Retail chicken giblets contaminated with extended-spectrum cephalosporin- and carbapenem-resistant Salmonella enterica carrying blaCMY-2.

Background and Aim: Chickens are considered as the main source of Salmonella, with infection potentially spreading to the public through outlets. The study aimed to investigate poultry shops for Salmonella enterica resistant to extended-spectrum cephalosporins-resistant (ESCR) and carbapenems-resistant (CR). Materials and Methods: Samples were collected from chicken giblets, water tanks, and workers at retail shops. Salmonella was isolated and serotyped; the presence of invA, stn, ompA, and ompF was determined using polymerase chain reaction (PCR). The isolates were tested for ESCR and CR by a disk-diffusion test; a confirmatory extended-spectrum ß-lactamase (ESBL) test was performed by combinational disk-diffusion test with clavulanic acid. The resistant isolates were screened for ESBL (blaTEM, blaSHV, blaCTX-M, and blaOXA-1), AmpC blaCMY-2, and carbapenemase (blaKPC, blaNDM, and blaOXA-48) genes using PCR. Results: S. enterica was isolated from chicken giblets (13/129) and the 13 isolates were ESCR. Based on the confirmatory ESBL test and CR, the 13 isolates were classified into the following resistance phenotypes: ESBL-producing and CR (n=4), ESBL-producing (n=1), non-ESBL-producing and CR (n=6), and non-ESBL-producing (n=2). All the five isolates with ESBL-producing phenotype carried predominantly blaTEM, blaSHV, and blaCMY-2. Regardless of being phenotypically CR, none of these isolates carried any of the tested carbapenemase genes. Surprisingly, the isolates with non-ESBL phenotype were found to carry blaTEM, blaSHV, and blaCMY-2. The blaKPC was present mainly in the isolates with non-ESBL and CR phenotypes. Interestingly, two isolates of the non-ESBL and CR phenotype showed resistance to cefepime, the fourth generation cephalosporins. Salmonella was also recovered from the water tanks (2/7) and the workers (2/16). The four isolates were ESCR and showed a non-ESBL-producing and CR phenotype; they harbored blaTEM, blaSHV, blaOXA-1, and blaKPC. The blaCMY-2 was found in one isolate from water and one from humans. All Salmonella isolates carried invA, stn, ompA, and ompF. Conclusion: Virulent ESCR S. enterica were identified in retail shops. The isolates carried blaCMY-2 and ESBL-genes, with a high proportion showing CR. Transmission of such strains to humans through food leads us to recommend regular inspection of retail outlets for antibiotic-resistant bacteria.

Sequencing and phylogenetic analysis of the stn gene of Salmonella species isolated from different environmental sources at Lake Qarun protectorate: The role of migratory birds and public health importance.

BACKGROUND AND AIM: Salmonella causes most foodborne bacterial illnesses worldwide. It is found in various hosts, including pets, farm animals, and wild animals, as well as the environment. This study aimed to examine the epidemiological relationship between Salmonella isolates from aquatic environments and those from other avian hosts. MATERIALS AND METHODS: The study examined 12 water samples, 210 aquatic animals, and 45 migratory aquatic bird samples collected from the protected area of Lake Qarun in El-Fayoum Governorate, Egypt, during migration seasons from different waterfowl migration areas (from October 2018 to January 2019). In addition, 45 fecal samples from domestic chickens were collected from the same geographic location from poultry farms. Bacteriological examination and polymerase chain reaction assay of two virulence genes (i.e., invA and stn) were performed to isolate and identify Salmonella. RESULTS: Salmonella was isolated from 58.3% (7/12) of Lake Qarun water samples, 13.3% (6/45) of migratory waterfowl, 6.6% of (3/45) of chickens (Gallus gallus domesticus), and 4.3% (3/70) of fish and pooled brine shrimp. In migratory aquatic bird species that were sampled, Salmonella were isolated from 23.1% (3/13) of Eurasian coot (Fulica atra), 12.5%, (1/8) of green-winged teal (Anas cardolinesis), 10% (2/20) of northern shoveler (Spatula clypeata), and 0% (0/4) of mallard duck (Anas platyrhynchos). In 35 Tilapia, Salmonella was isolated by (8.6%) 5.7% of external surfaces, 2.85% from the intestine, and 0% from the muscle. No Salmonella was isolated from the 175 brine shrimp samples. Phylogenetic analysis using the stn genes of Salmonella isolated from the aquatic environment, migratory aquatic birds, and chicken showed a strong association between these isolates. In addition, a higher nucleotide identity percentage was observed between the sequences recovered from migratory aquatic birds and Lake Qarun water samples. CONCLUSION: Salmonella distribution was confirmed through migratory aquatic birds, based on our phylogeny tree analysis, Salmonella considered a likely carrier of zoonotic bacterial pathogens. Furthermore, the close relationship between chicken and fish sequences highlights the scenarios of using chicken manure in fish farms and its public health implications. The presence of Salmonella in different environmental sources spotlights the urgent need to control and break down its epidemiological cycle.

Extended-spectrum ß-lactamase-producing Salmonella serovars among healthy and diseased chickens and their public health implication.

OBJECTIVES: This study investigated the occurrence of extended-spectrum ß-lactamase (ESBL)-producing Salmonella and the associated virulence genes among farmed chickens. METHODS: Cloacal swab samples were collected from apparently healthy and diseased chickens and were cultured for Salmonella using conventional methods. The isolates were serotyped using slide agglutination tests and were examined by polymerase chain reaction (PCR) for the virulence genes invA, stn, svpC and pefA and the outer membrane protein-encoding genes ompA and ompF. Screening for ESBL resistance was performed using the disk-diffusion test, the combinational-disk test with clavulanic acid, and multiplex PCR for blaTEM, blaSHV, blaCTX-M and blaOXA. The presence of the AmpC blaCMy-2 was tested among the ESBL-negative isolates by uniplex PCR. The resistant isolates were partially sequenced based on the stn gene. RESULTS: The Salmonella isolation rate was 3.4% (6/175) from healthy and 11.1% (14/126) from diseased chickens. The 20 isolates belong to serotypes with public health significance like Typhimurium, Kentucky and Infantis. All the isolates possess invA, stn, svpC and ompF genes; 16 isolates harboured ompA, and one carried pefA. Of the 20 isolates, 19 were resistant to more than one antibiotic. Of these 19 isolates, 16 were ESBL-producing with the majority carrying blaTEM and blaSHV genes. The four ESBL-negative isolates carried blaCMY-2. Partial-stn-sequencing of the isolates revealed a high genetic relatedness to Salmonella strains from patients in Egypt and Asia. CONCLUSIONS: Virulent ESBL-producing Salmonella was isolated from healthy and diseased chickens; the strains have a close relationship to human strains, posing a public health threat.

Occurrence of Clostridium perfringens Types A, E, and C in Fresh Fish and Its Public Health Significance.

Fish remains among the most traded of food commodities, and Egypt is one of the emerging countries being recognized as an important world fish exporter. Clostridium perfringens is an important foodborne pathogen to consider in fish trade, as it has been implicated as the causative organism of two fish outbreaks. The aim of the present study was to investigate the occurrence and toxin diversity of C. perfringens associated with fresh and canned fish and to examine the public health significance of C. perfringens infection in fish. Isolation and identification of C. perfringens showed a significantly higher prevalence of the bacterium in fresh fish collected from aquaculture (54.5%) and from markets (71%) as well as in humans in contact with fish (63%) compared with water used for keeping fresh fish (27.3%) and water used in canned fish (17.8%). The isolation level was significantly higher in samples from the external surface of fresh fish (31.8% in aquaculture, 45.6% in markets) than from the intestinal contents of the same fish (9.1% in aquaculture, 6.7% in markets). Thus, markets represent a risk factor for contamination of the external surface of fish from the surrounding environment. Genotyping of the C. perfringens-positive isolates by using multiplex PCR revealed that type A enterotoxin-negative (CPE(-)) is the predominant strain among fish (fresh and canned), humans, and water in contact with fresh fish. Interestingly, C. perfringens types A enterotoxin-positive (CPE(+)) and C were found only in fresh fish, and these two strains have great health importance in humans. Strikingly, C. perfringens type E strain was detected for the first time in fish, humans, and water in contact with fresh fish. Our results demonstrate for the first time that fish act as a reservoir for C. perfringens, particularly for types A CPE(+), C, and E. The external surface of fish represents a vehicle for contamination of fish from the surrounding environment as well as a source of infection of humans, thereby representing a public health hazard.