Phenotypic and genotypic characterization of resistance and virulence in Pseudomonas aeruginosa isolated from poultry farms in Egypt using whole genome sequencing.

Pseudomonas aeruginosa (P. aeruginosa) is an ESKAPE pathogen that can quickly develop resistance to most antibiotics. This bacterium is a zoonotic pathogen that can be found in humans, animals, foods, and environmental samples, making it a One-Health concern. P. aeruginosa threatens the poultry industry in Egypt, leading to significant economic losses. However, the investigation of this bacterium using NGS technology is nearly non-existent in Egypt. In this study, 38 isolates obtained from broiler farms of the Delta region were phenotypically investigated, and their genomes were characterized using whole genome sequencing (WGS). The study found that 100% of the isolates were resistant to fosfomycin and harbored the fosA gene. They were also resistant to trimethoprim/sulfamethoxazole, although only one isolate harbored the sul1 gene. Non-susceptibility (resistant, susceptible with increased dose) of colistin was observed in all isolates. WGS analysis revealed a high level of diversity between isolates, and MLST analysis allocated the 38 P. aeruginosa isolates into 11 distinct sequence types. The most predominant sequence type was ST267, found in 13 isolates, followed by ST1395 in 8 isolates. The isolates were susceptible to almost all tested antibiotics carrying only few different antimicrobial resistance (AMR) genes. Various AMR genes that confer resistance mainly to ß-lactam, aminoglycoside, sulfonamide, and phenicol compounds were identified. Additionally, several virulence associated genes were found without any significant differences in number and distribution among isolates. The majority of the virulence genes was identified in almost all isolates. The fact that P. aeruginosa, which harbors several AMR and virulence-associated factors, is present in poultry farms is alarming and threatens public health. The misuse of antimicrobial compounds in poultry farms plays a significant role in resistance development. Thus, increasing awareness and implementing strict veterinary regulations to guide the use of veterinary antibiotics is required to reduce health and environmental risks. Further studies from a One-Health perspective using WGS are necessary to trace the potential transmission routes of resistance between animals and humans and clarify resistance mechanisms.

Protective Impact of Chitosan Film Loaded Oregano and Thyme Essential Oil on the Microbial Profile and Quality Attributes of Beef Meat.

Edible films and essential oil (EO) systems have the potency to enhance the microbial quality and shelf life of food. This investigation aimed to evaluate the efficacy of chitosan films including essential oils against spoilage bacteria and foodborne pathogens associated with meat. Antimicrobial activity (in vitro and in vivo) of chitosan films (CH) incorporated with oregano oil (OO) and thyme oil (TO) at 0.5 and 1% was done against spoilage bacteria and foodborne pathogens, compared to the control sample and CH alone. Preliminary experiments (in vitro) showed that the 1% OO and TO were more active against Staphylococcus aureus compared to Escherichia coli O157:H7 and Salmonella Typhimurium. In in vivo studies, CH containing OO and TO effectively inhibited the three foodborne pathogens and spoilage bacteria linked with packed beef meat which was kept at 4 °C/30 days compared to the control. The total phenolic content of the EOs was 201.52 mg GAE L-1 in thyme and 187.64 mg GAE L-1 in oregano. The antioxidant activity of thyme oil was higher than oregano oil. The results demonstrated that the shelf life of meat including CH with EOs was prolonged ~10 days compared to CH alone. Additionally, CH-OO and CH-TO have improved the sensory acceptability until 25 days, compared to the control. Results revealed that edible films made of chitosan and containing EOs improved the quality parameters and safety attributes of refrigerated or fresh meat.

Efflux Pump Inhibitors, Alpha-Tocopherol and Aspirin: Role in Campylobacter jejuni and Campylobacter coli Fluoroquinolone Resistance.

This study aimed to investigate how efflux pump activity contributes to high fluoroquinolone (FQ) resistance in Campylobacter jejuni and Campylobacter coli isolates and to evaluate the modulatory effects of α-tocopherol and aspirin on FQ phenotypic resistance profiles. Minimum inhibitory concentration (MIC) values were obtained for different FQ agents following exposure to different efflux pump inhibitors (EPIs), including PaßN (50 µg/mL), which targets the cmeABC efflux system, and chlorpromazine (45 µg/mL) and verapamil (120 µg/mL), which target the MFS efflux system. The modulatory effects of aspirin (100 and 200 µg/mL) and α-tocopherol (4 and 10 µg/mL) on FQ resistance profiles were examined. PaßN had no effect on the MIC values of all FQ agents, while MFS efflux system inhibitors reduced the resistance level of different FQ agents and achieved an effect nearly comparable with that of α-tocopherol (10 µg/mL). Aspirin exerted a dose-dependent excitatory effect on phenotypic resistance profiles, and this may raise concerns about its usage in both veterinary and clinical settings. While an efflux system other than cmeABC may play a role in FQ resistance in Campylobacter species, lipophilic substances may represent a new approach for controlling efflux pump activities.

ERIC-PCR Genotyping of Some Campylobacter jejuni Isolates of Chicken and Human Origin in Egypt.

The public health importance of the genus Campylobacter is attributed to several species causing diarrhea in consumers. Poultry and their meat are considered the most important sources of human campylobacteriosis. In this study, 287 samples from chicken (131 cloacal swabs, 39 chicken skin, 78 chicken meat, and 39 cecal parts) obtained from retail outlets as well as 246 stool swabs from gastroenteritis patients were examined. A representative number of the biochemically identified Campylobacter jejuni isolates were identified by real-time PCR, confirming the identification of the isolates as C. jejuni. Genotyping of the examined isolates (n = 31) by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) revealed a high discriminatory index of ERIC-PCR (D = 0.948), dividing C. jejuni isolates of chicken and human origins into 18 profiles and four clusters. The 18 profiles obtained indicated the heterogeneity of C. jejuni. Dendrogram analysis showed that four clusters were generated; all human isolates fell into clusters I and III. These observations further support the existence of a genetic relationship between human and poultry isolates examined in the present study. In conclusion, the results obtained support the speculation that poultry and poultry meat have an important role as sources of infection in the acquisition of Campylobacter infection in humans.