Shiga-toxigenic Escherichia coli from animal food sources in Mauritius: Prevalence, serogroup diversity and virulence profiles.

Shiga-toxigenic Escherichia coli (STEC) are important human pathogens associated with diarrhea and in some cases haemorrhagic colitis. Contaminated food derived from cattle and wildlife species are often associated with disease outbreaks. In this study, we report the prevalence, serogroup diversity and virulence profiles of STEC strains derived from cattle, rusa deer and pig. Of the 422 samples analyzed, STEC were detected in 40% (80/200) of cattle, 27.0% (33/122) of deer and 13.0% (13/100) of pigs. STEC isolates belonged to 38 O-serogroups whereby 5.2% (24/462) of the isolates belonged to clinically important EHEC-7 serogroups: O26 (n = 2), O103 (n = 1), O145 (n = 3) and O157 (n = 18). Fourteen serogroups (O26, O51, O84, O91, O100, O104, O110, O117, O145, O146, O156, O157, O177 and ONT) displayed multiple virulence profiles. We also identified two serovars (O117 and O119) in deer which are not well-documented in epidemiological surveys. 73.7% (28/38) of recovered O-serogroups are known to be associated with serious human illnesses including haemolytic uremic syndrome (HUS) and bloody diarrhea. STEC isolates harboring single genotypes stx1, stx2, eae and hlyA accounted for 3.0% (14/462), 9.1% (42/462), 47.6% (220/462) and 1.7% (8/462) of all STEC isolates screened, respectively. Virulence combinations stx1 and stx2 were harboured by 1.3% of isolates while strains with genetic profiles eae/hlyA were the second most prevalent amongst STEC isolates. The full known virulent genotypes (stx2/eae, stx1/stx2/eae, stx1/stx2/hlyA and stx2/eae/hlyA) were present in 22 of the 462 STEC strains. A total of 10 different virulence patterns were recovered amongst animal species. Phylogeny of the gnd gene showed that amongst STEC strains, serovar O100 outlined the main cluster. Fourteen (n = 14) different sequence types (STs) were identified from a panel of twenty (n = 20) STEC isolates. One of the isolate (PG007B) possessed a unique ST (adk 10, fumC 693, gyrB 4, icd 1, mdh 8, purA 8, recA 2) that could not be assigned using MLST databases. None of the ST's recovered in deer were observed in domestic species. Our findings shows that food associated animals found on the tropical island of Mauritius carry a diversity of STEC strains with many serovars known to be associated with human disease. This report indicates that increased awareness, surveillance and hygienic attention at critical stages of the human food chain are warranted.

Molecular characterization of Salmonella isolates by REP-PCR and RAPD analysis.

Eighteen Salmonella isolates from both human and food (non-human) sources (fish, meat, and poultry) were characterized using conventional culture methods, biochemical, serological, and molecular analyses. REP-PCR and RAPD produced DNA profiles for differentiation purposes. Enterobacterial repetitive intergenic consensus (ERIC), repetitive extragenic palindronic (REP) and BOXAIR primers were selected for REP-PCR and two arbitrary primers, namely OPP-16 and OPS-11 were used for RAPD to generate DNA fingerprints from the Salmonella isolates. REP-PCR method showed greater discriminatory power in differentiating closely related strains of the related strains of Salmonella and produced more complex banding patterns as compared with RAPD. A dendogram was constructed with both sets of profiles using SPSS Version 13.0 computer software and showed that most human isolates were separately clustered from the non-human isolates. Two of the human isolates were closely related to some of the non-human isolates. A good correlation was also observed between the serogrouping of the O antigen and the molecular profiles obtained from REP-PCR and RAPD data of the Salmonella isolates. The results of a principal coordinate analysis (PCA) corresponded to the clustering in the dendrogram.

Genetic variants of Banana streak virus in Mauritius.

Genetic variations among isolates of Banana streak virus (BSV) were assessed using two sets of primers. The virus, found in banana accessions in Mauritius, was compared to a Nigerian isolate from cultivar Obino l'Ewai (BSOEV). On the basis of the observed size of amplicons, some Mauritius strains were different from l'Ewai BSOEV. Both Southern blot hybridization and the nucleotide sequences of the PCR products confirmed that they were of episomal BSV origin. An isolate of sugarcane bacilliform virus (SCBV) was found to be also very similar to the BSV isolated from banana samples. Nucleotide sequence analysis showed that even the same size PCR products had differing sequences. The dendrogram placed the isolates from Mauritius in a cluster separate from BSV and SCBV from other geographical locations.

RAPD-PCR Fingerprinting and Southern Analysis of Xanthomonas axonopodis pv. dieffenbachiae Strains Isolated from Different Aroid Hosts and Locations.

Anthurium blight, caused by Xanthomonas axonopodis pv. dieffenbachiae, is a systemic disease of Anthurium and other aroids. The aims of this work were to study the genetic diversity among X. axonopodis pv. dieffenbachiae strains and to identify, from the polymerase chain reaction (PCR) profiles, DNA probes that would be specific for the pathovar dieffenbachiae. Twenty-five X. axonopodis pv. dieffenbachiae strains, isolated from different hosts and geographical locations including Mauritius, were fingerprinted using the random amplified polymorphic DNA (RAPD)-PCR technique. The fingerprints were analyzed by the National Taxonomy System Software (NTSYS). The specificity of some of the RAPD fragments selected from PCR profiles was tested by Southern analyses of the PCR products. Ten arbitrary primers were chosen from an initial set of 111 decamers. Two hundred and nine RAPD markers were generated in eight individual DNA profiles. A correlation was found between the serotypes and the RAPD profiles for some groups of isolates. A possible link was also observed between the host range of the isolates tested and their RAPD profiles for strains isolated from Dieffenbachia and Philodendron. These results were confirmed by Southern analysis. Cluster analysis by the unweighted pair group method, arithmetic average (UPGMA) confirmed that the pathovar is genetically diverse with some strains that were clustered together showing similar host preferences. DNA probes with a potential use in molecular diagnostics of Anthurium blight were identified. This preliminary work could be used to develop PCR primers that will enable the sensitive detection of the pathogen in latently infected plants.

Serotyping and RAPD profiles of Salmonella enterica isolates from Mauritius.

AIMS: The genus Salmonella is a common agent of gastroenteritis in Mauritius, generating more cases of the disease during summer than during winter. The aims of this study were to assess the genetic diversity of isolates of Salmonella enterica by RAPD fingerprinting, and to establish the relationship between human and chicken isolates. METHODS: Twenty-six isolates were obtained from hospital laboratories and commercial poultry producers locally. RESULTS: The RAPD profiles, biochemical and serological analyses showed that two of the chicken isolates were mistakenly identified as Salmonella. The genetic diversity of the remaining 24 isolates (five chicken and 19 human), confirmed as Salmonella, was analysed using four arbitrary primers, OPA-10, OPR-03, OPI-06 and OPJ-09, chosen from an initial set of 10 decamers. Seventy RAPD markers were generated in four individual DNA profiles. SIGNIFICANCE AND IMPACT OF THE STUDY: Cluster analysis (UPGMA) performed using the NTSYS-pc V 1.8 computer software, confirmed that some strains of Salmonella isolated from chicken were genetically similar to those isolated from humans. Furthermore, a 1 kbp band amplified using primer OPA-10 was specific for the Salmonella genus as it was not amplified in any of the control bacteria.

Genetic polymorphism in Xanthomonas albilineans strains originating from 11 geographical locations, revealed by two DNA probes.

Two DNA fragments from Xanthomonas albilineans were used as probes to study the molecular diversity among strains of this pathogen. Two serologically distinct groups, serovars I and II, could be differentiated by hybridization to the probes. These probes, designated 830 and 838, were cloned after subtractive DNA hybridization of common sequences of Xanthomonas campestris pv. vasculorum from a serovar I strain of X. albilineans. They did not hybridize to the DNA of several other xanthomonads or to sugarcane DNA under the conditions of hybridization used. Faint bands were observed upon hybridization of probe 830 with one strain of X. campestris pv. phaseoli. The same banding patterns were obtained with a strain of X. albilineans from Burkina Faso and the serovar II strains of Mauritius. The serovar I strains from Mauritius and two other strains each from Reunion and South Africa had similar pattern.