Campylobacter spp. is a major cause of foodborne diseases in humans, particularly when transmitted by the handling or consumption of undercooked poultry meat. Most Campylobacter infections are self-limiting, but antimicrobial treatment (e.g., fluoroquinolones and macrolides) is necessary in severe or prolonged cases. The indiscriminate use of these drugs, both in clinical medicine and animal production, has a major impact on public health. The aim of the present study was to identify Campylobacter strains, isolated from turkey and broilers, using both PCR and the matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) methods to reveal the accuracy of identification, as well to evaluate the antimicrobial and genetic resistance of the investigated strains. MALDI-TOF and PCR methods were used to show differences, if any, in the specificity of that test. In this study, MALDI-TOF mass spectrometry gave the same results as multiplex PCR, in all cases. The highest rate of resistance (i.e., 100% of turkey and broiler strains) was detected against ciprofloxacin, whereas 58.1% of turkey and 78.6% of broiler strains were resistant to tetracycline. Multidrug-resistant isolates were not found in the study. All ciprofloxacin-resistant strains had a mutation in the gyrA gene, at the Thr-86 position. The presence of the tetO gene was found in 71% of turkey and in 100% of broiler strains. All resistant to tetracycline strains included tetO gene. Additionally, in five turkey and three broiler strains, susceptible to tetracycline, tetO gene was present. These results indicate the high prevalence of Campylobacter strains, which are phenotypically and genetically resistant to fluoroquinolones and tetracycline.
BackgroundSalmonella species are widespread in the environment, and occur in cattle, pigs, and birds, including poultry and free-living birds. In this study, we determined the occurrence of Salmonella in different wild bird species in Poland, focusing on five Salmonella serovars monitored in poultry by the European Union: Salmonella serovars Enteritidis, Typhimurium, Infantis, Virchow, and Hadar. We characterized their phenotypic and genetic variations.Isolates were classified into species and subspecies of the genus Salmonella with a polymerase chain reaction (PCR) assay. The prevalence of selected virulence genes (spvB, spiA, pagC, cdtB, msgA, invA, sipB, prgA, spaN, orgA, tolC, ironN, sitC, ipfC, sifA, sopB, and pefA) among the isolated strains was determined. We categorized all the Salmonella ser. Typhimurium strains with enterobacterial repetitive intergenic consensus (ERIC)-PCR.ResultsSixty-four Salmonella isolates were collected from 235 cloacal swabs, 699 fecal samples, and 66 tissue samples (6.4% of 1000 samples) taken from 40 different species of wild birds in Poland between September 2011 and August 2013. The largest numbers of isolates were collected from Eurasian siskin and greenfinch: 33.3% positive samples for both. The collected strains belonged to one of three Salmonella subspecies: enterica (81.25%), salamae (17.19%), or houtenae (1.56%). Eighteen strains belonged to Salmonella ser. Typhimurium (28.13%), one to ser. Infantis (1.56%), one to ser. Virchow (1.56%), and one to ser. Hadar (1.56%). All isolates contained spiA, msgA, invA, lpfC, and sifA genes; 94.45% of isolates also contained sitC and sopB genes. None of the Salmonella ser. Typhimurium strains contained the cdtB gene. The one Salmonella ser. Hadar strain contained all the tested genes, except spvB and pefA; the one Salmonella ser. Infantis strain contained all the tested genes, except tspvB, pefA, and cdtB; and the one Salmonella ser. Virchow strain contained all the tested genes, except spvB, pefA, cdtB, and tolC.The Salmonella ser. Typhimurium strains varied across the same host species, but similarity was observed among strains isolated from the same environment (e.g., the same bird feeder or the same lake).ConclusionsOur results confirm that some wild avian species are reservoirs for Salmonella serotypes, especially Salmonella ser. Typhimurium.
Salmonella enterica serovar Enteritidis has emerged during the last 20 years as the major causative agent of food-borne gastroenteritis in humans and as the major infectious agent on poultry farms, replacing Salmonella enterica serovar Typhimurium as the dominant pathogenic serovar. Because adhesion to gut tissues and colonization of the alimentary tract, mediated in large part by the FimH adhesins located on type 1 fimbriae, is an important stage in the pathogenesis of both serovars, the binding properties of the FimH adhesins from these two enteropathogens were compared. Salmonella Enteritidis FimH protein and the Salmonella Typhimurium low-adhesive variant of this adhesin were expressed in Escherichia coli and the recombinant proteins were analysed for their ability to bind glycoproteins carrying different oligomannosidic structures and different types of eukaryotic cells. In static binding assays (ELISA and Western blotting) both FimH proteins bound equally well to all three tested glycoproteins (RNase B, horseradish peroxidase and mannan-BSA). In addition, no differences were found in the binding specificity of the FimH proteins and intact cells of Salmonella Enteritidis and Salmonella Typhimurium to human colon carcinoma or bladder cancer cells. The presence of the same amino acid residues at positions 61 (glycine) and 118 (phenylalanine) and the similar binding properties of these two adhesins suggest that the newly described FimH protein of Salmonella Enteritidis represents the low-adhesive variant found in Salmonella Typhimurium. To study the binding specificity of Salmonella Enteritidis FimH protein further, direct kinetic analysis using surface plasmon resonance was performed. With this method it was found that Salmonella Enteritidis FimH adhesin bound with the highest K d value to high-mannose type N-glycans carried by RNase B; about 100 times lower K d values were obtained in the interactions with mannan-BSA and horseradish peroxidase.
Recombinant FimH adhesins of type 1 fimbriae from Salmonella enterica serovar Gallinarum biovars Gallinarum and Pullorum, in contrast to those of Salmonella enterica serovar Typhimurium, did not bind to high-mannose oligosaccharides or to human colon carcinoma HT-29 cells. However, mutated FimH proteins from biovar Gallinarum and biovar Pullorum, in which the isoleucine at position 78 was replaced by the threonine found in S. enterica serovar Typhimurium, bound well to glycoproteins carrying high-mannose oligosaccharides and colon carcinoma cells. The loss of sugar-binding properties by biovar Gallinarum and biovar Pullorum FimH adhesins, which are a part of the type 1 fimbriae, is most probably the result of a single T78I mutation, as was proven by site-directed mutagenesis of FimH proteins.
Campylobacter jejuni/coli infections are the leading cause of bacterial diarrheal illnesses in humans. Many epidemiological studies indicate that improperly prepared meat from chickens that carry a high load of Campylobacter in their intestinal tracts is the key source of human infections. LAB, mainly members of the Lactococcus and Lactobacillus genera, increasingly have been tested as vehicles for the delivery of heterologous bacterial or viral antigens to animal mucosal immune systems. Thus, the objective of this study was to isolate, identify, and characterize Lactobacillus spp. strains isolated from chickens bred in Poland. Their ability to decrease the level of bird gut colonization by C. jejuni strain was also analyzed. First, the influence of the different chicken rearing systems was evaluated, especially the effect of diets on the Lactobacillus species that colonize the gut of chickens. Next, selected strains were analyzed in terms of their anti‐Campylobacter activity in vitro; potential probiotic traits such as adhesion properties, bile and low pH tolerance; and their ability to grow on a defined carbon source. Given that improperly prepared chicken meat is the main source of human infection by Campylobacter, the selected strains were also assessed for their ability to inhibit Campylobacter colonization in the bird's intestine. These experiments revealed enormous physiological diversity among the Lactobacillus genus strains. Altogether, our results showed that L. plantarum strains isolated from the digestive tracts of chickens bred in Poland displayed some probiotic attributes in vitro and were able to decrease the level of bird gut colonization by C. jejuni strain. This suggests that they can be employed as vectors to deliver Campylobacter immunodominant proteins to the bird's immune system to strengthen the efficacy of in ovo vaccination.
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