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The present study aimed to generate antibodies against predicted B cell epitopic peptides encoding bAMH for developing different ELISA models. Sandwich ELISA was determined to be an excellent technique for assessing bAMH in bovine plasma based on sensitivity tests. The assay's specificity, sensitivity, inter- and intra-assay CV, recovery %, Lower limit of quantification (LLOQ), and Upper limit of quantification (ULOQ) were determined. The test was selective since it did not bind to AMH-related growth and differentiation factors (LH and FSH) or non-related components (BSA, progesterone). The intra-assay CV was 5.67%, 3.12%, 4.94%, 3.61% and 4.27% for 72.44, 183.11, 368.24, 522.24 and 732.25 pg/ml AMH levels, respectively. At the same time, the inter-assay CV was 8.77%, 7.87%, 4.53%, 5.76% and 6.70% for 79.30, 161.27, 356.30, 569.33 and 798.19 pg/ml AMH levels, respectively. The average (Mean ± SEM) recovery percentages were 88-100%. LLOQ was 5 pg/ml and ULOQ at 50 µg/ml (CV < 20%). In conclusion, we developed a new highly sensitive ELISA against bAMH using epitope specific antibodies.
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OBJECTIVE: Iris pattern recognition system is well developed and practiced in human, however, there is a scarcity of information on application of iris recognition system in animals at the field conditions where the major challenge is to capture a high-quality iris image from a constantly moving non-cooperative animal even when restrained properly. The aim of the study was to validate and identify Black Bengal goat biometrically to improve animal management in its traceability system. METHODS: Forty-nine healthy, disease free, 3 months±6 days old female Black Bengal goats were randomly selected at the farmer's field. Eye images were captured from the left eye of an individual goat at 3, 6, 9, and 12 months of age using a specialized camera made for human iris scanning. iGoat software was used for matching the same individual goats at 3, 6, 9, and 12 months of ages. Resnet152V2 deep learning algorithm was further applied on same image sets to predict matching percentages using only captured eye images without extracting their iris features. RESULTS: The matching threshold computed within and between goats was 55%. The accuracies of template matching of goats at 3, 6, 9, and 12 months of ages were recorded as 81.63%, 90.24%, 44.44%, and 16.66%, respectively. As the accuracies of matching the goats at 9 and 12 months of ages were low and below the minimum threshold matching percentage, this process of iris pattern matching was not acceptable. The validation accuracies of resnet152V2 deep learning model were found 82.49%, 92.68%, 77.17%, and 87.76% for identification of goat at 3, 6, 9, and 12 months of ages, respectively after training the model. CONCLUSION: This study strongly supported that deep learning method using eye images could be used as a signature for biometric identification of an individual goat.
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The Anti-Müllerian Hormone (AMH) is a member of the transforming growth factor beta (TGF-ß) superfamily, playing a significant role in cell proliferation, differentiation and apoptosis. In females, AMH is secreted throughout their reproductive life span from ovaries, whereas in males it is secreted by gonadal cells at a very early stage of testicular development. AMH is a promising marker of ovarian reserve in women and can be used to measure the female reproductive lifespan. In the present study, we cloned and sequenced the GC rich AMH gene from Indian riverine buffalo (Bubalus bubalis) and goat (Capra hircus). Obtained sequences were compared to the AMH sequences of other mammals, and corresponding amino acid sequences revealed that the caprine and bovine AMH sequences are more closely related to each other than to those of other mammals. Furthermore, we analyzed the chromosomal localization of AMH genes in mammalian species to understand potential syntenic relationship. The AMH gene is localized between the sequences for the SF3A and JSRP1 genes and maintains this precise location in relation to other nearby genes. The dN/dS ratio of AMH gene did not indicate any pressure for either positive or negative selection; thus, the physiological function of the AMH gene in the reproduction of these two ruminant species remains very vital. Similar to other mammals, the AMH gene may be an important indicator for regulating female reproductive biology function in bovine, cetacean, caprine, and camelidae.
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AIM: Dairy cattle health monitoring program becomes vital for detecting the febrile conditions to prevent the outbreak of the animal diseases as well as ensuring the fitness of the animals that are directly affecting the health of the consumers. The aim of this study was to validate real-time rectal temperature (RT) data of radio frequency based digital (RFD) thermometer with RT data of mercury bulb (MB) thermometer in dairy cattle. MATERIALS AND METHODS: Two experiments were conducted. In experiment I, six female Jersey crossbred cattle with a mean (±standard error of the mean) body weight of 534.83±13.90 kg at the age of 12±0.52 years were used to record RT for 2 h on empty stomach and 2 h after feeding at 0, 30, 60, 90, and 120 min using a RFD thermometer as well as a MB thermometer. In experiment II, six female Jersey crossbred cattle were further used to record RT for 2 h before exercise and 2 h after exercise at 0, 30, 60, 90, and 120 min. Two-way repeated measures analysis of variance with post hoc comparisons by Bonferroni test was done. RESULTS: Real-time RT data recorded by RFD thermometer as well as MB thermometer did not differ (p>0.05) before and after feeding/exercise. An increase (p<0.05) in RT after feeding/exercise in experimental crossbred cattle was recorded by both RFD thermometer and MB thermometer. CONCLUSION: The results obtained in the present study suggest that the body temperature recordings from RFD thermometer would be acceptable and thus RFD thermometer could work well for monitoring real-time RT in cattle.
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AIM: The present study was conducted to explore the effect of supplemental heat on mortality rate, growth performance, and blood biochemical profiles of indigenous Ghungroo piglets in sub-tropical cold and humid climatic conditions of Tripura, a state of the north eastern hill (NEH) region of India. MATERIALS AND METHODS: The experiment was conducted on 38 indigenous Ghungroo piglets from birth up to 60 days of age. Among the 38 piglets, 19 piglets were provided with supplemental heat ranging between 17.0°C and 21.1°C for the period of the first 30 days and thereafter between 24.1°C and 29.9°C for the next 30 days. The other 19 piglets were exposed to natural environmental minimum temperatures ranging between 7.2°C and 15.0°C during the first 30 days and then between 18.5°C and 25.5°C for the next 30 days. RESULTS: The supplemental heat resulted in 10.6% reduction of piglet mortality from the 2(nd) till the 7(th) day of age. These beneficial effects could be related with the lower (p<0.05) plasma glutamate pyruvate transaminase (GPT) and cortisol levels and higher (p<0.05) plasma alkaline phosphatase (AP) concentrations in heat supplemented group compared to control group. Plasma AP, GPT, glucose, triiodothyronine, and luteinizing hormone concentrations decreased (p<0.05) gradually with the advancement of age in both control and supplemental heat treated piglets. CONCLUSION: Supplemental heat could be beneficial since it is related to a reduction of piglet mortality during the first week of life under farm management system in the sub-tropical climate of NEH region of India.
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The aim of this study was to determine if single nucleotide polymorphisms (SNPs) in the leptin receptor (LEPR) gene associated with delayed onset of puberty are associated with changes in other reproductive traits in adult ewes. The ovulation rate of ewes homozygous for the SNPs was ~15% lower (PPLEPR SNPs than their wild-type or heterozygous contemporaries. Partial failure of multiple ovulations was also increased (PLEPR had on average 0.2 fewer lambs at mid-pregnancy and at birth compared with the wild-type or heterozygous ewes (PLEPR were strongly associated with poorer reproductive performance in Davisdale ewes, which is likely to be linked to both a reduced number of ova available for fertilisation and an increased number of ewes failing to become pregnant. Increased partial failure of multiple ovulations in ewes with high ovulation rates (i.e. 3 or greater) may also contribute to the poor reproductive performance.
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Data on age and body weight at breeding, parity, previous litter size, days open and some descriptive body linear traits from 389 meat-type, prolific Black Bengal goats in Tripura State of India, were collected for 3 and 1/2 years (2007 to 2010) and analyzed using logistic regression model. The objectives of the study were i) to evaluate the effect of age and body weight at breeding, parity, previous litter size and days open on litter size of does; and ii) to investigate if body linear type traits influenced litter size in meat-type, prolific goats. The incidence of 68.39% multiple births with a prolificacy rate of 175.07% was recorded. Higher age (>2.69 year), higher parity order (>2.31), more body weight at breeding (>20.5 kg) and larger previous litter size (>1.65) showed an increase likelihood of multiple litter size when compared to single litter size. There was a strong, positive relationship between litter size and various body linear type traits like neck length (>22.78 cm), body length (>54.86 cm), withers height (>48.85 cm), croup height (>50.67 cm), distance between tuber coxae bones (>11.38 cm) and distance between tuber ischii bones (>4.56 cm) for discriminating the goats bearing multiple fetuses from those bearing a single fetus.
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Attainment of puberty is a key developmental event influenced by genetic and environmental factors. In examining age at attainment of puberty, we observed closely related rams from the Davisdale line whose daughters differed in age at which they attained puberty. A candidate gene approach was used to identify mutations that may underlie these observed differences. Four rams with divergent phenotypes for their daughter's age at onset of puberty were selected for whole-genome sequencing. The coding regions of genes with known roles in regulating reproductive function were searched for single-nucleotide polymorphisms (SNPs) that altered the amino acid sequence of the protein. Of interest were three SNPs in the leptin receptor gene (LEPR). A Sequenom assay was developed to determine the genotype of these SNPs in daughters of 17 sons of a founding sire. A higher percentage of ewe lambs homozygous for the LEPR mutations failed to undergo puberty before 1 yr of age, and those that did undergo puberty during the first breeding season on average were approximately 17 days older than homozygous wild-type ewes. Heterozygous ewes were intermediate for both measurements. Given the predicted change in protein function produced by the mutation in LEPR and the strong associations between the genotype and onset of puberty phenotypes, we propose that this mutation in LEPR underlies the observed difference in age at onset of puberty in the Davisdale line. Furthermore, these animals will likely provide a useful model to better understand the role of leptin in the regulation of puberty.
Assuntos
Estudos de Associação Genética/veterinária , Polimorfismo de Nucleotídeo Único , Receptores para Leptina/genética , Maturidade Sexual/genética , Ovinos/crescimento & desenvolvimento , Ovinos/genética , Fatores Etários , Animais , Sequência de Bases , Peso Corporal/genética , Análise Mutacional de DNA/veterinária , Feminino , Masculino , Fenótipo , Receptores para Leptina/metabolismoRESUMO
Identifying prolificacy potential and determination of fetal number during pregnancy for proper care and management of the pregnant goats bearing multiple fetuses and achieving the benefits out of multiple births are essential for sustainable goat farming. Our objectives were (1) to examine prolificacy potential in goats by using pituitary response to gonadotrophin releasing hormone (GnRH) challenge test, (2) to investigate hormonal profiles for the prediction of fetal number in pregnant goats and (3) to find out the most reliable timing of blood sampling for discriminating prolificacy trait and differentiating the goats bearing single, twin and triplet fetuses. In first experiment (GnRH challenge test), plasma FSH concentrations were significantly higher (P<0.01) among the goats belonging to triplet vs. twin vs. single kidding size groups after GnRH administration. Multivariate stepwise discriminant function analysis recognized that one blood sampling at 220min after GnRH administration can be used to distinguish prolificacy potential in goats. In second experiment, plasma progesterone levels were significantly higher (P<0.01) in goats bearing triplet vs. twin vs. single fetus between day 84 and 21 prior to parturition. Plasma estrone sulphate concentrations were found to be higher (P<0.05) in does bearing multiple fetuses than the does bearing single fetus between day 126 and 28 prior to parturition. A single blood sampling at day 63 prior to parturition was the most probable suitable time for discriminating kidding size by using plasma progesterone as marker.