Phenotypical characterisation of intestinal spirochaetes isolated from pigs

Fellström, Claes; Gunnarsson, A.

doi:10.1016/0034-5288(95)90021-7

Cited by 118 publications

(112 citation statements)

References 19 publications

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“…Following selective culture, the isolates had been subcultured once, and if necessary subcultured to pure genotypes by 10-fold serial dilution as described elsewhere (Jansson et al, 2008). The isolates were stored in liquid nitrogen (Fellströ m & Gunnarsson, 1995) in the strain collection of the National Veterinary Institute (SVA), Uppsala, Sweden until further use.…”

Section: Methodsmentioning

confidence: 99%

Antimicrobial susceptibility ofBrachyspiraspp. isolated from commercial laying hens and free-living wild mallards (Anas platyrhynchos)

Jansson¹,

Pringle

2011

Avian Pathology

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In vitro antimicrobial susceptibility to tylosin, valnemulin, tiamulin, doxycycline, lincomycin and ampicillin was investigated by broth dilution in 48 Brachyspira spp. isolates from commercial laying hens (n030) and free-living wild mallards (Anas platyrhynchos) (n018). Presumed pathogens (Brachyspira alvinipulli, Brachyspira intermedia, Brachyspira pilosicoli), commensals (Brachyspira murdochii, Brachyspira innocens, ''Brachyspira pulli''), and isolates of undetermined species affiliation were included. The laying hens had not been exposed to therapeutic levels of antimicrobials for at least 50 weeks before sampling, and low levels of environmental antimicrobial exposure were presumed in mallards. No isolates with decreased susceptibility to tylosin, valnemulin, tiamulin or doxycycline were found. Decreased susceptibility to lincomycin (minimum inhibitory concentration 16 mg/ml) was detected in two isolates (Brachyspira sp.) from laying hens. Five isolates showed decreased susceptibility to ampicillin (minimum inhibitory concentration 16 to !32 mg/ml), including two ''B. pulli'' and one B. alvinipulli from laying hens, and isolates of B. pilosicoli and ''B. pulli'' from mallards. Decreased susceptibility to ampicillin was associated with b-lactamase activity in four isolates. A new variant of a class D b-lactamase gene designated bla oxa-192 was identified in a B. pilosicoli isolate of mallard origin. This is the first time the genetic basis for antimicrobial resistance is described in Brachyspira spp. from a free-living wild bird. Isolates displaying decreased susceptibility to ampicillin were accompanied by fully susceptible isolates of the same species or other genotypes within three laying hen flocks. This underlines the need for performing antimicrobial susceptibility tests on single clones/genotypes, and to analyse multiple isolates from the same flock.

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Section: Methodsmentioning

confidence: 99%

Antimicrobial susceptibility ofBrachyspiraspp. isolated from commercial laying hens and free-living wild mallards (Anas platyrhynchos)

Jansson¹,

Pringle

2011

Avian Pathology

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“…Three isolates of B. pilosicoli were biochemically atypical (galactosidase negative and β-glucosidase positive. ), 6 isolates of B. pilosicoli were biochemically identified and consequently placed in the group IVa, characteristic for B. pilosicoli (Fellström et al 1995). All the isolates of B. pilosicoli hydrolyzed sodium hippurate (Table 1).…”

Section: Resultsmentioning

confidence: 99%

“…The type of haemolysis was always specified after 4 days of cultivation on the Tryptose Soy Agar with blood. Bacterial cultures were biochemically identified on the basis of sodium hippurate hydrolysis determination and hydrolytic activity of enzymes (α-glucosidase, β-glucosidase and α-galactosidase) with the use of the diagnostic tablets (A/S Rosco, Denmark) (Fellström et al 1995). The control strains of B. hyodysenteriae B78 (ATCC 27164 T ), B. pilosicoli (ATCC 51139 T ) were used to control biochemical reactions.…”

Section: Methodsmentioning

confidence: 99%

Isolation and Characterization of Brachyspira spp. from Dogs in the Czech Republic

2010

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The objectives of this study were to establish the prevalence of intestinal Spirochetes of the genus Brachyspira in Czech dogs and to determine the susceptibility of obtained B. pilosicoli isolates to selected antibacterial substances. Spirochetes were diagnosed microscopically in 23 out of 1139 samples of dogs' excrements, primarily intended for a parasitological testing. The cultivation of positive samples provided 10 brachyspira isolates, which were, on the basis of their biochemical activity and the results of the species-specific PCR, identified as B. pilosicoli (9 isolates) and B. hyodysenteriae (1 isolate). These dogs came from households. All the 7 tested isolates B. pilosicoli were sensitive to metronidazole and doxycycline, uniformly resistant to erythromycin, partly sensitive to cefazoline, lincomicine and ampicilline except for one isolate of B. pilosicoli, which was resistant to ampicilline.The second part of study was focused on dogs with diarrhoea that came from animal shelters, where a high prevalence of 58% (10/17) of B. pilosicoli was found. Spirochetal colitis, susceptibility testingIn the past, spirochetes have been observed and isolated from colonic excrements and mucosa of both healthy dogs and dogs with diarrhoea. However, the pathogenic importance of spirochetes and their role in the onset of canine diarrhoea have not been sufficiently explained until now (Pindak et al. 1965; Turek and Meyer 1978; Meyer et al. 1982;Lee and Hampson 1994;Duhamel et al. 1995, 1997, Fellstrom et al. 2001Oxberry and Hampson 2003). Weakly haemolytic and indole-negative spirochetes colonising colon and rectum of healthy dogs were characterised by PCR method specific for 16S rRNA and MLEE (multilocus enzyme electrophoresis) and classified as a new species designated as "Brachyspira canis". These spirochetes probably represent common canine intestinal commensals (Duhamel et al. 1998;Oxberry and Hampson 2003). Another species that has been isolated from canine and human faeces and the faeces of various animals in connection with diarrhoea and colonic infections is Brachyspira pilosicoli (Trott et al. 1996). This species tends to be detected in puppies with diarrhoea affected by a parasitic coinfection of Giardia spp. In adult dogs, it represents frequent finding also in the individuals without clinical symptoms (Turek and Meyer 1978;Duhamel et. al 1996). The cultures isolated from dogs were able to colonise chicken enterocytes via a mechanism similar to the one described in isolates of B. pilosicoli obtained from humans and pigs (Muniappa et al. 1996), which implies their pathogenic potential. Brachyspira pilosicoli is nowadays considered to be the causative agent of canine intestinal spirochetosis. Data on Brachyspira isolation from dogs came from studies on smaller groups of dogs or from sporadic cases of enteric infections documented more in detail (Oxberry and Hampson 2003; Skrzypcak et al. 2007). Reports on susceptibility of B. pilosicoli originating from dogs have appeared only rarely until now (Adac...

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“…These 4 species of weakly beta-hemolytic intestinal spirochete have a similar appearance on blood agar, and the pathogenicity of these species except B. pilosicoli is controversial. Although Kinyon and Harris [8] and Lee et al [9] considered these species as non-pathogenic, other studies [2,10] have provided evidence that even these species may induce enterocolitis.…”

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confidence: 99%

Identification of Weakly Beta-Hemolytic Porcine Spirochetes by Biochemical Reactions, PCR-Based Restriction Fragment Length Polymorphism Analysis and Species-Specific PCR

Ohya

Araki²,

Sueyoshi

2008

The Journal of Veterinary Medical Science

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ABSTRACT. We examined the usefulness of PCR-based restriction fragment length polymorphism (PCR-RFLP) and species-specific PCR combined with a newly devised rapid biochemical test using microplates for identifying weakly beta-hemolytic intestinal spirochetes (WBHIS) isolated from pigs. WBHIS strains showing atypical biochemical characteristics were decisively identified at the species level by PCR-RFLP and species-specific PCR. Identification of WBHIS at the species level in routine diagnostic work will certainly contribute to clarifying the pathogenicity of WBHIS. KEY WORDS: Brachyspira, PCR-RFLP, spirochete.J. Vet. Med. Sci. 70(8): 837-840, 2008 Intestinal infections with Brachyspira species are responsible for severe economic losses in the swine industry worldwide. Brachyspira species identified in pigs include one strongly hemolytic species Brachyspira hyodysenteriae and 4 weakly beta-hemolytic intestinal spirochetes (WBHIS): B. innocens, B. pilosicoli, B. intermedia, and B. murdochii. B. hyodysenteriae is considered the most pathogenic species and causes swine dysentery [6,17]. B. pilosicoli has been linked to a milder colitis known as porcine intestinal (or colonic) spirochetosis [5,14,15]. These 4 species of weakly beta-hemolytic intestinal spirochete have a similar appearance on blood agar, and the pathogenicity of these species except B. pilosicoli is controversial. Although Kinyon and Harris [8] and Lee et al. [9] considered these species as non-pathogenic, other studies [2, 10] have provided evidence that even these species may induce enterocolitis.Our bacteriological surveys of swine suffering from swine dysentery, colitis, or diarrhea, indicated that 49 strains of WBHIS were isolated in 21 cases and B. hyodysenteriae was isolated in 101 cases. Among the 21 cases, B. hyodysenteriae was isolated with WBHIS in three cases, and B. pilosicoli and B. innocens were isolated in one case. In the remaining 17 cases, only a single species of WBHIS was isolated.Bacteriological examination of swine dysentery and related diseases is mainly focused on B. hyodysenteriae or B. pilosicoli. Moreover, precise identification of WBHIS tends to be neglected, and this may correlate to the insufficient description of their pathogenicity.The aim of this study was to evaluate the usefulness of PCR-based restriction fragment length polymorphism (PCR-RFLP) and species-specific PCR combined with biochemical tests for identifying WBHIS. Forty nine isolates of WBHIS were investigated in this study, and 4 additional type strains of Brachyspira species were included as references (Table 1).Currently, porcine Brachyspira isolates are differentiated based on phenotypic criteria, including intensity of hemolysis, indole production, hippurate hydrolysis, and the activities of α-galactosidase, α-glucosidase, and β-glucosidase [3,12].The spirochetes were streaked on trypticase soy agar (TSA; Becton Dickinson Microbiology Systems, Sparks, MD., U.S.A.) containing 5% sheep blood and incubated for three days under anaerobic conditions....

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Phenotypical characterisation of intestinal spirochaetes isolated from pigs

Cited by 118 publications

References 19 publications

Antimicrobial susceptibility ofBrachyspiraspp. isolated from commercial laying hens and free-living wild mallards (Anas platyrhynchos)

Antimicrobial susceptibility ofBrachyspiraspp. isolated from commercial laying hens and free-living wild mallards (Anas platyrhynchos)

Isolation and Characterization of Brachyspira spp. from Dogs in the Czech Republic

Identification of Weakly Beta-Hemolytic Porcine Spirochetes by Biochemical Reactions, PCR-Based Restriction Fragment Length Polymorphism Analysis and Species-Specific PCR

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