Digital infrared thermal imaging of udder skin surface temperature: a novel non-invasive technology to monitor calving process in Murrah buffalo (Bubalus bubalis).

Quantifiable decline in the maternal body temperature during the pre-calving offers the possibilities for predicting the calving that can improve the calving management. As infrared thermography (IRT) is a simple non-contact tool for precise measurement of surface temperature, we investigated the use of IRT to establish thermal signatures around calving in the Murrah buffalo. The IRT of eye, right lateral, left lateral and rear side of udder skin surface temperature (USST) were recorded at 6 h interval from 96 h before the expected date of calving, at the time of calving and 24 h post-calving in Murrah buffaloes (n = 28). In parallel, blood samples were collected for progesterone (P4) assay. The results revealed that the IRT of the eye, right and left lateral and rear side of USST showed a significant decrease in the temperature from 48 h pre-calving till the onset of calving with a ΔT (°C) of 0.56, 0.91, 0.70, and 0.90, respectively. Mean USST significantly declined from 48 h pre-calving with a ΔT of 0.85 °C. The residual temperature of both eye and various ROI of the udder also followed a similar and significant declining trend from 48 to 0 h of calving indicating that circadian influence on the USST was minimum. Plasma P4 concentration significantly decreased from 72 h pre-calving till calving. It is concluded that a marked reduction in the IRT of the USST at 6-12 h pre-calving would be useful in predicting the onset of calving in the Murrah buffalo.

Biological methylation: redefining the link between genotype and phenotype.

The central dogma of molecular biology is responsible for the crucial flow of genetic information from DNA to protein through the transcription and translation process. Although the sequence of DNA is constant in all organs, the difference in protein and variation in the phenotype is mainly due to the quality and quantity of tissue-specific gene expression and methylation pattern. The term methylation has been defined and redefined by various scientists in the last fifty years. There is always huge excitement around this field because the inheritance of something is beyond its DNA sequence. Advanced gene methylation studies have redefined molecular genetics and these tools are considered de novo in alleviating challenges of animal disease and production. Recent emerging evidence has shown that the impact of DNA, RNA, and protein methylation is crucial for embryonic development, cell proliferation, cell differentiation, and phenotype production. Currently, many researchers are focusing their work on methylation to understand its significant role in expression, disease-resistant traits, productivity, and longevity. The main aim of the present review is to provide an overview of DNA, RNA, and protein methylation, current research output from different sources, methodologies, factors responsible for methylation of genes, and future prospects in animal genetics.

Metabolomics-a powerful tool in livestock research.

Advancements in the Nuclear Magnetic Resonance (NMR) and Mass Spectrometry (MS) along with recent developments in omics sciences have resulted in a better understanding of molecular mechanisms and pathways associated with the physio-pathological state of the animal. Metabolomics is a post-genomics tool that deals with small molecular metabolites in a given set of time which provides clear information about the status of an organism. Recently many researchers mainly focus their research on metabolomics studies due to its valuable information in the various fields of livestock management and precision dairying. The main aim of the present review is to provide an insight into the current research output from different sources and application of metabolomics in various areas of livestock including nutri-metabolomics, disease diagnosis advancements, reproductive disorders, pharmaco-metabolomics, genomics studies, and dairy production studies. The present review would be helpful in understanding the metabolomics methodologies and use of livestock metabolomics in various areas in a brief way.

Short-term effect of foot-and-mouth disease (FMD) vaccination on the milk yield in the Deoni and crossbred cows.

Biannual vaccination of the cattle with inactivated foot-and-mouth disease (FMD) vaccine is the control strategy in endemic countries. Reduction in the milk yield is one of the main reasons for poor compliance of the cattle owners to FMD vaccination. As it can adversely affect the herd immunity, the present study aimed to quantify the losses in the milk yield post-FMD vaccination. Retrospective data on the milk yield (kg) recordings, days in milk, parity, and age at vaccination of the Deoni and crossbred cows were collected from 10 days before (-10) to 10 days after (+10) FMD vaccination (dpv). Days in milk were categorized into three stages of lactation for Deoni and crossbred cows. Age (month) was categorized into four classes. Least squares means of the milk yield were generated after adjusting for year, age, parity, and stage of lactation. Based on exploratory data analysis, the corrected milk yield records from -2 to +2 dpv for 5 years comprising 614 data points on Deoni cows (n=54) and 488 data points on crossbred cows (n=55) were used for the final analysis. Because of the correlated errors on the corrected milk yield, linear mixed model ANOVA was done by fitting dpv as fixed effect and cow as random effect, and the results revealed the effect of dpv was non-significant (P>0.05) in either breed. With respect to dpv 0, a marginal reduction of 90 g in the corrected milk yield in the Deoni cow was recorded on dpv 1, while the reduction was about 360 g on dpv 0 as compared dpv -1 in the crossbred cow. It was concluded that FMD vaccination caused a transient non-significant reduction in the milk yield in the Deoni and crossbred cows.

Characterization of antisperm antibody binding patterns in relation to sperm phenotypic attributes and field fertility in dairy bulls.

To test our hypothesis that antisperm antibodies (ASA) might alter sperm phenotypic attributes thus leading to sub-fertility/infertility in bulls, ASA were generated in crossbred male calves by immunizing with sperm two times. Cryopreserved spermatozoa from crossbred bulls (n = 24) with different field fertility ratings were incubated with ASA and different patterns of ASA immunolocalization were studied. In addition, sperm membrane integrity, acrosomal integrity and cryo-capacitation status were also assessed. Immunolocalization of sperm antigens using antisperm antibody revealed three major patterns (Acrosomal-AR, apical-AP and, acrosome and tail-AT). The proportion of ASA reactive spermatozoa was significantly (P < 0.05) higher in low- and medium-fertile bulls compared to high-fertile bulls. Among the three patterns, the proportion of spermatozoa with AR pattern was significantly (P < 0.05) higher in low- and medium-fertile bulls compared to high-fertile bulls. The proportion of membrane and acrosome intact spermatozoa was significantly (P < 0.05) higher in high-fertile bulls compared to medium- and low-fertile bulls. There were no significant differences in the proportion of cryo-capacitated spermatozoa among high-, medium- and low-fertile bulls. The relationship between ASA reactive spermatozoa and conception rates (CR) of bulls was highly (P < 0.01) significant and negative. Similarly, AR and AT pattern were also significantly (P < 0.01) and negatively related to CR of bulls. The reactivity of spermatozoa with ASA was also significantly (P < 0.01) and negatively related to the membrane and acrosome integrity of spermatozoa. It was concluded that the proportion of spermatozoa responding to ASA was higher in low-compared to high-fertile bulls and ASA localization in sperm acrosomal area was negatively related to sperm membrane and acrosomal integrity and bull fertility.

Monitoring foot surface temperature using infrared thermal imaging for assessment of hoof health status in cattle: A review.

Detection of lameness early in cows is important from the animal welfare point of view and for reducing economic losses. Currently, many studies are being conducted for assessment of hoof health status by measuring the surface temperature of skin in cattle and other animal species in different parts of the world. Infrared Thermography (IRT) is able to detect lesions of hooves associated with lameness by measuring the changes in coronary band and hoof skin surface temperature. The surface temperature of a lame limb will be increased when the hoof has lesion(s). IRT has been used as a non-invasive diagnostic tool for early detection of hoof lesions based on the temperature difference between affected and non-affected hoof and maximum foot temperature on the regions of interest. In spite of having many potential applications in cattle production, factors affecting the temperature readings in thermograms must also are considered while taking images. Standard operating procedures must be established before taking thermographs under different circumstances, by considering all the factors that affect its normal function. IRT may help in minimising the cost of veterinary services, low yield, compromised fertility and culling expenses, where lameness cannot be resolved in early stages.

Association analysis of novel polymorphisms in 2', 5'-oligoadenylate synthetase gene with reproductive traits in indigenous and cross-bred cattle of Indian Origin.

2', 5'-Oligoadenylate synthetases (OAS) are important components of an interferon-mediated antiviral pathway. No polymorphisms in exonic regions of bovine OAS1 gene have been identified and associated with reproduction traits. The objective of the study was to detect and evaluate the effects of mutations in exonic region of bovine OAS1 gene with reproduction traits in cattle. DNA samples collected from 250 individual cows of two Indian dairy breeds (Sahiwal and Frieswal) of cattle were used in the study. The genetic variants of the OAS1 gene were identified with polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and sequence analysis using seven set of primer pairs. The PCR-SSCP analysis revealed polymorphism in the fragments comprising of exon 2, exon 5 and first fragment of exon 6 while the fragments of exons 1, 3, 4 and second fragment of exon 6 were monomorphic in Sahiwal and Frieswal cattle. The mutations in the amplified region comprising of exon 2 were found to have significant association with age at first breeding and calving, service period, dry period and pregnancy rate. Significant associations were found between SNPs in the exon 5 and service and dry periods of the animal, whereas the genetic variants in the first fragment of the exon 6 showed significant association with age at first breeding and calving. To our knowledge, this study demonstrated for the first time that the polymorphisms in OAS1 gene were associated with reproductive traits and it can be chosen as a candidate gene for improvement of reproductive performance of cattle.

Promoter variants of OAS1 gene are associated with reproductive performance and incidence of normal calving in cattle.

2',5'-oligoadenylate synthetase (OAS) are important components of an interferon-mediated antiviral pathway. But no polymorphisms have yet been identified and associated with reproduction and reproduction traits in the bovine OAS1 gene. The objective of this study was to detect and evaluate the effects of mutations in the promoter region of bovine OAS1 gene with production and reproduction traits in cattle. Two single nuclear polymorphisms were identified, including A to T transversion (rs715937117:A>T) and A to G transition (rs480985443:A>G) and subsequently genotyped in Sahiwal and Frieswal populations. Three haplotypes were constructed including H1 (A-A), H2 (T-A) and H3 (T-G), among them H2 was the most abundant in the Sahiwal (0.677) and Frieswal (0.838) population. The analysis revealed that genotypes of promoter region had significant effect (P ≤ 0.05) on age at first breeding, age at first calving, service period and pregnancy rate. The animals with H1H2 diplotype had higher age at first breeding and age at first calving compared to H2H2 genotyped animals. Further logistic regression analysis also revealed that the probability of normal calving was found more in H2H2 diplotype, as the odds ratios of H1H2 and H2H3 in comparison to H2H2 were 0.516 and 0.486, respectively. H2H2 diplotyped animals also exhibited higher milk production also in terms of total milk yield, 300 day milk yield and peak yield. These results outlined that the bovine OAS1 gene can be used as candidate genes for simultaneous improvement of both production and reproductive performance of cattle.

Metabolic and immunological changes in transition dairy cows: A review.

Smooth transition from pregnancy to lactation is important for high productive and reproductive performance during later postpartum period in dairy animals. On the other hand, the poor transition often leads to huge economic loss to dairy farmers due to compromised production and reproduction. Therefore, understanding the causes and consequence of metabolic changes during the transition period is very important for postpartum health management. In this review, metabolic changes with reference to negative energy balance in transition cow and its effect on health and reproduction during the later postpartum period in dairy animals are discussed besides the role of metabolic inflammation in postpartum performance in dairy animals.

Infrared thermography: A potential noninvasive tool to monitor udder health status in dairy cows.

The animal husbandry and livestock sectors play a major role in the rural economy, especially for the small and marginal farmers. India has the largest livestock population in the world and ranks first in the milk production. Mastitis is the most common and expensive infectious disease in dairy cattle. The global economic losses per year due to mastitis amounts to USD 35 billion and for Indian dairy industry ₹6000 crores per year. Early detection of mastitis is very important to reduce the economic loss to the dairy farmers and dairy industry. Automated methods for early and reliable detection of mastitis are currently in focus under precision dairying. Skin surface temperature is an important indicator for the diagnosis of cow's illnesses and for the estimation of their physiological status. Infrared thermography (IRT) is a simple, effective, on-site, and noninvasive method that detects surface heat, which is emitted as infrared radiation and generates pictorial images without causing radiation exposure. In human and bovine medicine, IRT is used as a diagnostic tool for assessment of normal and physiological status.

Investigation of body and udder skin surface temperature differentials as an early indicator of mastitis in Holstein Friesian crossbred cows using digital infrared thermography technique.

AIM: The objective of this study was to investigate the ability of infrared thermography (IRT) technique and its interrelationship with conventional mastitis indicators for the early detection of mastitis in Holstein Friesian (HF) crossbred cows. MATERIALS AND METHODS: A total of 76 quarters of lactating HF crossbred (Bos indicus × Bos taurus) cows (n=19) were monitored for body temperature (i.e., eye temperature) and udder skin surface temperature (USST) before milking using forward-looking infrared (FLIR) i5 camera. Milk samples were collected from each quarter and screened for mastitis using Somatic Cell Count (SCC), Electrical Conductivity (EC), and California mastitis test. Thermographic images were analyzed using FLIR Quick Report 1.2 image analysis software. Data on body and USST were compiled and analyzed statistically using SPSS 16.0 and Sigmaplot 11. RESULTS: The mean±standard deviation (SD) body (37.23±0.08°C) and USST (37.22±0.04°C) of non-mastitic cow did not differ significantly; however, the mean USST of the mastitis-affected quarters were significantly higher than the body temperature and USST of unaffected quarters (p<0.001). The mean±SD USST of the subclinical mastitis (SCM) and clinical mastitis-affected quarters were 38.08±0.17 °C and 38.25±0.33 °C, respectively, which is 0.72 and 1.05 °C higher than the USST temperature of unaffected quarters. The USST was positively correlated with EC (r=0.95) and SCC (r=0.93). The receiver operating characteristic curve analysis revealed a higher sensitivity for USST in early prediction of SCM with a cut-off value of >37.61°C. CONCLUSION: It is concluded that infrared thermal imaging technique could be used as a potential noninvasive, quick cow-side diagnostic technique for screening and early detection of SCM and clinical mastitis in crossbred cows.

Babesia bigemina infection in yak (Poephagus grunniens L.): molecular detection and characterization.

Yaks contribute significantly in the Himalayan high land economy. Specific information on prevalence of babesiosis in yaks is lacking. A fast and reliable PCR assay targeting Babesia bigemina small subunit ribosomal RNA sequence (SS rRNA) was laboratory standardized for molecular detection of B. bigemina in yaks. Restriction digestion of the PCR amplified 675 bp target sequence with Vsp I confirmed the prevalent species of Babesia as B. bigemina. Nucleotide sequencing and phylogenetic analysis of PCR amplified 675 bp SS rRNA sequence revealed a close genetic relationship with other bovine isolates of B. bigemina. A PCR based survey involving 94 blood samples of yak from the National Research Centre on Yak, Dirang, Arunachal Pradesh detected infection in 5.32% of yak blood samples, which was significantly higher in comparison to microscope based detection of infection in 2.13% blood smears. This is the first report on sensitive PCR based detection of B. bigemina infection in yaks and PCR-RFLP and nucleotide sequence analysis based molecular characterization of the B. bigemina isolated from yaks.

Chromosome analysis of Murrah buffalo (Bubalus bubalis ) spermatozoa using zona-free hamster oocytes.

An interspectific in vitro fertilization system was adopted to analyse sperm chromosomes of Murrah buffalo (Bubalus bubalis ). Superovulation was induced in mature female golden hamsters (Mesocricetus auratus ) to obtain a large number of oocytes. The zona pellucidae were digested by trypsin treatment. Zona free hamster oocytes were penetrated by buffalo spermatozoa capacitated with calcium ionophore A23187. Fertilized ova were cultured in TC 199 medium supplemented with 10% fetal calf serum (FCS). Podophyllotoxin and vinblastine were used to interrupt karyogamy and tubulin polymerization, respectively. Oocytes were fixed by modified gradual fixation air drying method. Slides were stained with 2% Giemsa for 45 min. Analyzable metaphase chromosome spreads were obtained from 22.4+/-3.8% of the penetrated oocytes. Of the 70 sexed spermatozoa, 38 were X-bearing and 32 were Y-bearing spematozoa.