Biofilm-forming ability, antibiotic resistance and phylogeny of Escherichia coli isolated from extra intestinal infections of humans, dogs, and chickens.

Escherichia coli (E. coli) causes various infections in humans and animals. The biofilm-forming ability of E. coli has increased antimicrobial resistance and capacity to cause recurrent and chronic infections. This study determined the biofilm-forming ability of E. coli isolated from extraintestinal infections of humans, chickens, and dogs in relation to the phylogroup, type of infection, and antibiotic resistance. Isolates from chickens showed significantly higher biofilm-forming ability compared to those causing urinary tract infections in humans (p = 0.0001). Further, isolates belonging to phylogroup B1 displayed a higher likelihood to form biofilms. Resistance to ciprofloxacin and trimethoprim-sulfamethoxazole was positively correlated with biofilm-forming ability. Harbouring plasmid-mediated quinolone resistance gene, qnrS was also positively correlated with biofilm formation. This study provides insight into factors such as phylogroup and the type of infections that could enhance biofilm formation, as well as genotypic and phenotypic antibiotic resistance that could correlate with the ability to form biofilms.

Analysis of risk factors associated with recurrence of canine babesiosis caused by Babesia gibsoni.

Canine babesiosis due to Babesia gibsoni (B. gibsoni) displays severe clinical manifestations. Recurrence of babesiosis after anti-babesial treatment is observable in over 10 % of the patients. The present study ascertains the risk factors and cumulative incidence of recurrence of canine babesiosis. For a sample of 145 dogs diagnosed with acute babesiosis, the following parameters were assessed over a period of 16 weeks: haematological parameters, status of anaemia, platelet count, total WBC count, haemoglobin concentration and RBC count, concurrent haemoparasitism, and secondary immune mediated haemolytic anaemia (IMHA). Patient demographics such as age, breed, sex were also recorded. The potential risk factors were statistically evaluated by the cumulative incidence function and the Kaplan-Meier method. The recurrent infections were observed in 11.8 % of the study sample. The following factors were found to associate with increased risk of recurrence: Rottweiler breed (CIR 21.8 % ± 6.9 %; p < 0.05), secondary IMHA (CIR 28.7 % ± 11.3 %; p < 0.05), RBC counts < 2 × 106/µl on the day of diagnosis (CIR 16 % ± 4.6 %; p < 0.05), and persistent anaemia over 20 days post treatment (CIR 29.14 ± 7.9 %; p < 0.001). Dogs with concurrent haemoparasitic infections were predicted to have a fatal outcome in the survival analysis (disease related mortalities 25 % ± 13 %; p < 0.001). According to the findings, veterinarians need to pay attention to Rottweiler breed, dogs with secondary IMHA, concurrent haemoparasitism, low RBC counts on diagnosis and those with persistent anaemia to reduce the risk of relapse.

Risk, prognosis and causality of parvo viral enteritis in dogs in Sri Lanka.

We studied the causal variation, prognosis and risk factors of parvo viral enteritis using both molecular and statistical analysis. A multivariate factor analysis was performed to see the effect of the breed, age, vaccination status and the leukocyte counts on disease severity and survival. Twenty PCR amplicons (partial VP2 gene) from vaccinated dogs were sequenced to identify the variants. A total of 109 samples were positive for parvo viral DNA, 71 of which were from dogs who have received at least one dose of CPV-2 vaccine. Over 90% were leukopenic on 2nd and 3rd day of diarrhoea. Age < 6 months was negatively correlated with leukocyte count and the leukocyte count was negatively correlated with survival. Dobermans and Labradors showed significant negative correlation with survival. All variants of parvovirus affected dogs with the 2c being the predominant (2c = 12, 2a = 7, 2b = 1). All 20 strains harboured three additional amino acid substitutions (i.e. Phe267Tyr, Ser297Ala and Tyr324Ile) and warrant further studies on potential changes of the antigenicity of the virus.

Population structure and virulence content of avian pathogenic Escherichia coli isolated from outbreaks in Sri Lanka.

Avian pathogenic Escherichia coli (APEC) causes economically significant infections in poultry. The genetic diversity of APEC and phylogenetic relationships within and between APEC and other pathogenic E. coli are not yet well understood. We used multilocus sequence typing (MLST), PCR-based phylogrouping and virulence genotyping to analyse 75 avian E. coli strains, including 55 isolated from outbreaks of colisepticaemia and 20 from healthy chickens. Isolates were collected from 42 commercial layer and broiler chicken farms in Sri Lanka. MLST identified 61 sequence types (ST) with 44 being novel. The most frequent ST, ST48, was represented by only six isolates followed by ST117 with four isolates. Phylogenetic clusters based on MLST sequences were mostly comparable to phylogrouping by PCR and MLST further differentiated phylogroups B1 and D into two subgroups. Genotyping of 16 APEC associated virulence genes found that 27 of the clinical isolates and one isolate from a healthy chicken belonged to highly virulent genotype according to previously established classification schemes. We found that a combination of four genes, ompT, hlyF, iroN and papC, gave a comparable prediction to that of using five and nine genes by other studies. Four STs (ST10, ST48, ST117 and ST2016) contained APEC isolates from this study and human UPEC isolates reported by others, suggesting that these STs are potentially zoonotic. Our results enhanced the understanding of APEC population structure and virulence association.

Occurrence of nontuberculous mycobacteria in aquatic sources of Sri Lanka.

Nontuberculous mycobacteria (NTM) have been reported to cause opportunistic infections with increasing frequency, especially in immunocompromised patients. Water plays a major role in the epidemiology of nontuberculous mycobacterial infection in humans, as it is one of the natural sources for transmission of this group of organisms. The current study focused on determining the occurrence of NTM in different aquatic sources of Sri Lanka by using phenotypic tests and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the rpoß gene. Of 290 water samples, 45 (15%) were positive for NTM on culture. The percentage of mycobacteria identified at species level by phenotypic tests and PCR-RFLP analysis were 44% (20/45) and 73% (33/45), respectively. The frequency of isolation of mycobacteria from aquarium water, surface water, ground water and chlorinated water were 29% (20/70), 26% (20/76), 5% (4/76) and 1% (1/68), respectively. Eleven different NTM species were identified by PCR-RFLP analysis. M. fortuitum type I was the most frequently isolated species from all the four water sources. The current study suggests that water is an environmental source harboring NTM, a potential public health hazard especially for those with immunodeficiency.

Distribution of lipopolysaccharide core types among avian pathogenic Escherichia coli in relation to the major phylogenetic groups.

Five distinct lipopolysaccharide (LPS) core types, namely R1-R4 and K12 have been identified in Escherichia coli. The aims of this study were to determine, primarily by means of PCR, the distribution of those oligosaccharide core types among avian pathogenic E. coli and their relationship to phylogenetic groups. To identify putative avian pathogenic E. coli, serum resistance and the presence of three virulence genes encoding temperature sensitive haemagglutinin (tsh), increased serum survival (iss) and colicin V (cvaC) were determined. Of the 143 clinical isolates examined 62% possessed the R1 core, 22% were R3, 13% were R4 and 3% were R2. Fifty commensal isolates consisted of 58% with R1 core, 38% with R3 core, 4% with R4 core, and none with R2. None of the isolates were of K12 core type. The distribution of core oligosaccharide types in clinical and commensal isolates were not statistically significant (P=0.51). Three genes, tsh, iss and cvaC were found in E. coli of all four core types. The genes tsh (P<0.001) and iss (P=0.03412) were significantly associated with the R4 core oligosaccharide type. The isolates containing R4 core type LPS were mainly confined to phylogenetic group D. The widespread R1 core type showed less ability to possess virulence genes and 83% were in the phylogenetic group A. Results of this study indicated that E. coli with R1, R2, R3 and R4 were important in causing infections in chickens and further, the E. coli with R4 core type were less common among commensals, possessed more virulence genes and were related to phylogenetic groups pathogenic for poultry.