Screening of InDel variants in PRDM6, myostatin and IGF2BP1 genes and association analysis with body measurement traits in Malabari and Attappadi black goats.

An insertion/deletion (InDel) polymorphism study of PR/SET domain family 6 (PRDM6), myostatin (MSTN) and insulin-like growth factor 2 mRNA binding protein 1 (IGF2BP1) genes was conducted in Malabari and Attappady black goats. An association study of identified InDels and body measurement traits was also performed. Body measurements included body length, chest diameter, chest depth, canon circumference, hip width, and hip height at the hip cross. The body trunk index, the body length index, the canon circumference index, and the chest width index were calculated. The Hardy-Weinberg equilibrium (HWE) was tested using a Chi-square test. The observed heterozygosity (Ho), expected heterozygosity (He), and polymorphism information content (PIC) were calculated. A significant difference in body measurements was found across breeds, ages, and breed x age interactions. The PRDM6 InDel was also associated with body measurement traits, such as body height, canon circumference and canon circumference index. In both Malabari and Attappadi black MSTN and PRDM6 InDels were in a state of HWE, while IGF2BP1 InDels were not. Indel markers found in the present study may be used for marker-assisted selection of growth traits among goats.

Phylogenetic analysis of Babesia gibsoni isolates of south India using apical membrane antigen, 50 kDa surface antigen, and 70 kDa heat shock protein genes.

The prevalence of canine babesiosis due to Babesia gibsoni has increased throughout the world including in southern India. The polymerase chain reaction (PCR) based molecular characterization of B. gibsoni in dogs of Kerala, south India, targeting three specific genes viz., apical membrane antigen (AMA1), 50 kDa surface antigen (P50), and heat shock protein (HSP70) was undertaken in this study. Out of 297 blood samples collected from clinically suspected animals, microscopy detected piroplasms of B. gibsoni in 60 (20.20 per cent), while the PCR targeting the BgP50 gene detected 85 (28.61 per cent). Polymerase chain reaction targeting the BgAMA1 and BgHSP70 detected a lesser number of samples (60 and 65 respectively) as positive. The phylogenetic analysis of BgHSP70 gene sequences did not reveal genetic heterogeneity among the B. gibsoni isolates of South India and from other countries, while the BgP50 gene differentiated the Indian isolates from Japanese isolates. When BgAMA1 was used for phylogenetic analysis, genetic variation was not observed among Indian and Taiwanese isolates, however, differentiated them from the Japanese isolates.

Morphological and molecular characterization of Rhipicephalus microplus and Rhipicephalus annulatus from selected states of southern India.

Ticks of the genus Rhipicephalus infesting cattle are the primary animal pests responsible for the annual economic loss of billions of dollars. Due to the morphological resemblance among the members of the Rhipicephalus (Boophilus) genus, species identification is very difficult. In this study, the adult R. annulatus and R. microplus ticks from two south Indian states viz., Kerala and Karnataka were subjected to morphological and molecular characterization. The R. microplus isolates from south India differed morphologically from true R. microplus clade A ticks. The ventral spur on the first pedipalp observed in male R. microplus was similar to that of R. australis. The phylogenetic analysis revealed that the R. microplus from these states clustered with R. microplus clade C. The mitochondrial cytochrome c oxidase I (COI) was identified as the preferred molecular marker compared to the internal transcribed spacer 2 (ITS2). The interspecific divergence between R. microplus and R. annulatus isolates from South India was 7.9 per cent based on COI. Moreover, based on COI, the R. microplus isolates revealed higher intraspecific divergence (2.9%) than R. annulatus (1%). The ITS2 sequences failed to differentiate R. microplus and R. annulatus.

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.

Sequence and phylogenetic analysis of the thrombospondin-related adhesive protein gene of Babesia gibsoni isolates in dogs in South India.

Babesia gibsoni, the causative agent of canine piroplasmosis, is a tick-borne intraerythrocytic protozoan parasite predominantly reported in Asian countries. The present study aimed at genotypic characterization of B. gibsoni isolates prevalent in dogs in Kerala, a southern state of India. Blood samples were collected from 272 dogs in Kerala and B. gibsoni infection was detected by microscopy and polymerase chain reaction (PCR). Molecular confirmation of B. gibsoni parasites was carried out by 18S rRNA nested-PCR, followed by sequencing. Nested-PCR detected a higher percentage of dogs (40.44%) positive for B. gibsoni infection than microscopy where 15.81% dogs were detected positive for infection. Genetic characterization of B. gibsoni isolates (n = 11) prevalent in dogs in the state of Kerala was carried out by PCR amplification and sequencing of the 855 bp thrombospondin-related adhesive protein (TRAP) gene fragment. Phylogenetic analysis of the B. gibsoni TRAP (BgTRAP) gene revealed that B. gibsoni isolates from Kerala formed a distinct cluster with the isolates from north India and Bangladesh, away from other East Asian isolates. Nucleotide analysis of the tandem repeats of BgTRAP gene showed considerable genetic variation among Indian isolates that was shared by B. gibsoni isolates of Bangladesh but not by the isolates of East Asian countries. The results of the present study further confirmed that B. gibsoni parasites in a distinct genetic clade are endemic in dogs in India and Bangladesh. However, elaborate studies are required for better understanding of the genetic diversity of B. gibsoni.

Prevalence of Anti-Toxocara canis Antibodies in Dogs Detected with Recombinant Cathepsin L-1 and TES-26 Antigens in Three States of India.

PURPOSE: Toxocara canis is a common intestinal nematode parasite of dogs with recognized zoonotic potential in tropical countries. The purpose of this study was to determine the seroprevalence of anti-T. canis antibodies in two target dog populations: household and community-owned, distributed over three distinct geographical regions of India. METHODS: Two recombinant proteins of T. canis, cathepsin L-1 (CL-1) and Toxocara excretory-secretory-26 (TES-26), expressed in Escherichia coli, were used for studying the prevalence of anti-T. canis antibodies in dog populations in three distinct geographical regions of the country using an IgG-enzyme-linked immunosorbent assay. A total of 615 sera, 507 from household and 108 from community owned dogs were screened for IgG antibodies. RESULTS: ELISA with recombinant (r) CL-1 showed 37.7% and 53.7% seroreactivity in household and community owned dogs, respectively. However, the rTES-26 antigen showed higher seroreactivity of 39.6% and 87.9% in the corresponding groups of household and community owned dogs, respectively. Chi-squared analysis of the data indicated that there was not any association in the prevalence of anti-T. canis antibodies between the samples analyzed from the three regions and the two cohorts of dog groups. However, the seroprevalence was higher in community owned dogs compared to household owned dogs. CONCLUSION: The results of the serological evaluation suggest that both the groups of dogs show high seroreactivity rates and are likely to harbor T. canis infections of tissue dwelling dormant larvae.

Reverse zoonosis of coronavirus disease-19: Present status and the control by one health approach.

The recent coronavirus disease (COVID-19) outbreak is one of its kind in the history of public health that has created a major global threat. The causative agent, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has a zoonotic source and hence, reverse zoonosis (disease transmission from humans to animals) increases the risk and rate of SARS-CoV-2 infection. Serological and molecular analyses and experimental infection studies have identified SARS-CoV-2 infection in several animal species in various countries. Different domestic and wild animals, including cats, dogs, tigers, lions, puma, snow leopard, minks, and pet ferrets, are infected naturally with SARS-CoV-2, mostly through suspected human to animal transmission. In addition, in vivo experimental inoculation studies have reported the susceptibility of cats, ferrets, hamsters, Egyptian fruit bats, and non-human primates to the virus. These experimentally infected species are found to be capable of virus transmission to co-housed animals of the same species. However, SARS-CoV-2 showed poor replication in livestock species such as pigs, chickens, and ducks with no detection of viral RNA after the animals were deliberately inoculated with the virus or exposed to the infected animals. As the pets/companion animals are more susceptible to COVID-19, the infection in animals needs an in-depth and careful study to avoid any future transmissions. The one health approach is the best inter-disciplinary method to understand the consequences of viral spread and prevention in novel host populations for the betterment of public health. Further in this review, we will explain in detail the different natural and experimentally induced cases of human to animal SARS-CoV-2 infection.

Comparative Expression Profiling and Sequence Characterization of ATP1A1 Gene Associated with Heat Tolerance in Tropically Adapted Cattle.

Climate change is an imminent threat to livestock production. One adaptation strategy is selection for heat tolerance. While it is established that the ATP1A1 gene and its product play an important role in the response to many stressors, there has been no attempt to characterize the sequence or to perform expression profiling of the gene in production animals. We undertook a field experiment to compare the expression profiles of ATP1A1 in heat-tolerant Vechur and Kasaragod cattle (Bos taurus indicus) with the profile of a heat-susceptible crossbreed (B. t. taurus × B. t. indicus). The cattle were exposed to heat stress while on pasture in the hot summer season. The environmental stress was quantified using the temperature humidity index (THI), while the heat tolerance of each breed was assessed using a heat tolerance coefficient (HTC). The ATP1A1 mRNA of Vechur cattle was amplified from cDNA and sequenced. The HTC varied significantly between the breeds and with time-of-day (p < 0.01). The breed-time-of-day interaction was also significant (p < 0.01). The relative expression of ATP1A1 differed between heat-tolerant and heat-susceptible breeds (p = 0.02). The expression of ATP1A1 at 08:00, 10:00 and 12:00, and the breed-time-of-day interaction, were not significant. The nucleotide sequence of Vechur ATP1A1 showed 99% homology with the B. t. taurus sequence. The protein sequence showed 98% homology with B. t. taurus cattle and with B. grunniens (yak) and 97.7% homology with Ovis aries (sheep). A molecular clock analysis revealed evidence of divergent adaptive evolution of the ATP1A1 gene favoring climate resilience in Vechur cattle. These findings further our knowledge of the relationship between the ATP1A1 gene and heat tolerance in phenotypically incongruent animals. We propose that ATP1A1 could be used in marker assisted selection (MAS) for heat tolerance.