Expression study of pro-inflammatory cytokines and acute phase proteins in post-partum crossbred dairy cows with subclinical endometritis

Subclinical endometritis (SCE) is one of the causes of poor reproductive performance in dairy cows. The objective of the study was to evaluate the expression of pro-inflammatory cytokines [PICs; Interleukin -1 beta (IL-1β), Interleukin -6 (IL-6), and Tumor necrosis factor- alpha (TNF-α)] and Acute phase proteins [APPs; Serum Amyloid- A (SAA) and Haptoglobin (HP)] in blood and uterine flushings of postpartum dairy cows with subclinical endometritis (SCE). Animals (n=49) were screened for SCE at 60 days postpartum (dpp) using the endometrial cytobrush cytology, and those with PMN cell percentage ≥ 18% were identified as SCE cows. The blood samples and uterine flushings from SCE (n=15) and the control group (n=15) were assessed for mRNA expressions of PICs and APPs using the Real-Time PCR method (RT-PCR). The PICs and APPs except TNF-α were non-significant (p>0.05) in blood samples whereas a significant difference (p<0.05) was found in uterine flushings. The PICs and APPs were significantly up-regulated in the uterine endometrium indicating that collection of uterine flushings at 60 dpp could be used as a diagnostic method for evaluating gene expressions of inflammatory mediators in dairy cows with SCE

Electrocardiographic changes associated with hyperkalaemia in domestic cats

Hyperkalaemia is a life-threatening electrolyte imbalance because it affects cardiac conduction and can lead to fatal arrhythmias if left untreated. The present study describes the occurrence of hyperkalaemia in cats and the electrocardiographic changes associated with this electrolyte imbalance. Hyperkalaemia was identified in 83.33 per cent of the study group subjects. Acute kidney injury and obstructive uropathy were the main clinical conditions associated with it. Electrocardiographic findings in hyperkalaemia in different cats under study included peaked T waves in lead II and the precordial lead CV6LL, atrial standstill and sino-ventricular rhythm, normal sinus rhythm, ventricular tachycardia, first-degree atrio-ventricular block, bradycardia, sinus tachycardia, and atrio-ventricular dissociation. Electrocardiography should always be performed in cases suspected of electrolyte imbalances, particularly hyperkalaemia, so as to identify any fatal arrhythmias and initiate treatment at the earliest

An overview on single nucleotide polymorphism studies in mastitis research

Mastitis is an inflammatory condition of the mammary gland caused by microorganisms as diverse as bacteria, viruses, mycoplasma, yeasts and algae. Mastitis is an economically devastating disease mainly affecting the crossbred cattle in India. Control strategies against mastitis includes antibiotic therapy, vaccination, improvements in dairy cattle husbandry, farm and feeding management etc. but has met with little success.. Mastitis tolerance/susceptibility is difficult to measure directly and hence milk somatic cell count (SCC) or milk somatic cell score (SCS) is used as an indicator trait for mastitis as both traits are highly positively correlated. Single nucleotide polymorphism (SNP) marker is a single base change in a DNA sequence at a given position. SNP markers are the most preferred genetic markers nowadays. Currently most researches worldwide have been targeting molecular high density SNP markers that are linked to mastitis tolerance in an attempt to incorporate to understand the genetics of host resistance to mastitis and this knowledge will be helpful in formulating breeding programmes in an attempt to control mastitis. This article reviews various SNPs which are reported to be significantly associated with mastitis tolerance/susceptibility

Mapping local patterns of childhood overweight and wasting in low- and middle-income countries between 2000 and 2017

A double burden of malnutrition occurs when individuals, household members or communities experience both undernutrition and overweight. Here, we show geospatial estimates of overweight and wasting prevalence among children under 5 years of age in 105 low- and middle-income countries (LMICs) from 2000 to 2017 and aggregate these to policy-relevant administrative units. Wasting decreased overall across LMICs between 2000 and 2017, from 8.4% (62.3 (55.1–70.8) million) to 6.4% (58.3 (47.6–70.7) million), but is predicted to remain above the World Health Organization’s Global Nutrition Target of <5% in over half of LMICs by 2025. Prevalence of overweight increased from 5.2% (30 (22.8–38.5) million) in 2000 to 6.0% (55.5 (44.8–67.9) million) children aged under 5 years in 2017. Areas most affected by double burden of malnutrition were located in Indonesia, Thailand, southeastern China, Botswana, Cameroon and central Nigeria. Our estimates provide a new perspective to researchers, policy makers and public health agencies in their efforts to address this global childhood syndemic

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Not AvailableViperin, also known as radical S-adenosyl methionine domain-containing protein (RSAD2) is a multifunctional interferon-stimulated gene (ISG) that is activated during the viral infections. Viperin belongs to S-adenosyl methionine (SAM) superfamily of enzymes known to catalyze radical-mediated reactions and viperin inhibits a wide range of DNA and RNA viruses through its broad range of activity. The present study reports cloning and expression of bovine viperin in a bacterial expression system. PCR-based site-directed mutagenesis was carried out for deletion of N-terminal 1–70 amino acid containing amphipathic helix of viperin that interferes in protein expression and purification. The resultant truncated viperin protein was expressed in Escherichia coli, BL-21(DE3) competent cells and purified using nickel charged affinity column. The truncated 54 kDa protein was confirmed by western blot using human RSAD2 as a probe. Further, in house, hyperimmune serum was raised against the truncated viperin in the rabbit and the reactivity was confirmed by western blot using mammalian expression vector construct of viperin transfected in Baby Hamster kidney (BHK) cells and in MDBK cells infected with Foot and Mouth disease Asia I virus.Not Availabl

Veterinary World, EISSN: 2231-0916 Available at www.veterinaryworld.org/Vol.7/June-2014/11.pdf REVIEW ARTICLE Open Access An overview on single nucleotide polymorphism studies in mastitis research

overview on single nucleotide polymorphism studies in mastitis research, Veterinary World 7(6): 416-421

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Not AvailableDespite the fact that macrophages link the innate and adaptive arms of immunity, it’s role in the early infection of foot and mouth disease virus (FMDV) is largely unknown. Recently, depletion of macrophages in vivo after vaccination has shown to drastically diminish the protection against FMDV challenge in mouse model. Even the ability of macrophages to reduce or resist FMDV infection is not known hitherto. Therefore, we examined the replication ability of FMDV in mice peritoneal macrophages and the responsiveness in terms of macrophage polarization and cytokine production. Negative strand specific RT-PCR indicated replication of FMDV RNA in macrophages. Absolute quantitation of FMDV transcripts, immunofluorescence studies and titre of the infectious progeny virus revealed that replication peaked at 12 hpi and significantly declined by 18 hpi indicating non-progressive replication in the infected macrophages. Further, significant up regulation of inducible nitric oxide synthase by 8 –12 hpi and increase of M1 specific CD11c + cells by 42.6 % after infection showed that FMDV induce M1 polarization. A significant up regulation of TNFα and IL12 transcripts at 8 hpi supported that M1 macrophages were functional. Further, we studied the expression of Type I to III interferons (IFN) and other antiviral molecules. The results indicate a marked up regulation of Type I IFNα and β by 9.2 and 11.2 fold, respectively at 8 hpi. Of the four IFN stimulated genes (ISG), viperin showed a significant up regulation by 286-fold at 12 hpi in the mice macrophages. In conclusion, the results suggest that replication of FMDV in mice peritoneal macrophages is non-progressive with up regulation of Type I IFN and ISGs. Further, FMDV induces M1 polarization in murine peritoneal macrophages.Not Availabl

Role of microRNA, bta-miR-375 in Immune Sturdiness of Vechur: The Native Cattle Breed of Kerala, India #

In the present study, next generation sequencing was employed to identify and explore the differential expression profiles of microRNAs (miRNAs) in peripheral blood mononuclear cells (PBMCs) of crossbred (B. taurus x B. indicus) and Vechur (B. indicus) cattle in response to the bacterial endotoxin-lipopolysaccharide (LPS). The PBMCs from adult apparently healthy female crossbred cows and Vechur cattle, a native cattle breed of Kerala, India were stimulated with 10 μg/mL of LPS for 6 h. Among the differentially expressed miRNAs, the expression of 13 miRNAs showed statistically significant up regulation while, significant decrease in the expression of 15 miRNAs was noticed in LPS treated PBMCs of Vechur cattle compared to crossbred cows. The expression profiling of miRNA, bta-miR-375, expression of which was found to be significantly down regulated in LPS treated PBMCs of Vechur cattle with respect to crossbred cattle by the NGS studies, is presented in the present manuscript. The decrease in expression of bta-miR-375 noticed by NGS was in accordance with the results of quantitative real time PCR assay. Functional gene enrichment analysis and pathway analysis revealed significant enrichment of predicted targets of bta-miR-375 in many immune related and cell signalling mechanisms. In addition, over representation of targets of bta-miR-375 was also noticed in pathogenesis of many of the bovine diseases. The study could also identify differences in the expression of cytokines, viz. Tumour Necrosis Factor Alpha (TNFα), Interleukin 4 (IL-4) and Interferon-γ (IFNγ) between LPS treated and untreated PBMCs of crossbred and Vechur cattle

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Not AvailableFoot and mouth disease virus (FMDV) causes an economically devastating, contagious viral disease in cloven-hoofed animals. Macrophages are potent mediators of innate immune responses against viruses. The macrophage-FMDV interaction is not studied in detail, especially the innate immune responses to the virus. Here, we investigated the effects of experimental infection of FMDV on bovine monocyte-derived macrophages (MDM). The detection of negative-strand FMDV RNA by strand specific RT-PCR and demonstration of viral antigen by immunofluorescence indicated the replication of FMDV RNA and synthesis of viral proteins, respectively in the infected MDM. However, replication kinetic experiments revealed that the copy number of FMDV transcripts and virus titration peaked at 12 hpi with subsequent reduction, indicating non-progressive replication. FMDV infection upregulated the expression of RIG1 and MDA5 in MDM, suggests that the FMDV RNA are sensed by RLRs. In addition, nearly 75% of the infected MDM differentiated into M1 phenotype, characterized by the increased expression of M1-specific markers; CD80, iNOS and proinflammatory cytokines such as TNFα and IL12. The infection induced the expression of Type I IFNs, which coincided with the decline in the viral RNA and progeny virus after 12 hpi. In addition, infected MDM abundantly expressed ISGs such as PKR, OAS1, Mx1 and viperin against FMDV infection at 12 hpi. In conclusion, FMDV underwent a non-progressive replication in the bovine MDM that might be due to induction of M1 polarization and upregulation of antiviral genes.ICAR-IVR