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Abstract

Wild ruminants are an interesting topic for research because only limited information exists regarding their microbiota. They could also be an environmental reservoir of undesirable bacteria for other animals or humans. In this study faeces of the 21 free-living animals was sampled (9 Cervus elaphus-red deer, adult females, 12 Capreolus capreolus-roe deer, young females). They were culled by selective-reductive shooting during the winter season of 2014/2015 in the Strzałowo Forest District-Piska Primeval Forest (53° 36 min 43.56 sec N, 21° 30 min 58.68 sec E) in Poland. Buttiauxella sp. is a psychrotolerant, facultatively anaerobic, Gram-negative rod anaerobic bacte- rial species belonging to the Phylum Proteobacteria, Class Gammaproteobacteria, Order Entero- bacteriales, Family Enterobacteriacae and to Genus Buttiauxella. Buttiauxella sp. has never previ- ously been reported in wild ruminants. In this study, identification, antimicrobial profile and sensitivity to enterocins of Buttiauxella strains were studied as a contribution to the microbiota of wild animals, but also to extend knowledge regarding the antimicrobial spectrum of enterocins. Five strains were identified using the MALDI-TOF identification system (evaluation score value was up to 2.224) and allotted to the genus Buttiauxella including the species Buttiauxella gaviniae, B. ferragutiae, B. agrestis. Strains were DNase negative, and they hydrolysed esculin; fermentation of L-arabinose, D-mannitol and D-mannose was positive. Dulcitol, inositol reaction, urea and indol were negative. Buttiauxella strains did not form biofilm. They were resistant to at least one of the 13 antibiotics tested. B. agrestis 2/109/1 was resistant to amdinocillin, clindamycin and pen- icillin. However, Buttiauxella strains were sensitive to the enterocins used (inhibition activity ranged from 100 to 25 600 AU/ml).
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Abstract

Enterococcus hirae belongs in the Enterococcus faecium group within the genus Enterococcus. This species occurs naturally in the environment, commensally in the alimentary tracts of animals, and pathologically for example in humans with urinary infections. Some strains of E. hirae possess virulence factors, including biofilm formation. Biofilm growth protects bacteria against host de- fences; biofilm can be a source of persistent infection. Testing bacterial strains for their ability to form biofilm might therefore facilitate their treatment or prevention. This study focuses on bio- film formation by E. hirae strains derived from various animals. This kind of testing has never been done before. A total of 64 identified E. hirae from laying hens, ducks, pheasants, ostriches, rabbits, horses and a goat were tested by means of three methods; using Congo red agar, the tube method and microtiter plate agar. The majority of strains were found to form biofilm. 62.5% of strains were biofilm-forming, four categorized as highly positive (OD570 ≥1); most strains were low-grade biofilm positive (0.1 ≤ OD 570 < 1). Related to poultry, 55 E. hirae strains were tested nd found to produce biofilm; 24 strains did not form biofilm, 31 strains were biofilm-forming; 27 strains showed low-grade biofilm formation, and four strains were highly biofilm-forming. Four strains from hens and ostriches reached the highest OD570 values, more than 0.500. Rabbit-derived E. hirae strains as well as strains isolated from horses and the goat were low-grade bio- film-forming. Microtiter plate assay proved to be the best tool for testing the in vitro biofilm for- mation capacity of E. hirae strains from different species of animals.
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