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In the present study, we used a serum-free culture media to propagate goat putative spermatogonial stem cells (SSCs) and evaluated the effect of crucial growth factors on relative expression of some SSC markers and self-renewal related genes. The enriched SSCs were cultured on a homologous Sertoli cell feeder layer in KO-DMEM supplemented with 10% KOSR. Putative SSC colonies emerged between day 6 and 10 which were then characterized by the expression of numerous spermatogonial and pluripotency related markers. After 15 days of subculture, the relative mRNA expression study revealed that 40 ng/mL concentration of Glial cell line-derived neurotrophic factor (GDNF) upregulated the expression of BCL6B , ID4 , PLZF , and UCHL1 . Moreover, the supplementation of GDNF + bFGF up-regulated the expression of PLZF and BCL6B . UCHL1 expression was higher after addition of GDNF + LIF while, THY1 overexpressed in response to the addition of GDNF + CSF1. These results demonstrated that the goat SSCs were efficiently propagated using a KOSR based serum-free media and the growth factor supplementation markedly influences their gene expression profile.

Mobile genetic elements (MGEs) are associated with the emergence of multidrug resistance in extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae. This study explores the role of class 1 integrons and IS26 elements in breaching taxonomic barriers. A total of 110 E. coli bacteria were isolated from 300 clinical mastitis milk samples. The 98% E. coli isolates were extended-spectrum beta-lactamase- producers. About 83% of these isolates carried co-resistance for fluoroquinolones. The co-existence of (extended-spectrum beta-lactamase + quinolone resistance determining region mutations) and (extended-spectrum beta-lactamase + plasmid-mediated quinolone resistance genes) was found in 76% and 44% of isolates, respectively. The MGEs were detected in 88% of isolates with IS26 in 82% and class 1 integrase in 40% of isolates. The types of class 1 integron gene cassettes detected includes dfrA7, (dfrA17 + aadA5), and (dfrA1 + aadA1). We discovered 2 and 4 novel variants of the dfrA17 and aadA5 genes, respectively. We report a variant of aadA5 with mutation E235G in the Indian subcontinent earlier reported only in a human clinical isolate from Belgium. About 19 isolates carried IS26 linked to integrase gene intI1 with an internal deletion of 265 bp in the 5`CS of integrase gene intI1, earlier reported only in E. coli ST131 isolates from human clinical, wastewater samples. This study suggests intercontinental dissemination of antibiotic resistant genes (ARGs) across different microbiomes via mobile genetic elements.Key points• The role of mobile genetic elements in the emergence of multidrug-resistant E. coli in bovine mastitis.• Novel variants of the aadA5 (aminoglycoside adenyl transferase) and dfrA17 (dihydrofolate reductase) genes were identified in pathogenic E. coli isolated from bovine mastitis in class 1 integron gene cassette.• Sequence analysis of mobile genetic components revealed the physical connection between IS26 and intI1 genes with an internal deletion in 5'CS of class 1 integrase.

Aquaporins (AQPs) are integral membrane proteins responsible for water transport across cellular membranes in both prokaryotes and eukaryotes. A subfamily of AQPs, known as aquaglyceroporins (AQGPs), facilitate the transport of small solutes such as glycerol, water, and other solutes across cellular membranes. These proteins are involved in a variety of physiological processes, such as organogenesis, wound healing, and hydration. Although AQPs have been studied extensively in different species, their conservation patterns, phylogenetic relationships, and evolution in mammals remain unexplored. In the present study, 119 AQGP coding sequences from 31 mammalian species were analysed to identify conserved residues, gene organisation, and most importantly, the nature of AQGP gene selection. Repertoire analysis revealed the absence of AQP7, 9, and 10 genes in certain species of Primates, Rodentia, and Diprotodontia, although not all three genes were absent in a single species. Two Asparagine-Proline-Alanine (NPA) motifs located at the N- and C-terminal ends, aspartic acid (D) residues, and the ar/R region were conserved in AQP3, 9, and 10. Six exons encoding the functional MIP domain of AQGP genes were found to be conserved across mammalian species. Evolutionary analysis indicated signatures of positive selection in AQP7, 9, and 10 amongst different mammalian lineages. Furthermore, substitutions of certain amino acids located close to critical residues may alter AQGP functionality, which is crucial for substrate selectivity, pore formation, and transport efficiency required for the maintenance of homeostasis in different mammalian species.

Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen that poses a significant threat in cases of chronic mastitis in dairy animals. The ability of MRSA to persist in the host is attributed to various virulence factors, genes encoding surface adhesins, and determinants of antibiotic resistance, which provide it a survival advantage. This investigation focused to determine the virulence factors, antimicrobial resistance (AMR) profile and biofilm production potential of 46 MRSA isolates from 300 bovine mastitis milk samples. The AMR profile revealed a high level of resistance, with 46 and 42 isolates resistant to cefoxitin and oxacillin, respectively, followed by 24 and 12 isolates resistant to lomefloxacin and erythromycin, respectively. Only 2 isolates resistant to tetracycline and none were resistant to chloramphenicol. The study also evaluated various virulence factors such as coa (n = 46), nuc (n = 35) hlg (n = 36), pvl (n = 14), tsst-1(n = 28) spa (n = 39) and enterotoxin genes sea (n = 12) and seg (n = 28) and identified antibiotic resistance determinants mecA and blaZ in 46 and 27 isolates, respectively. Intercellular adhesion genes icaA and icaD were present in 40 and 43 isolates, respectively and surface adhesion genes ebps, fnbpA, eno, sasG, cna, and bap were found in 43, 40, 38, 26, 21 and 1 isolates, respectively. Microtiter plate (MTP) assay revealed that 29 MRSA isolates were capable of producing biofilms, whereas 17 were not. Biofilms producing MRSA isolates possessed adhesion genes, virulence factors, toxin genes and AMR genes that may act synergistically towards a chronic disease progression, illness and severe damage to the udder, which generally last for several months and very challenging to cure.

In order to ensure food safety, screening food samples for the presence of pathogens has been categorised as a legal testing item throughout the globe. One of the most prevalent zoonotic bacteria transmitted through dairy milk is Staphylococcus aureus. Given the limitations of the conventional detection methods, in the current study we desigined a competitive lateral flow immune assay (LFIA) using colloidal silver nanoparticles derived from mango leaves for the detection of Staphylococcus aureus in cow milk. SpA, a recombinant protein of Staphylococcus aureus, was used to raised hyperimmune sera used for developing the assay followed by conjugation with the synthesized nanoparticles. To increase the specificity of the assay, the milk samples were prenriched with selective agar exclusively require for Staphyloccocus aureus. The assay was found to be completed within 7–8 h by observing test and control lines in LFIA strips. The developed assay was found to specifically detect the bacteria as low as 1000 cfu/ml of milk samples. With a total 230 number of raw and clinical mastitis milk samples, the assay was validated and achieved relative accuracy, specificity, and sensitivity values of 97.39, 98.03, and 96.1%, respectively. The developed LFIA, which uses economically feasible and stable silver nanoparticles derived from mango leaves, has the potential for routine screening of milk samples for the presence of Staphylococcus aureus, especially in low-resource settings, allowing for early diagnosis, which facilitates effective treatment for the dairy animals and prevents the transmission of the disease in consumers.

Indicine cattle (Bos indicus) are known for resilience to infectious diseases and environmental stress. However, the genomic basis underlying this advantage remains poorly understood. To characterize variations in immune-related genetic elements in four indicine breeds (Kangayam, Gir, Tharparkar, and Sahiwal), a taurine breed (Holstein Friesian), and a taurine-indicine crossbreed (Karan Fries), we performed integrative whole-genome analysis. Using whole-genome data representing 108 animals, we identified structural variants, copy number variants, single-nucleotide variations, and insertions/deletions. High-impact single-nucleotide variations in the key innate immune genes, CARD9 and NLRP8, shared across all indicine breeds, were absent in the other breeds. Genetic differentiation analysis identified several innate immune genes showing strong divergence between the indicine breeds and the taurine breed. Selective sweep detection analysis highlighted multiple breed-specific immune-related sweep regions. Functional enrichment analysis showed significant enrichment of immune pathways in indicine breeds. A comparison of the candidate genes with basal gene expression profiles of unchallenged peripheral blood mononuclear cells indicated that genomic variation influences the differential expression of several genes in indicine breeds. We synthesized the data from population genome structure analysis, nucleotide diversity, genetic differentiation, selective sweep analysis, and correlations with gene expression profiles. Indicine breeds exhibited a higher number of immune-related variants and stronger signals of selection in immune pathways. These findings provide a curated set of innate immune gene candidates in indicine breeds for future functional studies and breeding programs.Competing Interest StatementThe authors have declared no competing interest.Footnotes* https://www.ebi.ac.uk/eva/?eva-study=PRJEB88461Funder Information DeclaredIndian Council of Agricultural Research, https://ror.org/04fw54a43, F. No.NASF/SUTRA-02/2022-23/50)

In this study, we focused on the rapid land use and land cover (LULC) changes in Bankura in 1990, 2000, 2010, and 2024, employing an integrated remote sensing, geospatial, and statistical approach to track land use changes. The supervised classification technique and change detection analysis were applied with the Supper Vector Machine (SVM), Maximum Likelihood (ML), and Random Forest (RF) methods to identify land use classes in various categories like Dense Forest, Open Forest, water body, agricultural land, settlement, barren land, and sand. The Kappa Coefficient was used for the accuracy assessment, which revealed that the overall accuracy of 1990 was 93.33%, 2000 was 93.23%, 2010 was 93.43%, and 2024 was 90%. The analysis revealed a significant increase in built-up land from agricultural and forested areas, with a higher percentage of agricultural land converted to built-up areas observed between 1990 and 2024. During this interval, the built-up land area increased by approximately 13.6%, primarily due to the conversion of agricultural land and forest cover. Agricultural land decreased by 11.45%, while dense forest cover declined by 7.75%, indicating a significant anthropogenic influence on landscape transformation. Our findings underscore the importance of sustainable land use planning, conservation efforts, and policy interventions in mitigating environmental degradation, leveraging the effectiveness of space-based inputs and geospatial techniques. The research emphasizes the need for continuous monitoring and further investigation into socio-economic drivers and environmental consequences to ensure resilient urban management and sustainable development. This reveals the importance of reforestation, preserving water bodies, and developing ecologically sensitive infrastructure. Moreover, the study highlights the importance of sustainable land use planning in mitigating adverse environmental impacts and preserving ecological balance.