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•Caspian gulls are involved in the circulation of the AMR Escherichia.•Closely related Escherichia strains were detected in countries frequented by gulls.•The prevalence of AMR can ...fluctuate in gulls and drop significantly in time.•Several Escherichia STs exhibited persistence within the gulls' colony in different years.•Clinically important traits may be not relevant for STs environmental persistence.
This study is focused on Escherichia spp. isolates resistant to critically important antibiotics (cefotaxime, ciprofloxacin and colistin) among Caspian gull’s (Larus cachinnans) chicks nesting in the Nove Mlyny Water Reservoir, Czech Republic. The prevalence of antimicrobial resistance (AMR) in bacteria within wild birds is commonly evaluated using a single sampling event, capturing only a brief and momentary snapshot at a particular location. Therefore, the Caspian gulls in our study were sampled in May 2018 (n = 72) and May 2019 (n = 45), and a water sample was taken from the reservoir (2019). We obtained 197 isolates identified as E. coli by MALDI-TOF MS. A total of 158 representative isolates were whole-genome sequenced, 17 isolates were then reclassified to Escherichia albertii. We observed a higher (86 %; 62/72) occurrence of ESBL/AmpC-producing Escherichia spp. among gulls in 2018 compared to 38 % (17/45) in 2019 (p < 0.00001). The decrease in prevalence was linked to clonal lineage of E. coli ST11893 predominating in 2018 which carried blaCMY-2 and which was not recovered from the gulls in 2019. Oppositely, several Escherichia STs were found in gulls from both years as well as in the water sample including STs commonly recognized as internationally high-risk lineages such as ST10, ST58, ST88, ST117, ST648 or ST744. Phylogenetic analysis of E. coli from EnteroBase from countries where these particular gulls wander revealed that some STs are commonly found in various sources including humans and a portion of them is even closely related (up to 100 SNPs) to our isolates. We demonstrated that the occurrence of AMR in Escherichia can vary greatly in time in synanthropic birds and we detected both, a temporary prevalent lineage and several persistent STs. The close relatedness of isolates from gulls and isolates from EnteroBase highlights the need to further evaluate the risk connected to wandering birds.
Avian pathogenic Escherichia coli (APEC) can cause various extraintestinal infections in poultry, resulting in massive economic losses in poultry industry. In addition, some avian E. coli strains may ...have zoonotic potential, making poultry a possible source of infection for humans. Due to its extreme genetic diversity, this pathotype remains poorly defined. This study aimed to investigate the diversity of colibacillosis-associated E. coli isolates from Central European countries with a focus on the Czech Republic.
Of 95 clinical isolates subjected to preliminary characterization, 32 were selected for whole-genome sequencing. A multi resistant phenotype was detected in a majority of the sequenced strains with the predominant resistance to β-lactams and quinolones being associated with TEM-type beta-lactamase genes and chromosomal gyrA mutations respectively. The phylogenetic analysis confirmed a great diversity of isolates, that were derived from nearly all phylogenetic groups, with predominace of B2, B1 and C phylogroups. Clusters of closely related isolates within ST23 (phylogroup C) and ST429 (phylogroup B2) indicated a possible local spread of these clones. Besides, the ST429 cluster carried bla
genes for AmpC beta-lactamase and isolates of both clusters were generally well-equipped with virulence-associated genes, with considerable differences in distribution of certain virulence-associated genes between phylogenetically distant lineages. Other important and potentially zoonotic APEC STs were detected, incl. ST117, ST354 and ST95, showing several molecular features typical for human ExPEC.
The results support the concept of local spread of virulent APEC clones, as well as of zoonotic potential of specific poultry-associated lineages, and highlight the need to investigate the possible source of these pathogenic strains.
Clostridioides difficile is one of the most important human pathogens. The identification of its possible sources is important for the understanding of C. difficile infection (CDI) epidemiology. A ...total of 16 water samples from wastewater and surface water in South Moravia in the Czech Republic and 82 samples of fish and gulls were collected between May and July 2019. C. difficile isolates were cultured by direct plating and after enrichment on chromogenic media. Susceptibility testing to eight antimicrobials was performed by Etest. C. difficile isolates were characterized by ribotyping, multilocus sequence typing, multilocus tandem repeats analysis, and toxin gene detection. Samples from fish and gulls were C. difficile negative; a total of 15 C. difficile isolates from 8 out of 16 water samples were cultured (6 out of 14 surface water samples yielded 6 isolates, and 2 out of 2 wastewater samples yielded 9 isolates). Direct plating was culture positive in 6 out of 16 samples (12 isolates), and enrichment culture was positive in an additional 2 out of 16 samples (3 isolates). Twelve different ribotyping profiles and 14 sequence types of clades 1, 4, and 5 were identified. Five isolates did not carry genes for toxins, and eight isolates carried genes for toxins A and B; the remaining two isolates (RT078) carried the genes for toxins A, B, and binary. All C. difficile isolates were susceptible to amoxicillin, moxifloxacin, tetracycline, and vancomycin and resistant to ciprofloxacin. A high level of erythromycin resistance (>256 mg/L) was detected in eight isolates. Clindamycin resistance was found in 14 isolates, 6 of which showed a high level of resistance (>256 mg/L) and carried ermB. Surprisingly, one isolate (RT010, ST15) showed resistance to metronidazole (12 mg/L) with the presence of the plasmid pCD-METRO. In conclusion, a diverse spectrum of C. difficile strains was found in wastewater and surface water. A recently discovered plasmid-bound resistance to metronidazole was detected in C. difficile from the surface water sample. IMPORTANCE The combination of direct plating and culture after enrichment was used in order to gain a spectrum of C. difficile ribotypes present in the water samples. Toxigenic C. difficile ribotypes detected in surface water and in wastewater treatment plants overlapped with those derived from patients with CDI and/or animals. Importantly, a recently discovered plasmid-mediated resistance to metronidazole, a drug used for the treatment of CDI, was detected in C. difficile from river water.
Avian colibacillosis is one of the main causes of economic losses in the poultry industry worldwide. Vaccination could help to prevent infection during the laying period on broiler breeder farms. ...Effective vaccination against avian pathogenic
Escherichia coli
(APEC) may be an essential step for protection of poultry flocks depending on the region where they are raised. The aim of this study was to investigate the additive protective effect of an autogenous
E. coli
vaccine in broiler breeders pre-vaccinated with a licensed
E. coli
vaccine (Poulvac®). Our field study was partially blinded and parallel group designed. Group 1 included 24 000 laying hens vaccinated by Poulvac®. Group 2 comprised 12 000 laying hens vaccinated by Poulvac® and additionally, by an autogenous
E. coli
vaccine via intramuscular application before transfer. The effectiveness of vaccination in both groups was evaluated according to the results of gross pathology, bacteriology (isolation and characterization of
E. coli
) and utility indicators. Based on the pathology, the occurrence of
E. coli
polyserositis syndrome (EPS), salpingoperitonitis syndrom (SPS), and haemorrhagic septicaemia was decreased in Group 2 compared to Group 1. The difference in the occurrence of EPS (
P
< 0.001) and SPS (
P
= 0.0342) was significant. The proportion of serotype O78 among
E. coli
isolated from Group 1 and Group 2 was also significant (
P
= 0.0178). The effective and multi-serotype vaccination program in order to expand heterologous protection of laying hens and combination of commercial and autogenous vaccines seems to be a promising preventive management tool.
To investigate a possible Clostridioides difficile reservoir in the Czech Republic, we performed a study in 297 calves from 29 large-scale dairy farms. After enrichment, faecal samples were ...inoculated onto selective agar for C. difficile. From the 297 samples, 44 C. difficile isolates were cultured (prevalence of 14.8%, 10 farms). The Holstein breed and use of digestate were associated with C. difficile colonisation (p ˂ 0.05). C. difficile isolates belonged to the ribotype/sequence type: RT033/ST11 (n = 37), RT126/ST11 (n = 6) and RT046/ST35 (n = 1). A multiple-locus variable-number tandem-repeat analysis revealed four clonal complexes of RT033 isolates and one clonal complex of RT126 isolates. All isolates were sensitive to amoxicillin, metronidazole and vancomycin. Forty isolates were resistant to ciprofloxacin, twenty-one to clindamycin, seven to erythromycin, seven to tetracycline and six to moxifloxacin. Moxifloxacin resistant isolates revealed an amino-acid substitution Thr82Ile in the GyrA. In conclusion, the calves of Holstein breed from farms using digestate as a product of bio-gas plants are more likely to be colonised by clonally-related C. difficile of ST 11 represented by ribotypes 033 and 126. The identified resistance to moxifloxacin with a Thr82Ile substitution in the GyrA highlights the need for further monitoring by the “One health approach”.
Antimicrobial resistance genes can be found in all ecosystems, including those where antibiotic selective pressure has never been exerted. We investigated resistance genes in a collection of faecal ...samples of wildlife (non-human primates, mice), people and domestic animals (dogs, cats) in Côte d'Ivoire; in the chimpanzee research area of Taï National Park (TNP) and adjacent villages. Single bacteria isolates were collected from antibiotic-containing agar plates and subjected to molecular analysis to detect Enterobacteriaceae isolates with plasmid-mediated genes of extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated quinolone resistance (PMQR). While the prevalence of ESBL-producing E. coli in the villages was 27% in people (n = 77) and 32% in dogs (n = 38), no ESBL-producer was found in wildlife of TNP (n = 75). PMQR genes, mainly represented by qnrS1, were also present in human- and dog-originating isolates from the villages (36% and 42% in people and dogs, respectively), but no qnrS has been found in the park. In TNP, different variants of qnrB were detected in Citrobacter freundii isolates originating non-human primates and mice. In conclusion, ESBL and PMQR genes frequently found in humans and domestic animals in the villages were rather exceptional in wildlife living in the protected area. Although people enter the park, the strict biosecurity levels they are obliged to follow probably impede transmission of bacteria between them and wildlife.
The aim of the present study was to monitor the presence of extended-spectrum beta-lactamase (ESBL) producing
E. coli
on farm A with the history of previous use of ceftiofur in suckling pigletsand to ...analyse the risk factors of selection and dissemination of ESBL producers in the production herd. In the year of 2014, a total of 411 samples (rectal swabs or faeces)from pigs of various age categories (sows, gilts and suckling piglets) were collected. The sampling was performed more than 24 months after the ban of ceftiofur on the farm.The sows and gilts were sampled repeatedly before and after farrowing. All collected samples were directly cultivated on MacConkey agar (MCA) containing cefotaxime (2 mg/l) and obtained sub-cultures were tested for ESBL production by double disc synergy test. According to our results, all gilts were negative for ESBL-producing
E. coli
in the introduction period, however, the excretion of ESBL-producing
E. coli
was observed before and after delivery. Most of the new-born piglets from positive sows and gilts shed ESBL-producing
E. coli
early after birth. All tested ESBL-producing isolates were resistant to multiple antimicrobials, suggesting that antibiotics from other groups used for therapy co-select for ESBL producers in pigs on the studied farm. Intestinal colonization of lactating sows and their offspring as well as survival of ESBL-producing
E. coli
in the farm environment should be recognised as important risk factors of circulation and long-time persistence of ESBL producers in the herd.
The aim of this study was to help improve the management of assisted reproduction of donkeys and to extend the existing information on the fertility of donkeys by qualitative and quantitative ...assessment of bacterial contamination of the jennies’ genital tract in connection with the perineum formation and oestrous cycle phases. Ten female donkeys were included in the study and were repeatedly rectally palpated and sonographically examined during the oestrous cycle. Samples were taken from the fossa clitoridis and around the cervix for microbiological examination. Each jenny was sampled twice, always at different stages of the cycle, both in the oestrus and in the dioestrus. In addition, samples from the endometrium were taken in oestrus for both microbiological and cytological examination. After collection, the samples were examined in a microbiological laboratory. There were 62 different species of microorganisms found belonging to 19 different bacterial genera. The presence of agents of mares’ infectious metritis
Taylorella equigenitalis
was not found in any of the samples. The outcome of our study is an initial mapping of the microbial colonization of the endometrium and the genital tract in jennies with correct formation of the perineal region. The hypothesis that the level of bacterial contamination was related to the oestrous cycle was not confirmed. The practical significance of the study lies mainly in the description of the composition, amount and changes of microbiota in healthy individuals during their oestrous cycle, which allows the evaluation of the risk of developing infection during the active oestrous cycle in connection with mating.
The aims of this study were to characterize the choanal and cloacal aerobic bacterial flora in healthy captive green iguanas and to compare it with the bacterial flora of the biofilm present in the ...water container of each terrarium. Samples were collected from the choana and the cloaca of 20 healthy captive adult green iguanas and from the biofilm of 15 water containers. The final identification of aerobic bacteria was performed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.
Salmonella
positive samples were serotyped. The most common strains observed at each test location were from 1) choanae:
Staphylococcus
spp.,
Enterobacter cloacae
and
Comamonas testosteroni
; 2) cloacae:
Citrobacter
spp.,
Salmonella
spp. and
Corynebacterium
spp.; and 3) biofilms:
Pseudomonas
spp.,
Salmonella
spp. and
Acidovorax
spp. We showed that apart from
Salmonella
spp., the choanal and cloacal bacterial flora differed from the microorganisms present in the biofilm of the animal’s water container. These data revealed that healthy captive adult green iguanas harbored several aerobic bacterial strains that in immunosuppressed reptiles may act as opportunistic pathogens. Also, several of the aerobic bacteria identified in samples are potential zoonotic agents. Characterization of the normal background flora in captive reptiles and their environment can contribute to an understanding of the spread of bacterial contamination and the risk of potential zoonotic diseases for people in contact with these animals.
The objective of this study was to investigate the silver gull as an indicator of environmental contamination by salmonellae and carbapenemase-producing Enterobacteriaceae (CPE) in south-east ...Australia.
A total of 504 cloacal samples were collected from gull chicks at three nesting colonies in New South Wales, Australia White Bay (n = 144), Five Islands (n = 200) and Montague Island (n = 160) and were examined for salmonellae and CPE. Isolates were tested for carbapenemase genes and susceptibility to 14 antibiotics. Clonality was determined by PFGE and MLST. Genetic context and conjugative transfer of the carbapenemase gene were determined.
A total of 120 CPE of 10 species, mainly Escherichia coli (n = 85), carrying the gene blaIMP-4, blaIMP-38 or blaIMP-26 were obtained from 80 (40%) gulls from Five Islands. Thirty percent of birds from this colony were colonized by salmonellae. Most isolates contained the gene within a class 1 integron showing a blaIMP-4-qacG-aacA4-catB3 array. The blaIMP gene was carried by conjugative plasmids of variable sizes (80-400 kb) and diverse replicons, including HI2-N (n = 30), HI2 (11), A/C (17), A/C-Y (2), L/M (5), I1 (1) and non-typeable (6). Despite the overall high genetic variability, common clones and plasmid types were shared by different birds and bacterial isolates, respectively.
Our data demonstrate a large-scale transmission of carbapenemase-producing bacteria into wildlife, likely as a result of the feeding habits of the birds at a local waste depot. The isolates from gulls showed significant similarities with clinical isolates from Australia, suggesting the human origin of the isolates. The sources of CPE for gulls on Five Islands should be explored and proper measures applied to stop the transmission into the environment.
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