RESUMEN

Consumption of raw, undercooked or contaminated animal food products is a frequent cause of Campylobacter jejuni infection. Brazil is the world's third largest producer and a major exporter of chicken meat, yet population-level genomic investigations of C. jejuni in the country remain scarce. Analysis of 221 C. jejuni genomes from Brazil shows that the overall core and accessory genomic features of C. jejuni are influenced by the identity of the human or animal source. Of the 60 sequence types detected, ST353 is the most prevalent and consists of samples from chicken and human sources. Notably, we identified the presence of diverse bla genes from the OXA-61 and OXA-184 families that confer beta-lactam resistance as well as the operon cmeABCR related to multidrug efflux pump, which contributes to resistance against tetracyclines, macrolides and quinolones. Based on limited data, we estimated the most recent common ancestor of ST353 to the late 1500s, coinciding with the time the Portuguese first arrived in Brazil and introduced domesticated chickens into the country. We identified at least two instances of ancestral chicken-to-human infections in ST353. The evolution of C. jejuni in Brazil was driven by the confluence of clinically relevant genetic elements, multi-host adaptation and clonal population growth that coincided with major socio-economic changes in poultry farming.

Asunto(s)

RESUMEN

Salmonella Enteritidis, Escherichia coli, and Campylobacter jejuni are among the most common foodborne pathogens worldwide, and poultry products are strongly associated with foodborne pathogen outbreaks. These pathogens are capable of producing biofilms on several surfaces used in the food processing industry, including polyethylene and stainless steel. However, studies on multi-species biofilms are rare. Therefore, this study aimed to develop predictive mathematical models to simulate the adhesion and removal of multispecies biofilms. All combinations of microorganisms resulted in biofilm formation with differences in bacterial counts. E. coli showed the greatest ability to adhere to both surfaces, followed by S. Enteritidis and C. jejuni. The incubation time and temperature did not influence adhesion. Biofilm removal was effective with citric acid and benzalkonium chloride but not with rhamnolipid. Among the generated models, 46 presented a significant coefficient of determination (R2), with the highest R2 being 0.88. These results provide support for the poultry industry in creating biofilm control and eradication programs to avoid the risk of contamination of poultry meat.

RESUMEN

Campylobacteriosis is currently recognized as one of the major causes of foodborne bacterial diseases worldwide. In Brazil, there is insufficient data to estimate the impact of Campylobacter in public health. The aim of this present study was to characterize a C. jejuni CJ-HBSJRP strain isolated from a hospitalized patient in Brazil by its ability to invade human Caco-2 epithelial cells, to survive in U937 human macrophages, and to assess its phenotypic antimicrobial resistance profile. In addition, prophages, virulence and antimicrobial resistance genes were search using whole-genome sequencing data. The genetic relatedness was evaluated by MLST and cgMLST analysis by comparison with 29 other C. jejuni genomes isolated from several countries. The CJ-HBSJRP strain showed an invasion percentage of 50% in Caco-2 polarized cells, 37.5% of survivability in U937 cells and was phenotypically resistant to ampicillin, ciprofloxacin and nalidixic acid. A total of 94 virulence genes related to adherence, biofilm, chemotaxis, immune modulation, invasion process, metabolism, motility and toxin were detected. The resistance genes blaOXA-605 (blaOXA-61), cmeB and mutations in the QRDR region of gyrA were also found and none prophages were detected. The MLST analysis showed 23 different STs among the strains studied. Regarding cgMLST analysis, the CJ-HBSJRP strain was genetically distinct and did not group closely to any other isolate. The results obtained reinforce the pathogenic potential of the CJHBSJRP strain and highlighted the need for more careful attention to Campylobacter spp. infections in Brazil since this pathogen has been the most commonly reported zoonosis in several countries worldwide.

Asunto(s)

RESUMEN

Campylobacter is one of the most common causes of foodborne gastroenteritis. The objective of this study was to estimate the prevalence and risk factors associated with Campylobacter jejuni and Campylobacter coli species in fresh chicken carcasses for human consumption from farmers' markets and small food stores in seven localities of Bogotá, Colombia. Ninety-one samples of fresh chicken carcasses were collected from farmers' markets and small food stores at seven localities in Bogotá. Samples were tested for Campylobacter using the real-time polymerase chain reaction (real time PCR) and isolation by plating. To analyze possible risk factors associated with Campylobacter spp. contamination in retail chicken carcasses, information was collected using a structured questionnaire and a univariate logistic regression analysis (α = 0.05) was used. Forty-two positive samples were obtained for Campylobacter spp., given a prevalence of 46.2%, of which 54.8% were to C. jejuni, 9.52% to C. coli and 35.7% to joint contaminations. C. jejuni was the most prevalent species. Risk factors found included poor cleanliness, in frequency of disinfection, type of establishment, and direct contact of chickens with other food. This study is the first report in the country on the prevalence and risk factors of Campylobacter in retail chicken.

RESUMEN

El síndrome de Guillain Barré es una enfermedad derivada del compromiso en las neuronas del sistema nervioso periférico por una respuesta descontrolada del sistema inmune que conduce daño axonal y/o desmielinización. El objetivo de este reporte fue describir los 10 primeros casos sospechosos de Síndrome de Guillain Barré en Piura. Se logró identificar la presencia de Campylobacter jejuni en las muestras de heces del 80% de los pacientes reportados. Es muy importante reconocer rápida y oportunamente al paciente con diagnóstico sospechoso de Guillain Barré, y realizar los estudios necesarios en un brote para identificar los agentes desencadenantes del cuadro.

Guillain Barré syndrome is a disease derived from compromise in neurons of the peripheral nervous system by an uncontrolled response from the immune system that leads to axonal damage and/or demyelination. The objective of this report was to describe the first 10 suspected cases of Guillain Barre Syndrome in Piura. It was possible to identify the presence of Campylobacter jejuni in the stool samples of 80% of the reported patients. It is very important to quickly and opportunely recognize the patient with a suspected diagnosis of Guillain Barré, and to carry out the necessary studies in an outbreak to identify the triggering agents of the condition.

RESUMEN

This work describes the preparation, characterization and antimicrobial activity of four palladium(II) complexes, namely, [Pd(meg)(1,10-phen)] 1, [Pd(meg)(PPh3)2] 2, [Pd(og)(1,10-phen)] 3 and [Pd(og)(PPh3)2] 4, where meg = methyl gallate, og = octyl gallate, 1,10-phen = 1,10-phenanthroline and PPh3 = triphenylphosphine. As to the chemical structures, spectral and physicochemical studies of 1-4 indicated that methyl or octyl gallate coordinates a palladium(II) ion through two oxygen atoms upon deprotonation. A chelating bidentate phenanthroline or two triphenylphosphine molecules complete the coordination sphere of palladium(II) ion, depending on the complex. The metal complexes were tested against the Mycobacterium tuberculosis H37Rv strain and 2 exhibited high activity (MIC = 3.28 µg/mL). As to the tests with Campylobacter jejuni, complex 1 showed a significant effect in reducing bacterial population (greater than 7 log CFU) in planktonic forms, as well as in the biomass intensity (IBF: 0.87) when compared to peracetic acid (IBF: 1.11) at a concentration of 400 µg/mL. The effect provided by these complexes has specificity according to the target microorganism and represent a promising alternative for the control of microorganisms of public health importance.

Asunto(s)

RESUMEN

In this study a data dependent acquisition label-free based proteomics approach was used to identify pH-dependent proteins that respond in a growth phase independent manner in Campylobacter jejuni reference strain NCTC 11168. NCTC 11168 was grown within its pH physiological normal growth range (pH 5.8, 7.0 and 8.0, µ = âˆ¼0.5 h-1) and exposed to pH 4.0 shock for 2 h. It was discovered that gluconate 2-dehydrogenase GdhAB, NssR-regulated globins Cgb and Ctb, cupin domain protein Cj0761, cytochrome c protein CccC (Cj0037c), and phosphate-binding transporter protein PstB all show acidic pH dependent abundance increases but are not activated by sub-lethal acid shock. Glutamate synthase (GLtBD) and the MfrABC and NapAGL respiratory complexes were induced in cells grown at pH 8.0. The response to pH stress by C. jejuni is to bolster microaerobic respiration and at pH 8.0 this is assisted by accumulation of glutamate the conversion of which could bolster fumarate respiration. The pH dependent proteins linked to growth in C. jejuni NCTC 11168 aids cellular energy conservation maximising growth rate and thus competitiveness and fitness.

Asunto(s)

RESUMEN

This work describes the synthesis and characterization of two metal complexes of the type [M(L1)2(phen)], where M = Pt2+ (complex I) or Pd2+ (complex II), L1 = 5-amino-1,3,4-thiadiazole-2(3H)-thiolate and phen = 1,10-phenanthroline. The in vitro antibacterial activity of these complexes was investigated in isolation and synergistically with ciprofloxacin (CIP) and erythromycin (ERY) in three strains of Campylobacter jejuni (MIC = 32 mg/L for CIP and ERY), selected from a bank of 235 strains representative of three poultry exporting states of the country (A, B and C), previously analyzed for epidemiology and resistance to CIP and ERY. A total of 53/235 (22.55%) strains showed co-resistance to CIP and ERY. Isolated resistance to CIP was higher than to ERY. Epidemiological analysis showed that resistance to CIP was more evident in state B (p < 0.0001), as well as a higher susceptibility to ERY in state C (p = 0.0028). Co-resistance was expressive in state A and in the spring and fall seasons. The evaluation of I alone and in synergy with CIP and ERY found values up to 0.25 mg/L not significant for ERY. Complex II did not show an antimicrobial effect on the three strains of tested C. jejuni. The effect provided by complex I represents a promising alternative for control of resistant strains of C. jejuni.

RESUMEN

INTRODUCTION: Campylobacter jejuni is one of the most common bacterial causes of human gastroenteritis. Despite its public health importance, the virulence factors and mechanisms underlying C. jejuni pathogenesis remain poorly understood and the relationships between these genes and the sources of the strains are not clear. We aimed to determine the virulence profiles of C. jejuni isolated from poultry and human cases of Campylobacteriosis. METHODOLOGY: A total of 50 strains of C. jejuni isolated from poultry and human cases of Campylobacteriosis were screened by polymerase chain reaction (PCR) for the presence of six pathogenic genes (flaA, iam, wlaN, cdtA, cdtB, cdtC). RESULTS: A total of 40% (10/25) of the human isolates presented only one virulence marker. In contrast, 64% (16/25) of the poultry-derived strains showed four or five virulence markers. cdtA and flaA occurred more frequently in poultry-derived strains than in human strains. Ten different virulence profiles were observed among the human isolates and 11 among the poultry strains. Only four profiles were common to both sources: profiles 3 (flaA, cdtA, cdtB, and cdtC), 5 (cdtA and cdtB), 7 (flaA and cdtB), and 10 (iam, flaA, cdtA, cdtB, and cdtC). The human-derived strains had a higher Shannon diversity index (1.9396) and Simpson index (0.8367), indicating a more diversified population than found in poultry (1.7742 and 0.7333, respectively). CONCLUSIONS: We found variations in the genetic profiles of the circulating strains based on the isolation source and genes that are potentially pathogenic to humans were detected in poultry-derived strains.

Asunto(s)

RESUMEN

Campylobacter jejuni infection is considered the most frequent factor associated with Guillain-Barré syndrome (GBS). In 2019, a large outbreak of GBS was detected in Peru, being associated with C. jejuni detected in stool samples from these patients. The aim of this study was to determine the molecular epidemiology of C. jejuni strains (ST-2993) associated with a large GBS outbreak in Peru. In this study, 26 C. jejuni strains belonging to the ST-2293, obtained from 2019 to 2020, were sequenced using Illumina technology. Five low-quality sequences were removed using bioinformatics, and 21 genomes (17 clinical strains and 4 chicken strains) were considered in the phylogenetic analysis and comparative genomics. Phylogenetic reconstruction, including genomes from international databases, showed a connection between Peruvian and Chinese GBS strains, both of them having lipooligosaccharides (LOS) locus genes related to molecular mimicry with gangliosides in peripheral nerves. Also, ST-2993 was detected in Amazon strains recovered many years before the 2019 outbreak, but with no epidemiological connection with GBS. Besides, a close relationship between human and chicken C. jejuni strains indicated chicken as one of the probable reservoirs. Finally, comparative genomics revealed differences between Chinese and Peruvian strains, including the presence of a prophage inserted into the genome. In conclusion, C. jejuni ST-2993 strains recovered from the GBS outbreak are closely related to Peruvian Amazon strains. Moreover, ST-2993 has been circulated in Peru since 2003 in the Peruvian Amazonia, showing the necessity to reinforce the epidemiological surveillance of C. jejuni to improve the prevention and control of future GBS outbreaks. IMPORTANCE This article describes the molecular epidemiology of C. jejuni strains (ST-2993) associated with a large Guillain-Barré Syndrome (GBS) outbreak in Peru, sequencing several strains recovered from GBS patients and chickens from 2019 to 2020. Phylogenetic analysis showed a connection between Peruvian and Chinese GBS strains, both of them having lipooligosaccharides (LOS) locus genes related to molecular mimicry with gangliosides in peripheral nerves. Also, ST-2993 strains were detected in isolates recovered many years before the 2019 outbreak, but with no epidemiological connection with GBS. Besides, a close relationship between human and chicken strains indicated those animals as a probable reservoir. This information will help to understand the real situation of GBS in Peru and its causal agent, C. jejuni ST-2993, showing the necessity to increase epidemiological tracking of these kinds of pathogens to detect them and avoid GBS outbreaks in the future.

Asunto(s)

RESUMEN

A wide variety of microorganism species have been used as probiotics to improve the intestinal microbial balance, control and prevent the colonization of pathogenic bacteria and promote growth in animal production. Based on the various beneficial factors of probiotic agents, this study aimed to evaluate the effectiveness of three different formulations of probiotics strains through the analysis of antagonistic capacity against common pathogens from chickens gastrointestinal tract. Three formulations composed by Bacillus, Lactobacillus, Saccharomyces, Enterococcus, Bifidobacterium and Pediococcus were tested in vitro by the method of probiotic culture spot in plates seeded with inocula of Escherichia coli, Salmonella Heidelberg and Campylobacter jejuni. The inhibition halos were measured and classified according to the degree of pathogenic bacteria inhibition. The formulation containing an association among Bacillus, Lactobacillus and Saccharomyces presented better results when compared to the other formulations.

Asunto(s)

RESUMEN

Biofilm formation has been suggested to play a significant role in the survival of pathogens in food production. Interest in evaluating alternative products of natural origin for disinfectant use has increased. However, there is a lack of information regarding the effects of biosurfactants and organic acids on Salmonella enterica serotype Enteritidis, Escherichia coli, and Campylobacter jejuni biofilms, mainly considering temperatures found in environments of poultry processing, as well as simulating the contact times used for disinfection. The aim of this study was to evaluate the antibiofilm activity of rhamnolipid, malic acid, and citric acid on the adhesion of S. Enteritidis, E. coli, and C. jejuni on polystyrene surfaces at different temperatures (4, 12, and 25 °C), compound concentrations, and times of contact (5 and 10 min), and to analyze the potential use of these compounds to disrupt formed biofilms. All three compounds exhibited antibiofilm activity under all analyzed conditions, both in the prevention and removal of formed biofilms. Contact time was less important than temperature and concentration. The antibiofilm activity of the compounds also varied according to the pathogens involved. In the food industry, compound selection must consider the temperature found in each stage of product processing and the target pathogens to be controlled.

Asunto(s)

RESUMEN

In December 2019, four children of the same school were hospitalized due to severe diarrhea, vomiting, and fever, and the mother of one child was diagnosed with hemolytic uremic syndrome (HUS). Escherichia coli O157 NM was isolated from the stool of one child, whereas Campylobacter jejuni isolates were found in feces, raw foods, environmental samples, and tap water. In addition, the same pulsed-field gel electrophoresis profile was identified in C. jejuni isolated from feces and tap water. One child died of renal failure and another due to respiratory failure. Based on symptoms and bacterial isolation, the deaths were assigned to E. coli O157 NM, but coinfection with C. jejuni may have contributed to the severity of symptoms. These were the first deaths assigned to E. coli O157 NM registered in Brazil.

Asunto(s)

RESUMEN

Annona cherimola Mill. is a native species of Ecuador cultivated worldwide for the flavor and properties of its fruit. In this study, hydrodistillation was used to isolate essential oil (EO) of fresh Annona cherimola leaves collected in Ecuadorian Sierra. The EO chemical composition was determined using a non-polar and a polar chromatographic column and enantiomeric distribution with an enantioselective column. The qualitative analysis was carried out by gas chromatography coupled to a mass spectrometer and quantitative analysis using gas chromatography equipped with a flame ionization detector. The antibacterial potency was assessed against seven Gram-negative bacteria and one Gram-positive bacterium. ABTS and DPPH assays were used to evaluate the radical scavenging properties of the EO. Spectrophotometric method was used to measure acetylcholinesterase inhibitory activity. GC-MS analysis allowed us to identify more than 99% of the EO chemical composition. Out of the fifty-three compounds identified, the main were germacrene D (28.77 ± 3.80%), sabinene (3, 9.05 ± 1.69%), ß-pinene (4, 7.93 ± 0.685), (E)-caryophyllene (10.52 ± 1.64%) and bicyclogermacrene (11.12 ± 1.39%). Enantioselective analysis showed the existence of four pairs of enantiomers, the (-)-ß-Pinene (1S, 5S) was found pure (100%). Chirimoya essential oil exhibited a strong antioxidant activity and a very strong anticholinesterase potential with an IC50 value of 41.51 ± 1.02 µg/mL. Additionally, EO presented a moderate activity against Campylobacter jejuni and Klebsiella pneumoniae with a MIC value of 500 µg/mL.

RESUMEN

This research was conducted to investigate if the administration of the probiotic Lactobacillus fermentum could influence body weight, intestinal morphometry and the cecal cytokine response in Campylobacter jejuni-infected chickens. Seventy-two 1-day old COBB 500 male chicks were allocated randomly into four experimental groups. (I) Control group (C), in which chicks were left untreated. (II) LB group, treated with L. fermentum. (III) Cj group, infected with C. jejuni and (IV) coexposure group in which both bacteria were administered. Body weight was registered and then all birds were slaughtered; samples from the small intestine and caecum were collected at 4- and 7-days post infection. The experiment lasted eleven days. Villi height and crypt depth ratios of the duodenum, jejunum and ileum were evaluated using appropriate software, while reverse transcription quantitative PCR (RT-qPCR) was utilized for assessing transcript levels of key cecal inflammatory cytokines (IL-1ß, IL-18, IL-17, IL-15, IL13 and IL-4). Campylobacter-infected birds showed lower body weight values than those supplemented with the probiotic; these birds, in turn, proved to be heavier than those reared under control conditions. L. fermentum administration improved morphometrical parameters of the duodenum, jejunum and ileum; in general, villi were larger and crypts deeper than those identified in control conditions. Moreover, the negative effects elicited by C. jejuni were not observed in chickens exposed to the probiotic. Significant differences were also determined with regards to transcript abundance of all evaluated cytokines in the caecum. C. jejuni induced a downregulation of the studied interleukins; however, such a response was heightened by administration of L. fermentum, with an increase rate of transcription that promoted a more effective response to a C. jejuni infection. The effects of experimental treatments proved to vary between sampling points. Conclusively, these results demonstrate that L. fermentum lessens the negative effects elicited by C. jejuni on body weight by alleviating the impact on intestinal morphometry and cecal cytokine response, which ultimately improve chicken growth performance.

RESUMEN

Aims: The objectives of this work were to use whole genome sequencing (WGS) to determine the antimicrobial resistance genotypes of 116 Campylobacter jejuni strains isolated in Brazil and to compare it with the results obtained by antimicrobial susceptibility testing (AST). In addition, WGS was used to uncover the phylogenetic relationship among those strains. Results: By AST, the C. jejuni strains resistant to ciprofloxacin, tetracycline, doxycycline, and erythromycin were 51 (44%), 41 (35.3%), 41 (35.3%), and 6 (5.2%), respectively. By WGS, the genes aph(3')III, aadE, blaOXA-449, blaOXA-184, blaOXA-61, and tet(O) were detected in 6 (5.2%), 3 (2.6%), 1 (0.9%), 10 (8.6%), 55 (47.4%), and 44 (38%) strains, respectively. Fifty-four (46.6%) strains showed the mutation T86I in the gyrA gene, and four (3.4%) strains presented the mutation A2075G in the 23S rRNA gene. The correlation between AST and WGS was 100% for ciprofloxacin, 97.5% for tetracyclines, and 66.7% for erythromycin. The whole genome single nucleotide polymorphism (SNP) tree clustered the C. jejuni strains into two clades comprising strains that were highly related from different sources, places, and years. Conclusion: The high rates of C. jejuni strains resistant to ciprofloxacin and tetracyclines are of concern and may represent a public health problem. WGS has a potential to be a powerful tool for the prediction of resistance of antibiotics used to treat campylobacteriosis. The results obtained by whole genome SNP analysis suggested the potential for transmission between clinical and nonclinical sources and between human and animal sources over the course of 20 years in Brazil.

Asunto(s)

RESUMEN

Campylobacter jejuni is an important foodborne pathogen with global distribution. We describe a genotyping study of a collection of C. jejuni (n=137) isolated from different broiler farms and from multiple sites along the processing line in a slaughterhouse in Argentina during 2011, 2012 and 2015. The isolates were genotyped using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Based on the PFGE results, the isolates were grouped into 26 pulsotypes. Subsequently, the isolates representing these 26 pulsotypes were chosen for MLST genotyping, which identified 16 different sequence types (STs) and 6 clonal complexes (CCs) (21, 45, 48, 353, 354, 446). Several of the STs (n=7) have not been previously reported in the PubMLST.org database. The most prevalent CCs were 21, 45 (both associated with human campylobacteriosis worldwide) and 353. This study showed high genetic diversity among C. jejuni in the broiler production environment in Argentina with novel MLST genotypes.

Asunto(s)

RESUMEN

RESUMEN Objetivo: Describir los resultados de los exámenes de laboratorio realizados en muestras biológicas de pacientes con síndrome de Guillain-Barré (SGB), recibidas en el Instituto Nacional de Salud (INS) entre los años 2018 y 2019. Materiales y métodos: Se realizó un estudio observacional en pacientes con SGB notificados en el sistema de vigilancia epidemiológica. Se obtuvieron muestras biológicas analizadas en el INS para investigar arbovirus, virus respiratorios, enterovirus y enterobacterias, entre otros. Resultados: Se recibió un total de 2051 especímenes clínicos de 906 pacientes con SGB. Tres pacientes dieron positivo al dengue y tres pacientes al Zika. En 19 pacientes, el cultivo en heces fue positivo para Campylobacter jejuni. El análisis filogenético de diez cepas de Campylobacter jejuni las clasificó como genotipo ST2993, reportado previamente en China y asociado a un brote de SGB. En 2018, hubo 12 muestras que habían dado positivo al PCR para enterovirus en el líquido cefalorraquídeo, pero ninguna pudo corroborarse con el cultivo respectivo ni con secuenciamiento de genoma completo. Un paciente dio positivo por virus de la influenza A, dos por virus de la influenza B, dos por adenovirus, cinco por virus respiratorio sincicial, y diez por rinovirus. Conclusión: Se han encontrado diversos agentes patógenos en especímenes de pacientes con SGB, sin embargo, la presencia de Campylobacter jejuni genotipo ST2993, un patógeno relacionado a brotes de SGB en varios continentes, sería el probable agente causal. Es necesario confirmar esta hipótesis con estudios analíticos y determinar la cadena de transmisión de este agente para implementar las medidas de prevención y control.

ABSTRACT Objective: To describe the results of laboratory tests performed on biological samples from patients with Guillain-Barré syndrome (GBS) submitted to the Instituto Nacional de Salud (INS) between 2018 and 2019. Materials and methods: We conducted an observational study on patients with GBS, by using data from the epidemiological surveillance system. Biological samples, previously analyzed at the INS, were obtained to study arboviruses, respiratory viruses, enteroviruses and enterobacteria, among others. Results: A total of 2,051 specimens were obtained from 906 patients with GBS. Three patients tested positive for dengue and three for Zika. In 19 patients, the stool culture was positive for Campylobacter jejuni. Phylogenetic analysis of 10 Campylobacter jejuni strains classified them as genotype ST2993, which was previously reported in China and associated to a GBS outbreak. Twelve cerebrospinal fluid samples tested positive for enterovirus by PCR in 2018, but none could be verified by culture or complete genome sequencing during the study. One patient was positive for influenza A, two for influenza B, two for adenovirus, five for respiratory syncytial virus, and ten for rinovirus. Conclusion: Several pathogens were found in samples from patients with GBS. However, we found that the genotype ST2993 of Campylobacter jejuni was the most likely causal agent, a pathogen that is related to GBS outbreaks in different continents. It is necessary to confirm this hypothesis with additional analytical studies and it is important to describe the transmission mechanism of C. jejuni genotype ST2993 in order to implement prevention and control measures.

Asunto(s)

RESUMEN

Campylobacter spp. is a bacterial agent that causes gastroenteritis in humans and may trigger Guillain-Barré Syndrome (GBS) and is also considered one of the main foodborne diseases in developed countries. Poultry and pigs are considered reservoirs of these microorganisms, as well as raw or undercooked by-products are often incriminated as a source of human infection. Treatment in human cases is with macrolide, such erythromycin, that inhibits the protein synthesis of the microorganism. This study aimed to isolate Campylobacter jejuni and Campylobacter coli from intestinal content samples of broiler chickens (n=20) and swine (n=30) to characterize the erythromycin resistance profile of the strains and to detect molecular mechanisms involved in this resistance. The minimum inhibitory concentration was determined by agar dilution. The Mismatch Amplification Mutation Assay-Polymerase Chain Reaction (MAMA-PCR) was performed to detect mutations at positions 2074 and 2075 of 23S rRNA region, in addition to PCR test to detect the erm(B) gene. From the intestinal content of broiler chickens, 18 strains of C. jejuni and two strains of C. coli were isolated, whereas, from swine samples, no C. jejuni strain and 14 strains of C. coli were isolated. All C. coli strains were resistant, and three C. jejuni strains from broilers chickens were characterized with intermediate resistance to erythromycin. The MIC of the strains ranged from ≤0.5mg/μL to ≥128mg/μL. All resistant strains had the A2075G mutation, and one strain with intermediate resistance had the A2075G mutation. However, the A2074C mutation and the erm(B) gene were not detected. High resistance levels were detected in C. coli strains isolated from swine. The MAMA-PCR is a practical tool for detecting the erythromycin resistance in Campylobacter strains.(AU)

Campylobacter spp. é um agente bacteriano causador de gastroenterite em humanos e associado à síndrome de Guillain-Barré, sendo a campilobacteriose considerada uma das principais enfermidades de origem alimentar. Aves e suínos são importantes reservatórios desses microrganismos e seus produtos derivados crus ou mal cozidos são muitas vezes incriminados como fonte de infecção humana. A primeira escolha para o tratamento em casos humanos são os antimicrobianos da classe dos macrolídeos como à eritromicina. Dentro desse contexto, o objetivo deste estudo foi isolar Campylobacter jejuni e C. coli a partir de 20 amostras de conteúdo intestinal de frangos de corte e de 30 de suínos ao abate e investigar a resistência à eritromicina das estirpes obtidas e os possíveis mecanismos moleculares envolvidos nesta resistência. A concentração inibitória mínima foi determinada pela diluição em ágar e a técnica MAMA-PCR foi utilizada para detecção de mutações nas posições 2074 e 2075 da região 23s rRNA, foi pesquisado também a presença do gene erm(B) pela PCR. A partir do conteúdo intestinal de frangos de corte foram isoladas 18 estirpes de C. jejuni e duas de C. coli, enquanto de suínos foram obtidas 14 estirpes de C. coli e nenhuma estirpe de C. jejuni. Todas as estirpes de C. coli de suínos foram identificadas como resistentes e três estirpes de C. jejuni de frangos foram caracterizadas com resistência intermediária. A CIM das estirpes variou de ≤0,5mg/μL a ≥128mg/μL. Todas as estirpes resistentes tinham a mutação A2075G e uma cepa com resistência intermediária também apresentou a mutação A2075G. Não foi detectada a mutação A2074C ou a presença do gene erm(B) em nenhuma das estirpes obtidas. Os resultados revelam um alto nível de resistência em estirpes de C. coli isoladas de suínos frente a eritromicina. A técnica MAMA PCR utilizada se constitui em uma ferramenta prática para detecção da resistência à eritromicina em estirpes de C. jejuni e C. coli.(AU)

Asunto(s)

RESUMEN

Campylobacter spp. is a bacterial agent that causes gastroenteritis in humans and may trigger Guillain-Barré Syndrome (GBS) and is also considered one of the main foodborne diseases in developed countries. Poultry and pigs are considered reservoirs of these microorganisms, as well as raw or undercooked by-products are often incriminated as a source of human infection. Treatment in human cases is with macrolide, such erythromycin, that inhibits the protein synthesis of the microorganism. This study aimed to isolate Campylobacter jejuni and Campylobacter coli from intestinal content samples of broiler chickens (n=20) and swine (n=30) to characterize the erythromycin resistance profile of the strains and to detect molecular mechanisms involved in this resistance. The minimum inhibitory concentration was determined by agar dilution. The Mismatch Amplification Mutation Assay-Polymerase Chain Reaction (MAMA-PCR) was performed to detect mutations at positions 2074 and 2075 of 23S rRNA region, in addition to PCR test to detect the erm(B) gene. From the intestinal content of broiler chickens, 18 strains of C. jejuni and two strains of C. coli were isolated, whereas, from swine samples, no C. jejuni strain and 14 strains of C. coli were isolated. All C. coli strains were resistant, and three C. jejuni strains from broilers chickens were characterized with intermediate resistance to erythromycin. The MIC of the strains ranged from ≤0.5mg/μL to ≥128mg/μL. All resistant strains had the A2075G mutation, and one strain with intermediate resistance had the A2075G mutation. However, the A2074C mutation and the erm(B) gene were not detected. High resistance levels were detected in C. coli strains isolated from swine. The MAMA-PCR is a practical tool for detecting the erythromycin resistance in Campylobacter strains.(AU)

Campylobacter spp. é um agente bacteriano causador de gastroenterite em humanos e associado à síndrome de Guillain-Barré, sendo a campilobacteriose considerada uma das principais enfermidades de origem alimentar. Aves e suínos são importantes reservatórios desses microrganismos e seus produtos derivados crus ou mal cozidos são muitas vezes incriminados como fonte de infecção humana. A primeira escolha para o tratamento em casos humanos são os antimicrobianos da classe dos macrolídeos como à eritromicina. Dentro desse contexto, o objetivo deste estudo foi isolar Campylobacter jejuni e C. coli a partir de 20 amostras de conteúdo intestinal de frangos de corte e de 30 de suínos ao abate e investigar a resistência à eritromicina das estirpes obtidas e os possíveis mecanismos moleculares envolvidos nesta resistência. A concentração inibitória mínima foi determinada pela diluição em ágar e a técnica MAMA-PCR foi utilizada para detecção de mutações nas posições 2074 e 2075 da região 23s rRNA, foi pesquisado também a presença do gene erm(B) pela PCR. A partir do conteúdo intestinal de frangos de corte foram isoladas 18 estirpes de C. jejuni e duas de C. coli, enquanto de suínos foram obtidas 14 estirpes de C. coli e nenhuma estirpe de C. jejuni. Todas as estirpes de C. coli de suínos foram identificadas como resistentes e três estirpes de C. jejuni de frangos foram caracterizadas com resistência intermediária. A CIM das estirpes variou de ≤0,5mg/μL a ≥128mg/μL. Todas as estirpes resistentes tinham a mutação A2075G e uma cepa com resistência intermediária também apresentou a mutação A2075G. Não foi detectada a mutação A2074C ou a presença do gene erm(B) em nenhuma das estirpes obtidas. Os resultados revelam um alto nível de resistência em estirpes de C. coli isoladas de suínos frente a eritromicina. A técnica MAMA PCR utilizada se constitui em uma ferramenta prática para detecção da resistência à eritromicina em estirpes de C. jejuni e C. coli.(AU)

Asunto(s)