RESUMO
Campylobacter is gram-negative bacteria considered the predominant genera isolated from poultry samples and associated with gastroenteritis. Due to the problems in conventional cultural methods of time-consuming and technically demanding requirements, a rapid and feasible method for their identification and discrimination of the closely related spp. Including Campylobacter coli, Campylobacter fetus, and Campylobacter jejuni is needed. This study analyzes the chicken and sheep meats samples (n = 125) using culture and pre-enrichment-based Quadraplex real-time PCR by targeting OrfA, CstA, HipO, and 16 S rRNA genes of C. coli, C. fetus, C. jejuni and Campylobacter spp. Respectively. The analysis of 125 chicken and sheep meat samples by culture and real-time PCR showed high concordance between the results of the two methods. The present study show high prevalence of Campylobacter species (35% and 32% from chicken and meat respectively) of which C. jejuni were the most abundant. Reaction efficiencies were between 90 and 110%, and detect as low as 8.9 fg in C. jejuni. The need for quick detection and discrimination methods in sheep and chicken meat can be met using the described Quadraplex real-time PCR methodology.
Assuntos
Campylobacter coli , Campylobacter jejuni , Galinhas , Carne , Reação em Cadeia da Polimerase em Tempo Real , Animais , Galinhas/microbiologia , Ovinos/microbiologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Campylobacter coli/genética , Campylobacter coli/isolamento & purificação , Campylobacter coli/classificação , Campylobacter jejuni/genética , Campylobacter jejuni/isolamento & purificação , Campylobacter jejuni/classificação , Carne/microbiologia , Campylobacter fetus/genética , Campylobacter fetus/isolamento & purificação , Campylobacter fetus/classificação , Campylobacter/genética , Campylobacter/isolamento & purificação , Campylobacter/classificação , Microbiologia de Alimentos , DNA Bacteriano/genéticaRESUMO
Cattle farming is a major livestock activity with economic relevance in Rio Grande do Sul (RS), Brazil. However, this activity is still considered of intermediate to low technological level, and in this region, there are few epidemiologic reports of Campylobacter fetus subsp. venerealis (Cfv), the causative agent of bovine genital campylobacteriosis (BGC). Thus, we designed a cross-sectional study to assess the prevalence and Cfv-associated factors in cattle farms in RS, Brazil. In total, 99 farms were randomly selected to participate in the survey. Preputial mucus samples from selected bulls were collected twice (within a 15-day interval) and subjected to Cfv molecular detection. A farm was considered positive when at least one sample was positive for Cfv. Our findings indicate that the farm-level Cfv prevalence in RS is 67.67%. On average, the chance of a farm using natural service to be Cfv-positive increased approximately twice compared to farms that do not use natural service. We also determined that Cfv routine tests reduce the chance of a farm being positive by 92%. Therefore, both Cfv detection tests and the reduction of natural services decrease the chance of a farm being positive for Cfv. Finally, we conclude that Cfv is widely spread in Southern Brazil cattle farms and it is urgent the implementation of control measures to reduce Cfv prevalence in the target population.
Assuntos
Infecções por Campylobacter , Doenças dos Bovinos , Bovinos , Animais , Masculino , Campylobacter fetus , Fazendas , Estudos Transversais , Brasil/epidemiologia , Prevalência , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/veterinária , Infecções por Campylobacter/microbiologia , Doenças dos Bovinos/microbiologiaRESUMO
Bovine genital campylobacteriosis (BGC) is a leading cause of return to estrus in cows. The etiologic agent, Campylobacter fetus subsp. venerealis (Cfv) is transmitted by venereal route. Hence, the surrounding reproductive tract microenvironment could play a role in return to estrus in cows. The presence of Cfv in cervicovaginal mucus of Angus breed females provide three experimental groups, which were subject to bacteriome analyses: 10 Cfv-positive cows (CVP), 10 Cfv-negative cows (CVN), and 10 nonsexually active heifers (NSA). Cows with return to estrus showed higher bacterial richness than NSA. Beta diversity analysis showed a significant difference (P = 0.006) in bacterial composition among the three groups analyzed (CVP, CVN, and NSA). However, no significant difference was found when comparing the CVP versus CVN groups. Ureaplasma and Pseudomonas were the genera most frequently observed in NSA, being Ureaplasma the predictor genus to that group, whereas Alistipes, Bacteroides, Rikenellaceae_RC9_gut_group, UCG-005, and UCG-10 were both significantly more abundant and predictors genera in cows with return to estrus. Our results provide an overview of the cervicovaginal bacterial microbiota in cows harboring Cfv and improve the knowledge of the pathogenesis of BGC.
Assuntos
Infecções por Campylobacter , Campylobacter fetus , Bovinos , Animais , Feminino , Campylobacter fetus/genética , Infecções por Campylobacter/veterinária , Infecções por Campylobacter/microbiologia , MucoRESUMO
Campylobacter fetus is a gram-negative motile bacterium, with two subspecies relevant to cattle health: C. fetus subsp. venerealis (Cfv) and C. fetus subsp. fetus (Cff). Both subspecies are associated with reproductive losses in cattle. In this study, we evaluated the identification of C. fetus for the diagnosis of bovine campylobacteriosis through bacteriological culture, direct immunofluorescence (DIF) and molecular tests in preputial smegma (PS) samples of three Angus bulls challenged with Cfv, Cfv biovar intermedius (Cfvi) or Cff, respectively, in an experiment imitating the natural infection. Two DNA extraction protocols were tested (in-house thermal extraction and commercial kit). Aspiration and scraping collection for PS were compared by conventional tests. Additionally, bacteremia was also evaluated in blood samples. Bulls were challenged by natural mating with heifers that had been experimentally infected with C. fetus subspecies; which led to infection. The Cfv- and Cfvi-bulls were positive for at least 9 months. Although Cff is not considered a venereal strain, in this study it was transmissible to bull from heifers experimentally infected, as evidenced by its colonization and persistence in the preputial cavity for 5 to 6 months. This finding suggests a potential risk of dissemination within herds. The results obtained by bacteriological culture or direct immunofluorescence (DIF) showed no significant differences, regardless the sampling device used (aspiration with Cassou pipette, metal and plastic scraper). C. fetus qPCR, on the other hand, yielded better results with an in-house DNA extraction method than with a commercial kit (75% vs 66.6%). Furthermore, qPCR diagnosis was more efficient than culture (66.6%) or DIF (56%). Bacteremia in whole blood samples was negative by qPCR and bacteriological culture in all samples. Altogether, this study demonstrated the transmission of Cff from heifers to bull and also showed that PCR-based methods are promising for the diagnosis of Bovine Genital Campylobacteriosis from clinical samples of PS.
Assuntos
Infecções por Campylobacter , Doenças dos Bovinos , Doenças Urogenitais , Bovinos , Animais , Masculino , Feminino , Doenças dos Bovinos/microbiologia , Infecções por Campylobacter/microbiologia , Reação em Cadeia da Polimerase , Campylobacter fetus/genéticaRESUMO
We report a rare case of Campylobacter fetus bacteremia in a 50-year-old woman following kidney transplantation. Bacteremia was complicated by multivisceral signs such as multiple splenic abscesses, bacterial hepatitis, erythema nodosum and reactive arthritis. Despite a prolonged diagnostic delay, the diagnosis was made on blood culture identification and the global outcome was favorable with adequate antibiotherapy. Reports in the literature describe a high rate of mortality for Campylobacter spp. septicemia, with most patients being immunocompromised. However, Campylobacter spp. has been rarely described in renal transplant patients. Moreover, a splenic septic localization due to Campylobacter spp. has been reported only once to our knowledge. Clinicians should be aware of the diagnostic difficulties related to the frequent negativity of stool samples in C. fetus septicemia, in order to implement a tailored medical strategy. Some data suggest that rapid introduction of adapted antibiotic therapy is associated with a reduction in mortality.
Assuntos
Bacteriemia , Infecções por Campylobacter , Transplante de Rim , Esplenopatias , Abscesso/diagnóstico , Bacteriemia/diagnóstico , Bacteriemia/tratamento farmacológico , Infecções por Campylobacter/complicações , Infecções por Campylobacter/diagnóstico , Infecções por Campylobacter/tratamento farmacológico , Campylobacter fetus , Diagnóstico Tardio , Feminino , Humanos , Transplante de Rim/efeitos adversos , Pessoa de Meia-Idade , Esplenopatias/complicaçõesRESUMO
Campylobacter fetusfetus (Cff) is a major infectious cause of abortion in sheep worldwide, and an opportunistic human pathogen. Information on Cff as an ovine abortifacient in South America is limited. We describe a case of abortion caused by a multidrug resistant strain of Cff in a sheep in Uruguay. In August 2017, 3/57 pregnant ewes (5.3%) aborted whithin one week. Histopathologic examination of the placenta of an aborted ewe revealed severe neutrophilic and fibrinonecrotizing placentitis with vasculitis and thrombosis of the chorionic arterioles. Cff was isolated on microaerobic culture in Skirrow agar, and further confirmed by 16S rDNA PCR amplification and sequencing, and endpoint and real time PCR assays. Antimicrobial sensitivity testing revealed resistance to tetracyclines, nalidixic acid, telithromycin and clindamycin. Other abortifacients were not detected. Further studies are necessary to determine the geographic distribution, ecology, epidemiology, economic impact, and antimicrobial resistance of Cff in sheep flocks in Uruguay.
Assuntos
Infecções por Campylobacter , Campylobacter , Doenças dos Ovinos , Aborto Animal , Animais , Infecções por Campylobacter/diagnóstico , Infecções por Campylobacter/patologia , Infecções por Campylobacter/veterinária , Campylobacter fetus/genética , Feminino , Feto/patologia , Gravidez , Ovinos , Doenças dos Ovinos/diagnóstico , UruguaiRESUMO
The study of outer membrane vesicles (OMVs) became relevant because of their probable important role in the transfer of virulence factors to host cells. Campylobacter fetus is mainly a mammal pathogen whose virulence characterization is still limited. The aim of this study was to evaluate and to characterize the secretion of OMVs in this bacterium. By transmission electron microscopy, we confirmed the production of OMVs in all the strains assayed. Purified OMVs showed a spherical shape and variable size, although comparable to those of other gram-negative bacteria. We also confirmed the presence of the S-layer on the surface of the OMVs of all the strains assayed with the exception of those derived from the NTCC reference strain. In addition, we demonstrated their immunoreactivity by the dot-blot assay. Hence, C. fetus OMVs could contribute to the modulation of the host response and constitute a candidate to be evaluated as an adjuvant of current vaccines used in the veterinary field. This work represents a platform to drive future studies towards the role of these subcellular structures in C. fetus-host interaction.
Assuntos
Proteínas da Membrana Bacteriana Externa , Campylobacter fetus , Animais , Proteínas da Membrana Bacteriana Externa/química , Bactérias Gram-Negativas , Mamíferos , Virulência , Fatores de VirulênciaRESUMO
The sensitivity (Se) and specificity (Sp) of three diagnostic tests for the detection of Campylobacter fetus venerealis (Cfv) using field samples were estimated using a Bayesian latent class model (BLCM), accounting for the absence of a gold standard. The tests included in this study were direct immunofluorescence antibody test (IFAT), polymerase chain reaction (PCR), and real-time PCR (RT-PCR). Twelve farms from two different populations were selected and bull prepuce samples were collected. The IFAT was performed according to the OIE Manual. The conventional PCR was performed as multiplex, targeting the gene nahE for C. fetus species identification and insertion element ISCfe1 for Cfv identification. The RT-PCR was performed as uniplex: one targeting the gene nahE for C. fetus and the other targeting the insertion ISCfe1 (ISC2) for Cfv. Results from the BLCM showed a median Se of 11.7% (Bayesian credibility interval (BCI): 1.93-29.79%), 53.7% (BCI: 23.1-95.0%), and 36.1% (BCI: 14.5-71.7%) for IFAT, PCR, and RT-PCR respectively. The Sp were 94.5% (BCI: 90.1-97.9%), 97.0% (BCI: 92.9-99.3%), and 98.4% (BCI: 95.3-99.7%) for IFAT, PCR, and RT-PCR respectively. The correlation between PCR and RT-PCR was positive and low in samples from both sampled population (0.63% vs 8.47%). These results suggest that diagnostic sensitivity of the studied tests is lower using field samples than using pure Cfv strains.
Assuntos
Infecções por Campylobacter , Doenças dos Bovinos , Animais , Teorema de Bayes , Infecções por Campylobacter/diagnóstico , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/veterinária , Campylobacter fetus/genética , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/epidemiologia , Testes Diagnósticos de Rotina , Genitália , Análise de Classes Latentes , Masculino , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Sensibilidade e Especificidade , UruguaiRESUMO
Infection is a rare complication of abdominal aortic aneurysms. Campylobacter fetus has special tropism for vascular endothelium. It has been reported as a cause of infected abdominal aortic aneurysms. The mortality of these patients is high, so an early recognition with a start of antibiotic therapy is crucial. In addition to medical treatment surgery may be required, which has high mortality in patients with many diseases and unstable. We report the case of an old man with infected abdominal aortic aneurysm with C. fetus, whom was decided to be treated with imipenem, due to his cardiovascular history and his general condition at admission. The patient showed a good clinical response without recurrence of symptoms. We also carry out a review of the reported cases.
Assuntos
Aneurisma Infectado , Aneurisma da Aorta Abdominal , Infecções por Campylobacter , Aneurisma Infectado/diagnóstico por imagem , Antibacterianos/uso terapêutico , Infecções por Campylobacter/diagnóstico , Infecções por Campylobacter/tratamento farmacológico , Campylobacter fetus , Humanos , MasculinoRESUMO
Resumen La infección es una complicación infrecuente de los aneurismas de la aorta abdominal. Campylobacter fetus tiene un especial tropismo por el endotelio vascular y ha sido reportado como causa de infección de aneurismas aórticos. Este tipo de infección es de alta mortalidad por lo que el reconocimiento temprano con el inicio precoz de terapia antibacteriana efectiva es clave. Además del tratamiento médico, puede requerirse cirugía, la que tiene una alta letalidad en pacientes inestables y con comorbilidades. Comunicamos el caso clínico de un adulto mayor con un aneurisma de aorta abdominal infectado por C. fetus. Dado su compromiso del estado general y antecedentes cardiovasculares se decidió tratamiento médico con imipenem, con una buena respuesta clínica y microbiológica, sin recurrencia de los síntomas. También se presenta una revisión de los casos publicados.
Abstract Infection is a rare complication of abdominal aortic aneurysms. Campylobacter fetus has special tropism for vascular endothelium. It has been reported as a cause of infected abdominal aortic aneurysms. The mortality of these patients is high, so an early recognition with a start of antibiotic therapy is crucial. In addition to medical treatment surgery may be required, which has high mortality in patients with many diseases and unstable. We report the case of an old man with infected abdominal aortic aneurysm with C. fetus, whom was decided to be treated with imipenem, due to his cardiovascular history and his general condition at admission. The patient showed a good clinical response without recurrence of symptoms. We also carry out a review of the reported cases.