RESUMEN
This study was designed to characterize the dynamics of infection of Mycoplasma hyopneumoniae in naïve replacement gilts after introduction to positive systems. Ninety-eight naïve gilts were monitored in three positive commercial farms (A, B, and C). The näive gilts were housed for 21 days in pens adjacently located to older gilt cohorts (named seeders), which have been naturally exposed to the positive farms. The infection dynamics was evaluated by PCR and ELISA, from laryngeal swabs and serum samples, respectively. Samples were collected at 150 (arrival), 165, 180, 210, 240, 270, 300 days of age (doa), and pre-farrowing. Infection occurred rapidly on farms A and B, taking 25.2 and 23.9 days for 95% of gilts to be PCR positive, respectively. There was no influence on the number of seeders at the time of exposure, but their absence (farm C) could explain the extended period it took for gilts to get infected (69.4 days). On average, it took 162.2 days after the first PCR detection for 85% of gilts to stop shedding the bacterium. The serology results were consistent with the herd infection curve. At pre-farrowing, 100% of gilts seroconverted and 36.7% remained PCR positive. A total of 1.33% of piglets were positive at weaning. Fifteen variants were detected among the three farms by MLVA. The acclimation protocol was efficient and easy to perform, and the presence of seeders was likely critical for early acclimation for M. hyopneumoniae.
Asunto(s)
Mycoplasma hyopneumoniae , Neumonía Porcina por Mycoplasma , Enfermedades de los Porcinos , Porcinos , Animales , Femenino , Neumonía Porcina por Mycoplasma/epidemiología , Neumonía Porcina por Mycoplasma/microbiología , Mycoplasma hyopneumoniae/genética , Granjas , Sus scrofa , Reacción en Cadena de la Polimerasa/veterinariaRESUMEN
BACKGROUND: Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) is an important zoonotic agent worldwide. The aim of this work was to compare genetically 117 S. Typhimurium isolated from different sources over 30 years in Brazil using different genomics strategies. RESULTS: The majority of the 117 S. Typhimurium strains studied were grouped into a single cluster (â 90%) by the core genome multilocus sequence typing and (â 77%) by single copy marker genes. The phylogenetic analysis based on single nucleotide polymorphism (SNP) grouped most strains from humans into a single cluster (â 93%), while the strains isolated from food and swine were alocated into three clusters. The different orthologous protein clusters found for some S. Typhimurium isolated from humans and food are involved in metabolic and regulatory processes. For 26 isolates from swine the sequence types (ST) 19 and ST1921 were the most prevalent ones, and the ST14, ST64, ST516 and ST639 were also detected. Previous results typed the 91 S. Typhimurium isolates from humans and foods as ST19, ST313, ST1921, ST3343 and ST1649. The main prophages detected were: Gifsy-2 in 79 (67.5%) and Gifsy-1 in 63 (54%) strains. All of the S. Typhimurium isolates contained the acrA, acrB, macA, macB, mdtK, emrA, emrB, emrR and tolC efflux pump genes. CONCLUSIONS: The phylogenetic trees grouped the majority of the S. Typhimurium isolates from humans into a single cluster suggesting that there is one prevalent subtype in Brazil. Regarding strains isolated from food and swine, the SNPs' results suggested the circulation of more than one subtype over 30 years in this country. The orthologous protein clusters analysis revealed unique genes in the strains studied mainly related to bacterial metabolism. S. Typhimurium strains from swine showed greater diversity of STs and prophages in comparison to strains isolated from humans and foods. The pathogenic potential of S. Typhimurium strains was corroborated by the presence of exclusive prophages of this serovar involved in its virulence. The high number of resistance genes related to efflux pumps is worrying and may lead to therapeutic failures when clinical treatment is needed.
RESUMEN
The intensification of pig production and advances in the sanitary control of herds profoundly changed the profile of risk attributed to pork consumption. In the actual scenario, most microorganisms related to macroscopic lesions observed in the post mortem inspection are not transmitted by food, while foodborne bacteria of importance to consumer health do not cause macroscopic lesions. In Brazil, the "Ministério da Agricultura, Pecuária e Abastecimento" requested a scientific opinion on the prioritizing of pathogens potentially transmitted by unprocessed pork. After conducting a qualitative risk assessment, only Salmonella enterica was classified as of high risk to consumers. The present study was part of the validation step of the risk assessment and aimed to investigate the frequency of S. enterica, Yersinia enterocolitica and Listeria monocytogenes and hygienic-sanitary indicators in pig carcasses of pigs rose under intensive production and slaughtered under the Federal Inspection System in three slaughterhouses located in Southern Brazil. Additionally, the antimicrobial resistance profile of the isolated pathogens was also investigated. A total of 378 carcasses were sampled by superficial sponges before the chilling step in three slaughterhouses. Samples were investigated for the presence of the three aforementioned pathogens and subjected to enumeration of Colony Formation Units (log CFU.cm-1) of total aerobic mesophiles (TAM) and Enterobacteriaceae. Salmonella strains were tested by disc diffusion test for resistance to eleven antimicrobials. There were significantly statistical differences (p<0.0001) on the median counts of both indicators between the slaughterhouses. The median of TAM was very close for Slaughterhouses A and B: 1.573 log CFU.cm-1 and 1.6014 log CFU.cm-1, respectively. While in Slaughterhouse C, a higher TAM median was detected (2.216 log CFU.cm-1). A similar profile was observed regarding to Enterobacteriaceae, and medians were calculated as follow: -0.426 log CFU.cm-1 in Slaughterhouse A; 0.2163 log CFU.cm-1 in B; and 0.633 log CFU.cm-1 in C. Regarding the pathogens investigated, L. monocytogenes was not detected and only one carcass from Slaughterhouse C was positive for Y. enterocolitica. Thus, the results suggest a very low prevalence of L. monocytogenes and Y. enterocolitica in the sampled population. A total of 65 (17.2%) carcasses were positive for S. enterica, with a difference in frequencies between slaughterhouses and slaughter days. The prevalence of Salmonella positive carcasses was higher in the Slaughterhouse C (25.4%; CI 95% 19-32%) in comparison with A (9.5%; CI 95% 9-14%) and B (18.3%; CI 95% 12-24%). There was no significantly statistical association between Enterobacteriaceae counts and Salmonella isolation on carcass surface (p=0.69). The slaughtering day, nested within the slaughterhouse, explains 31.3% of Salmonella prevalence variability. S. Typhimurium (38.1%) was the most prevalent, followed by S. Infantis (30.1%). Among the 61 Salmonella strains tested for resistance to antimicrobials, 18 (31.6%) were full-susceptible. No strain displayed resistance to azithromycin, ceftazidime, cefotaxime and meropenem. The highest resistance frequency was displayed to tetracycline (54.1%), followed by ampicillin (50.82%), nalidixic acid (42.62%) and chloramphenicol (42.62). Multi-resistance was detected in 52.54% of the, strains. In conclusion, S. enterica is more prevalent in pre-chill pig carcasses than Y. enterocolitica and L. monocytogenes and thus should be prioritized in monitoring and control programs at slaughter. Salmonella serovars varied among slaughterhouses and present significant differences in their resistance to antimicrobials. Slaughterhouses that present higher medians of TAM or Enterobacteriaceae in a monitoring period may have higher S. enterica prevalences as well. However, there is a high variation of S. enterica prevalence among slaughter days, which cannot be always related to the hygienic indicators counts observed on a given day.(AU)
A intensificação da produção de suínos e os avanços no controle sanitário dos rebanhos alterou de forma importante o perfil de risco do consumo de carne suína. No cenário atual, a maioria dos microrganismos causadores de lesões macroscópicas detectáveis na inspeção post mortem não são transmissíveis por alimentos, enquanto bactérias de importância como causadoras de doenças transmitidas por alimentos não causam lesões macroscópicas. No Brasil, o Ministério da Agricultura, Pecuária e Abastecimento solicitou uma opinião científica sobre a priorização de patógenos potencialmente transmitidos pela carne suína in natura. Após conduzir uma avaliação de risco qualitativa, apenas Salmonella enterica foi classificada como de alto risco para o consumidor. O presente estudo foi parte da etapa de validação da avaliação de risco e objetivou: investigar a frequência de S. enterica, Yersinia enterocolitica e Listeria. monocytogenes; e enumerar indicadores higiênico-sanitários em carcaças de suínos abatidos sob inspeção federal em frigoríficos dedicados ao abate de suínos sob sistema intensivo de criação no sul do Brasil. Além disso, o perfil de resistência a antimicrobianos dos patógenos isolados foi investigado. A superfície de um total de 378 carcaças foi amostrada por esponjas, na etapa de pré-resfriamento em três matadouros frigoríficos (A, B, C). As amostras foram investigadas quanto à presença dos três patógenos acima mencionados e quanto à enumeração de Unidades Formadoras de Colônia (log UFC.cm-1) de mesófilos aeróbios totais (MAT) e Enterobacteriaceae. As cepas isoladas de Salmonella foram testadas quanto à resistência a onze antimicrobianos pela técnica de disco difusão. As medianas de contagem de ambos os indicadores apresentaram diferença significativa (p<0,0001) entre matadouros-frigoríficos. A mediana de MAT foi bastante próxima para A e B (1,573 log UFC.cm-1 e 1,6014 log UFC.cm-1, respectivamente), enquanto em C uma mediana de MAT mais elevada foi determinada (2,216 log CFU.cm-1). Um perfil semelhante foi observado em relação a Enterobacteriaceae, sendo as medianas calculadas para A, B e C, respectivamente: -0,426 log CFU.cm-1; 0,2163 log UFC.cm-1; e 0,633 log UFC.cm-1. Em relação aos patógenos investigados, L. monocytogenes não foi detectada e apenas uma carcaça, do Matadouro C, foi positiva para Y. enterocolitica. Portanto, os resultados sugerem uma prevalência muito baixa desses patógenos na população amostrada. Em um total de 65 (17,2%) carcaças houve isolamento de S. enterica, com diferença nas frequências observadas entre matadouros e dias de abate. A prevalência de carcaças positivas para S. enterica foi maior no Matadouro C (25,4%; IC95% 19-32%) em comparação com A (9,5%; IC95% 9-14%) e B (18,3%; IC95% 12-24%). Não houve associação estatística entre o número de Enterobacteriaceae e o isolamento de S. enterica na superfície das carcaças (p=0,69). O dia de abate agrupado por frigorífico explica 31,3% da variação na prevalência de Salmonella. O sorovar mais frequente de S. enterica foi Typhimurium (38,1%) seguido de S. Infantis (30,1%). Entre as 61 cepas de S. enterica testadas quanto à resistência a antimicrobianos, 18 (31,6%) foram totalmente suscetíveis aos antimicrobianos testados. Nenhuma cepa apresentou resistência a azitromicina, ceftazidima, cefotaxima e meropenem. As maiores frequências de resistência foram demonstradas contra tetraciclina (54,1%), ampicilina (50,8%), ácido nalidíxico (42,62%) e cloranfenicol (42,62%). Em 52,54% das cepas foi detectada multi-resistência. Em conclusão, S. enterica é mais prevalente em carcaças suínas no pré-resfriamento do que Y. enterocolitica e L. monocytogenes. Portanto, S. enterica deve ser priorizada em programas de monitoramento e controle ao abate. Os sorovares de Salmonella variam entre matadouros e apresentam diferenças significativas na resistência a antimicrobianos. Matadouros de suínos que apresentam medianas de MAT e Enterobacteriaceae num período de monitoramento podem apresentar também prevalências mais de altas de presença de S. enterica. Entretanto, há uma alta variabilidade na frequência de S. enterica entre dias de abate, e nem sempre há relação entre essa frequência e a contagem de indicadores higiênico-sanitários determinados num determinado dia.(AU)
Asunto(s)
Animales , Yersinia enterocolitica/aislamiento & purificación , Salmonella enterica/aislamiento & purificación , Farmacorresistencia Bacteriana , Carne de Cerdo/microbiología , Listeria monocytogenes/aislamiento & purificación , Mataderos , Sus scrofaRESUMEN
Salmonella is a leading cause of bacterial foodborne-related illness and pork products are a food-associated source. With > 50% of U.S. swine herds testing positive for Salmonella, asymptomatic carrier pigs that shed Salmonella in their feces are a food safety and environmental contamination issue. Herd level surveillance of Salmonella shedding status is useful, but collection of feces and culture methods for Salmonella detection are laborious and time-consuming. Surveillance for Salmonella-exposure through detection of Salmonella-specific serum antibody is a reliable method, but presents labor and animal-welfare issues. Oral fluids are a reliable, antemortem sample with proven utility for surveillance in the swine industry. We tested oral fluid samples as a potential non-invasive, repeatable sample type for the presence of Salmonella-specific antibodies. An indirect enzyme-linked immunosorbent assay (ELISA) detected anti-Salmonella IgG, IgM, and predominantly IgA in oral fluids from Salmonella enterica serovar Typhimurium-exposed pigs. Furthermore, with minor modifications, a commercial ELISA-based kit also detected Salmonella-specific antibodies in oral fluids. Collectively, oral fluids may serve as a prospective surveillance tool for herd level monitoring of Salmonella exposure.
RESUMEN
The aim of the present study was to investigate the presence of Salmonella spp. in samples collected from beef meat at three points of the slaughter line (after skinning, washing and cooling) at three slaughterhouses in Brazil that export meat. Detection was based on ISO 6579:2002 and confirmed by PCR and qPCR. The isolates were typified using slide agglutination tests and PFGE. The antibiotic sensitivity profile was determined using the disk diffusion method. Contamination was detected in only one slaughterhouse. The overall frequency of contamination by Salmonella spp. was 6.7% of carcasses (6/90) and 2.6% of carcass surface samples (7/270). All isolates were confirmed by PCR and qPCR. The serological analysis and the PFGE showed a single profile: Typhimurium. The strains demonstrated 100% susceptibility to ampicillin, cefotaxime, ciprofloxacin, chloramphenicol, gentamicin and tetracycline. Positive carcasses after cooling pose a direct risk to consumers, since the meat is considered ready to be marketed after this process.(AU)
O objetivo deste trabalho foi investigar a presença de Salmonella spp. em amostras coletadas de carcaças de bovinos, em três pontos da linha de abate (após a esfola, lavagem e refrigeração) de três frigoríficos exportadores no Brasil. A detecção foi realizada pela ISO 6579:2002, e confirmada por PCR e qPCR. Os isolados foram tipificados por testes de soroaglutinação e PFGE e avaliado o perfil de sensibilidade aos antibióticos pelo método de difusão em disco. A contaminação foi detectada em apenas um abatedouro-frigorífico. As contaminações das carcaças (n=90) e amostras de carne (n=270) por Salmonella spp. foram 6 (6,7%) e 7 (2,6%), respectivamente. Todos os isolados foram confirmados por PCR e qPCR. A análise sorológica e o PFGE mostraram um único perfil: Typhimurium. As cepas apresentaram 100% de suscetibilidade à ampicilina, cefotaxima, ciprofloxacina, cloranfenicol, gentamicina e tetraciclina. As carcaças positivas após a refrigeração apresentam um risco direto para o consumidor, uma vez que, após este processo, a carne está pronta para ser comercializada.(AU)
Asunto(s)
Animales , Bovinos , Infecciones por Salmonella , Salmonella typhimurium , Farmacorresistencia Microbiana , Enfermedades de los Bovinos/microbiología , Enfermedades de los Bovinos/epidemiología , Industria de la Carne , Carne Roja/microbiología , Microbiología de Alimentos , Brasil/epidemiología , MataderosRESUMEN
MicroRNAs (miRNAs) are small, non-coding RNAs that regulate eukaryotic gene expression at the post-transcriptional level. In addition to their involvement in a variety of biological processes, miRNAs are implicated in the eukaryotic response to bacterial pathogens. The objective of this study was to identify miRNAs involved in the regulation of the porcine response to the human foodborne pathogen, Salmonella enterica serovar Typhimurium (S. Typhimurium). Differential expression analysis from the whole blood of pigs over a 7-day period following S. Typhimurium challenge identified 50 miRNAs, many of which are implicated in functional pathways associated with NF-κB signaling and apoptosis (e.g., ssc-let-7c, ssc-miR-21). Transcriptional analyses of whole blood mRNA identified the differential expression of several genes involved in NF-κB signaling and apoptosis (e.g., IL10, CBX4, TGFB2) whose mRNAs are predicted targets of miRNAs identified in our study. Overall, our data identified porcine miRNAs that are differentially expressed following S. Typhimurium challenge, thereby defining regulatory factors to target for controlling the porcine response to this human foodborne pathogen.
Asunto(s)
MicroARNs/genética , FN-kappa B/metabolismo , Salmonelosis Animal/genética , Salmonella typhimurium/patogenicidad , Enfermedades de los Porcinos/microbiología , Animales , Apoptosis , Línea Celular , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Redes Reguladoras de Genes , Transducción de Señal , Porcinos , Enfermedades de los Porcinos/genéticaRESUMEN
PURPOSE: Non-host-adapted Salmonella serovars, including the common human food-borne pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium), are opportunistic pathogens that can colonize food-producing animals without causing overt disease. Interventions against Salmonella are needed to enhance food safety, protect animal health and allow the differentiation of infected from vaccinated animals (DIVA). METHODOLOGY: An attenuated S. Typhimurium DIVA vaccine (BBS 866) was characterized for the protection of pigs following challenge with virulent S. Typhimurium. The porcine transcriptional response to BBS 866 vaccination was evaluated. RNA-Seq analysis was used to compare gene expression between BBS 866 and its parent; phenotypic assays were performed to confirm transcriptional differences observed between the strains. RESULTS: Vaccination significantly reduced fever and interferon-gamma (IFNγ) levels in swine challenged with virulent S. Typhimurium compared to mock-vaccinated pigs. Salmonella faecal shedding and gastrointestinal tissue colonization were significantly lower in vaccinated swine. RNA-Seq analysis comparing BBS 866 to its parental S. Typhimurium strain demonstrated reduced expression of the genes involved in cellular invasion and bacterial motility; decreased invasion of porcine-derived IPEC-J2 cells and swimming motility for the vaccine strain was consistent with the RNA-Seq analysis. Numerous membrane proteins were differentially expressed, which was an anticipated gene expression pattern due to the targeted deletion of several regulatory genes in the vaccine strain. RNA-Seq analysis indicated that genes involved in the porcine immune and inflammatory response were differentially regulated at 2 days post-vaccination compared to pre-vaccination. CONCLUSION: Evaluation of the S. Typhimurium DIVA vaccine indicates that vaccination will provide both swine health and food safety benefits.
Asunto(s)
Salmonelosis Animal/prevención & control , Vacunas contra la Salmonella/administración & dosificación , Salmonella typhimurium/inmunología , Enfermedades de los Porcinos/prevención & control , Vacunas de ADN/administración & dosificación , Animales , Anticuerpos Antibacterianos/sangre , Derrame de Bacterias , Línea Celular , Ensayo de Inmunoadsorción Enzimática , Heces/microbiología , Expresión Génica , Perfilación de la Expresión Génica , Humanos , Interferón gamma/inmunología , Salmonelosis Animal/inmunología , Salmonelosis Animal/microbiología , Vacunas contra la Salmonella/inmunología , Salmonella typhimurium/metabolismo , Salmonella typhimurium/patogenicidad , Porcinos , Enfermedades de los Porcinos/microbiología , Vacunación , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/inmunología , Vacunas de ADN/inmunologíaRESUMEN
Swine are often asymptomatic carriers of Salmonella spp., a leading cause of human bacterial foodborne disease. Vaccination against Salmonella is effective for protecting animal health and enhancing food safety. However, with >2500 Salmonella serovars, current vaccines for swine offer limited cross-protection against heterologous serovars. Also, existing vaccines can interfere with surveillance programs that monitor the Salmonella status of swine herds. To overcome Salmonella vaccine limitations, we rationally designed and constructed an attenuated Salmonella enterica serovar Typhimurium vaccine (BBS 866) by deleting multiple small regulatory RNA (sRNA) genes (omrA, omrB, rybB, micA, and invR) in combination with an rfaH mutation. We vaccinated swine intranasally at 3-weeks of age with PBS (mock-vaccinated), BBS 866 or BBS 202 (S. Typhimurium rfaH, Bearson et al., Front Vet Sci 2014;1:9.) and challenged at 7-weeks of age with virulent S. Choleraesuis, a swine pathogen. Vaccination with BBS 866 enhanced protection against S. Choleraesuis by significantly limiting the duration of fever, weight loss, the levels of circulating INFγ, and the total number of swine with S. Choleraesuis septicemia. Vaccination with either BBS 866 or BBS 202 significantly reduced S. Choleraesuis colonization of both systemic (spleen and liver) and gastrointestinal (Peyer's Patch, Ileocecal lymph nodes, and cecum) tissues. Similar to our earlier report for BBS 202, the BBS 866 vaccine strain can be used in swine without compromising the differentiation of infected from vaccinated animals (DIVA). Therefore, the attenuated S. Typhimurium BBS 866 strain, containing mutations in rfaH and multiple sRNAs, addresses the limitations of current Salmonella vaccines by providing cross-protection against Salmonella serovars in swine without interfering with established monitoring programs for Salmonella surveillance.
Asunto(s)
Protección Cruzada , Salmonelosis Animal/prevención & control , Vacunas contra la Salmonella/uso terapéutico , Enfermedades de los Porcinos/prevención & control , Animales , Eliminación de Gen , Interferón gamma/sangre , Mutación , Vacunas contra la Salmonella/administración & dosificación , Salmonella typhimurium/genética , Bazo/microbiología , Porcinos , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/uso terapéuticoRESUMEN
Swine are often asymptomatic carriers of Salmonella spp., and interventions are needed to limit colonization of swine to enhance food safety and reduce environmental contamination. We evaluated the attenuation and potential vaccine use in pigs of a Salmonella enterica serovar Typhimurium mutant of rfaH, the gene encoding the RfaH antiterminator that prevents premature termination of long mRNA transcripts. Pigs inoculated with wild-type S. Typhimurium exhibited a significant elevation in average body temperature (fever) at 1 and 2 days post-inoculation; rfaH-inoculated pigs did not (n = 5/group). During the 7-day trial, a significant reduction of Salmonella in the feces, tonsils, and cecum were observed in the rfaH-inoculated pigs compared to wild-type inoculated pigs. To determine whether vaccination with the rfaH mutant could provide protection against wild-type S. Typhimurium challenge, two groups of pigs (n = 14/group) were intranasally inoculated with either the rfaH mutant or a PBS placebo at 6 and 8 weeks of age and challenged with the parental, wild-type S. Typhimurium at 11 weeks of age. The average body temperature was significantly elevated in the mock-vaccinated pigs at 1 and 2 days post-challenge, but not in the rfaH-vaccinated pigs. Fecal shedding at 2 and 3 days post-challenge and colonization of intestinal tract tissues at 7 days post-challenge by wild-type S. Typhimurium was significantly reduced in the rfaH-vaccinated pigs compared to mock-vaccinated pigs. Serological analysis using the IDEXX HerdChek Swine Salmonella Test Kit indicated that vaccination with the rfaH mutant did not stimulate an immune response against LPS. These results indicate that vaccination of swine with the attenuated rfaH mutant confers protection against challenge with virulent S. Typhimurium but does not interfere with herd level monitoring for Salmonella spp., thereby allowing for differentiation of infected from vaccinated animals (DIVA).
RESUMEN
The aim of this study was to examine the Salmonella serovars and antimicrobial resistance within an animal-based agriculture river system. The study area consisted of a 1,345 ha upper part of Pinhal catchment. A total of 384 samples were collected in four years of monitoring. Salmonella was isolated from 241 samples (62.7%), resulting in 324 isolates. The highest number of Salmonella sp. occurred in samples associated with sites with high stoking density animal unit per hectare. It was possible to demonstrate the variability of serovars in the study area: 30 different serovars were found and at least 11 per monitoring site. Thirty-three potentially related isolates were genotyped by PFGE, one major clone was observed in serovar Typhimurium, which occurred in animal feces (swine and bovine), and different sites and samplings proving the cross-contamination and persistence of this specific clone. Among 180 isolates submitted to an antimicrobial susceptibility test, 50.5% were susceptible to all 21 antimicrobials tested and 54 different profiles were found. In the current study, 49.5% of the tested isolates were resistant to at least one antimicrobial, and multi-resistance occurred in 18% of isolates. Results indicate a close interaction between animal-based agriculture, Salmonella, and antimicrobial resistance.
Asunto(s)
Crianza de Animales Domésticos , Farmacorresistencia Bacteriana , Ríos/microbiología , Salmonella/crecimiento & desarrollo , Contaminación del Agua/análisis , Pruebas de Sensibilidad Microbiana , Ríos/química , Salmonella/efectos de los fármacos , Salmonella/aislamiento & purificación , Microbiología del AguaRESUMEN
Controlling Salmonella in the food chain is complicated by the ability of Salmonella to colonize livestock without causing clinical symptoms/disease. Salmonella-carrier animals are a significant reservoir for contamination of naïve animals, the environment, and our food supply. Salmonella carriage and shedding in pigs varies greatly both experimentally and on-farm. To investigate the dynamics between the porcine intestinal microbiota and Salmonella shedding, we temporally profiled the microbiota of pigs retrospectively classified as low and high Salmonella-shedders. Fifty-four piglets were collectively housed, fed and challenged with 10(9)Salmonella enterica serovar Typhimurium. Bacterial quantitation of Salmonella in swine feces was determined, and total fecal DNA was isolated for 16S rRNA gene sequencing from groups of high-shedder, low-shedder, and non-inoculated pigs (n=5/group; 15 pigs total). Analyses of bacterial community structures revealed significant differences between the microbiota of high-shedder and low-shedder pigs before inoculation and at 2 and 7 days post-inoculation (d.p.i.); microbiota differences were not detected between low-shedder and non-inoculated pigs. Because the microbiota composition prior to Salmonella challenge may influence future shedding status, the "will-be" high and low shedder phylotypes were compared, revealing higher abundance of the Ruminococcaceae family in the "will-be" low shedders. At 2d.p.i., a significant difference in evenness for the high shedder microbiota compared to the other two groups was driven by decreases in Prevotella abundance and increases in various genera (e.g. Catenibacterium, Xylanibacter). By 21 d.p.i., the microbial communities of high-shedder and low-shedder pigs were no longer significantly different from one another, but were both significantly different from non-inoculated pigs, suggesting a similar Salmonella-induced alteration in maturation of the swine intestinal microbiota regardless of shedding status. Our results correlate microbial shifts with Salmonella shedding status in pigs, further defining the complex interactions among the host, pathogen, and microbiota of this important public health issue and food safety concern.
Asunto(s)
Microbiota/fisiología , Salmonelosis Animal/microbiología , Salmonella typhi/fisiología , Animales , Derrame de Bacterias , Temperatura Corporal , ADN Bacteriano/clasificación , ADN Bacteriano/genética , ADN Bacteriano/aislamiento & purificación , Diarrea/microbiología , Heces/microbiología , Tracto Gastrointestinal/microbiología , Interacciones Huésped-Patógeno , Prevotella/crecimiento & desarrollo , Reacción en Cadena en Tiempo Real de la Polimerasa , Salmonella typhi/genética , Salmonella typhi/aislamiento & purificación , PorcinosRESUMEN
Swine can carry Salmonella strains that may be transmitted to humans by pork products. This investigation determined the distribution and types of Salmonella in 12 swine finishing herds and a slaughter facility in Santa Catarina, Brazil. A total of 1258 samples, consisting of environmental, feed, carcass, lymph node, and fecal material were collected and submitted to bacteriological isolation of Salmonella. From 487 positive samples, 1255 isolates were recovered and confirmed to be Salmonella. The distribution of positive samples was as follows: finishing pen floors 26% (16/61); feed 29% (42/143); feces 44% (52/119); pooled feces 59% (35/59); slaughter holding pens 90% (36/40); lymph nodes 46% (220/478); pre-chilled carcass surfaces 24% (24/98); and post-chilled carcass surfaces 24% (62/260). The most prevalent serovars were Typhimurium, Panama, Senftenberg, Derby, and Mbandaka. By pulsed-field gel electrophoresis, 1071 isolates were subtyped using XbaI, and duplicate isolates were removed. From the remaining 747 isolates, 163 macrorestriction profiles (pulsotypes) were identified. Six pulsotypes were considered very frequent, occurring in 33 isolates or more. The multiple correspondence analyses showed correlations between pulsotypes from shedding pigs (feces), herd environment (pen floors), and subiliac and prescapular lymph nodes and between lairage and carcass surface samples before and after chilling. All sources of Salmonella investigated contributed to the carrier state; however, pre-slaughter contamination at lairage was the variable most strongly associated with carcass contamination. A total of 59 different antimicrobial resistance profiles were observed in 572 Salmonella isolates. From these isolates, 17% (97/572) were susceptible to all 15 antibiotics tested, 83% (475/572) were resistant to at least one, and 43% (246/572) were resistant to four or more antibiotics (multi-resistant). The AmpGenKanTet profile was the most prevalent in carcass isolates and was associated with farm origin.