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1.
J Med Microbiol ; 70(3)2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33475480

RESUMO

Introduction. In May-June 2018, an outbreak of campylobacteriosis involved students and school staff from kindergartens and primary schools in Pescara, southern Italy.Aim. We present details of the epidemiological and microbiological investigation, and the findings of the analytical study, as well as the implemented control measures.Methodology. To identify possible risk factors associated with the observed outbreak, a case control study was conducted using a questionnaire to collect information on the date of symptoms onset, type and duration of symptoms, type of healthcare contact, school attendance, and food items consumed at school lunches during the presumed days of exposure. Attack rates were calculated for each date and school. Logistic regression models were used to estimate the odds ratios of being a case and the odds of illness by food items consumed, respectively. Moreover, we carried out a comparative genomic analysis using whole genome multilocus sequence typing (wgMLST) of Campylobacter jejuni strains isolated during the outbreak investigation to identify the source of the outbreak.Results. Overall, 222 probable cases from 21 schools were identified, and C. jejuni was successfully isolated from 60 patients. The meals in the schools involved were provided by two cooking centres managed by a joint venture between two food companies. Environmental and food sampling, epidemiological and microbiological analyses, as well as a case control study with 176 cases and 62 controls from the same schools were performed to identify the source of the outbreak. The highest attack rate was recorded among those having lunch at school on 29 May (7.8 %), and the most likely exposure was 'caciotta' cheese (odds ratio 2.40, 95 % confidence interval 1.10-5.26, P=0.028). C. jejuni was isolated from the cheese, and wgMLST showed that the human and cheese isolates belonged to the same genomic cluster, confirming that the cheese was the vehicle of the infection.Conclusion. It is plausible that a failure of the pasteurization process contributed to the contamination of the cheese batches. Timely suspension of the catering service and summer closure of the schools prevented further spread.


Assuntos
Infecções por Campylobacter , Campylobacter jejuni/isolamento & purificação , Queijo/microbiologia , Surtos de Doenças , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Adulto , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/microbiologia , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Doenças Transmitidas por Alimentos/epidemiologia , Humanos , Itália , Masculino , Pasteurização , Instituições Acadêmicas , Inquéritos e Questionários
2.
BMC Infect Dis ; 21(1): 108, 2021 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-33485317

RESUMO

BACKGROUND: Campylobacter jejuni (C. jejuni) is a leading cause of acute gastroenteritis in human worldwide. The aim of study was to assess the distribution of sialylated lipooligosaccharide (LOS) classes and capsular genotypes in C. jejuni isolated from Iranian children with gastroenteritis. Furthermore, the level of dnaK gene expression in C. jejuni strains with selected capsular genotypes and LOS classes was intended. Moreover, a comprehensive study of C. jejuni MLST-genotypes and inclusive comparison with peer sequences worldwide was intended. METHODS: Twenty clinical C. jejuni strains were isolated from fecal specimens of 280 children aged 0-5 years, suspected of bacterial gastroenteritis, which admitted to 3 children hospitals from May to October, 2018. Distribution of sialylated LOS classes and specific capsular genotypes were investigated in C. jejuni of clinical origin. The expression of dnaK in C. jejuni strains was measured by Real-Time-PCR. MLST-genotyping was performed to investigate the clonal relationship of clinical C. jejuni strains and comparison with inclusive sequences worldwide. RESULTS: C. jejuni HS23/36c was the predominant genotype (45%), followed by HS2 (20%), and HS19 and HS4 (each 10%). A total of 80% of isolates were assigned to LOS class B and C. Higher expression level of dnaK gene was detected in strains with HS23/36c, HS2 and HS4 capsular genotypes and sialylated LOS classes B or C. MLST analysis showed that isolates were highly diverse and represented 6 different sequence types (STs) and 3 clonal complexes (CCs). CC21 and CC257 were the most dominant CCs (75%) among our C. jejuni strains. No new ST and no common ST with our neighbor countries was detected. CONCLUSIONS: The C. jejuni isolates with LOS class B or C, and capsular genotypes of HS23/36, HS2, HS4 and HS19 were dominant in population under study. The CC21 and CC257 were the largest CCs among our isolates. In overall picture, CC21 and CC353 complexes were the most frequently and widely distributed clonal complexes worldwide, although members of CC353 were not detected in our isolates. This provides a universal picture of movement of dominant Campylobacter strains worldwide.


Assuntos
Infecções por Campylobacter/epidemiologia , Campylobacter jejuni/genética , Gastroenterite/epidemiologia , Cápsulas Bacterianas/genética , Proteínas de Bactérias/genética , Infecções por Campylobacter/microbiologia , Campylobacter jejuni/classificação , Campylobacter jejuni/isolamento & purificação , Pré-Escolar , Gastroenterite/microbiologia , Variação Genética , Genótipo , Humanos , Lactente , Recém-Nascido , Irã (Geográfico)/epidemiologia , Lipopolissacarídeos/genética , Tipagem de Sequências Multilocus , Prevalência
3.
BMC Infect Dis ; 21(1): 109, 2021 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-33485326

RESUMO

BACKGROUND: Diarrhoea is a common cause of mortality and morbidity in children under five years old. In Kenya, it has a 21% case fatality with Enteropathogenic E. coli, Campylobacter jejuni, Shigella spp. and Salmonella spp. accounting for 50-60% of the cases. Sulphonamides, tetracycline, ampicillin and trimethoprim/sulfamethoxazole are typically used in the treatment of diarrhoeal diseases but have become ineffective in the face of emerging antimicrobial resistance. The objective of this study was to evaluate the prevalence and antimicrobial susceptibility of Campylobacter jejuni and Shigella species in children under five years of age presenting with diarrhoea at Kapsabet County Referral Hospital in Kenya. METHODS: Faecal samples were collected from 139 children admitted with diarrhoea. Each sample was examined macroscopically for colour, texture, and presence of extraneous material. The samples were then cultured for bacterial growth. Observed bacterial growth was isolated and identified by a series of biochemical tests. Resistance patterns were also evaluated using the Kirby - Bauer Disk diffusion method. The chi - square test and Pearson Correlation Coefficient were used to establish statistical significance. RESULTS: Approximately 33.1% of the total faecal samples tested were positive for enteric pathogens. Shigella spp. demonstrated resistance to erythromycin (91.7%), doxycyclin (83.3%), ampicillin (82.1%), cotrimoxazole (73.1%), minocycline (66.7%) and cefuroxime (54.2%). Campylobacter jejuni also exhibited resistance to erythromycin (87.5%), doxycyclin (75%), ampicillin (73.7%), cotrimoxazole (73.3%) and minocycline (68.8%). CONCLUSIONS: The resistance patterns of Shigella spp. and Campylobacter jejuni reported in this study necessitates the need for a comprehensive multiregional investigation to evaluate the geographical prevalence and antimicrobial resistance distributions of these microorganisms. These findings also support the need for the discovery and development of effective therapeutic alternatives. TRIAL REGISTRATION: Retrospectively registered. Certificate No. 00762.


Assuntos
Antibacterianos/farmacologia , Campylobacter jejuni/efeitos dos fármacos , Diarreia/microbiologia , Farmacorresistência Bacteriana Múltipla , Shigella/efeitos dos fármacos , Antibacterianos/uso terapêutico , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/microbiologia , Campylobacter jejuni/isolamento & purificação , Criança , Pré-Escolar , Diarreia/epidemiologia , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Disenteria Bacilar/epidemiologia , Disenteria Bacilar/microbiologia , Fezes/microbiologia , Feminino , Hospitais , Humanos , Lactente , Quênia/epidemiologia , Masculino , Testes de Sensibilidade Microbiana , Shigella/isolamento & purificação
4.
Int J Food Microbiol ; 339: 109014, 2021 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-33333444

RESUMO

The objective of this study was to develop a method with improved sensitivity for Campylobacter jejuni detection in foods. Nitrogen-doped carbon nanodots (N-CNDs) were synthesized and added to an enrichment medium (Bolton broth) at a concentration of 10 mg/mL. A light-emitting diode (LED) at a wavelength of 425 nm was used to irradiate the N-CNDs-supplemented enrichment medium to induce an exothermic reaction for 1 h. Additionally, a monoclonal antibody specific to C. jejuni NCTC11168 was developed using hybridoma cells to aid detection. The C. jejuni detection capabilities of N-CNDs-supplemented enrichment medium and the conventional Bolton broth enrichment, were compared using duck samples. C. jejuni in the enrichment was detected with the monoclonal antibody based-indirect enzyme-linked immunosorbent assay (ID-ELISA). The N-CNDs-supplemented enrichment medium showed a better C. jejuni detection capability than the conventional Bolton broth enrichment. Additionally, data from ID-ELISA showed excellent detection efficiency and a shortened detection time in the N-CNDs-supplemented enrichment medium after LED irradiation at 425 nm. These results indicate that 1-h LED irradiation at 425 nm to Bolton broth supplemented with the N-CNDs increased the detection efficiency and shortened the detection time with the monoclonal antibody for C. jejuni in food.


Assuntos
Campylobacter jejuni/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Microbiologia de Alimentos/métodos , Carne/microbiologia , Nanopartículas/química , Animais , Anticorpos Monoclonais/metabolismo , Bactérias/efeitos dos fármacos , Carbono/química , Carbono/farmacologia , Meios de Cultura , Patos/microbiologia , Nitrogênio/química , Nitrogênio/farmacologia
5.
PLoS One ; 15(7): e0236889, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32730330

RESUMO

Australian rates of campylobacteriosis are among the highest in developed countries, yet only limited work has been done to characterize Campylobacter spp. in Australian retail products. We performed whole genome sequencing (WGS) on 331 C. coli and 285 C. jejuni from retail chicken meat, as well as beef, chicken, lamb and pork offal (organs). Campylobacter isolates were highly diverse, with 113 sequence types (STs) including 38 novel STs, identified from 616 isolates. Genomic analysis suggests very low levels (2.3-15.3%) of resistance to aminoglycoside, beta-lactam, fluoroquinolone, macrolide and tetracycline antibiotics. A majority (>90%) of isolates (52/56) possessing the fluoroquinolone resistance-associated T86I mutation in the gyrA gene belonged to ST860, ST2083 or ST7323. The 44 pork offal isolates were highly diverse, representing 33 STs (11 novel STs) and harboured genes associated with resistance to aminoglycosides, lincosamides and macrolides not generally found in isolates from other sources. Prevalence of multidrug resistant genotypes was very low (<5%), but ten-fold higher in C. coli than C. jejuni. This study highlights that Campylobacter spp. from retail products in Australia are highly genotypically diverse and important differences in antimicrobial resistance exist between Campylobacter species and animal sources.


Assuntos
Antibacterianos/farmacologia , Infecções por Campylobacter/microbiologia , Campylobacter coli/genética , Campylobacter jejuni/genética , Farmacorresistência Bacteriana/genética , Carne/análise , Animais , Infecções por Campylobacter/tratamento farmacológico , Infecções por Campylobacter/genética , Campylobacter coli/efeitos dos fármacos , Campylobacter coli/isolamento & purificação , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/isolamento & purificação , Bovinos , Galinhas , DNA Bacteriano/genética , Microbiologia de Alimentos , Testes de Sensibilidade Microbiana , Carne Vermelha , Ovinos , Suínos , Sequenciamento Completo do Genoma
6.
PLoS One ; 15(7): e0235841, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32645064

RESUMO

The reservoir and source of human campylobacteriosis is primarily considered to be poultry, but also other such as ruminants, pets and environmental sources are related with infection burden. Multilocus sequence typing is often used for Campylobacter epidemiological studies to determine potential sources of human infections. The collection of 420 Campylobacter jejuni isolates with assigned MLST genotype from poultry (n = 139), cattle (n = 48) and wild birds (n = 101) were used in source attribution analysis. Asymmetric island model with accurate and congruent self-attribution results, was used to determine potential sources of human C. jejuni infections (n = 132) in Baltic States. Source attribution analysis revealed that poultry (88.3%) is the main source of C. jejuni human infections followed by cattle and wild bird with 9.4% and 2.3%, respectively. Our findings demonstrated that clinical cases of C. jejuni infections in Baltic countries are mainly linked to poultry, but also to cattle and wild bird sources.


Assuntos
Aves/microbiologia , Infecções por Campylobacter/microbiologia , Campylobacter jejuni/isolamento & purificação , Bovinos/microbiologia , Aves Domésticas/microbiologia , Animais , Animais Selvagens/microbiologia , Países Bálticos/epidemiologia , Infecções por Campylobacter/epidemiologia , Campylobacter jejuni/genética , Reservatórios de Doenças/microbiologia , Humanos , Tipagem de Sequências Multilocus
7.
BMC Infect Dis ; 20(1): 518, 2020 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-32677920

RESUMO

BACKGROUND: Campylobacter jejuni is a leading cause of bacterial diarrhea worldwide, and increasing rates of fluoroquinolone (FQ) resistance in C. jejuni are a major public health concern. The rapid detection and tracking of FQ resistance are critical needs in developing countries, as these antimicrobials are widely used against C. jejuni infections. Detection of point mutations at T86I in the gyrA gene by real-time polymerase chain reaction (RT-PCR) is a rapid detection tool that may improve FQ resistance tracking. METHODS: C. jejuni isolates obtained from children with diarrhea in Peru were tested by RT-PCR to detect point mutations at T86I in gyrA. Further confirmation was performed by sequencing of the gyrA gene. RESULTS: We detected point mutations at T86I in the gyrA gene in 100% (141/141) of C. jejuni clinical isolates that were previously confirmed as ciprofloxacin-resistant by E-test. No mutations were detected at T86I in gyrA in any ciprofloxacin-sensitive isolates. CONCLUSIONS: Detection of T86I mutations in C. jejuni is a rapid, sensitive, and specific method to identify fluoroquinolone resistance in Peru. This detection approach could be broadly employed in epidemiologic surveillance, therefore reducing time and cost in regions with limited resources.


Assuntos
Infecções por Campylobacter/diagnóstico , Campylobacter jejuni/genética , DNA Girase/genética , Farmacorresistência Bacteriana/genética , Fluoroquinolonas/uso terapêutico , Mutação Puntual , Reação em Cadeia da Polimerase em Tempo Real/métodos , Substituição de Aminoácidos , Infecções por Campylobacter/tratamento farmacológico , Infecções por Campylobacter/microbiologia , Campylobacter jejuni/isolamento & purificação , Criança , Ciprofloxacino/uso terapêutico , Análise Mutacional de DNA/métodos , Diarreia/diagnóstico , Diarreia/tratamento farmacológico , Diarreia/microbiologia , Humanos , Isoleucina/genética , Testes de Sensibilidade Microbiana , Peru , Treonina/genética
8.
Int J Food Microbiol ; 327: 108656, 2020 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-32445835

RESUMO

Campylobacter is the world's leading cause of bacterial gastroenteritis, causing nearly 9 million cases of food poisoning in Europe every year. Poultry is considered the main source of Campylobacter infection to humans. The objectives of the study were to determine occurrence of C. jejuni and C. coli in chickens, the antimicrobial resistance, genotypes, and relatedness of the isolates. A total of 177 chicken samples obtained from informal butcher shops (fresh), formal poultry slaughterhouses (refrigerated) and retail market (frozen) were analyzed. Isolation of Campylobacter spp. was conducted according to the ISO 10272-2006 method. Multiplex PCR was used for confirmation and identification of the isolates. The disk diffusion method was used to determine the antimicrobial resistance of the isolates and multilocus sequence typing was used for genotyping. The proportion of samples with Campylobacter spp. was 31.6% among all chicken samples (fresh and refrigerated 47.5%, frozen 0%) C. coli was isolated from 42.4% of chicken samples obtained from butcher shops and from 18.6% of samples obtained in formal slaughterhouses. C. jejuni was isolated from 17.0% of samples obtained in butcher shops and formal slaughterhouses. Campylobacter spp. was not isolated in frozen chicken samples. All tested isolates showed resistance toward ciprofloxacin and susceptibility toward imipenem and all of the isolates were multidrug resistant toward 5 or more antimicrobials. Three sequence types were identified among 10 C. coli isolates and seven sequence types were identified among 10 C. jejuni isolates. Among sequence types, chicken isolates shared similarities of both phenotypic and genetic levels.


Assuntos
Infecções por Campylobacter/veterinária , Campylobacter coli , Campylobacter jejuni , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/microbiologia , Matadouros , Animais , Antibacterianos/farmacologia , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/microbiologia , Campylobacter coli/efeitos dos fármacos , Campylobacter coli/genética , Campylobacter coli/isolamento & purificação , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/genética , Campylobacter jejuni/isolamento & purificação , Galinhas/microbiologia , Resistência Microbiana a Medicamentos , Doenças Transmitidas por Alimentos/microbiologia , Genótipo , Humanos , Jordânia/epidemiologia , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Reação em Cadeia da Polimerase Multiplex , Aves Domésticas/microbiologia , Prevalência
9.
Poult Sci ; 99(5): 2690-2695, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32359606

RESUMO

Chicken is a leading source of thermotolerant Campylobacter, which triggers human foodborne enteritis. This study evaluated thermotolerant Campylobacter contamination of retail chicken in southern Brazil, using qualitative and quantitative analyses. Selective enrichment in Bolton broth for 24 and 48 h after plating onto modified charcoal-cefoperazone-deoxycholate (mCCD) agar and Preston agar was assessed. The combined results of the detection and enumeration methods revealed a frequency of 70% occurrence of thermotolerant Campylobacter in chicken samples. Campylobacter was enumerated in 60% of the samples, whereas 46% of the samples were positive in the qualitative analysis. Quantitative analysis showed average counts of 3.10 ± 0.15 log10 CFU/sample. Higher numbers of Campylobacter-positive samples were found using 24-h enrichment before plating onto Preston agar (46%) than onto mCCD agar (2%). The majority of isolated strains were identified as Campylobacter jejuni, and Campylobacter coli was also found but to a lesser extent. Subtyping revealed a clear distinction between strains isolated from different chicken sources. The enriched samples plated onto mCCD agar showed extensive spreading of nonproducing extended-spectrum ß-lactamases Proteus mirabilis that hampered the identification of Campylobacter colonies. P. mirabilis strains showed resistance to cefoperazone, trimethoprim, and polymyxin B present in broth and plate media used and were inhibited by rifampicin present in Preston agar. The results underline the effect of the spread of contaminant strains on Campylobacter cultures, which might be prevented using a recently revised International Organization for Standardization method for qualitative analysis of chicken.


Assuntos
Campylobacter coli/isolamento & purificação , Campylobacter jejuni/isolamento & purificação , Microbiologia de Alimentos , Carne/microbiologia , Termotolerância , Animais , Brasil , Campylobacter coli/fisiologia , Campylobacter jejuni/fisiologia , Galinhas
10.
Int J Food Microbiol ; 326: 108641, 2020 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-32371295

RESUMO

Thermotolerant Campylobacter is the leading bacterial cause of foodborne illness in humans worldwide. The objectives of this study were to estimate prevalence and to identify and characterize potential sources of thermotolerant Campylobacter contamination in broilers on farms and at the slaughterhouse; to evaluate the clonal relationship among thermotolerant Campylobacter isolates from different stages of the broiler meat supply chain, and to analyze the presence of virulence genes in different sources of thermotolerant Campylobacter. A total of 1210 samples were collected from three broiler meat supply chains in Santa Fe, Argentina. At the farms, the sampling collection included broilers one week prior to slaughter, wild-living birds, domestic dogs, wild rodents, farm workers' boots, litter, feed, drinking water, flies, and darkling beetles (Alphitobius diaperinus). At the slaughtering line, the samples taken were from the evisceration zone (broiler cecum, working surfaces, evisceration knives and workers' hands), from the chiller zone (surfaces and direct supply water) and from the packing zone (work surfaces, workers' hands and broiler carcasses). The samples taken along each supply chain were in the same batch. The isolates obtained were identified to the species level (C. jejuni and C. coli) by multiplex PCR and were analyzed using pulsed-field gel electrophoresis to compare different profiles according to the source. Finally, the presence of 11 virulence genes was examined (cadF, cdtA, cdtB, cdtC, ciaB, flaA, flhA, iam, wlaN, virB11, racR). From 254 isolates, 128 (50.4%) were Campylobacter jejuni and 126 (49.6%) Campylobacter coli. C. jejuni was the species most prevalent in farm and C. coli the species most prevalent at the slaughterhouse. We detected thermotolerant Campylobacter in samples of wild birds, darkling beetles, farm workers' boots, flies and litter. At the slaughterhouse, the prevalence varied along the process line. By analyzing PFGE results, C. jejuni showed 21 profiles with three predominant genotypes, while C. coli showed 14 profiles with four predominant genotypes. A high genotype diversity was found; however, relationships between isolates from different stages of the broiler meat chain, between broiler and potential sources of thermotolerant Campylobacter contamination and between strains in the farm and in the slaughterhouse were detected. Furthermore, there was evidence of cross-contamination at the slaughterhouse. FlaA, flhA genes were detected in all strains, and the third most prevalent virulence gene was cadF. Only those strains obtained from flies, wild-living birds and broiler carcass samples harbored 10 of 11 pathogenic genes. The prevalence of some pathogenic genes between C. jejuni and C. coli was different. This evidence should contribute the scientific basis to implement risk management measures in public health.


Assuntos
Campylobacter coli/genética , Campylobacter jejuni/genética , Doenças Transmitidas por Alimentos/microbiologia , Carne/microbiologia , Aves Domésticas/microbiologia , Matadouros/estatística & dados numéricos , Animais , Argentina , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Bactérias/genética , Infecções por Campylobacter/microbiologia , Campylobacter coli/isolamento & purificação , Campylobacter coli/patogenicidade , Campylobacter jejuni/isolamento & purificação , Campylobacter jejuni/patogenicidade , Proteínas de Transporte/genética , Ceco/microbiologia , Galinhas/microbiologia , Besouros/microbiologia , Dípteros/microbiologia , Cães , Água Potável/microbiologia , Eletroforese em Gel de Campo Pulsado , Flagelina/genética , Genótipo , Humanos , Indústria de Embalagem de Carne/estatística & dados numéricos , Proteínas de Membrana/genética , Prevalência , Roedores/microbiologia , Termotolerância , Virulência/genética
11.
New Microbiol ; 43(2): 96-98, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32310303

RESUMO

Infections caused by Campylobacter jejuni are rarely associated with extraintestinal complications. C. jejuni bacteremia is difficult to detect in patients with hematological malignancies undergoing chemotherapy where the choice of appropriate antibiotic treatment is extremely important. We report two cases of C. jejuni bacteremia in Italian pediatric patients affected by acute lymphoblastic leukemia (ALL). Agreeing with the most recent epidemiological data, both clinical isolates showed a typical phenotypic antimicrobial resistance patterns with combined resistance to ciprofloxacin and tetracycline. To our knowledge, this is the first report of C. jejuni isolation from the blood of ALL pediatric patients in Italy, and it provides important epidemiological information on this rare infection.


Assuntos
Bacteriemia , Infecções por Campylobacter , Campylobacter jejuni , Leucemia-Linfoma Linfoblástico de Células Precursoras , Antibacterianos , Bacteriemia/complicações , Infecções por Campylobacter/complicações , Infecções por Campylobacter/diagnóstico , Campylobacter jejuni/isolamento & purificação , Criança , Farmacorresistência Bacteriana , Humanos , Itália , Testes de Sensibilidade Microbiana , Leucemia-Linfoma Linfoblástico de Células Precursoras/complicações
12.
PLoS One ; 15(4): e0231810, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32315369

RESUMO

Campylobacter jejuni has caused several campylobacteriosis outbreaks via raw milk consumption. This study reports follow-up of a milk-borne campylobacteriosis outbreak that revealed persistent C. jejuni contamination of bulk tank milk for seven months or longer. Only the outbreak-causing strain, representing sequence type (ST) 883, was isolated from milk, although other C. jejuni STs were also isolated from the farm. We hypothesized that the outbreak strain harbors features that aid its environmental transmission or survival in milk. To identify such phenotypic features, the outbreak strain was characterized for survival in refrigerated raw milk and in aerobic broth culture by plate counting and for biofilm formation on microplates by crystal violet staining and quantification. Furthermore, whole-genome sequences were studied for such genotypic features. For comparison, we characterized isolates representing other STs from the same farm and an ST-883 isolate that persisted on another dairy farm, but was not isolated from bulk tank milk. With high inocula (105 CFU/ml), ST-883 strains survived in refrigerated raw milk longer (4-6 days) than the other strains (≤3 days), but the outbreak strain showed no outperformance among ST-883 strains. This suggests that ST-883 strains may share features that aid their survival in milk, but other mechanisms are required for persistence in milk. No correlation was observed between survival in refrigerated milk and aerotolerance. The outbreak strain formed a biofilm, offering a potential explanation for persistence in milk. Whether biofilm formation was affected by pTet-like genomic element and phase-variable genes encoding capsular methyltransferase and cytochrome C551 peroxidase warrants further study. This study suggests a phenotypic target candidate for interventions and genetic markers for the phenotype, which should be investigated further with the final aim of developing control strategies against C. jejuni infections.


Assuntos
Infecções por Campylobacter/epidemiologia , Campylobacter jejuni/isolamento & purificação , Microbiologia de Alimentos , Leite/microbiologia , Alimentos Crus/microbiologia , Animais , Campylobacter jejuni/genética , Bovinos , Surtos de Doenças , Fazendas , Fezes/microbiologia , Finlândia , Humanos
13.
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
14.
J Vet Diagn Invest ; 32(3): 413-419, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32274977

RESUMO

Campylobacter infection is a leading cause of ovine abortion worldwide. Campylobacter fetus and C. jejuni are the major species involved. We report herein on abortion storms in 4 Danish sheep flocks. Initially, no pathogenic bacteria were isolated from placental and fetal tissues on aerobic and selective media despite the presence of severe suppurative and necrotizing placentitis with numerous bacteria located intracellularly in trophoblasts. Fluorescence in situ hybridization (FISH) was then applied on abortion material from 13 cases; species-specific oligonucleotide probes directed against either C. fetus or C. jejuni were used in combination with a general bacterial probe. C. fetus was detected as the only lesion-associated bacterial species in 4 cases from 2 flocks, and C. jejuni in 6 cases from the other 2 flocks, thereby establishing the likely etiology of the abortion storms in all 4 flocks. FISH is a useful detection tool in culture-negative cases with tissue lesions suggestive of bacterial infection. Furthermore, FISH is a fast and economical method to detect and identify the zoonotic agent Campylobacter within ovine abortion material.


Assuntos
Aborto Animal/microbiologia , Infecções por Campylobacter/veterinária , Campylobacter fetus/isolamento & purificação , Campylobacter jejuni/isolamento & purificação , Hibridização in Situ Fluorescente/veterinária , Doenças dos Ovinos/diagnóstico , Animais , Infecções por Campylobacter/diagnóstico , Infecções por Campylobacter/microbiologia , Feminino , Ovinos , Doenças dos Ovinos/microbiologia , Especificidade da Espécie
15.
Infection ; 48(3): 471-475, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32128685

RESUMO

BACKGROUND: While Campylobacter jejuni represents the most common cause of bacterial gastroenteritis, Yersinia pseudotuberculosis infections are very rarely diagnosed in adults. CASE: We report on a previously healthy patient who presented several times at our hospital with fever, Guillain-Barré syndrome, recurrent abdominal symptoms and distinct mesenteric lymphadenopathy, respectively. This complicated and diagnostically challenging course of disease was caused by a C. jejuni and Y. pseudotuberculosis coinfection. Antibiotic treatment with doxycycline was effective. CONCLUSION: Broad serology testing was crucial to discover that two concomitant infections were causing the symptoms. This case demonstrates that when a clinical picture is not fully explained by one known infection, another infection with the same underlying risk factor has to be considered, hence "a horse and a zebra".


Assuntos
Infecções por Campylobacter/diagnóstico , Coinfecção/diagnóstico , Gastroenteropatias/diagnóstico , Síndrome de Guillain-Barré/diagnóstico , Infecções por Yersinia pseudotuberculosis/diagnóstico , Adulto , Antibacterianos/uso terapêutico , Infecções por Campylobacter/diagnóstico por imagem , Infecções por Campylobacter/tratamento farmacológico , Infecções por Campylobacter/microbiologia , Campylobacter jejuni/isolamento & purificação , Coinfecção/diagnóstico por imagem , Coinfecção/tratamento farmacológico , Coinfecção/microbiologia , Doxiciclina/uso terapêutico , Febre/microbiologia , Gastroenteropatias/diagnóstico por imagem , Gastroenteropatias/tratamento farmacológico , Gastroenteropatias/microbiologia , Alemanha , Síndrome de Guillain-Barré/diagnóstico por imagem , Síndrome de Guillain-Barré/microbiologia , Humanos , Linfadenopatia/diagnóstico , Linfadenopatia/microbiologia , Masculino , Recidiva , Resultado do Tratamento , Yersinia pseudotuberculosis/isolamento & purificação , Infecções por Yersinia pseudotuberculosis/diagnóstico por imagem , Infecções por Yersinia pseudotuberculosis/tratamento farmacológico , Infecções por Yersinia pseudotuberculosis/microbiologia
16.
Appl Environ Microbiol ; 86(9)2020 04 17.
Artigo em Inglês | MEDLINE | ID: mdl-32111591

RESUMO

Campylobacter spp. have been recognized as major foodborne pathogens worldwide. An increasing frequency of antibiotic-resistant pathogens, including Campylobacter spp., have been identified to transmit from food products to humans and cause severe threats to public health. To better mitigate the antibiotic resistance crisis, rapid detection methods are required to provide timely antimicrobial resistance surveillance data for agri-food systems. Herein, we developed a polymer-based microfluidic device for the identification and antimicrobial susceptibility testing (AST) of Campylobacter spp. An array of bacterial incubation chambers were created in the microfluidic device, where chromogenic medium and antibiotics were loaded. The growth of Campylobacter spp. was visualized by color change due to chromogenic reactions. This platform achieved 100% specificity for Campylobacter identification. Sensitive detection of multiple Campylobacter species (C. jejuni, C. coli, and C. lari) was obtained in artificially contaminated milk and poultry meat, with detection limits down to 1 × 102 CFU/ml and 1 × 104 CFU/25 g, respectively. On-chip AST determined Campylobacter antibiotic susceptibilities by the lowest concentration of antibiotics that can inhibit bacterial growth (i.e., no color change observed). High coincidences (91% to 100%) of on-chip AST and the conventional agar dilution method were achieved against several clinically important antibiotics. For a presumptive colony, on-chip identification and AST were completed in parallel within 24 h, whereas standard methods, including biochemical assays and traditional culture-based AST, take several days for multiple sequential steps. In conclusion, this lab-on-a-chip device can achieve rapid and reliable detection of antibiotic-resistant Campylobacter spp.IMPORTANCE Increasing concerns of antibiotic-resistant Campylobacter spp. with regard to public health emphasize the importance of efficient and fast detection. This study described the timely identification and antimicrobial susceptibility testing of Campylobacter spp. by using a microfluidic device. Our developed method not only reduced the total analysis time, but it also simplified food sample preparation and chip operation for end users. Due to the miniaturized size of the lab-on-a-chip platform, the detection was achieved by using up to 1,000 times less of the reagents than with standard reference methods, making it a competitive approach for rapid screening and surveillance study in food industries. In addition, multiple clinically important Campylobacter species (C. jejuni, C. coli, and C. lari) could be tested by our device. This device has potential for wide application in food safety management and clinical diagnostics, especially in resource-limited regions.


Assuntos
Campylobacter coli/efeitos dos fármacos , Campylobacter jejuni/efeitos dos fármacos , Campylobacter lari/efeitos dos fármacos , Farmacorresistência Bacteriana Múltipla , Testes de Sensibilidade Microbiana/métodos , Microfluídica/métodos , Antibacterianos/farmacologia , Campylobacter coli/isolamento & purificação , Campylobacter jejuni/isolamento & purificação , Campylobacter lari/isolamento & purificação , Dispositivos Lab-On-A-Chip
17.
BMC Infect Dis ; 20(1): 199, 2020 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-32143636

RESUMO

BACKGROUND: Campylobacter jejuni is a motile, gram-negative rod known for causing self-limiting enterocolitis while rarely causing extraintestinal infections. We report the first case of a patient with Campylobacter jejuni osteomyelitis in both femora. CASE PRESENTATION: A 54-year-old female presented with progressive pain in both upper extremities. Her past medical history mentioned a lymphoplasmacytic lymphoma (LPL) for which she had received dexamethasone, cyclophosphamide and fludarabine and was still receiving maintenance therapy with intravenous rituximab. Two months prior to presentation, she received oral fluoroquinolone for a recurrent enterocolitis with stool cultures positive for Campylobacter jejuni. Palpation of the left quadriceps femoris muscle was remarkably painful during physical examination. Laboratory testing showed elevated C-reactive protein and recent low gamma globulin levels. The presumptive diagnosis at this point was a transformation of LPL to a large B cell lymphoma. In order to determine the preferred site for biopsy, a fluorine-18 fluoro-2-deoxy-D-glucose positron emission tomography combined with computed tomography was done. However, blood cultures taken on admission showed growth of Campylobacter jejuni in both aerobic bottles, with a strain resistant to fluoroquinolones. Diagnosis of Campylobacter jejuni osteomyelitis was confirmed with 16S ribosomal RNA gene polymerase chain reaction performed on femoral bone obtained through biopsy. Treatment with intravenous imipenem/cilastatin followed by intravenous and oral doxycycline proved insufficient. Subsequently, the patient was treated successfully with intravenous meropenem for six weeks and concurrent intravenous immunoglobulin. CONCLUSION: We report the first case of Campylobacter jejuni osteomyelitis in both femora in a patient with acquired hypogammaglobulinemia. Diagnosis was confirmed by blood cultures and positive 16S ribosomal RNA gene polymerase chain reaction for Campylobacter spp. on bone biopsy. Treatment was successful with intravenous meropenem and immunoglobulin. Our report showcases an unusual manifestation in a patient with immunodeficiency and discusses failure of initial antibiotic therapy.


Assuntos
Infecções por Campylobacter/diagnóstico , Imunodeficiência de Variável Comum/diagnóstico , Osteomielite/diagnóstico , Antibacterianos/uso terapêutico , Proteína C-Reativa/análise , Infecções por Campylobacter/complicações , Infecções por Campylobacter/tratamento farmacológico , Infecções por Campylobacter/microbiologia , Campylobacter jejuni/genética , Campylobacter jejuni/isolamento & purificação , Imunodeficiência de Variável Comum/complicações , Doxiciclina/uso terapêutico , Feminino , Fêmur/efeitos dos fármacos , Humanos , Pessoa de Meia-Idade , Osteomielite/complicações , Osteomielite/tratamento farmacológico , Osteomielite/microbiologia , Tomografia Computadorizada com Tomografia por Emissão de Pósitrons , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo
18.
PLoS Negl Trop Dis ; 14(3): e0007888, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32182238

RESUMO

BACKGROUND: Campylobacter is a common, but neglected foodborne-zoonotic pathogen, identified as a growing cause of foodborne disease worldwide. Wildlife and domestic animals are considered important reservoirs, but little is known about pathogen infection dynamics in free-ranging mammalian wildlife particularly in sub-Saharan Africa. In countries like Botswana, there is significant overlap between humans and wildlife, with the human population having one of the highest HIV infection rates in the world, increasing vulnerability to infection. METHODOLOGY/PRINCIPAL FINDINGS: We investigated Campylobacter occurrence in archived human fecal samples (children and adults, n = 122, 2011), feces from free-ranging banded mongooses (Mungos mungo, n = 201), surface water (n = 70), and river sediment samples (n = 81) collected in 2017 from the Chobe District, northern Botswana. Campylobacter spp. was widespread in humans (23.0%, 95% CI 13.9-35.4%), with infections dominantly associated with C. jejuni (82.1%, n = 28, 95% CI 55.1-94.5%). A small number of patients presented with asymptomatic infections (n = 6). While Campylobacter spp. was rare or absent in environmental samples, over half of sampled mongooses tested positive (56%, 95% CI 45.6-65.4%). Across the urban-wilderness continuum, we found significant differences in Campylobacter spp. detection associated with the type of den used by study mongooses. Mongooses utilizing man-made structures as den sites had significantly higher levels of C. jejuni infection (p = 0.019) than mongooses using natural dens. Conversely, mongooses using natural dens had overall higher levels of detection of Campylobacter at the genus level (p = 0.001). CONCLUSIONS: These results suggest that landscape features may have important influences on Campylobacter species exposure and transmission dynamics in wildlife. In particular, data suggest that human-modified landscapes may increase C. jejuni infection, a primarily human pathogen, in banded mongooses. Pathogen circulation and transmission in urbanizing wildlife reservoirs may increase human vulnerability to infection, findings that may have critical implications for both public and animal health in regions where people live in close proximity to wildlife.


Assuntos
Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/veterinária , Campylobacter jejuni/isolamento & purificação , Herpestidae/microbiologia , Adolescente , Adulto , Animais , Botsuana/epidemiologia , Infecções por Campylobacter/transmissão , Criança , Pré-Escolar , Transmissão de Doença Infecciosa , Fezes/microbiologia , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Saúde Única , Rios/microbiologia , Adulto Jovem
19.
PLoS One ; 15(1): e0226238, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31978059

RESUMO

Campylobacter jejuni infection is one of the most frequently reported foodborne bacterial diseases worldwide. The main transmission route of these microorganisms to humans is consumption of contaminated food, especially of chicken origin. The aim of this study was to analyze the genetic relatedness of C. jejuni from chicken sources (feces, carcasses, and meat) and from humans with diarrhea as well as to subtype the isolates to gain better insight into their population structure present in Poland. C. jejuni were genotyped using multilocus sequence typing (MLST) and sequence types (STs) were assigned in the MLST database. Among 602 isolates tested, a total of 121 different STs, including 70 (57.9%) unique to the isolates' origin, and 32 STs that were not present in the MLST database were identified. The most prevalent STs were ST464 and ST257, with 58 (9.6%) and 52 (8.6%) C. jejuni isolates, respectively. Isolates with some STs (464, 6411, 257, 50) were shown to be common in chickens, whereas others (e.g. ST21 and ST572) were more often identified among human C. jejuni. It was shown that of 47 human sequence types, 26 STs (106 isolates), 23 STs (102 isolates), and 29 STs (100 isolates) were also identified in chicken feces, meat, and carcasses, respectively. These results, together with the high and similar proportional similarity indexes (PSI) calculated for C. jejuni isolated from patients and chickens, may suggest that human campylobacteriosis was associated with contaminated chicken meat or meat products or other kinds of food cross-contaminated with campylobacters of chicken origin. The frequency of various sequence types identified in the present study generally reflects of the prevalence of STs in other countries which may suggest that C. jejuni with some STs have a global distribution, while other genotypes may be more restricted to certain countries.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Infecções por Campylobacter/microbiologia , Campylobacter jejuni/genética , Diarreia/diagnóstico , Doenças Transmitidas por Alimentos/microbiologia , Carne/análise , Tipagem de Sequências Multilocus/métodos , Animais , Infecções por Campylobacter/epidemiologia , Campylobacter jejuni/classificação , Campylobacter jejuni/isolamento & purificação , Galinhas , Diarreia/genética , Diarreia/microbiologia , Doenças Transmitidas por Alimentos/epidemiologia , Humanos , Carne/microbiologia , Polônia/epidemiologia , Prevalência
20.
PLoS One ; 15(1): e0227535, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31999701

RESUMO

BACKGROUND: Campylobacter jejuni is the most common bacterial cause of human infectious intestinal disease. METHODS: We genome sequenced 601 human C. jejuni isolates, obtained from two large prospective studies of infectious intestinal disease (IID1 [isolates from 1993-1996; n = 293] and IID2 [isolates from 2008-2009; n = 93]), the INTEGRATE project [isolates from 2016-2017; n = 52] and the ENIGMA project [isolates from 2017; n = 163]. RESULTS: There was a significant increase in the prevalence of the T86I mutation conferring resistance to fluoroquinolone between each of the three later studies (IID2, INTEGRATE and ENIGMA) and IID1. Although the distribution of major multilocus sequence types (STs) was similar between the studies, there were changes in both the abundance of minority STs associated with the T86I mutation, and the abundance of clones within single STs associated with the T86I mutation. DISCUSSION: Four population-based studies of community diarrhoea over a 25 year period revealed an increase over time in the prevalence of the T86I amongst isolates of C. jejuni associated with human gastrointestinal disease in the UK. Although associated with many STs, much of the increase is due to the expansion of clones associated with the resistance mutation.


Assuntos
Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/genética , Farmacorresistência Bacteriana/genética , Fluoroquinolonas/farmacologia , Enteropatias/microbiologia , Mutação , Campylobacter jejuni/isolamento & purificação , Campylobacter jejuni/fisiologia , Criança , Genoma Bacteriano/genética , Humanos , Filogenia , Polimorfismo de Nucleotídeo Único , Prevalência , Reino Unido
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