Assessing lameness prevalence and associated risk factors in crossbred dairy cows across diverse management environments.

BACKGROUND: A thorough understanding of lameness prevalence is essential for evaluating the impact of this condition on the dairy industry and assessing the effectiveness of preventive strategies designed to minimize its occurrence. Therefore, this cross-sectional study aimed to ascertain the prevalence of lameness and identify potential risk factors associated with lameness in Holstein Friesian crossbred cows across both commercial and smallholder dairy production systems in Bengaluru Rural District of Karnataka, India. METHODS: The research encompassed six commercial dairy farms and 139 smallholder dairy farms, involving a total of 617 Holstein Friesian crossbred cattle. On-site surveys were conducted at the farms, employing a meticulously designed questionnaire. Lameness in dairy cattle was assessed subjectively using a locomotion scoring system. Both bivariate and binary logistic regression models were employed for risk assessment, while principal components analysis (PCA) was conducted to address the high dimensionality of the data and capture the underlying structure of the explanatory variables. RESULTS: The overall lameness prevalence of 21.9% in commercial dairy farms and 4.6% in smallholder dairy farms. Various factors such as age, body weight, parity, body condition score (BCS), floor type, hock and knee injuries, animal hygiene, provision of hoof trimming, and the presence of hoof lesions were found to be significantly associated with lameness. Binary logistic regression analysis indicated that the odds of lameness in crossbred cows increased with higher parity, decreased BCS, presence of hard flooring, poor animal hygiene, and the existence of hoof lesions. These factors were identified as potential risk factors for lameness in dairy cows. Principal component analysis unveiled five components explaining 71.32% of the total variance in commercial farms and 61.21% in smallholder dairy farms. The extracted components demonstrated higher loadings of housing and management factors (such as hoof trimming and provision of footbath) and animal-level factors (including parity, age, and BCS) in relation to lameness in dairy cows. CONCLUSIONS: The findings suggest that principal component analysis effectively reduces the dimensionality of risk factors. Addressing these identified risk factors for lameness is crucial for the strategic management of lameness in dairy cows. Future research in India should investigate the effectiveness of management interventions targeted at the identified risk factors in preventing lameness in dairy cattle across diverse environments.

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.

Exploring molecular dynamic indicators associated with reproductive performance of Bos indicus cattle in blood plasma samples through data-independent acquisition mass spectrometry.

Improving reproductive performance of cattle is of paramount importance for sustainable dairy farming. Poor reproduction performance (RP) hinders the genetic improvement of important Bos indicus cattle breeds. It is well known that incorporation of molecular information along with conventional breeding method is far better than use of conventional method alone for the genetic improvement of reproductive performance traits in cattle. Therefore, the present study sought to investigate the plasma proteome of the Deoni cows in cyclical (n = 6) and pregnant (n = 6) reproductive phases with varying reproductive performance (high and low). High-throughput data independent acquisition (DIA) based proteomics was performed to understand corresponding proteome. We identified a total of 430 plasma proteins. Among cyclic cows, twenty proteins were differentially regulated in low RP as compared to high RP. BARD1 and AFP proteins were observed upregulated in cyclical cows whose upregulation reported to affect reproductive performance in cattle. Among the pregnant cows, thirty-five proteins were differentially regulated, including the downregulation of FGL2 and ZNFX1 that modulates the maternal immune response mechanism which is required for successful implantation of the embryo. Also, proteins such as AHSG, CLU and SERPINA6 were upregulated in the pregnant cows whose upregulation reported to reduced reproductive performance. The results of this study will be helpful in establishing a framework for future research on the aspect of improving reproductive performance in Bos indicus cattle breeds. SIGNIFICANCE: The Indian subcontinent is the center of domestication for Bos indicus cattle breeds and they are known for their disease resistance, heat tolerance, ability to survive in low input regime and harsh climatic conditions. In recent times, population of many important Bos indicus breeds including Deoni cattle is declining due to various factors, especially due to reproductive performance. Traditional breeding methods are not sufficient enough to understand and improve the reproductive performance traits in important Bos indicus cattle breeds. Proteomics approach is a promising technology to understand the complex biological factors which leads to poor reproductive performance in cattle. The present study utilized DIA based LC- MS/MS analysis to identify the plasma proteins associated with reproductive performance in cyclical and pregnant cows. This study if improved further, can be used to develop potential protein markers associated with reproductive performance which is useful for the selection and genetic improvement of important Bos indicus breeds.

Anti-Müllerian hormone as an endocrine biomarker of reproductive longevity and assessment of single nucleotide polymorphisms in AMH gene of Bos indicus breeds of cattle.

Anti-Müllerian hormone (AMH) is a member of the TGF-ß superfamily produced by follicular granulosa cells in women and cattle and is considered an endocrine biomarker of ovarian follicular reserve. The study examined how age and parity influence serum AMH concentration and investigated the presence of single nucleotide polymorphisms in AMH gene in Bos indicus breeds viz Malnad Gidda Amritmahal and Hallikar. All five exons of AMH gene amplified by polymerase chain reaction were subjected to sanger sequencing and identified important SNP and its effects. We observed a highly significant relationship between parity and AMH concentration in Amritmahal cattle, whereas Malnad Gidda and Hallikar breeds did not show a significant difference. We identified one SNP located in exon 5 (rs21402788) with base change A>G, a non-synonymous mutation resulting in a change in amino acid Q>R and the protein product. It is concluded that AMH level could be considered as an indicator of the ovarian reserve and productive herd life (longevity) irrespective of age/parity, especially in B. indicus breeds of cattle.

Differences in milk metabolites in Malnad Gidda (Bos indicus) cows reared under pasture-based feeding system.

The milk and milk products from cows reared under grazing system are believed to be healthier and hence have high demand compared to milk from cows reared in the non-grazing system. However, the effect of grazing on milk metabolites, specifically lipids has not been fully understood. In this study, we used acetonitrile precipitation and methanol:chloroform methods for extracting the milk metabolites followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) run to identify the different metabolites between the milk of grazing and non-grazing early lactating Malnad Gidda cows. Various carbohydrates, amino acids, nucleosides and vitamin derivatives were found to be differentially abundant in grazing cows. A total of 35 metabolites were differentially regulated (fold change above 1.5) between the two groups. Tyrosyl-threonine, histidinyl-cysteine, 1-methyladenine, L-cysteine and selenocysteine showed fold change above 3 in grazing cows. The lipid profile of milk showed a lesser difference between grazing and non-grazing cows as compared to polar metabolites. To the best of our knowledge, this is the largest inventory of milk metabolomics data of an Indian cattle (Bos indicus) breed. We believe that our study would help to emerge a field of Nutri-metabolomics and veterinary omics research.

Deep Proteome Profiling of Semen of Indian Indigenous Malnad Gidda (Bos indicus) Cattle.

Malnad Gidda is a dwarf indigenous cattle breed of India, which is known for its uniqueness of calving every year under a low input grazing system of rearing. Bulls of Malnad Gidda are known to be highly fertile even in stress conditions. However, the proteomic profiling of semen of this breed has not been investigated so far, which might provide a platform for a better understanding of its semen quality and male fertility. Therefore, we made an effort to characterize and quantify the proteome of seminal plasma and spermatozoa components of Malnad Gidda semen using a high-resolution mass spectrometry platform. We identified 2814 proteins from spermatozoa and 1974 proteins from the seminal plasma of this breed. Furthermore, >90% of proteins from each fraction were quantified using the intensity-based absolute quantification. We observed signal peptides in 33% of seminal plasma proteins, indicating their secretory nature. Gene Ontology analysis revealed their involvement in cytoskeletal assembly associated with sperm head, sperm motility, acrosome reaction, seminal plasma binding, and spermatogenesis-associated protein. An in-depth proteome profiling of semen of a unique indigenous cattle breed of India was carried out. Our findings could provide a reference for further studies on sperm functions, semen quality, and reproductive health of Bos indicus cattle. Mass spectrometry data generated in this study is deposited and publicly made available through ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD014172.

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.

Scrotal infrared digital thermography predicts effects of thermal stress on buffalo (Bubalus bubalis) semen.

The objective was to use scrotal infrared digital thermography to evaluate effects of thermal stress on semen quality of Murrah buffalo (Bubalus bubalis) breeding bulls. Ejaculates from 109 Murrah bulls maintained at three semen stations were evaluated for: ejaculate volume, sperm concentration (SCON), initial motility (IM), percent live sperm (LIVE), acrosome integrity of fresh semen (AIFS), plasma membrane integrity of fresh semen (PMIFS), head abnormalities of fresh semen (HAFS), midpiece abnormalities of fresh semen (MPAFS), tail abnormalities of fresh semen (TAFS), post-thaw motility (PTM), acrosome integrity of post-thawed semen (AIPT) and plasma membrane integrity of post-thawed semen (PMIPT). Scrotal and ocular surface temperatures were acquired during rainy, winter and summer seasons, using an FLIR i5 infrared camera. Thermographic images were analyzed with Quick Report 1.2 SP2 software and temperature data acquired. Daily mean temperature and mean relative humidity were used to determine the temperature-humidity index (THI). Environmental factors were analyzed using CORR to determine collinearity among independent variables. There was a high correlation among THI, proximal, mid and distal scrotal temperatures (r ≥ 0.73). Therefore, distal pole temperature (DPT), THI, temperature gradient (TG) and ocular temperature (OcT) were used in the analysis of variance (ANOVA) and in regression analysis of dependent variables. The THI, DPT, TG and OcT had significant influences on sperm abnormalities, acrosome and plasma membrane integrity of fresh as well as post-thawed semen. The TG had significant effects on LIVE and SCON. All semen quality parameters were predicted (regression analysis) as a function of the three independent factors. We concluded that scrotal infrared thermography was useful for assessing influences of thermal stress and environmental factors on characteristics of buffalo semen.

Luteinizing hormone beta gene polymorphism and its effect on semen quality traits and luteinizing hormone concentrations in Murrah buffalo bulls.

OBJECTIVE: Present investigation was aimed to study the Single Nucleotide Variants of the luteinizing hormone beta (LHß) gene and to analyze their association with the semen quality (fresh and post-thawed frozen semen) and luteinizing hormone (LH) concentrations in Murrah buffalo bulls. METHODS: Polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP) and Sanger sequencing method is used to study genetic variability in LHß gene. LH assay was carried out using enzyme-linked immunosorbent assay method. A fixed general linear model was used to analyze association of single nucleotide polymorphism (SNP) of LHß gene with semen quality in 109 and LH concentrations in 80 Murrah bulls. RESULTS: LHß gene was found to be polymorphic. Total six SNPs were identified in LHß gene g C356090A, g C356113T, g A356701G, g G355869A, g G356330C, and g G356606T. Single Stranded Conformational Polymorphism variants of pattern 2 of exon 1+pattern 2 of exon 2+pattern 1 of exon 3 had highly significant (p<0.01) effect on sperm concentration (million/mL), percent mass motility, acrosome integrity and membrane integrity in fresh and frozen semen whereas significant (p<0.05) effect was observed on percent live spermatozoa. SSCP variants of pattern 2 of exon 1+pattern 2 of exon 2+pattern 1 of exon 3 had highly significant (p<0.01) effect on luteinizing hormone concentrations too. CONCLUSION: The observed association between SSCP variants of LHß gene with semen quality parameters and LH concentrations indicated the possibilities of using LHß as a candidate gene for identification of markers for semen quality traits and LH concentrations in Murrah buffaloes.

Influence of season, age and management on scrotal thermal profile in Murrah bulls using scrotal infrared digital thermography.

The aim of the present study was to examine the effects of non-genetic factors on scrotal thermographic profile viz., proximal pole temperature (PPT °C), mid pole temperature (MPT °C), distal pole temperature (DPT °C) and ocular temperature (OcT) of Murrah (Bubalus bubalis) breeding bulls. A total of 109 buffalo bulls, maintained at three semen stations (SS), were monitored for scrotal surface and ocular temperatures using infrared thermography twice daily during rainy, winter and summer seasons using an FLIR i5 infrared camera and temperatures were measured. Thermograms were analysed by FLIR QuickReport v.1.2 SP2 software. Statistical analysis revealed that semen station, season, temperature humidity index (THI), housing system and timing of observations had significant (P < 0.05) effect on scrotal surface temperature (SST) and OcT. In SS-I, the PPT and MPT were significantly (P < 0.05) higher as compared to SS-II and SS-III. THI had significant (P < 0.05) effect on SST and OcT, whereas PPT (°C), MPT (°C), DPT (°C) and OcT (°C) values during high THI (>80.88; <0.05) period were higher as compared to medium THI period (70.06-80.88) and during low THI period (<70.06). Temperature gradient (TG) of the testes was significantly (P < 0.05) higher during low THI period (4.50 ± 0.06 °C) as compared to medium THI (2.38 ± 0.03 °C) and high THI (1.61 ± 0.05 °C). Season of the year had a significant effect (P < 0.05) on the SST and OcT. During the rainy season, PPT (34.50 ± 0.09 °C), MPT (33.44 ± 0.12 °C) and DPT (32.11 ± 0.15 °C) were significantly (P < 0.05) higher as compared to winter and summer seasons. Age of the bulls had non-significant effect on SST and OcT but had a marked influence on thermal profile of scrotum. It could be concluded semen station, season, temperature humidity index, housing system and timing of observations had a significant influence on scrotal surface temperature. The monitoring of scrotal surface temperature by infrared thermography was found to be useful in evaluating the effects of thermal stress on physiology and health of buffalo bulls.

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.

Peripheral plasma progesterone: a marker for determining cyclicity in yaks (Poephagus grunniens L.).

An attempt was made to determine cyclicity in yaks using plasma progesterone during the breeding and non-breeding seasons. Fifteen non-lactating yaks were used in this experiment. During the breeding season (July to November), blood samples were collected from 8 yaks at least twice weekly until estrus was observed and then at 2 days interval for 30 days. During the non-breeding season (February to March), blood samples were collected from the same number of yaks at 2-day interval for 30 days. Progesterone was determined in plasma samples by radioimmunoassay. During the breeding season, plasma progesterone at estrus was basal (< or = 0.2 ng/ml). Concentrations increased thereafter with a sharp increase during the late luteal phase, typically reaching peak levels around day 15. Concentrations then declined rapidly over the following 4 days, reaching basal levels at estrus. A high proportion (66.7%) of potential estrous periods (based on progesterone concentrations) went undetected, indicating that silent or weak estrus was a prominent problem in yak cows. During the non-breeding season, three animals were found to be cycling as determined by the patterns of plasma progesterone. Yet, behavioral symptoms of estrus were not observed in any of these yak cows. We conclude that peripheral plasma progesterone concentrations can be used to monitor cyclicity in yak cows effectively.