Asit Kumar Chakraborty*, Uttam Maity and Chandan Halder

Department of Biotechnology and Biochemistry, Oriental Institute of Science and Technology-West Bengal, Vidyasagar University, India

*Corresponding Author: Asit Kumar Chakraborty, Department of Biotechnology and Biochemistry, Oriental Institute of Science and Technology-West Bengal, Vidyasagar University, India.

Received: March 14, 2023; Published: September 29, 2023

Abstract

Vibrio species contamination in fish is a serious threat to human population worldwide. Aquaculture of shrimp has increased in India due to high demand in the Europe and America. Previously, we reported the presence of unique chromosomal blaCARB, pbp1B and CatC1 mdr genes involved in multi-resistance of Vibrio parahaemolyticus (Vp) and useful for diagnostic primers design. Escherichia coli AcrA-AcrB-TolC tripartite multidrug efflux genes were related to MexA-MexB-OprM genes of Pseudomonas aeruginosa and OqxAB-TolC genes of Klebsiella pneumoniae. Here, we showed that VmeAB and VmeYZ related to acrAB or mexAB as well as MacAB drug-efflux genes might be also responsible for multidrug resistance in Vp. Many heterogeneous Vme-isomers like vmeAB, vmeCD, vmeEF, vmeHI, vmeJK located in Ch-1 whereas vmeHI and vmeCD located in the complement strand (accession no. CP034294). The Ch-2 of Vp (accession no. CP020428) contained vmeYZ, vmePQ, vmeUV (RND-type) as well as macAB (ABC-type), EmrD (MFS-type) and mdtL (MATE-type) drug efflux genes. BLAST-2 analysis suggested only vmeB and vmeZ had similarities over 40% to acrB, mexB or oqxB types popular bacterial RND permease subunits. The macB protein of Vp has 56% similarity to E. coli macB protein and within the related Vibrio species (V. harveyi and V. alginolyticus) the similarity found to be round 98%. The Vp EmrD protein has 41.49% homology to E. coli while V. cholerae protein has only 29% homology. The MdtL protein of Vp has 61% similarity to E. coli and not suitable for primer design. We designed the primers using NCBI Primer Design Software and oligonucleotides were validated by Oligo Analyzer Software 3.2. Multi-alignment of vme-related sequences done by CLUSTAL-Omega software to confirm heterogeneities and BLASTN search of primers confirmed specificities to Vp Ch-1 or Ch-2. The P2F-vmeB/P2.1R-vmeB primers for VmeB gene and P2.1F-macB/P2R-macB primers for MacB gene should be useful with good species specificity. Such primers will be useful to detect V. parahaemolyticus contamination in fish aquaculture.

Keywords: Multidrug Efflux Genes; Diagnostic PCR Primers; Vibrio parahaemolyticus; Shrimp Aquaculture; Multidrug Resistance; acrAB and mexAB, macAB and emrCD

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Citation

Citation: Asit Kumar Chakraborty., et al. “Characterization of Chromosomal VmeAB, MacAB and EmrD Multidrug-Efflux Genes of Vibrio parahaemolyticus to Design Diagnostic PCR Primers to Improve Associated Problems in Shrimp Aquaculture”.Acta Scientific Medical Sciences 7.10 (2023): 71-85.

Copyright

Copyright: © 2023 Asit Kumar Chakraborty., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.