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ThesisItem 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 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.