De Bleser D, Vandewalle V, et al. Single step method for high yield human platelet lysate

production. Transfusion 2023, 63; 373-383. doi:10.1111/trf.17188.

28 Goggs R, Brainard BM, LeVine DN, et al. Lyophilized platelets vs cryopreserved platelets for

management of bleeding in thrombocytopenic dogs: a multicenter randomized clinical trial. J Vet Intern Med 2020, 34; 2384-2397. doi:10.1111/jvim.159330.

29 Hagen A, Holland H, Brandt VP, et al. Platelet lysate for mesenchymal stromal cell culture in the

canine and equine species: analogous but not the same. Animals 2022,12(2):189. doi: 10.3390/ ani12020189.

30 Labant MA. Taking the measure of cancer genomically. Gen Eng Biotech News 2023, 43 (4); 24-28.

liver disease. Vet Sci 2023, 10; 82. doi.org/10.3390/ vetsci10020082.

51Konstantinidis AO, Patsikas MN, Papazoglou LG, et al. Congenital portosystemic shunts in dogs and

cats: classification, pathophysiology, clinical presentation and diagnosis. Vet Sci 2023, 10; 160. doi. org/10.3390/vetsci10020160.

52Dodds WJ. An update on vaccines and titer testing for dogs and cats. Animal Wellness 2019,

21(5);50-52.

53Sweet A, Andre N, Licitra BN, et al. RNA in-situ hybridization for pathology-based diagnosis of feline

infectious peritonitis (FIP): current diagnostics for FIP and comparison to the current gold standard.

Qeios 2022, June 22. doi.org/10.32388/NUN8KB.3.

54Jeon G-T, Kim H-R, Shin Y-K, et al. An improved duplex real-time quantitative RT-PCR assay with

www.GENengnews.com

a canine endogenous internal positive control for more sensitive and reliable detection of canine parainfluenza virus 5. Vet Sci 2023, 10;142. doi.org/10.3390/ vetsci10020142.

31Kaszak I, Witkowska-Piłaszewicz O, Domrazek K, et al. The novel diagnostic techniques and bio-

55Schäfer I, Kohn B, Silaghi C, et al. Molecular and serological detection of Anaplasma phagocytophilum

32Dilley KN, Wong A, Kent MS, et al. Expression of sex hormone receptors in canine osteosarcoma. Vet

56Morgan J. Biofilms and how they affect your dog and cat. Animal Wellness 2023, 25(2);56-58.

markers of canine mammary tumors. Vet Sci 2022, 9; 526. doi.org/10.3390/vetsci9100526.

Sci 2022, 9; 524. doi.org/10.3390/vetsci9100524.

in dogs from Germany (2008–2020). Animals 2023, 13; 720. doi.org/10.3390/ani13040720.

squamous cell carcinoma in dogs. Vet Sci 2023, 10; 86. doi.org/10.3390/vetsci10020086.

Comparing COVID-19 vaccines for their characteristics, efficacy and effectiveness against SARS-CoV-2 and variants of concern: a narrative review. Clin Microbiol Infect 2022, 28(2):202-221. doi:10.1016/j.cmi.2021.10.005.

34Philippidis A. PASTE, don’t cut: genome editing tool looks beyond CRISPR and Prime. Gen Eng

58 Dodds WJ. Review Article. Vaccine issues and World Small Animal Veterinary Association guidelines

33Monteiro A, Delgado L, Monteiro L, et al. Immunohistochemical expression of Tensin-4/CTEN in

Biotech News 2023, 43 (4); March 10, 2023. www.GENengnews.com.

35Dodds WJ, Laverdure DR. The Canine Thyroid Epidemic, 2011; 192 pp. DogWise Publishing,

Wenatchee, WA.

36Dodds WJ. Diagnosing, managing and treating thyroid diseases in dogs and cats. IVC J 2022,

12 (4); Fall, 26-30.

37Dodds WJ. Guest Editor’s introduction to endocrinology. J Am Hol Vet Med Assoc 2015, 40; 8-21.

38 Buishand, F.O. Current trends in diagnosis, treatment and prognosis of canine insulinoma. Vet Sci

2022, 9; 540. doi.org/ 10.3390/vetsci9100540.

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57Fiolet T, Kherabi Y, MacDonald CJ, et al.

(2015-2017). Israel J Vet Med 2018, 73(2); 3-10.

59 Lyoo K-S, Yeo Y-H, Lee S-G, et al. Susceptibility to SARS-CoV-2 and MERS-CoV in beagle dogs.

Animals 2023, 13 (4); 624. doi.org/10.3390/ani13040624.

60 Fitzgerald SD. Congenital and inherited disorders of the urinary system in dogs. Merck Vet Manual

2022, October 2022. www.merckvetmanual.com.

61Chandra R. Diagnosing upper and lower respiratory tract infections. Med Lab Observer 2023, 55(2);

8-15. www.MLO-Online.com.

62CDC. HFN1 bird flu current situation summary. Centers for Disease Control and Prevention 2023, March

13. www.cdc.gov/flu/avianflu/avian-flu-summary.htm.

With all this in mind, probiotics no doubt already play an important role in your patient care for pets with gastrointestinal issues. But not all probiotics are created equal.

CHOOSING THE RIGHT PROBIOTIC FOR PET PATIENTS Finding a stable probiotic is key to ensuring that their beneficial organisms will survive the storage process and harsh stomach acids to do their healing work in the lower intestine.

SACCHAROMYCES BOULARDII A non-pathogenic strain of yeast, S. boulardii is an ideal probiotic because it can withstand the stresses of gastrointestinal transit and is resistant to all known antibacterial antibiotics.4 Randomized, placebo-controlled, doubleblinded studies have been reproduced to claim support from meta-analysis reviews that S. boulardii is an effective biotherapeutic agent for the treatment of antibiotic associated diarrhea, C. difficile-associated disease, and other causes of acute diarrhea in laboratory, human, and veterinary settings.5

GLUTAMINE Research has shown the amino acid glutamine to be a key intestinal nutrient for maintaining a healthy bowel environment.

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Digestive issues in dogs account for 10% of all veterinary visits. As a trusted veterinarian, you have surely seen your fair share of dog digestive woes — and know how debilitating they can be. You also know that a healthy bowel terrain is paramount to optimal gastrointestinal tract function.

More than this, it is the foundation of immune system competence and plays a pivotal role in the balance of neurotransmitters supporting mental health.

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BY DR KEITH WEINGARDT, DVM, CVA, CTCVM

Both strains can help reduce inflammation in the intestines by increasing butyric acid, other short chain-fatty acids, and beneficial enzymes,2 and enhance nutrient digestibility by reducing inflammation.3

Prebiotic dietary fibers are included in ThorneVetʼs Probiotic Support Formula, and act as carbon sources for fermentation pathways in the colon, supporting digestive health in multiple ways.

RI

A better understanding of probiotics, and the science backing specific probiotic ingredients, can help you harness the full power of their gut-healing potential for your patients.

B. coagulans and B. subtilis are stable, spore-forming, soil-based probiotics that are resistant to gastric acid, and thus able to pass through the stomach and enter the intestine in a similar state to their initial concentration.1

TO ER

To this end, ThorneVet has created a specially formulated, broad-spectrum pet Probiotic Support Formula, containing specific, proven-effective, gut-healing ingredients.

V AD

PREBIOTICS WHY BROAD SPECTRUM PROBIOTICS BACILLUS COAGULANS BACILLUS ARE BETTER AND SUBTILIS FOR PETS

The health benefits of inulin — also found in foods like artichokes, chicory and dandelion — range from the production of beneficial metabolites to a reduction in pathogenic bacterial populations, as well as improved immune system function.7 By providing comprehensive support for a healthy bowel terrain in the form of science-backed, specially formulated pet probiotics, we can help our patients live healthier, happier lives.

Dr. Keith Weingardt, a graduate of Cornell University College of Veterinary Medicine, International Veterinary Acupuncture Society, and Chi Institute of Traditional Chinese Veterinary Medicine, has started successful integrative veterinary practices in San Diego, CA and Portland, OR. A dedicated herbalist, he currently consults for ThorneVet, specializing in product development and continuing education.

Spinosa MR, Braccini T, et al. On the fate of ingested Bacillus spores. Res Microbiol 2000; 151: 361-368.

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Vinolo MAR, Rodrigues HG, et al. Regulation of inflammation by shortchain fatty acids. Nutrients 2011;3:858-876. doi: 10.3390/nu3100858.

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Acuff HL, Aldrich CG. Evaluation of graded levels of Bacillus coagulans GBI-30, 6086 on apparent nutrient digestibility, stool quality, and intestinal health indicators in healthy adult dogs. J Anim Sci 2021 May 1;99(5):skab137. doi: 10.1093/jas/skab137. PMID: 33940614; PMCID: PMC8158428.

3

B uts JP. Mechanisms of action of biotherapeutic agents. In: Elmer GW, McFarland LV, Suawicz CM, eds. Biotherapeutic agents and infectious diseases. Totowa, NJ: Humana Press, 1999;27-46.

4

Buts JP. Twenty-five years of research on Saccharomyces boulardii trophic effects: updates and perspectives. Dig Dis Sci 2009 Jan;54(1):15-18.

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Kim MH, Kim H. The roles of glutamine in the Intestine and Its Implication in Intestinal Diseases. Int J Mol Sci 2017 May 12;18(5):1051. doi: 11.3390/ijms18051051. PMID: 28498331; PMCID: PMC5454963.

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C arlson JL, Erickson JM, Lloyd BB, Slavin JL. Health effects and sources of prebiotic dietary fiber.

Curr Dev Nutr 2018 Jan 29;2(3):nzy005. doi: 13.1093/cdn/nzy005. PMID: 30019028; PMCID: PMC6041804.

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Glutamine suppresses pro-inflammatory signaling pathways and protects cells against cellular stresses during normal and pathologic conditions.6 IVC Summer 2023

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BY RON CARSTEN, DVM, PHD, CVA, CCRT

An e v i at r g e t n I h c a C o h r r o p n p i c A to

Diarrhea

Chronic diarrhea in dogs can have causes that range from something simple to a complex problem that is difficult to resolve. Optimal support for the latter patients relies on an integrative treatment plan that encompasses the interrelated factors contributing to this often challenging health problem.

Chronic diarrhea can be distressing for both dogs and their caregivers, especially when ongoing episodes occur in the house or at night. To add to the distress, dogs with chronic diarrhea (CD) often experience vomiting, appetite and weight loss, and pruritus. Not surprisingly, moderate to severe CD has been associated with reduced quality of life in dogs.1 Unfortunately CD has a long list of causes and is not specific for diseases of the digestive tract. This means a complete understanding of the potential triggers, diagnostic approaches, and management options is essential. Managing CD in a dog can be as simple as administering an appropriate deworming treatment — or as complex as diagnosing and treating chronic inflammatory enteropathy (CIE). Taking an integrative approach to chronic diarrhea in dogs can be an effective way to deal with this common and often debilitating problem.

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in Dogs

DEFINITIONS OF DIARRHEA VARY Generally, diarrhea is recognized as feces with abnormal consistency and excess water content. Stools can range from soft to liquid, without form. Diarrhea that has been present for three weeks or longer is considered chronic.

Small intestine diarrhea occurs in large volumes with a lack of urgency; the frequency of bowel movements does not always increase. Large intestine diarrhea, on the other hand, tends to occur in smaller volumes with urgency and increased frequency. Mucus and blood may be present. It is also possible for a dog to have mixed bowel diarrhea. All these observations highlight the difficulty in describing diarrhea, but are valuable for facilitating diagnostic and management decisions.

CAUSES OF DIARRHEA IN THE DOG Primary gastrointestinal causes of diarrhea include infectious disease (i.e. intestinal parasites, viruses), mechanical issues (i.e. foreign bodies, chronic intussusception), toxicities, noninfectious inflammatory problems and neoplasia.2 Non-infectious inflammatory problems include food sensitivities, dysbiosis, and idiopathic immune-mediated diseases.

Secondary gastrointestinal disorders include diseases affecting the liver, kidneys, exocrine pancreas, cardiovascular system, central nervous system, and endocrine glands.

The majority of CD cases in dogs result from dysregulated intestinal inflammation, currently termed chronic inflammatory enteropathy (CIE). For perspective, one report found that 90% of canine CD cases were primary enteropathies with the majority having an inflammatory basis (79%). Of those, 66% were food responsive, 23% were idiopathic, and 11% antibiotic responsive. Infectious disease was identified in only 13% of the primary enteropathy cases, followed by neoplasia at 4%. Only 10% of the total cases were diagnosed as secondary in origin.2 A similar pattern of distribution was found in other studies.3,4

FOCUS ON CHRONIC INFLAMMATORY ENTEROPATHY Chronic inflammatory enteropathy is common and illustrates important integrative approaches for patients with CD. Diagnosing CIE is a process of excluding other causes of diarrhea. A full workup should be performed, including a complete blood count, chemistry profile, T4, cortisol+/-ACTH stimulation, Creactive protein, trypsin-like immunoreactivity (TLI), pancreatic lipase immunoreactivity (PLI), folate, cobalamin, vitamin D, fecal flotation, and intestinal parasite antigen/PCR testing. Abdominal ultrasound should be considered to rule out other causes of CD.

Figure 1

It is important to note that a normal abdominal ultrasound does not rule out CIE. This means that for severely affected and poorly responsive patients, endoscopic or surgical biopsies may be necessary. Tests to evaluate the intestinal microbiota have become available and are a valuable part of patient assessment. Note that this process of patient evaluation should also allow for the diagnosis and management of CD that is not CIE (see Figure 1).

Case study – Echo Echo, a 2½-year-old neutered male Bernese Mountain Dog, had a history of chronic diarrhea, periodic vomiting, weight loss, otitis and pruritus. He was fully vaccinated and neutered as a puppy. He received multiple courses of antibiotics at two and four months of age for diarrhea and otitis, and prednisone at six months for otitis. His current caregiver adopted him with he was 20 months old. He tested positive for giardia on two occasions, and was treated with metronidazole and fenbendazole.

At 21 months of age, Echo was experiencing periodic bouts of diarrhea and vomiting, had mild otits, conjunctivitis, and pruritus. He was anxious and underweight with no abdominal discomfort. Spine misalignment was noted at the C1 and T12-L2 areas. His workup included the following with normal findings: complete blood count, chemistry profile, T4, TLI, PLI, cobalamin, folate, resting cortisol, fecal PCR for parasites and infective organisms, abdominal ultrasound, and thoracic radiographs. An upper GI endoscopy revealed a grossly normal esophagus and stomach. The duodenum was erythematous and friable. Histopathology indicated mild gastritis with lympoplasmacytic infiltration and enteritis with moderate lymphoplasmactic and eosinophilic infiltration. Serum allergy testing for environmental allergens indicated extensive sensitivity, as did saliva food sensitivity testing. The fecal microbiome was significantly imbalanced.

Integrative support included: osteopathic manipulations, laser therapy, acupuncture, Canine Dermal Support from Standard Process Veterinary Formulas, pre- and probiotics, glutamine, marshmallow root, pancreatic enzymes, Bach flower remedies, Pituitrophin PMG(R) from Standard Process, and low-dose budesonide.

Dietary changes were made multiple times in an effort to identify the ideal food for Echo, but given the saliva food sensitivity results, this has been difficult.

Outline of chronic diarrhea workup showing differentiation between primary and secondary enteropathies. This distinction helps define the patient support plan, as well as monitoring and prognosis. It is important to recognize that some patients with secondary enteropathies will benefit from the integrative support recommendations described for chronic inflammatory enteropathy.

Echo’s overall response to therapy has been good. At the 11-month follow-up, he was no longer experiencing diarrhea, vomiting, or otitis, and his pruritus was minimal.

He has gained weight to his ideal and is no longer anxious.

His gut microbiota is still not completely balanced, but efforts are ongoing.

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Assessing the initial severity of a dog’s CIE, and their response to therapy, is important. The canine inflammatory bowel disease activity index (CIBDAI) and the canine chronic enteropathy activity index (CCECAI) are useful tools for this assessment.5,6 They provide composite scores based on parameters like appetite, presence of vomiting, and stool consistency. CCECAI has additional parameters such as serum albumin levels and pruritus.

Figure 2

INTEGRATIVE MANAGEMENT OF CHRONIC DIARRHEA When the diagnostic workup has been completed, treating the identified cause(s) of primary and secondary enteropathies should begin (see Figure 2).

For example, patients with giardia should receive appropriate therapies. Patients with secondary enteropathies like liver disease should receive supportive care and medications and herbs with hepatic side effects should be discontinued, if appropriate. Dietary considerations need to be incorporated into the support plan; for example, a pancreatitis patient may benefit from a low-fat diet. Dogs with toxicities, foreign bodies, intestinal neoplasia, infectious diseases, or dietary indiscretions need to have these particular issues addressed. Keep in mind that these patients may still benefit from having their inflammatory responses modulated, as discussed below.

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An outline of the basic approach to integrative patient support care and management for inflammatory enteropathies. This approach uses multiple modalities to address contributors to intestinal inflammation. Each patient should be individually assessed and the integrative support plan based on that assessment. Not every patient needs all the supplements, herbs, and other treatments listed. Other herbs, including selected Chinese herbs, may be indicated. In some patients, the support plan may need to be implemented conservatively and gradually to avoid worsening the problem.

Recognize that each area listed here has a direct and indirect influence on some other facet of the inflammation contributors. The integrity of the epithelial barrier is impacted by the microbiota, nutrient deficiencies, ANS, and adrenal fatigue. These in turn affect immune regulation.

Figure 3

Most dogs with CD have a primary gastrointestinal issue with inflammation, and a dysregulated immune response. This means a coordinated, multiprong approach for managing the inflammation is critical. From an integrative perspective, support therapy should be directed at the contributors to intestinal inflammation, such as a disrupted epithelial barrier, dysbiosis, and abnormal mucosal immunity. Gut regulation and stress and anxiety management are also important (see Figure 3). Keep in mind that there is a dynamic interrelationship between these contributors that should be recognized.

A representation of the major components of, and contributors to, the complex pathogenesis of inflammatory intestinal disease. The interaction between genetic predisposition, dysregulated immune responses, and dysbiosis can lead to a chronically inflamed intestinal tract. Numerous other factors contribute to the development and propagation of an imbalanced inflammatory response and dysfunctional epithelial barrier. Effectively managing inflammatory disorders relies on addressing the contributors that propagate the inflammatory state.

DIETARY THERAPY, NUTRITIONAL SUPPLEMENTS, AND HERBS

Dietary therapy The majority of CIE patients have a food responsive enteropathy. This means all CIE dogs should be placed on an appropriate limited ingredient diet, a balanced home-prepared diet, or a hydrolyzed diet for at least two to four weeks, then reassessed.4 The selected diet can be continued another two to four weeks if improvement is noted. If it isn’t, the diet should be changed. During the food trial, regular monitoring and other supportive care should be provided as described below. Strict compliance with the specified food is critical. In addition, steps should be taken to manage nutritional deficiencies identified through the workup.

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Nutritional supplements •

eficiencies of cobalamin and folate have been associated D with chronic gastrointestinal disease. Hy-pocobalaminemia has been reported in 19% to 54% of dogs with CIE.7 These dogs may be predisposed to intestinal inflammation and may have a blunted response to other indicated therapies, such as hydrolyzed diets. Cobalamin is essential for mucosal regeneration and reduction of inflammatory infiltrates.

Remember, cobalamin deficiency may result from small intestine disease and/or dysbiosis.8

•

The majority of dogs with CIE have low serum vitamin D levels. Hypovita-minosis D is currently thought to contribute to the initiation of intestinal inflammation, based on the effect vitamin D has on modulating immune responses and inflammation. The severity of hypovitaminosis D correlates with clinical severity based on CIBDAI scoring and the outcome of therapy.9 Ideally, vitamin D supplementation should be based on serum level measurements, due especially to the variability in patient baseline levels and variations in patient ability to absorb orally administered vitamin D. Follow-up serum measurements should be performed at eight- to ten-week intervals to determine if appropriate serum levels have been achieved and hypervitaminosis D avoided.10

•

Support for the intestinal epithelial barrier should be incorporated into the therapeutic plan. This includes the use of glutamine, which plays an important role in the health and metabolism of intestinal epithelial cells. Glutamine promotes intestinal epithelial cell proliferation, has antiinflammatory effects, and regulates tight junction proteins.

The intestine uses about 30% of the body’s total glutamine, and as much as 75% of ingested glutamine.11 In addition, the large intestine epithelial barrier can be supported through microbiota fermentation of soluble fiber and supplements

like arabinogalactan. These are used to increase the levels of short chain fatty acids (SCFA), which can improve the integrity of the large intestine epithelial barrier and offer immune modulating effects that promote reduced inflammation.4,12 •

Reactive oxygen species (ROS) and reactive nitrogen species (RNS) can be a major factor in initiating and contributing to the progression of inflammation in the gastrointestinal tract.13 They have direct cell membrane-damaging effects that increase cell permeability, cause DNA damage, and induce apoptosis. In addition, ROS and RNS upregulate genes associated with innate and adaptive immunity in the gastrointestinal tract.14 The intestinal tract is a significant source of reactive species. While some level of reactive species contributes to health, their continuous excess production and inadequate removal can play a notable role in promoting chronic inflammation. The addition of whole food ingredients with antioxidant properties, such as polyphenolic compounds and substances like carotenoids, can be beneficial.14 Polyphenols include flavonoids such as anthocyanins, and nonflavonoids such as stilbenes.

Cautious use of food ingredients is warranted to avoid aggravating potential food sensitivities associated with CIE. Improvements in the oxidative status of the gut can lead to beneficial changes in the microbiota.

Herbs Herbs like curcumin (Curcuma longa), ginger (Zingiber officinale) and green tea (Camellia sinensis) extracts could be considered for their impacts as antioxidants.14,15 However, many herbs also have anti-inflammatory properties that can help manage inflammation associated with CIE. Herbs such as marshmallow root (Althaea officinalis) have antioxidant properties, antiinflammatory effects, and the added advantage of providing

Managing the central and autonomic nervous systems Stress and anxiety contribute to disruptions in the regulation of gastrointestinal motility and secretions, reduced integrity of the epithelial barrier, and the release of inflammatory mediators.

This influence occurs through the autonomic nervous system (ANS) and intrinsic enteric nervous system. Actions like the release of corticotrophin-releasing factor can result in increased mucosa permeability and mast cell degranulation. Stress-related inhibition of parasympathetic nervous system input reduces its anti-inflammatory effect.20 Increased permeability allows more interface with a disrupted microbiota, further complicating management of intestinal inflammation. Managing adrenal fatigue may be an important part of addressing the stress response.

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Even though the intrinsic enteric nervous system is focused on proper regulation of the digestive process, it can be influenced by the ANS. In addition to affecting gastrointestinal motility, mucosal secretions, and vascular tone, it also influences intestinal epithelial proliferation and intestinal inflammation.

ANS dysfunction has been associated with inflammatory intestinal disease.21 The main parasympathetic innervation occurs through the vagus nerve, and the sympathetic via the splanchnic nerves. Appropriate osteopathic manipulation of the associated spinal column segments, along with acupuncture, appear to be valuable for balancing the respective signaling and restoring appropriate function.

a protective mucilaginous coating for inflamed mucosa.16 It is important to be aware of the potential side effects of herbs, and manage them accordingly. For example, ginger can have impacts on platelets and clotting, while marshmallow root can interfere with the absorption of medications. Carefully selected Chinese herbs can also be beneficial.

INTESTINAL DYSBIOSIS MANAGEMENT The intestinal microbiota is a complex ecosystem that influences health in numerous ways. Studies show a link between CIE, intestinal dysbiosis, and intestinal inflammation.17 This is likely related to the interaction between the digestive process, ingesta, and microbiota in genetically susceptible individuals.18 A full discussion of the role of microbiota in inflammatory enteropathies, and their management, is beyond the scope of this article. However, brief mention will be made with the intention of focusing on supportive therapy for patients with CD and CIE. Managing intestinal dysbiosis can be challenging and often requires a multistep approach. Optimally, an evaluation of the microbiota using current molecular sequencing techniques should be done. This will facilitate an understanding of how significant the dysbiosis imbalance is for that patient; identify specific approaches for addressing the imbalance; and provide a way to monitor the impact of therapy on the microbiota.

Adjustments to ongoing supportive therapy can be made based on serial evaluation of the microbiota.

Managing dysbiosis includes dietary changes, food additives, the use of probiotics, fecal transfer, administration of prebiotic substances, addition of organisms known to suppress pathogens, and the identification of factors that impact the digestive tract microenvironment. When making changes in the dog’s diet, or incorporating additives to the diet, be aware of potential impacts

on food responsive disorders or other clinical conditions and avoid changes that can be problematic. Dietary changes that may be beneficial for altering the microbiota include adding protein toppers and reducing carbohydrates. Increasing fiber can also contribute to needed microbiota changes. Adding food ingredients and herbs with antioxidant properties that contribute to the reduction of ROS and RNS can promote important alterations.

An overwhelming range of choices and recommendations for probiotics is available. Considerations include numbers of colony forming units (CFU), numbers of microorganism species, types of organisms (i.e. soil microbes, animal origin, etc), and inclusion of prebiotic compounds. Some authorities recommend one to ten billion CFU per day, but this can depend on the individual patient. Select products that give consistent patient results based on clinical response and monitoring of the microbiota. Sometimes, a combination of products is necessary and includes prebiotic substances like fructooligosaccharide (FOS). These substances can aid in selective proliferation of beneficial microbiota organisms, increase production of SCFA, improve the epithelial barrier, and promote anti-inflammatory processes.19 The microenvironment of the gastrointestinal tract can contribute to dysbiosis and have a negative impact on achieving desired changes. While altering the diet, incorporating additives, and using probiotics contribute to changing the microenvironment, it is important to recognize that the microenvironment is also impacted by conditions related to the pH of gastric fluid; the quantity and quality of pancreatic fluid, including digestive enzymes; bile acid secretions; and the regular motility of the digestive tract. Herbs may promote beneficial changes in the microenvironment. For example, milk thistle (Silybum marianum) and dandelion (Taraxacum officinale) can promote increased bile production.

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The enteric and autonomic nervous systems (see sidebar on page 22) have impacts on the microenvironment of the digestive tract through their influence on motility, digestive secretions, the epithelial barrier, and immune regulation. It’s important to address management of the negative factors that reduce or limit their regulatory function. This includes reducing stress and anxiety, treating spinal column misalignment, and using modalities like acupuncture.

Note that the use of antibiotics for patients with CD is being debated. Antibiotics may contribute to long term problems with dysbiosis. Patients that respond to antibiotic therapy may already have significant dysbiosis. However, some patients may need a long course of antibiotics while other integrative support options are being implemented.

INTESTINAL MUCOSA SUPPORT The intestinal mucosa should be supported as outlined above, including supplements like glutamine, arabinogalactan, antioxidants, and anti-inflammatory herbs. Managing stress, anxiety, and autonomic imbalances plays an important role in the health of the mucosa and immune regulation.

SUMMARY Chronic diarrhea in dogs can be a complex and challenging problem to resolve. A complete workup and understanding of the precipitating and contributing factors is essential, along with recognizing that the majority of CD patients have a dysregulated immune response that propagates intestinal inflammation. Optimal support for these patients relies on a methodical and encompassing integrative treatment plan.

This is especially true because of the interrelated nature of the contributing factors. Serial monitoring and therapy adjustments are also important for optimal success.

Ivasovic F, Ruetten M, Kook PH. Prevalence of inflammatory versus neoplastic lesions in dogs with chronic gastrointestinal signs undergoing gastroduodenoscopy: 195 cases (2007–2015). Res Vet Sci. 2022;146:28-33.

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Jergens AE, Heilmann RM. Canine chronic enteropathy – current state-of-the-art emerging concepts. Front Vet Sci. 2022 Sep 21;9:923013. doi: 10.3389/fvets.2022.923013. PMID: 36213409; PMCID: PMC9534534.

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Jergens AE, Schreiner CA, Frank DE, et al. A scoring index for disease activity in canine inflammatory bowel disease. J Vet Intern Med. 2003;17:291–297.

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Allenspach K, Wieland B, Gröne A, et al. Chronic enteropathies in dogs: evaluation of risk factors for negative outcome. J Vet Intern Med. 2007;21:700–708.

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Heilmann RM, Steiner JM. Clinical utility of currently available biomarkers in inflammatory enteropathies of dogs. J Vet Intern Med. 2018;32:1495–1508.

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Kather S, Grützner N, Kook PH, et al. Review of cobalamin status and disorders of cobalamin metabolism in dogs. J Vet Intern Med. 2020;34:13–28.

8

Titmarsh H, Gow AG, Kilpatrick S, et al. Association of vitamin D status and clinical outcome in dogs with a chronic enteropathy. J Vet Intern Med. 2015;29:1473–1478.

9

VDI Laboratory, LLC. Vitamin D growing body of evidence 2021. https://vdilab.com/wp-content/ uploads/2021/09/2021-Body-of-Evidence_Vitamin-D.pdf accessed 5-4-23.

10

Kim MH, Kim H. The roles of glutamine in the intestine and its implication in intestinal diseases. Int J Mol Sci. 2017;18:1051; doi:10.3390/ijms18051051.

11

Pérez-Reytor D, Puebla C, Karahanian E, et al. Use of short-chain fatty acids for the recovery of the intestinal epithelial barrier affected by bacterial toxins. Front Physiol. 2021; May 24;12:650313. doi: 10.3389/ fphys.2021.650313. PMID: 34108884; PMCID: PMC8181404.

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Dziąbowska-Grabias K, Sztanke M, Zając P, Celejewski M, Kurek K, Szkutnicki S, Korga P, Bulikowski W, Sztanke K. Antioxidant therapy in inflammatory bowel diseases. Antioxidants (Basel). 2021 Mar 9;10(3):412. doi: 10.3390/antiox10030412. PMID: 33803138; PMCID: PMC8000291.

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Tian T, Wang Z, Zhang J. Pathomechanisms of oxidative stress in inflammatory bowel disease and potential antioxidant therapies. Oxid Med Cell Longev. 2017;2017:4535194. doi: 10.1155/2017/4535194. Epub 2017 Jun 28. PMID: 28744337; PMCID: PMC5506473.

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Sadeghi Poor Ranjbar F, Mohammadyari F, Omidvar A, et al. Zingiber officinale (Ginger) as a treatment for inflammatory bowel disease: A review of current literature. Front Drug Discov. 2022; 2:1043617. doi: 10.3389/ fddsv.2022.1043617.

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Bonaterra GA, Bronischewski K, Hunold P, et al. Anti-inflammatory and anti-oxidative effects of Phytohustil R and root extract of Althaea officinalis L. on macrophages in vitro. Front Pharmacol. 2020; 11:290. doi: 10.3389/ fphar.2020.00290.

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Honneffer JB, Minamoto Y, Suchodolski JS. Microbiota alterations in acute and chronic gastrointestinal inflammation of cats and dogs. World J Gastroenterol. 2014;Nov 28;20(44):16489-97. doi: 10.3748/wjg.v20. i44.16489. PMID: 25469017; PMCID: PMC4248192.

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Disclosure: The author of this article formulated and clinically evaluated the original 16 Standard Process Veterinary Formulas™ during product development as a paid consultant for Standard Process Inc.® He is not employed by Standard Process Inc.®, derives no financial benefit and has no equity interest in either Standard Process Inc.® or Standard Process Veterinary Formulas™.

Marchetti V, Gori E, Mariotti V, et al. The impact of chronic inflammatory enteropathy on dogs’ quality of life and dog-owner relationship. Vet Sci. 2021 Aug 19;8(8):166. doi: 10.3390/vetsci8080166. PMID: 34437488; PMCID: PMC8402703.

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Volkmann M, Steiner JM, Fosgate GT, et al. Chronic diarrhea in dogs -- retrospective study in 136 cases. J Vet Intern Med. 2017;31:1043-1055.

2

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Simpson KW, Jergens AE. Pitfalls and progress in the diagnosis and management of canine inflammatory bowel disease. Vet Clin North Am Small Anim Pract. 2011 Mar;41(2):381-98. doi: 10.1016/j.cvsm.2011.02.003.

PMID: 21486642.

18

Roy S, Dhaneshwar S. Role of prebiotics, probiotics, and synbiotics in management of inflammatory bowel disease: Current perspectives. World J Gastroenterol. 2023;Apr 14;29(14):2078-2100. doi: 10.3748/wjg.v29. i14.2078. PMID: 37122604; PMCID: PMC10130969.

19

Sun Y, Li L, Xie R, et al. Stress Triggers Flare of Inflammatory Bowel Disease in Children and Adults. Front Pediatr.

2019;7:432. doi:10.3389/fped.2019.00432.

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Duan H, Cai X, Luan Y, et al. Regulation of the autonomic nervous system on intestine. Front Physiol.

2021;12:700129. doi: 10.3389/fphys.2021.700129.

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ADVERTORIAL

Support the Gut and the Body Will Follow:

Digestive Health is Key to

Whole Body Wellness GUT STATUS AT BIRTH The gut microbiome is not technically an organ, but it does have important functions in puppies and kittens, including training the immune system.1 A balanced microbiome primes and stimulates the immune system, aids in defense against intestinal pathogens, and provides diverse health benefits to the body.2-4 Unfortunately, newborn mammals like puppies and kittens are born with an underdeveloped gut microbiome and low immunity, which increases their susceptibility to invading pathogens.1,4 The nutritional status of the mother and the immunomodulating nutrients they receive from colostrum largely determine the bacteria that colonize before and during birth.1,5,6

DOWNFALLS OF POOR GUT HEALTH DURING GROWTH The growing period is critical to longterm health in dogs and cats.1 The gut microbiome is more sensitive to disruptions during this time, including those caused by antibiotics. Negative changes in the microbiota can cause issues later in life, such as increased metabolic and immune disorders in animals.1,6,7 Because puppies in particular often experience acute diarrhea, it is especially important to support the development of a healthy gut microbiome instead of relying on antibiotics.6 Similarly, studies have shown that treatment with antibiotics in kittens can delay maturation of the gut microbiome for three months.8

BY KERI BARRON, PHD

PROACTIVE SUPPORT OF GUT HEALTH IN PUPPIES AND KITTENS The early developmental window presents a unique opportunity to intervene.

Interventions seeking to modulate the gut microbiota in puppies may be more effective in establishing a healthy gut than those undertaken in adult dogs.9 Because the brain and immune system are still developing in the postnatal period, the nutritional environment can either support or disrupt the maturation and development processes.1,6 Experiences and exposures during critical periods of development can have cumulative effects on behavioral traits, cognitive abilities, trainability, health, and performance.6 As such, nutritional interventions that optimize the composition and function of the microbiome can improve the health of both cats and dogs and support their over-all digestive and immune health.

Whereas antibiotics and environmental stress can cause dysbiosis in newborn puppies and kittens, nutritional support can positively affect gut microbiome health by helping to balance bacterial species.1 Supplementation with a natural plant polyphenol in puppies alleviates oxidative stress and inflammation associated with environmental stress by supporting the gut microbiome.10 Additionally, prebiotics, probiotics, and postbiotics can help support a healthy gut microbiome.2,11 Finally, providing a diet that meets nutritional requirements, as well as enhances the function of the

microbiome, can significantly improve and support the holistic health of cats and dogs.

Keri Barron, PhD, is the Scientific Nutrition Writer for Standard Process Inc. located at the Nutrition Innovation Center in Kannapolis, North Carolina. Her work involves creating educational materials and translating scientific articles for audiences to support health and wellness.

Keri has a B.S. in Nutritional Biochemistry and Metabolism and M.S. in Nutrition from Case Western Reserve University in Cleveland, Ohio. She continued her studies at the Nutrition Research Institute, a remote campus of the University of North Carolina at Chapel Hill, earning a PhD in Nutrition.

Garrigues, Q., et al. (2022). Gut microbiota development in the growing dog: A dynamic process influenced by maternal, environmental and host factors. Front Vet Sci, 9:964649.

1

Ziese, A.L., Suchodolski, J.S. (2021). Impact of Changes in Gastrointestinal Microbiota in Canine and Feline Digestive Diseases. Vet Clin N Am-Small, 51:155-169.

2

P etersen, C., Round, J.L. (2014). Defining dysbiosis and its influence on host immunity and disease. Cell Microbiol, 16(7):1024-1033.

3

S uchodolski, J.S. (2014). Companion animals symposium: microbes and gastrointestinal health of dogs and cats. J Anim Sci, 89:1520-1530.

4

Chastant, S., Mila, H. (2019). Passive immune transfer in puppies. Anim Reprod Sci, 207:162.

5

G aillard, V., et al. (2022). Environmental risk factors in puppies and kittens for developing chronic disorders in adulthood: A call for research on developmental programming. Front Vet Sci, 9:944821.

6

B oulangé, C.L., et al. (2016). Impact of the gut microbiota on inflammation, obesity, and metabolic disease. Genome Med, 8(1):4253.

7

Stavroulaki, E., et al. (2021). Short- and long-term effects of amoxicillin/ clavulanic acid or doxycycline on the gastrointestinal microbiome of growing cats. Plos ONE, 16:e0253031

8

Pereira, A.M., Clemente, A. (2021). Dogs’ Microbiome From Tip to Toe. Top Companion Anim Med, 45:100584.

9

Yang, K., et al. (2022). Gallic Acid Alleviates Gut Dysfunction and Boosts Immune and Antioxidant Activities in Puppies Under Environmental Stress Based on Microbiome-Metabolomics Analysis. Front Immunol, 12:813890.

10

ernimont, S.M., et al. (2020). The Effects of Nutrition on the W Gastrointestinal Microbiome of Cats and Dogs: Impact on Health and Disease. Front Microbiol, 11:1266-1289.

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A CLOSER LOOK AT THE LINK BETWEEN EPIGENETICS AND CANINE CANCER

BY KELLY DIEHL, DVM, MS

Understanding the epigenetic changes that occur in canine cancer can provide insights into the underlying mechanisms of the disease, and help identify new targets for treatment.

Cancer is a complex disease, and despite decades of research, it remains a leading cause of death in dogs over two years old. We know cancer is not a monolithic disease, and that its development is influenced by many external factors, such as genetics and the environment. Mounting evidence suggests that epigenetics also plays a role in cancer development and progression. In this article, we’ll take a closer look at what we know about the relationship between canine cancer and epigenetics.

WHAT IS EPIGENETICS? Epigenetics studies the changes in gene expression that occur without altering the DNA sequence. This is important to remember, and is different from, for example, a mutation that alters DNA sequence (such as a deletion or frame shift).

A variety of environmental factors, such as diet, exercise, stress, and exposure to toxins, can alter gene expression.

Epigenetic changes also can be inherited from one generation to the next.

Epigenetic changes can affect gene expression by altering the structure of DNA, modifying histones (proteins that help package DNA into chromatin), and altering the expression of non-coding RNAs (more on this later).

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Before we go much further, we probably need a quick refresher on DNA structure and basic molecular biology, in order to understand epigenetics.

1

F irst, we need to remember something called the central dogma of molecular biology: DNA contains the information to code all the proteins in the body, and RNA is the messenger that transmits this information to the ribosomes responsible for making proteins.

2

e must also precisely define what we mean by gene W expression. This term refers to the process involved in turning the DNA code contained in a particular unit identified as a gene into a “product” — typically a protein but also RNA.

3

e should recall from basic biology class that two W processes — transcription and translation — are involved in gene expression. Transcription refers to the conversion of DNA code to RNA, and translation is the process occurring in the ribosomes where the information coded by the RNA is used to direct the formation of new proteins.

4

astly, it’s important to remember that gene expression L is regulated — it doesn’t just “happen.” Factors that can affect gene expression include cellular signals, feedback mechanisms, and the environment.

WHAT DOES EPIGENETICS HAVE TO DO WITH CANCER? In cancer cells, epigenetic changes can alter the expression of genes that regulate cell growth and division, leading to uncontrolled cell growth and the formation of tumors. Epigenetic changes can also affect the ability of cancer cells to invade surrounding tissues and spread to other parts of the body.

One of the most well-studied epigenetic changes in cancer is DNA methylation. As the name implies, DNA methylation refers to the addition of a methyl group to a cytosine nucleotide in a DNA molecule, but also can refer to a loss of a methyl group in other regions. Methylation affects gene expression by altering the accessibility of DNA to transcription factors and other regulatory proteins. In cancer cells, the addition of a methyl group in one location, and loss in another, can result in the silencing of tumor suppressor genes and the activation of oncogenes, promoting tumor growth and progression. As a side note, methylation is also associated with aging and a lot of research is looking at DNA methylation as a marker of unhealthy aging and disease risk.

Histone modifications are another important epigenetic change in cancer. Histones are proteins that help package DNA into chromatin. Modifications to histones, including acetylation and methylation, can once again affect the accessibility of DNA to transcription factors and other regulatory proteins, with the same outcomes noted previously.

Earlier, we talked about the central dogma of DNA to RNA to proteins. However, not all RNA ends up translated into protein; this type of RNA is called non-coding RNA. The term “noncoding” is a bit of a misnomer, since non-coding RNAs, such

as microRNAs and long non-coding RNAs, are also important regulators of gene expression. These RNAs can interact with messenger RNAs (mRNAs) to inhibit their translation into proteins or target them for degradation. In cancer cells, noncoding RNAs can be dysregulated, leading to the altered expression of genes that regulate cell growth and division.

WHAT TYPES OF ENVIRONMENTAL FACTORS CAUSE EPIGENETIC CHANGES? A growing body of research in human medicine looks at how the environment influences epigenetic changes.

The most obvious factor is simply aging. During fetal development, epigenetic changes dictate which cells become nerve cells, muscle cells or heart cells. As we (and our dogs) age, we’re exposed to lots of substances that have the potential to cause changes. In addition, certain epigenetic changes seem to be a part of normal aging.

Several lifestyle factors have been implicated in epigenetic changes and health outcomes (positive and negative), although we need to remember that some are just associated with disease and others have only been demonstrated in vitro.

These factors include obesity (negative), exercise (positive), air pollution (negative), smoking (negative), polyphenols in foods (positive), alcohol consumption (negative) and emotional stress (negative).1 An intriguing — and important — fact about epigenetic changes is that they can influence the health of future generations.

An often-cited example from human medicine comes from studies of Dutch women who were pregnant during the winter famine of 1944-1945. Over 60 years later, researchers found that these children, now adults, had a higher incidence of certain diseases. Further examination revealed these individuals had higher levels of methylation at some gene loci when compared to siblings not affected by the famine.2-5 There is also evidence that epigenetic changes are reversible.

Another example from human medicine comes from studies of smokers. There is plenty of evidence that smoking triggers a variety of epigenetic changes, but when people stop smoking, many of these epigenetic changes reverse themselves.6 Unfortunately, things are a bit murkier when it comes to environmental factors, epigenetic changes, and canine cancer.

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LIFESTYLE FACTORS ASSOCIATED WITH EPIGENETIC CHANGES IN HUMANS • Diet/nutrition • E xercise • Obesity

• Smoking

• P ollutants • A lcohol

consumption

• S tress

PRIMARY MECHANISMS OF EPIGENETIC CHANGES • Altering • Histone DNA alterations through methylation

• Non-coding RNA

CANCERS ASSOCIATED WITH EPIGENETIC CHANGES • Mammary cancer • Melanoma

• Osteosarcoma

• H emangiosarcoma • H igh grade mast cell tumors

ADDITIONAL RESOURCES Horvath S, Lu AT, Haghani A, et al. DNA methylation clocks for dogs and humans. Proc Natl Acad Sci USA(2022) 119(21) doi:10.1073/pnas.2120887119:An excellent review of DNA methylation and aging.

Mantaner-Angoiti E, Marin-Garcia PJ, Llobat L. Epigenetic alterations in canine malignant lymphoma: future and clinical outcomes. Animals(2023) 13:468488. doi:10.3390/ani13030468: A great review of epigenetics and canine lymphoma.

Xavier PLP, Muller S, Fukumasu H. Epigenetic mechanisms in canine cancer. Front Oncol (2020) 10:591843 doi: 10.3389/onc.2020.591843 — Another excellent, in-depth discussion of canine cancer and epigenetics.

The Centers for Disease Control and Prevention website (cdc.gov) has a nice section on epigenetics and human health. Some of their examples are cited in this article.

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insecticides, pesticides and herbicides have all been linked to cancer development in dogs, but the exact mechanisms remain unknown.

EPIGENETICS AND CANINE CANCER Researchers have a growing interest in studying epigenetic changes and their influence on the development of cancer in dogs. Understanding these changes could point to new treatments and possibly even preventive measures for many different types of canine cancer.

DNA methylation patterns are an active area of research.

Although a definitive link between methylation patterns and alterations, and cancer development, hasn’t been established, an accumulating body of evidence shows that this process is important.

Several published studies have linked certain hypomethylation patterns to the development of lymphoma, leukemia, high-grade mast cell tumors, and lung cancer. Conversely, hypermethylation of other regions also have been associated with lymphoma, leukemia, and melanoma in dogs.7-17 Histone modifications also have been linked to canine cancer, specifically bladder cancer and osteosarcoma.18-22 Currently, Morris Animal Foundation has one active grant studying a mutation in the SETD2 gene, which encodes a histone methyltransferase. In this instance, methylation is important for normal function, and if mutated, the dysregulation is associated with aggressive osteosarcoma in people. This Foundation-funded study is looking at the same mutation in dogs, and determining if it could ultimately be leveraged as a therapeutic target.

Non-coding RNAs also are implicated in cancer development. Most veterinarians are probably familiar with microRNAs and their association with cancer. Evidence exists for microRNA dysregulation in osteosarcoma, mast cell tumors, lymphoma, mammary cancer, melanoma, and hemangiosarcoma. MicroRNAs in the blood and urine have generated a lot of interest as cancer biomarkers that could be used as screening tools.23-30

NEW EPIGENETIC RESEARCH FUNDED BY MORRIS ANIMAL FOUNDATION In the last five years, the Foundation has seen a big uptick in the number of proposal submissions focused on epigenetics.

Current projects in progress or under consideration include:

9 Epiphanio TMF, de Azevedo Fernandes NCC, de Oliveira TF, Lopes PA, Réssio RA, Gonçalves S, et al.

Global DNA methylation of peripheral blood leukocytes from dogs bearing multicentric non-Hodgkin lymphomas and healthy dogs: A comparative study. PloS One (2019) 14:1–22. doi: 10.1371/journal. pone.0211898.

10 Herrera CL, Kim DY, Kumar SR, Bryan JN. Peroxisome proliferator activated receptor γ protein

expression is asymmetrically distributed in primary lung tumor and metastatic to lung osteosarcoma samples and does not correlate with gene methylation. BMC Vet Res (2015) 11:1–11. doi: 10.1186/ s12917-015-0547-x.

11Shi H, Guo J, Duff DJ, Rahmatpanah F, Chitima-Matsiga R, Al-Kuhlani M, et al. Discovery of novel

epigenetic markers in non-Hodgkin’s lymphoma. Carcinogenesis (2007) 28:60–70. doi: 10.1093/ carcin/bgl092

12Bryan JN, Taylor KH, Henry CJ, Selting KA, Rahmatpanah F, Lewis MR, et al. DNA methylation in

• S tudying methylation for early detection and recurrence of canine hemangiosarcoma

cancer: techniques and preliminary evidence of hypermethylation in canine lymphoma. Cancer Ther (2008) 6:137–48.

• Using a DNA methylation clock to study aging in dogs

deoxycitidine in melanoma cells: Demethylation of TNF-α and activation of FOXO1. Cancer Lett (2015) 369:344–53. doi: 10.1016/j.canlet.2015.08.023.

13Noguchi S, Mori T, Igase M, Mizuno T. A novel apoptosis-inducing mechanism of 5-aza-2’-

• S tudying the therapeutic utility of SETD2 mutations in canine osteosarcoma

14Noguchi S, Mori T, Nakagawa T, Itamoto K, Haraguchi T, Mizuno T. DNA methylation contributes

• Studying changes associated with aging of the canine immune system.

15Sato M, Mochizuki H, Goto-Koshino Y, Fujiwara-Igarashi A, Takahashi M, Ohno K, et al. Prognostic

toward silencing of antioncogenic microRNA-203 in human and canine melanoma cells. Melanoma Res (2015) 25:390–8. doi: 10.1097/CMR.0000000000000183 significance of hypermethylation of death-associated protein kinase (DAPK) gene CpG island in dogs with high-grade B-cell lymphoma. Vet Comp Oncol (2018) 16:409–15. doi: 10.1111/vco.12395.

16Bronzini I, Aresu L, Paganin M, Marchioretto L, Comazzi S, Cian F, et al. DNA methylation and

We’ve even funded a few cat and wildlife epigenetic studies that could have implications for dogs as well.

The Golden Retriever Lifetime Study was initiated as a cancer risk factor project. A current project involves testing urine and blood samples from dogs diagnosed with lymphoma for evidence of environmental toxin exposure.

Understanding the epigenetic changes that occur in canine cancer can provide valuable insights into the underlying mechanisms of the disease, and help identify new targets for cancer treatment. Epigenetic therapies, such as DNA methyltransferase inhibitors and histone deacetylase inhibitors, have shown promise in human cancer treatment and may also be effective in the treatment of canine cancer.

targeted sequencing of methyltransferases family genes in canine acute myeloid leukaemia, modelling human myeloid leukaemia. Vet Comp Oncol (2017) 15:910–8. doi: 10.1111/vco.12231.

17Yamazaki J, Jelinek J, Hisamoto S, Tsukamoto A, Inaba M. Dynamic changes in DNA methylation

patterns in canine lymphoma cell lines demonstrated by genome-wide quantitative DNA methylation analysis. Vet J (2018) 231:48–54. doi: 10.1016/j.tvjl.2017.11.007.

18 Eto S, Saeki K, Yoshitake R, Yoshimoto S, Shinada M, Ikeda N, et al. Anti-tumor effects of the histone

deacetylase inhibitor vorinostat on canine urothelial carcinoma cells. PloS One (2019) 14:1–15. doi: 10.1371/journal.pone.0218382.

19Sakthikumar S, Elvers I, Kim J, Arendt ML, Thomas R, Turner-Maier J, et al. SETD2 is recurrently

mutated in whole-exome sequenced canine osteosarcoma. Cancer Res (2018) 78:3421–31. doi: 10.

1158/0008-5472.CAN-17-3558.

20 Gardner HL, Sivaprakasam K, Briones N, Zismann V, Perdigones N, Drenner K, et al. Canine

osteosarcoma genome sequencing identifies recurrent mutations in DMD and the histone methyltransferase gene SETD2. Commun Biol (2019) 2:1–13. doi: 10.1038/s42003-019-0487.

21Choi HJ, Jang S, Ryu JE, Lee HJ, Lee HB, Ahn WS, et al. Significance of EZH2 expression in canine

mammary tumors. BMC Vet Res (2016) 12:1–7. doi: 10.1186/S12917-016-0789-2.

22Choi HJ, Lee HB, Park HK, Cho SM, Han HJ, Lee SJ, et al. EZH2 expression in naturally occurring canine

tumors. Comp Med (2018) 68:148–55.

23Boggs RM, Wright ZM, Stickney MJ, Porter WW, Murphy KE. MicroRNA expression in canine

mammary cancer. Mamm Genome (2008) 19:561–9. doi: 10.1007/s00335-008-9128-7.

24Fenger JM, Bear MD, Volinia S, Lin TY, Harrington BK, London CA, et al. Overexpression of miR-9 1Alegria-Torres JA, Baccarelli A, Bollati V. Epigenetics and lifestyle. Epigenomics (2011) 3(3):267-277.

doi:10.2217/EPI.11.22.

2Roseboom T. Epidemiological evidence for the developmental origins of health and disease: effects

of prenatal undernutrition in humans. J Endocrinol (2019) 242:T135-T144.

3Heijmans B, Tobi E, Stein A, et al. Persistent epigenetic differences associated with prenatal exposure

to famine in humans. Proc Natl Acad Sci U S A (2008) 105:17046-17049

in mast cells is associated with invasive behavior and spontaneous metastasis. BMC Cancer (2014) 14:1–16. doi: 10.1186/1471-2407-14-84.

25Fenger JM, Roberts RD, Iwenofu OH, Bear MD, Zhang X, Couto JI, et al. MiR-9 is overexpressed in

spontaneous canine osteosarcoma and promotes a metastatic phenotype including invasion and migration in osteoblasts and osteosarcoma cell lines. BMC Cancer (2016) 16:1–19. doi: 10.1186/ s12885-016-2837-5.

26Grimes JA, Prasad N, Levy S, Cattley R, Lindley S, Boothe HW, et al. Smith BF. A comparison of

Common and Timing- And Sex- Specific. Hum Mol Genet (2009) 18:4046-53.

microRNA expression profiles from splenic hemangiosarcoma, splenic nodular hyperplasia, and normal spleens of dogs. BMC Vet Res (2016) 12:1–12. doi: 10.1186/s12917-016-0903-5.

5Tobi E, Slieker R, Luijk R, et al., DNA methylation as a mediator of the association between prenatal

27Kobayashi M, Saito A, Tanaka Y, Michishita M, Kobayashi M, Irimajiri M, et al. Microrna expression

6McCartney D, Stevenson A, Hillary R, et al. Epigenetic signatures of starting and stopping smoking.

28 Craig KKL, Wood GA, Keller SM, Mutsaers AJ, Wood RD. MicroRNA profiling in canine multicentric

7Pelham JT, Irwin PJ, Kay PH. Genomic hypomethylation in neoplastic cells from dogs with malignant

29 Rahman MM, Lai YC, Husna AA, Chen HW, Tanaka Y, Kawaguchi H, et al. Micro RNA transcriptome

8 Morimoto CY, Tedardi MV, da Fonseca IIM, Kimura KC, Sanches DS, Epiphanio TF, et al. Evaluation

30 Ushio N, Rahman MM, Maemura T, Lai YC, Iwanaga T, Kawaguchi H, et al. Identification of

4Tobi E, Lumey L, Talens R, et al., DNA Methylation Differences After Exposure to Prenatal Famine Are

adversity and risk factors for metabolic disease in adulthood. Sci Adv (2018) 4:eaao4364.

EBioMedicine (2018) 37:214-220.

lymphoproliferative disorders. Res Vet Sci (2003) 74:101–4. doi: 10.1016/S0034-5288(02)00179-0.

of the global DNA methylation in canine mast cell tumour samples by immunostaining of 5-methyl cytosine. Vet Comp Oncol (2017) 15:1014–8. doi: 10.1111/vco.12241.

profiling in canine prostate cancer. J Vet Med Sci (2017) 79:719–25. doi: 10.1292/jvms.16-0279. lymphoma. PloS One (2019) 14:1–24. doi: 10.1371/journal.pone.0226357.

profile in canine oral melanoma. Int J Mol Sci (2019) 20:1–19. doi: 10.3390/ijms20194832.

dysregulated microRNAs in canine malignant melanoma. Oncol Lett (2019) 17:1080–8. doi: 10.3892/ ol.2018.9692.

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industry innovations Sustainable quality nutrition Premium nutrition, sustainable sourcing, and high palatability are now available in a scoop-and-serve format. Earth Animal’s Wisdom Air-Dried dog food uses proprietary airdrying technology to ensure the highest nutrient retention and flavor in an easy-to-serve format. Wisdom includes combinations of sustainably-sourced meat, organs, seafood, ancient grains, sprouted seeds, fruits, and vegetables. Perfect as a mixer, topper or treat, and packed in one of the industry’s first recyclable bags. https://earthanimal.com/our-products/wisdom-dog-food/

Boost their brain health Companion dogs have long been recognized for their ability to improve mental health in humans.

But what about their own mental well-being? ThorneVet Canine Cognitive Support helps with neurological health in dogs. It combines nutrients, antioxidants, phospholipids, and botanicals in a multi-modal formula designed to enhance cerebro-cortical blood flow, optimize circulation and oxygen utilization, and nourish the nervous system. This powerhouse of therapeutic ingredients offers robust support for the aging dog’s nervous system, and protection against degenerative disorders. https://thornevet.com/product/canine-cognitive-support/

New technique accurately identifies salmon in food products Salmon is a high-value commodity with a frequently complicated supply chain in the fisheries sector, making it particularly susceptible to mislabelling and other forms of fraud. Developing dependable techniques for determining the authenticity of salmon in commercial foods has been challenging, but a study done by Queen's University Belfast's Institute for Global Food Security (IGFS) has led to the development of a method that uses AI to identify genuine salmon with 100% accuracy.

Over 500 samples of wild and farmed salmon were collected from four of the world's major salmon-producing regions: Scotland, Alaska, Norway, and Iceland.

The fishes’ elemental fingerprint and lipid fingerprint measurements were combined to create a single “data lake.” Powerful data analytics were then utilized to create a user-friendly diagnostic tool that identifies a fish's origins (wild or farmed) and mode of production.

In order to compare results, more salmon samples from UK shops were analyzed at the conclusion of the trial. Again, there was perfect correlation between the data provided by the merchants and what the researchers discovered in the lab. www.nature.com/articles/s41467-023-38382-z

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Tracker helps keep pets safe and healthy Your clients’ main priority is keeping their pets safe, happy and healthy. The Pawfit 3s pet location and activity tracker puts their minds at ease by enabling them to track their dogs’ location, monitor their activity, set up virtual fences, and use the voice recall to bring back dogs that are out of earshot. Clients can also set fitness goals, get harsh temperature alerts, and more. The device also has a long battery life and is 100% waterproof for hours of carefree adventuring. www.pawfit.com

Curriculum overhaul prepares future vets To better prepare future veterinary graduates for lifelong learning, Penn Vet has significantly revised its curriculum. Even though the former curriculum was cutting edge at the time, it had become outdated and couldn't keep up with modern requirements. Basic scientific discoveries and their application to clinical therapies occur exponentially more quickly now than they did 50 years ago.

The redesigned curriculum will bring fresh changes to how veterinary medicine is taught at the school. Protracted, compartmentalized instructional lectures on essential knowledge are now gone. Shorter and more integrated learning units, including lectures and practical exercises on animal health and illness, as well as independent and critical scientific thinking, have taken their place.

The new curriculum uses a holistic approach to each animal system as a powerful way of learning. Previously, anatomy, histology, and physiology were taught separately; now, students gain knowledge of the anatomical descriptions of certain systems, as well as the associated histology and physiology and the connections between them.

This greatly aids pupils in comprehending how an entire system functions. https://penntoday.upenn.edu/news/penn-vet-educating-21stcentury-veterinarian

New drug for canine osteoarthritis According to recent studies, osteoarthritis is the most common cause of lameness in dogs over a year old. Librela (bedinvetmab injection) was recently given the go-ahead by the FDA for the treatment of canine OA pain, with injectable monoclonal antibody (mAb) therapy expected to become accessible later this year. The drug reduces canine OA pain by targeting nerve growth factor (NGF), and works similarly to naturally-occurring antibodies. www.fda.gov/animal-veterinary/cvm-updates/fda-approves-first-monoclonalantibody-dogs-osteoarthritis-pain

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Osteoarthritis and other musculoskeletal disorders such as ligament injuries, fractures and more, are common in companion animals. Laser therapy, also known as photobiomodulation (PBM) therapy, is becoming an alternative treatment option of choice for these patients.

PBM Therapy for Musculoskeletal Disorders in Companion Animals

BY LISA MILLER, DVM, CCRT, CVA

A growing number of pet owners and veterinary practitioners are seeking adjunct or alternative approaches for treating chronic painful conditions, including osteoarthritis and other musculoskeletal disorders, in companion animals.

Laser therapy, or photobiomodulation (PBM) therapy, is fast becoming a best practice modality for a variety of these conditions. Though PBM may be utilized as a standalone therapy — and in some cases, the results may be better than treatment with NSAIDs alone1 — it is better appreciated as part of a multimodal approach for comprehensive pain management and rehabilitation in veterinary patients.

The basic photochemical processes initiated by PBM on a cellular level have been described in previous articles; however, quite a few overarching short and long-term analgesic and anti-inflammatory physiological effects, as well as effects on modulating cell behaviors and enhancing tissue repair, make this modality particularly useful for treating musculoskeletal conditions.2

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DOSIMETRY FOR MUSCULOSKELETAL CONDITIONS AND GENERAL TREATMENT RECOMMENDATIONS Dosing PBM for musculoskeletal conditions is usually described in terms of the recommended fluence, or energy density (joules/cm2), administered to a specific anatomical area measured in square centimeters and applied at the skin’s surface. It is possible to achieve the same energy density in a number of different ways by adjusting either treatment time or power in watts (1 W = 1 J/s), depending on the laser device.

Treating at higher power allows the same amount of energy (in joules) to be delivered in a shorter time frame than at lower power, although this may not be the only benefit when it comes to deeper tissue conditions. A recent review study looking at efficacious in vitro and in vivo PBM research concluded that the parameters of most importance in PBM treatment success

are both irradiance (mW/cm2) and fluence (J/cm2)13. A minimum threshold of irradiance at the target tissue must be met in order to appreciate a therapeutic result, while a large percentage of light is lost in transmission from the surface of the skin to deeper tissues. This means the fluence and irradiance at the skin’s surface required to produce a therapeutic effect is higher for so-called “deep tissue” conditions, including musculoskeletal disorders, than for “superficial conditions” such as wounds.

One of the conclusions of this review study was that ineffective in vivo results are more likely due to underdosing. As always, in order to optimize treatment, further studies should be encouraged using consistent reported parameters to truly examine the factors necessary for real world clinical efficacy.

In the veterinary literature for orthopedic conditions, positive outcomes have been reported for a general range of fluences between 5-20 J/cm2 based on the size of the patient and the area and depth of tissues being treated.1,14-17 The power (W) used to achieve the desired fluence should be lower for smaller patients, for areas with increased sensitivity, and when the target tissues are not as deep. Software protocols in various laser units differ from company to company, and the laser operator should speak with the PBM being applied on the carpus. manufacturer of their device and have a complete understanding of the dose used for each condition and size area intended for treatment within these protocols. While convenient, however, these software protocols should not replace a thorough understanding of dosimetry, and they should always be checked against the practitioner’s calculations to ensure an effective treatment dose is being delivered.

in soft tissue. A pain examination should be repeated regularly for all patients to evaluate progress, ensure all painful areas are treated, and follow up with any additional diagnostics or other therapies if needed.

If possible, when treating musculoskeletal conditions, the laser treatment head/hand piece should be held in contact with the skin over the affected area, and moved so the entire area is treated evenly. Using contact and applying pressure to the skin creates tissue blanching (dispersal of blood from the dermis and fat tissues)19 and minimizes light losses (which can be significant) due to reflection off the skin.20,13 Some patients with pain may be hypersensitive to physical touch, especially over bony prominences (e.g. elbow and stifle). In these patients, or in those with open wounds or surgical incisions, a non-contact treatment method may be necessary in certain areas. If the patient has a soft bandage or other covering in a postsurgical situation, PBM should be applied during bandage changes since it cannot be performed through bandage, splint, or casting materials.

Most importantly, based on the Principle of Photochemical Activation, photons must reach and be absorbed at the site of “injury” for PBM to be effective.18 Knowing where to treat is essential. All veterinary patients should undergo a thorough orthopedic and pain examination, not only to treat the correct sources of pain, but also to identify any compensatory changes IVC Summer 2023

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A Short Review of PBM Photo courte sy of B ridge A nimal R eferral Cente r

When initiating a course of PBM therapy, the patient may be very stiff and painful during the first few visits. It is best to first address the animal’s discomfort while allowing them to lie or sit on a padded surface, and treat the affected limb(s) in a passive position that allows access to the structures involved. During subsequent treatment sessions, while administering laser treatment over a joint area, the limb may be gently placed through its passive range of motion, if possible and desired.

As a brief general review, PBM uses wavelengths in the red and infrared range to activate cytochrome c oxidase and increase mitochondrial electron transport, inducing a cascade of events leading to an increase in adenosine triPBM therapy for abdominal organs. phosphate (ATP).

This produces beneficial reactive oxygen species (ROS) and nitric oxide, affecting healing and stimulating collagen production via the upregulation of specific substrates and cytokines (including EGF and TGFβ), and the downregulation of others (e.g. interleukin IL-6, IL-8, and IL1)3-8 in treated tissues.

PBM modulates the inflammatory process, and while this is a large part of why it is beneficial for pain relief, it also reduces the sensitization of injured or inflamed peripheral neurons, the dorsal root ganglion (DRG) and spinal cord via modulation of ion channels.

In some situations, it also inhibits Aδ and C fiber transmission.9-12

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1

For any acute or chronic pain condition, an “induction phase” of initial, more frequent treatment sessions is recommended (usually daily to every other day until significant improvement is noted). Following this, the patient can enter a “transition phase” during which treatments are decreased to twice weekly, then once weekly, and so on as the patient improves until the condition resolves (e.g. a soft tissue injury). Alternatively, a “maintenance phase” of treatments is established if the condition is expected to be ongoing (e.g. osteoarthritis).

For musculoskeletal conditions, chronic treatment is typically one session every two to six weeks, based on the patient’s response with a goal of minimizing clinical signs.

OSTEOARTHRITIS

Affecting as much as 20% of the U.S. pet population over the age of one year,21 osteoarthritis (OA) is a progressive syndrome involving the interaction of a multitude of factors. Current treatment strategies in veterinary medicine are aimed primarily at controlling pain, improving joint function, and minimizing functional incapacity in patients through various strategies such as proper weight management, nutraceuticals, pharmacologic therapy, rehabilitation therapy, and when necessary, surgical intervention(s).

One of the most common uses of PBM is for the treatment of chronic pain associated with osteoarthritis. Animal and human trials have shown the modulatory effect of PBM therapy on inflammatory markers and other aspects of the inflammatory process in OA. Studies have demonstrated that treating arthritic joints with PBM reduces edema and the influx of leukocytes,22,23 decreases the concentration of IL-1β and IL-6 in synovial fluid,23 and reduces the activity of

metalloproteinases. 24 Human studies have shown that PBMtreated patients experienced a significant reduction in pain and swelling, as well as an increase in joint mobility and function over the short term.25,26 More recently, these findings are also supported in veterinary research. Dogs with naturallyoccurring elbow arthritis treated with PBM using a fluence of 10-20 J/ cm2 experienced a significant reduction in veterinary assigned lameness scores, and pet owner perceived pain scores; as well, their NSAID requirements decreased compared to a control group.16 Similar findings were reported in a PBM-treated group of military working dogs with moderate to severe hip osteoarthritis versus a control group treated with daily meloxicam for three weeks. Not only did the PBM group experience longer periods with better results versus the NSAID control group, but they also had improved range of motion in the joint up to 90+ days with just three weeks of PBM treatment.1 These results offer a possible alternative of PBM treatment in lieu of systemic NSAIDs, especially early in the diagnosis of osteoarthritis.

Patient preparation, techniques for laser administration, dosing information, and recommended frequency are as described above. While radiographic changes are supportive of an OA diagnosis, they may not be consistent with the level of pain displayed by a patient in a particular area. The PBM treatment area(s) prescribed should be primarily based on orthopedic and pain assessment examinations.

2

TENDON AND LIGAMENT INJURIES

With regard to tendon injuries and tendonitis, PBM literature has focused primarily on the exertion of an antiinflammatory effect after injury, with reduction of proinflammatory mediators.27,28 During the healing process, PBM results in improvement in the remodeling of extracellular matrix through activation of MMP-2 and stimulation of collagen synthesis,29 as well as improvement of collagen fibril size,30 fiber organization,31 and neovascularization.32 These same benefits can also apply in more chronic situations of tendinopathy where discontinuous and disorganized collagen fibers, abnormal neovascularization, edema, and impaired function can present in patients with overuse tendon conditions. In a randomized, placebo-controlled, double-blinded clinical trial, human subjects with chronic lateral epicondylitis received eight treatments with either sham or real laser. The lasertreated group demonstrated improvement in handgrip IVC Summer 2023

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Photo

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sy of M

ilton An im

al Hosp ital

PBM for Muscle Tissue PBM provides several benefits for muscle tissue, but the most prevalent PBM therapy being applied on hip. effects reported in current literature involve the prevention of muscle damage after exercise, including delayed onset muscle soreness, and the improvement of muscle performance.41-43 Clinically, there are still many unanswered questions regarding the use of PBM in canine athletes, including the timing of PBM application to muscles (before, during, or after exercise), the ideal dosimetry, and the exact mechanisms of how PBM interacts with muscle tissue to increase sports performance.

When it comes to the ideal timing for treating muscle tissue, the current body of evidence reports two main strategies for the use of PBM to increase muscle performance and exercise recovery in clinical trials.

The first strategy is the use of muscular pre-conditioning. Numerous rodent and human studies have demonstrated that patients treated with PBM in various time frames prior to exercise are able to perform more repetitions in muscle fatigue tests and have improved muscle energy metabolism compared to control groups.44-48 Studies in humans demonstrated the best improvement anywhere from 40 to 60 minutes to three to six hours prior to exercise.43,49-51 The second strategy applies PBM immediately after exercise in order to accelerate muscle recovery and help heal any muscle injuries via a variety of mechanisms. 49,50,52-54 PBM may also be used any time there is muscle swelling or pain from a variety of causes. It has been used to treat experimentally-induced myositis in rats,55 resulting in a significant reduction in the presence of inflammatory cells and in the amount of edema present in tissue after treatment. Additional studies on experimental myopathy showed significant changes in inflammatory biomarkers and infiltrates, oxidative stress, and improved muscle recovery.56 Regarding dosimetry for performance enhancement or muscle recovery and painful conditions, it is logical and in accordance with previous deep tissue veterinary studies to suggest dosimetry similar to that mentioned in the article, along with treating the largest area possible over the main muscles involved in a particular exercise/sport or injury respectively.49,57 36

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strength, enhanced function, and a continuous reduction in pain over a 12-month period, despite return to normal activity.33 The dosing and frequency of treatments is typical for deep tissue musculoskeletal conditions, with consideration given to the location (depth) of the tendon and other associated soft tissues when selecting power parameters. It should be noted that patients experiencing an acute exacerbation of tendonitis or tendinopathy may be very hypersensitive to pressure, temperature, or both, and that adjustments in treatment power (W) or non-contact versus contact may be necessary.

Some studies have examined cranial cruciate ligament injury and the outcome measures following surgical repair and/ or progression of subsequent osteoarthritis in the knee if no surgery was performed. In lab animal models, laser treatments starting immediately after cruciate ligament transection were shown to prevent some features of articular degeneration of the knee, reducing synovium inflammation, cartilage damage, and knee pain.34,35 In 2017, Renwick et al utilized PBM in dogs for three postoperative treatments following TPLO surgery, and found a greater improvement in the gait section of the adjusted Canine Orthopedic Index questionnaire.36 Rogatko and colleagues investigated the use of PBM as a single preoperative treatment for the same surgery, and found a significant difference in peak vertical force (PVF) in the PBMtreated group. As well, a higher percentage of patients were healed at eight weeks post-op.15 Neither of these studies showed significant differences in bone healing; however, this author would refer back to previous comments regarding the increased dosing needed for deep tissue conditions. In the former canine study, doses listed in the appendix section show laser settings that were less than 50% of those recommended for deep tissue conditions (including bone healing), perhaps suggesting that dosimetry, while adequate for some pain relief benefits, was not high enough for other outcomes to be significant.

Treatment with PBM therapy should begin immediately after CCL surgery (if not before) to address pain and inflammation and assist in wound healing. The incisional area and affected superficial soft tissues may be treated with a more superficial target energy density of 4-6 J/cm2, and with a lower power (W) setting than would be used in a deeper tissue dose for the stifle joint itself. Deeper soft tissues of the anatomic region that are painful or bruised, or become edematous after surgery, may be treated with a higher fluence (~10-20 J/cm2, depending on the size of the patient). The laser operator should also consider treating the contralateral stifle, due both to the predisposition of CCL tear on the opposite limb as well as compensatory

weight shifting. The author also recommends examination and possible treatment of the iliopsoas muscle and lower back, since these areas are often also quite painful in these patients.

In acute postoperative situations, treatment should ideally be performed daily, if possible, for the first 24 to 72 hours, then twice to three times weekly, as with other chronic conditions, tapering as the patient improves.

When treating partial or complete CCL tears managed conservatively in some other fashion (bracing, etc.) due to any reason (financial restrictions, co-morbidities, etc.), expectations should be set with the pet owner that the goal of PBM treatment is only for pain management along with reducing inflammation and possibly cartilage damage.

Typical dosing for deep tissue musculoskeletal conditions should be used. Treatment for the stifle should include a relatively large area, incorporating all the important soft tissue structures in the area, including the insertion of the hamstring muscles and the patellar ligament. Ideally, all the above should be performed as part of an appropriate course of physiotherapy, with continuous reassessment and adjustments along the way.

3

FRACTURES AND BONE HEALING

In addition to the analgesic benefits PBM brings to fracture repair cases, there is evidence in the literature that it can be used in situations where bone healing is desired. In a recent review of 25 relevant articles regarding the use of PBM in bone healing, 11 of 13 in vitro studies showed positive results with regard to the acceleration of cell proliferation and differentiation, and all animal studies showed improved bone healing in laser-treated sites.37 A later systematic review found that 75 of 76 studies utilizing PBM showed positive effects on fractures in animal models, including acceleration and stimulation of fracture healing as well as callus maturation.38 Mota and colleagues showed that even though PBM’s effects were more prominent when treatment started during the acute phase of the injury, it still aided the bone consolidation process and favored the physiopathologic mechanisms involved in bone tissue repair when used in the chronic phase as well.39

Further controlled studies in veterinary patients are needed; at this time, however, treatment should ideally be performed immediately IVC Summer 2023

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after stabilization, and then done twice weekly for a total of six to eight treatments.40 A target energy density suitable for deep tissues (10-20 J/cm2) is recommended, scaling up with depth of the soft tissues overlying the area. The entire area to be treated should include not only the soft tissues overlying the fracture itself, but several square centimeters proximal and distal.

Rogatko, et al. (2017) Preoperative low level laser therapy in dogs undergoing tibial plateau levelling osteotomy: A blinded, prospective, randomized clinical trial Vet Comp Orthop Traumatol 2017; 30(01): 4653. DOI: 10.3415/VCOT-15-12-0198

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Looney, A.L., et al. (2018) A randomized blind placebo controlled trial investigating the effects of photobiomodulation therapy (PBMT) on canine elbow osteoarthritis. Can Vet J. 59(9):959–66.

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Barale, L., et al. (2020) Preliminary clinical experience of low-level laser therapy for the treatment of canine osteoarthritis-associated pain: A retrospective investigation on 17 dogs. Open Vet J. 0(1):116–9.

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Calvert, J., and Pitts, J. (1966) Photochemistry. New York: Wiley & Sons.

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In summary, a large body of research in both veterinary and human medicine elucidates the potential beneficial effects of PBM on conditions of the musculoskeletal system.

Further research should be encouraged to uncover additional molecular mechanisms, timing of treatment, and optimal dosing parameters for various conditions. We should not forget this modality when managing veterinary patients for these common conditions.

Chien, S. & Wilhelmi, B.J. (2012) A simplified technique for producing an ischemic wound model. Journal of visualized experiments : JoVE, (63), e3341.

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Piao D., et al. (2018) Flexible nine-channel photodetector probe facilitated intraspinal multisite transcutaneous photobiomodulation therapy dosimetry in cadaver dogs. J Biomed Opt. 23(1), 010503.

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Johnston, S.A. (1997) Osteoarthritis. In: Veterinary Clinics: Small Animal Practice. 27(4): 699–723.

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Bjordal JM, et al. (2006) Photoradiation in acute pain: a systematic review of possible mechanisms of action and clinical effects in randomized placebo-controlled trials. Photomed Laser Surg. 24(2):158–168.

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Pallotta, R.C., et al. (2012) Infrared (810-nm) low-level PBMT on rat experimental knee inflammation.

Lasers in Medical Science. 27(1):71-78.

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Carlos, F.P. et al. (2014) Protective effect of low-level PBMT (LLLT) on acute zymosan-induced arthritis.

Lasers in medical science, 29(2), 757–63.

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Alves, J. C., et al. (2022). A randomized double-blinded controlled trial on the effects of photobiomodulation therapy in dogs with osteoarthritis. American Journal of Veterinary Research. 83(8).

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Pryor, B. & Millis, D.L. (2015) Therapeutic Laser in Veterinary Medicine. In: Veterinary Clinics of North America: Small Animal Practice. 45(1): 45–56.

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Baltzer, A.W.et al. (2016) Positive effects of low level PBMT (LLLT) on Bouchard’s and Heberden's osteoarthritis. Lasers in surgery and medicine. [Epub ahead of print] Fukuda, V.O. et al. (2015) Short term Efficacy of Low Level PBMT in Patients with Knee Osteoarthritis: A Randomized, Placebo-Controlled, Double-Blind Clinical Trial. Revista Brasileira de Ortopedia, 46(5), 526–533.

Passarella, S. et al. (1984) Increase of proton electrochemical potential and ATP synthesis in rat liver mitochondria irradiated in vitro by helium-neon laser. FEBS Lett. 175, 95–99.

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Morimoto, Y., et al. (1992) Effect of low-intensity argon laser irradiation on mitochondrial respiration. Lasers Surg Med. 1992; 15, 191–199.

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Karu TI. The Science of Low Power Laser Therapy. London, UK: Gordon and Breach Science Publishers; 1998.

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Da Ré Guerra, F. et al. (2016) Low-level PBMT modulates pro-inflammatory cytokines after partial tenotomy. Lasers in medical science.

Pires, D. et al. (2011) Low-level PBMT (LLLT; 780 nm) acts differently on mRNA expression of anti- and pro-inflammatory mediators in an experimental model of collagenase-induced tendinitis in rat. Lasers in medical science. 26(1): 85–94.

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Karu TI. Primary and secondary mechanisms of action of visible to near-IR radiation on cells. J Photochem Photobiol B. 1998; 49:1–17.

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Stadler, I., et al. (2001) 830-nm irradiation increases the wound tensile strength in a diabetic murine model.

Lasers Surg Med. 28, 220–226.

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Hamblin, M. R., and Demidova T.N. (2006) Mechanisms of low level light therapy. SPIE Proc. 6140:1–12.

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Holanda, V.M., et al. (2016) Photobiomodulation of the Dorsal Root Ganglion for the Treatment of Low Back Pain: A Pilot Study. Lasers Surg Med. 48(7):653-9.

Guerra, F.D.R. et al. (2013) LLLT improves tendon healing through increase of MMP activity and collagen synthesis. Lasers in medical science. 28(5): 1281–8.

Ng, G.Y.F. et al. (2004) Comparison of single and multiple applications of GaAlAs laser on rat medial collateral ligament repair. Lasers in surgery and medicine. 34(3): 285–9.

Oliveira, F.S. et al. (2009) Effect of low level PBMT (830 nm) with different therapy regimes on the process of tissue repair in partial lesion calcaneous tendon. Lasers in surgery and medicine. 41(4): 271–6.

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Tsuchiya, D., et al. (1994) Laser irradiation abates neuronal responses to nocioceptive stimulation of rat paw skin. Brain Res Bull. 43:369–374.

Salate, A.C.B. et al. (2005) Effect of In-Ga-Al-P diode laser irradiation on angiogenesis in partial ruptures of Achilles tendon in rats. Photomedicine and laser surgery. 23(5): 470–5.

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Chow, R., and Armati P. (2004) Effects of 830nm laser on cultured rat dorsal root ganglia: Implications for the analgesic effects of laser. Lasers Surg Med.(Suppl. 6):5.

Roberts, D.B., et al. (2013) The effectiveness of therapeutic class IV (10 W) laser treatment for epicondylitis.

Lasers in surgery and medicine. 45(5): 311–7.

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Holanda, V., et al. (2017) The Mechanistic Basis for Photobiomodulation Therapy of Neuropathic Pain by Near Infrared Laser Light. Lasers Surg Med. 49(5):516-524.

Bublitz, C. et al. (2014) Low-level PBMT prevents degenerative morphological changes in an experimental model of anterior cruciate ligament transection in rats. Lasers in medical science. 29(5): 1669–78.

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Zein, R., et al. (2018) Review of light parameters and photobiomodulation efficacy: dive into complexity. J Biomed Opt. 23(12):1-17. doi: 10.1117/1.JBO.23.12.120901.

Wang, P. et al. (2014) Effects of low-level PBMT on joint pain, synovitis, anabolic, and catabolic factors in a progressive osteoarthritis rabbit model. Lasers in medical science. 29(6): 1875–85.

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Gaynor J., (2015) Energy Modalities, Therapeutic Laser and Pulsed Electromagnetic Field Therapy. In: Handbook of Veterinary Pain Management. (eds J. Gaynor & W. Muir) 3rd edn, pp. 357-362. Mosby, St. Louis, Missouri.

Renwick, S.M.,et al. (2018) Influence of class IV laser therapy on the outcomes of tibial plateau leveling osteotomy in dogs. Veterinary Surgery. 47: 507- 515. https://doi.org/10.1111/vsu.12794

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Ebrahimi, T. et al. (2012) The influence of low-intensity PBMT on bone healing. Journal of dentistry (Tehran, Iran). 9(4): 238–48.

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Bayat, M., et al. (2017) Comparison of effects of LLLT and LIPUS on fracture healing in animal models and patients: A systematic review. Progress in Biophysics and Molecular Biology. 132: 322, https://doi. org/10.1016/j.pbiomolbio.2017.07.004.

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Mota, F.C.D. et al. (2013) Low-power PBMT for repairing acute and chronic-phase bone lesions.

Research in Veterinary Science. 94(1): 105–110.

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Godbold, J. & Arza R. (2011) Reference Guide to Common Conditions for CTC, CTS, p. 10. LiteCure, LLC., Newark, DE.

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Ferraresi, C., et al. (2016) Photobiomodulation in human muscle tissue: an advantage in sports performance? J Biophotonics. 9(11-12):1273–99. doi: 10.1002/jbio.201600176.

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Hamblin, M.R. (2017) Mechanisms and applications of the anti-inflammatory effects of photobiomodulation. AIMS Biophys. 4(3):337–61. doi: 10.3934/biophy.2017.3.337.

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Ferraresi, C., et al. (2015) Time response of increases in ATP and muscle resistance to fatigue after low-level laser (light) therapy (LLLT) in mice. Lasers Med Sci. 30(4):1259–67. doi: 10.1007/s10103015-1723-8.

From the AVH The Academy of Veterinary Homeopathy is comprised of veterinarians who share a common desire to restore true health to their patients through the use of homeopathic treatment. Members of the Academy are dedicated to understanding and preserving the principles of Classical Homeopathy.

VETERINARY GASTROENTEROLOGY AND HOMEOPATHY

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Leal, E.C.P., et al. (2010) Effects of Low-Level Laser Therapy (LLLT) in the Development of ExerciseInduced Skeletal Muscle Fatigue and Changes in Biochemical Markers Related to Postexercise Recovery.

J Orthop Sport Phys Ther. 40(8):524–32. doi: 10.2519/jospt.2010.3294.

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De Marchi, T. (2012) Low-level laser therapy (LLLT) in human progressive-intensity running: effects on exercise performance, skeletal muscle status, and oxidative stress. Lasers Med Sci. 27(1):231–6. doi: 10.1007/s10103-011-0955-5.2006.24.377.

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Lopes-Martins, R. A. B., et al. (2006) Effect of low-level laser (Ga-Al-As 655 nm) on skeletal muscle fatigue induced by electrical stimulation in rats. J Appl Physiol. 101(1):283–8. doi: 10.1152/ japplphysiol.01318.2005.

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Vieira, W.H., et al. (2012) Effects of low-level laser therapy (808 nm) on isokinetic muscle performance of young women submitted to endurance training: a randomized controlled clinical trial. Lasers Med Sci. 27(2):497–504. doi: 10.1007/s10103-011-0984-0.

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Levine, D., et al. (2015) Effects of Laser On Endurance of the Rotator Cuff Muscles. Lasers in Surg Med. 47(S26) 44-45. doi: 10.1002/lsm.22351.

Good food and nutrition are paramount to our veterinary patients’ health. Chronic disease can hinder good nutrition, and a poor diet can be an obstacle to homeopathy. A mistuned vital force cannot use even the most excellent food very well until the correct remedy is given.

The imbalance caused by chronic disease impedes normal digestion.

As a truly homeopathic remedy restores balance, digestion improves, resulting in enhanced tissue health and organ function.

Here are some key acute remedies to consider for gastrointestinal symptoms: • A rsenicum album — keynote is restlessness; vomiting and diarrhea (foul odor); often bloody stool; craves frequent small sips of cold water; food poisoning or garbage ingestion.

• Ipecac — nausea and vomiting at the mere smell of food; no thirst; bloody stool; gets diarrhea easily.

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Ferraresi, C., et al. (2013) Low-level laser (light) therapy (LLLT) on muscle tissue: performance, fatigue and repair benefited by the power of light. Photon. Lasers Med. 1(4), 267–286

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Borsa, P.A., et al. (2013) Does phototherapy enhance skeletal muscle contractile function and postexercise recovery? A systematic review. J Athl Train. 48(1):57–67. 10.4085/1062-6050-48.1.12.

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Ferraresi, C., et al. (2015) Light-emitting diode therapy (LEDT) before matches prevents increase in creatine kinase with a light dose response in volleyball players. Lasers Med. Sci. 30(4), 1281–1287. [doi: 10.1007/s10103-015-1728-3].

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Douris, P., et al. (2006) Effect of Phototherapy on Delayed Onset Muscle Soreness. Photomed Laser Surg. 24(3):377–82. doi: 10.1089/pho.

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Sussai, D.A., et al. (2010) Low-level laser therapy attenuates creatine kinase levels and apoptosis during forced swimming in rats. Lasers Med Sci. 25(1):115–20. doi: 10.1007/s10103-009-0697-9.

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• N ux vomica — a top remedy for many gastrointestinal issues; treats bad effects of drugs and chemicals; overeating; seizure prone; frequent urge to stool; irritable/oversensitive/grumpy when ill.

The following are two important remedies for chronic disease, often helpful with gastrointestinal symptoms: • C alcarea carbonica — a very important nutritional remedy; keynote is poor growth/development; stunted appearance; distorted breeds; delayed dentition/retained deciduous teeth; orthopedic problems/ arthritis.

• S ulphur — good appetite but thin; slow, lazy; unkempt appearance; looks old; anus and mouth red/inflamed; bloody discharges with odors; swollen belly; thin legs; liver problems.

Since it is hard to distinguish poor nutrition from chronic disease, try the following approach:

Dos Reis, F.A., et al. (2014) Effects of pre- or post-exercise low-level laser therapy (830 nm) on skeletal muscle fatigue and biochemical markers of recovery in humans: double-blind placebo-controlled trial.

Photomed. Laser Surg. 32(2), 106–112 (2014) [doi: 10.1089/pho.2013.3617].

1. I mprove diet as much as possible.

Carvalho, A.F.M.D., et al. (2015) The low-level laser on acute myositis in rats. Acta Cirurgica Brasileira, 30(12): 806-811.

3. E liminate obvious drugs and chemicals from the picture if possible

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2. T reat with the best homeopathic remedy for the total symptom picture (no suppressive treatments).

Servetto, N. et al. (2010) Evaluation of inflammatory biomarkers associated with oxidative stress and histological assessment of low-level PBMT in experimental myopathy. Lasers in surgery and medicine.

42(6): 577–83.

You can find a veterinary homeopath at theAVH.org.

Ferraresi, C., et al. (2011) Effects of low level laser therapy (808 nm) on physical strength training in humans. Lasers Med. Sci. 26(3), 349–358 (2011) [doi:10.1007/s10103-010-0855-0].

Submitted by Todd Cooney, DVM, CVH

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Creating a sustainable marketing strategy for your veterinary practice

BY MEGAN KELLY, BVSC, CCRP

As a veterinary professional, you also need to be a marketer — and learn how to measure your marketing efforts for optimal success.

THE ABSOLUTE BASICS OF MARKETING

1

onsistency is key: To see results, you really C have to be consistent with your marketing efforts.

Blowing hot and cold will not bring in the desired traction. Consistency is the key to success — as in all things.

Don’t spread yourself too thin: One of the biggest Are you familiar with John Wanamaker’s classic saying: “Half my money I spend on marketing is wasted — I just don’t know which half”? If you aren’t, you’re probably not yet aware of the importance of measuring your marketing efforts. In this article, I explain why this is so crucial, and share some tips on how to market — and measure your marketing — effectively.

As a self-taught marketer, I’ve immersed myself in the topic, devouring hundreds of blogs and books on marketing every month. While my background is in veterinary science, my colleagues know me as someone who gets genuinely excited when discussing anything about marketing. Unfortunately, for many people, marketing is a necessary evil that lacks excitement or strategic planning. Instead, they just mimic what other businesses are doing or copy the strategies of big brands in the hopes of achieving similar results. That really is not ideal; to be genuinely effective, your marketing needs to be unique, memorable, consistent, planned, flexible — and measured.

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mistakes businesses make is trying to be on every social media platform. You simply spread yourself too thin and dilute your efforts. Instead, choose two platforms, one primary and one secondary, and focus on creating engaging content on those.

Measure your marketing efforts: Measuring is

crucial for determining what works and what does not. Once you figure out what is effective, you can focus your resources on the strategies that are actually driving results.

Stay adaptable: The environment in which we work

4

is constantly changing, and it’s essential that our marketing stays relevant. New businesses pop up, the desires of customers change, your team takes on a new skill or facility, the profession develops, and social media algorithms can change at the drop of a hat. You need a marketing strategy that is planned but flexible.

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Be unique: Don't copy what everyone else is doing.

Try to stand out from the crowd. Be unique and remarkable! Being different is what gets you noticed and remembered — and it makes marketing so much more fun.

AIM FOR A SUSTAINABLE MARKETING STRATEGY A sustainable marketing strategy means a consistent and measurable strategy. Define what consistency means for your practice — this will depend on factors such as the size of your team, your available resources, and your overall goals. Decide to do a few things well rather than trying to a lot of things more or less okay.

• O ne of the best ways to ensure consistency is to plan. Create a quarterly and monthly social media content plan and run one or two themed campaigns each month. Then regularly assess your tactics to see what is working and what is not. By consistently reviewing your strategies and measuring your results, you can refine your marketing plan over time to maximize impact. This approach will help you create a sustainable marketing strategy that drives long-term growth for your practice.

• N ext, decide what you will measure — i.e. your key performance indicators (KPIs). I used to measure all the wrong things in my business. It was only when I got a Google Analytics specialist to look into my website

SISTENCY CON IS

KE Y

traffic that I realized 4,000 of my site’s monthly visits were for a keyword in a blog that was completely unrelated to what I did. I was getting a lot of traffic, but most of it was the wrong traffic — which is why my conversion rate (traffic to engagement) was so low.

Measuring the effectiveness of your marketing efforts is crucial to understand what is working and what is not. For this, you need to know what KPIs to measure.

KPIS TO MEASURE onversion rate: This measures the percentage of C visitors to your website who take a desired action, such as filling out a form to book a consult or, if you have an online shop, making a purchase. A high conversion rate indicates your marketing is effective at driving user engagement and sales. The average is a 5% conversion rate. You can see your conversion rate by accessing your Google Analytics data. You might need to create some tracking goals. An SEO or Google Analytics specialist can help you with this.

Return on investment (ROI): This is a measure of revenue generated from your marketing efforts in relation to the amount spent. If you made $1,000 from a $100 effort, your return on investment would be 0.9, or 90%. A high ROI means your marketing is effective. I am often asked: “What should I budget for my marketing?” For most practices, assume 6% to 14% of income. If you’re getting a high return on your investment, the marketing budget really does not matter; your only limit will be how many clients you can actually treat in a day.

However, money is not the only consideration when it comes to ROI. Time is precious, too. The time you or a team member spends on marketing must justify the return.

Evaluate how much time you spend on marketing, especially social media marketing. Even if those beautifully curated Instagram stories and posts are generating results, is the time you spend on them worth it? It is very easy to get sucked down the social media rabbit hole.

Think about the time spent creating posts that are IVC Summer 2023

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seen by only 1% to 5% of your followers, depending on the algorithm. Most of these will disappear after 24 hours. Spend more time creating evergreen content you can use over again.

Repurposing social media content is always recommended. You can share a video to different platforms, transcribe the audio and use the text for a blog. Also be sure to reshare social media posts that did well, especially ones that share valuable content.

To enhance ROI, have a clear goal for every post you create and schedule — perhaps to increase brand awareness (a general goal); drive engagement (a specific goal); or generate traffic (a focused goal).

You might want to educate or entertain, showcase a specific service or cluster of services, or simply maintain relationships with established clients. Each of these is quite different. Perhaps include a variety of types each month. Then measure whether each post has achieved its goal.

SOME KEY TAKEAWAYS • B e consistent.

• F ocus on one or two social media platforms.

• P ost unique and interesting content that reflects your business and your personality.

• H ave a plan and stick to it, devising a couple of key campaigns for each quarter, with timed content released each month and each week.

• B e flexible — adjust marketing messages as conditions change.

• M easure your KPIs. • C ombine excellent service with consistent, measured marketing for optimum results.

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Customer acquisition cost (CAC): This measures the cost of acquiring a new customer. A lower CAC means your marketing is effective; it is generating new customers at a low cost. Knowing how much you spend gives you a way to track whether or not this amount is increasing. The lower the CAC, the better. ngagement metrics: A higher engagement rate E indicates your marketing is effective, since it is generating interest and building a community around your brand. In social media marketing, engagement metrics are the most important KPI. Beware of vanity metrics. These look impressive on the surface but do not provide any meaningful insights or contribute to the achievement of business goals.

They are often easy to measure and are used to inflate the perceived success of a marketing campaign — e.g. likes, shares, and followers on social media, website traffic and page views. These metrics may make a practice look good, but they do not necessarily indicate success or a positive return on investment.

Focusing on vanity metrics can distract practices from measuring and improving metrics that are truly important for achieving their objectives. My advice is, don’t get caught up in vanity metrics; focus on metrics that provide real insight into how your marketing is translating into engagement and customer acquisition.

Brand awareness: This measures the level of recognition and recall of your brand among your target audience. A high brand awareness indicates your marketing is effective at creating a memorable and recognizable brand.

Customer retention: Measuring the percentage of customers who return to make repeat purchases is important. This is an area on which we really need to focus. Ultimately, it has little to do with marketing, and far more to do with customer experience. Spend time with each customer, show genuine interest in each case, and give a great client experience — that’s how you win repeat customers. When consistent marketing is backed up by a great experience, you have a winning combination. Remember as well that it is cheaper to retain a customer than to acquire a new one.

YOU’RE A PROFESSIONAL AND A MARKETER Yes, you’re a professional — but a successful professional in an age of social media and personal branding is a marketer too. Your key to success is consistent marketing sustained over time, regularly measured and adjusted.

If you want your practice to grow, you are going to have to learn to love marketing, or at least embrace it. It really can be fun — and the better you are at it, the stronger the community around you becomes and the more animals you’ll reach. So let your marketing skills grow alongside your ever-evolving professional skills.

Twice a month Dr. Kelly curates hundreds of social media trends and updates in her Social Media Buddy free email newsletter. If you’re interested, you can sign up at www.drmegankelly.com/ social-media-buddy.

From the AHVMA The mindful leader elevating the veterinary profession through innovation, education and advocacy of integrative medicine.

ANNUAL CONFERENCE IN OCTOBER Register now for the 2023 AHVMA Annual Conference and Exhibition, taking place in San Diego, California from October 7 to 10. Expect over 100 hours of complementary and integrative CE sessions, hands-on labs, and special events.

Over the past six years, AHVMA has applied for and received AAVSB RACE approval for over 80 hours of continuing education.

This year’s keynote address, “NeuroDiversity in a NeuroTypical World” by Laurie McCauley, will have us thinking differently and learning more about ourselves and how we see the people around us.

You can expect to find lectures on cannabis and mushrooms, homeopathy, rehabilitation, TCVM, ozone, and so much more! Look for our introductory talks on various holistic modalities. Not only will you be exposed to a wealth of knowledge; you will also acquire information on how to integrate these modalities into your practice. We encourage students from AVMA-accredited veterinary schools to attend these lectures as a way to broaden their veterinary education.

AHVMA RETREAT This year’s post-conference retreat takes place on October 10, immediately following the conference, and will be held at the Santa Clara Recreation Center on the water in Mission Bay, from 2 pm to 7 pm. More details regarding registration for the retreat will be updated on the AHVMA website.

INTERESTED IN SPEAKING AT THE 2024 CONFERENCE? The Conference Program Committee is still taking speaker submissions for next year’s conference. AHVMA is excited to announce that the Call-for-Papers Portal has been upgraded and is now more user-friendly! For consideration, submit your proposal(s) via the online portal no later than the November 1 deadline. Those selected will be contacted by the AHVMA CPC. ahvma.org Submitted by Melissa Kellagher, AHVMA Director of Operations

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HOW

MBRT

HELPED A DOG RECOVER FROM AN ANTIBIOTICRESISTANT INFECTION

BY MARGO ROMAN, DVM, CVA, COT, CPT, FAAO & DAMAGED AUTONOMIC NERVES Used in combination with ozone therapy and other modalities, Microbiome Restorative Therapy (MBRT) led to full recovery for an injured dog that developed an antibiotic-resistant urinary tract infection, along with sustained damage to his autonomic nerves.

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As a veterinarian, my primary aim is to help animals improve and increase their health. Backed by scientific knowledge and 45 years of experience, I’ve been using Microbiome Restorative Therapy (MBRT) for 12 years, and have found it to be the most effective way to treat animals and strengthen their immune systems. In this article, I’ll highlight how MBRT along with other therapies helped an injured dog with an antibiotic resistant infection, as well as damaged autonomic nerves, make a full recovery.

In recent years, MBRT has achieved prevalence for helping patients recover optimal gastrointestinal health. It involves transplanting gut micro-organisms from the fecal matter of a fit and healthy animal into a diseased one. Used in combination with ozone therapy, MBRT is even more effective, because ozone helps remove disease-causing resistant biofilms from surfaces in the body, and also reduces pain. This combination therapy establishes the foundation for rebooting a patient’s immune system.

EXPLAINING MICROBIOME RESTORATIVE THERAPY (MBRT)

CASE STUDY — TITAN

The gut of a living organism, such as a dog or human, is inhabited by a unique and varied collection of thousands of beneficial microbes, known as the microbiome. This community of microbes contributes to host digestion, defends the animal or person against pathogens, trains the immune system and central nervous system, helps with behavior issues (e.g. anxiety and depression), and establishes direct or indirect connections to physiological functions.

Serotonin, one of the important neurotransmitters, is mostly produced in the intestine and illustrates the gutbrain axis concept.

The microbiome contributes to 75% of immune system development. It helps restore the precursors of neurotransmitters, and promotes food absorption. However, the gut microbiome can be damaged by many things, including antibiotics, NSAIDs, pesticides, herbicides, preservatives, and other contaminants, resulting in a patient with a weakened ability to fight infections.

A 14-month-old male Rottweiler named Titan was hit by a car and suffered a severely comminuted fracture of vertebra L7 with luxation of S1, which was surgically repaired. After two months of university care, he still had no anal tone and was experiencing excoriating diarrhea and a bleeding ulcerated tail. He was only able to walk 20 feet. He also had an atonic bladder with a non-responsive antibiotic-resistant urinary tract infection that continued to smell like dead fish even after two months of chloramphenicol treatment. Titan was being given two tablets of chloramphenicol (1 g/tablet) by mouth three times a day, with food or shortly after a meal. Side effects from the chloramphenicol included vomiting, anorexia, diarrhea and severe bone marrow suppression. After nine weeks on the medication, Titan was still not responding and was passing foul brown urine. Many veterinarians told his owners to euthanize him. He was fearful and insecure, and had a hard time balancing when in the exam room, so he leaned on the walls and smeared blood and diarrhea all over them.

At Titanʼs first integrative exam, we saw the same resistant bacterial strain that was initially present. His bladder and gut needed a reboot with balanced species. We took DNA culture from his urine and evaluated him for regular aerobic and anaerobic infection. Titanʼs urinary infection was so hazardous that when the familyʼs baby boy came in contact with the infected dogʼs urine, he was exposed to the chloramphenicol, developed an allergic reaction all over his body, and was admitted to the emergency room.

To treat Titan for this painful and debilitating situation, a combination of therapies was used, including ozone, aquapuncture, homeopathy, laser, UVBI therapy, and MBRT herbs and nutraceuticals.

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DOSES AND TREATMENT PROCEDURE

In MBRT, the microbiome is obtained from healthy donors. We use the term “Microbiome Restorative Therapy” in place of “Fecal Microbiota Transplantation” or FMT because our aim is to restore the microbiome and not just transplant the poop. The donor dogs from Touch of Life Microbiome (www.mbrt.life) are from six generations that have not been given antibiotics, flea and tick products, or NSAIDs. They are given fresh homemade raw organic food and natural well water, and live in virgin woods in a green sustainable non-toxic home, where they are thriving. These donors are predominately plant-based dogs and are holistically reared. The small amount of meat they eat — less than 7% — is mostly wild venison from hunting season.

When combining ozone therapy with microbiome therapy, the ozone gas is used to remove the resistant biofilm in the gut and allow the animal to defecate, providing a more unpolluted intestinal surface. After this, we inoculate the same region of the intestines with a slurry of fecal from a healthy biodiverse donor.

Animals are standing during the procedure, and anesthesia is not necessary. We give some time for the new microbiome to mesh with some of the old, but also start all new colonization.

Our preference is not to allow the pet to defecate until after a parasympathetic sleep period. We intensify the process by nurturing the gut flora with nutraceuticals that support digestion and communication between the microbes, as well as help repair leaky gut and other damages.

Each diseased individual is provided with a different protocol depending on what support they need. Ozone therapy also enhances the body’s innate ability to heal. It should be remembered that adding medications, antibiotics and pharmaceuticals can affect the symbiotic balance that helps in the healing process.

It’s important to try to do things more naturally, with homeopathy, herbs and nutraceuticals as your first choice. Allowing the microbiome to communicate with other body systems is the new future of medicine.

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• U VBI therapy was administered with a dose rate of 50 cc ozonated saline, 5 cc blood, and 1 cc hypertonic Biocean. We used Hemealumen, which utilizes UVA, UVC, amber, blue, red and green light for photobiomodulation.

• S ubcutaneous ozone saline at a saturation of about 37 ug/ml was flooded over the surface of the pelvis at the rate of approximately 150 cc, and distributed from L3 to the base of the tail.

• A quapuncture with B12 was given at BL 60, 40 23, 57 and all along the sacrum and lumbar; SP 6, ST 36, 25; Bau Hui, at the tip of tail, under tail and base of tail; LI 11, GV 20 and GB 20. When injecting the aquapuncture, the fascia was so dense and constricted it was like putting needles through beef jerky instead of muscle.

• A fter the aquapuncture, the MultiLaser product for neuropathy from Advanced Light Devices was administered for 20 minutes over Titan’s back. The infection frequency was also used over the bladder. The LED laser of the multi-lumen with rife frequencies was used.

• W ith the drugs discontinued (we stopped all antibiotics, pain medications and Urecholine), we started oral microbiome supported with nutraceuticals to restore the oral and gut microbiome.

• A catheter was placed within the bladder, and UA and cultures were extracted for examination; 60 cc of ozonated saline was given as a flush and removed with the addition of 60 cc of O3 gas at 41 ug/ml.

• A n additional 5 cc of vitamin C, 2 cc of vitamin B complex, and 1 cc of vitamins A and D were added into two separate 30 cc pockets of O3 saline.

• A fter Titan’s second treatment, his tail wagged for the first time since his injury. A MBRT was added to the third treatment. He had been taking oral MBRT since his initial visit. Titan miraculously recovered on Day 6, regaining his anal tone and passing normal stools. He was able to urinate on his own and the urine was clear.

All of Titan’s autonomic nerve function was restored, and he was able to walk normally for two miles. Even more importantly, his happy playful disposition returned.

After four weeks, however, Titan suddenly had blood in his urine. His urinalysis showed a high pH of 9 and some amorphous phosphate crystals.

• W e considered doing a urine biome transplant. This had never been done before in the literature and his owner, a nursing student, was excited to be the first. A donor in Vienna was selected; at eight years of age, she has been a microbiome fecal donor her whole life but had never donated urine for a urine transplant. The sample was obtained first thing in the morning as a free stream in a sterile cup and was kept refrigerated until we used it.

• T itan was given UVBI therapy with Hemealumen, with increased volume of 80 cc, 8 cc of blood and 2 cc of hypertonic Biocean. As well, he was given sq ozone saline 120 cc over his pelvis.

• A fter that, ozone gas only at 60 cc was introduced within the bladder at 41 ug/ml to remove any biofilm. No saline flush was done; the ozone gas stayed for 15 minutes to allow it to reduce the biofilm. Ozone also increases nitrous oxide (NO) in the tissue, and flooding the back and bladder with ozone increased NO in these areas.

REFERENCES

Pilla R, Suchodolski JS. (2020). The role of the canine gut microbiome and metabolome in health and gastrointestinal disease. Frontiers in veterinary science, 6, 498.

Bonforte G, Bellasi A, Riva H, Ferradini M, Arrighi E, Groppi G, Franzini M. (2013).

Ozone therapy: a potential adjunct approach to lower urinary tract infection? A case series report. Giornale Italiano di Nefrologia: Organo Ufficiale Della Societa Italiana di Nefrologia, 30(4), gin-30.

Dai T, Vrahas MS, Murray CK, Hamblin MR. (2012). Ultraviolet C irradiation: an alternative antimicrobial approach to localized infections? Expert review of antiinfective therapy, 10(2), 185-195.

Elvis AM, Ekta JS. (2011). Ozone therapy: A clinical review. Journal of natural science, biology, and medicine, 2(1), 66.

Hamblin MR. (2017). Mechanisms and applications of the anti-inflammatory effects of photobiomodulation. AIMS biophysics, 4(3), 337.

Hamblin MR. (2017). Ultraviolet irradiation of blood: “the cure that time forgot?”

Ultraviolet Light in Human Health, Diseases and Environment, 295-309.

Masan J, Sramka M, Rabarova D. (2021). The possibilities of using the effects of ozone therapy in neurology. Neuroendocrinol. Lett, 42, 13-21.

Roman M. (2015). Micro-biome restorative therapy: successful treatment of dogs and cats with fecal transplants. J Am Holistic Veterinary Med Assoc, 38, 8-12.

Tashiro H, Shore SA. (2021). The gut microbiome and ozone-induced airway hyperresponsiveness. Mechanisms and therapeutic prospects. American journal of respiratory cell and molecular biology, 64(3), 283-291.

Yong E. 2016. I Contain Multitudes: The microbes within us and a grander view of life.

Random House.

• N ext, 30 cc of the donorʼs urine was injected in the same catheter. The catheter was pulled, the urine was massaged around the bladder, and the patient was prescribed to not urinate for as long as possible, in this case about six hours.

Titan walked five miles the following day, and an additional three miles after the Urobiome transplant. He has not had any more issues with his bladder for more than a year. He runs and plays like any young dog, and you would never think he’d had such a serious injury.

The utilization of these modalities helped reduce Titan’s infection by restoring the microbiome, while also decreasing inflammation and trauma to the tissue. As well, neurotransmitters originating from the microbiome helped with the recovery of the nerves damaged in the accident. Titan’s owners got their happy dog back, and to date he’s still doing well and living a normal life. I would like to thank his owner, who supported and allowed this therapy for the betterment of their dog’s life and well-being.

Having done 30,000 MBRT cases, in combination with ozone therapy, I can say that this approach has displayed incredible results and a major shift in outcomes.

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TCVM APPROACH TO FELINE COLITIS

By identifying the patterns of disharmony contributing to the development of feline

colitis, the TCVM practitioner can use several

modalities to treat the problem and bring the cat’s body back into balance.

BY CHRIS BESSENT, DVM, MSOM, DIPL. OM, LAC

Feline colitis is an inflammation of the colon in cats, causing diarrhea, abdominal discomfort, changes in appetite, and other symptoms. Traditional Chinese Veterinary Medicine (TCVM) has a lot to offer for the treatment of feline colitis. This ancient holistic approach aims to restore balance and harmony within the body by addressing the underlying patterns of disharmony or imbalance. The primary modalities used in TCVM (see sidebar on page 50) are acupuncture, herbal medicine, food therapy, and Tui-na (Chinese massage). This article looks at how these modalities can help feline patients with colitis.

PATTERNS OF DISHARMONY IN FELINE COLITIS In cases of feline colitis, TCVM practitioners assess the animal based on four diagnostic methods: inspection, listening and smelling, inquiry, and palpation. They then identify patterns of disharmony associated with the condition, which often include: 1. Spleen Qi Deficiency 2. Liver Qi Stagnation 3. Damp-Heat in the Large Intestine 4. Kidney Yang Deficiency 5. Blood Stasis 6. Cold-Damp Invasion

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While these disharmonies can affect any cat, they are more common in those of certain ages, breeds, and predispositions.

1. S PLEEN QI DEFICIENCY A deficiency in the energy required to transform and transport nutrients results in digestive issues. Weak digestion, loose stools or diarrhea, and a lack of energy characterize this pattern. Cats may also exhibit poor appetite, weight loss, and a pale tongue with a thin white coating.

Susceptibilities • A ge-related changes in the digestive system can contribute to decreased nutrient absorption and weakened Spleen Qi. • Some breeds are predisposed to, or have a history of, gastrointestinal issues, obesity, chronic disorders, or metabolic disorders.

• A sedentary lifestyle or obesity weaken the Spleen's function.

• Overfeeding or providing a diet high in processed or difficultto-digest plant protein and/or high-carbohydrate foods can contribute to the problem.

Treatment Supporting digestion and strengthening the Spleen is essential.

• A cupuncture points: ST36 (Zusanli), SP6 (Sanyinjiao), CV12 (Zhongwan), BL20 (Pishu) • H erbal formulas: Shen Ling Bai Zhu San, Bu Zhong Yi Qi Tang • F ood: should be warm and easy to digest, including lean proteins like chicken or turkey and cooked vegetables like pumpkin or sweet potato. Avoid raw vegetables or cold foods, as they can exacerbate the deficiency.

2. LIVER QI STAGNATION This pattern is often seen in cats experiencing stress or emotional disturbances. Stressful environments, lack of mental stimulation, or confinement in small spaces can increase stress and anxiety, leading to Qi stagnation. Symptoms include irregular bowel movements, alternating diarrhea and constipation, and abdominal pain. Cats may also show signs of irritability, depression or anxiety.

Susceptibilities • M iddle-aged or older cats with prolonged stress, emotional disturbances, or an unhealthy lifestyle can be susceptible. As cats age, their ability to handle stress may decrease, leading to stagnant liver energy.

• S ome breeds are predisposed to stress, anxiety, or behavioral issues.

• C ats with a history of liver disease, hormonal imbalances, or chronic stress are prone to this pattern.

• C ats with breed-specific liver or metabolic issues could also be at a higher risk.

Treatment The focus is to soothe and relax the Liver while promoting the smooth flow of Qi. • A cupuncture points: LR3 (Taichong), GB34 (Yanglingquan), ST25 (Tianshu), BL18 (Ganshu) • H erbal formulas: Xiao Yao San, Chai Hu Shu Gan San • T ui Na • F ood: provide those with calming effects, like leafy greens and lean proteins such as turkey. Avoid feeding overly rich or greasy foods, as they can contribute to stagnation.

3. DAMP-HEAT IN THE LARGE INTESTINE This pattern is characterized by inflammation and infection in the colon, leading to symptoms like diarrhea with foul-smelling, mucus-laden stools, increased thirst, fever, and a red tongue with a yellow, greasy coating. Damp-Heat accumulates in the colon, leading to inflammation.

Susceptibilities • Y ounger cats with weaker immune systems may be more susceptible to infections that can develop Damp-Heat.

• O lder cats with weakened immune systems may also be at risk due to age-related changes.

• B reeds predisposed to infections, immune system issues, or skin allergies may be more susceptible. Certain breeds are predisposed to inflammatory bowel disease, which can be associated with Damp-Heat in the Large Intestine.

• P oor hygiene, exposure to environmental toxins, and lack of access to clean water can increase a cat’s susceptibility to infections and inflammation, with potential for Damp-Heat.

• A history of infections, inflammatory bowel disease, immune system issues, and prior medical conditions that have compromised the cat’s immune system or gastrointestinal health also raise the risk.

Treatment The focus here is on clearing Heat and Dampness.

• A cupuncture points: ST25 (Tianshu), LI11 (Quchi), LI4 (Hegu), ST37 (Shangjuxu), CV6 (Qihai) • H erbal formulas: Ge Gen Huang Lian Huang Qin Tang, Ba Zheng San • F oods: Cooling foods like rabbit and whitefish are best.

4. K IDNEY YANG DEFICIENCY This pattern leads to an inability to warm and transform food, resulting in digestive dysfunction. Symptoms include loose stools, diarrhea with undigested food particles, cold extremities, and lethargy. A weak lower back and pale tongue are also characteristic of this pattern.

Susceptibilities • K idney energy naturally declines with age and can lead to weakened Kidney Yang, making older cats more susceptible to chronic conditions and imbalances related to this pattern.

• C ertain breeds may have a higher risk of developing kidney IVC Summer 2023

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THE FOUR PRIMARY MODALITIES OF TCVM Acupuncture Acupuncture involves the insertion of thin needles at specific points on the body to stimulate the flow of Qi and blood, promote healing, and restore balance. Acupuncture points are chosen based on identified patterns. This modality can help alleviate the inflammation, pain, and gastrointestinal symptoms associated with feline colitis, and is tailored to each cat's unique pattern and symptoms.

issues that contribute to this pattern. For example, Persian cats are prone to polycystic kidney disease.

• A cold or damp living environment impairs the body’s ability to generate and maintain warmth.

• A diet low in protein or essential nutrients can weaken Kidney Yang.

• C ats with a history of kidney disease, urinary tract issues, or hormonal imbalances are at higher risk.

• P rior medical issues impacting kidney function or overall health also raise susceptibility.

Treatment Supporting Kidney function is crucial for this pattern.

• A cupuncture points: BL23 (Shenshu), KI3 (Taixi), GV4 (Mingmen), CV4 (Guanyuan) • H erbal formulas: Jin Gui Shen Qi Wan, You Gui Wan • F ood: give warming foods like cooked lamb, beef, chicken, and small amounts of ginger. Avoid cold and raw foods, as they can further weaken Kidney Yang.

Herbal medicine Herbal formulas are also tailored to the cat’s specific patterns, and can help strengthen the immune system, reduce inflammation, and address underlying imbalances. The herbs are carefully selected and combined to provide a synergistic effect and address the root cause of the colitis.

Food therapy In food therapy, the cat’s diet is adjusted to support overall health and address the identified patterns. Foods with specific energetic properties are recommended based on the cat's condition, with the goal of strengthening the immune system, improving digestion, and alleviating the symptoms related to colitis. Specific ingredients and cooking methods are recommended, again based on the cat's pattern and needs.

Tui-na A form of Chinese massage, Tui-na stimulates specific acupressure points and meridians in the cat's body, especially when needle insertion is difficult. It can be used alongside acupuncture and herbal medicine to enhance their effects and support the cat. Tui-na is also an effective followup therapy for cats between appointments.

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5. B LOOD STASIS This pattern can result from long-term inflammation or injury to the colon, as blood circulation is impaired. Symptoms include constipation or diarrhea with dark, tarry stools, abdominal pain, a purple tongue with dark spots, and severe abdominal pain.

Susceptibilities • O lder cats are prone due to the increased likelihood of longterm inflammation, injury, or chronic conditions. Age-related changes in circulation and blood vessel elasticity can also contribute.

• C ertain breeds are predisposed to circulatory issues, cardiovascular disorders, or clotting problems.

• C ats with a sedentary lifestyle, or those recovering from injury or surgery, may have reduced circulation.

• C hronic stress or exposure to environmental toxins can contribute to inflammation and blood stasis.

• A history of circulatory issues, cardiovascular conditions, clotting disorders, or long-term inflammation can also be a contributing factor.

• P revious medical conditions may impact the cat’s blood circulation or vascular health, and lead to the development of this pattern.

Treatment Promoting blood circulation and removing blood stagnation is essential.

• A cupuncture points: SP10 (Xuehai), BL17 (Geshu), LI11 (Quchi), ST37 (Shangjuxu), SP6 (Sanyinjiao) • H erbal formulas: Tao Hong Si Wu Tang, Xue Fu Zhu Yu Tang • F oods: focus on lean proteins like chicken or turkey, dark leafy greens, and small amounts of turmeric.

6. C OLD-DAMP INVASION This pattern is characterized by the sudden onset of diarrhea, which may be watery or contain mucus, along with abdominal pain. Cats may also exhibit a lack of appetite, vomiting, and a pale tongue with a white, greasy coating.

Susceptibilities • Y ounger or older cats with weakened immune systems may be more vulnerable to external pathogens that can lead to this pattern. Age-related changes in the digestive system make older cats more prone.

• B reeds predisposed to gastrointestinal issues, weak immune systems, or skin allergies, as well as those with specific dietary sensitivities, are susceptible.

• L iving in a cold, damp environment, or eating a diet high in raw or cold foods, are risk factors.

Treatment The goal here is to expel cold and dampness from the body.

• A cupuncture points: ST25 (Tianshu), ST36 (Zusanli), CV6 (Qihai), SP9 v(Yinlingquan), BL20 (Pishu) • H erbal formulas: Li Zhong San, Wu Mei San • F ood: give warming and drying foods like cooked chicken, turkey, and pumpkin.

From the VMAA The Veterinary Medical Aromatherapy® Association (VMAA) is an organization of veterinarians and veterinary technicians dedicated to the responsible use of aromatherapy in animal practices. Their mission is to promote standards of excellence in animal aromatherapy, to provide outreach and education to veterinarians and auxiliary animal practices, and to promote continual improvements in Veterinary Medical Aromatherapy®. The VMAA is elevating the veterinary profession through innovation, education, and advocacy of integrative medicine.

GI-FRIENDLY ESSENTIAL OILS Digestive problems such as IBS-D, gastroenteritis, colitis, constipation, and colic are often seen in both large and small animals. Essential oils produced from various plants and herbs have applications in veterinary medicine, and can be used to help with these conditions.

The following essential oils may help treat various GI conditions encountered in practice:

1. P eppermint (Mentha piperita): The spasmolytic effect of this oil has been well documented, making it a good choice for topical and/or oral administration.1-3 Applying a small amount at BL 25, ST 36, or Guan Yuan Shu prior to needle insertion, or with acupressure, can be helpful for mitigating symptoms.

2. Fennel (Foeniculum vulgare): Known for its carminative action, even the inhalation4 of fennel oil can help relieve discomfort from gas in the intestinal tract, as well as flatulence. This oil has been shown to have action against H. pylori, so consideration for oral use may be appropriate. 5 Fennel is often combined with other oils such as ginger, peppermint, tarragon, or lemongrass. Since fennel is considered warming, dilution is appropriate for topical use.

3. P atchouli (Pogostemon cablin): Here is an essential oil not often considered for GI issues. The anti-inflammatory action of patchouli is significant and may help diminish the effects of inflammation in IBS-D cases. 6 In rats, patchouli oil combined with tangerine oil (1:2 ratio), has been observed to have a protective effect against gastric ulcers. 7 Sedation from inhalation may help with overall comfort. 8

When using TCVM to treat a cat with colitis, it’s crucial to identify and understand these specific patterns of disharmony.

A TCVM practitioner can then create a comprehensive plan utilizing several or all modalities at their disposal to restore balance and harmony within the cat’s body.

Submitted by Susan Albright, DVM

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From the VBMA

The Veterinary Botanical Medicine Association is a group of veterinarians and herbalists dedicated to developing responsible herbal practice by encouraging research and education, strengthening industry relations, keeping herbal tradition alive as a valid information source, and increasing professional acceptance of herbal medicine for animals.

DID YOU GUESS LAST ISSUE’S MEDICINAL HERB? The flowering tops of feverfew (Tanacetum parthenium) are indicated for clearing Liver Fire, thanks to the herb’s strong anti-inflammatory influence. Feverfew is popular for preventing and treating migraine headaches or Liver Fire head pain. It is a mild nervine, useful for nervous headaches and stress-induced GI symptoms, and exhibits anxiolytic and antidepressant-like effects.

CASE REPORT Geppetto is a seven-year-old male castrated DSH cat. When he was two, he suffered from a urinary blockage complicated by severe varicosities in the urethral area.

Once the varicosities were shrunk with the use of yarrow (Achillea millefolium), horse chestnut (Aesculus hippocastanum) and lady’s mantle (Alchemilla vulgaris), he regained his health. This past year, however, Geppetto started to show symptoms of headaches, with persistent episodes of significant head pressing. Normal blood work and physical exam ruled out metabolic issues. Given his past vascular issues, a migraine-like headache was suspected. Within two weeks of receiving feverfew, the severity of Geppetto’s head pressing was reduced by over 90%.

PAST WEBINARS • VBMA’s first webinar of 2023 featured Dr. Barbara Fougere with “A Practitioner’s Panel on Feline Respiratory Disease”, including asthma and chronic upper respiratory disease. Among the favorite herbs she has used in the treatment of asthma are Marshmallow (Althea officinalis), Perilla seed (Perilla frutescens), and licorice (Glycyrrhiza glabra). When discussing chronic URD, Dr. Fougere mentioned additional herbs such as Solidago (Solidago spp.), Astragalus (Astragalus membranaceous) and mushroom complexes.

Dr. Fougere stressed that environmental issues and stress must also be addressed; glycetracts can be used to increase patient

compliance; marshmallow, mushrooms, and Lyprinol are very valuable; and a healthy gut microbiome is an important factor in achieving health.

• Dr. Rona Sherebrin addressed the fear of using essential oils in her webinar. Cats do have issues with phenols due to slower Phase 2 hepatic conjugation, so she advised against the use of essential oils like wintergreen, bitter almond, garlic, horseradish, pennyroyal, and the citrus family. Dr. Sherebrin encouraged the use of diffusers, with short time-controlled sessions. She also shared a formula of balsam fir (Abies balsamea), copaiba (Copaifera officinalis), lemon balm (Melissa officinalis), and Frankincense (Boswellia serrata).

Using pattern differentiation, Dr. Sherebrin discussed her favorite formulas: San Ren Tang, Three Seed Decoction, which is a bland formula and well tolerated; Xin Yi San, Magnolia Flower formula; and Yu Ping Feng when there are extra signs of wind.

• Dr. Cynthia Lankenau discussed a chronic stuck feline herpes case that responded beautifully to a Gu formula. Gu is an ancient Chinese syndrome that is synonymous with chronic conditions caused by endotoxins and microbes hiding in biofilm.

OTHER EVENTS From June 9 to 11, the International Herbal Symposium took place at Wheaton College, and was also live-streamed. The American College of Veterinary Botanical Medicine (ACVBM) held an onsite pre-IHS conference. A review of these events will be provided in next issue’s column.

Submitted by Cynthia Lankenau, DVM

GUESS THIS HERB!

Join the VBMA at vbma.org to find out. The answer will also be published in the next issue of IVC Journal.

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Factors influencing the equine

microbiome

— and how to support it

BY SUUS BETTINK, DVM

“No feet, no horse.” This old saying says it all, and can be extended to gut health as well. In fact, I would dare say: “No gut, no horse.” If a horse doesn’t have optimal gut microbiome health, they won’t have optimal overall health.

The equine world is increasingly recognizing this truth, but we still have a long way to go, and the way we care for our horses in general can interfere with achieving good gut microbiome health. It’s vital to have an understanding of the equine microbiome, along with what causes it to get out of balance and how you can support it to reestablish well-being.

HORSES ARE HINDGUT FERMENTERS The horse is capable of utilizing a wide range of plant species1 as a hindgut fermenter. This fermentation is done by a complex microbial system in the gut. The bacteria, viruses, archaea, and fungi that make up the microbiome thrive on high fiber, low energy forage.2 The microbiome helps transform non-digestible fiber into short chain fatty acids to be used by the immune system to lower inflammation; as an energy source for the intestinal cells; and to repair the gut lining.

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MICROBIOME DIVERSITY IS CRUCIAL When the diversity of organisms in the microbiome decreases, chronic illnesses appear. A variety of factors can change the microbiome, and these alterations are very complex to understand.

All the research that has been done involves equine feces, and it’s unclear if they completely represent the microbiome in the hindgut. In addition, the scope of human microbiome research is shifted to the metabolome — the study of metabolites, which could include lipids, amino acids, short peptides, sugars, alcohols or organic acids. This means, for example, not examining the bacteria in the gut, but what they are transforming and how it will benefit or harm the host.

What the research makes clear is that the equine microbiome is dramatically changed if the horse has diseases such as laminitis or severe colic. But there is another way to look at it. I think the lack of diversity in the equine microbiome, whatever the reason behind it, is causing chronic illnesses like metabolic diseases, PPID, and others to develop.

High stress levels and a lack of forage diversity are among the factors causing imbalances in the equine microbiome.

A horse is meant to walk several miles a day and search for diverse foods, including different grasses and herbs and the tough little branches on trees. This diversity in forage leads to diversity in the gut microbiome. The more diverse the gut bacteria, the less disease will occur.

In humans, we can see how unhealthy lifestyles are impacting overall health. With less diversity in our microbiomes, more diseases are arising.3

PROTON PUMP INHIBITORS HARM GUT HEALTH

THE GUT-BRAIN AXIS AND STRESS More and more human studies are being done on the gutbrain axis, showing that the vagus nerve plays an important role. The gut interacts with the brain cells, and vice versa.

Chronic stress undermines the gut microbiome. The HP axis is continually activated and the sympathetic nervous system is in overdrive, resulting in increased cortisol levels. This results in immunodeficiency and chronic intestinal illnesses.

The number of gastric ulcers our horses have can also be a sign we humans are asking too much of them. If a horse is on Gastroguard, try to wean them off it as quickly as possible because proton pump inhibitors are very harmful to gut health. They lower the stomach pH, which can cause pathogenic bacteria to enter the duodenum and jejunum and cause problems there. Also, if the gut is not healthy, the uptake of vitamin B12 is lowered; this can cause health problems because food isn’t digested properly, leading to an overgrowth of bad bacteria in the jejunum.

Studies in mice and human athletes have shown that endurance tests and involuntary exercise harm the gut microbiome, while

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voluntary exercise benefits it.7,8 A study done in horses, meanwhile, showed that demanding endurance tests caused increased intestinal permeability; raised pro-inflammatory interleukines; and produced LPS (lipopolysaccharide), an endotoxin that can enter the bloodstream and cause more harm to the immune system and microbiome.9

Guard against obesity. A lot of horses are too fat.

4. They eat too many sugars and don’t get enough pre-

biotics, the insoluble starches. Obesity indicates an imbalance in the gut microbiome. Every craving you have is actually the microbes in your gut speaking to your brain (gut-brain axis); they want a certain type of food and are signaling to get it in order to grow and to stay alive.

HOW TO SUPPORT THE EQUINE MICROBIOME

In humans with obesity, the ratio between bacteroidetes and firmicutes has changed. The proportion of bacteroidetes is reduced11 — bacteroidetes transform non-digestible carbohydrates such as pectine, oligosaccharide and insulin. This research has also been done in horses with the same results seen in the ratio between bacteroidetes and firmicutes.11 In humans, supplementing with Akkermansia muciphinilia shows improvement in obesity and metabolic disease.12

1. reducing stress levels. If a horse doesn’t want to work,

Look at how the horse is managed, with the goal of try to figure out why not by listening more carefully to them. Don’t look on the horse as a possession. Horse and human comprise a partnership, in which they co-exist with one another. Look at the horse’s social interactions with both friends and bullies within the herd. Can the horse escape from a bully, and find comfort from companion horses? Is the work you do with the horse good for them? Is it too monotonous, too difficult, or too restraining? Is the horse noncompliant or submissive with bit, spurs and side reins? In summary, take a closer look at the horse and the behavior they’re showing.

In the study done by Hu et all, firmicutes were found to be higher in captive Przewalski horses than in wild ones.6 Also, Provenza’s research showed that if an animal lacks a certain mineral or vitamin, they will overeat to compensate for the deficiency. It’s better to give whole foods instead of supplements to help with a deficiency. So keep this in mind if you want to lower a horse’s weight.

Try to get the horse in balance by enhancing the gut-

2. brain axis and giving attention to the vagus nerve.

This balance can be achieved by osteopathy and/or acupuncture. The communication between the brain and the gut will increase, cortisol levels will drop, and the horse’s health will reestablish.

5.

Make sure the horse eats slowly and without stress. If a horse eats slowly enough, saliva is produced and the digestion process starts. Eating slowly also ensures the food is broken down into very small particles, which increases digestion in the stomach. Stress causes digestion to decrease so the peptides can’t be properly broken down into amino acids. This can cause SIBO (small intestinal bacteria overgrowth) because the proteins can be used by the bad bacteria in the jejunum, causing the overgrowth. Also, the food bolus stays in the stomach longer and causes problems in the duodenum.

6.

Ensure the horse drinks enough water. This helps the digestive system and detoxes the body. It’s important the horse has access to fresh, ample drinking water.

Look at the horse’s forage. As far as I’m concerned,

3. few horses need concentrate feed. Good hay or grass

and quality mineral sources are enough. In his 40 years of study, Provenza found that animals need good plant diversity to achieve optimal biochemical composition in the body and obtain good health. He stated that animals eat 50 to 75 different plant species in one meal. So don’t feed horses a monoculture diet. The quality of the food is also important, so no hay or grass that has been sprayed with herbicides like glyphosate, or is full of heavy metals due to pollution.

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7. might affect the gut microbiome. Try to deworm with

Think about the medications you use and how they natural products and good pasture management. If that’s not doable, try to deworm as little as possible after an egg count.

If a horse’s stress level is reduced, and their forage optimized, yet there is still need for intervention, decreasing inflammation, helping the body detox, and eliminating pathogens will also help build a good gut microbiome.

1Costa MC, Weese JS. The equine intestinal microbiome. Anim Health Res Rev; Conference

of Research Workers in Animal Diseases; 2012. pp. 121–128.

2Dougal K, Harris PA, Edwards A, Pachebat JA, Blackmore TM, Worgan HJ, Newbold CJ.

A comparison of the microbiome and the metabolome of different regions of the equine hindgut. FEMS Microbiol Ecol. 2012;82:642–652.

3Blaser Martin J., Falkow Stanley. What are the consequences of the disappearing

human microbiota? Nat Rev Microbiol. 2009 Dec; 7(12): 887–894.

4Li Y., Zhang K., Liu Y., Li K., Hu D., Wronski T. Community composition and diversity

of intestinal microbiota in captive and reintroduced Przewalski’s horse (Equus ferus przewalskii) Front. Microbiol. 2019;10:1821. doi: 10.3389/fmicb.2019.01821.

5Provenza, Fred. Nourishment: what animals can teach us about rediscovering our

nutritional wisdom.

6Dini Hu, Chen Wang, Make Ente, Ke Zhang, Dong Zhang, Xuefeng Li, Kai Li, and

Hongjun Chu, Assessment of Adaptation Status of Reintroduced Equus Przewalski Based on Comparative Analysis of Fecal Bacteria with Those of Captive E. Przewalskii, Domestic Horse and Mongolian Wild Ass, Animals (Basel). 2022 Oct; 12(20): 2874.

7 Clark Allison, Mach Núria, Exercise-induced stress behavior, gut-microbiota-

brain axis and diet: a systematic review for athletes, J Int Soc Sports Nutr 2016 Nov 24;13:43.

8 Megumi Matsumoto, Ryo Inoue, Takamitsu Tsukahara, Kazunari Ushida,

Hideyuki Chiji, Noritaka Matsubara, Hiroshi Hara, Voluntary running exercise alters microbiota composition and increases n-butyrate concentration in the rat cecum Biosci Biotechnol Biochem 2008 Feb;72(2):572-6.

9 Núria Mach, Yuliaxis Ramayo-Caldas, Allison Clark, Marco Moroldo,

Céline Robert, Eric Barrey, Jesús Maria López and Laurence Le Moyec, Understanding the response to endurance exercise using a systems biology approach: combining blood metabolomics, transcriptomics and miRNomics in horses, BMC Genomics. 2017; 18: 187.

10 Ley R. E., Turnbaugh P.J.,Klein S. and Gordon J.I. (2006), Human gut

microbes associated with obesity, Nature 444, 1022-1023.

11Amy S. Biddle,* Jean-Francois Tomb and

Zirui Fan, Microbiome and Blood Analyte Differences Point to Community and Metabolic Signatures in Lean and Obese Horses, Front Vet Sci. 2018; 5: 225.

12Vanessa

Fernandes Rodrigues, Jefferson Elias-Oliveira, Ítalo Sousa Pereira, Jéssica Assis Pereira, Sara Cândida Barbosa, Melissa Santana Gonsalez Machado, and Daniela Carlos, Akkermansia muciniphila and Gut Immune System: A Good Friendship That Attenuates Inflammatory Bowel Disease, Obesity, and Diabetes, Front Immunol 2022 Jul 7;13:934695.

WILD VERSUS DOMESTIC HORSES According to research by Li et al,4 Przewalski horses have a more diverse gut microbiome. The researchers hypothesize it’s because they have access to different and more diverse forage. I endorse this, but I also think Przewalski horses have lower stress levels, which could also play a role in their microbiome diversity.

Overall, horses in our Western world need to act when we humans want them to. They spend time in stables, and need to be ready to perform when our schedules tell us.

However, there are some questions we should be asking.

Are our horses allowed to be themselves in competition as well as in recreation? Can they rest? Can they socially interact with other horses? Do they have the freedom to walk away or to say “no” to jobs? Can they eat when they want, and can they choose the type of forage they consume?

Research done by Fred Provenza5 shows that animals choose their food, and also self-medicate. In addition, no two animals select the same combination of ingredients, nor the same food, from day to day. A horse can eat like this in the wild, but not if they’re in a stable or pasture, unless we provide the necessary food diversity. And according to Provenza, when we break our links with our ancestors, they’re very difficult to reestablish. This means it’s hard to get back to health if you’ve moved far from your original food and microbiome. Ideally, you should eat like your ancestors did. It’s hard to establish a horse’s true ancestor, however, since a big genetic melting pot has been created in breeding horses. Studies done with Przewalski horses also show that genes adapt more slowly than phenotypes, and these horses need human intervention when reintroduced to the wild.4

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Updated information and assistance from experts help veterinary teams successfully treat both common and uncommon endocrine problems.

New endocrine guidelines from Endocrine disorders are among the most prevalent chronic diseases seen in veterinary practice, and identifying and managing them can be difficult and daunting. To help enhance diagnosis and treatment, the 2023 AAHA Selected Endocrinopathies of Dogs and Cats Guidelines were recently released by the American Animal Hospital Association (AAHA). This free resource offers easy-to-use reference tools and straightforward procedures to identify and treat endocrine problems in veterinary practice.

The guidelines were created by a group of veterinary professionals, and are based on the most recent academic findings and clinical practice. They provide useful and comprehensive strategies to assist veterinary teams

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AAHA

in dealing with these challenging conditions. Since endocrinopathies can pose special obstacles to the timeconstrained practitioner, the guidelines list diagnoses and treatments using simple, step-by-step procedures. They offer current clinical suggestions to help teams stay informed and deliver the best care possible for a variety of endocrine problems.

The most prevalent endocrine diseases in dogs and cats are included in this resource. It’s also helpful for less frequently-seen feline conditions, including hypothyroidism and hyperaldosteronism. This streamlined approach for the veterinary team can help cut any possible confusion and support practitioners in doing their very best for their endocrine patients.

The American Animal Hospital Association (AAHA) has been accrediting veterinary hospitals across the US and Canada since 1933, using criteria directly tied to providing high quality medical care and humane treatment.

Visit www.aaha.org/endocrine-disease to download the guidelines.

Veterinary professionals are vulnerable to stress — 7 WAYS TO EASE THE ANGST

BY OMER RASHID, DVM

Stress is a serious problem among veterinarians and their staff, and can lead to anxiety, depression, and other issues if not addressed.

Recognizing the signs and using one or more of the calming techniques highlighted in this article will help you gain control of your stress before it becomes overwhelming.

As a busy veterinary practitioner, you’re probably no stranger to stress, whether it arises from working long hours or dealing with upsetting situations. Knowing how to recognize stress — and take steps to alleviate it — is vital to your mental and emotional well-being.

RECOGNIZING THE SYMPTOMS

• Non-existent sense of humor • Inability to enjoy activities that would normally make you happy • Feelings of isolation Stress can also manifest physically in the following ways:

Once the body is in the stress response phase, hormones such as adrenaline and cortisol are released and the muscles tighten.

Tension headaches, back pain, anxiety, indecisiveness, poor focus, and irritability are some of the symptoms that can ensue. It’s not surprising, then, that a stressed state of mind not only reduces your efficiency, but also hampers your ability to deal with challenges.

We all experience stress in various ways. It’s useful to be able to recognize the early indicators before things spiral out of hand.

Common emotional stress indicators include: • • • •

ension T Agitation or hostility Inability to unwind General lack of interest in life

• • • • •

Hyperventilating or shallow breathing Tense muscles Trouble sleeping Headaches Digestive problems such as diarrhea, vomiting, and constipation • Sexual issues • Grinding teeth or clenched jaw • Weariness or lack of vigor Let’s look at seven techniques — validated by psychologists — to combat stress. Using one or more of these techniques will not only help you feel better, but also increase your ability to meet the mental and emotional demands of veterinary practice.

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1. TRY MEDITATION

4. PRACTICE STOICISM

Meditation includes various techniques conducive to mindfulness, such as deep breathing. These techniques can prove immensely helpful in countering stress. If done correctly, they can help you stay calm so you can go about your work routine without feeling frazzled.

This is a great way to stay calm. The idea is to train yourself to be unperturbed even in the face of extremely stressful situations.

The philosophy of stoicism is based on the principle that a person should know what is possible, while being aware of what is insignificant. Stoicism helps you believe that while you may not have control over the events affecting you, you can control how you handle them. Practising stoicism can help you achieve serenity and a stable personality, and assist you in responding better to the stressful situations that may come your way.

• B reath focus: The idea is to put all your concentration on your breathing. For this powerful stress-relieving technique, take long, deep, slow breaths. Also known as abdominal breathing, this focused way of breathing disengages you from stressful thoughts, and makes you feel fresh and calm.

• B ody scan: In this technique, deep breathing is combined with focus on a particular body part. For instance, if you feel stress around your neck area, focus on it while practicing abdominal breathing, and mentally release the tension. This will give you immediate relief from the feeling of tight muscles caused by stress.

• G uided imagery: Using your mind to conjure up pleasant images of soothing experiences can be a relaxing and rejuvenating meditation exercise. You can find online apps with a repository of calming images to be recalled while meditating.

2. HAVE A QUICK NAP Enjoying a power nap at a designated spot in the workplace will diminish stress. Identify an area that is likely to be always available for your nap. You can also arrange to play soothing music in that spot to help you doze off. Studies have shown that a short nap can improve work performance for almost four hours. Even a tenminute nap can do wonders when it comes to relieving stress and starting afresh.

3. TAKE A BREAK If things become too much to handle, and the stress seems to be getting the better of you, consider taking a break. We often tend to associate stress with the environment we are working in, so taking a break from it can be helpful. Go to the beach, or another peaceful place, or just relax at home. Don’t be afraid to talk to the person in charge about how you are feeling and why you need a break. It is only human to experience a stress response that requires you to have a temporary breather from work.

The prevalence of burnout and stress in veterinarians Recent data suggests that burnout and stress are climbing in veterinarians. Results collected from the AVMA Census of Veterinarians from 2016 to 2020 show burnout has consistently risen year over year, almost doubling from 11.8 % in 2016 to 22% in 2020.1 A 2021 study conducted jointly by Merck Animal Health and the American Veterinary Medical Association (AVMA) revealed that the percentage of veterinarians with serious psychological distress (as measured by the Kessler Psychological Distress Scale) hit 9.7% in 2021, as compared to 6.4% in 2019. The prevalence of severe psychological distress among support staff was nearly twice as high (18.1%) compared to two years ago. Additionally, half of staff respondents (49.6%) and approximately one-third (30.5%) of veterinarians reported high levels of burnout.2

1Neill Clinton, Hansen Charlotte, Salois Matthew (2022). The Economic Cost of

Burnout in Veterinary Medicine, JOURNAL Frontiers in Veterinary Science Vol 9 2022, www.frontiersin.org/articles/10.3389/fvets.2022.814104, DOI=10.3389/fvets.

2022.814104.

2Burnout, stress plague veterinary industry amidst pandemic. Veterinary Practice

News. www.veterinarypracticenews.com/burnout-stress-plague-veterinary-industryamidst-pandemic/. Published February 10, 2022. Accessed March 23, 2023.

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5. CONSIDER TAI CHI, QIGONG, OR YOGA Learning any of these ancient arts can help you remain calm and stress-free. They combine rhythmic breathing with flowing postures, and have benefited many people. Practicing these arts also distracts you from stressful thoughts and supports mental focus.

6. SET GOALS Recognizing stress and being able to remain calm is one of the most valuable skills a person can possess. Something to help you achieve this quality is goalsetting. By firmly setting goals and sticking to them, you give yourself a positive sense of commitment, leading to an optimistic approach to all aspects of life. This optimism results in an overall happier and stress-free demeanor. You can set health, professional, or business goals to keep you motivated. It will help you be more productive as well as positive, leading to a feeling of satisfaction and fulfillment, and making you less prone to stress.

7. BE POSITIVE It’s crucial to try and develop a positive outlook when handling veterinary stress. Using alcohol or drugs to reduce tension is a vicious cycle. In fact, it may prolong the body’s stressed condition rather than calming it down, and will ultimately result in more problems.

Maintaining a positive outlook can motivate you to deal with stress in a better way. This approach also encourages you to ask for help if you need it. It’s critical to keep in mind that consulting a doctor or mental health professional for assistance is nothing to be ashamed of. Finding more constructive ways to deal with stress is all that is necessary to prevent it from having a negative impact on your health, or the health of those around you.

Stress can easily spiral out of control and lead to other problems such as anxiety and depression. By recognizing the signs of stress, and adopting one or more of the calming techniques outlined in this article, you can get a handle on your stress before it gets a handle on you.

How positive and negative coping mechanisms affect performance It’s important to keep in mind that emotional stress can sometimes be empowering and facilitate better performance, since concentration, focus and efficiency are improved. The typical "fight or flight" reaction to perceived threats is where this phenomenon is most obvious, although there is a breaking point at which too much stress compromises performance.

People who appreciate challenges, achieve states of heightened awareness, and thrive in stressful circumstances are drawn to the area of surgery. However, a constant use of healthy stress management techniques is necessary for long-term success in this area.

Although there is a wealth of general research on the relationship between stress and performance, less is known about the specific impact of stress on surgical practice. One qualitative study1 examined key surgical stresses, their effects on performance, and the coping mechanisms employed by surgeons. The goal was to investigate the impact of surgical stress and the essential components of efficient coping mechanisms.

Sixteen interviews were conducted for the study, with consultants and trainee surgeons among the participants. Junior surgeons expressed concern about their ability to handle stressful surgical situations, whereas older surgeons had well-rounded techniques in place for handling surgical stress. These results revealed that despite the serious consequences stress poses, coping mechanisms are not expressly taught to surgical trainees, which means they are not well prepared and therefore more prone to making errors.

Effective positive coping techniques for stressful situations include minimization, distraction, situation management, substitute gratification, reaction control, and positive self-instructions. Having the ability to employ positive coping mechanisms like these helps us perform better under pressure.

1Wetzel CM, Kneebone RL, Woloshynowych M, et al. The effects of stress on surgical

performance. Am J Surg. 2006;191(1):5-10. doi:10.1016/j.amjsurg.2005.08.034.

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HOW UPDATED STATE LAWS COULD CHANGE VIRTUAL VETERINARY CARE Legalizing the virtual veterinarian-clientpatient relationship (VCPR) will enhance veterinary care in Arizona and beyond.

Virtual veterinary care has become more common since the pandemic. However, this approach can make it challenging to negotiate the standards and laws around telemedicineʼs veterinarian-client-patient relationship (VCPR). According to the American Veterinary Medical Association (AVMA), a VCPR is present when all the following conditions are satisfied:

1.

2.

he client has agreed to T abide by the veterinarian’s instructions and has given the veterinarian permission to make any clinical decisions regarding the patientʼs health.

If the veterinarian is familiar enough with the patient, he or she can make a basic or preliminary diagnosis of their medical state. By examining the patient, or seeing and handling the animal during timely and medically appropriate visits to the facility, the veterinarian has demonstrated that they are personally familiar with the patient's care.

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3.

4.

5.

he veterinarian has made T arrangements for veterinary emergency coverage, ongoing care, and treatment, or is easily available for follow-up evaluation.

The veterinarian oversees the course of treatment, adherence, and result.

Records are kept on the patient.

On May 15, Arizona's Senate Bill 1053 was signed into law by Governor Katie Hobbs. The bill permits Arizonalicensed veterinarians to set up a VCPR using telemedicine, and also lets them write short-term, non-controlled substance prescriptions.

According to the statistics, the number of animals surrendered to Arizona shelters by their owners for medical reasons rose by 13% in just the last year. Meanwhile, the first

appointment a pet owner can get at a veterinary hospital is usually six weeks away. This new law will lessen unnecessary suffering for pets and help maintain them in caring homes.

Arizona is one of just a few states to put this life-saving legislation into effect.

The American Veterinary Medical Association drafted the final amended version of the law, which was approved by a nearly unanimous vote and lays important groundwork for other states. www.dvm360.com/view/updateson-state-legislation-for-veterinaryvirtual-care