Journal of Indian Veterinary Association December 2012, 10 (3) by magnus paul

Vol. 10 Issue 3 December 2012

ISSN 0975-5195

JIVA Journal of Indian Veterinary Association Kerala www.jivaonline.net

Editorial Board Chairman

Dr. K. R. Arun Kumar President, Indian Veterinary Association, Kerala

Editorial Advisor

Dr. K. Vijayakumar, M.V.Sc.,Ph.D Joint Commissioner of Animal Husbandry Gol, New Delhi

Chief Editor

Dr. A.P. Usha, M.V.Sc., Ph.D Editor

Dr. Laiju M. Philip, M.V.Sc. Associate Editors

Dr. Hiron M Harshan, M.V.Sc.,Ph.D Dr. M. Ranjith Mohan, M.V.Sc. Members

Dr. P.M. Hari Narayanan, M.V.Sc. Dr. M.G. Sajesh, M.V.Sc. Dr. Bindya Liz Abraham, M.V.Sc.,Ph.D Jivaonline.net Editor

Dr. K. Magnus Paul, M.V.Sc. Convener

Dr. C. Sreekumar General Secretary Indian Veterinary Association, Kerala

INDEXED IN CAB INTERNATIONAL, EBSCO, HINARI AND INDEX COPERNICUS INTERNATIONAL

Guidelines to Authors

About us Journal of Indian Veterinary Association, Kerala (JIVA), the official organ of Indian Veterinary Association, Kerala is a scientific periodical with international status (ISSN-0975-5195) which bring about the latest updates in Veterinary Medicine and Animal Husbandry Practices. The journal covers almost all topics of Dairying and Animal Husbandry besides special emphasis on Companion Animal Medicine and Surgery, Zoo and Wildlife Medicine, Meat and Feed industry, Diagnostics and Bioinformatics. The Journal JIVA is in wide circulation among all stakeholders of Veterinary Medicine and Animal Husbandry sector including the Veterinary Professionals working with Animal Husbandry Department, Kerala Veterinary and Animal Sciences University, Kerala Agricultural University, Kerala Livestock Development Board, Milma, Techno Park, Veterinary and Medical Research Institutes of the country. The Online edition is available at www.jivaonline.net JIVA is indexed in CAB International full text, HINARI (World Health Organization), EBSCO (World's Foremost Premium Research Database Service), and INDEX COPERNICUS INTERNATIONAL.

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CONTENTS RESEARCH ARTICLES 1.

5 - 51

BETA CASEIN A1A2 POLYMORPHISM AND MILK YIELD IN VECHUR, KASARGODE DWARF AND CROSSBRED CATTLE 5

E. M. Muhammed and M. Stephen

A HISTOLOGICAL STUDY OF THE STRUCTURAL CHANGES IN THE PANCREAS OF DIABETIC RATS 10

B. Dhanush Krishna and Suguna Rao

THERAPEUTIC POTENTIAL OF AFANIL速 AGAINST BLOAT AND FOR EARLY RESTORATION OF RUMEN FUNCTION IN BOVINES 15

S. U. Digraskar, V. D. Muley, K. Ravikanth, M. Dandale and S. Maini

OPTIMIZATION OF PCR THROUGH MANIPULATION OF CYCLE TIMES AND INCLUSION OF FORMAMIDE 19

S. Pramod, A. P. Usha, T. Venkatachalapathy and K. C. Raghavan

MILK FEEDING STRATEGIES FOR IMPROVING GROWTH PERFORMANCE IN CROSSBRED DAIRY CALVES 24

A. J. Flamy and Joseph Mathew

MORPHOGENESIS OF MEDULLA OBLONGATA IN GOAT FOETUSES 28

K. M. Lucy, K. R. Harshan, J. J. Chungath and N. Ashok

ROLE OF FECUNDITY GENES IN PROLIFICACY OF SMALL RUMINANTS 34

Asha Abraham and Naicy Thomas

ECONOMIC ANALYSES OF THREE DIFFERENT METHODS OF PREGNANCY DIAGNOSIS IN DAIRY CATTLE OF KERALA 38

Arun Kurian, Josephine Francis and Jiss Job

PARTICIPATION OF SELF HELP GROUP MEMBERS AND NON- MEMBERS IN PANCHAYATI RAJ SYSTEM A COMPARATIVE STUDY 42

Anu George, P. J. Rajkamal and R.S. Jiji

10. THERMAL STRESS IN DAIRY CATTLE 45

A. Prasad, E.M. Muhammed, A. Kannan and T.V. Aravindakshan

CLINICAL REPORTS

52 - 60

11. SUCCESSFUL MANAGEMENT OF CHYLOTHORAX IN A DOG- A CASE REPORT G. Vijayakumar, S. Sivaraman, E. Venkatesakumar and M. Subramanian

Vol. 10 Issue 3 December 2012 JIVA <03 <

CONTENTS 12. SURGICAL MANAGEMENT OF TESTICULAR SEMINOMA A CASE REPORT 55

Laiju. M. Philip and M. Ranjith Mohan

13. A REPORT ON THE OCCURANCE OF Hymenolepis anatine IN DUCKS UNDER BACKYARD SYSTEM G. Jyothimol, K. Syamala, M.N. Priya, C.K. Deepa, K.G. Ajithkumar, Ajith Jacob George and Reghu Ravindran

14. MANAGEMENT OF SUBCUTANEOUS EMPHYSEMA IN A HORSE 59

Mir Aamir Ali and H.S. Mahesha

GENERAL ARTICLES

61 - 77

15. IMPACTS OF CLIMATIC CHANGE IN ANIMAL HUSBANDRY- ARE WE PREPARED? 61

S.S. Rani

16. VACCINES FROM OUR GARDEN 66

Tincy Mary John, N.M. Shah, B.S. Chandel and H.C. Chauhan

17. BIODIESEL PRODUCTION FROM ANIMAL FATSAN EVER GREEN TECHNOLOGY FOR THE FUTURE ENERGY SECURITY 72

John Abraham and Ramesh Saravana Kumar

ASSOCIATION NEWS

78 - 80

The editor/editorial board and referees are in no way responsible individually or collectively for the views, data and technical details presented in the contributed papers

J. Ind. Vet. Assoc., Kerala. 10 (3) <04 <

RESEARCH ARTICLE

BETA CASEIN A1A2 POLYMORPHISM AND MILK YIELD IN VECHUR, KASARGODE DWARF AND CROSSBRED CATTLE* E. M. Muhammed1 and M. Stephen2 College of Veterinary and Animal Sciences, Mannuthy, Thrissur *Part of MVSc thesis of the first author submitted at Kerala Agricultural University

ABSTRACT â-Casein (â-CN) is the second most abundant protein in cow's milk that contains 209 amino acids. Among the twelve â-CN variants identified A1 and A2 are the common types and others are very rare. A1 variant has histidine at position 67 of the amino acid sequence while A2 possess proline at this position- this polymorphism has attracted much public health attention, since consumption of A1 milk is reported to cause various illness. The high producing Bos taurus cattle are found to possess more A1 allele. The present study was undertaken to assess the relationship between â-CN A1/A2 polymorphism with milk yield in Vechur, Kasargode and Crossbred cattle. The average daily milk yield was 1.27±0.05 kg in Vechur cattle and the peak yield of crossbred cattle was 10.71±1.046 kg. The average daily milk yield for A1A2 genotype in Vechur cattle was 1.52±0.08 kg and 1.14±0.04 kg for A2A2 genotype and the difference in the milk yield of the two genotypes was significantly different 1* 2

PhD scholar, Email:dremmuhammed@gmail.com

Professor at Department of Animal Breeding, Genetics and Biostatistics, College of Veterinary and Animal Sciences, Mannuthy, Thrissur

(p=0.05). In crossbred cattle the peak yield (kg) of A1A1 genotype was 14.64±3.181, 8.54±0.194 for A1A2 genotype and 9.09±0.125 for A2A2 genotype. The A1A1 genotype in crossbred cattle showed significantly higher average peak yield compared to other genotypes and difference between A1A2 and A2A2 genotypes were not significantly different (p=0.05). Thus it can be concluded that selection for enhancing milk production may increase the frequency of harmful A1 allele in bovine population. Key words: Beta casein gene A1and A2, Vechur cattle, AS-PCR, Milk yield.

INTRODUCTION India has vast resources of livestock. It plays a vital role in the Indian economy and in improving the socio economic conditions of rural masses. Agriculture and livestock sector share 16.74 percent and 4.36 percent in the economy of India. As per Livestock census 2003, India has total livestock population of 485 million with 185.2 million cattle and 97.2 million buffalo which are 65 percent and 34 percent respectively of total bovine population (283.1million). In Kerala 96 percent of livestock population is cattle producing 98.50 percent of total milk (Anon, 2009). More Vol. 10 Issue 3 December 2012 JIVA <05 <

than 82 percent of the cattle in Kerala are crossbreds except indigenous breeds like Vechur. Vechur cattle once thought to be extinct due to extensive crossbreeding are the first native cattle of Kerala to be approved as a distinct breed and the smallest breed in the world. Vechur cattle are maintained by a few farmers and at Kerala Veterinary and Animal Sciences University. Milk is a common source of animal protein and associated micro elements for vegetarians. Cow's milk contains two major protein groups: caseins and whey proteins and out of which caseins account for 80 percent of milk proteins. Bovine milk contains four caseins: alpha S1 (CSN1S1 39-46 percent of total casein), alpha S2 (CSN1 S2 8-11 percent), beta (CSN2, 25-35 percent), and kappa (CSN3 8-15 percent). âCasein (â-CN) is the second most abundant protein in cow's milk that contains 209 amino acids. Bovine â-CN gene belongs to the cluster of four casein genes located on chromosome 6. There are 12 genetic variants of â-CN: A1, A2, A3, B, C, D, F, H1, H2, I and G out of which A1 and A2 are the most common. So the present study was undertaken, realising the importance of â-CN gene polymorphism in discovery of markers linked to economically important traits such as milk production.

MATERIALS AND METHODS Blood samples were collected from 72 Vechur cattle (60 from KVASU farm and 10 from Vechur Conservation Trust, Vechur), 14 Kasargode dwarf cattle (KVASU farm) and 100 crossbreds from different parts of Kerala viz, Kozhikode (22), Kannur (15), Malappuram (14), Wayanad (15), Thrissur (24) and Kottayam (10). <06 < J. Ind. Vet. Assoc., Kerala. 10 (3)

Collection of Samples About 5 ml of blood was collected from each animal in a sterile 15 ml polypropylene centrifuge tube (vacutainer) containing Ethylene Diamine Tetra Acetic acid (EDTA) as anticoagulant (1 mg/ml of blood). The samples were brought to the 0

laboratory at 4 C and stored at -20 C until DNA extraction.

AS-PCR amplification There are 12 alleles (A1, A2, A3, B, C, D, F, G, H1, H2, I and G) of â-CN in bovines. Of all the 12 â-CN variants all except A2 produce BCM-7 upon enzymatic digestion in the gut. Among BCM-7 yielding variants A1 is the predominant type and other alleles are very rare. Therefore in this study alleles are classified as A1 and A2 only. Based on this, two allele specific reverse primers each one matched to one of two alleles at 3' end (G for â-CN A2 and T for â-CN A1) along with a common forward primer were used (Keating et. al., 2008). The ASPCR products were checked electrophoretically using 2 percent agarose.

Statistical analysis The allelic and genotype frequencies at A1/A2 locus were calculated by direct counting method and the variations of the allelic frequencies among the three groups were analyzed by the Chi-square test of significance as described by Snedecor and Cochran (1994) considering the allelic frequencies in a 2×2 table.

Collection of data The average daily milk yield of Vechur cattle was collected from the records maintained

in the farm. For crossbred animals, the peak yield was obtained from the farmers. The milk yield of Kasargode dwarf cattle were not recorded as the number of observations was very less. There were only two genotypes (A1A2 and A2A2) in Vechur cattle and three genotypes- A1A1, A1A2 and A2A2 in crossbred cattle. The difference in milk yield of the genotypes was compared by t-test and ANOVA.

RESULTS AND DISCUSSION Genotype and Allele Frequencies of â-CN The allele and genotype frequencies of âCN A1A2 in Vechur, crossbred and Kasargode dwarf cattle are presented in Table 4.1. In Vechur cattle the genotype frequencies of A1A1, A1A2 and A2A2 were observed as 0, 0.34 and 0.66 whereas in crossbred cattle the corresponding frequencies were 0.32, 0.28 and 0.40 respectively. Of the 72 Vechur cattle typed, none of the animals were of A1A1 genotype. No A1A1 genotypes were found in Kasargode dwarf animals also and the corresponding frequencies in Kasargode dwarf cattle was recorded as 0.79 (A1A2) and 0.21 (A2A2). The frequencies for A1 and A2 alleles in Vechur cattle were noted as 0.2 and 0.80 respectively and for crossbreds the frequencies were recorded as 0.46 and 0.54, respectively. In the Kasargode dwarf cattle A1 and A2 allele frequencies were obtained as 0.39 and 0.61 respectively.

â- CN Polymorphism and Milk Yield The average daily milk yield of 50 Vechur cows were found to be 1.27±0.07 kg and the average peak yield of 100 crossbred cattle were

Where a, b, c are allelic frequencies in different groups

found to be 10.71±1.05 kg. â-CN A1/A2 polymorphism had significant effect on average daily milk yield in the Vechur cattle and peak yield in crossbred cattle (Table 4.2). In t test, Vechur cattle carrying A1A2 genotype showed a significantly higher average daily milk yield (1.52±0.08 kg) compared to A2A2 genotype (1.14±0.04 kg). ANOVA showed significant difference between two genotypes in crossbreds. Crossbred cattle carrying A1A1 genotype showed significantly higher average peak yield (14.64±3.181 kg) whereas A1A2 and A2A2 genotypes had lower average peak yield (8.54±0.019 kg and 9.09±0.125 kg respectively). The peak yields of A1A2 and A2A2 were not significantly different while the peak yield of A1A1 genotype was significantly higher than the other two genotypes. Vol. 10 Issue 3 December 2012 JIVA <07 <

The recorded average daily milk yield of Vechur cattle was 1.27±0.07 kg in the present study against 2.17 kg reported by Girija (1994), 2.2 kg reported by Iype and Venketachelapathy (2001). In crossbred cattle the peak yield was recorded as 10.71±1.046 kg. The 305 day milk yield of crossbred cattle of Kerala is reported as 2406 kg by Chandran and Stephen (2007) and 2106 kg by Anand (2009). Significantly increased level of milk yield obtained in A1A1 crossbred cattle is in line with findings of Ron et.al. (1994) in Israeli Holstein cows and Velmala et.al. (1995) in Finnish Ayrshire cattle. Higher milk production from A2 allele is reported by Ng-Kwai-Hang et.al.(1990) in Holstein Friesian, Ikonen et.al. (1999) in Finnish Ayrshire, Freyer et.al. (1999) in Holstein Friesian, Comin et.al. Means with different superscript in a row differ significantly (p=0.05) Figures in parenthesis denotes number of observations (2008) in Italian Holstein Friesian and Nilsen et.al. (2009) in Norwegian Red cattle.A higher milk yield was reported for heterozygous A1A2 animals by NgKwai-Hang et.al. (1986) and Ojala et.al. (1997) in Holstein Friesian cattle. In contrast, milk yield was not associated with â-CN A1/A2 polymorphism as reported by Sarbour et.al. (1996) in Canadian Holstein and Boettcher et.al. (2004) in Holstein Friesian cattle. As observed in the present study, selection for increasing milk yield may contribute for the higher proportion of undesirable A1 alleles in the population (Ron et.al. 1994 and Velmala et.al. 1995). Considering the public health implication, adequate weightage should be given to select bulls with A2A2 genotype while making selection for increasing milk yield of crossbreds. The proportion <08 < J. Ind. Vet. Assoc., Kerala. 10 (3)

of A2 allele is high in Vechur cattle. Based on the present results, careful breeding strategies can be adopted to develop a Vechur herd/breed with A2A2 genotype and its milk can be marketed at a premium price.

ACKNOWLEDGEMENTS The authors are thankful to the Dean, College of Veterinary and Animal Sciences, Mannuthy, Director, Department of Animal Breeding, Genetics and Biostatistics, College of Veterinary and Animal Sciences, Mannuthy, Thrissur, Kerala and Vechur Conservation Trust, Vechur, Kerala, for providing facilities for the study.

REFERENCES Anand, L. F. 2009. Leptin gene polymorphism in Vechur and Crossbred cattle of Kerala. MVSc. Thesis, Kerala Agricultural University, Thrissur, p. 79. Anon. 2009. Annual report, Department of Animal Husbandry and Dairying, Government of India, New Delhi. Available:http:// www.dahd.nic.in.[15 Feb 2010]. Boettcher, P.J., Caroli, A Stella, A., Chessa, S., Budelli, E., Canavesi, F., Ghiroldi, S. and Pagnacco, G. 2004. Effects of Casein Haplotypes on Milk Production Traits in Italian Holstein and Brown Swiss Cattle. J. Dairy Sci. 87:4311-4317. Chandran, R. R and Stephen, M. 2010. Evaluation of lactation milk yield in crossbred cattle. Ind. Vet. J. 87:363-364. Comin, A., Cassandro, M., Chessa, S., Ojala, M., Dal Zotto, R., De Marchi, M., Carnier, P. and

Gallo, L. 2008. Effects of composite 창-casein genotypes on milk coagulation, quality and yield traits in Italian Holstein cows. J. Dairy Sci. 91:4022-4027 Freyer, G., Konig, S., Fischer, B., Bergfeld, U. and Cassell, B.G. 2008. Crossbreeding in dairy cattle from a German perspective of the past and today. J. Dairy. Sci. 91:3725-3743. Freyer, G., Liu, Z., Erhard, G.T. and Panicke, L. 1999. Casein polymorphism and relation between milk production traits. J. Anim. Breed. Genet. 116: 87-97. Girija, C.R. 1994. Characterization and evaluation of dwarf cattle of Kerala, Ph.D. Thesis, Kerala Agricultural University, Thrissur. p. 112. Ikonen, T., Ojala, M. and Ruottinen, O. 1999. Associations between milk protein polymorphism and first lactation milk production traits in Finnish Ayrshire cows. J. Dairy Sci. 82:1026-1033. Iype, S. and Venketachelapathy, R.T. 2001. Vechur cattle of Kerala, Kerala Agricultural University, Thrissur, p 37. Keating, F., Smith, T.J, Ross, R.P. and Cairns, M.T. 2008. A note on the evaluation of a betacasein variant in bovine breeds by allelespecific PCR and relevance to 창casomorphin. Irish J. Agric. Fd Res. 47: 99104.

Ng-Kwai-Hang, K.F., Hayes, J.F., Moxley, J.E. and Monardes, H.G. 1986. Relationships between milk protein polymorphisms and major milk constituents in Holstein-Friesian cows. J. Dairy Sci. 69:22-26. Ng-kwai-Hang, K.F., Monardes, H.G. and Hayes, J.F. 1990. Association between genetic polymorphism of milk proteins and production traits during three lactations. J. Dairy Sci. 73:3414-3420. Nilsen, H., Oslen, H.G., Hayes, B., Sehesred, E., Svendsen, M., Nom, T., Meuwissen, T. and Lien, S. 2009. Casein haplotypes and their association with milk production traits in Norwegian Red cattle. Genet. Selection Evolution 41:1-12. Ojala, M., Famula, T.R. and Medrano, J.F. 1997. Effects of milk protein genotypes on the variation for milk production traits of Holstein and Jersey cows in California. J. Dairy Sci. 80:1776-1785. Sarbour, M. P., Lin, C.Y., Lee, A. J. and. Mcalllster, A. J. 1996. Association between milk protein genetic variants and genetic values of Canadian Holstein bulls for milk yield traits. J. Dairy Sci. 79:1050-1056. Velmala, R., Vikki, J., Elo, K. and Tanila, A.M. 1995. Casein haplotypes and their association with milk production traits in the Finnish Ireshire cattle. Anim. Genet. 26:419-425.

Vol. 10 Issue 3 December 2012 JIVA <09 <

RESEARCH ARTICLE

A HISTOLOGICAL STUDY OF THE STRUCTURAL CHANGES IN THE PANCREAS OF DIABETIC RATS* B. Dhanush Krishna1 and Suguna Rao2 Department of Pathology, Veterinary College, KVAFSU, Hebbal, Bangalore *Part of MVSc. thesis submitted by the first author to KVAFSU, Bidar

ABSTRACT The present study was aimed to demonstrate the histological changes in the pancreas of streptozotocin (STZ) induced diabetes in a rat model. Twenty rats were randomly separated into two groups of ten rats each. Group I served as normal control and group II served as diabetic. The diabetic condition was induced in group II by streptozotocin. Light microscopic evaluation of islets showed highly swollen 창-cells with loss of cytoplasmic granularity, vacuolations, necrosis, elongated and fusiform 창-cells and sparse cellularity in diabetic rats. The present study showed that induction of diabetes using STZ results in the alteration of the morphology of endocrine part of pancreas in rats. Keywords: 창-cells, Diabetes Mellitus, Necrosis, Streptozotocin

INTRODUCTION Diabetes mellitus (DM) is a major health problem worldwide. The economic impacts of diabetes with its complications and associated diseases are large (Sarah et al., 2004). According to recent estimates, the prevalence of diabetes mellitus is 4 percent worldwide and that indicates 143 million persons are affected which will increase to 300 million by the year 2025 (Analava et al., 2007). Animal models in diabetes research are very common where rats are the first choice of use, comprising over 85 percent of these models (US Department of Agriculture, 1989). It may have been because of the pathogenesis of diabetes in animal models is most likely similar to the pathogenesis in humans. Streptozotocin (STZ) has been extensively used to induce diabetes for various diabetes studies in laboratory animals. STZ has been reported to be capable of generating reactive oxygen species resulting in oxidative stress and cell death. Also, STZ has been found to be a better chemical inducer for diabetes than alloxan (Szkudelski, 2001).

PhD Scholar, Department of Pathology, College of Veterinary and Animal Sciences, KVASU, Mannuthy, Email ID: dhanushkb@gmail.com 2

Professor, Department of Pathology, Veterinary College, Bangalore <10 < J. Ind. Vet. Assoc., Kerala. 10 (3)

The present study was designed to demonstrate the histological changes on the endocrine component of pancreas in STZ induced diabetes in a rat model.

MATERIALS AND METHODS Care and management of experimental animals The present study was carried on approval from Institutional Animal Ethics Committee of Veterinary College, Bangalore. Adult male Albino Wistar rats weighing 220±40g were maintained under standard laboratory conditions. The animals were randomly separated into two groups, group I as normal control, while group II as diabetic.

Induction of experimental diabetes After a two-week acclimatization period, DM was induced experimentally in group II by a single intraperitoneal injection of a freshly prepared STZ solution (Sigma Chemicals, USA) dissolved in 0.1 M ice-cold citrate buffer (pH 4.5) at a dose of 45 mg/kg to overnight-fasted rats (Babu and Prince, 2004). Control rats received an intraperitoneal injection of citrate buffer alone. After three days of STZ administration, blood glucose levels of each rat were determined. Rats with a blood glucose level above 300mg/dL were considered diabetic and included in this study. The blood glucose level was measured using commercially available biochemical kits (Span Diagnostics, Bangalore) (Tietz, 1976).

Histological Evaluation Two animals from each group were euthanized on 15th, 30th and 45th day of the experiment using an overdose of ether for studying the progressive effects of STZ. Animals were immediately dissected and pancreas was observed for evidence of gross pathology. For the light microscopic examinations, samples from tail portions of the pancreas were fixed in 10 percent

neutral buffered formalin and embedded in paraffin. The paraffin blocks were cut into pieces with a thickness of 4 µm. The sections were stained with Hematoxylin and Eosin (H&E) (Bancroft and Gamble, 2008) for the evaluation of islets injury.

Statistical Analysis Data were analyzed using a commercially available statistics software package (Graph Pad Prism for Windows Version 5.0). All values are presented as Mean ± Standard Error (SE). The data were analyzed using paired sample student t test (p < 0.001; two tailed).

RESULTS AND DISCUSSION A significant (p < 0.001) decrease in the body weight was observed in the diabetic group when compared to the control. The blood glucose level was significantly increased (p < 0.001) in the diabetic group when compared to the control. The diabetic control rats appeared grossly emaciated. The pancreas showed slight congestion and progressive decrease in the size which became th

appreciable from 15 day of the present study. On th

45 day, the pancreas was atrophied and appeared as a thin gelatinous strip. Histopathology of islets of Langerhan's of pancreas of control animals revealed normal architecture with compact arrangement of â- cells and non â- cells throughout the study. In the present study, post STZ injection, the pancreas showed numerous lobules with normal appearing exocrine component in diabetic control rats on th

15 day. However, pancreatic islets were reduced in number per lobule and showed loss of normal architecture. The normal distribution of non Vol. 10 Issue 3 December 2012 JIVA <11 <

â-cells and â-cells was altered with much reduction in the number of â-cells and disorderly arranged non â- cells. The â-cells were swollen, vacuolated, necrotic or elongated and fusiform with condensed nucleus and showed loss of cytoplasmic granularity.

to locate. There was severe reduction in the number of â-cells. Occasional persisting â-cells were highly swollen, vacuolated and showed loss of cytoplasmic granularity. This was in agreement with earlier reports. (Papaccio et al., 2000 and Mir et al., 2008). They appeared irregular in shape, reduced in the size, with some assuming 'star fish' appearance. Besides, there was an increase in the number of non â- cells and mild degrees of fibrosis with infiltration of a few inflammatory cells were also observed. The histopathology of islets in diabetic rats indicated progressive destruction of th

â-cells from 15 to 45 day of the investigation.

Fig 1A. Islet of control animal showing normal architecture with compact arrangement of th â- cells and non â- cells on 15 day (H&E X 200)

Fig 1C. Islet showing extensive vacuolation and th necrosis of â- cells on 30 day (H&E X 200)

Fig 1B. Islet of diabetic control showing atrophied th and vacuolated â- cells on 15 day (H&E X 200) th

On 30 day post STZ injection, there was further decrease in the number of islets which appeared shrunken and showed increased severity in those lesions that appeared on 15th day in â-cells. On 45th day post STZ injection, occasional small sized islets were appreciated which were difficult <12 < J. Ind. Vet. Assoc., Kerala. 10 (3)

Fig 1D. Islet showing atrophied and “starfish” appearance on 45th day (H&E X 200)

According to the American Diabetes Association (ADA), most common symptoms of diabetes include polydipsia, weight loss and polyphagia those were evidently present in the diabetic groups in the present experiment. The elevation of blood sugar level on 3rd day confirmed the establishment of diabetes mellitus in rats which is attributed to the selective cytotoxicity of STZ on â- cells (Bedoya et.al., 1996). The decrease in cellularity within islets observed in the study reflects the cytotoxity of STZ. The reduction in the number of â- cells was also noticed. STZ possesses diabetogenic effect mediated through pancreatic â-cell destruction. STZ appears to cause cytotoxicity by a number of mechanisms. STZ on entry into the â- cells via a glucose transporter (GLUT2) gets spontaneously decomposed to form an isocyanate compound and a methyldiazohydroxide which cause intra molecular carboxylation and alkylation of cellular components respectively especially that of DNA of â-cells (Varva, 1960). However, the DNA damage of â-cells of pancreas is mainly by alkylation with carbonium ion produced by methyldiazohydroxide (Elsner et.al., 2000). The DNA damage induces activation of poly ADP-ribosylation, a process that is more important for the diabetogenicity of STZ than just DNA damage itself. Poly ADP-ribosylation leads to +

depletion of cellular NAD and ATP. Enhanced ATP dephosphorylation after STZ treatment supplies a substrate for xanthine oxidase resulting in the formation of super oxide radicals. Consequently, hydrogen peroxide and hydroxyl radicals are generated. Further, STZ liberates toxic amounts of nitric oxide that inhibits aconitase activity and participates in DNA damage. As a result of the STZ action, â-cells undergo destruction by necrosis.

STZ selectively destroys the pancreatic insulin secreting â- cells, leaving less active cells and result in a diabetic state (Szkudelski, 2001). In the present study, â- cells in some islets were found to be fusiform. The change in the shape of cells can be attributed to the partial damage of STZ to some â- cells. Aybar et.al. (2001) have reported that use of lower dose of STZ produced an incomplete destruction of pancreatic â-cells even though rats became permanently diabetic. Brownlee (2001) postulated a concept that link hyperglycaemia-induced damage by different mechanisms that finally leads to cellular stress. Firstly, hyperglycemia increase movement of glucose through polyol pathway and sorbitols are produced which in turn causes osmotic stress to cells and dihydronicotine amide adenine dinuleotide phosphate (NADPH) is consumed, depleting intracellular glutathione. Secondly, hyperglycemia increases concentrations of advanced glycation end products. These glycosylated proteins are formed by nonenzymatic reactions and changes in protein structure may alter their cellular functions. Thirdly, glucose activates various isomers of protein kinase C which in turn affects the expression of nitric oxide, endothelin, nuclear factor kappa B (NF-kB) and plasminogen activator inhibitor. Finally, hyperglycemia increases the flux of glucose through the hexasomine pathway effecting inflammatory mediators and insulin resistance. The combined effect of these mechanisms results in over-production of superoxides by the mitochondrial electron-transport chain, causing cellular stress and damage that was clearly appreciated in the form of islets injury in the present study. STZ induced hyperglycaemia has Vol. 10 Issue 3 December 2012 JIVA <13 <

been described as a useful experimental model to study the activity of hypoglycaemic agents. So the result of the present study is useful to evaluate the hypoglycaemic agents in pre-clinical trials.

REFERENCES American Diabetes Association: Report of the Expert Committee on the diagnosis and classification of diabetes mellitus. 2007, Diabetes Care, 30, S42-47. Analava, M., Bhattacharya, D and Roy,S. 2007. Dietary influence on type 2 Diabetes (NIDDM). J. Hum. Ecol., 1:139-147. Aybar, M. J., Sanchez Riera, A. N., Grau, A and Sanchez, S. S. 2001. Hypoglycaemic effect of the water extract of Smallanthus soncifolius (yacon) leaves in normal and diabetic rats. J. Ethnopharmacol. 74: 125132. Babu, P. S and Prince, P. S. M. 2004. Antihyperglycaemic and antioxidant effect of hyponidd, an ayurvedic herbomineral formulation in streptozotocin-induced diabetic rats. J. Pharm. and Pharmacol., 56: 1435-1442. Bancroft, J. D and Gamble, M. 2008. Theory and th

practice of histological techniques. 6 Ed. Churchill Livingstone, United States of America, 121 p. Bedoya, F. J., F. Solano and Lucas, M.1996. Nmonomethyl-arginine and nicotinamide prevent streptozotocin induced double strand DNA break formation in pancreatic rat islets. Experientia, 52:344-347.

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Brownlee, M. 2001. Biochemistry and molecular cell biology of diabetic complications. Nature, 414: 813-820. Elsner, M., Guldbakke, B., Tiedge, M., Munday, R and Lenzen, S., 2000. Relative importance of transport and alkylation for pancreatic cell toxicity of streptozotocin. Diabetologia. 43: 1528-1533. Mir, S. H., Baqui, A., Bhagat, R.C., Darzi, M.M and Shah, A.W. 2008. Biochemical and histomorphological study of streptozotocin - induced diabetes mellitus in rabbits. Pak. J. Nutr., 7: 359- 364 Papaccio, G., Pisanti, F.A., Latronico, M.V., Ammendola, E and Galdieri, M.2000. Multiple low dose and single high dose treatments with streptozotocin do not generate nitric oxide. J. Cell Biochem., 77: 82-91. Sarah, W.,Gojka, R.,Anders, G.,Richard, S and Hilary, K., 2004. Global prevalence of diabetes. Diabetes Care.2004; 27: 10471053. Szkudelski, T. 2001. The mechanism of alloxan and streptozotocin action in cells of the rat pancreas. Physiol. Res., 50: 536-546. Tietz. 1976. Fundamentals of clinical chemistry. W.B.Saunders Co., Philadephia. US Department of Agriculture: Animal welfare enforcement report fiscal year 1988. Washington, DC, 1989. Varva, J.J., Deboer, C. and Dietz, A. 1960. Streptozotocin, a new antibacterial antibiotic. Angtibiot. Ann. 230-235.

RESEARCH ARTICLE

THERAPEUTIC POTENTIAL OF AFANIL® AGAINST BLOAT AND FOR EARLY RESTORATION OF RUMEN FUNCTION IN BOVINES S. U. Digraskar, V. D. Muley, K. Ravikanth, M. Dandale and S. Maini College of Veterinary and Animal Science, MAFSU, Parbhani, Maharastra and Ayurvet Limited, Baddi, Himachal Pradesh.

ABSTRACT Efficacy of herbal oral formulation ®

Afanil (Supplied by M/s Ayurvet Ltd, Baddi, HP; India) was evaluated against bloat condition in bovines on the basis of improvement in clinical, hematological and rumen fluid analysis. For this a clinical study was undertaken on 20 cases of bovines suffering from bloat presented to Department of Veterinary Medicine, College of Veterinary and Animal Sciences, Parbhani, Maharashtra. Total cases were divided in two treatment groups T1 and T2 of 10 animals each. T1 was administered conventional antizymotic mixture of vegetable oil 750 ml-1.5 liter mixed with 20-30 ml of turpentine oil orally. T2 was treated with herbal anti bloat oral formulation (Afanil®) @ 100 ml twice daily till recovery. Supportive and fluid therapy was also given to the animals from both the groups. Time period required in hours for resumption of distended rumen and rumination process was significantly ®

lesser (P < 0.01) in animals treated with Afanil (6.08±0.37 and 6.95±0.39) as compared to conventional anti bloat therapy in T1 (9.65±0.71 and 10.75±0.71). 2.50± 0.17 and 1.70± 0.15 days was the mean time period required for complete clinical recovery after conventional

and Afanil® anti-bloat therapy respectively.

Afanil® efficiently normalized the altered hematological as well as rumen pH conditions as compared to that of T1. In present clinicotherapeutic trial, Afanil® herbal anti bloat preparation was found to be effective to treat bloat cases of bovines in comparison to conventional therapy. It contains herbal extracts and carminative volatile oils which ensure quick and prompt relief in bloat cases hence may be successfully employed in the treatment. ®

Keywords: Afanil , bloat, antizymotic, tympany, herbal

INTRODUCTION Bloat is an over distention of the rumen and reticulum with the gases of fermentation either in the form of a persistent foam mixed with the ruminal contents called primary or frothy bloat, or in the form of free gas separated from the ingesta called secondary or free-gas bloat (Merck Veterinary Manual, 2011). It occurs due to feed rations containing a high proportion of grains or due to feeding of fresh green legumes. In acute tympany or bloat, due to progressive distension of rumen there is respiratory and cardiac distress which if not relieved, may rapidly lead to asphyxia, cardiac failure and death (Radostits et. al, 2003). Vol. 10 Issue 3 December 2012 JIVA <15 <

Comparative efficacy of the Afanil® herbal anti bloat therapy with that of conventional therapy was evaluated in the present clinical study. MATERIALS AND METHODS The present investigation was undertaken at Department of Veterinary Medicine, College of Veterinary and Animal Sciences, Parbhani, MAFSU, Maharashtra. Twenty bovine cases presented with clinical features of frothy as well as free gas bloat formed the basis of the experiment. The clinical cases were divided in two treatment groups T1 and T2 of 10 animals each. Feeding and clinical history was recorded in every case which followed blood and ruminal fluid sample collection for further analysis. Animals in group T1 (n=10) suffering from bloat of dietary origin were administered conventional antizymotic mixture of vegetable oil 750 ml-1.5 liter mixed with 20 -30 ml of turpentine oil orally. Group T2 (n=10) was treated with herbal ®

anti bloat oral formulation Afanil 100 ml twice orally daily till recovery (not more than 2 days in this experiment). Supportive therapy with antihistaminic, IV fluids (RL/DNS) along with sodium bicarbonate or ascorbic acid was also instituted depending upon the cause of bloat in affected bovine. Clinical signs were recorded after treatment. The efficacy of the treatment in both groups was assessed on the basis of improvement in, a) Clinical parameters e.g. time period required for resumption of distended rumen, rumination and other physiological processes. b) Hematological- TLC, DLC and c) Rumen fluid analysis- pH and consistency.

<16 < J. Ind. Vet. Assoc., Kerala. 10 (3)

RESULTS AND DISCUSSION Diagnosis was made on basis of the correlating the feeding history with that of clinical signs exhibited by the animals. Most of the animals had clinical features comprised of over distended abdomen with bulging of upper left flank, anorexia, absence of rumination and retarded ruminal motility, dehydration, dyspnoea, dullness and depression. Tympanic or drum-like sound was heard on percussion of left paralumbar fossa. Milk yield was greatly reduced or absent in lactating animals. All of the animals were having the feeding history of highly succulent leguminous fodder, flour or grains. A prompt response to treatment with ®

Afanil was noticed. Over distended rumen started getting rid of gases and froth within minutes and minimum time period required for rumen to completely assume its normal shape was only 4.50 hrs after the initiation of therapy (Table 1). While that for conventional treatment required 6.5 hrs. Mean time period required for resumption of distended rumen after treatment in Afanil® treated group was 6.08±0.37 hrs against 9.65±0.71 hrs required for conventional therapy. Similar findings were also noted by Gahlaut (1998). Mean time period required for resumption of rumination process (Table 1) after Afanil® therapy was 6.95±0.39 hrs therapy. In comparison to this 10.75±0.71 hrs were required with conventional therapy. A complete clinical recovery occured in mean time period of ®

1.70±0.15 days after Afanil therapy while conventional therapy required more time for complete recovery i.e. 2.50±0.17 days. Blood parameters with reference to TLC and DLC of affected bovines before and after

Table 1. Mean values of clinical parameters in different treatment regimen against bloat in bovines.

Group T1 Conventional therapy

Clinical parameters

Sr.No

Period required for resumption of distention of rumen in hours after treatment (in hours)

9.65± 0.71a

6.08±0.37b

10.75± 0.71a

6.95± 0.39b

2.50± 0.17a

1.70± 0.15b

Period required for resumption of rumination process in hours after therapy (in hours)

Group T2 ® Afanil therapy

Period required for overall clinical recovery e.g. normal appetite, physiological parameters and milk (in days)

Means bearing different superscript in a row differ significantly at P=0.05 leve

Table 2. Evaluation of pH of ruminal fluid in clinical cases of bloat before and after treatment. ®

Group T1 (Conventional therapy)

Group T2 (Afanil therapy)

Case no.

Mean ± SE

7.87± 0.43

7.15± 0.23

7.47± 0.58

6.77± 0.15

BT: Before treatment AT: After treatment Afanil ® therapy were studied. Differential Leukocyte Count (DLC) showed eosinophilia to the extent of 5.70±0.37 in affected cases which were improved to 3.0±0.26 and 2.70±0.21 in group T1 and T2 animals respectively. As ruminal and blood plasma histamine concentrations increase during ruminal lactic acidosis (Asenbach and Gabel, 2000) eosinophilia may be because of histamine upsurge in bloat cases which was significantly brought down in Afanil® treated group.

Analysis of ruminal fluid before and after treatment in both the groups was done (Table 2). ®

Results obtained after analysis showed that Afanil efficiently brought the altered levels of the ruminal pH to normal level required for optimum ruminal fermentation and fiber digestion.

The carminative, antispasmodic, antiflatulent, stomachic, alterative and laxative properties of Afanil® are contributed by its constituent herbs for which these activities have been well documented (Nadkarni, 1954; Chopra et. al, 1982; Kapoor, 1990; Hussain et. al 1992). Vol. 10 Issue 3 December 2012 JIVA <17 <

In the present clinico-therapeutic trial, the 速

product Afanil , herbal anti bloat preparation was found to be safe, effective, non-toxic and promising therapy to treat frothy as well as free gas bloat cases of bovines in comparison with conventional therapy. Results in the present experiment corroborates well with earlier findings of Sharma (1996), Gahlaut (1998) and Bhardwaj (1998). Thus, it was concluded that Afanil速, a liquid emulsion containing herbal extracts and carminative volatile oils ensures quick and prompt relief in bloat cases and may be successively used in field conditions to prevent the losses from bloat origin. ACKNOWLEDGEMENT Authors are thankful to Associate Dean, COVAS, Parbhani for providing necessary facilities required for the present Clinico-therapeutic trial and Ayurvet Limited, Baddi for providing necessary samples for carrying out present research experiment. REFERENCES Aschenbach, J.R and Gabel, G.J. 2000. Effect and absorption of histamine in sheep rumen: Significance of acidotic epithelial damage J. Anim. Sci., 78:464-470

<18 < J. Ind. Vet. Assoc., Kerala. 10 (3)

Bhardwaj, U. 1998. Therapeutic efficacy of herbal anti-tympanitic agents for recurrent tympany in buffaloes. Ind. Vet. Med. J. 22: 327-328 Chopra, R.N., Chopra, I.C., Handa, K.L and Kapoor, L.D. 1982. Indigenous drugs of India. 2 n d edn. Academic Publishers, Calcutta. 速

Gahlaut, K.S.1998. Therapeutic efficacy of Afanil for ruminal tympany. Dairy Guide, AprilJune, 57-59.

Hussain, A. 1992. Dictionary of Indian Medicinal Plants. Central Institute of Medicinal and Aromatic Plants, Lucknow. Kapoor, L.D. 1990. CRC Handbook of Ayurvedic medicinal plants. CRC Press, Boca Raton, Florida. Merck Veterinary Manual. 2011. http://www. merckvetmanual.com/mvm/index.js? cfile=htm/bc/ Accessed 22/7/2012 Nadkarni, A.D. 1954. Dr. K.M. Nadkarni's Indian Materia Medica. Popular Book Depot, Bombay. Radostits,M., Gay,C.C., Blood,D.C and Hunchcliff, K.W. 2003. Veterinary Medicine. 10th Ed. W.B. Saunders Company Ltd., 493-500. Sharma, V and Sharma, S.D. 1996. Clinical efficacy 速

of Afanil in treatment of acute tympany and bloat. The Veterinarian, 20:18-19.

RESEARCH ARTICLE

OPTIMIZATION OF PCR THROUGH MANIPULATION OF CYCLE TIMES AND INCLUSION OF FORMAMIDE S. Pramod1, A. P. Usha2, T. Venkatachalapathy3 and K.C. Raghavan4 Kerala Veterinary and Animal Sciences University

ABSTRACT Polymerase chain reaction (PCR) is an in vitro technique to produce million fold copies of a particular segment of DNA. PCR should optimally yield a unique product unless designed otherwise. Artefacts appear many times, which affect the success of downstream applications. Prevention of non specific amplification using formamide and manipulation of cycling time is discussed in this article.

INTRODUCTION Polymerase chain reaction (PCR) is an in vitro cell free technique to produce a large number of copies of a specific segment of DNA. The technique of PCR (Mullis, 1983) evolved during four decades after the establishment of double stranded helix structure of DNA by Watson and Crick (1953). Kleppe (1971), an associate of Hargobind Khorana probably introduced the concept of PCR; with two primers, and cycles of repair synthesis after addition of DNA polymerase in each round. Earlier PCR relied upon DNA polymerase I enzyme of Escherichia coli or rather 1

PhD Scholar, CASAGB, Mannuthy, 2Professor & Head, CPPR, Mannuthy, 3Associate Professor & Head, University Goat Farm, Mannuthy, 4Director, CASAGB, Mannuthy

its 'Klenow fragment', which was thermolabile and needs to be added afresh before each cycle. Chien et.al. (1976) purified DNA polymerase from Thermus aquaticus (EC 2.7.7.7), which later became famous as 'Taq' polymerase. The molecule had a weight in the range of 63-68 KDa with an optimum temperature of activity at 80째C in a buffer of pH of 8.0. Ever since Taq polymerase was included in PCR eliminating the need to add DNA polymerase before initiation of each cycle, it gained acceptance and was used for a myriad of downstream applications. Even though the PCR optimally should yield a unique product (unless designed otherwise) many times unintended artefacts also appear. It becomes important to avoid such spurious amplifications so as to improve the success rate of downstream applications like molecular cloning, PCR-RFLP etc. This article attempts to throw light into the optimisation of PCR, when multiple spurious amplicons appear along with the expected product.

MATERIALS AND METHODS PCR reactions were set up so as to contain 20-100 ng of template DNA, 5 nM each of forward and reverse primers (Sigma), 200 mM of each dNTP's (Fermentas), 2.5 mM of MgCl2 (Sigma), Vol. 10 Issue 3 December 2012 JIVA <19 <

0.75 U Taq polymerase (Sigma), polymerase buffer (Sigma) to a final concentration of 1X, diluted with autoclaved Millipore water in a volume of 10 µL which was amplified in a Bio-Rad thermal cycler. 'Gradient' is an option in most thermal cyclers for optimization of cycling conditions. The positions for reaction tubes are arranged rows and columns. The positions in a row can be programmed to maintain one particular temperature and is called a block. When gradient option is set, the temperature remain constant in a row and varies along a column of tube positions. The programme used for gradient PCR was as follows; initial denaturation at 94°C for three minutes, followed by 35 cycles of denaturation at 94°C for one minute, annealing temperature over the gradient for 30 seconds and extension at 72°C for 1 minute with a final extension time of 2 minutes at 72°C. The gradient used in this experiment is presented in table (1). Sl No

Block

Temperature (°C) FUT1

HSD 17â

65.0

59.0

64.3

58.4

63.2

57.4

61.4

55.8

59.3

53.9

57.7

52.5

56.6

51.5

56.0

Table 1: Block temperatures in gradient PCR The loci under study were segments from Fucosyl transferase 1 (FUT1) and 17â Hydroxy steroid dehydrogenase (HSD17â) genes, which are candidate genes (Spotter and Distl., 2006) for litter <20 < J. Ind. Vet. Assoc., Kerala. 10 (3)

traits in pigs. The primer pair FUT1 F: 5'GCCGCCACCTCTGTCTGACC-3' and FUT1 R: 5'TACCCCCTGGGCCTCTTGCC-3' were designed from Genbank accession no. L50534.1 using Primer-Blast tool (available at http://www.ncbi. nlm.nih.gov/tools/primer-blast/index.cgi? LINK_LOC=BlastHome). The forward and reverse primers had a length of 20 bases each with a GC content of 70 percent. The melting temperature (Tm) of forward primer was 65.23°C and that of reverse primer was 66.0°C. The Tm of an oligonucleotide is the temperature at which half of the molecules remain free of secondary structures. The expected product size for this amplicon was 578 bp. A segment of HSD 17â gene was amplified using the primers described by Jacobs et.al. (2002). The nucleotide sequence of forward primer was 5'- CTCCCACCCCACCTGTTC-3' and that of the reverse primer was 5' CCGTTCACCACCCCTCCTC3', with an expected product size of 273 bp. The forward primer was 18 nucleotides long while the reverse primer had a length of 19 nucleotides. The GC content was 66.67percent and 68.42 percent respectively for the forward and reverse primers. The melting temperature (Tm) of forward primer was 59.24°C while that of the reverse primer was 61.96°C. Gradient PCR was done across a block temperature of 56°C and 65°C for FUT1 locus and 50 - 59°C in the case of HSD17â locus. The amplified products were loaded into 2 percent agarose (SRL labs) gel with pre incorporated ethidium bromide (SRL labs) and electrophoresed in Tris acetate EDTA buffer at 85 volt for 45 minutes before being photographed in Geldoc (Pharmacia). The output of gradient PCR for FUT1

is presented in figure (1) and that of HSD 17â in figure (2). Several non specific products could be observed in both the loci. In order to suppress spurious amplifications, formamide was introduced into the reaction mix, along with the reduction of extension time. The composition of reaction mixture was modified by including 0. 75 percent V/V of formamide (USB) and was used to amplify FUT1 (Fig: 3) and HSD 17 â loci (Fig: 4). However mere addition of formamide did not remove all non specific amplicons and a new PCR programme was used in case of HSD 17 â gene (Table 2).

V/V) incorporated in the reaction mixture using the modified programme. Problem of spurious amplifications could be successfully controlled in FUT1 locus (Fig: 3) and an annealing temperature was selected for subsequent amplification of this locus. In case of HSD 17 â locus, four non specific products, all above the intended product of 273 bp was observed. Addition of formamide and reduction of extension and annealing times did not quite well remove non specific amplifications (Fig: 5, lanes 5 to 7). Annealing time was reduced to 15 seconds and the extension time was reduced to 30 seconds. The PCR program was further modified by introducing an additional step with three cycles, at a higher annealing temperature and the spurious amplification could be suppressed (Fig.4).

DISCUSSION Table 2: Modified PCR programme used for HSD 17â locus

RESULTS The result from gradient PCR of FUT1 locus is given in Fig (1) and that of HSD 17 â is provided in Fig (2). Four non specific products lower in molecular mass than the expected amplicon (578 bp) were observed in case of FUT1 locus. A product in excess of 1 Kb also appeared when the annealing temperature was below 59.3°C. The larger product failed to amplify when the annealing time was reduced from 30 seconds to 15 seconds, with the simultaneous reduction of extension time from one minute to 45 seconds. A second gradient PCR was performed with formamide (0.75 percent

Efficiency of PCR reflects the capability of a primer pair to produce faithful amplification, which is close to the theoretical doubling of target sequence per cycle. Ill designed primers allow coamplification of unintended regions compromising both specificity and efficiency. The principles of PCR primer design were reviewed by Singh and Kumar (2001). However in many cases, whole genome sequence is not available in public domain which makes the researcher to choose primers without the knowledge of other probable targets. The annealing temperature (Ta) used for primer- template interaction is a function of Tm. The annealing temperature occurs within a range of 4-10°C lower than Tm and increased Ta in PCR results in poor or no amplification (Wu et.al., 1991). In case of both FUT1 and HSD 17â, the Vol. 10 Issue 3 December 2012 JIVA <21 <

annealing temperature was observed to be within this range. It implies that the nearest neighbour thermodynamics (Santa Lucia.,1998) can efficiently predict the Tm. Lorenz (2012) suggested that the melting temperature of the primers should range between 45-65°C with the window between 5258°C being the optimum. However Tm for the primer pair should not differ by more than 5°C. Dieffenbach et.al., (1993) were of the opinion that PCR primers should maintain a reasonable GC content. Oligonucleotides which were 20 bases long with a 50percent G + C content generally had Tm values in the range of 56-62°C. Primers for both the loci were within the length limit and had acceptable Tm. Even with GC content in excess of 50 percent, spurious amplicons had appeared in the PCR for FUT1 and HSD 17â loci. The high GC content is reflected in the elevated Tm of the primers. The factor that the length of primers for HSD 17â being slightly less (18 and 19 bases) than the optimum, could also have favoured mispriming. The concentration of MgCl2, dNTP's and Taq polymerase also influence the occurrence of spurious amplicons. Higher concentration of MgCl2, dNTP and Taq polymerase is known to favour mispriming and generation of unintended products. The recommended concentration of MgCl2 varies between 0.5 to 5.0 mM. In this experiment, a concentration of 2.5mM MgCl2 was used which is well within this range. Sarkar et.al. (1990) used formamide to prevent spurious amplification in PCR, which employed primers with GC content in excess of 55 percent. They had further observed that in case of primers with high GC content, formamide added to the reaction mixture at 1.5 to 5 percent levels <22 < J. Ind. Vet. Assoc., Kerala. 10 (3)

prevented spurious amplification. In the present study, formamide included at 0.75 percent V/V, was sufficient to banish non specificity. Inclusion of a separate cycle with three steps (steps 2 to 4, Table: 2) to amplify HSD 17â comes from 'Touchdown' programme (Don et.al., 1991). The touchdown programme is widely used to amplify repetitive sequences, like microsatellite markers which generate lot of noise. In order to overcome spurious priming, few initial cycles are run at higher annealing temperature thereby enhancing the specificity of primer binding. Addition of a high temperature step in this experiment possibly increased the specificity of PCR. Innis et.al. (1988) observed that Taq polymerase added nucleotides at a rate of about 60 nucleotides per second. Some other reports suggest the speed to be between 35 and 100 nucleotides per second. The rule of the thumb is to provide extension time of one minute per Kb size of the product. In current study, reduction of extension time also played a part in blocking spurious amplifications in both FUT1 and HSD 17â loci.

CONCLUSION Non specific amplification in the case of primers with high GC content could be averted by using formamide and adjustment of annealing and extension time and introduction of a high temperature step so as to favour the formation of the product of interest.

ACKNOWLEDGEMENTS INSPIRE fellowship provided by Department of Science & Technology,

Government of India, funds and facilities provided by KVASU and ICAR is thankfully acknowledged.

REFERENCES: Chien, A., D.B. Edgar and J.M. Trela. 1976. DNA polymerase from the extreme thermophile Thermus aquaticus. J. Bact., 127 (3): 15501557. Dieffenbach, C. W., Lowe, T.M and Dveksler, G.S. 1993. General concepts for PCR primer design. Genome Res.,3: S30-S37. Don, R.H., P.T. Cox, B.J. Wainwright, K. Baker and Mattick, J.S. 1991. 'Touchdown' PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res. 19(14): 4008. Innis, M.A., B. K.B. Myambo, D.H. Gelfand, and Brow, M.A.D. 1998. DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA. Proc. Natl. Acad. Sci. 85: 9436-9440. Jacobs, K.., M. Mattheeuws, , M. Van Poucke, A. Van zeveren and Peelman, l.J. 2002. Characterization of the porcine FABGL gene. Anim. Genet. 33: 220-223. Kleppe, K., E. Ohtsuka, R. Kleppe, I. Molineux and H.G. Khorana. 1971. Studies on polynucleotides. XCVI. Repair replications

of short synthetic DNA's as catalyzed by DNA polymerases. J. Molec. Biol. 56: 341-61. Lorenz, T. C. 2012. Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies. J. Vis. Exp. (63), e3998, DOI: 10.3791,3998. SantaLucia, J. Jr. 1998. A unified view of polymer, dumbbell, and oligonucleotide DNA nearest-neighbor thermodynamics. Proc. Natl. Acad. Sci. U. S. A. 95: 1460-1465. Sarkar, G., S. Kapelner and Sommer, S.S.1990. Formamide can dramatically improve the specificity of PCR. Nucleic Acids Research. 18(24): 7465. Singh, V.K. and Kumar. A. 2001. PCR primer design. Molecular Biology Today, 2 (2): 27-32 Spotter, A. and Distl, O. 2006. Genetic approaches to the improvement of fertility traits in the pig. Vet. J., 172: 234-247 Watson, J.D and Crick, F.H.C. 1953. A structure for deoxyribose nucleic acid. Nature. 171: 737738 Wu, D., L. Ugozzoli., B.K. Pal., J. Qian and Wallace, R.B. 1991. The effect of temperature and oligonucleotide primer length on the specificity and efficiency of amplification by the polymerase. DNA and cell biology, 10 (3): 233-238

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RESEARCH ARTICLE

MILK FEEDING STRATEGIES FOR IMPROVING GROWTH PERFORMANCE IN CROSSBRED DAIRY CALVES 1

A.J. Flamy and Joseph Mathew

College Of Veterinary and Animal Sciences, Mannuthy, Thrissur

ABSTRACT The success of dairy industry depends to a large extent on rearing of calves to a breedable age at a faster rate and with minimum mortality. The higher the plane of nutrition the earlier the onset of puberty and thus quicker the returns. In this context, a study was conducted to assess the effect of challenged milk feeding on growth performance in dairy calves. Eighteen calves born in Kerala Agricultural University Livestock Farm, Mannuthy were selected at random and were divided into three groups of six animals each. The calves in the first group were fed with milk in normal regime as per package of practices recommendations (Control group). Animals in the second and third group were fed 25 percent and 50 percent extra milk as that of normal regime. The body weight and average daily th

weight gain of all animals were recorded till 4 month of age and analyzed. The overall result indicated that there was a significant increase (P<0.05) in daily average weight gain in animals fed with 25 percent and 50 percent extra milk when compared to animals in the control group. It was also observed that the daily 1

Research Assistant (dr.flamy.jacob@gmail.com), 2Prof. and head, University Livestock Farm, Mannuthy, College Of Veterinary and Animal Sciences, Mannuthy, Thrissur <24 < J. Ind. Vet. Assoc., Kerala. 10 (3)

average weight gains recorded in animals in the second and third group were significantly higher (P<0.05) than that of the control during the fourth month of age indicative of the carry over effect of the feeding system. Based on the result it is recommended that a higher level of milk feeding regime during pre-weaning state will not only influence the growth rate during the period but also beneficially contribute to the growth performance in later stages. Key words: Challenged milk feeding, extra milk allowance, milk feeding strategy. INTRODUCTION An efficient calf feeding system is critical because it determines the future income and sustainability of dairy farms (Tozer et.al., 2001). The primary goal of most liquid feeding programs for dairy calves is to double the birth body weight within 8 weeks of age and minimize the morbidity primarily diarrhea and respiratory diseases and mortality ( Jasper et.al., 2000). The higher the plane of nutrition the earlier the onset of puberty and thus quicker the returns ( Von et.al., 2006). Nature's way of feeding calves includes free access, nursing until satiated, frequent meals per day and suckling. Conventional rearing systems usually limit access, restrict milk intake per meal,

encourage rapid feeding or gorging, restrict meals per day or provide milk in pails( Davis et.al.,2011 ). Feeding less milk results in poor growth due to lack of needed nutrients( Huber et.al., 1984). In many animal models, it is well documented that the amount of nutrients consumed early in life has long-term effects on future performance (Pettersson. 2000 ). In this context, a study was conducted to assess the effect of challenged milk feeding on growth performance in dairy calves reared in Kerala Agricultural University Cattle Farm.

Development Scheme, Mannuthy were selected at random and were divided into three groups of six animals each. The calves in the first group were fed with milk in normal regime as per package of practices recommendations (Control group). Animals in the second and third group were fed 25 percent and 50 percent extra milk as that of normal regime. This schedule in all groups was continued until the calves attained 3 months of age at weaning. The body weight and average daily weight gain of all animals were recorded till 4 month of age and analyzed.

MATERIALS AND METHODS

RESULT

Eighteen calves born in University Livestock Farm and Fodder Research and

The observations are summarized in the following tables

Table 1 : Details of Weight recorded (Grams) during the first, second, third and fourth month Wt. in Wt. in Wt. in Wt. in Experimental Birth Sl. Calf No group (Kg) Weight (Kg) 30 days (Kg) 60 days (Kg) 90 days (Kg) 120 days (Kg) No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

D173 D175 D176 D185 D188 D191 D174 D177 D178 D183 D186 D192 D181 D182 D184 D189 D190 D193

25% Extra 25% Extra 25% Extra 25% Extra 25% Extra 25% Extra 50% Extra 50% Extra 50% Extra 50% Extra 50% Extra 50% Extra Control Control Control Control Control Control

29 26 22 19 20 20 25 21 17 26 26 25 24 23 22 31 25 23

35 34 33 27 25 24 35 32 26 35 30 29 31 28 28 38 33 26

45 40 39 39 39 32 43 41 37 47 45 35 38 37 37 43 43 39

57 54 56 51 54 39 56 55 52 60 59 52 47 46 47 55 51 46

71 70 80 64 64 56 70 78 76 72 77 54 55 60 54 61 56 55

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feed restricted quantities of milk to calves because of cost and the perception that increased milk intake may lead to a higher incidence of diarrhoea, reduced calf starter feed intake and reduced mass gain. Results from several studies indicate that feeding more milk or high-quality milk replacer does not cause diarrhoea ( Jasper et.al., 2002).

STATISTICAL ANALYSIS The overall result indicated that there was a significant increase (P<0.05) in daily average weight gain in animals fed with 25 percent and 50 percent extra milk when compared to animals in the control group. It was also observed that the daily average weight gains recorded in animals in the second and third group were significantly higher (P<0.05) than that of the control during the fourth month of age indicative of the carry over effect of the feeding system.

DISCUSSION Accelerated milk feeding adds economy by reducing calf mortality rate and cutting calving interval (Terre et al., 2006). Most dairy producers

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The concept of accelerated feeding for young milk-fed calves is now well-accepted as an alternative to traditional restricted feeding. Research and field experience have highlighted many important aspects that are required for successful implementation of accelerated milk feeding systems. A step-down or gradual weaning process facilitates a smoother transition to dry feed ( Flower and Weary, 2001). Colostrum-deprived calves or calves that are undergoing transport stress will not respond as well to increased amounts of milk and may in fact be impacted negatively. Benefits to accelerated milk-feeding programs include: decreased age at first calving , improvements in health, and increased milk production ( Kung et.al., 1997). Adequate milk feeding during younger ages decreases stress in calves and thus improves the performance (Fiems et.al., 1982, Diaz et.al., 2001). Based on the result of this study, it is recommended that a higher level of milk feeding regime during pre-weaning state will not only influence the growth rate during the period but also beneficially contribute to the growth performance in later stages. Ongoing research will provide the necessary input variables to model the overall economic impact of accelerated milk feeding programs

REFERENCES Davis, R., VandeHaar, C. A., Wolf, J. S., Liesman, L. T., Chapin and Weber, M.S. 2011. Effect of intensified feeding of heifer calves on growth, pubertal age, calving age, milk yield, and economics. J. Dairy Sci. 94:3554-3567. Diaz, M. C., Van, A. M. E., Smith, J. M., Kelsey, J. M and Hutten, E. L .2001. Composition of growth of Holstein calves fed milk replacer from birth to 105 kilogram body weight. J. Dairy Sci. 84: 830-842

different amounts of milk on performance, health, and absorption capability of baby calves. J. Dairy Sci. 67: 2957-2963 Kung, L. J., Demarco, S., Siebenson, L.N., Joyner, E., Haenlein, G.F.W. and Morris, R.M. 1997.An evaluation of two management systems for rearing calves fed milk replacer. J. Dairy Sci., 80: 2529-2533 Pettersson, K., 2000. Results from a questionnaire: housing, feeding and management of dairy calves in Sweden. In: Tielen, M.J.M., Voets, M.T. (Eds.), Proceedings of the 10th International Congress on Animal Hygiene, vol. 1. Maastricht, The Netherlands, pp. 421-425.

Fiems, L.O., Boucque, C.V., Cottyn, B.G and Buysse, F.X., 1982. Effect of feeding techniques and age at weaning on the performances of bucket-fed and suckling reared calves. In: Signoret, J.P. (Ed.), Welfare and Husbandry of Calves, Current Topics in Veterinary Medicine and Animal Science, vol. 19, pp. 149-167.

Terre,M. Bach, A., Geng, D and Devant, M. 2006. Performance and behaviour of calves reared in groups or individually following an enhanced-growth feeding programme. J. Dairy Res. 73: 480-486

Flower, F. and Weary, D.M .2001. Effects of early separation on the dairy cow and calf. Separation at 1 day and 2 weeks after birth. Appl. Anim. Behav. Sci. 70:275-284.

Tozer, P. R and Heinrichs, A .J. 2001 .What affects the costs of raising replacement dairy heifers: A multiple-component analysis. J. Dairy Sci. 84: 1836-1844

Jasper, J. and D. M. Weary. 2002. Effects of Ad Libitum Milk Intake on Dairy Calves. J. Dairy Sci. 85 :3054-3058.

Von , A. G., Wolf, F. Hรถtzel, M and Weary, D. M.. 2006 .Effects of continuous versus periodic milk availability on behavior and performance of dairy calves. J. Dairy Sci. 89: 2126-2131

Huber, J. T., Silva ,A. G., Campos, O. F and Mathieu, C. M. 1984. Influence of feeding

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RESEARCH ARTICLE

MORPHOGENESIS OF MEDULLA OBLONGATA IN GOAT FOETUSES* K. M. Lucy1, K. R. Harshan2, J. J. Chungath3 and N. Ashok4 College of Veterinary and Animal Sciences, Mannuthy. *

Part of the PhD thesis submitted by the first author to Kerala Agricultural University.

ABSTRACT Prenatal morphogenesis of medulla oblongata (MO) was studied using 46 goat foetuses ranging from 2.5 cm CRL (40 days of gestation) to 41.5 cm CRL (full term). By 40 days of gestation, the roof plate region of the rhombencephalon expanded enormously and as a result, the entire alar and basal plates of the neural tube were displaced laterally and ventrally. Nuclei first appeared in medulla by 48 days (4.0 cm CRL) and nerve fibres crossing in different directions broke up the gray substance into a mixture of gray and white mater, the reticular formation. Trapezoid body started developing by 48 days. Medullary pyramids appeared on the ventral surface by 81 days of age (13.0 cm CRL). Percentage contribution of medulla oblongata to the total brainstem weight increased progressively during gestation (from 13.90 percent in second month to 17.57 percent in the fifth month). When compared to cerebrum and cerebellum, the MO along with the other regions of the brainstem was noted to be a slow growing region. During second month, contribution of ¹Associate Professor, CV&AS, Mannuthy, ²Professor (Retired), ³Professor and Head, CV&AS, Pookot, 4Professor and Head, Department of Veterinary Anatomy and Histology, CV&AS, Mannuthy. <28 < J. Ind. Vet. Assoc., Kerala. 10 (3)

MO to the total brain weight was 4.69 percent, which gradually reduced to 3.46 percent in the fifth month indicating rapid growth of the region in the first half of gestation followed by a gradual “cephalic shift” of function from phylogenetically older brainstem to the higher cerebral and cerebellar cortices. Towards term, this region was well developed and the relative maturity of the MO in goats at birth justifies the classification of goat as a prenatal brain developer. Key words: Medulla oblongata, Prenatal development, Goat.

INTRODUCTION Medulla oblongata is the caudal portion of the brain, located between the pons rostrally and spinal cord caudally, resting on the basioccipital bone. Postnatal studies on the histomorphology of the hindbrain have been made in different domestic animals (Jenkins, 1978; King, 1987; Rizzo, 2006 and Konig and Liebich, 2007). However, prenatal developmental changes have not been well documented in ruminants. Hence, this study was planned to investigate prenatal morphogenesis of medulla oblongata in goats.

MATERIALS AND METHODS Prenatal morphogenesis of medulla oblongata (MO) in goats was studied using 46 goat foetuses ranging from 2.5 cm CRL (40 days of gestation) to 41.5 cm CRL (full term). The material available in the Department of Anatomy and those collected from the farms and clinics were used for the study. Body weight, body parameters and skull parameters of the subjects were recorded. The age of the foetuses was calculated from the 0.33

formula, W = 0.096 (t-30) derived by Singh et.al. (1979) for the goat foetuses, where 'W' is the body weight of the foetus in g and 't' is the age of the foetus in days. Based on the age, the foetuses were divided into four groups representing second, third, fourth and fifth months of gestation. The heads were separated at the occipito-atlantal junction and the brain was then carefully dissected out and fixed in 10 percent neutral buffered formalin. After recording the whole brain parameters, the MO was separated at the caudal border of the pons (rostral boundary) and the rostral limit of origin of first pair of cervical spinal nerves (caudal boundary). Measurements were taken and the data were analysed statistically (Snedecor and Cochran, 1985). Standard procedures were adopted for histoarchitectural studies (Luna, 1968).

RESULTS AND DISCUSSION Development in the Second Month Medulla oblongata, the caudal most segment of the brainstem, extended from the level of first pair of cervical spinal nerves to the caudal edge of the pons (Fig. 1). It lay on the unossified basioccipital and this cartilaginous skeleton developed in the sixth week of gestation.

Measurements of medulla oblongata at different stages of gestation are given in table 1. By 40 days, the roof plate region of the embryonic rhombencephalon expanded enormously. As a result, the entire alar and basal plates of the neural tube were displaced laterally and ventrally. Arey (1957) compared this to an opened book whose hinge was the floor plate. The lumen became the fourth ventricle covered dorsally by the thin, single layer of ependyma, the roof plate. This constituted the anterior and posterior medullary vela (Figs. 2 and 3). These vela were continuous with the cerebellum cranially and the roof of the central canal of spinal cord caudally. The sulcus limitans present on the ventrolateral wall of the fourth ventricle provided the plane of division of the medulla into a ventromedial basal plate and a dorsolateral alar plate. Similar observations were made in dog foetuses by Jenkins (1978). The lumen was filled with CSF. Nuclei appeared by 48 days and nerve fibres crossing in different directions broke up the gray substance into a mixture of gray and white known as the reticular formation. The trapezoid body started developing at 48 days (4 cm CRL). The point of emergence of the facial nerve from the medulla is illustrated in figure 4. The endolymphatic duct also could be seen within the petrous temporal. Vascular mesenchyme occupied the ependymal roof; the combined membrane, the tela choroidea, infolded as vascular tufts into the cavity of the myelencephalon constituting the choroid plexus of the fourth ventricle. This developed in the goat foetus in the sixth week of gestation. Keith (1947) reported that the choroid villi developed on the ventricular surface of the caudal medullary velum at eight weeks in the human foetus and CSF was being produced during Vol. 10 Issue 3 December 2012 JIVA <29 <

the third month. Later during the seventh week, the ependymal roof plate became broad and thin. The cavity of the rhombencephalon thus expanded to the sides, flattened dorsoventrally and was filled with CSF. The rhombic lip, ridge where the tela joined the alar plate was made up of three to four layers of cells. Harrison (1978) reported that the cells of rhombic lip were actively mitotic and provided large number of neuroblasts, which migrated cephalad into the ventral aspect of the hindbrain where they formed the pontine nuclei and the olivary nuclear complex. Caudally the medulla oblongata was continuous with the spinal cord. The fourth ventricle narrowed posteriorly to be continued as the central canal of spinal cord. Dorsal wall of this region showed thickened epithelium constituting the circumventricular organ. Medulla oblongata contributed 4.69 percent of the brain weight and 13.90 percent of the brainstem weight at this stage.

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Fig. 1 Dorsal surface of brain (55 days) 1. Cerebrum; 2. Corpora quadrigemina; 3. Cerebellum; 4. Medulla oblongata

Fig. 2 C.S. of the medulla oblongata at the level of rostral medullary velum (48 days). H&E. x 100 1. Medulla oblongata; 2. Nuclear aggregation; 3. Fourth ventricle with CSF; 4. Rostral medullary velum; 5. Body wall; 6. Internal glial limiting membrane

RESEARCH ARTICLE Development In The Third Month

Fig. 3 C.S. of the medulla oblongata at the level of caudal medullary velum (48 days). H&E. x 100 1. Caudal medullary velum; 2. Choroid plexus; 3. Fourth ventricle with CSF; 4. Medulla oblongata; 5. Nuclear aggregation; 6. Median sulcus; 7. Sulcus limitans

Fig. 4 C.S. of the medulla oblongata showing the facial nerve and endolymph duct (48 days). H&E.x100 1. Medulla oblongata; 2. Facial nerve; 3. Endolymph duct; 4. Petrous temporal bone

Fig. 5 C.S. of the medulla oblongata showing foramen of Lushcka and caudal medullary velum. (76 days). H&E. x 100 1. Foramen of Lushcka; 2. Caudal medullary velum; 3. Choroid plexus; 4. Sulcus limitans

The medullary pyramids could be distinguished from 81 days (13 cm CRL) and were in the form of longitudinal ridges on either side of the ventral median fissure but they were not widely separated in the rostral portion. These agree with the findings of Dellmann and Mc Clure (1975) in small ruminants. However, in cattle the pyramids were widely separated at the point of emergence from the caudal aspect of pons. The dorsal surface of the medulla formed the floor of fourth ventricle as noted in the second month. The caudal medullary velum projected from the dorso-medial angle of medulla oblongata. The fourth ventricle communicated with the subarachnoid space by the foramen of Lushcka (Fig. 5). Floor of the ventricle was marked by a deep median sulcus, which became shallower rostrally. On each side of the median sulcus was a continuous ridge, the medial eminence, bounded laterally by the sulcus limitans as observed by Truex and Carpenter (1969) in man and Dyce et al. (1996) in domestic animals. The medial eminence, or trigonum hypoglossi was formed by the nucleus of hypoglossal nerve. The lateral eminence was occupied by the caudal poles of the medial and inferior vestibular nuclei. In between the medial and lateral eminences was the intermediate eminence, the trigonum vagi. Mean weight of medulla oblongata was 0.220 ± 0.032g during third month. Width of medulla oblongata was more than its height throughout gestation. Average length and width of medullary pyramids were 0.805 ± 0.031cm and 0.230 ± 0.013cm, respectively. The trapezoid body was clearly demarcated and the mean rostrocaudal distance was 0.117 ± 0.005cm. Vol. 10 Issue 3 December 2012 JIVA <31 <

Dellmann and Mc Clure (1975) reported that the trapezoid body was more clearly demarcated in small ruminants than in cattle.

Development In The Fourth Month Medulla oblongata contributed 17.57 percent of the brainstem weight. Percentage contribution of medulla oblongata to the total brainstem weight increased progressively during gestation. During the fourth month, mean length, width and thickness of the medulla oblongata were 1.183 ± 0.027cm, 0.855 ± 0.019cm and 0.682 ± 0.011cm, respectively. Unlike in the pons region, maximum width of medulla oblongata exceeded the maximum height. Morphological features did not change much during the fourth month. Length and width of medullary pyramids increased 15.03 and 93.04 percent, respectively from third to fourth month. More increase in width corresponded to growth of cerebrum since these fibres have their origin in the cerebral cortex.

Development In The Fifth Month Trapezoid body was clearly demarcated from the pons. Cranio-caudal length of trapezoid body was 0.298 ± 0.003cm. Mean weight of medulla oblongata increased three-fold during fifth month. Corresponding changes were also noticed in the length, height and width of medulla oblongata (Table. 1). During second month contribution of medulla oblongata to the total brain weight was 4.69 percent which gradually reduced to 3.46 percent in the fifth month indicating rapid growth of the region in the first half of gestation followed by a gradual “cephalic shift” of function from phylogenetically older brainstem <32 < J. Ind. Vet. Assoc., Kerala. 10 (3)

to the higher cerebral and cerebellar cortices. When compared to other divisions of brain, the medulla oblongata along with other regions of brainstem was noted to be a slow growing region. Ventral median fissure was flanked by the pyramids. Mean length and width of medullary pyramids were 1.585 ± 0.088 cm and 0.656 ± 0.008 cm, respectively. Grossly, medulla oblongata was adult-like during the fifth month (Fig. 6).

Fig. 6 Dorsal view of brainstem (124 days) 1. Thalamus; 2. Pineal gland; 3. Rostral colliculus; 4. Caudal colliculus; 5. Fourth ventricle; 6. Medulla oblongata

The medulla oblongata is a great suprasegmental conveyor and co-ordinator for pathways and nuclei involved with vital regulatory and protective processes which affect the whole body. Towards term, this region was well developed and the relative maturity of the MO in goats at birth justifies the classification of goat as a prenatal brain developer. Foetal brain is most vulnerable when it is growing rapidly and nutritional deficiencies and diseases during the growing period can cause permanent damage.

REFERENCES Arey, L.B. 1957. Developmental Anatomy. 6th edn, W.B.Saunders Company, Philadelphia. pp. 454-501.

Dellmann, H.D and Mc Clure, R.G. 1975. Central nervous system. Sisson and Grossman's The Anatomy of the Domestic Animals. 5th edn, (Ed.) Getty R. W.B. Saunders Company, Philadelphia. pp.1065-80. Dyce, K.M., Sack, W.O. and Wensing, C.J.G. 1996. Textbook of Veterinary Anatomy. nd 2 edn, W.B. Saunders Company, Philadelphia. pp. 259-324.

Konig, H.E. and Liebich, H.G. 2007. Veterinary Anatomy of Domestic Animals, Text Book and Colour Atlas. 3rd edn, Schattauer, New York. pp: 40-48. Luna, L.G. 1968. Manual of Histological Staining Methods of the Armed Forces rd Institute of Pathology. 3 edn, Mc GrawHill Book Company, New York. p.258.

Harrison, R.G. 1978. Clinical Embryology. Academic Press, London. p. 250.

Rizzo, D.C. 2006. Fundamentals of Anatomy and Physiology. 2nd edn, Thomson Delmar Learning, Australia. pp: 244-69.

Jenkins, T.W. 1978. Functional Mammalian Neuroanatomy. 2nd edn, Lea and Febiger, Philadelphia. p. 480.

Singh, Y., Sharma, D.N. and Dhingra, L.D. 1979. Morphogenesis of the testis in goat. Indian J. Anim. Sci. 49: 925-31.

Keith, A. 1947. Human Embryology and th Morphology. 6 edn, Williams and Wilkins, Baltimore. pp: 117-53.

Snedecor, G.W. and Cochran, W.G. 1985. Statistical Methods. 7th edn, The Iowa State University Press, USA. p. 313.

King, A.S. 1987. Physiological and Clinical Anatomy of the Domestic Mammals. Oxford University Press, New York. p.325.

Truex, R.C. and Carpenter, M.B. 1969. Human th Neuroanatomy. 6 edn, The Williams and Wilkins, Baltimore. p. 673.

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RESEARCH ARTICLE

ROLE OF FECUNDITY GENES IN PROLIFICACY OF SMALL RUMINANTS Asha Abraham¹ and Naicy Thomas² College of Veterinary and Animal Sciences, Mannuthy

INTRODUCTION The aim of every breeder is to get maximum profit possible from the animal he is rearing. This can be achieved by improving the genetic worth of the stock by proper selection methods. For the traits that are to be selected, their relative economic value should be established. Kidding/lambing percentage is the most important factor affecting profitability in small ruminants. Increasing prolificacy offers greater potential for improving reproduction rate and production efficiency. Only way to increase the numerical productivity (no. of kids produced per goat per year) is that those which are closely linked to reproductive parameters. Improvement of reproductive traits in livestock species has become of increasing interest,especially in small ruminants, where small increases in litter size can equal large gains in profit.

obtain selection improvements in traits associated with reproduction, since kidding/lambing percentage is lowly heritable. Differences among does/ewes in litter size (single, twin, triplet) are largely due to non-genetic factors, such as management and nutrition. Genetic change is permanent but, nutrition and management vary from year to year. Common strategy for increasing prolificacy via genetic means is to select ewes that are more likely to produce multiple births and to select rams that are more likely to sire prolific daughters. This can be accomplished through simple selection based on birth type or by selecting for a composite trait, such as weight of young ones weaned. In the absence of any other information, it is best to select males and replacement female lambs/kids that are born as multiples from young females.

FECUNDITY GENES PROLIFICACY

1. Booroola gene (FecB)

Prolificacy is measured as the ability of a female animal to produce large number of young ones in their life span through high ovulation rate and high embryo survival. But, it is difficult to

It is a single gene in chromosome 6 in sheep which is the main reason for higher prolificacy of certain breeds. This gene has effects on granulosa cell maturation, oocyte development and its function. The increase in prolificacy is due to autosomal mutation that occurred in this gene which causes increase in ovulation rate and litter

¹MVSc scholar, ² Assistant Professor, Centre for Advanced Studies in Animal Genetics and Breeding, College of Veterinary and Animal Sciences, Mannuthy <34 < J. Ind. Vet. Assoc., Kerala. 10 (3)

Feeding maintenance quality hay or pasture is enough for lambing rates of 240-300 percent in heterozygous Booroola ewes. Ancestor and original source of Booroola Merino sheep FecB gene is the Garole Sheep from the Sunderban (Bengal) area of India. Garole is a small ewe of 12-15 kg, with a mean litter size of 2.3 lambs, adapted to hot, humid, swampy conditions of rice paddies. 10 ewes and 2 rams were originally imported to Australia in 1792, from Calcutta, India. The FecB gene mutation is found in the Garole sheep. Genetic evidence supports the historical records that the prolific Garole sheep, when introduced to Australia, bred with the much larger 35-40 kg Merino sheep. Garole sheep are virtually all homozygous for the FecB gene (BB).

2. GDF9 / FecG gene

Normal and FecB gene carriers kept at Marshall Building in Edinburgh size. Term â&#x20AC;&#x153;Booroolaâ&#x20AC;? was taken from the name of the ranch in Australia, where the sheep carrying single gene for prolificacy were first discovered. Booroola gene can be transferred to any other breed by crossbreeding. This FecB gene is responsible for the higher prolificacy of Finn sheep, Romanov, Booroola Merino and British Milk Sheep. Up until now, the most numerous breed of sheep containing the FecB gene is Booroola Merino, the breed it was originally discovered in.

Growth differentiation factor 9 (GDF9) is seen in chromosome 5 in sheep and goat. They are expressed in oocytes and play an important role in ovarian folliculogenesis. Single strand conformation polymorphism (SSCP) studies of exon 1 and flanking of GDF9 gene reveals two silent mutations (183A>C and 336C>T) in AA genotype in comparison to BB genotype. Studies show that allele A has correlation with prolificacy in Jining Grey goat. (Chu, 2004).

3. BMP15 / FecX gene Bone morphogenetic protein 15 (BMP15) is an X linked gene seen in sheep and goat. BMP15 gene is essential for oocyte and follicular development. Higher prolificacy of Inverdale, Lacaune, Belclare, Small Tailed Han ewes and Jining Grey goats is due to this gene. Single strand Vol. 10 Issue 3 December 2012 JIVA <35 <

conformation polymorphism (SSCP) studies show SNPs in exon 1 and exon 2. Two point mutations (G963A and G1050C) are found in AB genotype in comparison to AA genotype (Vacca, 2010).

4. POU1F1 gene POU domain, class 1, transcription factor 1 (POU1F1) is otherwise known as Pit1 and GH factor 1. It is seen in chromosome 3 in sheep. POU1F1 is an important transcription factor for Growth Hormone. Single strand conformation polymorphism studies reveals six mutations C256T in exon 3, C53T and T123G in intron, G682T, T723G and C837T in exon (Feng, 2012).

and 220T"C) in AB genotype in comparison to AA genotype. (Zhang, 2007).

7. FSH receptor gene FSH receptor gene is seen in Chromosome 2 in sheep. It affects the follicular growth and hence plays an important role in prolificacy. Variations in aminoacid sequence of receptor protein are due to point mutations. Single strand conformation polymorphism studies shows that polymorphisms at codon 307 and 680 influence responsiveness to FSH and affects prolificacy (Tisdall, 1995).

8. KiSS-1 gene 5. Estrogen Receptor gene Estrogen receptor gene in sheep has two regions - ESR1 in chromosome 6 and ESR2 in chromosome 14. Single strand conformation polymorphism studies show SNP of exon 1 of Estrogen Receptor (ESR) gene. AC"G mutation was noticed at the 363bp of exon 1 in BB genotype on comparison to AA genotype. Marked assisted selection (MAS) of animals with estrogen receptor gene polymorphisms can be done to increase litter size and thus increase in economic value to mutton producers (Xiao-Dan, 2005).

6. Prolactin receptor gene Prolactin receptor gene is seen in chromosome 5 of sheep. This gene mainly interacts with prolactin and thus increases prolificacy. Single strand conformation polymorphism studies of exon 10 of prolactin receptor gene reveals two mutations (186G"A

Kisspeptin (KiSS-1) gene is important for proper GnRH function and thus affects prolificacy. It is seen in chromosome 1 in sheep. Single strand conformation polymorphism studies reveals that polymorphism of intron 2 results in two SNPs T2643C and 8bp base deletions (2677AGTT CCCC) giving rise to four different genotypes CC, TT, TG and TC (Hou, 2011).

9. INH gene INH gene (Inhibin gene) is essential for normal oocyte and follicular maturation and affects prolificacy. Inhibin genes - INHA and INH창A are found in chromosome 2 in sheep. Single strand conformation polymorphism studies reveal a C865T silent mutation in exon of INHA gene (Yuanqing He, 2010).

CONCLUSION Genetic improvement of reproductive

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traits has traditionally been restricted to use of quantitative genetic methods but gain has been limited when using these methods. Recent improvement in molecular genetics provided that the major genes associated with reproduction can be utilized in breeding through marker-assisted selection (MAS). Reproductive traits are often suggested as prime targets for marker-assisted selection for their low heritability and the fact that the trait can be measured only in one sex. The fecundity genes has posed the unique and exciting opportunity to add a high level of prolificacy to sheep and goat, that fit the environment well, without having to add undesirable traits of other breed. These genes have proved to be trackable and persistent, after its introduction to a breed. Fecundity traits offer a new option that can allow certain breeds producing lambs/kids to obtain a high level of prolificacy by genetic introgression of desired allele of the fecundity gene. The presence of fecundity genes, Booroola and Inverdale has been proved as a cause for high prolificacy for Malabari goats also. So, by using genetic molecular markers we can detect the mutations which results in high prolificacy in certain breeds (Davis, 2004). Breeders can select males from these breeds to incorporate into crossbreeding and can attain high level prolificacy. Speed and efficacy of selection is expected to increase by the use of these molecular markers in selection.

Davis, G. H. 2004. Fecundity genes in sheep. Anim. Reprod. Sci. 82-83:247-253 Feng, T., Chu, M. X., Cao, G. L., Tang, Q. Q., Di, R., Fang, L., and Li, N. 2012. Polymorphisms of caprinePOU1F1 gene and their association with litter size in Jining Grey goats. Mol. Biol. Rep. 39(4):4029-4038 Hou, J. X., An, X. P., Wang, J. G., Song, Y. X., Cui, Y. H., Wang, Y. F., Chen, Q. J. and Cao, B. Y. 2011. New genetic polymorphisms of KiSS1gene and their association with litter size in goats. Small Ruminant Research. 96 (23): 106-110. Tisdall, D. J., Watanabe, K., Hudson, N. L., Smith, P. and Mc Natty, K. P. 1995. FSH receptor gene expression during ovarian follicle development in sheep. J. Mol. Endocrinol. 15: 273-281. Vacca, G. M., Dhaouadia, A., Rekikb, M., Carcangiua, V., Pazzolaa, M. and Dettoria, M. L. 2010. Prolificacy genotypes at BMPR1B, BMP15 and GDF9 genes in North African sheep breeds. Small Ruminant Research. 88(1): 67-71. Xiao-Dan, B. I., Chu Ming-Xing, Jin Hai-Guo, Fang Li and Ye Su-Cheng. 2005. Estrogen Receptor as a Candidate Gene for Prolificacy of Small Tail Han Sheep. Acta Genetica Sinica. 2005 -2010

REFERENCES

Yuanqing He, Xiaoke Ma, Xiaoyong Liu, Cunxia Zhang and Jun Li. 2010. Candidate gene polymorphism and its association in Chinese goats. J. Agri. Sci. 2:1.

Chu, M. X., Li, B. X., Wang, J. Y., Ye, S. C and Fang, L. 2004. Association between PCR-SSCP of growth differentiation factor 9 gene and high prolificacy in Small Tail Han sheep. Anim. Biotechnol. 15(2):111- 120

Zhang, G. X., Chu, M. X., Wang, J., Fang, L and Ye, S. C. 2007. Polymorphism of exon 10 of prolactin receptor gene and its relationship with prolificacy of Jining Grey goats. Yi Chuan. 29(3):329-336 Vol. 10 Issue 3 December 2012 JIVA <37 <

RESEARCH ARTICLE

PARTICIPATION OF SELF HELP GROUP MEMBERS AND NON- MEMBERS IN PANCHAYATI RAJ SYSTEM A COMPARATIVE STUDY* 1

Anu George , P. J. Rajkamal and R.S. Jiji

College of Veterinary and Animal Sciences, Mannuthy. *

Part of M.V.Sc thesis submitted by the first author to Kerala Agricultural University

ABSTRACT Level of participation of livestock farmers in Panchayati Raj system of democratic decentralisation was analysed in the study. A comparison was made between Self Help Group (SHG) members and non members in terms of level of participation. The results showed that the level of participation was medium to high for SHG members, whereas, it was low to medium for most of the non members. The difference was statistically significant. Key words: Panchayati Raj, Democratic decentralisation, Self Help Group, Participation

INTRODUCTION Democratic decentralisation, often referred as Panchayati Raj system in India, is meant for the transfer of authority, responsibility and accountability from central to local governments. Any rural development programme in order to be effective should ensure people's liberal participation. Thus, in the Panchayati Raj system of local self governance, the emphasis has shifted towards the participation of people through 1

PhD Scholar, 2Professor and Head and 3Associate Professor Department of Veterinary and Animal Husbandry Extension College of Veterinary and Animal Sciences, Mannuthy, Kerala <42 < J. Ind. Vet. Assoc., Kerala. 10 (3)

Panchayati Raj Institutions (PRIs), in planning, formulation and execution of development programmes. Decentralisation is effected in terms of political, financial and administrative dimensions. Panchayati Raj system of democratic decentralisation was revamped in Kerala state since 1995. It has coordinated grass root level planning. It is vested with the authority of formulation and implementation of projects. Local needs are found out through the gram sabha and resources are allocated based on the priority of needs. People participate in the governance, decision making and implementation. Transparency and accountability are the key features of the system. Local self governance is important because, weaker sections of the society have greater chances of participation in decision making which might affect their lives directly or indirectly. The common people, especially the livestock farmers, have vested much hope in Panchayati Raj believing that need-based participatory planning at grass roots can take place to their own benefit. They are known to participate in the Panchayati Raj system at varying levels hopeful of receiving benefits. Self Help Groups (SHGs), neighbourhood groups (NHGs), Gramsabha etc. are the venues of participation.

Self Help Group (SHG) is a small group of rural people, usually 10-20 in number, gathered voluntarily to undertake some common income generating activities through mutual trust and mutual help (Arunkumar, 2005). In Kerala, SHGs are gaining more impetus nowadays and many are taking livestock rearing as the key activity. Furthermore, the poverty eradication mission of Kerala Government, Kudumbasree is also organizing women SHGs including that of livestock based SHGs. The present study was carried out with the objective of comparing the level of participation of SHG members and non members in PRIs.

METHODOLOGY The study was conducted in Thrissur district of Kerala state. Two block panchayats, viz., Ollukkara and Irinjalakuda were selected purposively, since one of the key activities of SHGs in these blocks was livestock rearing. There were 65 viable livestock- based SHGs in these blocks. The list of SHG members was prepared with the help of office bearers of these SHGs. One hundred members were selected from among the total of 675 members, and they formed the sample of SHG members. The sample of non-members comprised of 100 livestock owners selected randomly from the list prepared with the help of secretaries of milk co-operative societies and extension personnel working in these blocks. Thus a total of 200 livestock farmers formed the sample of the study. The level of participation in PRIs meant the degree of respondents' participation in the various Panchayati Raj bodies and activities concerned with the planning and implementation of projects. It was measured by personal interview method

using the schedule developed for the purpose. The findings were expressed as percentage. Based on the Delenius-Hodges cumulative f method, the respondents were grouped into three groups as low, medium and high for level of participation.

RESULTS AND DISCUSSION Panchayati Raj system of participatory planning replaced the earlier Community Development Programme to ensure people's participation in community development. According to Mishra (1994) participation means co-operating or taking part in something and the mere presence, even the silent presence of an individual or a representative of an organisation at different levels can be taken as participation. Level of participation in PRIs Table. 1. Distribution of respondents based on the level of participation in PRIs n=200 Frequency (Percentage)

Sl. Category No

Score

Low

Medium

8 - 31

High

> 31

100

Total

SHG Nonmembers members

Data in table 1 show that majority (79 percent) of the SHG members were having high level of participation in PRIs. For the rest 21 per cent, the level was medium. There was nobody in the low category. But the situation is different as far as non members are concerned. The level of participation in PRIs was low for 60 percent of the non SHG members, medium for 38 percent and high for only 2 percent of the respondents. As for Vol. 10 Issue 3 December 2012 JIVA <43 <

non SHG member livestock owners, reporting either a medium level or low key participation in PRIs, was not at all the result normally expected. This is a precarious situation and as such this system does not seem to give any hope as far as livestock owners are concerned. Remedial measures are urgently needed on a war footing before it collapses. Z test on selected variables Table 2. Z test on selected variables Mean ± SE Sl. Variables SHG NonZ value No members members

CONCLUSION Majority of livestock owners were reporting either a medium level or low key participation, where as majority of SHG members level of participation in PRIs was reported to be high which was statistically significant. Capacity building through appropriate training programmes would be the best strategy for better participation. The results indicate the need to reconsider the nature and intensity of SHG promotion. SHG promotion cannot be considered as a one-shot, simple input. It has to be more strategic, adaptive and long-term.

1 Level of participation 28.41±1.56 22.96±2.79 17.02** ** (p<0.01)

Z test indicated that (table 2) there is significant difference between the SHG members and non members in terms of level of participation in Panchayati Raj. While studying the perception of veterinary surgeons of Thrissur District of Kerala about people's participation in PRIs, Tajne (2003) confirmed that majority of them felt the extent of people's participation to be only somewhat satisfactory. David (1998) also reported that participation of people in the planning process to be not satisfactory. There are apparent synergies between SHGs and local politics also. Through membership in SHGs rural people can gain experience in regular meetings, taking decisions and allocating money. Since SHG members have more public contact than others, they can effectively participate in campaigning too (Anon, 2006). SHG membership generally contributes to women getting nominated in election to local bodies. <44 < J. Ind. Vet. Assoc., Kerala. 10 (3)

REFERENCES Anon. 2006. Self Help Groups in India-A study of the lights and shades. Executive summary of EDA Rural systems Pvt. Ltd in association with APMAS for CRS, USAID, CARE, GTZ/NABARD, p20. Arunkumar, D. 2005. A critical analysis of SwaShakti programme in Karnataka, M.Sc (Agri) Thesis, University of Agricultural Sciences, Dharwad, p112. David, R.P. 1998. Decentralised planning in Kerala-Case study of two panchayats. MA Project Report, Calicut University, p85. Tajne, S.B. 2003. Work environment of Veterinary Surgeons of Thrissur District for extension activities under Panchayati Raj. M.V.Sc. Thesis, Kerala Agricultural University, Thrissur, p124. Mishra, Y. 1994. People's participation in production process under watershed. Kurukshetra 42:28-30.

RESEARCH ARTICLE

THERMAL STRESS IN DAIRY CATTLE 1

A. Prasad , E. M. Muhammed , A. Kannan and T. V. Aravindakshan

College of Veterinary and Animal Sciences, Mannuthy, Thrissur

ABSTRACT Crosses of temperate cattle reared in tropical ambience lapse in heat tolerance, fertility and disease resistance. However, cross breeding zebu with high yielding exotic cattle seems necessary to meet the need for enhanced composite milk production per unit in view of food security. Thermal stress is considered as the main factor responsible for reduction of milk yield in tropical climate. Any model for the study of thermal stress in dairy cattle should encompass related effect of ambient parameters of temperature, humidity, wind speed and solar radiation as the stressor and behavioural, autonomic, neuroendocrine and immunological endpoints as responses. The responses reach different response states depending on the severity of the stressor. Response measurements are at behavioural, physiological, biochemical and cellular level, which also include hormone, protein and gene expression assays. By fitting these measurements to the described model we can work out the biological and economic cost of thermal stress and the level of adaptation of the 1

Assistant Professor, 3Associate Professor, Department of

Livestock Production Management, 2 PhD Scholar, 4 Professor, Department of Animal Breeding, Genetics and Biostatistics, College of Veterinary and Animal Sciences, Mannuthy, Thrissur.

dairy animal in question. Such studies taking in to consideration the diverse nature of climatic factors is imperative for finding ameliorative measures to reduce the thermal stress experienced by the existing cattle population and for the possible genetic and management strategies for evolving and maintaining a climatically adapted dairy stock in a state like Kerala. This review analyses suitable model for climatic adaptation studies in the hot and humid climate especially in small holder production systems.

INTRODUCTION Challenge before the scientific community of the tropical world is to find ways to enhance milk production in the prevailing climatic conditions. Historically the traditional livestock production largely depended on heat tolerant native breeds that produced less milk compared to temperate exotic breeds. The dairy sector now largely comprises of extensive and expanding crossbred population in Kerala. For crossbreds, increased air temperature, and humidity measured as Temperature Humidity Index (THI) above critical thresholds are related to low dry matter intake (DMI) and to reduced efficiency of milk production (West, 2003). Vol. 10 Issue 3 December 2012 JIVA <45 <

The Zone of Thermoneutrality (Fig. 1) with in which no additional energy above maintenance is expended to heat or cool the body (for livestock it is between -0.5 to 200C) and the upper critical temperature (B on the right side ) may reach to 250

26 C (West, 2003). The ambient temperature of hot humid region is above this critical temperature during several months of each year. Effective environmental temperature is a combined effect of ambient temperature and humidity. The combined effect is quantified as Temperature Humidity Index (THI). The normal THI to maintain production in dairy cattle is 72. In our state, during most days in a year it is hot and humid and hence the THI is high enough to cause significant heat stress.

Fig.1 Kleiber's law of metabolic heat production and core body temperature as influenced by environment temperature. C = lower critical temperature D= point of reduction of metabolic heat. B on the left side = lower point of zone of thermal neutrality below which chemical regulation is needed to maintain homeothermy. B on the right side = upper critical temperature.

So the dairy cattle especially those with exotic blood cannot express its optimum production potential. Constant exposure of cattle <46 < J. Ind. Vet. Assoc., Kerala. 10 (3)

to high temperature causes a rise in its rectal temperature, a decline in feed intake, increase in water intake, a decrease in production of milk, changes in milk composition, reduction in growth and even a loss in body weight. This is the reason for deterioration in the performance of temperate dairy cattle when introduced into tropical countries. (Sastry and Thomas, 2005). The extent of reduction in feed intake is proportional to the thermal stress i.e. how long and how much the cow is subjected to temperature beyond thermo neutral zone. VFA is reduced in summer, especially acetic acid. Feed efficiency is considerably reduced. Digestible energy o

utilization was 60 percent at 21 C but only 40 o

percent after 7 days exposure to 32 C and 31 percent after 14 days exposure (Jones and Stallings, 1999). High ambient temperature and humidity causes depression in yield of fat and SNF. Lactose percentage is also depressed when cows are exposed to high ambient temperature. Genetic potential of well adapted local breeds that can be maintained without special feed concentrates or preventive health care are being increasingly recognized in this context. The prevailing harsh climate and the anticipated future climatic changes are real threats to the sustainability of the sector. An exact understanding of the impact of thermal stress on dairy cattle and possible ameliorative measures are thus extremely important. Thermal stress alone does not act separately in reducing the productivity of the animal, but multiple stresses act on the animal when nutrition is compromised (Sejian et.al. 2012). Since, interpretation of the effect of multiple stressors are difficult, multiple responses of the thermal stress alone is discussed here.

'Stress' conceptual clarity

2. Autonomatic nervous system

The word 'stress' is perceived by different people differently. Even among animal scientists, different disciplines approach stress in different angles. Here an attempt is made to elucidate the word stress in the context of thermal stress in dairy cattle. Stress is a part of life. All life forms have evolved mechanism to cope with the stresses of their lives. Moberg and Mench (2000) defined stress as the biological response elicited when an individual perceives a threat to its homeostasis. The threat is the stressor. When stress response truly threatens the animal's well-being then the animal experience distress.

3. Neuro Endocrine

The stressor in our context is the effective environmental temperature represented by the temperature, solar radiation, humidity and air movement in the immediate vicinity of the dairy cow. Our challenge is to determine when their stress becomes distress and to determine how to measure stress and distress in animals.

4. Immune response Now let us attempt to evolve a model forthermal stress in dairy cattle which better clarifies our problem and will help in attempting to assess it scientifically.

(Behavioural, autonomic, neuro endocrine, immunological)

Evolving a model for assessing thermal stress in dairy cattle Moberg (2000) divided stress response in to three general stages 1. Recognition of a stressor 2. Biological defense against the stressor and 3. Consequence of the stress response. A stress response begins with the CNS perceiving a potential threat to homeostasis. Then the cow develops biological response or defense that consists of four general biological defense responses 1. Behavioural Vol. 10 Issue 3 December 2012 JIVA <47 <

Challenges in measuring thermal stress

Behavioural response

Scientists rely on a variety of endocrine, behavioural, autonomic nervous system and immunological end points to measure stress. Unfortunately, none of these measures has proved to be a litmus test for stress. Further, complicating is inter animal variability in stress response.The above model demonstrates that all the responses to the perceived stressor can reach any one of the response state depending the severity of the stressor and the vulnerability of the animal. The vulnerability depends upon the modifiers of the responses. Modifiers which shape animal's organization of its biological defenses are early experience, genetics and age. The responses are not mutually exclusive but need to be measured independently for better assessment. After assessing them independently measures related to each response must be compared with normal values to arrive at the respective response state. As any one single response cannot be considered to be exclusively due to stress, the non parametric approach of considering the mode of response states for the entire array of tests may be taken as the animal's thermal stress level. Different responses and possible measurements are explained below and illustrated in Fig. 2.

A cow seeking shade during direct solar exposure is an example. But in case of dairy cattle behavioural response options are limited by confinement. When the severity of the stressor increases, this response can manifest as altered behaviour, pre pathological behaviour and even pathological behaviour. But all the animals will not show same degree of alterations in behaviour to the same degree of stressor.

Fig. 2. Extrapolation of Fig 1 to depict the response states of the animal when subjected to effective temperature above the upper critical temperature. (The column 1 - altered, 2- pre pathological and. 3pathological response states. <48 < J. Ind. Vet. Assoc., Kerala. 10 (3)

Autonomic nervous system response During thermal stress the autonomic nervous system affects a diverse number of biological systems including the cardiovascular, gastro intestinal, exocrine glands and adrenal medulla. The results are altered heart rate, blood pressure and gastrointestinal activity. But stress activation of autonomic nervous system is of relatively short duration. According to the grade of the stressor and the animal's vulnerability each of the autonomic nervous system responses also differs. Neuro endocrine Hormones secreted from hypothalamic pituitary system have a broad long lasting effect on the changes induced by body stress. The secretions of pituitary hormones have been implicated in failed reproduction, altered metabolism, immune competence and behaviour. The hypothalamic pituitary- adrenal (HPA) axis has been primary neuroendocrine axis. However, the secretion of prolactin and somatotropin has proven to be equally sensitive to stress and has considerable economic importance. Likewise, thyroid stimulating hormone and gonadotropins (LH and FSH) are either directly or indirectly modulated by thermal stress.

Increased secretion of the adrenal glucocorticoids and cortisol has been found to be associated with stress and investigators frequently cite an increase in circulating cortisol as a proof of stress. Colborn et.al., (1991) found that stallions secreted similar amounts of cortisol whether the stallions were restrained, exercised or permitted to mate with a mare. Serum cortisol concentrations are often used to evaluate stress, but due to the marked variability, faecal corticosterone has been used to evaluate stress in cattle (Morrow et.al., 2002). Dairy cattle secrets cortisol during restrained and when they are approached by strangers. So it is difficult to simply use secretion of cortisol or any other hormone to differentiate between non threatening stress and distress. Even among the same breed, some cows may produce more cortisol to the same amounts of thermal stressors. Immune response We have long attributed the increased incidence of disease in cows suffering from thermal stress. Immune system in its own right is one of the major defense systems responding to stressor (Dunn, 1988). Measurement of immune competence offers us a potentially powerful tool for evaluating the disease components of distress. According to Kumar et.al., (2011) heat stress is one of the wide varieties of factors which cause oxidative stress in-vivo. Reactive oxygen species (ROS), constantly generated in vivo as an integral part of metabolism cause oxidative stress when their level exceeds the threshold value. Superoxide ions, hydrogen peroxide and hydroxyl ions (ROS) produced as a result of heat stress are highly reactive and they cause damage to the polyunsaturated fatty acids of lipoprotein layer of cell membrane. This lipid peroxidation leads to the formation of lipid peroxides which are again

harmful to the cell components. The amount of lipid peroxides is measurable and hence can be used as reliable indicator of level of thermal stress. Reduced glutathione present in the animal acts as an anti oxidant by getting itself oxidized. So in a distressed animal the amount of reduced glutathione will be low. The heat shock response is a highly conserved cascade of altered protein and gene expression in animals. The altered intracellular proteins secretion is due to the concerted action of physiological stress response which constitutes a system wide gene network coordinated across a variety of cells and tissues to minimize effects of adverse environmental conditions. Endocrine and metabolic responses are as result of gene expression changes that include (1) activation of heat shock transcription factors (HSFs), (2) Increased expression of HSPs and deviated expression and synthesis of extra proteins, (3) Increased protein, glucose and amino acid oxidation and reduced fatty acid metabolism, (4) Endocrine system activation and (5) Immune system activation (Richter et.al., 2007). Diverse physiological stresses (thermodynamics, mutant proteins and oxidative injury) produce multiple changes in a cell that ultimately affect protein structures and function. Cells from different phyla initiate a cascade of events that engage essential proteins, the molecular chaperones, in decisions to repair or degrade damaged proteins as a defense strategy to ensure survival. Molecular chaperones such as the heat shock family of stress proteins (HSPs) actively participate in an array of cellular processes, including cytoprotection. The versatility of the ubiquitous HSP family is further enhanced by stress-inducible regulatory networks. Vol. 10 Issue 3 December 2012 JIVA <49 <

Biological cost of thermal stress Whether or not the stress altered functions are beneficial in helping the cow to cope is not our immediate concern. The changes in biological function during stress result in a shift of biological resources away from biological activities occurring before the stressor. For example, energy originally utilized for growth or reproduction might be needed by animal to cope with stress. This change in biological function during stress is the biological cost of stress. During prolonged stress or when stress is severe the biological cost is significant and the work of stress becomes a significant burden to the body. It is during such stress that the animal enters the next stages of stress pre pathology and pathology. In a hot humid climate like the one in Kerala, the dairy cows suffer from chronic distress which is the sum total of all the compensations or displacements caused by the thermal stress applied. This chronic distress represents the totality of discomfort, felt by the cow in a hot environment. The cows under this chronic distress will have reduced feed intake and increased water consumption. The body temperature will rise often stabilizing at a higher level. There will be shift in body water from intra cellular to extra cellular and extracellular to vascular space which will be mobilized to effect evaporative thermal cooling from skin and respiratory tract. Increased respiratory activity leads to excessive blowing out of carbon dioxide through the expired air thus upsetting carbonic acid bicarbonate buffer system. In order to maintain acid base balance, bicarbonate, mainly in the form of sodium bicarbonate or potassium bicarbonate is excreted through urine. Excessive loss of sodium <50 < J. Ind. Vet. Assoc., Kerala. 10 (3)

from body triggers corticosteroid hormone. Ultimately acid base and mineral balances are disturbed. Excessive moisture through evaporative channels can cause dehydration. Under hot condition, when subjected to water scarcity, energy utilization efficiency is reduced and reproductive efficiency is progressively affected. Acclimation and adaptability factor Acclimation is within life time phenotypic response to environmental stress and is a homeothermic process driven by endocrine system, whereas, adaptability involves evolutionary changes that occur over time scales covering multiple generations. Alterations in gene expression and changes in cellular signaling are key components of adaptability. The changes in gene expression mainly include that of HSPs. The major symptom in crossbred cattle is reduced feed intake, when the animals are under heat stress. Even if they are fed adlibitum, that too high quality feed, the animals do not take feed. This may be due to oxidative cellular stress that leads to cell starvation. Cell starvation often leads to inability of the cells to utilize glucose and other energy releasing molecules. When the cells are under starvation, growth, production and reproduction are compromised first, and then the vital functions of body. All these changes are associated with altered nutrient partitioning that accounts for production loss. In the acclimated state, metabolism is adjusted to minimize detrimental effects of increased thermal loads. We have to determine the basis of altered energy metabolism during thermal stress in exotic and indigenous animals which may lead to opportunities for improved animal performance via altered breeding strategies.

What contributed more to the natural selection are the fitness traits such as fertility and viability. The cattle breeds evolved in temperate and tropical climate are adapted to respective climate in terms of these fitness traits. When they are taken to alternate conditions fitness traits are maintained where as production traits are compromised. So it is not possible to get a high production from these breeds in alternate conditions (i.e. temperate breeds in tropical climate and tropical breeds in temperate climate).

CONCLUSION Thermal stress is a major threat to the viability and sustainability of milk production around the world, especially in tropical climate. The Inter-governmental Panel on Climatic Change (IPCC) has forecasted global warming leading to rise in average temperature by 1.8-40 C by the year 2100. Thus, the deteriorating prevailing condition is going to be aggravated unless measures to alleviate the thermal strain are addressed at appropriate level. For suggesting genetic, nutritional and management measures to overcome this challenge, scientific operational definition of thermal stress elucidated through suitable models are imperative. The model suggested in the present study is based on various reviews on different aspects of the subject. The model depicts the different levels of stress response to the perceived stressor and various methods to measure the responses. This model will also help in determining the level of acclimation and adaptability of cattle. REFERENCES Colborn, D. R., Thomson, D. L., Roth, T. L.,

Capehart, J. S. and White, K.L.1991. Response of cortisol and prolactin to sexual excitement and stress in stallions and geldings J. Anim. Sci.69:2556-2562. Hutcheson, D.P. and Cole, N.A. 1986. Management of transit stress syndrome in Cattle: Nutritional and Environmental Effects. J. Anim. Sci. 62:555-560. Jones, G.M. and Stallings, C.C. 1999. Reducing Heat Stress for Dairy Cattle. Virginia Cooperative Extension, Dairy Publication, 404-200. Kumar, S.B.V., Ajeet, K and Meena, K. 2011. Effect of heat stress in tropical livestock and different strategies for its amelioration. J. Stress Physiol. Biochem. 7 (1): 45-54. Moberg, G. P and Mench, J. A. 2000. The Biology of Animal Stress Basic Principles and Implications for Animal Welfare, CABI Publishing Morrow, C. J., Kolver, E. S., Verkerk, G. A. and Matthews,L.R. 2002. Faecal glucocorticoid metabolites as a measure of adrenal activity in dairy cattle. Gen. Comp. Endocrinol. 126: 229-241. Rao, G.S.L.H.V.P. 2003. Agricultural Meteorology Kerala agricultural University, Thrissur, Kerala, India, p.231. Richter, K., Haslbeck, M. and Buchner, J. 2010. The heat shock proteins: Life on the verge of death. Molecular cell. 40:253-266. Sastry, N. S. R and Thomas, C. K. 2005. Livestock th Production Management (4 ed.). Kalyani Publishers, New Delhi, pp. 13-15. Sejian, V., Naqvi, S. M. K., Ezeji, T., Lakritz, J and Lal, R. 2012. Environmental stress and amelioration in livestock production. Springer, Newyork . pp 129-144. West, J. W. 2003. Effects of heat stress on production in dairy cattle J. Dairy Sci. 86:2131-2144 Vol. 10 Issue 3 December 2012 JIVA <51 <

CLINICAL REPORT

SUCCESSFUL MANAGEMENT OF CHYLOTHORAX IN A DOG- A CASE REPORT G. Vijayakumar1, S. Sivaraman2, E. Venkatesakumar3 and M. Subramanian4 Veterinary College and Research Institute, Namakkal

INTRODUCTION Chylothorax is the accumulation of chyle in the pleural cavity and has been reported in dogs, cats, humans and other species (Ettinger and Feldman, 2010). Any disease or process that increases systemic venous pressures (i.e. right heart failure, mediastinal neoplasia, cranial venacava thrombi or granuloma) may cause chylothorax.

CASE HISTORY AND CLINICAL FINDINGS A non descript male dog aged about 8 years was brought to Veterinary College and Research Institute hospital with history of dyspnoea and not taking food for five days. Clinical examination of the dog revealed pale mucous membrane, cough, abducted elbows, severe dyspnoea, oral breathing / panting, lethargy, exercise intolerance and tachycardia. Auscultation revealed muffled heart sound and murmur. Haemato -biochemical examination did not reveal any abnormality. Thoracocentesis was done and about 520ml of chyle was removed on the day of presentation (Fig 1). Analysis of the pleural effusion revealed chyle (Fig 2) white coloured, opaque with 1

Associate Professor (Corresponding author), Department of Veterinary Clinical Medicine, Veterinary College and Research Institute, Namakkal, drvijaymvc@gmail.com. 2Assistant Professor, TVCC, Veterinary College and Research Institute, Thanjavur. 3Assistant Professor and 4Professor and Head, Department of Veterinary Clinical Medicine, Veterinary College and Research Institute, Namakkal <52 < J. Ind. Vet. Assoc., Kerala. 10 (3)

Fig 1. Needle Thoracocentesis

Fig 2. Chyle- White coloured and opaque in nature 3

2.5g/dl protein and WBC 5.23 x10 /cumm). Ultrasonography of the thorax and heart revealed pleural effusion and tricuspid value insufficiency. Electrocardiography and radiography of the animal in standing posture depicted reduced QRS complex and fluid accumulation in the pleura with loss in cardiac silhouette respectively.

TREATMENT The dog was administered with Ringers lactate (10ml/kg body weight intravenous) to counteract shock along with frusemide (4mg/kg IM). Enalapril (0.5mg/kg PO bid), frusemide (4mg/kg PO bid). Salt restricted diet were advocated. it was suggested that the animal should not be put to exercise. Amoxicillin-Cloxacillin (20mg/kg PO bid) was prescribed for five days. The th animal was presented again on 8 day for check up. Dog showed good clinical improvement and was able to take food and water by itself. The degree of dyspnoea was reduced and animal was able to lie down without any respiratory distress. The dog had uneventful recovery from pleural effusion and is presently under treatment with enalapril and frusemide for valvular insufficiency. DISCUSSION Chylothorax is more commonly caused by transmural leakage of chyle that occurred through intact but dilated lymphatic vessels. Underlying diseases that have been reported to cause chylothorax include heart disease (cardiomyopathy, pericardial effusion, heartworm disease, tetralogy of Fallot, tricuspid dysplasia, cortriatriatum dexter), lymphatic or mediastinal neoplasia, fungal granuloma, venacaval thrombosis, peritoneal pericardial diaphragmatic hernia, lung lobe torsion, and congenital abnormalities of thoracic duct (Birchard et. al., 1998). In most small animal patients, chylothorax is considered to be idiopathic because the specific aetiology remains unknown (Ettinger and Feldman, 2010). In the present case, chylothorax is due to heart disease caused by tricuspid valve insufficiency. Sturgess (2001) reported that the common presenting signs in chylothorax were

restrictive breathing pattern, dyspnoea, tachpnoea, coughing, weight loss, muffled heart sound, and decreased lung sound on ventral area. The clinical signs noticed in the present study are similar to the reports of the above author. Chyle appeared white milky to pink coloured, opaque. (Fossum, 2007). Radiography was helpful in identifying masses, neoplasm, cardiac disease or lung lobe torsion. Echocardiography identified cardiac diseases and effusion (Ettinger and Feldman, 2010). Emergency procedure that has to be adopted in patients with pleural effusion is therapeutic needle thoraco-centesis. Needle th thoracocentesis is done at 7 intercostal space on the ventral one third of the thorax and removal of fluid at the rate of 10ml/kg body weight is sufficient to result in significant improvement in respiration (Sturgess, 2001). In the present study, clinical signs, radiography and ultrasonography were useful in diagnosing chylothorax. Other medical management strategies included use of low-fat diets, benzopyrone octreotide in addition to treatment of underlying cause. Benzopyrone may increase the number and function of macrophages to remove protein from lymph and promote fluid reabsorption. Octreotide, a somatostatin analog that inhibits gastric, pancreatic and biliary sectretions, promote gastrointestinal water absorption can also be used (Rasiah et.al.,2003). If chylothorax is idiopathic, surgical management with ligation of thoracic duct with pericardiectomy, pleuro peritoneal shunting, pleuro venous shunting or pleurodesis could be done (Fossum, 2007). SUMMARY A successful management of chyloth-orax due to heart disease in a dog with thoracocentesis, frusemide, and enalapril is placed on record. Vol. 10 Issue 3 December 2012 JIVA <53 <

ACKNOWLEDGEMENT The authors are thankful to the Dean, Veterinary College and Research Institute, Namakkal for the facilities provided.

REFERENCES Birchard, S.J., Smeak,D.D and McLoughlin, M.A. 1998. Treatment of idiopathic chylothorax in dogs and cats. J. Am. Vet. Med. Assoc., 212:652.

Ettinger, S.J and Feldman, E.C. 2010. A textbook of Veterinary Internal Medicine, 7th edition, Saunders Elsevier, Philadelphia. Fossum, T.W. 2007. Small Animal Surgery. 3rd Edition. Mosby Elsevier, St. Louis, Missouri. Rasiah, S.V., Oei, J and Lui ,K. 2003. Octreotide in the treatment of congenital chylothorax. J. Paed. Child Health, 40:585. Sturgess, K. 2001. Diagnosis and Management of chylothorax in dogs and cats. In Practice, 23:506.

HOMAGE TO Dr. VARGHESE KURIEN: INDIA'S WHITE KNIGHT The architect of 'white revolution', Varghese Kurien, who led 'Operation Flood' to transform India from a milk-deficient country to the world's biggest milk producer died on 9-92012 at the age of 90. Hailed as the undisputed 'Milkman of India', who created the billion dollar brand Amul, he is credited with laying the foundation of the nation's co-operative dairy model. The Indian Government had conferred on him the Padma Vibhushan. He was also the recipient of World Food Price, Ramon Magsaysay award for Community Leadership, CarnegieWateler World Peace Prize and International Person of the Year award from US. Born in Kozhikode, Kerala, on November 26, 1921, Kurien had graduated in science from Loyola College in Chennai (1940) and obtained his degree in engineering from the Guindy College of Engineering in Chennai. After a stint at TISCO, Jamshedpur, Kurien got the Government of India's scholarship to study dairy engineering. Following specialised training at Imperial Institute of Animal Husbandry and Dairying in Bangalore, Kurien went to the USA where he completed his masters' degree in mechanical engineering with dairy engineering as a minor subject from Michigan State University in 1948. On his return to India, Dr. Kurien was assigned to join Government Creamery located at Anand in Gujarat to serve his bond period. Kurien then joined Kaira District Cooperative Milk Producers' Union Limited in 1949 on the request of Tribhuvandas Patel, the then Dairy Chairman. The dairy was formed at the initiative of Sardar Vallabhabhai Patel. Later, Patel asked Kurien to help set up a dairy processing plant which saw the birth of Amul. Amul's co-operative model became a success and it was replicated throughout Gujarat. The different dairy unions were later brought under the banner of Gujarat Co-operative Milk Marketing Federation (GCMMF). Dedicating his professional life to empowering the Indian farmers through co-operatives, Kurien, served the GCMMF from 1973 to 2006, and the Institute of Rural Management (IRMA) from 1979 to 2006. Kurien's tenure at Anand changed the destiny of Indian dairy industry. The first dairy co-operative union in Gujarat was formed in 1946 with two village dairy co-operative societies as its members. By 1955, Kurien led to the development of the iconic Amul brand for selling the milk of the co-operative. In 1965, Kurien's leadership caught the attention of the Prime Minister Lal Bhadur Shashtri. He asked Kurien to lead the National Dairy Development Board (NDDB) and replicate the Co-operative success story of Amul across the country. In 1970, with the help of the World Bank, the NDDB started “Operation Flood” which, over the next 26 years, transformed India from a milk importer to world's top most milk producing country. Kurien came to be known as the “Milkman of India” and the “Father of White Revolution". <54 < J. Ind. Vet. Assoc., Kerala. 10 (3)

CLINICAL REPORTS

SURGICAL MANAGEMENT OF TESTICULAR SEMINOMA - A CASE REPORT Laiju. M. Philip1 and M. Ranjith Mohan2 Animal Husbandry Department, Kerala

ABSTRACT Testicular Seminoma was diagnosed in a five year old monorchid German Shepherd dog. After Ultrasoundscanning and laboratory examination, scrotal ablation was done to repair the condition. INTRODUCTION Testicular tumors are common and account for 4 to 7 percent of all tumors in male dogs. Most affected dogs are over 5 years of age, with a median age of 10. The majority of tumors occur in undescended testicles located in the inguinal canal or abdominal cavity. Tumors in descended testicles are less common (Grieco, V. et.al, 2008). The affected testicle is often larger and firmer than its neighbour and has an irregular, nodular surface. At times the testicle is normal size but feels hard. In this paper a case of Testicular Seminoma and its surgical management in a German Shepherd dog is discussed.

CASE PRESENTATION A five year old German Shepherd dog was presented to Veterinary Polyclinic, Mannarkkad 1

Veterinary Surgeon, Veterinary Polyclinic, Mannarkkad

Veterinary Surgeon, Veterinary Dispensary, Karakurissi

with a complaint of scrotal enlargement (Fig. 1). On detailed examination, the animal was found to be monorchid and it was confirmed by Ultrasonography. The scrotal sac was hard in consistency. Ultrasound scanning revealed thickened testicular wall and a homogeneous testicular mass of low echogenicity (Fig. 2). Cytological examination of Fine needle aspirate revealed confirmation of seminoma of testis. Hence decided to perform scrotal ablation. The surgical site was prepared aseptically after restraining the animal. The dog was premedicated with Atropine Sulphate at the rate

Fig. 1 Dog with Enlarged Scrotum 0.45mg/kg body weight intra muscularly, followed by an intravenous bolus injection of Xylazine Hydrochloride at the rate 1 mg/kg body weight and Ketamine Hydrochloride at the rate 5 mg/ kg body Vol. 10 Issue 3 December 2012 JIVA <55 <

Fig. 2 Ultrasound scanning revealing homogeneous testicular mass of low echogenicity weight. An elliptical incision was made around the base of the scrotum. Separated the scrotal skin and fascia to expose the spermatic cord. The testis was removed along with the spermatic cord and scrotal sac, after ligating the testicular artery and testicular vein separately. Apposed the skin incision in a horizontal mattress pattern. Parenteral medication was done using Inj. AmoxicillinDicloxacillin 500 mg for three days. The animal had an uneventful recovery.

There are three common types of testicular tumours. Sertoli cell tumours, seminomas and Leydig (interstitial) cell tumours. Seminoma is a unilateral, single, often benign tumour of the testis; however, malignant forms of the tumor have been reported in rare cases. It is the second most common tumor of the testis in male dogs, typically affecting older dogs (over the age of four). Seminomas develop due to cryptorchidism, a fetal abnormality which occurs when one or both testes fail to descend into the scrotum from where they develop in the abdomen (Hayes, H.M. Jr and Pendergrass, T.W. 1976). There is 16-times risk of developing seminoma in the cryptorchid testicle compared to descended testicle in dogs with unilateral cryptorchidism (Reif, J. S et.al, 1979). Seminoma in descended testicle found in younger dogs and associated with contralateral cryptorchid testicle.Ultrasound examination is a sensitive and relatively specific technique for the diagnosis of testicular tumors.

REFERENCES Grieco, V., Riccardi, E. and Greppi, G.F. 2008. Canine testicular tumours a study on 232 dogs. J. Comp. Path. 138: 86-89. Hayes, H. M. Jr and Pendergrass, T. W. 1976. Canine testicular tumours: Epidemologic Features 410 dogs. Int. J. Cancer. 18 (4) : 482 - 487 Fig. 3. Surgically Removed Testicular Tumour DISCUSSION Testicular tumours affect mostly the Boxer, German shepherd, Afghan hound, Weimaraner and Shetland sheepdog type of breeds. <56 < J. Ind. Vet. Assoc., Kerala. 10 (3)

Reif, J. S., Maguire, T.G., Kenney, R.M and Brodey, R.S. 1979. A. Cohort study of canine testicular neoplasia. J. Am. Vet. Assoc. 175 (7) : 719-723.

CLINICAL REPORT

A REPORT ON THE OCCURANCE OF Hymenolepis anatina IN DUCKS UNDER BACKYARD SYSTEM 1

G. Jyothimol , K. Syamala , M.N. Priya , C.K. Deepa , K.G. Ajithkumar , Ajith Jacob George3 and Reghu Ravindran2 College of Veterinary and Animal Sciences, Pookot

ABSTRACT Two different cases of mortality in ducks reared under backyard system due to severe cestode parasitism were reported. On postmortem examination varying degrees of enteritis was also observed. The parasites were collected and identified based on morphology as Hymenolepis anatina. The present communication reports the occurrence of H. anatina for the first time in domestic ducks of Kerala.

INTRODUCTION Reports on the occurrence of helminth infections in anseriform birds affecting their health and well-being are scant (Schiller,1951). Tapeworms of the genus Hymenolepis have been frequently encountered in large numbers in water fowls reared under intensive conditions (Islam et.al., 1988) . The present communication reports the occurrence of a cestode parasite, Hymenolepis anatina in domestic ducks from Wayanad, Kerala, and its pathogenic effects. 1

MVSc Scholar, 2Assistant Professor, Department of Veterinary

Parasitology, 3Associate professor, Department of Veterinary Pathology, College of Veterinary and Animal Sciences, Pookot, Wayanad

MATERIALS AND METHODS Two cases of mortality in domestic ducks reared under backyard system were reported. Case number 1:- Forty out of 100 ducks of 6 month age reared by a private owner from Mananthavady died. The owner reported that the birds were brought one month before from a poultry market at Thrissur. Case number 2: Three out of 10 ducks owned by an agricultural farmer from Kalpetta showed mortality. In both cases, a representative number of birds were brought to the Department of Veterinary Pathology, College of at Veterinary and Animal Sciences, Pookot, Wayanad for conducting postmortem examination. Parasites were collected for routine processing and identification. Parasites were identified based on (Singh, 2003).

RESULT It was observed that the cestode parasites were present in large numbers as to occlude the lumen of intestine which has resulted in enteritis of varying degree. The parasites were identified as Hymenolepis anatina based on morphological features. Each proglottid showed three testis and one bilobbed ovary within a single segment (Fig.1) Vol. 10 Issue 3 December 2012 JIVA <57 <

Vitelline gland was clearly seen behind the ovary. Genital pore was unilateral in each segment. The scolex revealed the presence of ten hooks on the rostellum, which was typical for H. anatine (Fig.2)

with serious consequences. Among the helminthic infections of anseriform birds, the most commonly observed parasites were of Hymenolepidae family (Schiller,1951). Kharchenko (1960) observed that hymenolepids are most common in summer and spring season. Usually most of the Hymenolepis sp. were considered to be non-pathogenic but severe infections may cause death. In the present study, H. anatina is reported for the first time from domestic ducks of Kerala. There were no available reports on the pathogenicity of these parasites. The mortality observed in ducks could be attributed to the severe cestode parasitism.

REFERENCES Fig.1: Hymenolepis anatina segments Islam, M. R., Shaikh, H and Baki, M. A. 1988. Prevalence and pathology of helminth parasites in ducks of Bangladesh. Veterinary Parasitology. 29:73-77 Kharchenko, O. N. 1960. The problem of helminth - carriers among ducks with hymenolepid infection. J. Helminthologia. 4: 249-253 Schiller, E. L.1951. The cestoda of anseriformes of the North Central states. The American Midland Naturalist. 46: 444-461

Fig.2: Hymenolepis anatina hooks on rostellum

DISCUSSION Cestode parasitism in wild ducks usually maintain a balance, where the host is not affected <58 < J. Ind. Vet. Assoc., Kerala. 10 (3)

Singh, K. R. S. 2003. Veterinary Helminthology. Directorate of information and publication, Indian Council of Agricultural Research, New Delhi. pp: 221

CLINICAL REPORT

MANAGEMENT OF SUBCUTANEOUS EMPHYSEMA IN A HORSE 1

Mir Aamir Ali and H.S. Mahesha

Bangalore Turf Club Limited, Racecourse road, Bangalore.

ABSTRACT A kathiawari horse aged three years was presented with swelling throughout the body. Clinical examination revealed a penetrating wound in the axillary region with subcutaneous emphysema. The wound was closed with antibiotic impregnated gauze with minimal debridement an uneventful recovery occurred.

the body and also involving the face. Clinical examination revealed a penetrated wound in the maxillary region with diffuse accumulation of air with crepitation and the skin pitting on pressure, with congested mucus membranes and anorexia were the other associated symptoms. The wound was cleaned with normal saline and the wound was closed with antibiotic ointment and sterile guaze. Fig. (1and 2).

Keywords: Axillary wound, subcutaneous emphysema INTRODUCTION Axillary wounds are caused by penetration of a sharp object like trees, fences, gates and usually do not involve the thorax (Laverty et.al., 1996). Marked subcutaneous emphysema develops because the wound tissue becomes a one way channel for air and as the horse moves the air progressively accumulates in soft tissues. The trapped air may migrate through fascial planes with potential to cause pneumothorax, (Hance et.al., 1992).

CASE HISTORY AND TREATMENT A kathiawari horse aged about 3 years was presented with a complaint of swelling throughout 1

Assistant Veterinary Officer, 2Veterinary officer, Bangalore Turf Club Limited, No.52, Racecourse road, Bangalore.

Fig.1. Diffusely swollen head and body Vol. 10 Issue 3 December 2012 JIVA <59 <

further accumulation of air and continued until recovery. Surgical closure of the wound was not considered as reported by Marble et.al., (1996), Joellugo (2006) and Florent David et.al., (2008). Antibiotic impregnated gauze was well tolerated by the horse and following which the horse recovered without any respiratory complication.

REFERENCES Fig. 2. Penetrating wound in the axilla

Tetanus toxoid was given initially and gentamicin was given intravenous at 6.6mg per Kg body weight once daily along with metronidazole at 25mg per Kg orally four times daily and penicillin sodium at 22000 I.U. per Kg body weight intravenously twice daily. Phenyl butazone was given at the dose rate of 2.2 mg per Kg intravenously once daily was administered for ten days.

RESULT AND DISCUSSION The clinical symptoms as observed in the current case were also reported by Laverty et.al., (1996), Hassle (2007). Axillary wounds were found associated with subcutaneous emphysema and predisposing to pneumothorax that was not observed in this case. Stall rest was advised to avoid

Florent David and Sheila Laverty, 2008. Thoracic Trauma. In:Five minute Veterinary Consult Equine, Jean pierre Lavoie and Kenneth Hinchcliff (Ed. 2), Elsevier., pp.756-757. Hance, S.R and Robertson, J.T. 1992. Subcutaneous Emphysema from an axillary wound that resulted in pneumomediastinum and bilateral pneumothorax in a horse. J. Am. Vet. Med. Assoc., 200:pp.1107. Hassel, D.M. 2007. Thoracic Trauma in horses. Vet. Clin. Equine., 23:67-80. Joel Lugo. 2006. Thoracic disorders. In Equine Surgery Auer and Stick (Ed. 3), Saunders Elsevier St. Louis USA., pp. 620-621. Laverty, S., Lavoie, J. P., Pascoe, J.R and Ducharme, N. 1996. Penetrating wounds of the thorax in horse. Equine. Vet. J., 28:pp.220. Marble, S.L., Edens, L.M and Shiroma, J.T. 1996. Subcutaneous emphysema in a neonatal foal. J. Am. Vet. Med. Assoc. 208:pp.97.

BEST SCIENTIFIC PAPER AWARD OF INDIAN VETERINARY ASSOCIATION, KERALA Indian Veterinary Association, Kerala has decided to give best scientific paper awards annually for the best papers published in Journal of Indian Veterinary Association, Kerala from the year 2012. The awards will be for a best Clinical paper and a best Research paper. The award consists of a citation and a cash award which will be distributed in the Veterinarians Annual Convention. <60 < J. Ind. Vet. Assoc., Kerala. 10 (3)

GENERAL ARTICLE

IMPACTS OF CLIMATIC CHANGE IN ANIMAL HUSBANDRY- ARE WE PREPARED? S. S. Rani Assistant Director, Livestock Management Training Centre, Kudappanakkunnu, Thiruvananthapuram

ABSTRACT Climatic change and food security are two important emerging global problems. The increasing animal population to meet the food security is also considered as a major catalyst for climatic change and emerging zoonoses. Developing countries are likely to be worse affected by climatic change and difficult to cope with the adaptation or mitigation measures for want of economical and technological support. Species and region specific research are essential to formulate ideal but affordable mitigation measures to meet the climatic stresses. Sensitization of farmers about its importance and capacity building are the immediate needs. Collaboration of multidisciplinary experts and agencies is also important for desired results.

CLIMATIC CHANGE AND GLOBAL WARMING Extreme weather events and its damaging effects are already experienced all over the world. Climatic change is now a reality and the most obvious manifestation is the rising average atmospheric temperature otherwise known as global warming. The mean global annual temperature increased between 0.4 to 0.7째 C.The impacts could be multifarious like rainfall, melting

glaciers, rising sea level, recurrent droughts and floods, threat to biodiversity, increased plant and animal diseases and infinite number of challenges on public health (Sarkar, S. and Padaria, R. N. 2010). The Intergovernmental Panel on Climatic Change (IPCC) predicts the increase in global average surface temperature between 1.8째C and 4.0째 C by 2100 and extinction of approximately 20 to 30 percent of plant and animal species with severe consequences for food security in developing countries (FAO). Effect of Green House Gases (GHG) like Carbon dioxide, Methane and Nitrous Oxide towards global climatic change is already established. At smaller levels these gases have a protective role of life on earth. Carbon dioxide is widely considered the most important humaninduced GHG. Estimated GHG emission from animals is 18 percent, mainly Methane, which got 25 times the Global Warming Potential (GWP) of Carbon dioxide (FAO). Climate-changing gases are released into the atmosphere at every step of meat, egg, and milk production and potentially disrupting weather, temperature, and ecosystem health. Globally emissions from pig manure alone account for almost half of all GHG emissions from farm animal manure. (Koneswaran, G and Nierenberg, D, 2008).

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DIRECT IMPACTS ON ANIMAL HEALTH India is an agrarian country with the largest cattle population in the world. Livestock population have a crucial role in developing countries by its contribution to food security and poverty alleviation. It is also noted that the global livestock sector is growing faster than any other agricultural subsector. But Methane emission from cattle population through enteric fermentation and manure management is a matter for concern in climatic change. Impact of climatic changes could be in four ways: heat-related diseases and stress, extreme weather events, feed grain forage availability and price and adaptation of animal production systems to new environments and emergence or reemergence of infectious diseases, especially vector-borne diseases critically dependent on environmental and climatic conditions. Most important direct effect could be heat stress and reduced nutrient intake affecting production performance (Mariara, J.K., 2008). The results of impact assessment study in Africa indicate that large livestock farms are more vulnerable to climate change and are likely to lose net revenue while small farms are much less vulnerable and will probably get advantage, atleast against the risk of dryness, livestock offer a good substitute for crops. (Seo, S.N. and Mendelsohn. R.2007). It is a requisite to mitigate the climatic stresses, if a high producing and less tolerant animal to be reared in harsh environmental situation. (Naqvi, S.M.K and Sejian, V. 2011). Mitigation to overcome heat stress could be = Physical modification of environment = Genetic development of less sensitive breeds = Improved nutritional management schemes <62 < J. Ind. Vet. Assoc., Kerala. 10 (3)

Some current economic practices such as shades, sprinklers and ventilation will be suitable for adapting to reduce heat stress in dairy cows. Shades are the simplest method to reduce the impact of high solar radiation. Open sided construction for sheds will increase natural ventilation and if not, providing specially designed fans will be ideal. An effective way of cooling cattle is evaporative spray-mist, fog and sprinkling systems. Climate change will have a substantial effect on global water availability in the future. Not only will this affect livestock drinking water sources, but it will also have a bearing on livestock feed production systems. Reduction of livestock numbers to more productive animals leads to more efficient milk/ meat production and lower GHG emissions. Different animal feeding and manure management (collection, storage, spreading, biogas production), management of feed crop production are other mitigation measures. (IFAD, 2007). Emergence/ Re-emergence of diseases: Predictions on all the impacts are difficult but a change in climate can result in changes in species composition of vectors, pathogens and augmenting its spread and even the emergence of new pests and diseases. New transmission modalities and host ranges complicate the epidemiology of some diseases. (FAO, 2008). Temperate countries will be more prone to such diseases. Changes in the spatial distribution of vectors/pathogens, animal populations with little or no immunity would be exposed and suffer major disease impacts (FAO, 2008).Diseases such as Bluetongue have expanded their range due to

global warming. Extreme weather events such as heavy rainfall or droughts often trigger disease outbreaks. PUBLIC HEALTH IMPACTS Climate change is having far-reaching consequences on human health, perhaps most starting signals could be growing conflicts, hunger and then to diseases. (Koneswaran, G and Nierenberg, D, 2008). Climate change may result in food-borne zoonoses. Additional /inappropriate use of pesticides and veterinary drugs could be another concern. Mycotoxins in food is going to be a growing problem in the coming era. Vulnerability of the impact could be more on developing countries where animals and human are exposed directly to natural resources for food and less capacity for adapting mitigation measures. Rural poor communities rely greatly on agriculture and livestock for their survival, at the same time these are amongst the most climatesensitive economic sectors. So climatic change can aggravate poverty. NEED FOR A 'ONE HEALTH' APPROACH It is estimated that up to 75 percent of newly recognised infectious diseases of humans are from zoonotic pathogens- Bovine Spongiform Encephalitis, Nipah virus, Monkey Pox, H5N1, West Nile Virus, Rift Valley Fever and many more to add. It is a clear realisation that infective agents circulate between wildlife, domestic animals and humans. It is also proved that no one discipline or sector of society has enough knowledge and resources to prevent the emergence or resurgence of diseases in today's globalized world. The 'One Health' approach needs contributions from

multidisciplinary experts like agricultural scientists, veterinarians, anthropologists, economists, educators, engineers, entomologists, epidemiologists, microbiologists, nutritionists, physicians, public health professionals, sociologists and local communities (Black, P. F. et.al. 2008) Responses to Climatic changes and future strategies 1. Adaptation to reduce the vulnerability of people and ecosystems 2. Mitigation to reduce the magnitude of impact in the long term. The Conference of the signatories under the United Nations Framework Convention on Climate Change (UNFCCC) have been unsuccessful till date in negotiating consensus on emission reductions. The chances for global agreement in preventing or mitigating climate change are increasingly decreasing. Therefore we need to prepare people for adjusting to the impacts of climate change. Across the globe, governments are increasingly adopting national programmes to deal with the threat of climate change. According to the national action plan on climatic change by the Prime Minister's council, possible risks were analysed with observed impacts and eight National Missions were formulated in strategic areas including a National Mission for sustainable agriculture. Under the aegis of these missions implementation of various activities are in progress. Some of them are conservation of water through practices like rain water harvesting, risk financing like crop/animal insurances, disaster management and proactive programmes on capacity building, surveillance and control of vector borne diseases like Japanese Encephalitis, Vol. 10 Issue 3 December 2012 JIVA <63 <

Dengue fever, Malaria,Filariasis, energy audits in large energy consuming units, promotion of biofuels.( GOI., 2009) Government of Karnataka invited related departments to prepare their action plans, strategies or vision documents for evaluation but only half the agencies could prepare the requisite documents in time. Government of Kerala have the department of Environment and Climatic Change to co-ordinate reduction, adaptation or mitigation measures. METHANE EMISSION REDUCTION STRATEGIES This is very important in India with huge livestock population. Also Methane is the GHG with the shortest life span,10-12 years when compared to CO2 with 120 years. So any effort to reduce methane emission will be beneficial to slow global warming. It could be managemental, nutritional or advanced biotechnological strategies. (Naqvi, S.M.K and Sejian, V. 2011). Methane emission from exotic/crossbred cows with high productivity is proved to be less. Amount of feed consumed and its digestibility are two important factors that determine total Methane production. WHAT NEXT? Identifying task force to formulate policies in relevant sectors, research to develop vulnerability indices on various parameters in the sector and to formulate adaptation strategies that best suited for the farming situation of our state are crucial to meet the hardships of climatic change. Capacity building of staff at various levels to cope

<64 < J. Ind. Vet. Assoc., Kerala. 10 (3)

up with the mitigation measures and sensitisation of farming community about this phenomenon are other immediate interventions required. CONCLUSION Climate change is an ongoing process. Starving in both animals and human can be worse in the developing world. There is a two-way relationship between livestock production and environmental health. On the one hand, livestock contribute to climate change and other environmental problems, and at the same time livestock health and productivity can be adversely affected by these same environmental upsets. (Sherman, D.M.2010). Research should be initiated by responsible forum to formulate policy for sustainable livestock at one end. Developing farmer friendly technologies are important especially in the quality of feed to improve milk production and decrease emission of GHG. Control of emerging diseases need concerted efforts in research, control measures for vector and pathogen including their transboundary movement in the era of enhanced global trade, travel and tourism. Climate determines ecosystem health over time, but weather drives immediate outbreaks and disasters. Hence implications of climatic change on different fields connected with plant, animal and human life need to be studied in depth at different levels for early mitigation. The environmental impacts of animal production require more focussed attention from international organizations, governments, producers, and consumers on meat and dairy production.

REFERENCES Black, P. F., Murray, J. G. and Nunn, M. J. 2008. Managing animal disease risk in Australia: the impact of climate change. Rev. sci. tech. Off. int. Epiz., 27 (2), 563-580 FAO. 2008. Climate related Transboundary pests and Diseases, Technical background document from the Expert Consultation th

held on 25 to 27 February 2008 at FAO, Rome. Available at ftp://ftp.fao.org/docrep/ fao/meeting/013/ai785e.pdf. Accessed on 19-3-2012 GOI. 2009, National Action Plan on Climate Change. Available at india.gov.in/ allimpfrms/alldocs/15651.doc IFAD 2007. Livestock and climatic change. The International Fund for Agricultural Development (IFAD) Strategic Framework 2007-2010. Available at www.ifad.org/sf/ Accessed on 21-3-2012 Koneswaran, G and Nierenberg, D 2008. Global Farm Animal Production and Global Warming: Impacting and Mitigating Climate Change. Environ Health Perspect 116:578582. Mariara, J.K. 2008. The Economic Impact of Global Warming on Livestock Husbandry in Kenya: A Ricardian Analysis. African Economic Conference on Globalization, Institutions and Economic Development of Africa. Tunis, 12-14th November 2008. Available at www.afdb.org .Accessed on 19-3-2012

Naqvi, S.M.K and Sejian,V. 2011. Global climate change: Role of Livestock.Asian J.Agric.Sci,3(1):19-25 Report prepared for Department of Ecology and Environment, Karnataka.Available at www.empri.kar.nic.in/Karnataka. Accessed on 17-3-2012 Sarkar, S and Padaria, R. N. 2010. Farmers' Awareness and Risk Perception about Climate Change in Coastal Ecosystem of West Bengal. Indian Res. J. Ext. Edu. 10 (2) SAT eJournal | ejournal.icrisat.org December. 4 (1). Available at www.icrisat.org/journal/pdf. Accessed on 17-3-2012 Seo, S.N. and Mendelsohn. R. 2007. Climate Change Impacts on Animal Husbandry in Africa: A Ricardian Analysis. World Bank Policy Research Working Paper 4261, June 2007. Available at www.worldbank.org/pdf. Accessed on 20-3-2012. Sherman, D.M. 2010. A Global Veterinary medical Perspective on the concept of one health: Focus on livestock. ILAR Journal. 51(3): 281287. Slenning, B.D. 2010. Global Climate Change and Implications for Disease Emergence. Veterinary Pathology. 47(1) 28-33 State Action Plan on Climate Change Karnataka 2010. Rapid Assessment of Sectoral Actions Thornton, P, Herrero, M, Freeman, A and Mwai, O. 2007. Vulnerability, Climate change and Livestock Research Opportunities and Challenges for Poverty Alleviation

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GENERAL ARTICLE

VACCINES FROM OUR GARDEN Tincy Mary John1, N.M. Shah2, B. S. Chandel3 and H.C.Chauhan4 College of Veterinary Science and Animal Husbandry, SDAU, Gujarat

INTRODUCTION

EDIBLE VACCINES

There is a saying that, “Prevention is better than Cure”. The best way to prevent a disease is to be immunized and the most effective way to get immunized is by vaccination. Thus, Vaccines can said to be a boon in the field of medical science. The world of vaccines can be classified mainly into two groups Classical Vaccines and New Generation Vaccines. Classical vaccines are made up of killed /inactivated or live attenuated microbial agents or bacterial toxoids. With the advancement in the field of biotechnology and molecular biology, we were able to introduce New Generation Vaccines like naked nucleic acid vaccines, vectored vaccines, anti-idiotypic vaccine, recombinant DNA vaccine etc. Still, according to WHO reports, the mortality rate due to illness are increasing every year especially in the third world countries. This may be due to constraints on vaccine production, distribution and delivery. The search by scientists to solve these limitations ultimately results in the development of “EDIBLE VACCINES”.

Edible vaccines are those vaccines based on genetically engineered expression of an antigenic protein by an edible plant. In simpler words, they are simply sub-unit vaccines that are edible in nature. Here, the gene of interest is introduced into plants and then these altered plants are induced to manufacture the corresponding proteins. This process is known as transformation and the altered plants are called transgenic plants. (Sibila, J. et.al.2005)

M.V.Sc Scholar, 2Professor & Head, Department of Veterinary Microbiology. 3Professor & Head, 4Associate professor, Department of Animal Biotechnology, College of Veterinary Science & A.H.,S. D. Agricultural University <66 < J. Ind. Vet. Assoc., Kerala. 10 (3)

The first report of edible vaccine appeared in 1990 in the form of patent application published under the international patent cooperation treaty. (Curtiss, R. I and Cardineau, C.A. 1990) It was regarding the successful expression of Streptococcus mutans surface protein A in tobacco. As this bacterium causes dental caries, it was envisaged that the stimulation of a mucosal immune response would prevent the bacteria from colonizing the teeth and there by protect against tooth decay. (Mason, H. S and Arntzen, C. J. 1995) Later in 1992, Charles Arntzen and his coworkers successfully expressed hepatitis B surface Antigen in tobacco plants in a cost effective manner. This paved the way for worldwide acceptance to the concept of edible

vaccine. (Mason, H. S. et.al.1992) Till then, various attempts have been made to develop edible vaccines against human and animal diseases including Norwalk virus particles, (Mason, H. S et.al.1996) Rabies (Hooper, D.C. et.al., 1994) Gastroenteritis, Cholera (Arakawa, T. 1997).

expressed by this method in experimental animals and plants.

Biolistic method: The gene containing DNA coated metal (e.g. gold, tungsten) particles are fired at the plant cells using gene gun. Those plant cells that take up the DNA are then allowed to grow in new plants, and are cloned to produce large number of genetically identical crop. This method is quite attractive because DNA can be delivered into cells of plant which makes gene transfer independent of regeneration ability of the species. But the chief limitation is that the gene gun is highly expensive.

Electroporation: Here there is introduction of DNA into cells by exposing them for brief period to high voltage electrical pulse which is thought to induce transient pores in the plasma lemma. The cell wall presents an effective barrier to DNA therefore, it has to be weakened by mild enzymatic treatment so as to allow the entry of DNA into cell cytoplasm.

PRODUCTION OF EDIBLE VACCINE For producing Edible Vaccines, any of the two methods can be followed: (Shah, P.C et.al. 2011) 1. Appropriate plant virus is genetically engineered to express the desired peptides/ proteins. The recombinant virus is then inoculated into the plant. Large numbers of new plants are grown and chimeric virions are extracted and purified. The resultant plant edible vaccines are utilized for immunological purpose. 2. In another approach, the gene of interest is integrated with plant vector by transformation. A variety of techniques have been used to introduce transgene into plant cell; these could be grouped into following categories:

Agrobacterium mediated gene transfer: The appropriate gene construct is inserted into the T-region of a disarmed Ti plasmid of Agrobacterium. The recombinant DNA is placed into Agrobacterium; a plant pathogen which is co-cultured with the plant cells or tissues to be transformed. The drawback of this method is that it gives low yield and the process is slow. This method worked especially well for dicotelydenous plants like potato, tomato and tobacco. Studies have also proved that the genes are

â&#x20AC;&#x153;SECOND GENERATIONâ&#x20AC;?EDIBLE VACCINES Scientific community developed the second generation edible vaccine which provides protection against several pathogens. These multicomponent edible vaccines can be obtained by crossing two plant lines harboring different antigens. Yu and Langridge (Jie, Yu and William H.R. Langridge. 2001) fused Cholera toxin (CT) B and A2 subunit complementary DNAs (cDNAs) to a rotavirus enterotoxin and enterotoxigenic Vol. 10 Issue 3 December 2012 JIVA <67 <

Escherichia coli fimbrial antigen genes and transferred into potato. It was found that this trivalent edible vaccine could elicit humoral responses, as well as immune memory B cells and T helper cell responses which are hallmarks of successful immunization.

infection. T-cells produced in the mucous are capable of travelling the mucosal tissues through special 'homing' receptors on their membranes. This means that if an immune response is generated in gastrointestinal lining, T-cells produced there can travel to other mucosal sites, (e.g. the lungs, nasal cavity) providing protection over a large surface area

MECHANISM OF ACTION Mucosal Immune System (MIS) is the first line of defense mechanism of body and thereby the most effective site for vaccination. The most effective route of mucosal immunization is oral route. (Korban, S. S. et.al. 2002). The transgenic plant parts with desired gene are fed directly since the outer tough wall of plant cells acts to protect the antigens against attack by enzymes, gastric and intestinal secretions. This method is known as bioencapsulation. The plant cell wall breaks in the intestines near payer's patches and antigens are released. (Rudzik, R. et.al.1975).These released antigens are taken up by the M-cells and is presented to B cells with the help of antigen presenting cells (APC). The activated B-cells get differentiated into plasma cells and secrete IgA class of antibody and elicit mucosal immunity and humoral immunity. Another important component of mucosal immunity is T-cell mediated immune response where the T-cells specifically recognize pathogens and directly kill the invader themselves. It also helps indirectly to the antibodies to clear <68 < J. Ind. Vet. Assoc., Kerala. 10 (3)

Figure showing mechanism of action in the intestinal mucosa

CLINICAL TRIALS Edible vaccines have been checked for their efficacy in humans. The results of human trials that were tested for different transgenic plants

have showed positive responses and no major safety concerns. ( Streatfield, S.J. 2001). Transgenic plant-made vaccines are also being used in animals. The first edible vaccine to demonstrate efficacy in animal trials was against Transmissible gastroenteritis virus (TGEV) in pigs. Livestock animals are fed with transgenic plants, like Arabidopsis thaliana, alfalfa and potato with antigens to protect them from pathogens; including FMDV, BRV and bovine viral diarrhea virus (BVDV)( Streatfield, S.J. 2001)

we run out, we can simply plant more. Moreover, we can select the local/native crop of a particular area and can engineered it to produce the vaccine. 6. Giving an oral vaccine would require little or no training at all, which reduces the requirement of trained professionals.( Streatfield, S.J. 2001) 7. Eliminates the need of syringes & needles for administration. Needle administered vaccines are plagued with problems of re-use, mis-use and an occasional lack of sterilization. 8. Plants cannot host most human pathogens, so the vaccines will not pose a danger to humans.

ADVANTAGES 1. Trigger mucosal immunity (traditional vaccines may bypass this) which is the body's first line of defense. 2. Adjuvant to enhance immune response is not necessary. 3. Cost-effective in terms of manufacturing, storage and transportation. Edible vaccines are made in molecular farms, and not in multimillion dollar machines. Since most countries have soilrich land, it provides them the convenience in obtaining edible vaccines, and long distance transportation is not required. Maintenance of cold chain not required because plant tissues can be dried or, as when the seeds are targeted, have low moisture content. (Pascual, D. W.2007). Thus the amount spent yearly to preserve vaccine can be saved. 4. Their production is highly efficient and can be easily scaled up. For example, hepatitis-B antigen required to vaccinate whole of China annually, could be grown on a 40-acre plot and all babies in the world each year on just 200 acres of land! 5.

Easily available since produced from plants. If

9. Fear of contamination with animal viruses like the mad cow disease, which is a threat in vaccines manufactured from cultured mammalian cells, can be eliminated. 10. They are subunit preparations containing only the antigen and not any pathogenic genes. Thus it enhances the safety of individual. 11. Multi-component ability of vaccine allows to harbour more than one antigen to prevent many diseases in same individual. (Jie, Yu and William H.R. Langridge. 2001) 12. Antigen can be protected by means of bioencapsulation. 13. They can seroconvert even in presence of maternal antibodies, thus having a potential role in protecting infants against like Group-B Streptococcus, Respiratory Syncytial Virus etc. which are under investigation.

FUTURE CHALLENGES/CONCERNS 1. There is need of proper distinguishing characters to identify between 'vaccine fruit' and 'normal fruit' to avoid misadministration of vaccine Vol. 10 Issue 3 December 2012 JIVA <69 <

which could lead to tolerance. 2. Antigen selection involves safety concerns of whether or not selected antigens are compatible enough with the selected plant type to be expressed (Lal, P. et.al. 2007) 3. How can the vaccine dose be controlled? This remains the most difficult task. Consistency of dosage form differs from fruit to fruit, plant to plant and generation to generation. It is determined by protein content, patient's age, weight, ripeness of the fruit and quantity of the food eaten. If low doses are consumed then the production of antibodies are less likely to occur, and if high doses are consumed it may cause tolerance. (Lal, P. et.al. 2007) 4. Glycosylation patterns in plants differ from those in humans and could affect the functionality of the vaccines. 5. Allergic reactions to the fruit or vegetable expressing the foreign antigen may be an issue. 6. The doubt still exists about whether the antigens be able to survive the hostile, acidic conditions of the stomach and even if they did, will they be able to trigger the immune system in the right way. Although initial trials have shown promising results in human subjects, it is not clear what will happen when the person comes in contact with the actual virus. 7. Certain foods are not eaten raw (e.g. potato) and needs cooking which will denature or weaken the protein present in it. (Moss, W. J. et.al.1999) 8. Variable storage conditions for edible vaccine are also a major problem. Potatoes containing vaccine could be stored for longer time while a tomato does not last long. Thus these <70 < J. Ind. Vet. Assoc., Kerala. 10 (3)

vaccines need to be properly stored to avoid infection through microbial spoilage. 9. Another concern is if the transgenic plants are mass produced, they may have an inconsistent expression caused by the small interfering RNAS. (Tonks, A. 2007) 10. There is chance of plant/crops (food) contamination through cross pollination and of vaccine itself in plant debris spreading dust and other pollutant in surfaces and ground waters. The vaccine antigen may affect browsing animals and humans living in the area drinking vaccine polluted water or breathing vaccine polluted dust. The cultivation and production should be limited to facilities like greenhouse, or in plant tissue culture that prevent their environmental release.

CONCLUSION Transforming plants to carry vaccines is one of the latest innovations of medical technology and promises greatest hope for the floundering biotech industry. It can prove to be very effective, if rightly implemented, in providing accessibility to developing and underdeveloped countries where rates of diseases are relatively high. Edible vaccines are prominent over typical traditional vaccines due to its positive aspects like they are cost-effective, safe, easy to administer and can store at the site of production. But it has to overcome the above mentioned concerns and technical obstacles to become a reality. Future research is also required to demonstrate whether these vaccines meet the standards of quality (purity, potency, safety, efficacy and durability) defined for vaccines by the World Health Organization. It can give us new and dramatic hope for improved life. Although in the initial stages of development, a day is not away

when we will be able to pluck a fruit from the garden, munch on it and get protected from diseases… making needles needless… As Hippocrates said, “Let thy food be thy medicine”

REFFERENCES Arakawa, T., Chong, D. K and Merritt, J. L. 1997. Expression of cholera toxin B subunit oligomers in transgenic potato plants. Transgenic Res. 6:403-13. Hooper, D .C ., Pierarrd, L., Modelska , A., Otvos, L. J and Fu, Z. F .1994. Rabies ribonucleocapsid as an oral immunogen and immunolo-gical enhancer. Proc. Natl. Acad. Sci. USA; 91:10908-912. Jie, Yu and William, H.R. Langridge. 2001. A plantbased multicomponent vaccine protects mice from enteric diseases. Nature Biotechnology.19:548-552 Korban, S. S., Krasnyanski, S. F and Buetow, D. E. 2002. Food as production and delivery vehicle for human vaccine, J. Am. Coll. Nutr.21,3 Suppl 2125-2175

Mason, H. S., Ball, J. M and Shi, J. J. 1996. Expression of Norwalk virus capsid protein in transgenic tobacco and potato and its oral immunogenicity in mice. Proc. Natl. Acad. Sci. USA; 93:5335-40. Moss, W. J., Cutts, F and Griffin, D .E. 1999 Implications of human immunodeficiency eradications of measles. Infect. Dis. 29:4452-4455 Pascual, D. W. 2007. Vaccines are for dinner. Proc. Natl. Acad. Sci. USA 104: 10757-10758. Rice, J., Ainley, W.M and Shewen, P. 2005: Plantmade v a c c i n e s : b i o t e c h n o l o g y a nd immunology in animal health. Anim. Health Res. Rev. 6:199-209 Rudzik, R., Claney, R. L, and Perey, Y .E. 1975. Repopulation with IgA containing cells of bronchial and intestinal lamina propria after of homologous payer's patch and bronchial lymphocyte. J. Immunol.144:1599 Shah, P.C., Trivedi, N.M. , Vachhani, D.U, and Joshi, J.V. 2011. Edible Vaccine: A Better Way for Immunization. Int. J. Curr. Pharm. Res. Vol. 3, Issue 1. 53-56.

Lal, P., Ramachandran, V. G., Goyal, R and Sharma, R. 2007: Edible vaccines: Current status and future. Indian .J. Med. Microbiol. 25:93102.

Sibila, J., Snjezana, M and Natasa, B. 2005. Production of biopharmaceuticals, antibodies and edible vaccines in transgenic plants. Current studies of biotechnology. Vol. IV.121-127

Mason, H. S., Lam, D. M. K and Arntzen C J. 1992. Expression of hepatitis B surface antigen in transgenic plants. Proc .Natl. Acad .Sci. USA. 89:11745-49.

Streatfield, S.J. 2001: Mucosal immunization using recombinant plant-based oral vaccines. Methods, 38:150-157.

Mason, H. S and Arntzen, C. J. 1995.Transgenic plant as vaccine production system. Trends Biotechnol. 13:388-92.

Tonks, A. 2007: Oral vaccines: Spoonful of antigens. Br. Med. J, 335:180-182.

Vol. 10 Issue 3 December 2012 JIVA <71 <

GENERAL ARTICLE

“BIODIESEL PRODUCTION FROM ANIMAL FATS” AN EVER GREEN TECHNOLOGY FOR THE FUTURE ENERGY SECURITY John Abraham1 and Ramesh Saravana Kumar2 College of Veterinary and Animal Sciences, Pookode, Wayand.

INTRODUCTION The world is on a quest for energy, the premier source of which is now petroleum. Energy is the most fundamental requirement of every nation as it progress through the ladder of development. India with 16 percent of the world population could boast of only 0.5 percent of the world oil reserves. The current level of production barely caters to 26 percent of the petroleum demand. India imports 75 percent of its crude oil requirements. The estimated crude oil import cost comes to about 10 percent of the country's G.D.P. Besides the billions spent on importing crude oil, we are also importing billions of tonnes of CO2 and other green house gases causing climatic changes as evidenced by changing rainfall patterns, rising sea levels and temperatures. The demand for diesel is five times higher than the demand for petrol in India and any increase in diesel price immediately pushes up inflation. Therefore, the government is still incurring huge cost of subsidising diesel. With the Indian economy poised for a robust growth of 9 to 9.5 percent for the 12th plan period, energy security has become the key issue in policy formulation and planning. 1

Assistant Professor, Department of L.P.M, College of Veterinary and Animal Sciences, Pookode, Wayanad. 2

Professor and Head, Department of L.P.M, Veterinary College and Research Institute, Namakkal, Tamil Nadu. <72 < J. Ind. Vet. Assoc., Kerala. 10 (3)

India's energy security would remain vulnerable until alternative fuels to substitute/ supplement petro-based fuels are developed based on indigenously produced renewable feedstocks. In biofuels, the country has a ray of hope. Biofuels are non-polluting and virtually in exhaustible. Biofuels can increasingly satisfy these energy needs in an environmentally benign and cost-effective manner while reducing dependence on import of fossil fuels and thereby providing a higher degree of National Energy Security (Anon. 2009). The national biofuel policy 2009, proposed 20 percent blending of biodiesel by 2017. This huge demand cannot be met from nonedible vegetable oil feedstock alone. Therefore, development and utilization of new indigenous biomass feedstock for production of bio fuel and development of next generation of more efficient bio fuel conversion technologies are the need of the hour. In this context, biodiesel production from animal fats offers new scope as a potential means to stimulate rural development, lower emission of harmful pollutants and decrease green house gas emission, while contributing to national energy security by reducing dependence on oil imports and mitigation of climatic changes vis-à-vis providing good fuel properties for the diesel engine.

Definition of Biodiesel Biodiesel is defined as â&#x20AC;&#x153;the mono alkyl ester of long chain fatty acids derived from renewable lipid feed stock such as vegetable oil or animal fats, for use in compression ignition (diesel) enginesâ&#x20AC;? (National Biodiesel Board, 1996)

the almost zero sulphur content and the reduced amount of carbon monoxide, unburned hydrocarbons and particulate matter in the exhaust. BIODIESEL PRODUCTION TECHNOLOGY Biodiesel Feedstock

Blends Blends of biodiesel and conventional hydrocarbon based diesel are products most commonly distributed for use. Much of the world uses a system known as the "B" factor to state the amount of biodiesel in any fuel mix: =

100 percent biodiesel is referred to as B100, while

20 percent biodiesel is labelled B20

5 percent biodiesel is labelled B5

Blends of 20 per cent biodiesel with 80 per cent petroleum diesel (B20) can generally be used in all diesel engines without any modification. Biodiesel in its pure form (B100), may require certain engine modifications to avoid maintenance and performance problem. Importance of Biodiesel Biodiesel has good fuel properties, comparable to or even better than petroleum diesel. It has 10 percent built-in oxygen content that helps it to burn fully. Its cetane number (an indication of its fuel burning efficiency) is 72 for biodiesel derived from tallow and 72.5 for biodiesel derived from chicken oil, higher than 54.4, the cetane number of most petroleum diesels. The esters of the long-chain fatty acids of biodiesel are excellent lubricants for the fuel injection system. It has a higher flash point than diesel, making it a safer fuel. Other advantages are

Biodiesel is typically made from vegetable oil though animal fat can also be used. Rapeseed oil has 82 percent of the share of the world's biodiesel feedstock, followed by sunflower oil (10 percent), soy bean (5 percent) and palm oil (3 percent). The choice of feed is country specific and depends on availability. Other feedstocks include waste vegetable oil (WVO), algae which can be grown using waste material and oil from halophytes such as Slicornia bigelovii. Biodiesel Production Process The major steps required to synthesise biodiesel are as follows 1. Feed stock pre-treatment : If waste vegetable oil (WVO) is used, it is filtered to remove dirt, charred food, and other non-oil material often found. 2. Determination and treatment of free fatty acids (FFA) : A sample of the cleaned feedstock oil is titrated with a standardised base solution in order to determine the concentration of free fatty acids (carboxylic acid) present in the oil sample. 3. Transesterification: Transesterification (also called alcoholysis) is the reaction of fat or oil with an alcohol to form esters and glycerol. The reaction is as shown in the equation below. Vol. 10 Issue 3 December 2012 JIVA <73 <

Alkali Catalysed Process

Where R1, R2 and R3 are long hydrocarbon chains sometimes called fatty chain.

Alkali-catalyzed transesterification is much faster than acid-catalyzed transesterification and is most often used commercially. For an alkalicatalyzed transesterification, the glycerides and alcohol must be substantially anhydrous (Wright et.al., 1944) because water makes the reaction partially change to saponication, which produces soap. Low free fatty acid content in triglycerides is required for alkali-catalyzed transesterification.

Alcohol Alcohols are primary and secondary monohydric aliphatic alcohols having 1-8 carbon atoms (Sprules and Price 1950). Among the alcohols that can be used are methanol, ethanol, propanol, butanol and amyl alcohol. Methanol is most frequently used because of its low cost and its physical and chemical advantages (polar and shortest chain alcohol).

Alkali/ Base catalyst The alkalis include Sodium hydroxide, Po t a s s i u m h y d r o x i d e , C a r b o n a t e s a n d corresponding Sodium and Potassium alkoxide such as Sodium methoxide, Sodium ethoxide, Sodium propoxide and Sodium butoxide.

Acid Catalyst Sulphuric acid, Sulfonic acids and Hydrochloric acid are used as acid catalyst.

Lipase Enzyme Lipases can also be used as bio-catalysts.

<74 < J. Ind. Vet. Assoc., Kerala. 10 (3)

Acid Catalysed Process Acid catalysed process are used for direct esterification of free fatty acids in a high free fatty acid (FFA) feed stock, or to make esters from soap stock (Keim, 1945).

Multiple Step Process Canakci and Van Gerpen (2001) investigated an acid catalyzed pre-treatment step followed by a base catalyzed step as an effective conversion method for low cost raw materials. They found that by using methanol and sulfuric acid and a reaction temperature of 60째C the free fatty acid content of a feedstock could be significantly reduced. The preferred method for high FFA feed stock is acid catalysis followed by base catalysis.

Ultrasonic-Reactor Method In the ultrasonic reactor method, the ultrasonic waves cause the reaction mixture to produce and collapse bubbles constantly. This cavitation provides simultaneously the mixing and heating required to carry out the transesterification

process. Thus using an ultrasonic reactor can drastically reduces the reaction time, reaction temperatures, and energy input.

Micro-Wave Method Current research is being directed into using commercial microwave ovens to provide the heat needed in the transesterification process. The microwaves provide intense localized heating that may be higher than the recorded temperature of the reaction vessel.

ALTERNATE PROCESSING TECHNIQUES Enzymes Enzymes have shown good tolerence for the free fatty acid levels of the feedstock, but the enzymes are expensive and unable to proceed the reaction to completion. Immobilisation of enzymes and use of multiple enzymes in sequence may provide future opportunities in this area. (Nelson et.al., 1996)

Co-Solvent Boocok et.al., (1998) developed a novel technique for accelerating the transesterification reaction rate. He proposed the addition of cosolvent to create a single phase and this accelerates the reaction. The commonly proposed co-solvents are Tetra hydro furan and Methyl tertiary butyl ether. The major draw back of this method are the hazard level associated with the co-solvent and recovering and recycling the co-solvent.

Supercritical Methanol System By increasing both the temperature and

the pressure, a critical point is obtained at which gasses and liquids are indistinguishable fluids. Matter that exists in this new phase is called a super critical fluid (SCF). Methanol at supercritical conditions becomes an excellent solvent and dissolves the feedstock and reacts readily without a distinct catalyst, but energy costs of production are similar or less than catalytic production routes. (Saka and Kudsina, 2001).

Animal Fats for Biodiesel Production The amount of animal fat used by the biodiesel industry has more than doubled from 2006 to 2008. According to a report by the Informa Economics, approximately 20 percent of the biodiesel manufactured in the United States in 2008 was produced from animal fats greases and recycled cooking oils. Whole dead poultry carcasses and poultry slaughter wastes can be used to recover fats and oils. The dead poultry birds having an average body weight of 1.25 kg have a fat content of 14.55 percent and by solvent extraction method 96.1 percent of this fat can be recovered. A two step processing reaction, acid catalysed esterification of FFA followed by base catalysed transesterification of triglycerides could convert 97.62 percent of this fat to good quality biodiesel. A litre of biodiesel can be produced from chicken fat at the cost of 22.00 rupees. The blending of biodiesel to commercial diesel at 20 percent can substantially reduce the smoke (47.14 percent) compared to that of diesel in a CRDI engine (John Abraham, 2012). Thus the blending of biodiesel at 20 percent to commercial diesel can reduce the import of costly crude oil and simultaneously, substantially reduce the engine emission. The total weight of the dead birds Vol. 10 Issue 3 December 2012 JIVA <75 <

available in India per year was estimated at 2.4 lakh tons as per the standard mortality of the industrial average (Chandrasekaran, 2009). Which can be converted to 8500 tonnes of biodiesel. Thus biodiesel production technology provides opportunity to produce highly valued biofuel from dead animal and birds. (Wealth from waste). At the same time, this concept can also solve the major problem of unscientific disposal of dead birds and slaughter waste. Currently a 5 million dollar plant is being built in the USA, with the intent of producing 11.4 million litres of biodiesel from an estimated one billion Kg of chicken fat produced annually at the Tyson poultry plant. Rearing animals for meat and biodiesel can be the production objectives in days to come. This concept can revolutionise animal production sector. Especially pigs and poultry can contribute in a big way because of their excellent attributes such as prolificacy, short generation interval and quick body weight gain for slaughter. Fat less pork at a premium price is gaining consumer acceptance phenomenally. This provides the separated fat to be converted to biodiesel economically.

Conclusion The biofuel industry is poised to make important contribution to meeting India's energy needs by supplying clean domestic fuel. Simultaneously it also provides other advantages like 4 It provides a market for excess production of vegetable oils and animal fats. 4 It decreases, although will not eliminate the country's dependence on imported crude oil. <76 < J. Ind. Vet. Assoc., Kerala. 10 (3)

4 Biodiesel in renewable and does not contribute to global warming due to its closed carbon cycle. 4 By biodiesel blending the overall carbon dioxide emission can be reduced by 78 percent compared with petroleum based diesel fuel. 4 The exhaust emissions of carbon monoxide, unburned hydrocarbons and particulate emissions from bio-diesel are lower than with regular diesel fuel. Unfortunately there might be a slight increase in oxides of nitrogen (NOx) 4 When added to regular diesel fuel it can convert fuel with poor lubricating properties, such as modern ultra-low sulphur diesel fuel into an acceptable fuel. 4 Provide good fuel properties for the diesel engines. Biodiesel is not a replacement technology; it is a transition technology to help clean up our existing fuel and stream line distribution by keeping the manufacture and consumer of fuel as local as possible, all the while keeping the revenue in the national economy. It is not about replacing all of our petroleum imports; it is about not importing so much.

REFERENCES Anon. 2009. National Policy on Bio-fuel. Government of India, Ministry of New and Renewable Energy. Boocock, S.K., S.K. Konar, V. Mao, C. Lee and S. Buligan. 1998. Fast formation of high purity methyl esters from vegetable oils, JAOCS, 75: 1167-1172.

Canakci, M and Van Gerpen, J. 2001. Biodiesel Production from Oils and Fats with High Free Fatty Acids. Transaction of the Am. Soc. Agric. Eng., 44 (6): 1429-1436.

Keim, G. I.1945. Testing fat and fatty oils, U. S. Patent No. 2,383,601. Nelson, L.A., Foglia, T.A and Marmer, W.N. 1996. Lipase catalyszed production of bio-diesel. JAOCS, Vol. 73, no.8.

Chandrasekaran, D. 2009. Poultry waste management- Situation in India. Paper presented at the national seminar on wealth from livestock waste, 12-13 November 2009. Veterinary College and Research Institute, Namakkal, Tamil Nadu.

Saka, S. K. Kusdiana. 2001. Biodiesel fuel from rapeseed oil as prepared in supercritical methanol, Fuel 80, (2001), 225-231.

John Abraham. 2012. Utilization of dead poultry birds for biodiesel production. Ph.D thesis submitted to the Tamil Nadu Veterinary and Animal Science University, Chennai.

Wright, H. J., Clark, J.B and Coburn, H.V. 1944. A report on ester interchange. Oil and Soap 21,145-148.

Sprules, F. J and Price, D. 1950. Production of fatty esters. U. S patent 2, 366-494.

Vol. 10 Issue 3 December 2012 JIVA <77 <

ASSOCIATION NEWS

KERALA VETERINARY SCIENCE CONGRESS 2012 released the compendium of the science congress which was received by Dr. K G Suma, Director of Animal Husbandry. Dr. Arunkumar, President, IVA Kerala presided over the function. Dr. K Udayavarman, President, KSVC & Director of Museum & Zoo, Dr. Ani S Das, MD, KFL Ltd. addressed the gathering. Dr. K C Prasath, President KVSSA, and Dr. E K Easwaran, President AHOAK offered felicitations. The participants were welcomed by Dr. Theodore John, General Convenor. The Kerala Veterinary Science Congress 2012 was organised by Indian Veterinary Association, Kerala in association with Dept. of Annual Husbandry, Kerala State Veterinary Council, KLD Board, KSPDC and HLL Life Care Ltd on 10th and 11th Nov 2012 at Trivandrum. The Science congress provided and effective platform for veterinary practitioners, researchers and academicians to present their works and to interact among themselves as well as with the peers of the noble profession. Over 200 delegates from across the country participated in the science Congress. The Science congress was inaugurated by Dr Renukaprasad, Honâ&#x20AC;&#x2122;ble Vice Chancellor of Karnataka Veterinary, Animal & Fisheries Sciences University. Dr. Radhakrishna Pillai, Director, Rajiv Gandhi Centre for Biotechnology, Trivandrum

In the Scientific session that followed papers were presented in 3 session viz. Animal Health Science, Animal Production and management and Basic Veterinary subjects and field veterinary experience. Dr. Harikrishna Kumar, SVS, VH, Aryanad, Dr. Aravind, SVS, DVC, Kollam and Dr. Geetharani, VS, Kollam were winners of best paper/ poster presented in session 3 comprising of field veterinary experience. The lead papers in each session was given by Dr. Kumanan, Director of Reserach TANUVAS, Dr. Sreenivasan, Director of Distance Education, TANUVAS and Dr. Ani S Das, MD, KFL Ltd. The valedictory Address was given by Dr. Sreenivasan, Director of Distance Education, TANUAS and Dr. N N Sasi, Registrar of Kerala State Veterinary Council gave away the prizes to the winners.

KVSSA LEADERSHIP TRAINING CAMP 2012 KVSSA leadership training camp was held on 6th and th 7 October 2012 at N C Gardens Beach Resort, Parappanangadi, Malappuram, Dr. K. R. Arun Kumar, IVA President inaugurated the training camp at a function presided by Dr. K. C. Prasath, KVSSA President. Dr. Joby George, General Secretary, KVSSA welcomed the participants and Dr. K. A. Sajeeev Kumar, Dr. Ajaykumar felicitated. Dr. Sivadas K. T proposed the vote of thanks. The faculty members for the camp were Mr. M. N. Chandran Nair and Mr. Subramanian (International trainers of JCI). 30 Veterinarians participated in the camp. J. Ind. Vet. Assoc., Kerala. 10 (3) <78 <

WORLD RABIES DAY COMMEMORATION

Indian Veterinary Association, Kerala organized a technical seminar on rabies eradication at Hotel National Park, Ettumanoor on 28-09-2012. The function was inaugurated by the Honâ&#x20AC;&#x2122;ble Finance Minister, Sri. K M Mani, Dr. Arunkumar, IVA, Kerala State President presided over the function. Dr. C Sreekumar, General Secretary, IVA Kerala welcomed the gathering. The delegates were veterinary surgeons deputed from all the 14 district of Kerala. Dr. Ilona Otter, DVM, Director, World Wide Veterinary Services India (WVS) handled the highly informative technical session. Sri. K M Mani in his inaugural address stressed the need for humane treatment and handling of animals. He noted that Animal Birth Control (ABC) of dogs and Rabies vaccination are the correct and most humane way to control human rabies cases. He said killing of dogs is not an accepted method to control rabies anywhere in the world. The fear of rabies is what makes society so critical of stray dogs. He lauded the efforts of IVA, Kerala in addressing this critical issue by conducting awareness campaigns, implementation of ABC programme in institutions cases throughout the district of Kottayam and sponsoring vaccines free of cost for pet dogs and cats in the run up to the commemoration programme. The Finance Minister

accepted a memorandum and acknowledged the high risk veterinary doctors face and announced that risk allowance will be sanctioned to all cadres of doctors working in AHD.

ABC SURGICAL CAMP

As part of the World Rabies Day commemoration, IVA Kottayam unit has conducted nearly 60 animal birth control surgeries (spaying) for free in all taluks of the kottaym district. The association also conducted free anti-rabies vaccination programmes in all veterinary hospitals with the cooperation of its members. IVA state unit congratulates the office bearers and the members of Kottayam unit for taking up such an ethical initiative which is first of its kind in the country. Vol. 10 Issue 3 December 2012 JIVA <79 <

SIGN POST & ACHIEVEMENTS Dr. P. Rajeev, District Epidemiologist, ADCP, Alappuzha (1989 batch, COVAS, Mannuthy), received ''1st DAN BLACK BELT “ in “ TAEKWONDO” from “ KUKKIWON” ( World Taekwondo Headquarters ), South Korea. Taekwondo is a Korean martial art and a recognized Olympic game. He got training at the Masters Taekwondo Federation, Pathirappally, Alappuzha, Kerala and spent many years for this marvellous achievement. The black belt test was conducted by Asia’s one and only Olympic referee Mr Maradi Chandrasekhar who holds 7th DAN Blackbelt in Taekwondo Dr. Jacob Alexander, Veterinarian of Thiruvananthapuram zoo and his team was honoured for the timely diagnosis and successful treatment of Trypanosomiasis by awarding “Ponnada” on 26/07/ 2012 at a function held at the Zoo. Ministers K.B. Ganesh Kumar, P. K. Jayalakshmi, K. Muraleedharan, MLA, and District Governor of Lions Club International P. Surendran were among the dignitaries present on the occasion.

OBITUARY Dr. V. Sathianesan (75), Former Professor and Head, Department of Veterinary Parasitology, CVAS, Mannuthy left for his heavenly abode on 02.12.12. Dr. Sathianesan was an authority in Helminthology, especially gastro intestinal nematodes in ruminants. He was always keen in the activities of the Indian Association for the Advancement of Veterinary Parasitology (IAAVP) and had been the member of its Executive committee. He is survived by his wife, Smt. Saralamani and two daughters, Anila and Anupa.May his soul rest in peace. Dr H. Sivasubrahmaniam Addl. Director (Rtd) passed away on 21/10/2012 at Trivandrum. He was regarded as scholar of the profession and was a well wisher of the Veterinary Community. He contributed his personal collection of books to IVA Library. May his soul rest in peace.

Dr. C.N. Ramachandran Nair (71), Addl. Director (Rtd) passed away on 25/09/ 2012 . He served the Animal Husbandry Department in various offices and got retired from service as Addl. Director. May his soul rest in peace Printed at : Micro Printers, Mannarkkad, Phone: 04924-224318 J. Ind. Vet. Assoc., Kerala. 10 (3) <80 <