SNP Analysis of TLR4 Promoter and Its Transcriptional Factor Binding Profile in Relevance to Bovine Subclinical Mastitis.

Bovine mastitis is a complex infectious disease that develops in the mammary gland, predominantly caused by a bacterial infection of mammary tissue. Genetic variability of mastitis is well established and depends upon different quantitative trait loci (QTL) related to mastitis resistance or susceptibility. The susceptibility is often attributed to single-nucleotide polymorphisms (SNPs) in the variable cow breed genomes. Several global investigative attempts have resulted in studies mapping mastitis to the variations in the relevant genes. Reports have been attributed to dramatic genetic expression changes in Toll-Like Receptor 4 (TLR4) genes in mastitis-positive cows. However, the mechanism behind this variable genetic expression of TLR4 genes has been studied poorly. The present study aims to investigate SCM through various screening tests like somatic cell count (SCC), electric conductivity (EC), pH, and California mastitis test (CMT) in milk samples. This study also aims to investigate possible mechanisms behind this variable expression of TLR4 by comparative SNP evaluation and transcriptional factor profile mining. So that the important genetic mutations and effects thereof can be exploited in selecting specific breeds with higher mastitis resistance and milk yield. Seventy Holstein Frisian (HF) crossbred dairy cows were selected in the present study. The animals were screened based on various diagnostic tests (SCC, pH, EC, and CMT). Blood samples (5 mL) were collected for extraction of DNA followed by amplification of PPR1 and PPR2 of the promoter region and 5'UTR of the bovine TLR4 gene using specific primers. Sanger's enzymatic DNA sequencing technique sequenced the amplified PCR products. Further, the identification of SNPs was done through various bioinformatic tools used in this study. The findings of the present study revealed that CMT, EC, pH, and SCC could be used for the early detection of subclinical mastitis. In the present study, a significant increase in the EC, pH, and SCC in milk samples of animals affected with SCM was found in comparison to the healthy animals. The present study also revealed 16 SNPs falling in TLR4 promoter and 5' untranslated region (5'UTR) sequences in mastitis-positive genotypes compared to reference genomes. The study also investigates the potential transcriptional factor program deployed in response to variable mastitis development resistance. In the present study, the allelic and genotype frequencies of all SNP variants in the three regions viz., PPR1, PPR2, and 5'UTR, were the same indicating the absence of heterozygous condition at the respective loci. The present study has wide applicability for researchers developing mastitis-resistant breeding programs and the data generated may aid in the selection of better genetic breeds. The transcription factor binding profiles can serve as concrete leads about the studies on bovine mastitis at the molecular level and may also aid global research groups working on transcription factor (TF)-based molecular pathology of mastitis.

Type 2 abomasal ulceration in cattle and buffalo: energy and lipid metabolic status, clinico-biochemical parameters, treatment, and prognostic indicators.

Twenty nine animals (22 cattle and 7 buffaloes) having type 2 abomasal ulcer (AU2) were studied in this prospective investigation, along with a control group (n = 20). Signalment, history, clinical signs, and laboratory alterations were determined. Blood, rumen fluid, abomasal fluid, and fecal samples were collected at the time of presentation. Depressed behavior, anorexia, dehydration, melena, sparse feces, and mushy rumen with substantially reduced or nonexistent motility, tachycardia, and tachypnea were common clinical findings. Colic was observed in 58% of animals. While the neutrophil count was significantly greater, the mean of lymphocytes, hemoglobin, erythrocyte count, and hematocrit were all lower (p < 0.05) than the corresponding control values. In comparison to the control values, the levels of BHBA, NEFA, lactate, bilirubin, BUN, creatinine, LDH, CK, GGT, ALP, and AST were significantly higher but levels of proteins, cholesterol, Na, K, Cl, and calcium were lower (p < 0.05). In comparison to the control values, the other hemato-biochemical parameters did not differ substantially (p < 0.05). The rumen and abomasal pH were within the reference range while rumen chloride concentration was increased in majority of animals. Higher percentage (p < 0.05) of non-survivors showed left shift and toxic alterations in neutrophils than the survivors. As compared to the survivors, the non-survivors had higher (p < 0.05) BHBA, triglycerides, total bilirubin, AST, ALP, LDH, and lactate, and lower (p < 0.05) fibrinogen and calcium. The response to medical treatment was fair, long time survival rate was good, and there was no recurrence. The follow-up period was 28 months. After treatment, both the current and the subsequent lactation's milk production were unaffected.

Association of vitamin D status with redox balance and insulin resistance and its predicting ability for subclinical pregnancy toxemia in pregnant sheep.

The study aimed to evaluate the role of vitamin D on redox balance, insulin resistance and its predicting value for subclinical pregnancy toxemia (SPT) in pregnant ewes. At four weeks pre-lambing, fifteen healthy pregnant ewes were divided into two groups, ewes with sufficient vitamin D (25-hydroxy-vitamin D (25VitD) (SVD, n = 9) and ewes with insufficient 25VitD (ISVD, n = 6). Blood samples were collected at 4 weeks pre-lambing using modified frequently sampled intravenous glucose tolerance test for the estimation of various metabolites. The baseline glucose, insulin, non-esterified fatty acid (NEFA), fructosamine, beta-hydroxy butyric acid (ß-BHA), calcium, phosphorus concentration and total oxidant status (TOS) did not differ significantly between the two groups, however, total antioxidant capacity (TAC) was significantly (p = 0.031) low in ISVD ewes. Area under the curve for glucose, insulin, elimination rate of glucose and peak insulin also did not differ significantly between the two groups. Correlation analysis revealed, positive association of 25VitD with fructosamine, calcium and TAC, and negative correlation with NEFA and TOS. Subsequent blood sampling at 2 weeks pre-lambing and at lambing showed significant difference in NEFA (p = 0.001), ß-HBA (p = 0.001), and fructosamine(p = 0.012) between the two groups. A significant time x group interaction was observed in NEFA (p = 0.019), ß-HBA (p = 0.031), and fructosamine (p = 0.026) concentration. The NEFA concentrations were increased and fructosamine decreased at 2 weeks pre-lambing and at lambing along with significantly increased ß-HBA at 2 weeks pre-lambing in ISVD compared to SVD. Taking 0.8 mmol/L ß-HBA as the cut off limit for SPT, ISVD ewes had higher odds of developing SPT two weeks prior to lambing (OD 16.00; p = 0.042) and at lambing (OD 10; p = 0.077). This study concludes that 25VitD significantly influence redox balance and energy profile and serves as a valuable predictor for SPT in pregnant sheep.

Clinico Hemato Biochemical Findings, Clinical Management, and Production Performance of Bovines with Late Pregnancy Indigestion (Type IV Vagal Indigestion).

This prospective study was conducted on 15 animals (eight buffaloes and seven cows), diagnosed with late pregnancy indigestion. Ten buffaloes and 10 cows served as the control group. The animals were in advanced pregnancy and had partial or complete anorexia, reduced water intake, loss of defecation or scanty faecal output, and mild to moderate dehydration. Heart and respiration rates were increased and rumen motility was reduced. Five animals had persistent tympany and moderate distension of left abdomen, and two animals each had bilateral abdominal distension and papple shaped abdomen. Neutrophil and lymphocyte counts were significantly higher and lower than the control values. Total bilirubin, AST, total protein, globulin, BUN, glucose, and lactate were significantly higher, and chloride and calcium were significantly lower than the control values. Levels of ALP, GGT, albumin, creatinine, cholesterol, triglyceride, fibrinogen, fibrinogen ratio, sodium, potassium, phosphorus, and magnesium did not differ significantly from the control values. Rumen chloride concentration was higher than the reference range. Majority of animals were managed symptomatically until parturition. There was no effect on fetal survival or milk yield in current and subsequent lactation. So, late pregnancy indigestion causes clinical and hemato biochemical alterations which require special consideration when treating diseased animals.

Claw lesions causing clinical lameness in lactating holstein frisian crossbred cows.

The objective of this study was to identify claw lesions causing clinical lameness in lactating Holstein Frisian (HF) crossbred cows in dairy cattle. Seventy dairy farmers were interviewed at the monthly meetings of Progressive Dairy Farmers Association of Ludhiana, Punjab, India. Ten dairy farms were randomly selected as per probability proportional to size and a total of 450 lactating HF crossbred cows were taken into the study. All the lactating cows were scored for locomotion and rear leg view index. Trimming was done in all the clinically lame animals (animals with locomotion scores 2 and 3) and equal number of animals selected randomly from those with locomotion scores 0 and 1. Various claw lesions were evaluated in both the groups. There was a significant relationship between locomotion score and rear leg view index to identify lameness. Sole ulcers and white line fissures were the lesions responsible for clinical lameness. Other lesions did not cause clinical lameness but increased the asymmetry in lactating HF crossbred cows. Both locomotion score and rear leg view index could be reliably used to identify clinical lameness in lactating cattle.