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1.
BMC Infect Dis ; 20(1): 278, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32293315

RESUMO

BACKGROUND: Understanding potential risks of multi-drug resistant (MDR) pathogens from the booming poultry sector is a crucial public health concern. Campylobacter spp. are among the most important zoonotic pathogens associated with MDR infections in poultry and human. This study systematically examined potential risks and associated socio-environmental factors of MDR Campylobacter spp. in poultry farms and live bird markets (LBMs) of Bangladesh. METHODS: Microbial culture and PCR-based methods were applied to examine the occurrence and MDR patterns of Campylobacter spp. in potential sources (n = 224) at 7 hatcheries, 9 broiler farms and 4 LBMs in three sub-districts. Antimicrobial residues in broiler meat and liver samples (n = 50) were detected by advanced chromatographic techniques. A questionnaire based cross-sectional survey was conducted on socio-environmental factors. RESULTS: Overall, 32% (71/ 224) samples were found contaminated with Campylobacter spp. In poultry farms, Campylobacter spp. was primarily found in cloacal swab (21/49, 43%), followed by drinking water (8/24, 33%), and meat (8/28, 29%) samples of broilers. Remarkably, at LBMs, Campylobacter spp. was detected in higher prevalence (p < 0.05) in broiler meat (14/26, 54%), which could be related (p < 0.01) to bacterial contamination of drinking water (11/21, 52%) and floor (9/21, 43%). Campylobacter isolates, one from each of 71 positive samples, were differentiated into Campylobacter jejuni (66%) and Campylobacter coli (34%). Alarmingly, 49 and 42% strains of C. jejuni and C. coli, respectively, were observed as MDR, i.e., resistant to three or more antimicrobials, including, tetracycline, amoxicillin, streptomycin, fluoroquinolones, and macrolides. Residual antimicrobials (oxytetracycline, ciprofloxacin and enrofloxacin) were detected in majority of broiler liver (79%) and meat (62%) samples, among which 33 and 19%, respectively, had concentration above acceptable limit. Inadequate personal and environmental hygiene, unscrupulously use of antimicrobials, improper waste disposal, and lack of health surveillance were distinguishable risk factors, with local diversity and compound influences on MDR pathogens. CONCLUSION: Potential contamination sources and anthropogenic factors associated with the alarming occurrence of MDR Campylobacter, noted in this study, would aid in developing interventions to minimize the increasing risks of poultry-associated MDR pathogens under 'One Health' banner that includes poultry, human and environment perspectives.


Assuntos
Antibacterianos/análise , Infecções por Campylobacter/epidemiologia , Campylobacter coli/efeitos dos fármacos , Campylobacter jejuni/efeitos dos fármacos , Farmacorresistência Bacteriana Múltipla/genética , Fazendas , Doenças das Aves Domésticas/epidemiologia , Aves Domésticas/microbiologia , Animais , Antibacterianos/uso terapêutico , Bangladesh/epidemiologia , Infecções por Campylobacter/tratamento farmacológico , Infecções por Campylobacter/microbiologia , Campylobacter coli/genética , Campylobacter coli/isolamento & purificação , Campylobacter jejuni/genética , Campylobacter jejuni/isolamento & purificação , Galinhas/microbiologia , Estudos Transversais , Humanos , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Prevalência , Fatores de Risco
2.
Int J Food Microbiol ; 322: 108564, 2020 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-32163798

RESUMO

Campylobacter is one of the most important causative agents of foodborne illnesses worldwide. The poultry reservoir is the main source of Campylobacter. Within the broiler production chain, campylobacters can only multiply in the chicken's intestinal tract. Intervention at farm level to reduce Campylobacter is thus preferred, but despite extensive study, no highly effective solutions have been found to combat Campylobacter at farm level. Slaughterhouses are experiencing great pressure to deliver carcasses with low Campylobacter contamination even when they receive and slaughter Campylobacter colonized flocks. Since 2018, a process hygiene criterion (EU 2017/1495) with the critical limit of <1000 cfu/g neck skin has been implemented in EU Member States based on the calculation done at the time of the study that human campylobacteriosis cases could be halved if all carcasses would comply with a criterion of <1000 cfu/g neck skin. This review covers Campylobacter contamination of broiler carcasses from transport through the different slaughter steps. Possible intervention methods during slaughter are discussed with a focus on the European situation, where chemicals are not allowed to disinfect carcasses.


Assuntos
Matadouros/normas , Campylobacter/isolamento & purificação , Manipulação de Alimentos/normas , Microbiologia de Alimentos/normas , Aves Domésticas/microbiologia , Animais , Galinhas , Europa (Continente)/epidemiologia , Contaminação de Alimentos/análise , Contaminação de Alimentos/prevenção & controle , Manipulação de Alimentos/métodos , Microbiologia de Alimentos/métodos , Higiene/normas
3.
Int J Food Microbiol ; 322: 108572, 2020 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-32169770

RESUMO

Global dissemination of non-typhoidal Salmonella producing extended-spectrum ß-lactamase (ESBL) is a public-health concern. Recently, the prevalence of Salmonella spp. resistant to third-generation cephalosporins has been increasing in food-producing animals in Korea. In this study, we investigated resistance mechanisms and molecular characteristics of S. Virchow isolates resistant to extended-spectrum cephalosporins (ESCs). We obtained 265 S. Virchow isolates from fecal and carcasses samples of cattle (n = 2), pigs (n = 7), and chickens (n = 256) during 2010-2017, and observed high ESC-resistance (63.8%, 169/265); most of the resistant isolates (96.4%) were obtained from chickens. ESC-resistant S. Virchow isolates (n = 169) showed significantly higher resistance rates to other antimicrobials (especially aminoglycosides and tetracycline, p-value <0.0001), as well as prevalence of multidrug resistance, than did ESC-susceptible S. Virchow isolates (n = 96). All ESC-resistant S. Virchow produced CTX-M-15-type ESBL (n = 147) and/or CMY-2-type AmpC ß-lactamase (n = 23). ESC-resistant S. Virchow represented seven pulsotypes, predominantly composed of type II (58.6%) and III (26.0%), detected in 69 farms in 10 provinces, and 33 farms in 7 provinces, respectively. Genes encoding ESC-resistance were horizontally transferred by conjugation to recipient E. coli J53; this was demonstrated in 28.8% (42/146) of blaCTX-M-15-positive isolates and in 50.0% (11/22) of blaCMY-2-positive isolates. All conjugative plasmids carrying blaCTX-M-15 and blaCMY-2 genes belonged to ST2-IncHI2 and ST12/CC12-IncI1, respectively. Genetic features of transferred bla genes were involved with ISEcp1 in both blaCTX-M-15 and blaCMY-2; ISEcp1 plays a critical role in the efficient capture, expression, and mobilization of bla genes. In addition to blaCTX-M-15 genes, resistance markers to aminoglycosides and/or tetracycline were co-transferred to recipient E. coli J53. Our results show a high prevalence of ESBL-producing S. Virchow in chickens and chicken carcasses. Specific blaCTX-M-15 and blaCMY-2-carrying S. Virchow clones and plasmids were predominant in food-producing animals nationwide. Restriction of antimicrobial use and proper biosecurity practices at the farm level should be urgently implemented in the poultry industry.


Assuntos
Microbiologia de Alimentos , Salmonella enterica/enzimologia , Salmonella enterica/isolamento & purificação , beta-Lactamases/genética , Animais , Antibacterianos/farmacologia , Proteínas de Bactérias , Bovinos/microbiologia , Cefalosporinas/farmacologia , Galinhas/microbiologia , Farmacorresistência Bacteriana/efeitos dos fármacos , Farmacorresistência Bacteriana/genética , Plasmídeos/genética , Aves Domésticas/microbiologia , República da Coreia , Salmonella enterica/efeitos dos fármacos , Salmonella enterica/genética , Sorogrupo , Suínos/microbiologia , beta-Lactamases/metabolismo
4.
BMC Infect Dis ; 20(1): 192, 2020 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-32131753

RESUMO

BACKGROUND: Human psittacosis, caused by Chlamydia (C.) psittaci, is likely underdiagnosed and underreported, since tests for C. psittaci are often not included in routine microbiological diagnostics. Source tracing traditionally focuses on psittacine pet birds, but recently other animal species have been gaining more attention as possible sources for human psittacosis. This review aims to provide an overview of all suspected animal sources of human psittacosis cases reported in the international literature. In addition, for each animal species the strength of evidence for zoonotic transmission was estimated. METHODS: A systematic literature search was conducted using four databases (Pubmed, Embase, Scopus and Proquest). Articles were included when there was mention of at least one human case of psittacosis and a possible animal source. Investigators independently extracted data from the included articles and estimated strength of evidence for zoonotic transmission, based on a self-developed scoring system taking into account number of human cases, epidemiological evidence and laboratory test results in human, animals, and the environment. RESULTS: Eighty articles were included, which provided information on 136 different situations of possible zoonotic transmission. The maximum score for zoonotic transmission was highest for turkeys, followed by ducks, owls, and the category 'other poultry'. Articles reporting about zoonotic transmission from unspecified birds, psittaciformes and columbiformes provided a relatively low strength of evidence. A genotypical match between human and animal samples was reported twenty-eight times, including transmission from chickens, turkeys, guinea fowl, peafowl, pigeons, ducks, geese, songbirds, parrot-like birds and owls. CONCLUSIONS: Strong evidence exists for zoonotic transmission from turkeys, chickens and ducks, in addition to the more traditionally reported parrot-like animal sources. Based on our scoring system, the evidence was generally stronger for poultry than for parrot-like birds. Psittaciformes should not be disregarded as an important source of human psittacosis, still clinicians and public health officials should include poultry and birds species other than parrots in medical history and source tracing.


Assuntos
Chlamydophila psittaci/genética , Chlamydophila psittaci/imunologia , Doenças das Aves Domésticas/transmissão , Aves Domésticas/microbiologia , Psitacose/diagnóstico , Zoonoses/transmissão , Animais , Columbidae/microbiologia , Genótipo , Humanos , Psitacose/microbiologia , Saúde Pública , Administração em Saúde Pública , Aves Canoras/microbiologia , Estrigiformes/microbiologia
5.
J Environ Sci Health B ; 55(2): 155-165, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31985354

RESUMO

Salmonella and Campylobacter are significant issues for poultry processors because of increasing regulatory standards as well as public health concerns. The goal of this study is to report the effects of two different pre-chiller systems that utilize different temperatures and water recirculation systems on whole bird carcass rinsates. Both pre-chiller tanks were contained within a single poultry processing facility and operated at different temperatures and water systems. The incidence of Campylobacter spp. and Salmonella spp., as well as the aerobic plate counts on whole bird carcass rinses are reported in this study from each pre-chiller system. The results from this study reveal that there are significant differences in how microbial populations and pathogens change over time in each pre-chiller system. Furthermore, we identify that these patterns are different per system. Such data are impactful as it indicates that measuring carcasses within a plant must consider both temperature and water recirculation as it may prevent comparability of different lines within a single processing facility.


Assuntos
Campylobacter , Microbiologia de Alimentos , Indústria de Processamento de Alimentos/métodos , Aves Domésticas/microbiologia , Salmonella , Animais , Carga Bacteriana , Indústria de Processamento de Alimentos/instrumentação , Temperatura , Água
6.
Int J Food Microbiol ; 314: 108391, 2020 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-31689613

RESUMO

Poultry production is the fastest growing meat sector worldwide. In the last five years, growing concerns have been expressed by international health agencies and consumers about the transmission of antibiotic-resistant bacteria from poultry meat to human. Consequently, poultry producers have adopted alternative production systems based on reduced antibiotic usage, including organic and antibiotic-free (AF) production. However, the effect of these production systems on the antibiotic resistance of the gut flora in slaughtered poultry has been poorly investigated. We hypothesized that organic and AF production systems reduce the risk of antibiotic resistance in the commensal Escherichia coli of broilers at slaughter compared with conventional production. Cecal content from broilers raised in conventional (292), AF (291), or organic (272) flocks (855 broilers in total) belonging to the same company was sampled. E. coli loads [colony-forming units (CFU/g)] and numbers of E. coli resistant to nalidixic acid (E. colinal) were determined for each sample. Antibiotic susceptibility of one isolate per sample was evaluated using the disc diffusion method; colistin resistance was determined by using the broth microdilution method. The differences in bacterial loads from the three production types were evaluated using one-way ANOVA. Differences in the proportion of resistant isolates in the three production lines were evaluated using Pearson's χ2 or Fisher's test. The strength of the association was evaluated by using odds ratio (OR), with the conventional production type as a reference (OR = 1). Overall, the analysis revealed a high level of resistance (50% or higher) to ampicillin, cefazolin, sulfonamides, nalidixic acid, and tetracycline, independently of the production type. High proportion of ciprofloxacin resistance (52%) was observed, with 4.5% isolates resistant to cefotaxime and 1.8% resistant to colistin. The average loads (log CFU/g cecal content) of E. colinal were determined as 6.84 for AF, 6.38 for organic type, and 7.27 for conventional type. The difference was significant (p < 0.00001). Interestingly, broilers from AF flocks had higher E. colinal loads than broilers from organic flocks. This trend (conventional > AF > organic) was confirmed by qualitative data. However, the magnitude of the effect, measured as a reduced risk of resistance, varied broadly for the antibiotics tested. These findings suggest that poultry production systems alternative to the conventional broiler production are associated with reduced frequency of antibiotic-resistant E. coli among the commensal gut flora, posing a lower risk to the environment and the consumer.


Assuntos
Agricultura/métodos , Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Infecções por Escherichia coli/veterinária , Escherichia coli/efeitos dos fármacos , Escherichia coli/isolamento & purificação , Aves Domésticas/microbiologia , Animais , Antibacterianos/administração & dosagem , Ceco/microbiologia , Galinhas/microbiologia , Escherichia coli/crescimento & desenvolvimento , Infecções por Escherichia coli/microbiologia , Testes de Sensibilidade Microbiana
7.
Int J Food Microbiol ; 318: 108478, 2020 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-31855787

RESUMO

Avian industries have been reported as an important contributor in the worldwide spread of antibiotic resistance owing to some particular practices especially the overuse of antibiotics. Thus in this study, we aimed to characterize extended-spectrum-beta-lactamase (ESBL) and acquired-AmpC-beta-lactamase (aAmpC)-producing Escherichia coli isolates from chicken faeces and raw meat in Tunisia. During the year 2018, 286 faecal chicken swabs and 47 raw chicken meat samples were collected and processed to recover cefotaxime-resistant E. coli. Antimicrobial susceptibility was performed by disk-diffusion and/or broth-microdilution. blaTEM, blaSHV, blaCTX-M, and blaCMY genes were investigated by PCR/sequencing. Genes encoding resistance to colistin (mcr-1 to mcr-8), tetracycline (tetA/tetB), sulfonamide (sul1/sul3), and chloramphenicol (cmlA), were analysed by PCR. Class 1 integrons were investigated by PCR/sequencing. Phylogenetic groups of all isolates were determined. PFGE and MLST were performed for representative isolates. PCR-based replicon typing was performed in mcr1-harbouring isolates. Cefotaxime-resistant E. coli was detected in 22.4% (64/286) and 63.8% (30/47) of faeces and meat samples, respectively. Ninety isolates were ESBL-producers and harboured the genes: blaCTX-M-1 +/- blaTEM-1 (n = 65), blaCTX-M-55 +/- blaTEM-1 (n = 21), blaCTX-M-14 (n = 1), and blaSHV-12 (n = 3). The blaCMY-2 gene was detected in four ESBL-negative isolates. Isolates belonged to phylogroups D (50%), A (36.2%), B1 (9.6%), and B2 (4.3%). Fifty-four were colistin-resistant and 52 carried the mcr-1 gene. The tetA, sul1/sul3 and cmlA genes were detected among resistant isolates and 76 harboured class 1 integrons. MLST analysis revealed 13 sequence types (STs). The isolates were classified into 28 PFGE types. The IncP, IncFIB, and IncI1 replicons were detected among mcr-1-positive strains. We report a high frequency of ESBL-producers and colistin-resistant E. coli in chicken and derived food and the detection for the first time of blaCTX-M-55 in poultry in Tunisia.


Assuntos
Farmacorresistência Bacteriana/genética , Proteínas de Escherichia coli/genética , Escherichia coli/genética , Aves Domésticas/microbiologia , beta-Lactamases/genética , Animais , Antibacterianos/farmacologia , Galinhas , Farmacorresistência Bacteriana/efeitos dos fármacos , Escherichia coli/classificação , Escherichia coli/efeitos dos fármacos , Escherichia coli/isolamento & purificação , Proteínas de Escherichia coli/metabolismo , Fezes/microbiologia , Filogenia , Prevalência , Tunísia , beta-Lactamases/metabolismo
8.
Food Chem ; 309: 125690, 2020 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-31711808

RESUMO

In this work a new fluorescence immunosensor with use of graphene oxide and graphene quantum dot for detection Campylobacter jejuni whole cell in food samples was designed. This biosensor was designed based on interaction of poly clonal antibody conjugated with graphene quantum dot with surface protein in Campylobacter jejuni cell membrane. Specific binding of graphene quantum dot with Campylobacter jejuni membrane leads to generate a distance among graphene dot and graphene oxide and fluorescence is ON. In lack of Campylobacter jejuni or in existence of other bacterial cells, distance between of graphene dot and graphene oxide is very low and graphene quantum dot fluorescence emission was OFF. Experiment revealed that step by step increase in bacterial target cells caused to gradually increased fluorescence emission and this process was linear. Limit of detection for this bacterial sensor was 10 CFU/ml and ability of this FRET immunosensor for Campylobacter jejuni sensing in comparison with other bacterial cells was significant. Also, this method for monitoring Campylobacter jejuni in poultry liver was applied and results revealed that this immunosensor could be used for analysis bacterial cell in food samples.


Assuntos
Técnicas Biossensoriais/métodos , Campylobacter jejuni/isolamento & purificação , Grafite/química , Pontos Quânticos/química , Animais , Anticorpos Antibacterianos/química , Anticorpos Antibacterianos/imunologia , Proteínas da Membrana Bacteriana Externa/imunologia , Campylobacter jejuni/metabolismo , Transferência Ressonante de Energia de Fluorescência , Microbiologia de Alimentos , Limite de Detecção , Fígado/microbiologia , Aves Domésticas/microbiologia
9.
Int J Food Microbiol ; 312: 108358, 2020 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-31655356

RESUMO

Meat products are commonly regarded as one of the main sources of human listeriosis caused by Listeria monocytogenes. The objective of this study was to estimate the prevalence of L. monocytogenes in a range of meat products from 24 different Chinese regions by using meta-analysis of literature data and a novel sensitivity analysis approach. A total of 112 publications from five databases, published between 1 January 2007 and 31 December 2017, were systematically selected for relevance and covered meat products sampled between 2000 and 2016. Estimated by the random-effects model, the pooled prevalence of L. monocytogenes was 8.5% (95% CI: 7.1%-10.3%) in raw meats and 3.2% (95% CI: 2.7%-3.9%) in ready-to-eat (RTE) meats. The prevalence differed from high to low among raw meats including prefabricated raw meats 12.6% (95% CI: 6.9%-21.7%), fresh pork 11.4% (95% CI: 8.6%-14.9%), fresh beef 9.1% (95% CI: 6.3%-13.0%), fresh poultry 7.2% (95% CI:4.9%-10.4%), frozen raw meats 7.2% (95% CI: 5.7%-9.0%), and fresh mutton 5.4% (95% CI: 2.5%-11.0%). A higher L. monocytogenes prevalence level was shown in the meat products from central and northeastern China provincial regions. The entropy-based sensitivity analysis utilized in the meta-analysis indicated that the sampling period and location were two critical factors influencing the prevalence level of L. monocytogenes in meat products. A better understanding of differences in prevalence levels per geographic region and between meat product sources may allow the competent authorities, industry, and other relevant stakeholders to tailor their interventions to control the occurrence of L. monocytogenes in meat products effectively.


Assuntos
Contaminação de Alimentos/análise , Microbiologia de Alimentos , Listeria monocytogenes/isolamento & purificação , Produtos da Carne/microbiologia , Aves Domésticas/microbiologia , Animais , Bovinos , China , Humanos , Listeriose/epidemiologia , Prevalência , Ovinos , Suínos
10.
Comp Immunol Microbiol Infect Dis ; 66: 101345, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31476607

RESUMO

Campylobacter jejuni is a major cause of infectious diarrhea in humans. The bacterium can be transmitted through contaminated poultry meat and waste water. We report the presence of C. jejuni from potential transmission sources including egg shells, poultry waste, waste water and migratory bird droppings with a prevalence rate of 78%, 66%, 86% and 70% respectively. Antibiotic resistance profile showed high number of isolates resistant to multiple antibiotics including 4th generation cephalosporins. C. jejuni isolates were further screened for presence of T6SS, an important virulence factor. None of the C. jejuni isolates from migratory birds carried a T6SS, whereas highest prevalence of T6SS isolates was observed in waste water samples, followed by poultry waste and egg shells. To determine virulence potential of the isolates, hemolytic activity of isolates was compared. Although variation in hemolytic potential between isolates from different sources was noted, higher hemolytic activity was observed for isolates possessing hcp, a T6SS gene. Furthermore, presence of T6SS affords the bacterium some survival advantage when compared to T6SS competent Helicobacter pullorum which occupies the same niche. Taken together our findings indicate that C. jejuni with T6SS have a fitness advantage increasing their isolation frequency from waste water and poultry waste.


Assuntos
Antibacterianos/farmacologia , Infecções por Campylobacter/veterinária , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/patogenicidade , Sistemas de Secreção Tipo VI/genética , Migração Animal , Animais , Aves/microbiologia , Infecções por Campylobacter/transmissão , Campylobacter jejuni/genética , DNA Bacteriano/genética , Casca de Ovo/microbiologia , Aves Domésticas/microbiologia , Virulência , Águas Residuárias/microbiologia
11.
Food Microbiol ; 84: 103237, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31421774

RESUMO

Between 1991 and 2014 the per capita notification rate of salmonellosis in Australia increased from 31.9 to 69.7 cases per 100,000 people. Salmonella Typhimurium accounted for nearly half the human cases until the end of 2014. In this study, we used cluster analysis tools to compare S. Typhimurium isolates from a chicken-meat study with those reported to the National Enteric Pathogen Surveillance System (NEPSS) from the coincident human and non-human populations. There was limited phage type diversity within all populations and a lack of specificity of MLVA profiling within phage types. The chicken-meat study isolates were not significantly clustered with the human cases and at least 7 non-human sources, based on typing profiles (PT/MLVA combination), could be implicated as a source of human cases during the same period. In the absence of a strong surveillance system representative of all putative sources, MLVA and phage typing alone or in combination are insufficient to identify the source of human cases.


Assuntos
Tipagem de Bacteriófagos , Surtos de Doenças , Fagos de Salmonella/classificação , Salmonella typhimurium/classificação , Animais , Técnicas de Tipagem Bacteriana , Bovinos , Galinhas , Análise por Conglomerados , Humanos , Repetições Minissatélites , Aves Domésticas/microbiologia , Intoxicação Alimentar por Salmonella/diagnóstico , Intoxicação Alimentar por Salmonella/microbiologia , Infecções por Salmonella/diagnóstico , Infecções por Salmonella/microbiologia , Salmonella typhimurium/virologia
12.
Ecotoxicol Environ Saf ; 183: 109514, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31394374

RESUMO

This study was conducted to determine the prevalence of integrons and associated gene cassettes in Escherichia coli (E. coli) isolates from waterfowls in Hainan, China. The antimicrobial resistance profile of the isolates was examined by using disc diffusion test. In addition, PCR, RFLP, plasmid replicon typing and DNA sequencing analyses were used for the characterization of integrase genes (class 1, 2 and 3) and associated gene cassettes. Approximatively, 90% of the isolates were positive for the integrase genes by PCR. Specifically, class 1 and class 2 integrons were found in 252 (81%) and 7 (2.3%) strains, respectively. While 21 (6.7%) isolates were positive for both class 1 and class 2 integrons. However, none of the isolate was positive for the class 3 integrons. In addition, 5 various cassette arrays, dfrA1-orfC, aadA2, aadA1, dfrA1-aadA1, and dfrA1-orfC- aadA1, were found within the variable regions (VRs) of class 1 integron isolates. While only single cassette array, dfrA1-sat2- aadA1, was identified within VRs of class 2 integron isolates. We identified incF plasmid as the most common plasmid type, which was detected in 81 of 243 VRs containing isolates. This study is the first report showing the baseline characteristics of integrons in E. coli isolates from waterfowls in Hainan, China. Our results provide evidence of the waterfowl birds as a reservoir of class 1 and class 2 integrons carrying antibiotic resistance gene cassettes. Therefore, strict preventive measures should be taken to avoid the spread of mobile genetic resistance elements in waterfowls in China.


Assuntos
Anseriformes/microbiologia , DNA Bacteriano/genética , Farmacorresistência Bacteriana , Escherichia coli/isolamento & purificação , Integrons , Aves Domésticas/microbiologia , Animais , Antibacterianos/farmacologia , China , Farmacorresistência Bacteriana/efeitos dos fármacos , Farmacorresistência Bacteriana/genética , Escherichia coli/genética , Infecções por Escherichia coli/microbiologia , Humanos , Integrons/genética
13.
PLoS One ; 14(7): e0219795, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31329622

RESUMO

Multidrug-resistant Salmonella enterica serovar Heidelberg isolates are frequently recovered in the Netherlands from poultry meat imported from South America. Our aim was to retrospectively assess the characteristics of the antimicrobial determinants, gene content and the clonal relatedness of 122 unique S. Heidelberg isolates from chicken meat from Brazil (n = 119) and Argentina (n = 3) that were imported between 2010 and 2015. These isolates were subjected to antimicrobial susceptibility testing, PCR and Illumina HiSeq2500 whole genome sequencing. Draft genomes were assembled to assess the gene content, and the phylogenetic relationships between isolates were determined using single nucleotide polymorphisms. Ciprofloxacin-resistance was identified in 98.4% of the isolates and 83.7% isolates showed resistance to the extended-spectrum cephalosporins cefotaxime and ceftazidime (83.6% and 82.8% respectively). Of the latter, 97.1% exhibited an AmpC phenotype and contained blaCMY-2, whereas the remaining three isolates contained an extended spectrum beta-lactamase. Of the 99 extended-spectrum cephalosporins-resistant isolates harboring CMY-2 plasmids, 56.6% contained the incompatibility group I1 replicon. Phylogenetic cluster analysis showed that all isolates from Brazil clustered together, with 49% occurring in clusters larger than 5 isolates that revealed intra-cluster similarities based on geographical location and/or resistance profiles. The remaining isolates were classified in smaller clusters or as singletons, highlighting the large diversity of S. Heidelberg in the poultry chain in Brazil that was revealed by this study. Considering the potential public health risk associated with multidrug-resistant S. Heidelberg in imported poultry, collaborative whole genome sequencing-based surveillance is needed to monitor the spread, pathogenic properties and epidemiological distribution of these isolates.


Assuntos
Farmacorresistência Bacteriana Múltipla/genética , Carne/microbiologia , Aves Domésticas/microbiologia , Salmonella enterica/genética , Salmonella enterica/isolamento & purificação , Sorogrupo , Sequenciamento Completo do Genoma , Animais , Antibacterianos/farmacologia , Análise por Conglomerados , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Genes Bacterianos , Tipagem de Sequências Multilocus , Países Baixos , Polimorfismo de Nucleotídeo Único/genética , Salmonella enterica/efeitos dos fármacos
14.
Comp Immunol Microbiol Infect Dis ; 65: 194-200, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31300113

RESUMO

The aims of this study were to regain new epidemiology information about frequency, drug resistance rates, and typing of Campylobacter jejuni (C. jejuni) isolates, obtained from some poultry and cattle farms, slaughterhouses, and people with diarrhea. In this regard, Minimal Inhibitory Concentration (MIC) of several antibiotics and the associated antibiotic resistance genes, including tetO, tetA, cmeB, and blaOXA-61 were evaluated. The isolates were also typed, using the Fla-RFLP method. Generally, between 233 food animal samples, 80 (34.33%) C. jejuni were isolated. Moreover, 20 out of 74 (27%) human specimens suspected to infectious diarrhea were C. jejuni positive. High frequencies of resistance to tetracycline (100%), ciprofloxacin (95%), and nalidixic acid (86%), and low frequencies of resistance to florfenicol (0%), erythromycin (5%), and gentamicin (8%) were observed. Furthermore, in the tetracycline-resistant isolates, the existences of tetO, tetA, and cmeB were 86%, 23%, and 48%, respectively. There was a significant correlation between the cluster types obtained from Fla-RFLP method and antibiotic resistance pattern. The results suggested that the genomic link between Campylobacter spp. should be always evaluated in each country to provide an insight about the Campylobacter spp., spread in the region, in order to implement the health-controlling programs efficiently.


Assuntos
Infecções por Campylobacter/epidemiologia , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/genética , Diarreia/microbiologia , Farmacorresistência Bacteriana Múltipla/genética , Aves Domésticas/microbiologia , Matadouros , Animais , Antibacterianos/farmacologia , Campylobacter jejuni/classificação , Bovinos/microbiologia , Diarreia/epidemiologia , Humanos , Irã (Geográfico)/epidemiologia , Testes de Sensibilidade Microbiana , Tipagem Molecular , Polimorfismo de Fragmento de Restrição , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/microbiologia , Carne Vermelha/microbiologia
15.
Anal Bioanal Chem ; 411(23): 6067-6080, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31273413

RESUMO

Rapid detection of trace Salmonella is urgently needed to ensure food safety. We present an innovative pretreatment strategy, based on a two-step enrichment culture and immunomagnetic separation, combined with a chemiluminescence microparticle immunoassay to detect at least one proliferative Salmonella cell in 25 mL (25 g) food. The capture performance of immunomagnetic beads (IMBs) of sizes for Salmonella was investigated, and the IMBs of size 2.8 µm showed a high capture efficiency of 60.7% in 25 mL milk and 74.5% in 25 mL chicken culture filtrate, which ensured the successful capture of trace Salmonella after 2.5 h in situ enrichment even from only one Salmonella cell. The separated Salmonella cells, reaching an amount of 103 colony-forming units (CFU) by a secondary enrichment for 3 h, were detected by a horseradish peroxidase chemiluminescence reaction with 4-(1-imidazolyl)phenol as an enhancer, which evidenced a linear response for Salmonella concentrations ranging from 2.3 × 102 to 7.8 × 104 CFU/mL. The entire detection process was completed within 8 h, with a very low detection limit of 1 CFU/25 mL (25 g), which was verified by colony counting, and a small degree of interference of 0.17-1.06%. Trace Salmonella from five different serovars in milk and chicken was successfully detected without false negative or false positive results. Furthermore, this study provides a basis to develop a fully automated instrument based on IMBs that includes all steps from sample preparation to chemiluminescence microparticle immunoassay for high-throughput screening of foodborne pathogens. Graphical abstract.


Assuntos
Contaminação de Alimentos/análise , Medições Luminescentes/métodos , Leite/microbiologia , Aves Domésticas/microbiologia , Salmonella/isolamento & purificação , Animais , Galinhas/microbiologia , Contaminação de Alimentos/economia , Inocuidade dos Alimentos/métodos , Imunoensaio/economia , Imunoensaio/métodos , Separação Imunomagnética/economia , Separação Imunomagnética/métodos , Limite de Detecção , Medições Luminescentes/economia , Infecções por Salmonella/microbiologia , Fatores de Tempo
16.
PLoS One ; 14(7): e0220054, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31339918

RESUMO

In this study, we compared the antibacterial effectivity of the eggs of six precocial and four altricial bird species using Escherichia coli, based on their eggshell traits. The ultrastructure of eggshell was observed using a scanning electron microscope (SEM). According to SEM results, eggs from precocial birds (chicken, turkey, quail, duck, ostrich, and goose) had cuticle on the eggshells, while eggs from altricial birds (pigeon, budgerigar, munia, and canary) did not. The environment/selection pressure may induce the divergent evolution process in eggs of precocial and altricial birds. The E. coli experiment results showed that chicken, turkey, quail, duck, and goose eggs, with a high cuticle opacity, exhibited a much lower E. coli penetration rate. In contrast, the eggs with poor (ostrich) or without (pigeon, budgerigar, munia, and canary) cuticle exhibited a higher penetration rate. It is suggested that cuticle is a main barrier against bacterial penetration in precocial birds' eggs. Turkey and quail eggs showed the lowest E. coli contamination rate (3.33% and 2.22%, respectively), probably because of the tightly connected nanosphere structure on their cuticle. As for altricial birds' eggs, the eggs of budgerigar, munia, and canary with small pore diameter (0.57 to 1.22 µm) had a lower E. coli penetration rate than pigeon eggs (45.56%, 66.67%, 50%, and 97.78%, respectively, P < 0.05), indicating that pore diameter played a significant role in defending against bacterial trans-shell invasion. We found that eggshell thickness and pore area decreased with egg size. The cuticle quality had no relationship with egg size, but was closely related to the bird species. The E. coli penetration rate of altricial birds' eggs was significantly higher than that of precocial birds' eggs, mainly because the pores are exposed on the eggshell surface and cuticle protection is absent. This study provides detailed information on the eggshell cuticle, which gives insight into the cuticle evolution process that occurred in precocial and altricial bird species. Moreover, the results of E. coli penetration may help understanding the antibacterial behavior in birds.


Assuntos
Antibacterianos/farmacologia , Casca de Ovo/microbiologia , Aves Domésticas/microbiologia , Animais , Casca de Ovo/química , Casca de Ovo/ultraestrutura , Escherichia coli/efeitos dos fármacos , Escherichia coli/patogenicidade , Aves Domésticas/fisiologia , Reprodução
17.
Curr Microbiol ; 76(10): 1130-1137, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31342082

RESUMO

Escherichia coli O157:H7 and Salmonella enterica subsp. enterica are the pathogens that frequently cause foodborne illness. Bacteriophage applications have been proposed as effective for preventing food contamination caused by these pathogenic bacteria. Escherichia phage KIT03 was isolated from the soil of a poultry farm in Kyoto, Japan. KIT03 can infect Escherichia coli O157:H7 and Salmonella enterica serotypes Choleraesuis and Enteritidis. One-step growth analysis revealed that KIT03 can propagate within its initial host (E. coli NBRC 3972), E. coli O157:H7 and S. Choleraesuis with an approximate burst size of 39, 51 and 37 phage particles per infected cell, respectively. The morphological type and genome annotation suggested that KIT03 belongs to the family Myoviridae, subfamily Tevenvirinae, genus Tequatrovirus. In vitro challenge tests demonstrated that KIT03 can lyse the tested bacteria and suppress their growth. Based on the susceptibility test and adsorption assay of KIT03 with E. coli K-12 BW25113 mutants, it was proposed that KIT03 may recognise and infect bacteria with a deficient outer core of lipopolysaccharides.


Assuntos
Escherichia coli O157/virologia , Myoviridae/isolamento & purificação , Myoviridae/fisiologia , Salmonella enterica/virologia , Animais , Escherichia coli/genética , Escherichia coli/virologia , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Genoma Viral/genética , Japão , Lipopolissacarídeos/genética , Myoviridae/classificação , Myoviridae/genética , Filogenia , Aves Domésticas/microbiologia
18.
Food Microbiol ; 83: 193-199, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31202413

RESUMO

The fastidious requirement of the zoonotic pathogen Campylobacter jejuni contrasts with its ability to overcome harsh conditions. Different strategies might be involved in the survival and persistence of C. jejuni through the poultry food chain. Therefore, the aims of this study were to get insights in the survival strategies in the poultry slaughterhouse environment by (i) characterizing factors such as biofilm formation, virulence and antimicrobial resistance in environmental isolates and (ii) understanding the possible link between the phenotypic and genetic characterization using whole genome sequencing (WGS). Results have shown that three STs: ST 443 (PFGE A), ST 904 (PFGE C) and ST 3769 (PFGE G), out of the six studied, formed biofilms with variable intensity according to different conditions (temperatures -37 °C, 30 °C, 25°C- and materials -stainless steel and plastic-). High levels of antimicrobial resistance were found in isolates to ciprofloxacin, nalidixic acid and tetracycline as well as to two common detergents used in the slaughterhouse. A combination of several changes in the genome of ST 904 (PFGE C) including mutations, insertions in antimicrobial resistance genes, the presence of T6SS and a set of genes related to virulence factors might explain its ability to form biofilm and persist longer in the environment. However, the complexity of the survival strategies adopted by the different strains of C. jejuni suggests that multiple mechanisms may exist that allow these organisms to persist and ultimately cause disease in humans.


Assuntos
Matadouros , Antibacterianos/farmacologia , Biofilmes/crescimento & desenvolvimento , Infecções por Campylobacter/veterinária , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/patogenicidade , Aves Domésticas/microbiologia , Animais , Técnicas de Tipagem Bacteriana , Campylobacter jejuni/genética , Ciprofloxacino/farmacologia , Farmacorresistência Bacteriana/genética , Microbiologia de Alimentos , Genoma Bacteriano , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Fatores de Virulência/genética , Sequenciamento Completo do Genoma
19.
Vet Microbiol ; 233: 118-123, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31176397

RESUMO

The vast capacity for maintenance and dissemination in the environment are major challenges for the control of Salmonella spp. in poultry farms. The aim of this study was to assess environmental contamination by non-typhoidal Salmonella in successive broiler flocks in nine commercial broiler farms integrated with three companies in the south of Brazil, for a twelve-month production period. Recycled broiler litter, feed and swabs from the evaporative cooling system pads were analyzed, and the total enterobacteria count in the litter samples was ascertained. Positive broiler houses were identified in two of the three broiler companies studied, in which non-typhoidal Salmonella were detected for the first time in the first or second flock, and recurred in the recycled litter of subsequent flocks. Feed and evaporative cooling pad swab samples were also positive in at least one of the assessed flocks. The majority of the isolates (87.5%) originating from different flocks, broiler houses and companies that were sampled were identified as S. Heidelberg, with the prevalence of one single genotype. The total enterobacteria levels in the litter diminished as the flocks progressed, but the presence of Salmonella spp. was constant over the course of time, indicating that the litter management procedures were not capable of interrupting the cycle of residual contamination. The predominance of S. Heidelberg highlights its emergence and dissemination in this region, as well as its resistance and maintenance in the environment, and reinforces the need to improve prevention and recycled litter management measures.


Assuntos
Galinhas/microbiologia , Microbiologia Ambiental , Aves Domésticas/microbiologia , Salmonelose Animal/epidemiologia , Salmonella/genética , Criação de Animais Domésticos , Animais , Brasil/epidemiologia , Genótipo , Abrigo para Animais/normas , Estudos Longitudinais , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/prevenção & controle , Prevalência , Salmonella/isolamento & purificação , Sorogrupo
20.
Vet Microbiol ; 233: 52-60, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31176413

RESUMO

The spread of extended-spectrum ß-lactamases (ESBLs) in Escherichia coli is a major public health issue and ESBL-producing bacteria are frequently reported in livestock. For the assessment of the role of the foodborne transmission pathway in Germany, detailed data on the prevalence and characteristics of isolates of food origin are necessary. The objective of this study was to describe the prevalence of cefotaxime resistant E. coli as well as ESBL/pAmpC-producing E. coli and their characteristics in foods in Germany. Out of 2256 food samples, the highest prevalence of cefotaxime resistant E. coli was observed in chicken meat (74.9%), followed by turkey meat (40.1%). Prevalence in beef, pork and minced meat was considerably lower (4.2-15.3%). Whereas 18.0% of the raw milk samples, collected at farm level were positive, this was true only for few cheese samples (1.3%). In one out of 399 vegetable samples a cefotaxime-resistant E. coli was isolated. ESBL resistance genes of the CTX-M-group (10.1% of all samples) were most frequently detected, followed by genes of the pAmpC (2.6%), SHV (2.0%) and TEM (0.8%) families. Distribution of ESBL/AmpC-encoding E. coli resistance genes and E. coli phylogroups was significantly different between the chicken related food samples and all other food items. Our study results reflect that consumers might get exposed to ESBL/pAmpC-producing E. coli through several food chains. These results together with those collected at primary production and in the human population in other studies will allow more detailed analysis of the foodborne pathways, considering transmission from livestock populations to food at retail and to consumers in Germany.


Assuntos
Proteínas de Bactérias/genética , Escherichia coli/genética , Microbiologia de Alimentos , Carne/microbiologia , beta-Lactamases/genética , Animais , Antibacterianos/farmacologia , Proteínas de Bactérias/biossíntese , Cefotaxima/farmacologia , Farmacorresistência Bacteriana , Escherichia coli/enzimologia , Infecções por Escherichia coli/transmissão , Infecções por Escherichia coli/veterinária , Proteínas de Escherichia coli/biossíntese , Proteínas de Escherichia coli/genética , Alemanha , Gado/microbiologia , Aves Domésticas/microbiologia , Prevalência , Carne Vermelha/microbiologia , Verduras/microbiologia , beta-Lactamases/biossíntese
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