The relationship between bacterial sources and genotype to the antimicrobial resistance pattern of Burkholderia pseudomallei.

BACKGROUND: Melioidosis is a fatal emerging infectious disease of both man and animal caused by bacteria Burkholderia pseudomallei. Variations were suggested to have existed among the different B. pseudomallei clinical strains/genotypes which may implicate bacterial susceptibility and resistance toward antibiotics. AIM: This study was designed to determine whether the phenotypic antibiotic resistance pattern of B. pseudomallei is associated with the source of isolates and the genotype. MATERIALS AND METHODS: A collection of 111 B. pseudomallei isolates from veterinary cases of melioidosis and the environments (soil and water) were obtained from stock cultures of previous studies and were phylogenetically characterized by multilocus sequence typing (ST). The susceptibility to five antibiotics, namely meropenem (MEM), imipenem, ceftazidime (CAZ), cotrimoxazole (SXT), and co-amoxiclav (AMC), recommended in both acute and eradication phases of melioidosis treatment were tested using minimum inhibitory concentration antibiotics susceptibility test. RESULTS: Majority of isolates were susceptible to all antibiotics tested while few resistant strains to MEM, SXT, CAZ, and AMC were observed. Statistically significant association was found between resistance to MEM and the veterinary clinical isolates (p<0.05). The likelihood of resistance to MEM was significantly higher among the novel ST 1130 isolates found in veterinary cases as compared to others. CONCLUSION: The resistance to MEM and SXT appeared to be higher among veterinary isolates, and the novel ST 1130 was more likely to be resistant to MEM as compared to others.

Phylogenetic Diversity of Burkholderia pseudomallei isolated from veterinary cases and the environments in Peninsular Malaysia.

This study was designed to determine the genotype and the phylogeny of Burkholderia pseudomallei isolated from veterinary cases and from the animal environments in Peninsular Malaysia. The Malaysian B. pseudomallei population were then compared to those found elsewhere. A total of 113 isolates from veterinary cases (35) and the environment (56 from soil and 22 from water) were characterized using multilocus sequence typing (MLST). Two novel alleles, allele 97 and 69 of the gene locus ace and lepA respectively were recovered. Isolates were resolved into 12 distinct sequence types (STs) out of which five were novel, namely ST1130, ST1131, ST1338, ST1339 and ST1367. The isolates from veterinary cases co-clustered with those from the environment. B. pseudomallei isolates in this study were highly clonal and have descended from a common ancestor clonal complex (CC) 48 found in Southeast Asia. This study shows that veterinary case isolates are often caused by similar STs, with similar populations found in the direct animal environment and those previously reported to cause human infections in Malaysia and elsewhere. Isolates of B. pseudomallei from human infections have been given more attention, with a comparatively lower focus on isolates from animals and the farm environment. This study highlighted the genotype and phylogeny of B. pseudomallei isolated from animals and the environment and their relations to the isolates from human cases reported in Malaysia and elsewhere. Most STs reported in this study, from veterinary cases and animal environment are similar to those previously reported as causing human infections in Malaysia and elsewhere. Therefore, even though direct zoonosis is uncommon, monitoring melioidosis occurrences in animals can provide insights on the bacterial strains infecting humans.

Intestinal spirochaetes (Brachyspira spp.) colonizing flocks of layer and breeder chickens in Malaysia.

Avian intestinal spirochaetosis causes problems including delayed onset of lay and wet litter in adult chickens, and results from colonization of the caecae/rectum with pathogenic intestinal spirochaetes (genus Brachyspira). Because avian intestinal spirochaetosis has not previously been studied in South East Asia, this investigation was undertaken in Malaysia. Faecal samples were collected from 25 farms and a questionnaire was administered. Brachyspira species were detected by polymerase chain reaction in 198 of 500 (39%) faecal samples from 20 (80%) farms, including 16 (94%) layer and four (50%) breeder farms. Pathogenic Brachyspira pilosicoli was identified in five (29%) layer and two (25%) breeder farms whilst pathogenic Brachyspira intermedia was detected in nine (53%) layer and one (12.5%) of the breeder farms. Twelve (80%) layer farms had egg production problems and 11 (92%) were positive for Brachyspira: three (25%) for B. pilosicoli and six (50%) for B. intermedia. Of three breeder farms with egg production problems, one was colonized with B. pilosicoli. Three of ten layer farms with wet litter were positive for B. pilosicoli and six for B. intermedia. Of four breeder farms with wet litter, one was colonized with B. pilosicoli and one with B. intermedia. No significant associations were found between colonization and reduced egg production or wet litter, perhaps because so many flocks were colonized. A significant association (P = 0.041) occurred between a high prevalence of colonization and faecal staining of eggs. There were significant positive associations between open-sided housing (P = 0.006), and flocks aged >40 weeks (P < 0.001) and colonization by pathogenic species.

Brachyspira intermedia strain diversity and relationships to the other indole-positive Brachyspira species.

The aims of this study were to use multilocus sequence typing (MLST) to (i) investigate the population structure, diversity and molecular epidemiology of the weakly haemolytic anaerobic intestinal spirochaete Brachyspira intermedia, and (ii) determine the relationship of the species to the other two indole-positive but strongly haemolytic Brachyspira species--B. hyodysenteriae and "B. suanatina". Seventy-seven B. intermedia isolates from pigs and chickens were analysed, with the nucleotide sequences of seven conserved genomic loci examined for each. B. intermedia was genetically diverse, with the 77 isolates being divided into 71 sequence types (STs) and 64 amino acid types (AATs). Many distinct groups of B. intermedia isolates were identified, with some isolates being separated from others by large genetic distances. Although both pig and chicken isolates were found in most groups, suggesting that cross-species transmission of such isolates may occur, some isolates from pigs were located in small groups that did not include chicken isolates, and vice versa. Eight clonal complexes (Cc) of STs were identified by e-Burst analysis. The Ccs contained between 2 and 5 STs, and between 2 and 9 isolates. Five Ccs contained multiple isolates from the same farms, collected at the same time, indicating the existence of ongoing minor genetic change amongst isolates at the farm level. On the other hand, isolates with quite different STs also were found amongst multiple isolates collected from some farms. By comparison with the much more restricted diversity observed for 111 isolates of B. hyodysenteriae, and 4 isolates of "B. suanatina", it is difficult to justify including all weakly haemolytic indole-positive Brachyspira isolates in the single species B. intermedia.