Comparison of Antibiofilm Activity of Pseudomonas aeruginosa Phages on Isolates from Wounds of Diabetic and Non-Diabetic Patients.

The persistence of organisms as biofilms and the increase in antimicrobial resistance has raised the need for alternative strategies. The study objective was to compare the ability of isolated bacteriophages to remove in vitro biofilms formed by Pseudomonas aeruginosa isolated from the environment with those isolated from diabetic and non-diabetic wounds. P. aeruginosa were isolated from clinical and environmental sites, and antimicrobial susceptibility was tested. Bacteriophages were isolated and characterized based on plaque morphology and host range. A reduction in the viable count assayed the lytic ability of candidate phages. The crystal violet method was used to determine the residual biofilm after 24 h of phage treatment on 72-h-old biofilms. The statistical significance of phage treatment was tested by one-way ANOVA. Of 35 clinical isolates, 17 showed resistance to 1 antibiotic at least, and 7 were multidrug resistant. Nineteen environmental isolates and 11 clinical isolates were drug-sensitive. Nine phages showed 91.2% host coverage, including multidrug-resistant isolates. Phages eradicated 85% of biofilms formed by environmental isolates compared to 58% of biofilms of diabetic isolates and 56% of biofilms of non-diabetic isolates. Clinical isolates are susceptible to phage infection in planktonic form. Biofilms of P. aeruginosa isolated from diabetic wounds and non-diabetic wounds resist removal by phages compared to biofilms formed by environmental isolates. All phages were efficient in dispersing PAO1 biofilms. However, there was a significant difference in their ability to disperse PAO1 biofilms across the different surfaces tested. Partial eradication of biofilm by phages can aid in complementing antibiotics that are unable to penetrate biofilms in a clinical set-up.

Biocontrol of Escherichia coli and Salmonella in poultry meat using phage cocktail.

Background: Despite advances in food management techniques, foodborne illness remains a major concern. Contamination of Salmonell a and Escherichia coli pathogens, especially in the poultry sector, is responsible for salmonellosis and other gastrointestinal illness, leading to millions of deaths worldwide. Overuse of antibiotics and other chemical treatments have further increased the emergence of antibiotic resistant bacteria. Aims: This study aimed to study the efficacy of phages cocktail to reduce the load of E. coli and Samlonella spiked on poultry meat. Methods: In this study, a broad spectrum cocktail of phages was used to lyse E. coli and Salmonella spiked on chicken meat. Results: Based on the result of the CFU drop assay, phages like E. coli 153T 3ii and Salmonella 191(3) were selected. Phage concentration of 0.01 MOI showed a reduction in E. coli and Salmonella count to 6 h and 2 h, respectively. Further, phages were tested on the surface of chicken meat. E. coli showed a 90% reduction up to 4 h, whereas Salmonella showed a 90% reduction up to 6 h. When phages were treated in combination, a significant reduction of up to 12 h was found with Salmonella phage, showing better antimicrobial activity. Conclusion: The suitable concentration of a specific phage or phage cocktail can significantly reduce the bacterial count on chicken meat. Phage mediated biocontrol can be used as an alternative approach to eliminate enteric pathogens in the poultry industry.

Application of bacteriophages to control Salmonella Enteritidis in raw eggs.

BACKGROUND: Salmonella, a bacterial genus of more than 2500 serotypes is considered as the most significant foodborne pathogen causing infections in humans and animals. Increased antimicrobial resistance and persistence of antimicrobial residues in food matrices warrants the need for alternative infection management strategies. AIMS: The present study aimed to isolate and evaluate the lytic activity of bacteriophage against Salmonella. METHODS: Twenty-eight Salmonella isolates obtained from the poultry sources were screened for antibiotic sensitivity. Poultry slaughterhouse wastewater was used for the isolation of phage. Host range and random amplified polymorphic DNA (RAPD) are vital tools used for differentiating the phage. RESULTS: The isolates showed a high degree of resistance to nalidixic acid (71%), tetracycline (71%), nitrofurantoin (50%), and ampicillin (43%). Five lytic phages are specific for Salmonella spp. were isolated and characterized by RAPD. In the colony forming unit (CFU) reduction assay, the highest activity of phage was observed at 0.01 multiplicity of infection (MOI) within 2 h after the addition of phage. PSE5 at 0.01 MOI was administered to Salmonella Enteritidis seeded on the surface of the chicken egg by immersion method. The results indicated that administration of phage reduced recoverable Salmonella by 2 × 106 CFU/ml relative to the phage-excluded control. CONCLUSION: The results presented here suggested the application of the bacteriophage treatment has the potential to be used as an alternative strategy to prevent Salmonella infection in poultry farms to prevent vertical transmission of the pathogen.

Incidence of subclinical mastitis and prevalence of major mastitis pathogens in organized farms and unorganized sectors.

Subclinical mastitis (SCM) represents a major proportion of the burden of mastitis. Determining somatic cell count (SCC) and electrical conductivity (EC) of milk are useful approaches to detect SCM. In order to correlate grades of SCM with the load of five major mastitis pathogens, 246 milk samples from a handful of organized and unorganized sectors were screened. SCC (>5 × 10(5)/mL) and EC (>6.5 mS/cm) identified 110 (45 %) and 153 (62 %) samples, respectively, to be from SCM cases. Randomly selected SCM-negative samples as well as 186 samples positive by either SCC or EC were then evaluated for isolation of five major mastitis-associated bacteria. Of the 323 isolates obtained, 95 each were S. aureus and coagulase-negative staphylococci (CoNS), 48 were E. coli and 85 were streptococci. There was no association between the distribution of organisms and (a) the different groups of SCC, or (b) organised farms and unorganised sectors. By contrast, there was a significant difference in the distribution of CoNS, and not other species, between organized farms and unorganized sectors. In summary, bacteria were isolated irrespective of the density of somatic cells or the type of farm setting, and the frequency of isolation of CoNS was higher with organized farms. These results suggest the requirement for fine tuning SCC and EC limits and the higher probability for CoNS to be associated with SCM in organized diary sectors, and have implications for the identification, management and control of mastitis in India.