This study aimed to assess the presence of thermotolerant Campylobacter spp., as one of the most important foodborne zoonotic pathogens, in three shellfish species: mussels (Mytilus galloprovincialis), oysters (Ostrea edulis) and queen scallops (Aequipecten opercularis). The samples were collected from nine locations in the Istrian aquatory, Croatia. Isolation of Campylobacter was done according to standard ISO method, and species were identified using multiplex PCR. Isolates identified as C. jejuni and C. lari were genotyped using multilocus sequence typing (MLST) to determine the potential source of contamination. Among 108 examined samples of bivalve molluscs, mussels dominated and were the only ones found positive for the presence of Campylobacter (25.6%). In total, 19 C. lari and 1 C. jejuni strains were isolated. C. lari isolates found in this study belong to 13 sequence types (STs), and 9 of them are newly described in this paper. Two out of the four previously described C. lari STs that were found in this study were previously found in human stool. The only C. jejuni isolate was found to be sequence type 1268, which belongs to ST-1275 clonal complex that is almost exclusively found in seabirds and can sporadically cause infection in humans. Regarding the obtained results, introducing surveillance of thermotolerant Campylobacter in shellfish in the Republic of Croatia is advised as an improvement for public health safety.
Gulls are a group of seabirds distributed worldwide that are an important reservoir of Salmonella spp. Salmonellosis is the second most commonly reported gastrointestinal infection in humans, and understanding the role wild birds have in spreading Salmonella can help to improve the health of humans and domestic animals. The mobility and migration capacity of gulls makes them an interesting group for research given their potential role in spreading pathogens. This paper presents the diversity and prevalence of Salmonella spp. in different gull species caught at a landfill in Zagreb in the winter months over a nine-year period from 2014-2022. In total, 1083 cloacal swabs were sampled from six gull species: Black-headed Gull (Larus ridibundus), Yellow-legged Gull (L. michahellis), Caspian Gull (L. cachinnans), Common Gull (L. canus), Lesser Black-back Gull (L. fuscus) and Herring Gull (L. argentatus). The prevalence of Salmonella was 5.82%, and 16 Salmonella serotypes were identified; S. Typhimurium had the highest prevalence (47.62%) followed by S. Enteritidis (12.69%) and S. Infantis (9.52%). To date, 82 Salmonella serotypes have been isolated in research on gulls in Europe, with S. Typhimurium as the most common, followed by S. Agona and S. Enteritidis. In this study, we found three serotypes not previously reported in gulls, S. Yalding, S. Reading and one with the antigenic formula O:17; H:z10; H:e,n,x,z15 (IIIb).
Campylobacter lari is a thermotolerant bacterium that sporadically causes gastrointestinal diseases in humans and can be found in wildlife and the environment. C. lari is an understudied species, especially in wild birds such as gulls. Gulls are potentially good carriers of pathogens due to their opportunistic behavior and tendency to gather in large flocks. During winter and their breeding period, 1753 gulls were captured, and cloacal swabs were taken to be tested for the presence of C. lari. From isolated bacteria, the DNA was sequenced, and sequence types (ST) were determined. Sixty-four swabs were positive for C. lari, and from those, forty-three different STs were determined, of which thirty-one were newly described. The whole genome was sequenced for 43 random isolates, and the same isolates were tested for antimicrobial susceptibility using the broth microdilution method to compare them to WGS-derived antimicrobial-resistant isolates. All the tested strains were susceptible to erythromycin, gentamicin, and chloramphenicol, and all were resistant to ciprofloxacin. Resistance to ciprofloxacin was attributed to a gyrA_2 T86V mutation. Genes connected to possible beta-lactam resistance (blaOXA genes) were also detected.
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