Dairy Planner April -Issue 2019 by PIXIE CONSULTING SOLUTIONS LTD.

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HARBIL/2004/22481

Vol.16 | No. 4 | April - 2019

From the Pen of Chief Editor

Editorial

Role of technological processes and potential impacts on dairy The microstructure of milk fat in processed dairy products is poorly known despite its importance in their functional, sensorial and nutritional properties. However, for the last 10 years, several research groups including our laboratory have significantly contributed to increasing knowledge on the organization of lipids in situ in dairy products. This paper provides an overview of recent advances on the organization of lipids in the milk fat globule membrane using microscopy techniques (mainly confocal microscopy and atomic force microscopy). Also, this overview brings structural information about the organization of lipids in situ in commercialized milks, infant milk formulas and various dairy products (cream, butter, buttermilk, butter serum and cheeses). The main mechanical treatment used in the dairy industry, homogenization, decreases the size of milk fat globules, changes the architecture (composition and organization) of the fat/water interface and affects the interactions between lipid droplets and the protein network (concept of inert vs active fillers). The potential impacts of the organization of lipids and of the alteration of the milk fat globule membrane are discussed, and technological strategies are proposed, in priority to design biomimetic lipid droplets in infant milk formulas.

C O N T E N T S A way of scientiďŹ c management of foot and mouth disease (fmd) in ...

Leptospirosis - an overview

Etiopathogenesis and different approaches for bovine ketosis

Mastitis in dairy : prevention can save costly treatment

SNP chip for indigenous cattle and buffalo

Tea waste as an animal feedstuff

Recipe

News

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EDITORIAL BOARD MEMBER Dr. J Tamizhkumaran M.VSc., PGDEP., Ph.D. (Ph. D in Veterinary & Animal Husbandry Extension Education)

Dr. Anjali Aggarwal Principal Scientist Dr. Sanjay K Latkar Alembic Pharmaceuticals Ltd Mumbai Dr. Manisha Singodia (MVSc Poultry Science, Jaipur) Dr. Annanda Das (Ph. D Scholar, WBUAFS, Kolkata) Dr. M. Arul Prakash (MVSc Assistant Professor, Tanjore) Dr. B.L. Saini (Ph. D ICAR, Izatnangar)

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DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

A WAY OF SCIENTIFIC MANAGEMENT OF FOOT AND MOUTH DISEASE (FMD) IN INDIAN DAIRY LIVESTOCK FARM Introduction

Mode of Transmission:

FMD is a highly infectious and an economically important viral disease of cloven- footed farm animals in India. The virus survives in lymph nodes and bone marrow at neutral pH, but is destroyed in muscle when pH is less than 6.0, i.e., after rigor mortis. The virus can persist in contaminated fodder and the environment for up to 1 month, depending on the temperature and pH conditions. It causes an estimated loss of Rs. 20-22 thousand crores per year to livestock owners in India. It mostly manifests the lesions in the mouth, feet and mammary gland. Milk yield drops dramatically in milking animals, suckling calf usually die and pregnant animals may abort and infertility may ensure following abortion. To control the disease, DAHDF of India launched a National FMD Control Program (FMD-CP) in 2003 with an outlay of about Rs. 500 crores a year by Central Government and each state government also invested an equally good amount of money.

FMD is a highly communicable contagious viral disease. FMD viruses can be spread by animals, people, or materials that bring the virus into physical contact with susceptible animals. 1. Generally by direct or indirect contacts between susceptible and infected animals. 2. Infected animals have a large amount of aerosol virus in their exhaled air, which can infect other animals via the respiratory or oral routes. 3. All secretions and excretions from the infected animal such as saliva, faeces and urine. 4. Through movement of clinically affected animals, animal handlers, visitors, physicians and inanimate vectors. 5. The disease has been transmitted to calves via infected milk.

1. Primarily affects cloven-hooved animals of the order Artiodactyla.

6. Virus can survive in dry fecal material for 14 days in summer, in slurry up to 6 months in winter, in urine for 39 days and on the soil between 3 (summer) and 28 days (winter).

2. Livestock hosts - cattle, pigs, sheep, goats, and experimental infections in alpacas and llamas.

7. By consumption of infected meat and meat by-products, unprocessed and uncooked milk

3. Repor ted in >70 species of wild artiodactyls, including bison, giraffes, Indian elephants, and several species of deer and antelope

8. Most of the animals remain as a carrier following recovery after infection.

Host Range:

Causes: 1. Caused by Aphthovirus of the family Picornaviridae 2. There are seven known serotypes: A, O, C, Asia 1, and SAT (Southern African Territories) 1, 2, and 3 3. More than 60 subtypes of the FMD virus 4. Immunity to one type does not protect an animal against other types 5. Six serotypes occur in Africa (O, A, C, SAT-1, SAT-2, SAT-3) 6. Four serotypes occur in Asia (O, A, C, Asia-1) and three serotypes occur in South America (O, A, C) 7. FMD in India is caused by FMDV Serotype O, A, & Asia 1 05

An outbreak can occur when: I. A n i m a l s c a r r y i n g t h e v i r u s a r e introduced into susceptible herds. ii. Contaminated facilities are used to hold susceptible animals.

vii. Susceptible animals drink common source contaminated water. viii.A susceptible animal is inseminated by semen from an infected animal. ix. Direct contact or aerosolized virus via respiratory secretions, milk, semen, and ingestion of feed from infected animals (meat, offal, milk). Symptoms: Signs of illness can appear after an incubation period of 1 to 8 days, but often develop within 3 days. 1. High fever up to 104-106˙F (41˙C) and anorexia. 2. Vesicles followed by erosions in the mouth, on the feet and in the mammary gland. 3. Vesicles that rupture and discharge clear or cloudy ﬂuid, leaving raw, eroded areas surrounded by ragged fragments of loose tissue. 4. Reduced consumption of feed due to painful tongue and mouth lesions. 5. Profuse salivation (saliva hanging in long ropy strings up to the ground). 6. Animal stamps its feet and wounds in the inter-digital space of legs followed by lameness 7. Oral ulcers, lesions and Smacking of lips. 8. Low milk production (dairy cows). 9. Myocarditis and death, especially in newborn animals. 10. Animals do not normally regain lost weight for many months. Disease Symptoms in the Mouth

iii. Contaminated vehicles are used to move susceptible animals. iv. Raw or improperly cooked garbage containing infected meat or animal products is fed to susceptible animals. v. People wearing contaminated clothes or footwear, or using contaminated equipment; pass the virus to susceptible animals. vi. Susceptible animals are exposed to materials such as hay, feedstuffs, hides, or biologics contaminated with the virus.

DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

5. Commercially available lateral ﬂow devices have not yet been validated by the OIE. Preventive and control measures: 1. We should provide most comfortable bedding (Sand) prior to calving and in advanced stage of pregnancy. 2. Early detection and treatment of milk fever. 3. Recumbent animals should be treated as soon as possible. 4. Cow should be bred with a bull as per its size as a big calf in a small cow will invite dystocia problem leading to calving paralysis. 5. Cow should not be made over fatty through too much feeding during advance pregnancy. Disease Symptoms in the Foot

6. Cow should be made to stand within a short time following parturition. 7. Parenteral Vitamin D3 should be given in milk fever prone cow during pregnant period.

FMD Control Treatment: 1. Permitted only in enzootic regions. 2. Only supportive type. 3. Control: I. Restricted movement of animals and vehicle movement around infected premises, proper carcass disposal, and environmental disinfection. II. Regions where FMD is not enzootic are usually controlled by slaughter of all infected and susceptible animals. III. Inactivated virus vaccines are limited in their use, because: Ÿ

They protect for only 4–6 mo against the speciﬁc serotype(s)

Protect animals from clinical illness but not viral persistence in the pharyngeal region; therefore, they can induce a carrier state

It is difﬁcult to distinguish infected animals from vaccinated animals unless puriﬁed killed vaccines are used

Thus, vaccination is used more in enzootic countries to protect producing animals, particularly high-yielding dairy cattle, from clinical illness because slaughter of all at-risk individuals may be legally/ economically unfeasible

8. Low calcium and high phosphorus diet should be given to stimulate parathyroid gland and thus to avoid hypocalcaemia. 9. If possible, cow of a dairy farm should be brought under metabolic proﬁle test to pinpoint the deﬁcit and to make good use of it. 10. Animal's both fore and hind limbs should be massaged two times per day.

Diagnosis: 1. In cattle, the clinical signs of FMD are indistinguishable from those of vesicular stomatitis. 2. L a b o r a t o r y d i a g n o s i s i s u s u a l l y performed via antigen capture–ELISA or serotyping ELISA. This is the preferred method for endemic FMD for viral antigen detection and serotyping. 3. Detecting nucleic acids via RT-PCR combined with real-time PCR is more sensitive and rapid than conventional methods and may be more useful when samples contain low concentrations of virus. 4. ELISA is preferred over complement ﬁxation tests because of its increased sensitivity and speciﬁcity.

11. Downer animals should be milked normally and the udder kept clean by washing with germicide soap before milking and post milking teat dips should be applied. 12. Re-placement therapy with Calcium, Phosphorus, magnesium, Glucose containing preparations can be used parentally by qualiﬁed veterinarian. 13. Infective causes should be brought under antibiotic coverage. 14. Physiotherapy by adopting muscle massage may be made to restore muscle activity of the limbs.

4. What we need to do for control of FMD? Ÿ

Rapid disease reporting system to control an FMD outbreak

Veterinarians who encounter any vesicular disease should immediately inform their regional veterinary authorities

After an outbreak, tracing must be done through epidemiologic inquiries to help identify the source of disease introduction

The dead infected carcasses must be disposed of via incineration, burial, or rendering on or close to the infected premises

People that have come into contact with virus may be asked to decontaminate their clothing and avoid contact with susceptible animals for a period of time.

B. L. Saini , S. Panda , M. Tarang , R. K. Jaiswal , A. Mehrotra , a a c a Soni Kumari , Adesh Kumar , S. K. Tanwar , and B. C. Naha a Ph. D. Scholar, Division of Animal Genetics, ICAR-IVRI, b Izatnagar, Ph. D. Scholar, Division of LPT, c Ph. D. Scholar, Division of P&C, ICAR-IVRI, Izatnagar

DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

LEPTOSPIROSIS - AN OVERVIEW Introduction

Source of infection

Leptospirosis is an anthrapozoonosis of ubiquitous distribution, caused by spirochetes of pathogenic Leptospira species. More than 250 pathogenic serovars of Leptospira are arranged in 25 serogroups. Leptospires are gram negative, ﬂexible helical rods that are actively motile, range from 10-20 µm in length with a diameter of 0.1-1.5 µm. They are too thin to be seen under light microscope and are best visualized under dark ground microscope. The disease is commonly found in humid tropical and subtropical areas. The magnitude of the problem in tropical and subtropical regions can be largely attributed to climatic and environmental conditions.

The most frequent sources of infection are urine, surface waters contaminated with urine of rats, animals humans, milk, abor ted fetuses and after bir th, reproductive discharges, mud and soil.

Epidemilogy

Reservoirs of Leptospira: Chronically infected carrrier animals (cattle, pigs, dogs rodents, small marsupials) The primary reservoir hosts for most Leptospira servars are wild mammals, particularly rodents. Reservoir hosts among domestic animals include cattle, pigs, sheep and dogs. The reservoir hosts vary with the serovar and the geographic region. Disease in reservoir hosts is more likely to be asymptomatic, mild or chronic. Reservoir hosts include. Ÿ

Rats: Serogroups i n t e ro h a e m o - 9 r r hagiae and ballum

Mice: Serogroup ballum

Cattle: Serovars hardjo, grippotyphosa and Pomona

Sheep: Serovars hardjo and Pomona

Pigs: Serovars pomana, tarassovi and Bratislava

Species Affected All mammals appear to be susceptible to at least one species of Leptospira. Disease is rare in cats, and less common in sheep than cattle. Ÿ

Serovars associated with disease in cattle include hardjo, pomana, grippotyphosa, canicola and icterohaemorrhagiae. Serovars associated with disease in sheep and goats include hardjo, pomana, grippotyphosa, and ballum. Serovars associated with disease in pigs include pomana, grippotyphosa, Bratislava, canicola, icterohaemorrhagiae, tarassovi and muenchen.

Serovars associated with disease in horses include hardjo, pomana, canicola, icterohaemorrhagiae, and sejroe.

Serovars associated with disease in dogs include pomana, grippotyphosa, canicola, icterohaemorrhagiae, pyrogens, paidjan, tarassovi, ballum and bratislava.

drainage fluids, drain water, mud and soils. Indirect transmission may be facilitated by water birds, arthropods, reptiles and worms. This risk group include veterinarians, rice field workers, slaughter house workers, butchers, laboratory workers, sewage workers, drainers, fisherman and agricultural workers especially sugarcane harvesting workers, miners and persons engaged in water sports. Leptospirosis situation in India The leptospirosis situation in India is a cause of concern. The endemicity of the disease is expanding to cover more states in the country. Geographic distribution of leptospirosis in the country is depicted in Table 1.

Table 1: Distribution of Leptospira servars in India

D o g s : S e ro va r s c a n i c o l a a n d bataviae

Direct transmission: Transplacental or congenital infection, sexual contact, suckling milk from an infected mother, artiﬁcial insemination with contaminated semen, invitro f e r t i l i z a t i o n w i t h c o n t a m i n a te d embryo, handling infected animals or animal products and from infected pet animals. Indirect transmission: Urine of excretor, carrier animals contaminated surface waters (ponds, lakes, rivers, streams), sewage, slaughter house

Based on isolation and serology Source: IVRI Izatnagar

Pathogenesis Leptospires enter through small cuts or aberrations or possibly through skin, or by inhalation of aerosols of urine. They spread immediately and circulate in the blood stream and diagnostically signiﬁcant bacteraemic phase lasts for about 1-7 days. After the numbers of leptospires in the blood and tissues reach a critical level, lesions due to a c t i o n of l e p to s p i r a l tox i n a n d consequent symtoms appear. The primar y lesion is damage to the endothelium of small blood vessels, leading to localized ischemia in various organs where the organisms localize DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

resulting in renal tubular necrosis, hepatocellular damage, meningitis, myositis and placentitis. Haemorrhages occur in severe cases as do jaundice and frequently, platelet deficiency. There is usually a mild granulocytosis and splenmegaly. Once immunity develops, leptospires are removed from the circulation and from tissues and organs by phagocytosis, following opsonization as seen by the appea-rance of circulating antibodies. Tissue damage, even though it is severe, may be reversible and followed by complete repair. However, long-lasting damage may be a complication and the clinically recovered animals may become chronic carrier, which act as source if infection to other animals and human beings. Clinical features and pathology Leptospirosis in domestic animals The serogroups reported in animal leptospirosis are presented in Table – 1 a. Bovines Bovine leptospirosis occurs worldwide. Leptospirosis in cattle ranges from totally inapparent to acute, febrile, and severe disease. Severity of illness seems to depend on the age and relative immunity of the animal, the infecting serovar, and the size and virulence of the dose. In its most flagrant form it is manifested by listlessness, loss of appetite, irritability, fever, ruffled fur, red eyes, and sometimes diarrhea, occurring 3-7 days after infection. There may be signs of haem orrhages and j aundi ce. Movement is accompanied by a characteristic arching of the back. Recovery may occur from here, or death may supervene. Recovery may be accompanied or followed by weight loss, runting in young animals, chronic renal failure and its signs and delayed death. In milk-producing cattle, there may be disturbance of milk flow and quality. Congenital infection of fetuses in utero follows a similar course, leading 09

to abortion if the fetus dies. The abortion products may be hemorrhagic or jaundiced or both; they may be heavily loaded with leptospires and a danger to animal handler. Still born fetuses may likewise be hemorrhagic jaundiced and infectious. Following abortion or stillbirth, the placenta is oedematous and the cotyledons are necrotic. Milk drop and agalactia, and discoloured milk are attributed to infections with serovar hardjo. Acute leptospirosis may present as a severe disease in calves and feeder cattle infected with a wide range of serovars, particularly, serovar Pomona. Clinical signs include high fever, haemolytic anaemia, haemoglobinuria, jaundice, congestion, and significant renal lesions. When pregnant cattle are infected with these serovars abortion storms can result. In lactating cows, incidental host infections are often associated with agalactia, with small quantities of blood-tinged milk . Recovery is prolonged. The most common clinical syndrome associated with acute leptospirosis occurs in dairy cows as a transient fever with a precipitous drop in milk production lasting for two to ten days. Leptospiral milk drop syndrome varies from an epizootic infection in a previously unexposed herd, involving over half the herd over a period of one or two months, to a more common, endemic infection affecting cows in their first or second lactation. Recovery is usually within 10 days although cows that experience a significant drop in milk production during the lactation period. Animals which have recovered from acute leptospirosis may develop a carrier condition in which leptospires grow and may remain in the renal tubules, for periods of days to years. From here they are passed out in the urine. These excretor animals are the central points of distribution for

leptospirosis in other animals or people. Leptospires may also persist in other organs, notably the genital tract. The chronic form of bovine leptospirosis is associated with fetal infection in pregnant cows presenting as abortion, stillbirth or birth of premature and weak infected calves but apparently healthy calves also may be born. Retention of fetal membranes may follow hardjo abortion. Congenitally infected new born calves are often weak , and affected by degeneration of the liver or kidneys or both. They are prone to succumb to secondary infection. There are sub acute and chronic kidney lesions, seen macroscopically as focal white spots and sub capsular scarring on the surface of the kidneys develop in chronically infected cattle regardless of age. Specimen to be collected Acute leptospirosis: Live animals; Blood, CSF, urine, peritoneal or pleural exudates collected during first 5-7 days for detection and isolation of leptospira. Dead animals: Kidney, liver, brain and anterior chamber of eye Abortion: Fetal kidney, liver, placenta and uterine discharges Chronic leptospirosis: Brain, genital tract discharges, kidney from dead animals Live animals: Urine, CSF and kidney biopsy Diagnosis 1. Direct examination of blood, body fluids and tissues: During the first week to 10 days, leptospires may be seen on direct mocroscopic examination of blood, peritoneal or pleural exudates and urine. The advantage of direct observation is speed. The disadvantages are the technical difficulties of obtaining suitable specimens, the requirement of skilled personnel and inability to differentiate from non-pathogenic leptospires. DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

2. Culture of leptospires from clinical specimens, blood, CSF, urine and tissues samples collected under aseptic conditions are inoculated into leptospires growth medium containing antibiotics and 5 flurouracil. Bind subcultures after one day incubation can improve chances for isolation. Addition of rabbit serum may enhance the growth of more fastidious leptospires. 3. Molecular approaches PCR with specific primers targeting 16S RNA detected pathogenic leptospires in many biological specimens including urine, CSF, and aqueous humor. PCR have advantages of speed, sensitivity and specificity. The limitations are the need of special equipment, the relatively high cost of the reagents, absence of automated and standardized procedures allowing the testing of large sets of samples, particularly in tropical countries where the disease is endemic. 4. Serological diagnosis Microscopic agglutination test (MAT), IgM ELISA, Dot-ELISA, Lateral flow kits, latex agglutination kits, and microcapsule kits are available for diagnosis of leptospirosis. Blood for serology should be taken as soon as possible in the illness. A second specimen should be taken 5-7 days later and repeated at similar intervals if necessary. Antibody levels drop over weeks or months but may persist for 2-10 years. In endemic areas, where many people or animals may have had leptospirosis, the interpretation of a single low titre may be impossible; a rising titre in successive specimens is then mandatory for serological diagnosis. Where there are antibodies to several serovars 10

detected in early sera, the highest titre may not be against the infecting serovar. The highest titre in later sera tends to be more serovar specific. IgM antibodies produced early in infection can be detected with specific anti-IgM ELISA and is more promising for detection of early leptospirosis. IgM ELISA in dipstick form is more appealing due to its ease of use, compatible with field conditions encountered where there are few medical resources. This type of test is essentially useful only for screening as it is limited by the same constraints that apply to conventional ELISA methods. In epidemiological studies the data provided by this type of method need to be validated using MAT in parallel as a reference method. MAT is specific for the infecting serovar or closely antigenically related serovars. Most laboratories use at least 6, up to 15 serovars, representative the locally common serogroups. The MAT has the disadvantages that it is tedious and wasteful to test against a large battery of serovars to ensure that the infecting serovar is included. Slide agglutination test, immune haemagg-lutination and complement fixation and microcapsule agglutination tests are less tedious and but are specific and sensitive than MAT. A general recommended standard is that, in an endemic area, a serological diagnosis of leptospirosis is confirmed in human patients or animals with a compatible clinical illness, where the

microscopic agglutination titre in sera taken 5-10 days apart rises from less than the starting dilution of the test or more than 100, or 4-fold or more, or is initially more than 400 in a nonendemic area, a single titre of 50 or more, with a clinically compatible illness, indicates likely leptospirosis, the titre will usually rise 4-fold or more in a second specimen a few days later. However, a single titre of 400 or more can lead to only presumptive diagnosis, indicating recent contact with leptospires which are not necessarily the cause of current illness. The titre to some serovars is consistently higher than to others. Consistent criteria are required for the definition of presumptive and confirmed cases using serological. These two categories need to be separated, otherwise a diagnosis is subjective and epidemiological patterns cannot be validly compared. Treatment Antibiotic treatment is effective within the first 7 to 10 days of infection. It should be given immediately on diagnosis or suspicion. Rapid effective antibiotic treatment can reduce the a d va n c e of s y m p t o m s a n d t h e likelihood of sequelae. It can thus prevent progress to the severe manifestations of illness. Antibiotics will eliminate leptospires from accessible tissues but will not reverse already established pathological changes, which must run their course and be treated symptomatically or by appropriate non-antibiotic therapy.

K. Manimaran*, S. Balakrishnan, A. Sangeetha and M. Dhanalakshmi Department of Veterinary Public Health and Epidemiology Veterinary College and Research Institute, Orathanadu-614 625,Thanjavur Dt.

DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

ETIOPATHOGENESIS AND DIFFERENT APPROACHES FOR BOVINE KETOSIS Introduction Ketosis is a multifactorial disorder of energy metabolism. Negative energy results in hypoglycemia and ketonemia (the accumulation in blood of acetoacetate, B-hydroxybutyrate and their decarboxylation products acetone and isopropanol). Etiology Glucose metabolism in ruminants The maintenance of adequate concentrations of glucose in the blood is critical to the regulation of energy metabolism. The ruminant absorbs very little dietary carbohydrate as hexose sugar because dietary carbohydrates are fermented in the rumen to short chain fatty acids, principally acetate (70%), propionate (20%) and butyrate (10 %). Propionate and amino acids are the major precursors for gluconeo-genesis with glycerol and lactate of lesser importance. Propionate is produced in the rumen from starch, fiber, and proteins. It enters the portal circulation and is efficiently removed by the liver, which is the primary glucose producing organ. Amino acids The majority of amino acids are glucogenic and are also important precursors for gluconeogenesis. Dietary protein is the most impor tant quantitative source but the labile pool of body protein is also an important source; together they contribute to energy synthesis and milk lactose synthesis as well as milk protein synthesis. Energy balance In high-producing dairy cows there is often a negative energy balance in the first few weeks of lactation. The highest dry matter intake does not occur until 8-10 weeks after calving but peak milk production is at 4-6 weeks and energy intake may not keep up with demand. In response to a negative energy balance and low serum concentrations of glucose and insulin, cows will mobilize adipose tissue 11

with consequent increases in serum concentrations of non-esterified fatty acids (NEFA) and subsequently BHBA. Hepatic in sufficiency in ketosis Hepatic insufficiency has been shown to occur in bovine ketosis. It has been suggested that ketosis can be divided into two types: I) In Type I, or 'spontaneous' ketosis it is proposed that the gluconeogenic pathways are maximally stimulated. Rapid entry of non-esterified fatty acids (NEFA) into hepatic mitochondria occurs and results in high rates of ketogenesis and high blood ketones. II) In Type II ketosis, manifest with fatty liver, gluconeogenic pathways are not maximally stimulated and consequently mitochondrial uptake of NEFA is not as active and NEFA become esterified in the cytosol, forming triglyceride. The capacity of cattle to transport triglyceride from the liver is low, resulting in accumulation and fatty liver. Ketone formation Ketones arise from two major sources: butyrate in the rumen and mobilization of fat. A large proportion of butyrate produced by rumen fermentation of the diet is converted to 13-hydroxybutyrate (BHBA) in the rumen epithelium and is absorbed as such. Free fatty acids produced from the mobilization of fat are transported to the liver and oxidized to produce acetyl-CoA and NADH. AcetylCoA may be oxidized via the TCA cycle or metabolized to acetoacetyl CoA. Its oxidation via the TCA cycle depends upon adequate supply of oxaloacetate from the precursor propionate. I f p ro p i o n a te , a n d c o n s e q u e n t l y oxaloacetate, is deficient, oxidation of acetyl-CoA via the TCA cycle is limited and it is metabolized to acetoacetyl CoA and subsequently to acetoacetate and BHBA. Role of insulin and glucagon The regulation of energy metabolism in

ruminants is primarily governed by insulin and glucagon. Insulin acts as a glucoregulatory hormone stimulating glucose use by tissues and decreasing hepatic gluconeogenesis. Blood insulin concentrations decrease with decreasing blood concentrations of glucose and propionic acid. Insulin also acts as a liporegulatory honnone stimulating lipogenesis and inhibiting lipolysis. Glucagon is the primar y counterregulatory hormone to insulin. A low insulin: glucagon ratio stimulates lipolysis in adipose tissue and ketogenesis in the liver. Elevated ketones may stimulate insulin production and may act as a negative feedback. Regulation is also indirectly governed by somatotropin, which is the most important determinant of milk yield in cattle and is also lipolytic. Etiology of bovine ketosis High-yielding cows in early lactation are in negative energy balance and are subclinically ketotic as a result. This can be predisposed by nutrition inadequacies during the dry period. Ruminants are particularly vulnerable to ketosis because, although very little carbohydrate is absorbed as such, a direct supply of glucose is essential for tissue metabolism, particularly the formation of lactose. The utilization of volatile fatty acids for energy purposes is also dependent upon a supply of available glucose. In the period between calving and peak lactation, the demand for glucose is increased and cannot be completely restrained. Cows will reduce milk production in response to a reduction of energy intake, but this does not follow automatically nor proportionately in early lactation because hormonal stimuli for milk production overcome the effects of reduced food intake. Under these circumstances, lowered blood glucose levels result in a lowered blood insulin. Individual cow variation The rate of occurrence of negative energy DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

status, and therefore the frequency of clinical cases, has undoubtedly increased sharply in the recent past because of the steep increase in the lactation potential of the modern dairy cow. Because of the mammary gland's metabolic precedence in the partitioning of nutrients, especially glucose, milk production continues at a high rate, causing an energy drain. Classification Lean has presented a classification of the disease based on its natural presentation in intensively and extensively managed dairy herds, and one that accounts for the early lactational demand for glucose, a limited supply of propionate precursors and preformed ketones or mobilized lipids in the pathogenesis. Such a classification includes the following geneses of ketosis, which will be discussed in turn: Ÿ

Primary ketosis (production ketosis)

Secondary ketosis

Alimentary ketosis

Starvation ketosis

Ketosis due to speciﬁc nutritional deﬁciency.

Primary ketosis (production ketosis) This is the ketosis of most herds, the so called estate acetonemia. It occurs in cows in good to excessive body condition that have high lactation potential and are being fed good-quality rations but that are in a negative energy balance. A proportion of cases appear as clinical ketosis but a much greater proportion occur as cases of subclinical ketosis in which there are increased levels of circulating ketone bodies but no overt clinical signs. Secondary ketosis This occurs where other disease results in a decreased food intake. The cause of the reduction in food intake is commonly the result of abomasal displacement, traumatic reticulitis, metritis, mastitis, or other diseases common to the postparturient period. A high incidence of ketosis has also been observed in herds affected with ﬂuorosis. An unusual occurrence reported was an outbreak of 12

acetonemia in a dairy herd fed on a ration contaminated by a low level (9.5 ppm) of lincomycin, which caused ruminal microbial dysfunction. Alimentary ketosis This form is due to excessive amounts of butyrate in silage and possibly also due to decreased food intake resulting from poor palatability of high butyrate silage. Silage made from succulent material may be more highly ketogenic than other types of ensilage because of its higher content of preformed butyric acid. Spoiled silage is also a cause and toxic biogenic amines in silage, such as putrescine, may also contribute.This type of ketosis is commonly subclinical but it may predispose to the development of production or primary ketosis. Starvation ketosis This occurs in cattle that are in poor body condition and that are fed poor-quality feedstuffs. There is a deficiency of propionate and protein from the diet and a limited capacity of gluconeo-genesis from body reserves. Affected cattle recover with correct feeding. Ketosis due to specific nutritional deficiency Specific dietary deficiencies of cobalt and possibly phosphorus may also lead to a high incidence of ketosis. This may be due in part to a reduction in the intake of total digestible nutrients (TDN), but in cobalt deficiency, the essential defect is a failure to metabolize propionic acid into the tricarboxylic acid (TCA) cycle. There is a marked nadir in food intake around calving, followed by a gradual increase. This increase is quite variable between cows, but in the great majority of cases does not keep pace with milk yield. Epidemiology Occurrence Ketosis is a disease of dairy cattle and is prevalent in most countries where intensive farming is practiced. It occurs mainly in animals housed during the winter and spring months and is rare in cows that calve on pasture. In housed or

free-stalled cattle it occurs year around. The occurrence of the disease is very much dependent upon management and nutrition and varies between herds. Lactation incidence rate for ketosis that va r i e d f ro m 0 . 2 - 1 0 . 0 % . R a t e s of subclinical ketosis are influenced by the cut-point of plasma BHBA used for definition but are much higher, especially in undernourished herds and can approach 40%. Animal and management risk factors There are conflicting reports on the significance of risk factors for ketosis and subclinical ketosis which probably reflect that the disease can be a cause or effect of interacting factors. The disease occurs in the immediate postparturient period with 90% of cases occurring in the first 60 days of lactation. Regardless of specific etiology, it occurs most commonly during the first month of lactation, less commonly in the second month, and only occasionally in late pregnancy. Age Cows of any age may be affected but the disease increases from a low prevalence at the first calving to a peak at the fourth. Lactational incidence rates of clinical ketosis of 1.5% and 9%, respectively were found in a study of 2415 primiparous and 4360 multiparous cows. Clinical ketosis can also recur in the same lactation. Herd differences in prevalence are very evident in clinical practice, and in the literature, with some herds having n e g l i g i b l e o c c u r re n c e . A l t h o u g h apparent differences in breed incidence are reported, evidence for an heritable predisposition within breeds is minimal. Body condition score (BCS) There are conflicting reports on the relation between BCS at calving and ketosis but it is suggested that studies that have found no relationship have not had many fat cows in the herds examined. Fat body condition post partum was observed to be associated with a higher first test day milk yield, milk fat to protein ratio of > 1.5, increased body condition loss and a higher risk for ketosis. Body condition loss during the DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

dry period also increases risk for ketosis in the following lactation.

the experimental, IV or SC injection of insulin (2 units /kg BW).

Season

However, in most field cases the severity of the clinical syndrome is also roughly proportional to the degree of ketonemia. This is an understandable relationship as ketone bodies are produced in larger quantities as the deficiency of glucose increases. However, the ketone bodies may exert an additional influence on the signs observed.

There is no clear association with season. In some but not all summer grazing areas, a higher risk is generally observed in cattle during the winter housing period. Higher prevalence has been observed in the late summer and early winter in Scandinavian countries. Other interactions There is a greater risk for the development of ketosis in cows that have an extended long dry period, that develop milk fever, retained placenta, lameness or hypomagnesemia. Cows with twins are also at risk for ketosis in the terminal stages of pregnancy. Economic signiﬁcance Clinical and subclinical ketosis are major causes of loss to the dairy farmer. In rare instances the disease is irreversible and the affected animal dies but the main economic loss is due to the loss of production while the disease is present, the possible failure to return to full production after recovery and the increased occurrence of periparturient disease. Both clinical and subclinical ketosis are accompanied by decreased milk yields and lower milk protein and milk lactose and increased risk for delayed estrus and lower ﬁrst service conception rates, increased inter-calving intervalsl and increased risk of cystic ovarian disease, metritis and mastitis and increased involuntary. Pathogenesis The principal metabolic disturbances observed, hypoglycemia and ketonemia, may both exert an effect on the clinical syndrome. In many cases, the severity of the clinical syndrome is proportional to the degree of hypoglycemia and this, together with the rapid response to parenterally administered glucose in cattle, suggests hypoglycemia as the predominant factor. This hypothesis is supported by the development of prolonged hypoglycemia and a similar clinical syndrome to that of ketosis, after

The nervous signs which occur in some cases of bovine ketosis are thought to be caused by the production of isopropyl alcohol, a breakdown product of acetoacetic acid in the rumen, although the requirement of nervous tissue for glucose to maintain normal function may also be a factor in these cases. Spontaneous ketosis in cattle is usually readily reversible by treatment; incomplete or temporary response is usually due to the existence of a primary disease with ketosis present only as a secondary development, although fatty degeneration of the liver in protracted cases may prolong the recovery period. Immunosuppression has been demonstrated with energy deficiency and ketosis. The higher susceptibility of ketotic postpartum cows to local and systemic infections may be related to impairment of the respiratory burst of neutrophils which occurs with elevated levels of BHBA. Clinical findings Two major clinical forms: wasting and nervous, but these are the two extremes of a range of syndromes in which wasting and nervous signs are present in varying degrees of prominence. The wasting form is the most common of the two, manifest with a gradual but moderate decrease in appetite and milk yield over 2-4 days. In herds that feed components separately the pattern of appetite loss is often unusual in that the cow first refuses to eat grain, then ensilage but may continue to eat hay. Body weight is lost rapidly, usually at a greater rate than one would expect from the decrease in appetite. Farmers usually

describe affected cows as having a 'woody' appearance due to the apparent wasting and loss of cutaneous elasticity due presumably to disappearance of subcutaneous fat. The temperature and the pulse and respirator y rates are normal and although the ruminal movements may be decreased in amplitude and number, they are within the normal range unless the course is of long duration when they may virtually disappear. A characteristic odor of ketones is detectable on the breath and often in the milk. In the wasting form, nervous signs may occur in a few cases but rarely comprise more than transient b o u t s of s t a g g e r i n g a n d pa r t i a l blindness. In nervous form, signs are usually bizarre and begin quite suddenly. The syndrome is suggestive of delirium rather than of frenzy and the characteristic signs include: Walking in circles, straddling or crossing of the legs, head pushing or leaning into the stanchion. Apparent blindness Aimless movements and wandering vigorous licking of the skin and inanimate objects depraved appetite and chewing movements with salivation. Hyperesthesia may be evident, the animal bellowing on being pinched or stroked. Subclinical ketosis Many cows that are in negative energy balance in early pregnancy will have ketonuria without showing clinical signs, but will have diminished productivity inciuding depression of milk yield and a reduction in fertility. Infertility may appear as an ovarian abnormality, delayed onset of estrus or as endometritis resulting in an increase in calving to conception interval and re d u c e d c o n c e p t i o n r a te a t ﬁ r s t insemination. Clinical Pathology Hypoglycemia, ketonemia and ketonuria are characteristic of the disease. Glucose : Blood glucose levels are reduced from the normal of approximately 50 mg/dL to 20-40 mg/dL. DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

Ketosis secondary to other diseases is usually accompanied by blood glucose levels above 40 mg/dL and often above normal. Blood ketones : Plasma or serum hydroxybutyrate (BHBA) measured in SI units is used for analysis of ketonemia. BHBA is the predominant Circulating ketone body. Normal cows have plasma BHBA concentrations less than 1000 mol/L, cows with subclinical ketosis have concentrations greater than 1400 mol/L, and cows with clinical ketosis have concentrations often in excess of 2500 mol/L. Milk and urine cowside tests Cowside tests have the advantage of being inexpensive, giving immediate results, and they can be used as frequently as necessary. A minor source of error is that the concentration of ketone bodies in these fluids will depend not only on the ketone level of the blood but also on the amount of urine excreted or on the milk yield. Milk is less variable, easier to collect and may give fewer false negatives with subclinical ketosis. Milk and urine ketone levels have been traditionally detected by the reaction of acetone and acetoacetate with sodium nitroprusside and can be interpreted in a semi-quantitative manner based on the intensity of the reaction. The sensitivity and specificity of the nitroprusside powder test with milk in various studies is reported as 28-90% and 96-100%, respectively. Urine testing A nitroprusside tablet has a reported sensitivity and specificity of 100% and 59%, respectively. Milk fat concentration tends to increase and milk protein concentration tends to decrease during postpartum negative energy balance. A fat to protein ratio >1.5 in first day teat milk is indicative of a lack of energy supply in the feed and of risk for ketosis. Clinical chemistry and hematology White and differential cell counts are variable and not of diagnostic value for ketosis. There are usually elevations of liver enzymes but liver function tests are within the normal range. Liver biopsy is the only accurate method to determine 15

the degree of liver damage. Plasma concentrations of non-esterified fatty acids are elevated as are cholesterol concentrations and bilimbin. Bilirubin is not a sufficiently sensitive indicator to asses the extent of fat mobilization and liver function. Necropsy findings The disease is not usually fatal in cattle but fatty degeneration of the liver and secondary changes in the anterior pituitary gland and adrenal cortex may be present. Differential diagnosis Cattle The clinical picture is usually too indefinite, especially in cattle, to enable a diagnosis to be made solely on clinical grounds. General consideration of the history, with particular reference to the t i m e of c a l v i n g , t h e d u r a t i o n of pregnancy in ewes and the feeding program, and biochemical examination to detect the presence of hypog-lycemia, ketonemia, and ketonuria are necessary to establish a diagnosis. Wasting form Abomasal displacement Traumatic reticulitis Primary indigestion Cystitis and pyelonephritis Diabetes mellitus Nervous form Rabies Hypomagnesemia Bovine spongiform encephalopathy Treatment I n c a t t l e , a n u m b e r of e f f e c t i v e treatments are available but in some affected animals, the response is only transient; in rare cases, the disease may persist and cause death or necessitate slaughter of the animals. The simplest means of doing this is by the administration of glucose replacement therapy. The effect of the adminis-tration of glucose is complex but it allows the re v e r s a l of ke t o g e n e s i s a n d t h e establishment of normal patterns of energy metabolism.12 Replacement therapy Glucose: The injection of 500 mL of a 50% solution of glucose results in transient hyperglycemia, increased insulin and decreased glucagon secretion, and

reduced plasma concentration of non esteriﬁed fatty acids. It effects a marked improvement in most cows but relapses occur commonly unless repeated treatments are used. IP injections of 20% solution of dextrose may be used alternatively but are also accompanied by risk of infection. Propylene glycol and glycerine/glycerol To overcome the necessity for repeated injections, propylene glycol can be administered as a drench. The traditional does is 225 g twice daily for 2 days, followed by 110 g daily for 2 days to cattle, but higher volumes are also used. Propylene glycol (200-700 g daily), or salts of propionic acid, can be administered in the feed and give good results. Administration in feed is preferred by some because this method avoids dangers of aspiration with drenching; however, cows not used to its inclusion in the feed may show feed refusal. Other glucose precursors Because of its glucogenic effect, sodium propionate is theoretically a suitable treatment but when administered in 110225 g doses daily, the response in cattle is often very slow. Lactates are also highly glucogenic but both calcium and sodium lactate (1 kg initially, followed by 0.5 kg for 7 days) and sodium acetate (110-500 g/d) have given less satisfactory results than those obtained with sodium propionate. Hormonal therapy Glucocorticoids : Hyperglycemia occurs within 24 h of administration and appears to result from a repartitioning of glucose in the body rather than from gluconeogenesis. A hyperglycemic state is produced for 4-6 days in ketotic cows given 10 mg of dexamethasone 21isonicotinate and other preparations such as dexamethasone sodium phosphate (40 mg) and ﬂumethasone (5 mg) are also used Response of cows with primary ketosis to treatment with corticosteroids and IV glucose is superior, with fewer relapses, than therapy with corticosteroids or glucose alone. Insulin facilitates cellular uptake of glucose, suppresses fatty acid DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

metabolism and stimulates hepatic gluconeogenesis. The dose of protamine zinc insulin is 200-300 IU per animal administered SC every 24-48 h as required. Anabolic steroids have also been used for treatment of lactational ketosis and ketosis in late pregnant cows that are overfat, stressed, or have twin fetuses. Miscellaneous treatments: Vitamin B12 and cobalt are indicated in regions where cobalt deficiency is a risk factor f o r ke to s i s . T h e y a re s o m e t i m e s administered to cattle with ketosis in regions where cobalt deficiency does not occur but their therapeutic value is not proven. Control The control of clinical ketosis is integrally related to the adequate nutrition of the cow in the dry and lactating period. This encompasses details such as: Dry matter intake, Fiber digestibility, Par ticle size distribution, Energy density, Fat incorporation in early lactation rations Protein content, Feeding systems, Rumen size. It is difficult to make general recommendations for the control of the disease because of the many

conditions under which it occurs, its p ro ba b l e m u l t i p l e e t i o l o g y, a n d feeding systems that vary from those that feed components separately to those that feed total mixed rations. Cows should neither have been starved nor be overfat at calving. Too low a feeding frequency and the feeding of concentrates separate from roughage rather than as a total mixed ration can lead to an increase in rates of ketosis. In high-producing cows being fed stored feeds, poor quality roughage commonly leads to acetonemia. Wet ensilage containing much butyrate, and moldy or old and dusty hay, are the main offenders. Energy supplements Propylene glycol is used for the prevention of clinical and subclinical ketosis. Propylene glycol can be added to feed and is frequently present in

commercial feed product but a bolus dose of propylene glycol is more effective in raising blood glucose than incorporation in feed. lonophores Ionophores alter bacterial flora of the rumen, leading to decreases in Grampositive bacteria, protozoa, and fungi and increases in Gram-negative bacteria. The net effect of these changes in bacterial flora is increased propionate production and a decrease in acetate and butyrate production providing increased gluconeogenic precursors. Niacin Niacin is antilipolytic and induces increases in blood glucose and insulin but there is conflicting evidence that niacin given in the feed has a beneficial effect on subclinical ketosis in cattle.

Pankaj Kumar Patel¹, Sonam Bhatt¹, Anjali Singh² and Vandita Mishra³ ¹Division of Medicine, ²Physiology & Climatology, 3 Division of Livestock Product Technology, ICAR-IVRI, Izatnagar, U.P., India

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MASTITIS IN DAIRY : PREVENTION CAN SAVE COSTLY TREATMENT Mastitis (Greek, Mastos =breast + it is = inflammation) is a multietiological complex disease, which is defined as inflammation of parenchyma of mammary glands. Holstein Friesian (HF), Jersey and crossbred dairy cows are generally more susceptible to mastitis than indigenous breeds. Mastitis occurs throughout the world wherever dairy cows are found, generally higher in high yielding bovines. The disease may be attributed to deficient management, improper milking procedures, faulty milking equipment, inadequate housing. The most expensive disease on dairy farms is mastitis which includes cost of antibiotics,NSAID, in addition to loss of milk production.

Mastitis causes loss of 2500 to 5000 Rupees to a dairy farmer in one lactation….. Breed

Yield Discarded Production Treatment Replacement Total loss(Rs) milk(Rs) loss(Rs) Cost (Rs) (Rs) loss (Rs) ND 316 26 343 525 1580 2448 CB 546 72 618 695 2235 3549 Buffaloes 552 662 72 625 647 1934 Singh, D., Kumar, S., Singh, B. and Bardhan, D., 2014. Economic losses due to important diseases of bovines in central India. Veterinary World, 7(8).

Simple answer to mastitis (Prevention better than cure)

Retention time of the antimicrobial in the udder is longer.

200 microbial species of bacteria,virus, fungi, yeast, algae and chlamydia can cause mastitis in cattle and buffeloes. In India, Staphylococcus, Streptococcus and E.coli generally cause 90-95% of all infections. Effective and economical mastitis control programs rely on prevention rather than treatment.

The incidence of new infections during the dry period is reduced.

Tissue damage by mastitis may be regenerated before parturition.

Clinical mastitis at calving may be reduced.

The risk of contaminating milk with antimicrobial residue is reduced

Dry Cow Therapy the Ultimate and easiest answer to this complex problem D r y c o w t h e r a p y i s t h e u s e of intramammary antimicrobial therapy immediately after the last milking of lactation and is an important component of an effective mastitis control program. Intramammar y infusions at drying off decrease the number of existing infections and prevent new infections during the early weeks of the dry period Dry cow therapy has the following advantages: The cure rate is higher than that achieved by treatment during lactation. Ø

A much higher dose of antimicrobial can be used safely.

Sensitivity of important mastitis c a u s i n g o rg a n i s m s to va r i o u s antibiotics

Ondiek, J.O., Ogore, P.B., Shakala, E.K. and Kaburu, G.M., 2013. Prevalence of bovine mastitis, its therapeutics and control in Tatton Agriculture Park, Egerton University, Njoro District of Kenya. Basic Research Journal of Agricultural Science and Review, 2(1), pp.15-20

Giving an intramammary antibiotic in all teats during period when cattle is not in lactation is Dry Cow Therapy!!!

During the dry period i.e. during 60 days before giving birth to calf animal should be given a rest period

During this period intramammary i n j e c t i o n of C e p h a l e x i n a n d Neomycin preparation can be given for prevention of mastitis

For treatment of chronic cases a combination of antibiotics comprising Amoxicillin, Cephalexin, Neomycin and Enroﬂoxacin can be given for maximum efﬁcacy. Seema Yadav¹, PramodkumarSoni¹ and Kundan Kumar³ ICAR- Indian Veterinary Research Institute, Bareilly, U.P

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SNP CHIP FOR INDIGENOUS CATTLE AND BUFFALO Introduction The total population of Cattle, buffalo & goat in India is 190.9 million, 108.7 million and 135.17 million animals (19th livestock census). India is the largest producer of milk in the world with a total production of 165.4 million tonnes during 2016-17 (BAHS, 2017). The world’s total buffalo population is around 193.8 million, out which India (108.7 million) and Pakistan (33.6 million) accounts for 73.4% of the total world’s buffalo population (http:// faostat3.fao.org/) and is steadily increasing at the rate of 2% per annum during the last two decades. Of the total milk production, Cattle, buffalo & goat contribute a total of 46.2%, 49.2% and 4.6% respectively (BAHS, 2017). Even though India stands ﬁrst in terms of milk production in the world, the productivity per animal is very low and is 2.84 and 5.23 kg per day in Indian cattle and buffaloes, respectively and is very less when compared to the productivity per animal in the developed countries. The major causes of low productivity in India are both intrinsic (low genetic potential) and extrinsic (poor nutrition/feed management, inferior farm management practices, ineffective veterinary and extension ser vices and inefﬁcient implementation of breed improvement p r o g r a m m e s ) . I n c a t t l e m o s t of indigenous breeds are draught type and have not been selected for milk for centuries. Improvement of the milk production of indigenous bovines can be achieved by giving emphasis on the selection of individuals for use in the breeding programs. In the traditional selection program, the breeding value of animal is estimated based upon the information from its related individuals. Moreover, in traditional selection the selection of individuals depends upon availability of phenotypic observations, which further depends upon the heritability of the trait. For traits with low heritability, traditional selection may not be possible and it may 18

not provide a clear picture of the value of the animal and this method of selection is difficult to follow for the sex limited traits, traits that are expressed late in the animal's life and for the traits that cannot be measured easily. Modern breeding techniques in animal and plant species has been revolutionized by SNP arrays which helps to predict the genetic merit known as genomic selection.The term genomic selection was first introduced by Haley and Visscher in 1998, whereas the fundamental concept of GS was first put forward by Meuwissen et al. (2001). Presently, genomic selection is being practiced in many developed countries, viz., Australia, New Zealand, United States, Norway, Ireland, European union etc. The genomic information of the individual may help in the selection of individuals and also helps to predict the total genomic breeding values based upon a very large number of marker haplotypes across the entire genome. SNP chips are mainly used for ranking of sires for various traits, viz., Protein yield, protein percentage, fertility, Milk production traits, milk composition, fat yield, live Body weight, fertility, Somatic cell count, longevity, female fertility, feed efficiency, post-weaning growth, to estimate breed composition, first lactation mastitis traits, and calving ease, Bovine Tuberculosis Susceptibility, etc.

SNP chips allow analysis of many SNPs in parallel, which is essential for large scale association studies for quantitative traits in cattle. There are various SNP genotyping platforms, such as oligonucleotide arrays (Affymetrix, Inc., Santa Clara, CA, USA) and BeadArray microarrays (Illumina, Inc., San Diego, CA, USA). P re s e n t l y A f f y m e t r i x of f e r s S N P genotyping arrays for livestock and aquaculture species (buffalo, cattle, chicken, pig, salmon and trout), crops (cotton, maize, soybean, strawberry and wheat) and biomedical and model organisms (human, dog, mouse and Arabidopsis thaliana) (http://www.affymetrix.com), while Illumina is marketing wholegenome genotyping Bead Arrays for human and non-human species (cattle, dog, maize, pig and sheep) (http://www.illumina.com). SNP chip for cattle and buffalo cattle Reduction in cost of SNP genotyping and its accuracy in predicting breeding value using a dense markers made a revolutionar y change by reducing generation interval in dairy and beef bovine breeding. Buffalo

Presently, SNP chips are being used in various species and are being used mainly in Cattle and Buffaloes. The cattle SNP chips that’s being in use is biased towards the Bos taurus and this article focus upon the usability of available SNP chips in Indian cattle breeds for its use in predicting the genomic breeding value and genomic selection. SNP chip SNP chip is a small piece of silicon glass (~1 cm2) to which a large number of synthetic, single stranded DNA oligonucleotides are chemically bonded. The oligonucleotides function as DNA probes that anneal with only complementary DNA molecules. The DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

Table 1: SNP chips available in market for bovines Company

SNP chips

No. of SNPs

Name of Bos indicus No. of breeds Included Polymorphic loci

Illumina

Bovine HD777k

7,77,962

Brahman

5,61,834

Gir

4,72,928

Nellore Brahman

453361 33,038

Gir

25,320

Nellore

22,422

Bovine 50k

53,714

Bovine LD v2.0

7,931

GoldenGate Bovine 3k

2,900

Gene seek GGP 35Ki

35,090

GGP 150K

1,50,000

GGP F250

2,30,000

Affymetrix Axiom geomewide bos1 bovine array kit Axiom Bovine genotyping array

Brahman

6,48,855

Brahman Gir Nelore

6,48,855

The Illumina BovineSNP50 BeadChip was used to validate the DNA samples from three breeds of water buffalo (Nili-Ravi, Murrah and their crossbred with local GuangXi buffaloes in China). A total of 40,766 bovine SNPs were validated and found in the water buffalo genome. Of which, 935 were identiﬁed to be polymorphic and useful for association analysis in water buffalo. The SNP chip for buffalo population was developed using genomic information of several buffalo breeds (Italian Mediterranean, Murrah, Nili- Ravi, Jaffarabadi, Kundhi, Aza-Kheli, Egyptiana and Swamp type from Philippines) that facilitated the identiﬁcation of millions of sequence variants in the buffalo genomes. The buffalo genotyping array contains a total of 90k putative single nucleotide polymorphisms (SNP) and tested in buffalo populations from Italy and Brazil.This chip has been used to genotype river buffalo samples from Pakistan, Iran, Turkey, Egypt, Romania, Bulgaria, Italy, Mozambique, Brazil and Colombia, and swamp buffaloes from China, Thailand, Philippines, Indonesia and Brazil. SNP chip for Bos indicus and Bubalus bubalis The optimum number of SNPs for inclusion in the SNP chip depends upon the genome size and LD. After checking for LD, tagged SNPs may be considered for inclusion in chip. Maximum number of samples from different breeds of a species should be included. The rare alleles need to be excluded. 19

2,40,000 2,10,000 2,30,000

Disadvantage of using available SNP chips in Bos indicus The SNP chips that has been designed for Bos taurus breeds and validated in 19 common dairy and beef cattle breeds may not be suitable for its use in Bos indicus cattle breeds. Bos indicus cattle breeds are found to be genetically distinct from Bos taurus breeds. 1)SNP density in Bos indicus

It is approximately estimated that 1 snp in every 714bp in Bos taurus and 1 in 285 in Bos indicus (The Bovine hapmap consortium, 2009). As we know the whole genome size of bovine is about 2.8 billion base pairs, An average of 20.3- 29.3% of total SNPs in 777k chip and 35.52-45.5% of illumina snps on 50k chip were found to be monomorphic in Bos indicus breeds. An average of 19.7% of total SNPs in illumina 777k chip and 17.1 % of illumina snps on 50k chip were found to be monomorphic in bostaurus (HF) breeds. About 4% of the snps in the 75ki H was found to be monomorphic in Bos indicus and 33.5% in HF. here readers should not confuse by term monomorphic SNP and polymorphic SNP. SNP by deﬁnition is polymorphic but monomorphic SNPs show same genotype in all individuals which should be removed while analysing data because it is not informative. Only polymorphic SNPs that are heterozygotes while genotyping will be useful as it shows variation among individuals. So, Bos indicus chip like Geneseek 75Ki snp chip may be used in Bos indicus cattle breeds as it shows very less monomorphic SNPs. 2) Linkage Disequilibrium in Bos taurus and Bos indicus Linkage disequilibrium (LD) is a property of a contiguous stretch of genomic sequence that describes the degree to which an allele of one SNP is inherited or correlated with an allele of another SNP

within a population. LD is usually measured by |D| or correlation (r2) between two marker. But r2 parameter is most preferred as |D| tends to be overestimated in small samples. Many scientists have recommended correlation limit r2= 0.3 to have strong association with traits. Estimated LD between 2 snp is 14 kb with r2 = 0.2 in Bos indicus. LD between 2 snp is 29kb with r2 = 0.2 in Bos taurus. The low level of LD in Bos indicus indicates that indigenous cattle are in good effective population size, with more recombination between loci and lesser level of selection. Whereas in Bos taurus show high LD may be due to higher selection and use of very less males. The rate of LD decay is number of dependent on multiple factors, including the population size, the number of founding chromosomes in the population, and the number of generations for which the population has existed. Different cattle subpopulations have different degrees and patterns of LD. The most ancestral population has smaller regions of LD due to the accumulation of more recombination events in that group. 3) MAF Minor allele frequency is the frequency of the least common allele of a locus in the population. It ranges from 0.05-0.5. They are again classified into 3 categories (1) rare variant (0 - < 0.05), (2) intermediate variants (0.05-0.1), (3) common variants (0.1-0.5). Usually higher number of MAF is considered for involving a SNP in a chip. Challenges in developing SNP chips for indigenous cattle and buffalo In the present scenario the available SNP chips has been developed keeping in focus of the exotic cattle and buffalo populations. The research on the usage of existing SNP chips in indigenous livestock population revealed the limited efficiency to study accurate genetic diversity as well to estimate the GEBV of the population and thereby the genomic selection. Till date, Bos indicus SNP chips that include the Bos indicus specific candidate SNPs are not available for commercial usage. However, NDDB has DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

successfully designed INDUSCHIP based upon the SNPs drawn from that has got genotyped in Indigenous cattle populations. Moreover, the SNP density in case of Bos indicus in more than twice than the Bos taurus cattle breeds. In a whole genome sequencing study using Gir cattle, a total of 9990733 single nucleotide polymorphisms (SNPs) and 604 308 insertion/deletions (indels) were discovered. This study clearly indicates the higher level of SNP density in Indigenous cattle breeds. The linkage disequilibrium in case of exotic cattle is twice that of the Indigenous cattle, which clearly implies the need to include more number of SNPs in Indigenous SNP chip cattle breeds. The effective population size as well the minor allele frequency (MAF) is comparatively higher in Bos indicus which also insist the need for exclusive SNP chip for use in Indigenous cattle population. Buffalo speciﬁc SNP chip was developed recently using genomic information of several buffalo breeds (Italian Mediterranean, Murrah, Nili-Ravi,

Jaffarabadi, Kundhi, Aza-Kheli, Egyptiana and Swamp type from Philippines) that facilitated the identiﬁcation of millions of sequence variants in the buffalo genomes. Conclusion Widespread use of DNA markers has a major impact on the structure of the breeding programmes and a signiﬁcant impact on production systems. India has great opportunity to develop a high density (HD) SNP chips especially for indigenous cattle and buffalo apart from other livestock species. The huge investment can have better dividend for the country both in terms of increase in productivity as well as IP revenue. Such pragmatic approach with right objectivity and rationality is the need of the hour and is critically warranted. The low heritable traits like disease resistance

and reproductive traits can be improved with faster genetic gain than conventional breeding. The huge amount of investment incurred in genotyping a very large number of animals can be overcome by the higher genetic gain obtained through genomic selection. The new born male calf can be predicted for milk yielding breeding value. Breeding animals will be reared cheaply with minimum recording of phenotypes and pedigree. The SNP chips that's being planned for the Indigenous cattle and Buffalo population need to have uniform distribution of SNPs, high minor allele frequency and high call rate apart from considering the linkage disequilibrium.

Ravi kumar D¹, Vineeth Mr¹, Anshuman Kumar¹, Joel Devadasan¹, S K Niranjan² and Jayakumar Sivalingam², ¹ICAR-National Diary Research Institute, ²ICAR-National Bureau of Animal Genetic Resources, Karnal

DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

BENEFITS OF COW DESI GHEE

DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

TEA WASTE AS AN ANIMAL FEEDSTUFF

Tea / Coffee is a staple beverage in India as every Indian's day is incomplete without a cup of it. Tea makes a great alternative to coffee, as tea generally has lower caffeine content than coffee. Tea is predominantly harvested and processed in the states like in Assam, West Bengal, Tamil Nadu, Kerala, Karnataka, Tripura, Himachal Pradesh, Uttaranchal, Arunachal Pradesh, Manipur, Sikkim, Nagaland, Manipur, Meghalaya, Mizoram and Bihar. Currently there are more than 1600 registered tea manufactures, 2200 tea exporters and 5000 registered tea buyers all over India. Indian tea industry is the largest among the world with more than 13,000 tea gardens and has an annual production capacity of 1,300 million kg. In the course of production of tea there are large amount of byproducts being produced at various stages of processing. The waste includes discarded tea leaves, buds and tender stems of tea plants. The large portion of the by-product is going as a waste and a small fraction of it is utilized for caffeine extraction. There is an urgent need for proper disposal of tea waste as it is posing a serious environmental threat as a waste. Currently there is a huge gap of about 44% concentrate and 36% of green fodder is existing between the demand and supply of animal feed resources in the country. In this scenario utilization of tea waste as unconventional animal feedstuff is quite feasible affair. 22

In the course of processing of tea leaves processing at tea industries the fibre portion of the leaves are separated and discarded as a waste, which includes some tea leaves and dust. Decaffeinated Tea Waste (DCTW) is the waste available in the Caffeine factories after the extraction of caffeine from Factory Tea Waste. The industrial tea waste contains a fraction quantity of tannic acid (0.4–1.0% on a DM basis). Simple processing method like hot water soaking and washing makes the DCTW as good feedstuff for poultry, Pig and Fishes. Researches has shown improved immune response in finishing pigs and increased the egg laying capacity in hens which were fed with tea waste. Nutrient Profile of Tea waste The Tea leaf contains various chemical components like fibres, proteins, vitamins, minerals, amino acids, tannins and polyphenols. DCTW has moderate energy and protein source with 58% TDN and 11 % DCP. Tea waste on an average contain 15-20% crude protein and 1-4% amino acids, which includes theanine, aspartic acid, tyrosine, tryptophan, glycine, serine, valine, leucine and arginine. Also tea waste contains 5-7 % carbohydrates such as cellulose, pectin, glucose, fructose and sucrose and little quantity of fat in the form of linoleic and linolenic acids, sterols in the form of stigmasterol. DCTW are rich in vitamins B, C and E minerals (5 % DM) like Ca, Mg, Mn, Fe, Cu, Zn, Mo, Se, Na, P, Co, Sr, Ni, K, F and Al. Similar to black tea, Green Tea Grounds (GTG) contains 22-35 % of crude protein (CP), 2.3-7.1% ether extract (EE), 24-37% acid detergent fiber (ADF) and 31-45% neutral detergent fiber (NDF) on dry matter (DM) basis. GTG also contains tannin, caffeine, betacarotene and vitamin E. Similarly barley tea grounds (BTG) contains 12-19 % of CP, 2.2-3.4 % EE, 1525 % ADF and 27-35 % NDF on DM basis.

Proximate Composition of Tea waste

Moisture (%)

7.10

Crude protein (DM %)

19.99

Crude Fibre (DM %)

23.21

Ether extract (DM %)

2.18

Total Ash (DM %)

5.14

Nitrogen free extract (DM %)

49.48

Tea leaves waste contains xanthic bases l i ke c a f f e i n e a n d t h e o p h y l l i n e , p i g m e n t s l i ke c a r o t e n o i d s a n d chlorophyll, volatile compounds like aldehydes, alcohols, lactones, esters and hydrocarbons. These components have unique properties like lowering blood cholesterol level whereas the ﬂavanoids present in the tea waste exhibits anticarcinogenic, antimutagenic and cardioprotective effects as they act as antioxidant by scavenging free radicals. The green tea contains higher amount of catechin (ﬂavan-3-ol, a type of natural phenol) than black tea. Researches support replacement up to 20% of wheat bran in concentrate ration of calves. Also various feeding trials conducted proves that there is no adverse effect of inclusion of water treated tea waste in the ration up to 15% for Hampsire pigs. As far as the poultry is concerned the tolerance level of tea waste is 5 % inclusion in the feed. These parameters provide concrete evidence that tea waste is a superior unconventional feedstuff. Effective utilization of Tea waste as animal feed The palatability of tea waste is low as it contains tannin, therefore the tea waste must be mixed with the other palatable feedstuff for effective inclusion in the ration and to increase the intake of the same by the animals. The moisture content of the tea waste released from the industries is high hence they require proper drying before storage. Drying procedure incurs additional cost to the farmers therefore either they have to be DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

fed immediately or they have to be ensiled to preserve the moisture. The simple processing method to reduce the tannin content in the tea waste is to immerse in hot water (80Â°C) for 12 hours before feeding to animal. This simple method reduces the tannin content from 7 % to 3 % without affecting the crude protein content of the waste. Challenges The foremost problem associated with these waste is the transport cost from the processing site to the farmers. Also there is a great variability in quality of these tea wastes from one farm to another farm as the plantation and harvesting procedures varies. The tea waste is limited in the usage of poultry

feeds as they contain higher crude ďŹ ber (26%) and tannin (7-15%). Tannin impedes the absorption of nutrients as it is a poly phenolic secondar y metabolite which has a tendency to form complex with protein and carbohydrates. When animals are fed with feeds rich in tannin the feed intake, digestibility and rumen degradability reduces, also fecal N excretion increases. Feeds beyond 5% of tannic acid have deleterious effect on growth

and performance of chickens. Therefore the inclusion of tea waste should not exceed 10-15% as the TDN is low. Conclusion Tea waste is a major byproduct of different tea industries all over India. The huge amount of tea byproducts and wastes can be effectively utilized as livestock and poultry feedstuff so as to reduce the production cost of the feeds and to reduce the environmental impact of burning the waste or using it for landďŹ ll.

K. Rajkumar, A. Shanmuga Sundaram, C. Nithya and P. Tensingh Gnanaraj Livestock Farm Complex, Tamil Nadu Veterinary and Animal Sciences University, Chennai, Tamil Nadu

DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

Cows may seem like simple creatures - most of us have seen them grazing with seemingly not a care in the world. Well, there's more to these ruminants than meets the eye. Here are 20 facts you probably haven't heard about cows:

DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

Blue cheese canapes

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Ingredients l 2 cups (500 mL) cooked beets, coarsely chopped l Salt and freshly ground pepper l 6 slices of speck (smoked prosciutto) or prosciutto,

cut into strips l 30 lesley stowe's raincoast crisps® cranberry

hazelnut crackers l 30 baby arugula leaves l 2 oz (60 g) Blue cheese, coarsely crumbled

Directions Using a food processor, purée the beets; season with salt and pepper. Drain in a sieve for 5–10 minutes to remove excess water.Meanwhile, loosely roll up the speck strips to create rose shapes.Top each cracker with beet purée, a speck rose, an arugula leaf and Blue cheese. Serve immediately. Pixie Consulting Solutions Ltd. C/o OmAng Hotel, Namaste Chowk, Near Janta Petrol Pump, KARNAL - 132001 (Haryana) INDIA Email : dairy.pcsl@gmail.com | info@pixie.co.in Website : www.pixie.co.in

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DAIRY PLANNER | VOL. 16 | NO. 4 | APRIL 2019

NEWS/EVENT CALENDER DAIRY INDUSTRY AT RISK FROM HIGH IRRIGATION WATER PRICES farm gate milk price was likely to fall each year in real terms until then. With the pessimistic forecasts, dairy industry bodies will soon begin broad consultation with farmers, processors and service providers to develop its Australian Dairy Plan. About 20 meetings have been called for May and June to feed into the plan. Dairy Australia, Australian Dairy Farmers, the Australian Dairy Products Federation and the Gardiner Dairy Foundation want to identify key priorities to deliver “transformative and positive change for dairy” for the next ﬁve years and beyond.

he possibility of high irrigation water prices continuing into the next season may put a dampener on recovery of the dairy industry, according to Rabo bank. In the bank's latest Agribusiness Monthly report, senior dairy analyst Michael Harvey said further milk production losses in the Murray Dairy region could result if there was no respite in irrigation water prices. Dairy Australia ﬁgures show milk production in northern Victoria sliding monthly, with February's ﬁgures 26.4 per cent lower than the same month in 2017-18 and 17 per cent lower on a year-to-date basis. Milk production in nor thern Victoria has dragged down state production, which is now 7.8 per cent lower than last season. With both NSW and Queensland milk production 10.1 per cent lower than last s e a s o n , ye a r - to - d a te n a t i o n a l m i l k

production is 6.4 per cent lower than last season. Rabo Research is forecasting milk production to ﬁnish the season down 8 per cent, meaning that milk production numbers will get worse before they get better,” Mr. Harvey said. The rapid decline in milk production leaves Australian dairy supply chains short of milk solids.”The outlook in the immediate future is not good. At its annual Outlook Conference in Canberra last month, the Australian Bureau of Agricultural and Resource Economics and Sciences forecast dairy cow numbers to c o n t i n u e f a l l i n g t h ro u g h t o 2 0 2 1 22.ABARES said yield increases through natural productivity gains were unlikely to offset falling cow numbers, with national milk production to fall to 8.6 billion liters by 2021-22 and remain below nine billion liters at least until 2023-24.It said the Australian

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