Phylogeny, Multiple Antibiotic Resistance Index and BiofilmCharacterization of Mannheimia species Isolated from ClinicalCases of Pneumonia in Small Ruminants
DOI:
https://doi.org/10.48165/ijvsbt.20.2.02Keywords:
Biofilms, Mannheimia, MAR-index, 16S rRNA, PhylogenyAbstract
Abstract
Mannheimia is a Gram-negative, bipolar organism that is frequently involved in respiratory infections of bovines and small ruminants. In the present study, a total of 92 nasal swab samples were examined from enzootic pneumonia cases of sheep and goat, out of which 30 Mannheimia isolates have been identified. The 16S rRNA sequence analysis of the isolate revealed a close genetic distance between the Mannheimia species from different geographical regions. However, it formed a separate clade in the phylogenetic tree and the isolate is closely related to the M. hemolytica and M. caviae species. Three methods (Standard Tube method, Congo Red method and Microtiter Plate assay) were evaluated for identifying the biofilm forming Mannheimia isolates. Among them, Microtiter plate assay was the best method for quantification of biofilm production, by which 29 out of 30 isolates were identified as biofilm producers. In antibiogram study, the Mannheimia isolates exhibited resistance towards ampicillins, gentamicin, co-trimoxazole, tetracyclines and amoxicillin. All 30 isolates developed a multiple antibiotic resistance index (MAR) of more than 0.2. This indicates that there is a potential threat of antimicrobial resistance (AMR) transmission through food animals. In conclusion our study provides insights into the genetic relation between Mannheimia from India with other Mannheimia species from a different geographical region, and their capabilities of biofilm production.
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Abate,F.M. and Fentie Kassa,T. (2023). Isolation and identification of Mannheimia haemolytica and Pasteurella multocida from symptomatic and asymptomatic sheep and their antibiotic susceptibility patterns in three selected districts of north Gondar zone, Gondar Ethiopia. Veterinary Medicine and Science.
Bkiri,D., Semmate,N., Boumart,Z., Safini,N., Fakri,F.Z., Bamouh,Z. and El Harrak,M.(2021). Biological and molecular characterization of a sheep pathogen isolate of Mannheimia haemolytica and leukotoxin production kinetics. Veterinary World, 14(8): 2031.
Boukahil, I. and Czuprynski, C. J. (2016). Mannheimia haemolytica biofilm formation on bovine respiratory epithelial cells. Veterinary microbiology, 197, 129-136.
Boudewijns,M., Bakkers,J.M., Sturm,P.D. and Melchers,W.J.(2006). 16S rRNA gene sequencing and the routine clinical microbiology laboratory: a perfect marriage? J Clin Microbiol.Sep;44(9):3469-70.
Bauer,A.W (1996). Antibiotic susceptibility testing by a standardized single disc method. Am. J. of Clinc. Path., 45, 149-158.
Catry,B., Decostere,A., Schwarz,S., Kehrenberg,C., de Kruif,A. and Haesebrouck,F. (2006). Detection of tetracycline-resistant and susceptible Pasteurellaceae in the nasopharynx of loose group-housed calves. Veterinary research communications, 30, 707-715.
Christensen,H. and Bisgaard,M.(2010). Molecular classification and its impact on diagnostics and understanding the phylogeny and epidemiology of selected members of Pasteurellaceae of veterinary importance. Berliner Und Munchener Tierarztliche Wochenschrift, 123(1-2): 20-30.
Dhanawade,N.B., Kalorey,D.R., Srinivasan,R., Barbuddhe,S.B. and Kurkure,N.V.(2010). Detection of intercellular adhesion genes and biofilm production in Staphylococcus aureus isolated from bovine subclinical mastitis. Veterinary research communications, 34, 81-89.
Depenbrock,S., Aly,S., Wenz,J., Williams,D., ElAshmawy,W., Clothier,K. and Chigerwe,M. (2021). In-vitro antibiotic resistance phenotypes of respiratory and enteric bacterial isolates from weaned dairy heifers in California. PloS one, 16(11): e0260292.
Gharibi,D., Ghorbanpoor,M., Jabbari,A.R. and Cid,D. (2021). Molecular characterization of Mannheimia haemolytica associated with ovine and caprine pneumonic lung lesions. Microbial Pathogenesis, 153, 104791.
Marru,H.D., Anijajo,T.T. and Hassen,A.A.(2013). A study on Ovine pneumonic pasteurellosis: Isolation and Identification of Pasteurellae and their antibiogram susceptibility pattern in Haramaya District, Eastern Hararghe, Ethiopia. BMC Veterinary Research, 9(1): 1-8.
Önat,K., Kahya,S. and Carli,K.T.(2010). Frequency and antibiotic susceptibility of Pasteurella multocida and Mannheimia haemolytica isolates from nasal cavities of cattle. Turkish Journal of Veterinary & Animal Sciences, 34(1): 91-94.
Ocak,F. and Turkyilmaz,S.(2022). Investigation of Antimicrobial Resistance, Biofilm Production, Biofilm Associated Virulence Genes and Integron Genes of Pseudomonas aeroginosa Isolates Obtained from Animal Clinical Samples. Israel Journal of Veterinary Medicine, 77(1): 15-26.
Olson,M.E., Ceri,H., Morck,D.W., Buret,A.G. and Read,R.R.(2002). Biofilm bacteria: formation and comparative susceptibility to antibiotics. Canadian journal of veterinary research, 66(2): 86.
Ponnusamy,P., Masilamoni,B.S., Ranjith,K.M. and Manickam,R. (2017). Isolation, identification and antibiogram of Mannheimia haemolytica associated with caprine pneumonia in the Cauvery Delta Region of Tamil Nadu. Int J Curr Microbiol Appl Sci, 6(9): 3118-3122.
Post,K.W., Cole,N.A. and Raleigh,R.H.(1991). In vitro antimicrobial susceptibility of Pasteurella haemolytica and Pasteurella multocida recovered from cattle with bovine respiratory disease complex. Journal of Veterinary Diagnostic Investigation, 3(2): 124-126.
Pillai,D.K., Cha,E. and Mosier,D. (2018). Role of the stress-associated chemicals norepinephrine, epinephrine and substance-P in dispersal of Mannheimia haemolytica from biofilms. Veterinary microbiology, 215, 11-17.
Sahay,S., Natesan,K., Prajapati,A., Kalleshmurthy,T., Shome,B.R., Rahman,H. and Shome,R. (2020). Prevalence and antibiotic susceptibility of Mannheimia haemolytica and Pasteurella multocida isolated from ovine respiratory infection: A study from Karnataka, Southern India. Veterinary World, 13(9): 1947.
Singh,F., Sonawane,G.G. and Meena,R.K. (2019). Pathology, isolation and characterisation of virulent and diverse Mannheimia haemolytica and Pasteurella multocida associated with fatal pneumonia in sheep, Rajasthan, India. Comparative Clinical Pathology, 28, 531-540.
Seker,E., Kuyucuoglu,Y. and Konak,S. (2009). Bacterial examinations in the nasal cavity of apparently healthy and unhealthy Holstein cattle. Journal of Animal and Veterinary Advances, 8(11): 2355-2359.
Tamura,K., Stecher,G. and Kumar,S. (2021). MEGA11: molecular evolutionary genetics analysis version 11. Molecular biology and evolution, 38(7): 3022-3027.
Tremblay,Y.D., Deslandes,V. and Jacques,M. (2013). Actinobacillus pleuropneumoniaegenes expression in biofilms cultured under static conditions and in a drip-flow apparatus. BMC genomics, 14(1),
Tang,X., Zhao,Z., Hu,J., Wu,B., Cai,X., He,Q., & Chen,H. (2009). Isolation, antimicrobial resistance, and virulence genes of Pasteurella multocida strains from swine in China. Journal of clinical microbiology, 47(4): 951-958.
Van Nguyen,P., Le,C.T., Ho, X.T.T., Truong,P.H., Van Loi,B. and Nguyen, K.C.T. (2023). First Report of Antimicrobial Resistance of Mannheimia haemolytica from Phan Rang Sheep in Vietnam. Pakistan Veterinary Journal, 43(1).
Yılmaz,E.S. and Güvensen,N.C. (2016). In vitro biofilm formation in ESBL producing Escherichia coli isolates from cage birds. Asian Pacific journal of tropical medicine, 9(11): 1069-1074.
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