Differentiation of Serpulina species by NADH oxidase gene (nox) sequence comparisons and nox-based polymerase chain reaction tests

Atyeo, R.F.; Stanton, Thad B.; Jensen, Neil S.; Suriyaarachichi, D.S; Hampson, David J.

doi:10.1016/s0378-1135(99)00030-9

Cited by 83 publications

(77 citation statements)

References 33 publications

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“…1 Primer designing software b was used to design the following primers: forward: 5′-GTTCTTGGCCTGTAACTCCTCCTAT-3′; reverse: 5′-GCAACAATACCCATTCTTACAG-3′. A 620-bp region of the nox gene was then amplified using these primers.…”

Section: Brachyspira Speciationmentioning

confidence: 99%

Phenotypic and molecular characterization of a novel strongly hemolytic Brachyspira species, provisionally designated “Brachyspira hampsonii”

et al. 2012

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Abstract. Since 2007, outbreaks of severe bloody diarrhea and hemorrhagic colitis have been reported in the United States and Canada. Though the primary causative agent of swine dysentery is Brachyspira hyodysenteriae, which is strongly hemolytic, the current report describes the isolation of a novel strongly hemolytic Brachyspira sp. This novel Brachyspira sp. was identified from clinical submissions at the Minnesota Veterinary Diagnostic Laboratory, and 40 of such isolates were obtained from 22 clinical submissions representing 5 states. Isolates were confirmed to be different from any known Brachyspira sp. on the basis of phylogenetic analysis of nucleotide sequences of nox and 16S ribosomal RNA (rRNA) genes. Phylogenetic analyses grouped all isolates into 2 clades (clades I and II), and grouping patterns were similar for both nox and 16S rRNA gene sequence analyses. Phenotypically, all isolates were indole and hippurate negative, and enzymatic profiling indicated 2 types of profiles, irrespective of the phylogenetic grouping, differing only in the production of β-glucosidase. The results suggest that a potentially virulent new species of Brachyspira sp., provisionally named "Brachyspira hampsonii ", is circulating among swine herds in the United States.

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Section: Brachyspira Speciationmentioning

confidence: 99%

Phenotypic and molecular characterization of a novel strongly hemolytic Brachyspira species, provisionally designated “Brachyspira hampsonii”

et al. 2012

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“…PCR amplification was in 60 μl reaction mixtures with Taq DNA polymerase (Invitrogen) using the previously described primers and protocol (Atyeo et al, 1999;Townsend et al, 2005). Positive and negative controls were B. hyodysenteriae WA1 R and ultrapure water respectively.…”

Section: Nox Sequencingmentioning

confidence: 99%

“…Consequently other identification methods have been developed based on the sequence of conserved genes, notably those encoding 16S rRNA, 23S rRNA and NADH oxidase (nox). Analysis of nox sequences has emerged as a robust method for identification of Brachyspiraspecies as the gene is relatively conserved but also shows speciesspecific variation (Atyeo et al, 1999;Burrough et al, 2012;Chander et al, 2012;Rubin et al, 2013). A more discriminatory multilocus sequence typing (MLST) scheme analyzing the sequence of seven genes encoding "housekeeping" enzymes has been described for identification and typing of Brachyspira isolates (Råsbäck et al, 2007b), but it is laborious and has not been widely used apart from analysis of B. hyodysenteriae (La et al, 2009;Osorio et al, 2012) and other indole-positive species (Phillips et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Identification of weakly haemolytic Brachyspira isolates recovered from pigs with diarrhoea in Spain and Portugal and comparison with results from other countries

Osorio

Carvajal

Naharro

et al. 2013

Research in Veterinary Science

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Weakly haemolytic anaerobic intestinal spirochaetes of the genus Brachyspira are commonly identified based on species-specific gene sequences. Apart from the pathogenic Brachyspira pilosicoli, the distribution and disease associations of the other weakly haemolytic Brachyspira species in pigs have not been comprehensively investigated. In this study weakly haemolytic Brachyspira isolates (n = 67) from Spanish and Portuguese pigs with diarrhoea, negative in a routine diagnostic PCR for B. pilosicoli, were identified by sequencing their NADH oxidase genes (nox). Nearly half the isolates were identified as Brachyspira murdochii (n = 31; 46.3%). The others were Brachyspira innocens (n = 26; 38.8%), Brachyspira intermedia(n = 7; 10.4%), "Brachyspira pulli" (n = 1; 1.5%) and a potentially novel Brachyspira species (n = 2; 3%). Multilocus sequence typing (MLST) on a subset of 18 isolates confirmed their species designations, including the potential new species, and identified similarities to strains from other countries.

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“…A total of 30 different isolates, preferably from different herds, were selected for nox gene amplification to identify species belonging to genus Brachyspira and subsequent genomic sequencing, according to Chander et al (2012). The following primers were used (primer): Brachy nox F 5'-GTT CTT GCG CTG TAA CTC CTC CTA T-3') and Brachy nox R (5'-GCA ACA ATA CCC ATT CTT ACA G -3'), both of which are nox -specific and target a highly variable region of the gene (Atyeo et al 1999).…”

Section: Samplesmentioning

confidence: 99%

Minimum inhibitory concentration of Brazilian Brachyspira hyodysenteriae strains

et al. 2017

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The objectives of this study were to characterize Brachyspira hyodysenteriae isolates and to evaluate the antimicrobial susceptibility patterns of strains obtained from pigs in Brazil based on the minimal inhibitory concentration test (MIC). The MIC was performed for 22 B. hyodysenteriae isolates obtained from 2011 to 2013 using the following antimicrobial drugs: tylosin, tiamulin, valnemulin, doxycycline, lincomycin and tylvalosin. Outbreaks of swine dysentery were diagnosed based on clinical presentation, bacterial isolation, gross and microscopic lesions, duplex PCR for B. hyodysenteriae and B. pilosicoli and nox gene sequencing. All obtained MIC values were consistently higher or equal to the microbiological cut-off described in the literature. The MIC 90 values for the tested drugs were 8μg/ml for doxycycline, >4μg/ml for valnemulin, 8μg/ml for tiamulin, 32μg/ml for tylvalosin, >64μg/ml for lincomycin and >128μg/ml for tylosin. These results largely corroborate those reported in the literature. Tiamulin, doxycycline and tylvalosin showed the lowest MIC results. All of the samples subjected to phylogenetic analysis based on the nox gene sequence exhibited similar results, showing 100% identity to B. hyodysenteriae. This is the first study describing the MIC pattern of B. hyodysenteriae isolated in Brazil.

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Differentiation of Serpulina species by NADH oxidase gene (nox) sequence comparisons and nox-based polymerase chain reaction tests

Cited by 83 publications

References 33 publications

Phenotypic and molecular characterization of a novel strongly hemolytic Brachyspira species, provisionally designated “Brachyspira hampsonii”

Phenotypic and molecular characterization of a novel strongly hemolytic Brachyspira species, provisionally designated “Brachyspira hampsonii”

Identification of weakly haemolytic Brachyspira isolates recovered from pigs with diarrhoea in Spain and Portugal and comparison with results from other countries

Minimum inhibitory concentration of Brazilian Brachyspira hyodysenteriae strains

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