Veterinary College, Bengaluru Newsletter Date : 31st December 2013
Volume No: 2 Issue :12
Dr. B. N. Nagaraja, Dr. A. S. Patil, Dr. Ramesh Rathod and Dr L. Ranganath Department of Veterinary Surgery & Radiology, Veterinary College Hebbal, Bangalore (settihallynag@rediffmail.com)
Fractures of long bones are encountered on regular basis in large animal practice and managing them poses a challenge to field veterinarian. Dealing with long bone fracture will always remain a challenge for the clinician and cost of treatment has to be clarified with the owner before starting it as we should ensure the animal worth the investment to make relatively to the prognosis. The incidence has increased in recent years due to increased vehicular traffic. Injury such as accidents, fall, fighting, kicks, forceful muscle contractions, mounting, or dismounting Fractures in racing bullocks happen due to low mineral density and the quality of bones which depend on many factors such as inadequate intake of calcium and Vitamin D3 and lowered levels of physical activity, low plane of nutrition etc. Cattle are considered great orthopedic patients. Most of them tolerate orthopedic devices to stabilize the fracture site and have a prodigious potential of bone healing. As they lay down several hours a day, they protect the fracture site and are less prone to suffer of contra-lateral limb diseases. Yet no suitable treatment is available for fracture in large animals. Fractures in large animals are difficult to treat due to their heavy body weight, temperament, poor muscle covering on certain bones, angular placement and conical shape of hind limb. Prognosis is generally poor in cases which remain recumbent for more than three days. However for all cases, dealing with an open fracture in cattle will lower the prognosis from overall good to guarded whatever is the degree of contamination. Evaluation of fracture should be done and at least two orthogonal radiographs have to be taken to determine the type of fracture and the degree of comminution and displacement of the bone segments.
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In addition to that, open fractures are classified following the Gustilo and Anderson classification. Open fracture type I refers to a wound less than 1 cm associated with minimal soft tissue damage and a relatively clean wound bed. Open fracture type II follows the type I characteristics with the exception of the wound’s length (>1 cm) and a moderate soft tissue damage. The type III is associated with extensive damage to the soft tissue compromising the blood supply to the distal aspect of the leg, exposed bones and massive contamination. Type III open fractures are usually due to a high velocity trauma or a severe crushing component and therefore are severely comminuted fractures. Long bone fracture management in cattle varies from stall rest, external coaptation, external fixation to open reduction and internal fixation depending on the type of fracture, the bone involved and is essentially driven by the economic limitation. Splint and plaster casts are the two of the oldest methods of fracture management employed. Plaster of Paris (POP) cast alone or Thomas splint application will not yield adequate fracture reduction and immobilization in large animals. Moreover, it lacks mechanical strength, cannot be used as a sole device of fracture treatment and becomes wet in case of compound fracture. Fenestrated cast are difficult to manage adequately and should be discouraged: they offer limited access to the wound and the hole in the cast acts as a weak point in the external coaptation system weakening the fracture stabilization. Thomas splint and plaster cast combination has given fairly good results for fractures of the radius, ulna and tibia in cattle (Fig. 1 and 2).
Fig. 1. Modified Thomas splint for For Radius and Ulna Fracture management
Fig.2. Combination of Modified Thomas splint and plaster of paris cast for Tibia Fracture Management
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For POP cast application joins on either side of the fractured bone should be immobilized. The part of the limb below the bandage should be carefully and firmly wrapped with an ordinary cotton bandage all the way from the plaster bandage down to the hoof. This last bandage will tend to prevent swelling. When plaster of Paris bandages are applied to a compound fracture the injured part may be previously dressed with a small, thick pad of cotton immediately over the wound. Then a window is made for regular dressing. The ends of the pop bandage should be carefully watched to see that the skin does not become chafed, particularly at the lower end. Either cast or modified Schroeder-Thomas splint-cast combinations may interfere with ambulation and result in prolonged recumbency, decubital ulcers, joint stiffness, muscle atrophy, and delayed fracture union. External skeletal fixation (ESF) refers to stabilization of fractured fragments using transcortical pins connected externally with any solid coonecting frame / bars. ESF has been tried with transcortically passed pins reinforced externally with several methods such as wooden frame, iron frame, plaster cast, PVC pipe, epoxy putty or poly methyl methacrylate frame on either side. It allows diverse designs for treating comminuted fractures, permits wound dressing of limb in case of compound fracture, permits local blood flow to the fracture site and provides early return to function of fractured limb (Fig. 3 and 4).
Fig:3. External skeletal fixation Type II for
Fig:4. External skeletal fixation Type II for
stabilization of tibial fracture in H.F cow
stabilization of metacrpal fracture in khillar
The ESF construct should be maintained for at least 10-12 weeks. However, ESF is also associated with several disadvantages such as sub-optimal reductions of fracture, poor anatomic alignment, absence of inter fragmentary compression and less rigid fixation when compared to bone plating. Recently more and more designs of external skeletal fixators such as dynamic axial fixators and circular fixators have been fabricated.
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Volume No : 2 Issue : 12 09 11
They have not been tested largely in heavier cows or bullocks. In the absence of such devices, it can be concluded that trans-fixation pinning along with wooden frame, plaster cast or PVC cast may be employed for selected cases of fracture as it provides a better fixation than plaster cast alone, permits wound management at the fracture site and can be undertaken at field level if learnt and applied correctly to manage fractures of large animals. Loosening of one or more pins and wetting of plaster due to pus discharge are the common complications. Animals less than 250 kg may be treated with internal fixation. Animals heavier than 250 kg have such great muscle mass that makes internal fixation is difficult. Also, the bone implants are not of sufficient strength without double plating. However, bone plating is not feasible for heavier animals, in compound fractures and is not economical. The technique is not easy to practice at field level. Plating long bones is not recommended in neonatal calves because the cortical bone is too soft to retain screws. Intramedullary pins have been used to repair humeral fractures; the pins are used to fill as much of the medullary cavity as possible. The smaller and younger the animal, the greater the chances of success. The fracture must be close to the mid-diaphysis and not comminuted to allow repair with intramedullary pins. Stainless steel wire can be used as cerclage in combination with intermedullary pins or lag screws if the fracture has definite oblique spiral components. Considering the typical environnement of cattle, any penetrating wound should be considered infected. A mixed bacterial flora can usually be cultured from a swab sample from the wound bed. Common bacteria isolated are coliforms, Arcanobater pyogenes, Staphylococcus spp and Streptococcus spp. The control the infection is the main target to reach the ultimate goal of fracture healing. Surgical debridement should be considered as the most effective way to reduce the bacterial load of the wound. The wound is copiously lavaged with a poly-ionic solution using a simple lavage system to add a mechanical action to the sharp debridement. Antibiotherapy should be started immediately. The initial broad spectrum therapy should be reassessed depending on the bacterial culture and sensitivity results of the sample obtained at the time of the surgery. Local antibiotic delivery is achieved with slow release implants in situ or local injections. Antibiotic impregnated implants (collagen sponges, polymethylmethacrylate) can be inserted at the time of the surgery if the wound is closed and first intent healing process is targeted. Increase of the antibiotic concentration in the wound bed and bones can also be performed using regional IV perfusion distal to a tourniquet or intraosseous perfusion. Unlike western countries where large animals with fractures are slaughtered, in India owing to economic and religious reasons livestock owners force for treatment till hope. However, the decision between treatment and euthanasia depends on the economic or genetic value of the animal, the cost of the treatment and the prognosis associated with the particular fracture.
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Volume No : 2 Issue : 12 09 11
Dr. Madhukar* and Dr. H. A. Upendra# *
#
Assistant Professor, The Director, Institute of Wildlife Veterinary Research, KVAFSU, Doddaluvara, Kodagu – 571232. (madhukar262@gmail.com)
India is home for nine species of vultures. Three of these species, White-backed Vulture, Slender billed Vulture and Long billed Vulture declined drastically with the genus Gyps population falling by 97% by 2005. Looking at this rapid decline, International Union for Conservation of Nature (IUCN) declared the three vulture species Critically Endangered, which is the highest category of endangerment. Hence, it was clear that, if no immediate action was taken, these natural scavengers would soon get extinct. Repeated scientific investigation showed that diclofenac residues in livestock carcasses were leading to numerous effects in vultures and large scale deaths. Hence, IUCN passed a motion in 2004 and “called upon Gyps vulture Range countries to begin action to prevent all uses of diclofenac in veterinary applications that allow diclofenac to be present in carcasses of domestic livestock available as food for vultures; establishment of IUCN South Asian Task Force under the auspices of the IUCN; Range countries to develop and implement national vulture recovery plans, including conservation breeding and release.” Vultures are primary removers of carrion in India which keeps environment clean. Also, Parsi community disposes their dead ones by allowing vultures to feed on the dead bodies. Hence, vultures have enormous ecological, social and cultural importance. Decline in vulture population is affecting the equilibrium of other scavengers, increasing the number of putrefying carcasses. Population of feral dogs and rats is believed to be increasing due to vulture decline, which is estimated to be costing over Rs. 8000 crores every year in the form of rabies, other diseases and effects. Vultures do not cause any diseases to humans or other animals and clean the carcasses completely, whereas other scavengers are not so efficient and have tendency to transmit or produce deadly diseases. Parsi community is depressed as it has become difficult for them to dispose the dead ones as per their rituals. Scientists from Bombay Natural History Museum, IVRI and many other institutes were successful in rapidly identifying acute visceral gout and renal failure as cause of vulture population crash. Extensive investigation after this starting point showed that this was caused by diclofenac, which was used in veterinary patients since 1980’s, to which vultures were exposed by consumption of livestock carcass treated just before death with diclofenac. This hypothesis was reconfirmed by laboratory studies. Very unfortunately, very low levels of diclofenac in carcass are enough to cause massive decline in vultures, as diclofenac is highly toxic to vultures, which is complimented by group feeding habit of vultures.
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Volume No : 2 Issue : 12 09 11
After identification of diclofenac as a major culprit, a large number of workshops and meetings were conducted by policy makers, scientists and conservationists to find out possible steps to reverse the vulture decline. Two decisions were taken during these meetings, one to initiate large scale captive breeding of vultures and second to ban and replace veterinary use of diclofenac with safer alternatives like meloxicam. Consequent to this, Animal Husbandry and Veterinary Services departments of all the sate were directed to stop procuring diclofenac. Further, all the veterinarians were instructed to refrain completely from diclofenac use.
Figure: A Slender-billed Vulture killed by Diclofenac poisoning. Photo credit – Devojit Das / BNHS.
After 5 years of diclofenac ban, follow-up studies done in 2011-12 showed that diclofenac contaminated carcasses reduced by more than half but not hundred per cent. There is a continued presence of diclofenac in many carcasses which indicates that the problem has not yet been overcome. As very low number of contaminated carcasses is enough to kill vulture populations, the continued use of diclofenac is still killing many birds annually. Surveys of pharmacy shops in India showed that veterinary labelled diclofenac was rarely offered for sale for use on livestock after the 2006 ban, however very unfortunately, human formulations of the drug were being sold widely for veterinary use. Hence, it is time for the veterinarians to introspect in this matter and make sure that diclofenac is not used by them or their subordinates in their jurisdiction. This is the only way large scale breeding programmes can be complemented to recover the vultures from the tip of extinction. It would be a matter of great pride for all the veterinarians of India to act at their own level and see that diclofenac use is completely phased out, which will ensure reversal of the vulture decline. This contribution to the conservation of vultures and the ecological balance at large will itself be an honour to the efforts of a veterinarian.
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Volume No : 2 Issue : 12 09 11
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Pashubandha 2013
Volume No : 2 Issue : 12 09 11
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Pashubandha 2013
Volume No : 2 Issue : 12 09 11
ಆ?ೆK/ೋ•ಾOೆ}€ ಗಳH ತುಂRಾ
ಷ:ಾa„ಾ7ದುh, ಇದರ
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eಾನು ಾರುಗಳH 5ಂ ಾಗ ಇ•bೋ:ೊ@ೕ‘+ಡ!ಳ
ಷ ಾ ೆಯನ2 :ಾಣಬಹುದು.
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ಮುಂXಾದವJಗಳH. ಇವJಗಳನು2 .ೈತ ಸಮು ಾಯದ •ತ+ರು ಇ=ಗfಂದ ತಮi ದವಸ ,ಾನ ಗಳನು2 / Xೋಟ?ಾa:ಾ RೇSೆಗಳನು2 ಸಂರ–ಸಲು rನಗಳ ನಂತರ
ಉಪZೕ7ಸುXಾ.ೆ.
ಾ•ೆKaV ಗಂ‘?ೆ Oೇaದ
ಇ= /ಾಶಕಗSಾ7ದh=@, Oೇ Fದ 3-4
, ಮೂಗು, Rಾp ಮತು ಗುದ ಾ‰ರಗfಂದ ರಕOಾ+ವ ಾಗುತ ೆ,
ಗಂyೕರ Fo5ಯನು2 ತಲಪJತ ೆ. ಇ=Tಾ ಾಣಗಳH ೇಹದ=@
ೇಹದ
ಉಷjXೆ ಕE<„ಾ7,
ಟ•V- :ೆ ಯ Oಾ[ಾನ :ೆಲಸ:ೆ^ ಅE{ಯುಂಟು[ಾE,
ರಕ ೆಪJ•ಗಟುnವJದನು2 ತbೆಯುತ ೆ, ಾ?ಾ7 ೇಹದ=@ ರಕ ಪaಚಲ/ೆಯ ವ Xಾ ಸಯುಂgಾ7, 5ವ+ ರಕOಾ+ವವಗುತ ೆ. eಾನು ಾರುಗಳH ಇ=TಾUಾಣಗಳನು2 5ಂದ 2-3 rನಗಳ ನಂತರ
ಷRಾ ೆಯ ಲqಣಗಳನು2 Xೊaಸುತ ೆ.
ದ=?ೆ
<ೕವJ 5ನು2ದನು2 A=@ಸುತ ೆ, :ೆಮುi, ೇಹದ ಉUಾjಂಶ ಕE<„ಾಗುವJದು,ರಕ • +ತ ಮೂತ+,xಂRಾಗದ ವೃಷ™ವನು2 ತುಂRಾ ಗMn„ಾ7ಸುವJದು, ಇದh ^ದhಂXೆ Oಾವನ2ಪJ•ವJದು. 3. Gಾ+ಾಯIಕ 1ೊಬJರಗಳ& (ಫM2 ೈಜರುಗಳ&): ?ೊಬŽರಗಳನು2
`ೕಲಗfಂದ
ೊರ7ಟn
ಯೂ4Oಾ, 7ಾ+ೆ8ೕಟ<ಳ&, P.ಎ.B, ಮುಂXಾದ .ಾOಾಯAಕ
ಸಂದಭKದ=@
ಷRಾ,ೆಯುಂgಾಗಯತ ೆ. .ಾOಾಯAಕ ?ೊಬŽರಗfಂದ
eಾನು ಾರುಗಳH
ಉಂgಾಗುವ
ಅಕFiಕ ಾ7
ಷRಾ,ೆಯ=@ eಾನು ಾರುಗಳH
5ಂ ಾಗ ಪaೕತ
eೋಲು@ ಸುaಸುವJದು, ೊgೆn ಉಬŽರ, ಉr:ೊಂಡ ಕಣುj ಗುbೆ{ಗಳH, ೊgೆn /ೋ Aಂದ ಮಲಗುವJದು, ಏಳHವJದು, ರಕ Rೇr ಇXಾ r ಲqಣಗಳನು2 /ೋಡಬಹುದು..
4. ಕRೆ!ಾಶಕಗಳ& : ಆಸKAಕ…š, 2,4-E, 3,4,5-M, ಡ„ಾ:ಾ‰€, Tಾ .ಾ:ಾ‰€, OೋEಯಂ :ೊ@ೕ.ೇ€, ಅgಾsV, FನsKV, ಮುಂXಾದವJಗಳH.bೈ/ೈgೊ+ೕ ಕSೆ/ಾಶಕಗಳ
ಷRಾ,ೆpಂದ ಬಳಲುವ eಾನು ಾರುಗಳ=@ ಹಳr- ಹFರು ಬಣj:ೆ^
5ರು7ದ ಮೂತ+, :ಾ[ಾdೆ .ೋಗ, ಕರುಳH Rೇ/ೆ, ಚಮKಸುಕು^ಗಟುnವ ಲqಣಗಳನು2 :ಾಣಬಹುದು.
Pashubandha 2013
Volume No : 2 Issue : 12 09 11
5. TDೕಂಧ !ಾಶಕಗಳ&: ಕ/ಾKಟಕದ ೇಂ?ಾ
ಧ sdೆ@ಗಳ=@ ಇ5ೕuೆ?ೆ ಹಲ ಾರು eಾನು ಾರುಗಳH
ೊಟುn ಇವJಗಳನು2 5ಂದು
ರOಾಯAಕಗಳH,
ಷRಾ,ೆ?ೊಳ?ಾದ ಹಲ ಾರು ದೃUಾ™ಂತಗಳನು2 ಗಮAಸdಾ7 ೆ. Tಾದರಸಯುಕ
:ೊ@ೕ.ೋœ/ಾಲ!ಳH(Tೆಂgಾ:ೊ@ೕ~
œ/ಾ…
/ೈgೊ+ೕœ/ಾ…, ಥ„ಾ.ಾ“, ಸಲ}~ ಇXಾ r. ಅ5„ಾ7 eೋಲು@ ಸುaಸುವJದು, ಪ ೇ ಪ ೇ ಮೂತ+ ವ Xಾ ಸ
=ೕಂಧ+ RಾŒತ ಬತದ ಹುಲು@, eೋಳ,
ಮತು
=ೕಂಧ+/ಾಶಕಗಳ
OೋEಯಂ
Tೆಂgಾ:ೊ@ೕœK/ೇ€),
ಷRಾ,ೆ?ೆ ಒSಾ?ಾಗುವ ಕುa/<ೕ:ೆಗಳH
ಸಜK/ೆ, ಾಸ/ೆpಂದ ಕೂEದ ೇr, ಕ•jನ TಾTೆಯ ?ಾತ+ದ=@
ಾಗೂ ಕ•jೕರು ಸುaಸುವJದು, Oಾಯು2 OೆSೆತ
ಾಗೂ <ೖನಡು7ಸುವJದು, ಉ ೆ+ೕಕ?ೊಳHI :ೆ ಅಥ ಾ Rೆ`4
yೕಳHವJದು ಮತು ಉF.ಾಟದ Xೊಂದ.ೆಯಂತಹ ಲqಣಗಳನು2 :ಾಣಬಹುದು.
ಷWಾXೆಗಳ ತ ೆಗಟು* 1.
ೕಟ/ಾಶಕಗಳನು2
ೆ1ೆ ಅನುಸ4ಸZೇ ಾದ ಮುಂ]ಾಗ ತ ಕ ಮಗಳ&:
eಾನು ಾರುಗf?ೆ FಗದಂXೆ ಸೂಕ eಾಗದ=@E, ಆ ಾರ/ <ೕ /ೊಂದ?ೆ „ಾವJ ೇ ತರದ
ೕಟ/ಾಶಕಗಳನು2 ೇಖaಸಡRೇE. 2.
ೕಟ/ಾಶಕಗಳ Fಂಪರtೆ?ೆ ಉಪZೕ7Fದ TಾXೆ+, ಬ:ೇಟು ಮುಂXಾದವJಗಳನು2 ಸa„ಾ7 ಶು`?ೊfಸ ೆ eಾನು ಾರುಗf?ೆ ಢAೕರು ಅಥ ಾ ಆ ಾರವನು2 AೕಡRೇE.
3. Ÿಾ=„ಾದ ೕಟ/ಾಶಕದ FೕOೆ, 4. 5.
ಟnಣಗಳನು2 ೊಲದ=@ ಗುಂE Xೆಗುದು ಹೂfa.
ೕಡ ಅಥ ಾ ಅ5„ಾದ Xೇ ಾಂಶ ದh=@ ೕಟ/ಾಶಕ ಬಳಸುವJದನು2 ಮುಂದೂE . ೕಟ/ಾಶಕಗಳನು2 Aಗrತ ಪ+[ಾಣದdೆ@ ಉಪZೕ7F. <ೖ<ೕdೆ ?ಾಯ/ಹುಣುjಗSಾ7ದh=@
ೕಟ/ಾಶಕಗಳನು2
ಬಳಸRೇE. 6.
ೕಟ/ಾಶಕ ಉಪZೕ7Fದ <ೕdೆ eಾನು ಾರುಗಳH <ೖ /ೆಕ^rರುವಂXೆ /ೋE:ೊfI.
7.
ೕಟ/ಾಶಕ ೆ`4ನ ಪ+[ಾಣದ=@ ಚಮKದ ಸಂಪಕK:ೆ^ ಬಂದ.ೆ Oಾಬೂನು AೕaAಂದ XೊSೆದು, ?ಾfಯ=@ ಒbಾಡಲು yE.
8. ಕೃ*?ೆ ಬಳಸುವ
ೕಟ/ಾಶಕಗಳನು2 ಕುa/ <ೕ:ೆಗಳ ಪ.ೋಪsೕ ಗಳ Aಯಂತ+ಣ:ೆ^ ಬಳಸRೇE, ಏ:ೆಂದ.ೆ ಕೃ*?ೆ
ಬಳಸುವ ೕಟ/ಾಶಕಗಳH ಕುa/ <ೕ:ೆಗಳ=@ ವ 5aಕ ಪatಾಮವನು2ಂಟು[ಾಡುತ ೆ.
Pashubandha 2013
Volume No : 2 Issue : 12 09 11
Dr. Suguna Rao Department of pathology, Veterinary College, KVAFSU, Bangalore. (sugunabg@yahoo.com) The clinical and pathological features of a number of diseases of animals are so similar and overlapping that an accurate diagnosis could be made only by the employment of a number of laboratory diagnostic techniques. The value of laboratory diagnostic tests depends upon the proper selection, collection, preparation, packing and transport of specimens. Diagnostic laboratories require the submission of appropriate samples in good condition for testing. Hence the samples should be carefully packaged, labeled, and transported to the laboratory by the fastest practicable method with an appropriate temperature control. Considerable skill and care are required to decide on the correct samples to be sent to the laboratory. The samples collected should be representative of the condition being investigated and the lesions observed. Also the stage of the disease and lesion development should be considered, as well as the type of test(s) that will be performed while collecting the samples. Frequently, a combination of blood samples for serology and tissues from dead animals will be required for microbiological culture and pathological examination. •
Specimens for laboratory tests should be collected as early as possible after death of the animal as materials showing decomposition are usually unsuitable for diagnostic purpose.
•
Animal health personnel should be trained in the correct procedures for post-mortem examination of the species of animals with which they work.
•
The equipment required will depend on the size and species of animal, but a knife, saw and cleaver will be required, and also scalpel, forceps and scissors, including scissors with a rounded tip on one blade, for opening intestines.
•
A plentiful supply of containers and tubes of transport media appropriate to the nature of the sample required should be available, along with labels and report forms.
•
Containers should be fully labelled with the date, tissue and animal identification.
•
Special media may be required for transport of samples from the field.
•
The person conducting the post-mortem examination should have sufficient knowledge of anatomy and pathology to select the most promising organs and lesions for sampling.
•
Each piece of tissue should be placed in a fully labelled separate plastic bag or sterile screw-capped jar. Swabs should always be submitted in appropriate transport media.
Pashubandha 2013
Volume No : 2 Issue : 12 09 11
The specimens to the laboratory should be submitted along with all the information regarding the material and should include, • A full description of the animal • Date and time of death, and date and time of collection of materials • Owner’s name and address • Clinical signs and autopsy findings • Nature of the specimen • Disease suspected • Type of examination required. Many varieties of containers are available commercially for collection and storage of samples which include vials, test tubes, bottles, disposable variable sized containers, milk sample bags, catheters, sterile syringes, swabs etc. At post mortem samples may be collected for variety of laboratory examination such as bacteriological, virological, immunological, toxicological, cytological, parasitological, histopathological, biochemical, for detection of proteins or genome nucleic acids etc. for which the type of sample and methods of collection and preservation differs. Samples for bacteriological examination •
Materials for bacteriological examination should be collected under aseptic precautions to avoid contamination as much as possible. • Suitable sterile instruments should be available in sufficient numbers such as scalpels, scissors, forceps, knives etc. Care should be taken not to contaminate samples with intestinal contents. Disinfectants should not be used on or near the tissues to be sampled for bacterial or viral isolation. • Sufficient number of sterilized containers of different types depending upon the type of material to be collected should be available for collection and storage such as petri dishes, test tube, vials, wide mouthed bottles, swabs etc. • Samples to be collected for bacteriological examination include blood from the heart, fluids from the cavities, joints, urine, faces, CSF, exudates, intestinal and stomach contents and tissues from organs. •
Blood from the heart should be collected after singeing the unopened right ventricle with a red hot spatula by aspiration using a sterile syringe and needle.
• •
• •
Fluids from the cavities, urine from urinary bladder, CSF, synovial fluids also should be collected using sterile syringe and needles. For tissues from organs, prior to collection the surface of the organ should be seared with a hot spatula and the singed area should be punctured with sterile instrument and deep tissue should be scooped. Exudates like pus could be collected using sterile swabs. Intestinal and stomach contents could be collected by tying the ends of intestinal loop or stomach and cutting the tied ends and transferring into a wide mouthed bottle.
Pashubandha 2013
Volume No : 2 Issue : 12 09 11
The samples for bacteriological culturing could be directly placed in to a sterile container containing transport medium. Most bacteria could be isolated from the Tryptose broth base media. After collection the sample containers should be held under refrigeration until received in the laboratory which could be achieved by the use of ice packs or dry ice.
Samples for Virological examination •
Specimens for virological examination should be as free as possible from bacterial contamination. Utmost care should be adapted in selection of specimens appropriate for the disease. Ex: brain tissue for rabies, CD; skin and lungs for sheep pox
•
The tissue specimens should be placed in sterile, wide mouthed tightly closed bottles and shipped frozen with dry ice or in 5 – 10 volumes of sterile 50% glycerol saline if refrigeration is not possible.
• •
Tissues also could be collected in culture medium like Hank’s balanced salt medium. Heart blood and fluid specimens also could be collected appropriate to the viral infection under aseptic conditions and transported in sterile containers under refrigeration.
Materials for Toxicological examination Death due to poisoning in animals is usually due to consumption of poisonous plants or vegetation contaminated with insecticides accidentally or sometimes due to malicious intention. •
•
Materials to be collected in suspected poisoning cases include, •
Stomach and its contents
•
Small intestine (upper part about 25 – 30 cms) and its contents
•
Liver (about half kg) – large pieces
•
Kidney (half of each)
•
Blood (about 50 – 100ml) minimum of 10 ml should be collected.
•
Urine – 100 ml
As most of the poisons are taken orally, stomach and intestinal contents along with respective organs should be collected. As liver is the organ of detoxification and kidneys are the excretory organs, liver, kidneys and urine should be collected.
•
The tissues and contents should be collected in large containers (one litre capacity) wide mouthed, clear transparent with glass stoppers as rubber stoppers may extract some poisons such as chloroform and phenols. Polythene bags and containers also could be used. For volatile poisons, nylon bags should be used as they are not permeable to such substances.
Pashubandha 2013
Volume No : 2 Issue : 12 09 11
Preservatives that could be used for dispatch of materials for toxicological examination include •
Saturated sodium chloride solution
•
Rectified spirit ( except in alcohol, acetic acid, phenol, phosphorus and paraldehyde poisoning)
•
Sodium or potassium fluoride (10 mg/ml) for blood.
Dispatch of materials •
Stomach and its contents, intestine and its contents can be collected in one bottle/ container, liver and kidney in another container, urine and blood separately in different containers.
•
The quantity of preservative should be equal to the volume of the tissue or contents collected
•
Containers/bottles should be filled only up to 2/3rd of its capacity to avoid bursting of bottle if gases of decomposition are formed.
•
The bottles should be well stoppered, covered with a piece of cloth, tied by a tape/string and ends should be sealed in Veterolegal cases.
•
The bottles should be labeled giving the details regarding animal, organs collected, date and time of collection, type of poison suspected etc.
•
A sample of preservative used (100 ml of rectified spirit, 25 g of NaCl) should be sent separately to exclude the possibility of any poison being present as a contaminant.
•
A copy of requisition letter should be sent.
Materials for Parasitological examination The type of samples for parasitological examination depends upon the type of parasitic infection. In hemoprotozoan infections, microfilariasis and others blood smears should be made. •
Ectoparasites could be obtained by the keen observation of hair coat and by brushing, combing and shaking the fur or plucking the parasites. Ectoparasites like ticks, lice, fleas should be collected in vials containing 70% ethyl alcohol.
•
In mange, mites could be collected by deep skin scrapings in 5-10% sodium or potassium hydroxide for examination and could be preserved in 5-10% formalin.
•
Endoparasites like trematodes, cestodes and nematodes should be collected in 5-10% formalin.
•
In coccidiosis, the faecal sample should be collected in 2% potassium dichromate.
Pashubandha 2013
Volume No : 2 Issue : 12 09 11
Materials for Immunological examination •
Serum for lab examination could be obtained by collecting the plasma clots, unclotted blood or clotted blood from the right ventricle of heart and should be kept undisturbed for 30 minutes for clot formation and then centrifuge at 2000 G for 20 min. The serum obtained should be collected in vials and then refrigerated or frozen (-20 or -80) and transported to a laboratory.
•
Serum samples could be used for biochemical and serological tests.
Materials for Molecular tests •
For polymerase chain reaction the tissue samples from the organs appropriate to the infection should be collected in vials and frozen at -80.
Materials for Cytological examination •
For cytology impression smears or scraping smears from the affected organ could be collected. A small piece of tissue from the affected organ should be held with a forceps, the cut surface should be blotted and then gently pressed on a clean slide to obtain the impression of the issue.
•
For scraping smears the surface of the affected organ should be blotted and then gently scarped using a scalpel or knife and the scraped tissue is used to make smear on a clean slide.
Materials for Histopathology Samples for histopathology should be collected from all major organs and from all abnormal areas. All tissue samples should be placed in a common container containing 10% Neutral Buffered Formalin. 10% buffered neutral formalin (10% BNF) Probably the best routine fixative for histopathology, though penetrates tissue slowly (~5 mm/24 hours). Commercial Formaldehyde (37-40%)
100 ml
Distilled water
900 ml
sodium phosphate monobasic
4.0 g
sodium phosphate dibasic (anhydrous)
6.5 g (pH should be 7.2 ± 0.5)
Fix tissue slices 24 - 48 hours at room temperature. The tissues should not be thicker than 0.5 cm so that they can be fixed properly and should include the lesion and adjacent normal area. The samples should be handled carefully by grasping at the edges. Do not scrape surface of tissue or compress with forceps. The tissues should be placed in 10 times the volume of formalin as the volume of the tissue. If a tissue needs special labeling like specific lymph node, collect in a different container. At post mortem care should be taken not to lose any tissue/sample appropriate for definite diagnosis due to negligence as once the body is disposed the possibility of making accurate diagnosis of cause of death is also lost.
Pashubandha 2013
Volume No : 2 Issue : 12 09 11
Fodder Museum at Veterinary College, Hebbal, Bangalore
monthly e-Bulletin Published and circulated by Veterinary College, Hebbal Bengaluru Editor:
Associate Editior:
Dean, Veterinary College, Hebbal, Bengaluru Dr.S.Yathiraj (Ex-Officio)
Head,Dept of Vety & Animal Husbandry Extension Education Dr.K.Satyanarayana (Ex-Officio)
Contact : Dept of Veterinary and Animal Husbandry Extension Education Veterinary College, Hebbal Bangalore email: pashubandhavch@gmail.com
â&#x20AC;˘
2012 Pashubandha 2013
PELVIC
07 09 Volume No : 21 Issue : 11