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ThesisItem Open Access MARKER ASSISTED SELECTION FOR MILK PRODUCTION TRAITS IN VECHUR CATTLE(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2005) SHYMAJA UTHAMAN; K.V. RaghunandananA study was conducted in Vechur cattle of Kerala to study the association of microsatellite marker alleles with the milk production and composition traits and to evaluate seven ancestors using microsatellite markers. Blood and semen was used as the source of DNA in this study. DNA samples were extracted from fifty unrelated animals and thirty three ancestors and progenies. Five microsatellite markers with established linkage to QTL controlling milk production traits like milk yield, fat percentage and protein percentage were used. The DNA samples obtained from the fifty unrelated Vechur cattle were typed using these five microsatellite markers. The five markers were found to be highly polymorphic with the PIC value ranging from 0.82 at the BM1508 locus to 0.89 at the HUJII77 locus. Data regarding the peak yield, days to attain peak yield, AFC and intercalving period were collected from the Vechur Conservation Centre, Mannuthy. Milk samples collected from the Vechur Conservation Centre were used to determine the milk fat percentage, protein percentage, SNF and total solids. The mean fat percentage and AFC were found to differ significantly in different families. Statistical analysis revealed that the V43 and V88 families had a mean fat percentage significantly greater than the population mean. The mean AFC of V43 family was also found to be significantly greater than the population mean. The V44, V88 and VI17 families had a mean AFC significantly lower than that of population mean. The V44 family had the lowest mean AFC and was also significantly different from the V88 and VI17 families. Microsatellite analysis was carried out for the thirty three combinations of ancestors and progeny using the same five markers. The allelic effect of microsatellite markers in the families was studied for fat percentage and AFC. The allelic mean for fat percentage of 223 bp size allele at the HUJII77 locus was found to be significantly higher when compared to those animals without this allele even though there were no earlier reports indicating linkage between this marker and QTL affecting fat percentage. The allelic mean of fat percentage for 205, 209 and 221 bp at the HUJII77 locus was significantly lower when compared to those of animals without this allele. The mean of the animals with 208 bp allele at the ILSTS096 locus was found to be significantly greater for fat percentage when compared to the mean of those without this allele. There are earlier published reports indicating linkage between this marker and QTL affecting fat percentage. Selection for this allele will not be effective as it is already present in high frequency in the population. The 252 bp allele at the BL41 locus had an allelic mean for fat percentage significantly lower than that for animals without this allele. The 238 bp size allele of this locus had an allelic mean for AFC significantly greater than that for animals without this allele. Earlier reports establish linkage between this marker and QTL affecting fat percentage but not AFC. Selection can be applied against this allele to prevent its frequency from increasing in the population. The allelic mean of 158 bp allele at the BM1508 locus had a mean fat percentage significantly lower than that for the animals without this allele. There are earlier reports of linkage of this marker with QTL affecting protein percentage but not fat percentage. Selection against this allele is necessary as this allele is present in high frequency in the population. Animals with 113 bp allele had an allelic mean of fat percentage significantly lower than that for the animals without this allele. This marker has linkage with QTL affecting milk fat percentage. It is advisable to select against this allele to increase the milk fat percentage in the population. Even though the V44 sire can be selected for improving the fat percentage, the mean AFC of this sire was found to significantly greater than that of the population mean hence it is not preferred. The sire V88 can be selected for improving the fat percentage as well as reducing the AFC. There is a possibility to improve the Vechur cattle maintained at the Vechur Conservation Centre, Marmuthy for fat percentage and AFC by adopting line breeding for V88 sireThesisItem Open Access GENETIC POLYMORPHISM OF MILK PROTEINS IN GOATS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 1996) T. V. RAJA; B. NandakumaranThe present investigation was undertaken to identify the biochemical polymorphism at different milk protein loci and to establish their inheritance pattern in Malabari and its exotic cross viz., Alpine x Malabari. It was also envisaged to analyse the association of different milk protein variants with lactation milk yield. Hundred goats belonging to Malabari (50) and Alpine x Malabari (50) were typed for milk protein variants employing horizontal Polyacrylamide Gel Electrophoresis (PAGE). The milk protein loci studied were Alpha Si casein, Alpha Sj casein. Beta casein. Kappa casein and Beta lactoglobulin. Two Alpha Si casein variants, the faster Alpha Si casein F and the slower Alpha Si casein S with three phenotypes Alpha Si casein AA, Alpha Si casein AB, and Alpha Si casein BB were observed. S allele had the frequency of 0.52 in Malabari and 0.70 in Alpine x Malabari crossbreds. The gene frequency of Alpha Si casein F and Alpha Si casein S were 0.48 and 0.52 in Malabari and 0.30 and 0.70 in Alpine x Malabari crossbreds, respectively. The frequency of S allele was higher in both the populations. Alpha Sj casein locus exhibited three phenotypes viz., AA, AB and BB controlled by two alleles A and B. A allele had the ii higher frequency of 0.71 in Alpine x Malabari crossbreds compared to 0.62 in Malabari. Higher frequency of AA phenotype was found in Alpine x Malabari (0.52) and that of AB phenotype in Malabari (0.44) . Two alleles namely A and B contributed three phenotypes viz., AA, AB and BB at the beta casein locus in both the populations studied. Malabari showed higher frequency of B allele (0.55) while Alpine x Malabari had higher frequency of A allele (0.60) . Beta casein AB phenotype had higher frequency in Malabari breed (0.54) . Kappa casein locus showed two alleles A and B contributing three phenotypes viz. , AA, AB and BB. The frequency of A allele was higher in both the population. The phenotype AA was higher in Alpine x Malabari crossbreds (0.56) while Malabari had higher frequency of AB phenotype (0.48) Two alleles namely A and B with three phenotypes AA, AB and BB were identified at Beta lactoglobulin locus. The phenotype AA was dominant in Alpine x Malabari crossbreds (0.50) and the frequency of BB phenotype was almost similar in both Malabari and Alpine x Malabari crossbreds (0.14 in Malabari and 0.12 in Alpine x Malabari crossbreds). The gene frequency of Beta Ig A was found to be 0.65 in Malabari and 0.67 in Alpine x Malabari crossbreds. iii Both the populations studied were in genetic equilibrium with respect to these five milk protein loci. No significant diversity was found to exist between genetic groups. The genetic variability in the populations was calculated by estimating the heterozygosity. The overall heterozygosity in different populations indicated that the Malabari breed had comparatively higher degree of heterozygosity (0.4711) followed by Alpine x Malabari crossbreds (0.4209) . In Malabari maximum heterozygosity was observed at Alpha Si casein loci (0.4992). In Alpine x Malabari maximum heterozygosity was observed at Beta casein locus (0.4800) . The study of association between the genes controlling synthesis of milk proteins showed significant association between the Beta casein with the Beta lactoglobulin BB types in Alpine X Malabari crossbreds. All the other combinations of genes did not show any significant association. Milk protein variants were not found to be associated with the lactational yield (120 days) of the goats. The present study could establish the existence of biochemical polymorphism at Alpha Si casein, Alpha Sj casein, Beta casein. Kappa casein and Beta lactoglobulin loci in Malabari and Alpine x Malabari crossbreds studied.ThesisItem Open Access SERUM ALKALINE PHOSPHATASE POLYMORPHISM IN CROSSBRED CATTLE OF KERALA(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 1995) JACOB ABRAHAM; B. NANDAKUMARANThe present investigation was undertaken to identify the the biochemical polymorphism at serum alkaline phospha- tase (SAP) loci and to study the heterogenity of SAP variation in crossbred cattle. It was also envisaged to analyse the association of SAP variation and traits of economic importance such as milk production and composition of milk. One hundred and ten animals belonging to two different crosses of local nondescript cattle viz. Crossbred Holstein Friesian (57) and Crossbred Brown Swiss (53) were typed for SAP variance by standardising Horizontal Polyacrylamide Gel Electrophoresis (PAGE). Two genotypes FS and SS were determined. The highest frequency of FS genotype was in Holstein Friesian crossbred than in Brown Swiss crossbred. The genotype FF was absent in both the crossbreds. The highest frequency of SS genotype was in Brown Swiss crossbred than in Holstein Friesian crossbreds. Two alleles namely pF and pS with two phenotypes FS and SS were identified as SAP locus. pT allele had the frequency of 0.20 and pS allele had the frequency of 0.80 in the pooled crossbreds. Both the Holstein Friesian crossbreds and Brown Swiss crossbred are in genetic equilibrium at the SAP loci. No association could be established between milk yield (305 days) and serum alkaline phosphatase level. A non significant negative correlation existed between milk fat percentage and SAP level whereas a significant positive correlation existed between milk SNF percentage and SAP level. The correlation between SAP level and milk total solids were found to be negative and non significant in Brown Swiss 1 I i / crossbreds whereas a non significant positive correlation existed between the SAP level and milk total solids in Holstein Friesian crossbreds. Animals belonging to the FS genotype are better milk producers compared to the SS genotype. For higher fat percentages, the performance of SS genotype was compara- tively better. The performance of SS genotype Ls better for producing milk with more than 8.5 percentage of SNF. FS genotype performed better for producing milk having higher percentage of total solids.ThesisItem Open Access CHARACTERISATION AND EVALUATION OF THE DWARF CATTLE OF KERALA(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 1994) C. R. GIRIJA; Sosamma lypeThe native cattle of Kerala have been treated as non descript animals always eventhough they possess some special features. The dwarf cattle often called as Vechur were very popular in Central Travancore until 35 years back. with the emergence of the crossbred population of cattle the traditionally reared local cattle have gradually suffered genetic erosion. Under this circumstance, the present work was undertaken to characterise and evaluate the germplasm of local dwarf cattle of Kerala by studying (a) the karyotype and morphology of chromosomes using G-banding (b) the population structure by means of gene frequencies of different blood proteins (c) the growth and production performance. The characterisation and the evaluation would help in finding out the genetic differences of the dwarf cattle which will help in deciding about the conservation of their germplasm as a reserve for the future. The dwarf cattle maintained under the ICAR scheme on "Conservation of germplasm of Vechur cattle of the coastal area and the dwarf cattle of the high ranges of Kerala" formed the material for the study. The characterisation and evaluation was carried out through the cytogenetic, immunogenetic and polymorphism studies as well as through the description of the growth and production traits. Karyotype analysis was carried out using peripheral blood leukocyte culture technique described by Hainan (1977) and Hainan (1989) with suitable modifications. G-banding of chromosomes were done by the method described by Thiagarajan, ^19 9!^. Blood protein polymorphism systems^ such as Haemoglobin and transferrin were studied by poly acrylamide gel electrophoresis in horizontal dimension ' y(Cahne et al. ,1977) with suitable modifications. The statistical analysis of the growth and production data were done as suggested by Snedecor and Cochran (1967). The diploid chromosome number of the dwarf cattle was found to be 60, with 29 pairs of autosomes and one pair of sex chromosomes. All the autosomes and the 'Y' chromosome were acrocentric. The X chromosome was submetacentric. The relative length of the autosomes ranged from 1.757 to 5.431 per cent. The relative length of the X and Y chromosomes were found to be 5.591 per cent and 2.875 per cent respectively. In the karyological array, the X chromosome occupied the first position. The X chromosome was biarmed and the arm ratio and centromere index obtained were 2.182 and 0.314 respectively. The karyotype and morphometric measurements resembled the finding in Bos indicus group of cattle. The G-banding pattern of chromosomes revealed 72 regions and 314 G-bands. The Y chromosome had 7 G-bands in the 'q' arm which resembled the 'q' arm of Bos taurus described in the international system for cytogenetic nomenclature of domestic animals. A B There were two haemoglobin variants Hb and Hb and three phenotypes viz. Hb^^, Hb^® and Hb®^, in the population. The heterozygocity was found to be 0.4815. The population was found to be in genetic equilibrium with respect to the Haemoglobin locus. Six transferrin phenotypes controlled by three alleles Tf^, Tf*^ and Tf^ were observed. The frequency of Tf^ (0.359) allele in the dwarf cattle was as high as the frequency of the allele reported in the zebu cattle. The absence of F H N G transferrin variants like Tf , Tf , Tf and Tf and higher frequency of Tf allele are probably indicative of the genetic isolation of the population from exotic breeds. The absence B F of Tf and Tf allele which is present in Gir, Hariana, Kankrej, Kangayam, Ongole, Red Sindhi, Sahiwal and Tharparkar also indicates that the dwarf cattle has not inherited genes from the above cattle breeds. The body weights and measurements of calves at birth studied showed that the male calves had a higher body weight (12.55 + 0.311 kg with a CV of 7.86 per cent) than female calves (10.78 + 0.404 kg with a CV of 15.02 per cent). The same trend was observed with regard to the birth body measurements also. The heart girth measurement and body weight showed a positive correlation from birth to the 24th fortnight. There is a 100 per cent increase in the birth weight by the 5th fortnight and a three-fold increase by the 10th fortnight. The average daily gain in weight for the four periods i.e., fortnights 0-6, 7-12, 13-18 and 19-24 were 0.160 + 0.011, 0.167 + 0.018, 0.212 + 0.011 and 0.139 + 0.015 respectively for female calves, and 0.188 + 0.023, 0.145 + 0.016, 0.116 ^ 0.025, 0.242 + 0.049%respectively in male calves. During the period from birth to 6th fortnight the growth rates in males and females were similar. The gain in body weight per day during the periods from 7 to 12th and 13 to 18th fortnight was comparatively less for males but the trend reversed during the period of fortnights for 19 to 24th. The average body weights of adult females and males were 126.90 + 3.56 kg (CV 16.39%) and 210 + 15.75 kg (CV 14.95%) respectively. The body measurements such as length, heart girth and height (in cms) in females were 97.5 + 1.12 (CV 5.85%), 115.60 + 1.32 (CV 5.82%) and 87.53 + 0.82 (CV 4.82%) respectively. The corresponding figures in males were 111.5 + 3.77 (CV 6.76%), 146.0 + 2.92 (CV 3.99%) and 107.5 + 1.35 (CV 2.50%) respectively. The average body weights and measurements were lesser than those reported in other Indian breeds and crossbred cattle. The total lactation milk production performance of the dwarf cattle was 471.68 + 38.72 kg (cv 45.29%) in an average lactation length of 217 + 16.50 days (CV 32.20%). The average daily yield was 2.17 + 0.11. kg (cv 29.48%). • The dwarf cattle attained a peak yield of 3.71 + 0.16 kg (cv 21.5%) in 23.23 + 1.703 days (CV 37.38%). The milk production performance eventhough was lesser than crossbreds or some recognised Indian breeds, the milk production in comparison with the body size was reasonable. Considering the morphology of the Y chromosome, the Hb as well as Tf polymorphism and their allelic frequencies, it is to be summarised that the stock of dwarf cattle of Kerala maintained at Kerala Agricultural University is gehetically isolated from the other cattle breeds of the country and world. The body size and milk production of the cow indicates its suitabMity for a farmer who requires milk just for home consumption. The study strongly confirms the necessity of conservation of the dwarf cattle of Kerala which is the smallest variety available in India and perhaps in the world itself.ThesisItem Open Access STUDY OF GENETIC DIVERSITY IN MALABARI GOATS (Capra hircus) UTILIZING BIOCHEMICAL AND IMMUNOLOGICAL MARKERS(College of Veterinary and Animal Sciences, Mannuthy., 2006) K. A. BINDU; Dr.K.C.raghavanABSTRACT Goat populations of Tanur, Thalassery and Badagara were studied for biochemical polymorphisms, immunological and microsatellite markers to investigate the similarities and differences between these populations. With regard to biochemical markers tested, polymorphism was observed only for haemoglobin, transferrin and glutathione loci. Two variants were observed for haemoglobin, Hb and Hb with a frequency of 0.987 and 0.012, respectively, suggestive of three phenotypes, viz. Hb aa, Hb ab and Hb bb, and indicating the predominance of Hb in the pooled population. Hb ^ variant was observed only in the Thalassery population (gene frequency 0.038). Two variants for transferrin (Tf^ and Tf were detected with a predominance of TC in the population. All the goats from Thalassery population belonged to Tf aa type. In the present study only two phenotypes as regards transferrin locus could be observed, (Tf AA and Tf bb) with,the notable absence of Tf ab- No polymorphism was observed for albumin, cerruloplasmin, amylase and carbonic anhydrase loci in all the animals tested. The animals were classified as low and high glutathione types based on the values obtained for blood glutathione concentration. In the pooled population, majority of the animals belonged to low GSH type (53.68 per cent). The least square analysis of glutathione concentrations showed significant variation between populations. With regard to potassium loci, all the animals in the present study belonged to low potassium type, with the mean potassium concentration of the pooled population recorded at 4.18 :L0.09 meq/1. The least square analysis of variance of potassium concentrations showed that there existed significant difference between different sub-populations. Genetic distance was calculated as described by Balakrishnan and Sanghvi (1968), using the allelic frequencies of protein polymorphic loci. Genetic distance between Tanur and Badagara was found to be 0.1249 , while that between Tanur and Thalassery was 0.6690 and between Badagara and Thalassery was 0.3351. The only possible conclusion that could be arrived at from the above studies is the existence of a relationship between these populations. Hence an attempt was made to study the different populations at molecular level, using microsatellite markers. Three markers, viz. INRA 063, HUJ 1177and ILSTS 030 were found to be polymorphic. Based on the genetic distances, it was found that Thalssery and Badagara were closely related than Tanur population.. This finding, much in agreement with biometrical traits, reiterates the close relationship between the Thalassry and Badagara populations. On an average, the goats of Thalassery and Badagara were heavier in comparison to Tanur goats, though the prolificacy remained higher in Tanur animals than the other two populations. The different sub-populations under the present study were also screened for the antibody response to SRBC. The highest concentration of antibody was observed on day seventh after primary immunization. The titre gradually reduced by the 15"^ day, reaching the lowest values on IT"^' day of post immunization. The effect of antibody response to SRBC on the 7"^, IS"*" and 2C days post immunization was not found to be significant for the occurrence of diseases like diarrhoea and pneumonia. The cutaneous response to intradermal injection to Phytahaemagluttinin - M (PHA-M) was also studied to find out the differences, if any in and between the various sub- populations under study. The values for skin thickness were maximum at 24 hours post-intradermal injection of PHA-M and were recorded as 3.24±0.05, 3.23±0.06 and 3.33±0.06 mm in Tanur, Thalassery and Badagara, respectively. The skin thickness reduced considerably after 48 hours and reached 1.61±0.02, 1.62±0.02 and 1.65±0.02 mm, respectively at 72 hours. The least square analysis of variance revealed that the values for pre and post immunization skin thickness at 0, 24, 48 and 72 hours were non significant between different sub-populations. Total protein, albumin and globulin concentrations also were estimated. The highest mean concentration for globulin was detected in Badagara population (3.28±0.22g/dl) and the lowest in Tanur (2.340 ± 0.3]g/dl). The least square analysis of variance of globulin concentration revealed significant difference between populations. Though all populations under study had all predominant physical characteristics of the Malabari breed, the Tanur population stood apart as regards the biometrical characteristics, like litter size and body weight and charecteristics perceivable at the molecular level. It could well be inferred that this population might have evolved through mixing up of the local nondescript Tanur goats with original Malabari goats. The study reiterates the need for more research activities directed at exploring the chances of conserving and developing such unique populations within a breed.ThesisItem Open Access GROWTH AND SURVIVABILITY OF GH/Msp I GENOTYPES IN MALABARI GOATS(College of Veterinary and Animal Sciences, Mannuthy., 2004) BINDU MATHEW; Dr. k.C. RaghavanGrowth hormone gene, due to its essential role in lactation and growth processes, is a perfect candidate marker associated with somatotropic axis. Selection of animals based on the growth hormone genotypes can be tntroduced in the animal husbandry sector for better production. The present investigation was undertaken to study the growth and survivability of GH/Mspl genotypes in Malabari goats at different centers of Badagara. Thalassery. Thanur as well as Malabari conservation unit. Regional Agricultural Research Station, Pilicode. DNA was isolated from 32 bucks, 241 does mated to those bucks and 297 of their progeny using phenol - chloroform extraction method. A 768-bp fragment from third exon to fifth exon containing the polymorphic Mspl site was amplified well using bovine primers, indicating species homology. The amplified product on digestion with the Mspl enzyme revealed the GWMspl {+) and (-) alleles. The percentage of incidence of (+/-) genotype was 61.76 and that of (+/+) genotype was 38.24. None of the animals typed were of the GH/Mspl (-/-) genotype. The genotype frequencies of bucks and does were consistent with the general population. In the specific heterozygous mating (+/- x +/-), 28 per cent of the progenies were (+/+) homozygotes and the rest 72 per cent were heterozygotes (+/-). Early embryonic mortality was not found to be a cause for the absence of the GH/Mspl -/- genotype as the kidding percentage in heterozygous mating were not different from other types of ma.rngs. There was no relationship between growth upto SIX months of age and G\l!Msp\ genotypes. The study conf.mrs a strong heterozygotie advantage for the GH/Mspl +/genotype and also the absenee of GH/Mspl -/- genotype in Malaban goats. Presence of duplicate copies of the growth hormone gene in goats may be a possible reason for the above results.ThesisItem Open Access MICROSATELLITE MARKER BASED CHARACTERIZATION OF INDIGENOUS PIGS OF KERALA(College of Veterinary and Animal Sciences, Mannuthy., 2006) ARIPRASATH. K; Dr. A.P. UshaThe study was undertaken to assess the genetic diversity among four indigenous pig population of Kerala using microsatellite markers. The animals were selected from various part of Kerala, population I included the conserved Angamali pigs from university farm, Mannuthy, population II consisted of animals from Koothattukualm, population III were the animals from Ollur and animals from border districts of Kerala formed the population IV. Genetic analysis was carried out using five polymorphic microsatellite markers. Blood samples were collected from 100 unrelated indigenous pigs from all four populations and DNA was isolated. The phenol-chloroform method of extraction yielded 224.35±9.86pg/5ml of blood. PGR conditions were standardized for all five selected markers namely, S0005, SOlOl, SW1026, SW2517 and S0008. The forward primer of each marker was endlabelled with 732 P-ATP as source of radio signal. The Ml3 single strand DNA was sequenced and used as a size standard. Autoradiography was employed to visualize the results. A total of eight alleles were detected in S0005 and SOlOl, five alleles in each of SW1026 and S0008, and six in SW2517. The heterozygosity varied from 0.7747 in SW2517 to as large as 0.8475 for S0005. The heterozygosity values for SOIOI, SW1026 and S0008 were 0.7774, 0.7672, and 0.7424 respectively. The PIC values ranges from 0.6974 for S0008 to 0.8291 for S0005. The PIC values for SOIOI, SW1026 and SW2517 were 0.7483, 0.7284 and 0.7381 respectively. The allele frequencies were used to estimate the Nei's standard genetic distance among the populations. The distance measure ranged from 0.5704 to 0.7161, with the highest value noticed between population II and IV and the lowest between population I and III. A dendrogram was constructed using the POPGENE version 3.2 program which grouped the population I and IV in one cluster and II and III populations in another cluster. >ThesisItem Open Access MILK PROTEIN POLYMORPHISM AND MILK COMPOSITION IN VECHUR CATTLE(COLLEGE OF VETERINARY AND ANIMAL SCIENCES Mannuthy - Thrissur, 1996) THIRUPATHY VENKATACHALAPATHY, R.; SOSAMMA IYPE; THIRUPATHY VENKATACHALAPATHY, R.The native cattle of Kerala have been evolved through several generations of natural selection against high humidity, heavy rainfall and hot climate. They have been considered as non-descrlpt animals, but there has been variety with characters distinguishing from others and known as Vechur cattle of Kerala. The Vechur cattle had their origin In Vechur near Valkom of Kottayam district of Kerala. They were very popular four decades back for their relatively higher milk production compared to other, local cows. The extremely small size of the cow, . good adaptation and high disease resistance are the traits very much favoured by the farmers. Farmers prefered Vechur cattle for ploughing In marshy lands because of the small size and light weight. The emergence of crossbreeding led to the gradual reduction In tradatlonally reared Vechur cattle. Under this circumstances, the present work was undertaken to characterise the Vechur cattle of Kerala by studying (a) certain genetic markers In milk (b) milk components like fat, total solids and solids not fat percentage (c) fat globule size and distribution and (d) Iodine value. The Vechur cows maintained under the scheme on "Conservation of germplasm of Vechur cattle" formed the material for the study. Milk proteins such as casein and whey proteins were studied by polyacrylamide gel electrophoresis in horizontal dimension (Thompson and Kiddy, 1963). The fat, total solids, solids not fat percentage of milk and iodine value of milk fat were estimated as prescribed by Indian Institution Standards. The fat globule size was measured under microscope by fitting an eye piece micrometer to the eye piece of the microscope. The statistical analysis were done as suggested by Snedecor and Cochran (1967). The influence of stage and time of milking were worked out using least squares analysis of variance as described by Harvey (19 86) . Three proteins a, f3 and k-casein were identified in casein system. At a^^-casein locus three phenotypes viz. BB, BC and CC determined by two allele B and C were observed. Frequency of CC phenotype (0.64) was highest ^ followed by BC (0.22) and BB (0.14). Frequency of B and C allele were 0.25 and 0.75. The /3-casein locus consisted three phenotypes viz. AA, AB and BB with A and B allele. The frequency of AB phenotype was highest (0.57) followed by AA (0.29) and BB (0.14). Frequency of ^ arlele was 0.5/. At k-casein locus two phenotypes AA and AB with A and B allele were identified. The frequency of AB was (0.83) and frequency of A allele was 0.59. Two whey proteins a-lactalbumin and /3- lactoglobulin were identified. At a-lactalbumin locus two phenotypes AA and AB with two allele A and B were observed. The frequency of AA and AB phenotypes were 0.57 and 0.43. The frequency of A and B allele were 0.78 and 0.22 respectively. The /3-lactoglobulin locus consisted of three phenotypes viz. AA, AB and BB with A and B allele with frequency of 0.57, 0.29 and 0.14 respectively were identified. The frequency of A allele was 0.71. The frequency of C allele at a^^-casein locus was lower than the other Indian breeds like Sahiwal, Tharparkar, Red sindhi and Rathi, but higher than exotic breeds like Jersey and Holstein and crossbred cattle of Kerala. The A variant at f3 and k-casein locus had frequency lower to other Indian breeds, exotic breeds of cattle and crossbred cattle of Kerala. The A variant at c*-lactalbumin and /3-lactoglobulin locus had much higher frequency than other Indian, exotic breeds of cattle of cattle and crossbred cattle of Kerala. B variant in Vechur cattle was lower in frequency compared to other Indian breeds. The average milk fat percentage for 1-44 weeks of lactation was 5.95 +0.12, 6.62 ± 0.13 in the morning and evening respectively and the mean milk fat percentage for morning and evening milk was 6.23 + 0.19. The mean milk fat percentage at first week was 4.05 ± 0.19, 4.65 + 0.17 and by 20 th week of lactation was 5.92 + 0.08, 6.55 ± 0.07 in the morning and evening milk respectively. The evening milk had uniformly higher fat percentage than morning milk. The fat percentage showed an increasing trend with advancing stage of lactation. The least squares means for morning and evening milk were 5.99 + 0.03 and 6.62 ± 0.03 respectively and overall mean was 6.13 ± 0.04. The mean total solids percentage of milk at first week was 12.64 ± 0.17 and 13.57 ± 0.19 in the morning and evening milk and it steadily increased to 14.75 ± 0.13 and 15.31 + 0.18 in the morning and evening milk by 20 th week of lactation. The average total solids percentage in milk from 1 -44 weeks of lactation was 14.79 ± 0.13 and 15.53 + 0.13 in the morning and evening respectively and the average for morning and evening together was 15.16 i 0.11 per cent. An increasing trend was noticed in total solids percentage as the lactation advanced. The evening milk showed uniformly higher total solids percentage compared to morning. The least squares mean was 15.02 and the adjusted mean for morning and evening was 14.85 ± 0.05 and 15.55 ± 0.05 per cent. Least squares analysis of variance showed a significant effect of time of milking and stage of lactation on milk fat and total solids percentage of milk. The average solids not fat percentage from 1 - 44 weeks of lactation was 8.84 ± 0.12 and 8.92 ± 0.14 in the morning and evening milk respectively. The average for morning and evening milk was 8.88 + 0.13 and least squares mean was 8.90. The mean solids not fat percentage of milk at first week was 8.60 + 0.24 and 8.93 ± 0.22. It was 8.83 ± 0.11 and 8.76 ± 0.15 percent at 20 th week of lactation in the morning and evening milk respectively. No trend of increase was noticed in the solids not fat percentage in relation to the stage of lactation. The least squares analysis of variance showed no significant effect of time of miling and stage of lactation on solids not fat percentage. Total solids and solids not fat did not have strong association in general. The positive correlation between total solids and fat percentage were not significant. The negative correlation between fat and solids not fat percentage were also not significant. The milk fat and total solids percentage were higher than other Indian breeds of cattle, but lower to buffalo. The solids not fat percentage of milk was similar to other Indian br-eeds of cattle. The mean size of fat globule was 3.21 and the range was 2.54 to 4.07 /.i. The mean diameter of fat globules was found to be 3 .02 + 0.05 jj, in the morning and 3.40 + 0.05 in the evening milk. The mean size of fatf globules ^timated in goat (Malabari and their crosses), crossbred cattle and Murrah buffalo were 2.60 u., 4.87 jj, and 5.81 /./ respectively. The average fat globule size was found to be decreasing as the lactation advanced. The proportion of small size fat globules were found to be increased and the larger size of fat globules decreased towards the end of lactation. No correlation was noticed between fat globules size and fat percentage. The iodine value ranged from 28.61 to 30.29 and the average iodine value was 29.60 ± 2.20. Based on available reports this appeared to be similar to buffalo, higher than goat and lower than cows. The size of fat globule (3.21 u.) was higher than the goat and lower to the other breeds of cattle and buffalo. since the milk fat has higher proportion of smaller size fat globules and saturated fatty acids, it would be therapeutically useful in malabsorption syndromes due to its easy digestability. The larger proportion of small size fat globule is associated with high phospholipid content because of greater surface area. Phospholipids are important in the development of nervous system in babies. Thus Vechur cow milk appear to be suitable for infants and even sick. Studies on more number of cows are required for confirmation of results. It can be concluded that Vechur cattle of Kerala has unique characteristics of its own and have separate identity from other breeds of cattle not only by its small size but also due to its milk protein variants, composition of milk, size of fat globules and level of saturated fatty acids.ThesisItem Open Access PORCINE IMMUNE RESPONSE AS MARKER TRAITS FOR SELECTIVE BREEDING(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2002) . RAJAN, M. R; RAGHUNANDANAN, K.V.Survivability and better performance of pigs under tropical stress have been reported to be significantly influenced by immune responses. Immune response traits under genetic control offer potential possibilities for exploited in commercial pig production. The present research project on the utilisation of porcine immune responses by estimating the magnitude of humoral and cell mediated immune responses in Desi and Large White Yorkshire attempted to evaluate the genetics of immune responses and to identify the association between the immune response traits and economic traits. The immune response traits were studied in 150 piglets aged between two to three months, 75 each belonging to Desi and Large White Yorkshire of both sexes and sired by eight sires each. The immune response traits studied were antibody response to sheep red blood cells (SRBC), delayed type hypersensitivity (DTH) to intradermal injection of PHA-M and lymphocyte transformation response to BCG. The economic traits recorded were litter size at birth, litter weight at birth, litter size at weaning, litter weight at weaning, weaning mortality and the occurrence of diarrhoea and pneumonia. Naturally occurring antibodies to SRBC could not be detected in both the breeds. Peak antibody response to SRBC was obtained at day 7 post immunisation with a mean titre of 4.830. Heritability estimates of antibody response to SRBC were 0.8969 + 0.4235. 0.9187 ± 0.4893 and 0.8174 ± 0.4893 respectively at 7*^ day, 15"' day and 21" day post immunisation. Litter size at birth and weaning had no significant influence on antibody response. Similarly, antibody response to SRBC among piglets was not influenced by the incidence of diarrhoea, pneumonia and pre-weaning mortality to a significant level. DTH responses to intradermal injection of PHA peaked at 24 hours post injection with a mean value of 3.39 mm. The mean pre injection skin thickness was 3.508 mm and 3.012 mm among Large White Yorkshire and Desi pigs respectively. This difference was found to be significant (P<0.05) and this difference was due to the significant breed difference confounding with body weight classes. The effect of breed on PHA responses at 24, 48 and 72 hours were not significant. Sex of the pig also did not influence the PHA responses significantly. The body weight classes did not influence the DTH response to PHA significantly. Sire effect was not significant on the pre injection skin thickness. But the DTH response at 24 hours was influenced by the sires in both Large White Yorkshire and Desi pigs to a highly significant level (P<0.01). At 48 and 72 hours post injection also DTH responses were influenced by sires to a significant level (P<0.05). The heritability estimates for pre injection skin thickness and DTH responses at 24, 48 and 72 hours were 0.5173 + 0.4179, 0.8136 + 0.5643, 0.6816 + 0.5187 and 0.7134 ± 0.5283 respectively. The litter size at weaning was not influenced by the initial skin thickness. DTH responses to PHA at 24, 48 and 72 hours had no significant influence on the litter size at birth and weaning. PHA responses at 24, 48 and 72 hours were not influenced significantly by the incidence of diarrhoea, pneumonia and pre weaning mortality. The analysis of lymphocyte transformation and stimulation index to BCG on zero day was around one indicating that there was not marked increase in the lymphocyte multiplication in PPD stimulated samples. The stimulation index on 15"'' day was 6.0161 in Large White Yorkshire and 6.3340 in Desi. This index further increased to 6.1070 and 6.5920 on 30'^' day and began to decline from 45"' day with a mean value of 6.0020 in Large White Yorkshire and 5.9890 in Desi pigs. The effect of breed, sex and body weight class of piglets was not found to influence the stimulation index significantly. Sire effect was not significant on the pre inoculation index while it was highly significant on 15*^. and 45*^ day in Large White Yorkshire and Desi. The estimates of heritability on 30"^ and 45*^ day stimulation index were 0.5171 ± 0.2893, 0.6289 + 0.3817 and 0.4983 ± 0.2583 respectively. Litter size at birth and weaning was not found to have any significant influence on the LT response to BCG. Correlation analysis among different immune response traits revealed that antibody response to SRBC at 7, 14 and 21'' day had highly significant positive correlation. Similarly, the correlation between PHA responses at 24, 48 and 72 hours were also highly significant and positive. PHA responses at 24 hours and LT responses at 15*^ day was also positive and significant. LT responses at 15*^ and 30th day were also significant and positive. The association between LT responses during different time intervals were always positive and significant. PHA responses were always negatively correlated with initial skin thicknesss to a significant level. Antibody response at 7, 15 and 21" day had a significantly high negative influence on the body weight at weaning. There was a significant decrease in pre weaning mortality associated with LT response at 15th day.