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ThesisItem Open Access THE STRUCTURE AND FUNCTION OF THE SHELL GLAND IN JAPANESE QUAIL UNDER DIFFERENT LEVELS OF DIETARY CALCIUM(College of Veterinary and animal Science,Mannuthy, 1994) P. T. PHILOMINA; M.G. Ramakrishna PillaiLiterature available on the physiological aspects of egg production in Japanese quail is scanty and many lacunae exist in the knowledge of various mechanisms involved in the formation of egg. Under the modern practice of intensive quail husbandry it is often found that the egg shells of Japanese quail are becoming thinner, more fragile and easily broken. Consequently, safety in transportation and marketability of these eggs are considerably reduced. Hence it was thought worthwhile to investigate some of the factors involved in egg production, more especially the egg shell formation. In the present investigation attempts were made to elucidate the mechanism of the egg shell formation with respect to the structural and functional peculiarities of the shell gland in Japanese quail (Coturnix coturnix "japonica) under different dietary levels of pre-laying and layer calcium. The study envisaged histological, histochemical peculiarities of the shell gland; the mineral (calcium and inorganic phosphorus) and enzyme (ALP and ACP) profile of the plasma and shell gland. The influence of dietary calcium on the body weight, development of the oviduct, egg production and egg shell quality was also investigated. Japanese quails attained sexual maturity early at the sixth week of age, by which time they attained a body weight of 131.8 + 0.600 g and started egg production. The birds grew at a faster rate in the early weeks, came into peak production at the 16th week of age, and the trend continued even at the 24th week of age, with a body weight of 198.033 + 0.220 g for good production. The length and weight of the oviduct increased from the sixth to 24th week of age. The variation in length and weight of the shell gland from the 16th and 24th week was marginal. Growth rate was almost marginal from the 16th week onwards. Histologically the quail shell gland showed similar structure to that of the domestic fowl except for the tunica muscularis. The inner circular muscle layer was thicker and outer longitudinal layer was thinner. As age advanced, at the 16th and 24th week of age the mucosal foldings were numerous. The inner circular muscle layer became thicker and even this layer invaded the lamina propria and the vascularity in the connective tissue core of the tunica muscularis increased. The egg quality traits such as egg weight, shell weight, shell thickness, shape index and specific gravity markedly improved as age advanced from the sixth to 16th week. From the 16th to 24th week the egg quality was lowered with an increase in egg weight, lowering of shell weight, shell thickness and specific gravity. Dietary calcium significantly influenced the body weight, egg production, shell quality, histology of the shell gland and mineral and enzyme profile of plasma. Birds exhibited optimum growth, and production at higher pre-laying and layer dietary calcium levels. Pre laying dietary calcium level of 0.7 to 0.9 per cent and layer level of 3 per cent were found to be optimum in quail ration. As far as the oviduct development is concerned, pre-laying dietary calcium had negligible influence, whereas layer dietary calcium level of 2.5 per cent was just sufficient for optimum development of oviduct/shell gland. Histologically the only difference noticed in highest (3.5%) layer dietary # calcium fed quails was the increased vascularity in their connective tissue core of the lamina propria of the shell gland. There was a positive correlation between dietary calcium (pre-laying and layer) level and plasma calcium concentration at the sixth, and 16th week of age. But at the 24th week of age only layer dietary calcium induced such a relationship. As age advanced, pre-laying dietary calcium became insignificant. In the case of plasma inorganic phosphate concentration, there existed a negative correlation to that of pre-laying and layer dietary calcium levels at all the age groups of quails. It may be emphasized in this context that the extent of the negative correlation existing between plasma inorganic phosphate and pre-laying and layer dietary calcium levels was considerably lower. The concentration of calcium and inorganic phosphorus in the plasma and shell gland were uninfluenced by the age. The shell gland concentration of calcium and inorganic phosphorus was lower compared to their plasma level. These minerals were not stored in the shell gland for shell calcification. At the time of calcification calcium was transferred from the plasma (blood) to the shell gland and then to the shell. Dietary calcium significantly influenced the plasma concentration. Plasma enzyme concentration of ALP and ACP was negatively correlated with dietary calcium (pre-laying and layer) levels, whereas the shell gland ALP concentration was uninfluenced by the dietary calcium and age and its concentration was very low compared to those of plasma ALP and shell gland ACP. Shell gland ALP was unimportant in shell formation. Histochemical localisation of the shell gland ALP was in confirmation with that of quantitative estimation. Shell gland ACP content was comparatively higher than ALP, which was also in agreement with histochemical localisation. Shell gland ACP may be involved in the transfer of calcium from the plasma to the shell through the shell gland mucosa# since its level in the shell gland was higher and influenced by dietary calcium. Variation in the plasma concentration of ALP and ACP as influenced by dietary calcium levels were related to the cyclic medullary bone formation and resorption which is a normal feature in laying birds. Variations in the dietary calcium induced changes in the concentration of plasma minerals and enzymes supported the view that dietary calcium is important for proper shell formation. Higher level of dietary calcium improved the egg shell quality, egg production and body weight. Pre-laying dietary calcium level of 0.7 to 0.9 per cent and layer level of of 3 per cent were found to be optimum in quail ration.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 INFLUENCE OF DIETARY SUPPLEMENTATION OF PROTEIN AND IODINE ON TAPIOCA TOXICITY(COLLEGE OF VETERINARY AND ANIMAL SCIENCE,MANNUTHY, 1994) ABDUL LATEEF; Ramakrishna Pillai, M GA three phase experiment was carried out to assess the level of cyanogenic glycoside (CNG) content in seven varieties of tapioca, its removal by various processing methods and its effect on metabolism in rats and kids. The role of dietary protein and iodine in modifying the deleterious effects of cyanide was investigated. Histopathological studies on the liver, pancreas and heart were also carried out in rats. The first phase of the study revealed that CNG content in terms of cyanide varied from 40.86 + 2.79 to 186.31 + 5.30 ng/g of fresh tuber v/ith the lowest concentration in 'fl-4' and higher concentrations in 'H-165' and ' Karkidakkan' . Processings like boiling and sun drying lowered the CNG content by about 50 per cent of the original. In the second phase 60 male albino Wistar rats, divided into six groups were given the following diet. Group I - Protein = 15%; Tapioca = Nil; Iodine = Nil Group II - Protein = 15%; Tapioca = 30%; Iodine = Nil Group III - Protein = 7.5%; Tapioca = 30%; Iodine = Nil Group IV - Protein = 7.5%; Tapioca v/ithout HCN = 30%; Iodine = Nil Group V - Protein = 15%; Tapioca = 30%; Iodine = 0.17 mg/kg Group VI - Protein = 22.5%; Tapioca = 30%; Iodine = 0.17 mg/kg 11 Performance of rats was evaluated by recording grov/th rate, feed consumption and feed efficiency for a period of 12 weeks. Dry matter digestibility v/as also determined over a period of 24 h. At the end of 10th, 11th and 12th week of the study the rats were sacrificed and endocrinological, biochemical and histopathological evaluation of blood/serum and tissues v/ere made. Performance of rats was poor on tapioca-based diet not supplemented v/ith iodine (Groups II and III). Rats fed on protein deficient diet in the presence of cyanide containing tapioca were the most affected (Group III) . Hov/ever, there \7as no significant variation in the performance in Groups V and VI compared to Group I when tapioca-based diet were supplemented v/ith iodine. The digestibility of dry matter in six groups of rats was almost inversely related to the quantity of feed consumed. There was significant increase in DNA and decrease in protein content of thyroid thereby increasing the DNA: protein ratio, follov/ed by extremely lov/ levels of thyroxine and hyperplasia of the thyroid in Group III. In Group II also there occurred a significant reduction in the levels of thyroxine and a resultant hypertrophy of the thyroid. Higher levels of dietary protein and iodine supplementation v/ere found to play a decisive role in Ill counteracting the inhibitory effects of cyanide and thiocyanate of tapioca origin especially on thyroid functions. The DHA: protein ratio per unit weight of pancreatic tissue v/as increased in groups fed on low levels of dietary protein (Groups III and IV). Insulin levels in different groups of rats were found to be primarily related to the levels of blood glucose. There v/as no significant difference in insulin-to-glucose ratio between the groups. The synthesis and release of insulin from the pancreas was not affected by cyanide present in the diet of tapioca fed groups. Rats showed no definite pattern of response in their blood glucose levels to cyanide contained in the diet of tapioca fed groups. There was elevation in the levels of serum total protein, cholesterol and lipid concentrations in rats fed tapioca v/ithout supplementation of iodine (Groups II and III). This may be the result of the development of hypothyroidism. However, the increase in serum total protein and lipid in Group II v;as not significant. The results indicated that the above parameters could be used as reliable biochemical indicators of thyroid activity. A significant reduction in haemoglobin concentration IV in rats of Groups II and III compared to Groups I and IV reflected the deleterious effects of cyanogens on thyroid functions as v/ell as the importance of availability of dietary protein for haemoglobin synthesis. The hepatic cells in Group II appeared slightly swollen with occasional cells showing vacuolar changes. Pancreas showed slight oedema and occasional mild degenerative changes of exocrine cells. Also there was slight interstitial oedema in the myocardium. Intensity of pathological changes in the liver, pancreas and heart v^ere more severe in rats fed on protein deficient, tapioca diet (Group III) compared to that of Group IV in the diet of v;hich tapioca was made cyanide free. The hepatic cells in Group III shov/ed marked diffused vacuolar changes with condensed or vesicular nuclei and occasional cell necrosis. Many pancreatic exocrine cells shov/ed vacuolar and degenerative changes. Interstitial oedema v/as prominent. Inspite of these pancreatic exocrine changes, it is important to note that the islets remained almost intact in all the groups studied. Myocardium showed interstitial oedema v/ith swollen sarcoplasm and fragmented sarcolerama. At the same time only moderate vacuolar changes in the liver, distention of interlobular space and interstitial oedema in myocardium were observed in Group IV. Since the diet of Groups V and VI were supplemented v/ith iodine, no significant cytological alterations could be seen in their organs. V In the third phase of the study thirty 2^5-3 months old Alpine-Malabari kids of both sexes were divided into 3 groups. They were given the following diet. Groups I Protein = 15%; Tapioca = Nil; Iodine = Nil Groups II Protein = 15%; Tapioca = 30%; Iodine = 2 mg/kg Groups III Protein = 25%; Tapioca = 30%; Iodine = 2 mg/kg The study included evaluation of the performance of kids and determination of certain endocrinological and biochemical parameters in their blood at fortnightly intervals for 3 months. There was no significant effect on the performance (body weight gain, dry matter intake and feed efficiency) of kids fed on tapioca-based diet supplemented with iodine, compared to Group I. However, the significant difference in body weight gain between tapioca fed groups (Groups II and III) was probably due to the difference in the relative availability of protein in their diet. The results of the study in kids indicated that there \/as no significant influence of cyanogenic principles present in tapioca on the thyroxin and insulin levels v/hen iodine was supplemented in the diet. Similarly the blood glucose, serum total protein, cholesterol, lipid and haemoglobin levels v/ere also not influenced throughout the study.