Dataset Information

Identification of coagulase negative staphylococcal species from bovine mastitis in India.

ABSTRACT:

Background

Staphylococcal mastitis is a major cause of concern to the dairy industry in India and several countries worldwide. Though Staphylococcus aureus is the major cause, coagulase negative staphylococcal species (CoNS) are being increasingly reported in recent years.

Aims

To investigate the incidence of coagulase negative staphylococcal species in bovine mastitis.

Methods

Isolation of staphylococci was carried out from 237 milk samples of cows and She buffaloes with clinical and subclinical mastitis from different regions of Andhra Pradesh and Karnataka. CoNS isolates were identified by tube coagulase test using fresh rabbit plasma and coagulase gene PCR. We employed the biochemical test scheme published elsewhere previously for identification of the CoNS isolates up to species and subspecies levels. Seven representative isolates were identified by 16S rDNA sequencing to check the accuracy of biochemical test based identification.

Results

The CoNS constitute the majority of the staphylococcal isolates from mastitis (80/125, 64%) in this region. Using biochemical test scheme, the CoNS isolates from bovine mastitis were identified as S. cohnii sub sp. cohnii, S. simulans, S. capitis sub sp. capitis, S. cohnii sub sp. xylosus, and S. lugdunensis. The CoNS species S. schleiferi, S. haemolyticus, S. sciuri, S. xylosus, S. chromogenes, and Macrococcus epidermidis were identified by 16S rDNA sequencing.

Conclusion

The 16S rDNA sequencing is the appropriate method for the identification of CoNS species. This study highlighted coagulase negative staphylococcal species as possible etiological agents of mastitis.

SUBMITTER: Bhavana RN

PROVIDER: S-EPMC9984147 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature

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Publications

Identification of coagulase negative staphylococcal species from bovine mastitis in India.

Bhavana R N RN Chaitanya R K RK

Iranian journal of veterinary research 20220101 4

<h4>Background</h4>Staphylococcal mastitis is a major cause of concern to the dairy industry in India and several countries worldwide. Though <i>Staphylococcus aureus</i> is the major cause, coagulase negative staphylococcal species (CoNS) are being increasingly reported in recent years.<h4>Aims</h4>To investigate the incidence of coagulase negative staphylococcal species in bovine mastitis.<h4>Methods</h4>Isolation of staphylococci was carried out from 237 milk samples of cows and She buffaloes ...[more]

PMID: 36874184

Similar Datasets

Project description:AimThis study was devoted to elucidate the tetracycline resistance of coagulase-negative staphylococci (CNS) derived from normal and subclinical mastitic (SCM) buffaloes' milk in Egypt.Materials and methodsA total of 81 milk samples from 46 normal buffalo milk samples and 35 SCM buffalo milk samples at private dairy farms of Egypt were used in this study. CNS were identified using phenotypic and molecular methods (polymerase chain reaction [PCR]). CNS isolates were tested for tetracycline resistance using routine methods and multiplex PCR targeting tetracycline (tet) resistance genes followed by sequencing of positive PCR products and phylogenetic analysis.ResultsIsolation and identification of 28 (34.5%) CNS from normal and SCM buffaloes' milk, namely, Staphylococcus intermedius (39.2%), Staphylococcus xylosus (25.0%), Staphylococcus epidermidis (10.7%), Staphylococcus hominis (10.7%), and 3.5% to each of Staphylococcus sciuri, Staphylococcus hyicus, Staphylococcus lugdunensis, and Staphylococcus simulans. Using nested PCR, all the 28 CNS isolates revealed positive for 16srRNA gene specific for genus staphylococci and negative for thermonuclease (nuc) gene specific for Staphylococcus aureus species. The presence of tetracycline resistance-encoding genes (tetK, tetL, tetM, and tetO) was detected by multiplex PCR. All isolates were negative for tetL, M, and O genes while 14 (50%) CNS isolates were positive for tetK gene, namely, S. lugdunensis (100%), S. hominis (100%), S. epidermidis (66.6%), S. intermedius (45.4%), and S. xylosus (42.8%). Nucleotide sequencing of tetK gene followed by phylogenetic analysis showed the high homology between our CNS isolates genes of tetracycline resistance with S. aureus isolates including Egyptian ones. This proves the transfer of the tetracycline resistance encoding genes between coagulase-negative and coagulase positive Staphylococcus spp.ConclusionCNS isolates have distinguishingly high resistance to tetracycline. Abundant tetracycline usage for mastitis treatment leads to the spread of genetic resistance mechanisms inside CNS strains and among all Staphylococcus spp. Consequently, tetracycline is not effective anymore.

Project description:Purpose: The aim of the study was to compare the miRNA expression in non-infected (H) mammary gland parenchyma samples with that of glands infected with coagulase-positive staphy lococci (CoPS) or coagulase-negative staphylococci (CoNS) using next-generation sequencing. Methods: miRNA-seq analysis was performed on mammary gland parenchyma samples collected from non-infected cows and those infected with coagulase-positive or -negative staphylococci. The miRNA libraries were constructed from total RNA using NEBNext Multiplex Small RNA Library Prep Set for Illumina (New England Biolabs) according to the manufacturer protocol. The quantification of the obtained libraries was performed on a Qubit 2.0 spectrophotometer (Invitrogen, Life Technologies), while a quality control on a TapeStation 2200 instrument (D1000 ScreenTape; Agilent). Single-end cycle sequencing was performed on the HiScanSQ platform (Illumina) with the use of TruSeq SR Cluster Kit v3- CBOT-HS and TruSeq SBS Kit v 3 - HS (Illumina). MicroRNA differentially expressed between investigated groups were identified with the DESeq2 software. Results: Comparing the CoPS and H groups, 256 known and 260 potentially new miRNAs were identifed, including 32 that were diferentially expressed (p≤0.05), of which 27 were upregulated and 5 downregulated. Comparing the CoNS and H groups, 242 known and 171 new unique miRNAs were identifed: 10 were upregulated (p≤0.05), and 2 downregulated (p≤0.05). Comparing CoPS with H and CoNS with H, 5 Kyoto Encyclopedia of Genes and Genomes pathways were identifed; in both comparisons, diferentially-expressed miRNAs were associated with the bacterial invasion of epithelial cells and focal adhesion pathways. Four gene ontology terms were identifed in each comparison, with 2 being common to both immune system processes and signal transduction. Conclusions: Obtained results enabled us to characterize the miRNA profile of the mammary gland parenchyma tissue, not only the healthy one but also the tissue infected with coagulase-positive and negative staphylococci as well as allowed identification of miRNAs differing the examined groups and characteristic for the staphylococci infection. They also indicated that miRNAs, especially miR-99 and miR-182, play an essential role in the epigenetic regulation of a range of cellular processes, including immunological systems bacterial growth in dendritic cells and disease pathogenesis (miR-99), DNA repair and tumor progression (miR-182).

Dataset Information

Identification of coagulase negative staphylococcal species from bovine mastitis in India.

Background

Aims

Methods

Results

Conclusion

Publications

Identification of coagulase negative staphylococcal species from bovine mastitis in India.

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