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. 2022 Dec 26;18(1):452.
doi: 10.1186/s12917-022-03546-4.

Genotypic and phenotypic diversity of Prototheca spp. recovered from bovine mastitis in terms of antimicrobial resistance and biofilm formation ability

Niloufar Tashakkori  1 , Hamideh Kalateh Rahmani  2 , Babak Khoramian  3
Affiliations

Genotypic and phenotypic diversity of Prototheca spp. recovered from bovine mastitis in terms of antimicrobial resistance and biofilm formation ability

Niloufar Tashakkori et al. BMC Vet Res. .

Abstract

Background: The Prototheca algae have recently emerged as an important cause of bovine mastitis globally. Isolates from bovine mastitis in several countries were nearly all identified as P. bovis, suggesting that it was the main causative agent of bovine protothecal mastitis. The aim of the present study was to evaluate the presence and isolation of Prototheca spp. in dairy farms, detect the genetic diversity among strains, determine the capacity of producing biofilm and their resistance to antifungal and antimicrobial drugs.

Results: A total of 48 Prototheca isolates from four different farms were randomly selected to be investigated. Multiplex PCR showed all isolated colonies were Prototheca bovis. Performing RAPD-PCR by using OPA-4 primer, it was revealed that there was a clear amplification pattern. Different levels of biofilm production were observed among strains. Among 48 isolates, only 4 of them (8.33%) showed strong biofilm production. By using E-test strips, amphotericin B was able to inhibit the growth of all the strains tested. Disc diffusion method used for antimicrobial sensitivity test showed that the highest activity was demonstrated by gentamicin and colistin with 95.83% (46/48) and 89.58% (43/48) of sensitive strains, respectively.

Conclusions: The present study showed that RAPD-PCR was a rapid tool for discriminating P. bovis strains. Also, gentamicin and colistin can be considered as potential antimicrobial drugs which can prevent the growth of the mentioned strains in vitro, although there is no effective clinical treatment yet. Further studies are needed in order to detect an effective clinical therapy considering biofilm production by Prototheca spp. and their probable role in Prototheca pathogenicity.

Keywords: Antifungal; Antimicrobial; Biofilm; Molecular typing; Prototheca; RAPD-PCR.

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Conflict of interest statement

The authors declare that they have no conflict of interest.

Figures

Fig. 1
Fig. 1
Strains divided by genetic differences shown by dendrogram, farm and season of isolation, MIC, antibiotic susceptibility, biofilm formation capacity. W: weak biofilm producer. M: medium biofilm producer. S: strong biofilm producer. N: negative. MIC: Minimum Inhibitory Concentration. NFX: Enrofloxacin, SLT: Sulfonamide + Trimethoprim, CRO: ceftriaxone, GM: gentamicin, S: streptomycin, T: oxytetracycline, L: lincomycin, TY: tylosin, FF: florfenicol, AMX: amoxicillin, P: penicillin, CL: colistin, TE: tetracycline
Fig. 2
Fig. 2
MIC using Amphotericin B strip and symmetrical inhibition ellipse centered along the strip
Fig. 3
Fig. 3
Antibiotic susceptibility test using disc diffusion method performed on Mueller Hinton agar
See this image and copyright information in PMC

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