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1.
J Food Prot ; 76(7): 1137-44, 2013 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-23834787

RESUMEN

A pilot study was conducted to assess the transfer of Escherichia coli from animal slurry fertilizer to lettuce, with E. coli serving as an indicator of fecal contamination and as an indicator for potential bacterial enteric pathogens. Animal slurry was applied as fertilizer to three Danish agricultural fields prior to the planting of lettuce seedlings. At harvest, leaves (25 g) of 10 lettuce heads were pooled into one sample unit (n = 147). Soil samples (100 g) were collected from one field before slurry application and four times during the growth period (n = 75). E. coli was enumerated in slurry, soil, and lettuce on 3M Petrifilm Select E. coli Count Plates containing 16 mg/liter streptomycin, 16 mg/liter ampicillin, or no antimicrobial agent. Selected E. coli isolates (n = 83) originating from the slurry, soil, and lettuce were genotyped by pulsed-field gel electrophoresis (PFGE) to determine the similarity of isolates. The slurry applied to the fields contained 3.0 to 4.5 log CFU/g E. coli. E. coli was found in 36 to 54% of the lettuce samples, streptomycin-resistant E. coli was found in 10.0 to 18.0% of the lettuce samples, and ampicillin-resistant E. coli in 0 to 2.0% of the lettuce samples (the detection limit was 1 log CFU/g). The concentration of E. coli exceeded 2 log CFU/g in 19.0% of the lettuce samples. No E. coli was detected in the soil before the slurry was applied, but after, E. coli was present until the last sampling day (harvest), when 10 of 15 soil samples contained E. coli. A relatively higher frequency of E. coli in lettuce compared with the soil samples at harvest suggests environmental sources of fecal contamination, e.g., wildlife. The higher frequency was supported by the finding of 21 different PFGE types among the E. coli isolates, with only a few common PFGE types between slurry, soil, and lettuce. The frequent finding of fecal-contaminated lettuce indicates that human pathogens such as Salmonella and Campylobacter can be present and represent food safety hazards.


Asunto(s)
Escherichia coli/crecimiento & desarrollo , Contaminación de Alimentos/análisis , Lactuca/microbiología , Viabilidad Microbiana , Microbiología del Suelo , Animales , Recuento de Colonia Microbiana , Escherichia coli/aislamiento & purificación , Heces/microbiología , Microbiología de Alimentos , Humanos , Proyectos Piloto , Verduras/microbiología
2.
Vet Microbiol ; 163(3-4): 373-7, 2013 May 03.
Artículo en Inglés | MEDLINE | ID: mdl-23391438

RESUMEN

There is a need for more sensitive detection methods to improve effectiveness of control programmes of Salmonella enterica subsp. enterica serotypes (Salmonella) in cattle. We assessed the performance of a rapid, molecular-based, real-time PCR (rt-PCR) method against the conventional bacteriological culture-reference method (BCRM) on cattle faecal samples for detection of sub-clinical Salmonella infections in cattle. Thirty faecal samples were artificially contaminated with either 10 or 50 CFU of one of five strains of S. Dublin (SD) and S. Typhimurium (ST). The overall detection sensitivity of both rt-PCR and BCRM was 100% for ST and 78% for SD. Furthermore, 163 faecal samples from cattle herds with suspected Salmonella infection were tested to compare the relative performance of rt-PCR to BCRM on samples from naturally infected herds. The relative sensitivity of rt-PCR was 20% (3/15 BCRM positive samples) while the relative specificity and accuracy was 99% and 92%, respectively. Both methods had limitations for detecting low levels of SD (<1 CFU/g). Hence, the evaluated rt-PCR method did not provide a sensitive alternative to the BCRM for detection of bacteria in faecal samples of sub-clinically, Salmonella-infectious cattle.


Asunto(s)
Técnicas Bacteriológicas/veterinaria , Enfermedades de los Bovinos/diagnóstico , Heces/microbiología , Técnicas de Diagnóstico Molecular/veterinaria , Salmonelosis Animal/diagnóstico , Animales , Técnicas Bacteriológicas/normas , Bovinos , Técnicas de Diagnóstico Molecular/normas , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Salmonella/genética , Sensibilidad y Especificidad
3.
Animal ; 7(1): 66-74, 2013 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-23031645

RESUMEN

This study aimed to assess whether inclusion of chicory or lupine (prebiotics) in the diet of pre-slaughter pigs for just 1 or 2 weeks could change the composition of their intestinal microbiota, stimulate the growth of bifidobacteria and help to lower the amount of thermoplilic Campylobacter spp. (mainly Campylobacter jejuni and Campylobacter coli), which are a major cause of food-borne infections in humans. A total of 48 pigs that had an initial live weight of 90 kg were fed with either a lupine (organic concentrate with 25% blue lupine seeds), chicory (organic concentrate with 10% dried chicory roots) or control (100% organic concentrate) diet for 1 week (24 pigs) or 2 weeks (24 pigs) before slaughter. The Campylobacter spp. level in rectal faecal samples after 0, 1 and 2 weeks of feeding and in the luminal content from ileum, caecum and colon at slaughter was determined by direct plating on modified charcoal-cefoperazone-deoxycholate agar plates. DNA extracted from the luminal content of distal ileum and caecum was used for terminal restriction fragment length polymorphism (T-RFLP) analysis of the composition of intestinal microbiota and for measuring the amount of bifidobacterial and total bacterial DNA by quantitative real-time PCR (qPCR). Campylobacter spp. were excreted by all pigs and present in the luminal content from distal ileum to midway colon with particularly high numbers in the caecum, but the excretion was reduced by 10-fold in pigs fed lupines for 1 week as compared with control- and chicory-fed pigs (mean log(10) 2.9 v. 4.1 CFU/g; P < 0.05). The qPCR analysis showed that feeding with lupines resulted in higher levels of bifidobacteria in caecum as compared with the other diets (P < 0.05). T-RFLP analysis showed that four of the most abundant bacteria with terminal restriction fragment values >5% relative to the intensity of total abundance differed between the feed treatments (P < 0.05). Therefore, this study showed that even a short-term alternative feeding strategy with prebiotics in the diet of pre-slaughter pigs elicited changes in the composition of the intestinal microbiota, where lupine increased the level of bifidobacteria in caecum and reduced the Campylobacter spp. excretion level after 1 week.


Asunto(s)
Bifidobacterium/aislamiento & purificación , Campylobacter/aislamiento & purificación , Cichorium intybus/metabolismo , ADN Bacteriano/genética , Lupinus/metabolismo , Sus scrofa/microbiología , Alimentación Animal/análisis , Animales , Bifidobacterium/clasificación , Bifidobacterium/genética , Campylobacter/clasificación , Campylobacter/genética , Ciego/microbiología , Colon/microbiología , Recuento de Colonia Microbiana/veterinaria , Dieta/veterinaria , Suplementos Dietéticos/análisis , Heces/microbiología , Femenino , Íleon/microbiología , Masculino , Polimorfismo de Longitud del Fragmento de Restricción , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Sus scrofa/fisiología , Factores de Tiempo
4.
Epidemiol Infect ; 139(7): 1075-80, 2011 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-21062531

RESUMEN

The study's objectives were to determine herd- and animal-level prevalence and herd-level risk factors for Salmonella in dairy-bred veal calves at slaughter in Denmark. In total, 1296 faecal samples were collected at five cattle abattoirs in Denmark during 2007-2008. The animals came from 71 randomly selected specialized veal-calf producers that delivered more than 100 animals to slaughter per year. Salmonella Dublin bacteria were isolated from 19 samples from 12 herds and Salmonella Typhimurium was isolated from one sample. The apparent prevalence of herds delivering Salmonella-shedding animals to slaughter was 18% (95% CI 9-27). The overall estimated true prevalence of shedding calves at slaughter was 1.3%. Veal-calf herds that purchased animals from herds not classified as low risk in the Danish Salmonella surveillance programme had significantly (P=0.03) higher risk of delivering Salmonella-shedding calves to slaughter. The results emphasize the importance of efforts in the dairy industry to ensure food safety for consumers.


Asunto(s)
Enfermedades de los Bovinos/epidemiología , Salmonelosis Animal/epidemiología , Mataderos/estadística & datos numéricos , Animales , Bovinos , Enfermedades de los Bovinos/etiología , Enfermedades de los Bovinos/microbiología , Dinamarca/epidemiología , Masculino , Prevalencia , Factores de Riesgo , Salmonella/aislamiento & purificación , Salmonelosis Animal/etiología , Salmonelosis Animal/microbiología , Salmonella typhimurium/aislamiento & purificación
5.
Animal ; 5(6): 851-60, 2011 May.
Artículo en Inglés | MEDLINE | ID: mdl-22440024

RESUMEN

The restrictions on the use of antibiotic and anthelmintic treatments in organic pig farming necessitate alternative non-medical control strategies. Therefore, the antibiotic and parasite-reducing effect of a fructan-rich (prebiotic) diet of dried chicory was investigated in free-ranging piglets. Approximately half of 67 piglets from nine litters were experimentally infected with Ascaris suum and Trichuris suis in the suckling period (1 to 7 weeks of age) and 58 of the piglets were challenged daily with Eschericia coli O138:F8 for 9 days after weaning to induce weaning diarrhoea. The litters were fed either chicory (30% dry matter) or a control diet. The effect of chicory on intestinal helminths, intestinal microbiota, especially Bifidobacteria and Campylobacter spp. and E. coli post-weaning diarrhoea was assessed. The weight gain of the piglets was not impaired significantly by chicory. The intestinal A. suum worm burden was reduced by 64% (P = 0.034) in the chicory-fed piglets, whereas these same piglets had 63% more T. suis worms (P = 0.016). Feeding with chicory elicited no changes among the main bacterial groups in ileum according to terminal restriction fragment length polymorphism analysis. However, the terminal-restriction fragment (T-RF) 208 bp, which may belong to Lachnospiraceae, was stimulated by the chicory feed (P = 0.03), and T-RF 370 bp that matches Enterobacter belonging to the Enterobacteria was reduced (P = 0.004). In addition, chicory increased the level of Bifidobacteria (P = 0.001) and the faecal Campylobacter excretion level was transitorily reduced in chicory-fed piglets at 7 weeks of age (P = 0.029). Unfortunately, it was not possible to assess the effect of chicory on post-weaning diarrhoea as it did not develop. In conclusion, feeding piglets chicory around the time of weaning caused complex changes of the microbiota and parasite communities within the intestinal tract, and feeding piglets chicory may therefore serve as an animal-friendly strategy to control pathogens.

6.
Euro Surveill ; 15(4): 19471, 2010 Jan 28.
Artículo en Inglés | MEDLINE | ID: mdl-20122382

RESUMEN

Phage typing has for decades been useful as a phenotypical, definitive method for epidemiological characterisation of Salmonella Typhimurium. The system recommended by the World Health Organization (WHO) Collaborative Centre for phage typing of Salmonella has, however, become rather complex, and the present study illustrates the challenges of sufficient standardisation of the interpretation of lysis results to make sure that the same strain is assigned to the same phage type in different laboratories. Even though molecular typing methods will replace phenotypic characterisation methods in the future, it is our opinion that phage typing will remain for some time a useful tool to strengthen global Salmonella surveillance. Therefore, improved standardisation and quality assurance is essential to obtain a robust and harmonized method that allows comparison of results between laboratories.


Asunto(s)
Tipificación de Bacteriófagos , Brotes de Enfermedades , Intoxicación Alimentaria por Salmonella/microbiología , Salmonella typhimurium/clasificación , Tipificación de Bacteriófagos/normas , Dinamarca/epidemiología , Europa (Continente) , Humanos , Intoxicación Alimentaria por Salmonella/epidemiología , Organización Mundial de la Salud
7.
Epidemiol Infect ; 136(8): 1124-30, 2008 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-17877841

RESUMEN

In the investigation we report the occurrence and spread of multiresistant Salmonella Typhimurium phage type (DT) 104 in 174 known Danish animal herds infected between 1996 and 2003. We applied PFGE, plasmid analysis and spatio-temporal analysis for a description of the development and spread of infection among herds. The results showed that in Denmark DT104 are mainly spread by trade of animals; however, horizontal spread of DT104 is also important and the risk of an animal herd becoming infected increased with the proximity in time and space to a DT104-infected herd. Based on these results, it is recommended that specific actions are taken to limit the risk of horizontal spread of DT104 from newly infected herds.


Asunto(s)
Salmonelosis Animal/microbiología , Salmonelosis Animal/transmisión , Salmonella typhimurium/clasificación , Animales , Animales Domésticos/microbiología , ADN Bacteriano/análisis , Dinamarca/epidemiología , Transmisión de Enfermedad Infecciosa , Plásmidos , Salmonelosis Animal/epidemiología
8.
J Appl Microbiol ; 105(5): 1558-68, 2008 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-19146492

RESUMEN

AIMS: To investigate the transmission of Salmonella spp. between production animals (pigs and cattle) and wildlife on production animal farms in Denmark. METHODS AND RESULTS: In the winter and summer of 2001 and 2002, 3622 samples were collected from Salmonella-infected and noninfected herds of pigs and cattle and surrounding wildlife. Salmonella was detected in wildlife on farms carrying Salmonella-positive production animals and only during the periods when Salmonella was detected in the production animals. The presence of Salmonella Typhimurium in wild birds significantly correlated to their migration pattern and food preference. CONCLUSIONS: Salmonella was transmitted from infected herds of production animals (cattle and pigs) to wildlife that lived amongst or in close proximity to them. SIGNIFICANCE AND IMPACT OF THE STUDY: Salmonella in animal food products is associated with the occurrence of Salmonella in primary animal production. Strategies to control the introduction and spread of infection should include wildlife management, as the nearby wildlife may act as reservoirs for Salmonella spp. and/or may be passive carriers of the bacteria.


Asunto(s)
Animales Salvajes , Enfermedades de los Bovinos/epidemiología , Salmonelosis Animal/transmisión , Enfermedades de los Porcinos/epidemiología , Animales , Aves , Bovinos , Enfermedades de los Bovinos/microbiología , Dinamarca/epidemiología , Heces/microbiología , Incidencia , Insectos , Estudios Longitudinales , Roedores , Salmonelosis Animal/epidemiología , Salmonelosis Animal/microbiología , Salmonella enterica/clasificación , Salmonella enterica/aislamiento & purificación , Porcinos , Enfermedades de los Porcinos/microbiología
9.
J Appl Microbiol ; 103(3): 650-6, 2007 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-17714398

RESUMEN

AIMS: To analyse the relative importance of different biological and technical factors on the analytical sensitivity of conventional culture methods for detection of Salmonella Dublin in cattle faeces. METHODS AND RESULTS: Faeces samples collected from six adult bovines from different salmonella-negative herds were split into subpools and spiked with three strains of S. Dublin at a concentration level of c. 10 CFU g(-1) faeces. Each of the 18 strain-pools was divided into two sets of triplicates of four volumes of faecal matter (1, 5, 10 and 25 g). The two sets were pre-enriched with and without novobiocin, followed by combinations of culture media (three types) and selective media (two types). The sensitivity of each combination and sources of variation in detection were determined by a generalized linear mixed model using a split-plot design. CONCLUSIONS: Biological factors, such as faecal origin and S. Dublin strain influenced the sensitivity more than technical factors. Overall, the modified semi-solid Rappaport Vassiliadis (MSRV)-culture medium had the most reliable detection capability, whereas detection with selenite cystine broth and Mueller Kauffman tetrathionate broth combinations varied more in sensitivity and rarely reached the same level of detection as MSRV in this experiment. SIGNIFICANCE AND IMPACT OF THE STUDY: The study showed that for MSRV-culture medium and xylose lysine decarboxylase agar as the indicative medium, the sensitivity of the faecal culture method may be improved by focusing on the strain variations and the ecology of the faecal sample. Detailed investigation of the faecal flora (pathogens and normal flora) and the interaction with chemical factors may result in developing an improved method for detection of S. Dublin.


Asunto(s)
Enfermedades de los Bovinos/microbiología , Heces/microbiología , Inhibidores de Crecimiento , Salmonelosis Animal/microbiología , Salmonella enterica/aislamiento & purificación , Animales , Antibacterianos , Técnicas Bacteriológicas/métodos , Bovinos , Recuento de Colonia Microbiana , Medios de Cultivo , Heces/química , Metalotioneína 3 , Proteínas del Tejido Nervioso , Novobiocina , Salmonella enterica/efectos de los fármacos
10.
Microb Drug Resist ; 13(4): 289-94, 2007.
Artículo en Inglés | MEDLINE | ID: mdl-18184055

RESUMEN

The aims of the present study were to investigate at the farm-owner level the effect of prescribed tetracycline consumption in pigs and different Salmonella Typhimurium phage types on the probability that the S. Typhimurium was resistant to tetracycline. In this study, 1,307 isolates were included, originating from 877 farm owners, and data were analyzed using logistic regression. The analysis showed that both the S. Typhimurium phage type (p < 0.0001) and an increase in tetracycline consumption (p = 0.0007) were significantly associated with tetracycline resistance. In particular, the phage type was strongly associated with tetracycline resistance. A further analysis of data from the Danish Integrated Antimicrobial Resistance Monitoring and Research Programme (DANMAP) indicates that the tetracycline-susceptible phage types only slowly become tetracycline resistant, although tetracycline consumption more than doubled at the national level from 12,000-13,000 kg of active compound in 1996-1998 to 29,000 kg of active compound in 2004. Instead, tetracycline-resistant S. Typhimurium phage types became more prevalent. This suggests that the spread of already established or new resistant clones, rather than conversion of "old" well-established susceptible clones to resistant clones by uptake of resistance genes, explains most of the increased levels of tetracycline resistance in S. Typhimurium in Danish swine production in response to increased tetracycline consumption.


Asunto(s)
Tipificación de Bacteriófagos , Salmonella typhimurium/efectos de los fármacos , Porcinos/microbiología , Tetraciclina/administración & dosificación , Animales , Probabilidad , Salmonella typhimurium/clasificación , Resistencia a la Tetraciclina , Factores de Tiempo
11.
J Appl Microbiol ; 101(4): 883-90, 2006 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-16968300

RESUMEN

AIMS: Salmonella Typhimurium DT 104 is generally assumed to be spread by contact between live animals, e.g. by trading. The aim of the present study was to assess the importance of other routes of transmission in the dissemination of this bacterium. METHODS AND RESULTS: An outbreak among 14 cattle and pig herds located in a geographically narrow area in Denmark was investigated. Epidemiological information and disease history of the herds was obtained through interviews. Based on this, the hypothesis for horizontal spread was proposed, and these were confirmed by comparison of the pulsed field gel electrophoresis (PFGE) and the plasmid profiles of isolates obtained by continuous sampling over a period of almost 3 years. CONCLUSIONS: The study indicated that other routes might play an important role, than the trading of live animals, in the spread of S. Typhimurium DT 104 among livestock. SIGNIFICANCE AND IMPACT OF THE STUDY: Salmonella Typhimurium DT 104 infected herd might pose a significant risk to herds located within the same geographic area. In advising on how to avoid the spread of this bacterium, factors like person contacts, sharing of equipment and contaminated slurry should be focussed on in addition to infected animals.


Asunto(s)
Enfermedades de los Bovinos/transmisión , Microbiología Ambiental , Salmonelosis Animal/transmisión , Salmonella typhimurium , Enfermedades de los Porcinos/transmisión , Crianza de Animales Domésticos , Animales , Bovinos , Dermatoglifia del ADN , Dinamarca , Brotes de Enfermedades , Transmisión de Enfermedad Infecciosa , Electroforesis en Gel de Campo Pulsado , Genes Bacterianos , Estiércol/microbiología , Salmonella typhimurium/genética , Porcinos , Topografía Médica
12.
Vet Microbiol ; 116(1-3): 96-105, 2006 Aug 25.
Artículo en Inglés | MEDLINE | ID: mdl-16697127

RESUMEN

The occurrence and species distribution of thermophilic Campylobacter was investigated in organic outdoor pigs. An increased exposure of outdoor pigs to C. jejuni from the environment may cause a shift from a normal dominance of C. coli to more C. jejuni, which may imply a concern of reduced food safety. Bacteriological methods for determination of Campylobacter excretion level were combined with colony-blot hybridization and real-time PCR for specific detection of C. jejuni in pigs. Campylobacter was isolated from pigs (n=47), paddock environment (n=126) and wildlife (n=44), identified to species by real-time PCR and sub-typed by serotyping (Penner) and pulse-field gel electrophoresis (PFGE) genotyping. All pigs excreted Campylobacter (10(3)-10(7) CFU g(-1) faeces) from the age of 8-13-weeks old. C. jejuni was found in 29% of pigs in three consecutive trials and always in minority to C. coli (0.3-46%). C. jejuni and C. coli were isolated from 10% and 29% of the environmental samples, respectively, while crow-birds and rats harboured C. jejuni. Individual pigs hosted several strains (up to nine serotypes). The paddock environment was contaminated with C. coli serotypes similar to pig isolates, while most of the C. jejuni serotypes differed. C. jejuni isolates of different origin comprised few similar serotypes, just one identical genotype was common between pigs, environment and birds. In conclusion, the occurrence of C. jejuni varied considerably between the three groups of outdoor pigs. Furthermore, transfer of C. jejuni to the outdoor pigs from the nearby environment was not predominant according to the subtype dissimilarities of the obtained isolates.


Asunto(s)
Infecciones por Campylobacter/veterinaria , Campylobacter coli/aislamiento & purificación , Campylobacter jejuni/aislamiento & purificación , Enfermedades de los Porcinos/microbiología , Crianza de Animales Domésticos , Animales , Campylobacter coli/clasificación , Campylobacter jejuni/clasificación , Heces/microbiología , Microbiología del Suelo , Porcinos , Microbiología del Agua
13.
Vet Parasitol ; 139(1-3): 158-67, 2006 Jun 30.
Artículo en Inglés | MEDLINE | ID: mdl-16621287

RESUMEN

Concomitant infections with helminths and bacteria may affect the course and the resulting disease outcome of the individual infections. Salmonella, Oesophagostomum, Trichuris and Ascaris coexist naturally in pig herds in Denmark, and possible interactions were studied. Pigs in one experiment were trickle infected with low or moderate dose levels of Oesophagostomum spp. and challenge infected with S. Typhimurium. In another experiment, pigs were inoculated with S. Typhimurium followed by a challenge exposure to either Oesophagostomum, Trichuris or Ascaris. Enhancement of the Salmonella infection was not demonstrated in either experiment. The helminth effect on the pigs was modest and may explain the lack of influence on the Salmonella infection. A previous experiment with a larger Oesophagostomum infection level resulted in enhancement of the S. Typhimurium infection. A dose dependency of the interaction is therefore suggested. However, the relatively high worm burdens in the present study suggest that infection with these common pig helminths does generally not influence the course of concurrent S. Typhimurium infections under natural conditions.


Asunto(s)
Helmintiasis Animal/complicaciones , Parasitosis Intestinales/veterinaria , Salmonelosis Animal/complicaciones , Salmonella typhimurium/crecimiento & desarrollo , Enfermedades de los Porcinos/microbiología , Enfermedades de los Porcinos/parasitología , Animales , Ascaris/crecimiento & desarrollo , Ascaris/patogenicidad , Recuento de Colonia Microbiana/veterinaria , Heces/microbiología , Heces/parasitología , Helmintiasis Animal/epidemiología , Parasitosis Intestinales/complicaciones , Parasitosis Intestinales/epidemiología , Oesophagostomum/crecimiento & desarrollo , Oesophagostomum/patogenicidad , Recuento de Huevos de Parásitos/veterinaria , Dinámica Poblacional , Distribución Aleatoria , Salmonelosis Animal/epidemiología , Salmonella typhimurium/patogenicidad , Organismos Libres de Patógenos Específicos , Porcinos , Trichuris/crecimiento & desarrollo , Trichuris/patogenicidad
14.
Epidemiol Infect ; 134(1): 179-85, 2006 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-16409666

RESUMEN

Fifty-seven Salmonella Typhimurium strains isolated from poultry, swine and animal feed in Poland during the years 1979-1998 and 2000-2002 were analysed with conventional and molecular techniques. Antimicrobial resistance as well as multiresistance was found, respectively, in 80.1% and 56.1% of the isolates and most frequently among isolates from 2000-2002. Of several phage types noted, DT104 was prevalent among poultry, swine and feed isolates. DT104, U302 and non-typable strains had a multiple resistant profile (ACSSuT) due to the presence of class I integrons. Pulse-field gel electrophoresis of XbaI and BlnI digest showed high genomic similarity between the strains and confirmed clonal spread of S. Typhimurium infections. Plasmid profiling allowed further differentiation of the strains. We have, therefore, confirmed the appearance of S. Typhimurium DT104 showing genome integrated integron-mediated antimicrobial resistance in Poland. These findings are significant for public and animal health risks and document the dissemination of DT104 epidemic strains into new geographical regions.


Asunto(s)
Resistencia a Múltiples Medicamentos , Salmonelosis Animal/tratamiento farmacológico , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/patogenicidad , Alimentación Animal/microbiología , Animales , Farmacorresistencia Microbiana , Estudios Epidemiológicos , Humanos , Polonia/epidemiología , Aves de Corral , Enfermedades de las Aves de Corral/tratamiento farmacológico , Enfermedades de las Aves de Corral/epidemiología , Prevalencia , Salud Pública , Medición de Riesgo , Salmonelosis Animal/epidemiología , Salmonella typhimurium/clasificación , Salmonella typhimurium/genética , Serotipificación , Porcinos , Enfermedades de los Porcinos/tratamiento farmacológico , Enfermedades de los Porcinos/epidemiología
15.
J Appl Microbiol ; 99(2): 292-300, 2005.
Artículo en Inglés | MEDLINE | ID: mdl-16033460

RESUMEN

AIMS: To develop a real-time (rt) PCR for species differentiation of thermophilic Campylobacter and to develop a method for assessing co-colonization of pigs by Campylobacter spp. METHODS AND RESULTS: The specificity of a developed 5' nuclease rt-PCR for species-specific identification of Campylobacter jejuni, Campylobacter coli, Campylobacter lari, Campylobacter upsaliensis and of a hipO gene nucleotide probe for detection of C. jejuni by colony-blot hybridization were determined by testing a total of 75 reference strains of Campylobacter spp. and related organisms. The rt-PCR method allowed species-specific detection of Campylobacter spp. in naturally infected pig faecal samples after an enrichment step, whereas the hybridization approach enhanced the specific isolation of C. jejuni (present in minority to C. coli) from pigs. CONCLUSIONS: The rt-PCR was specific for Campylobacter jejuni, C. coli, C. lari, and C. upsaliensis and the colony-blot hybridization approach provided an effective tool for isolation of C. jejuni from pig faecal samples typically dominated by C. coli. SIGNIFICANCE AND IMPACT OF THE STUDY: Species differentiation between thermophilic Campylobacter is difficult by phenotypic methods and the developed rt-PCR provides an easy and fast method for such differentiation. Detection of C. jejuni by colony hybridization may increase the isolation rate of this species from pig faeces.


Asunto(s)
Campylobacter/aislamiento & purificación , Heces/microbiología , Hibridación de Ácido Nucleico/métodos , Reacción en Cadena de la Polimerasa/métodos , Animales , Campylobacter/genética , Campylobacter coli/genética , Campylobacter coli/aislamiento & purificación , Campylobacter jejuni/genética , Campylobacter jejuni/aislamiento & purificación , Campylobacter lari/genética , Campylobacter lari/aislamiento & purificación , Campylobacter upsaliensis/genética , Campylobacter upsaliensis/aislamiento & purificación , Sondas de ADN/genética , ADN Bacteriano/genética , Genes Bacterianos/genética , Especificidad de la Especie , Porcinos
16.
Prev Vet Med ; 69(3-4): 213-28, 2005 Jul 12.
Artículo en Inglés | MEDLINE | ID: mdl-15907571

RESUMEN

A pilot study was carried out on a Danish swine farm infected with multi-resistant Salmonella Typhimurium DT104 (MRDT104). We aimed to (1) investigate to which degree the decline of Escherichia coli and Salmonella in swine slurry applied to farmland depended on the application method; (2) estimate the survival times of E. coli and Salmonella in the soil surface following deposition of naturally contaminated pig slurry; and (3) simulate survival of Salmonella in different infection levels using E. coli data as input estimates. Slurry was deposited by four different methods: (1) hose applicator on black soil followed by ploughing and harrowing; (2) hose applicator on black soil followed only by harrowing; (3) hose applicator on a field with winter-wheat seedlings without further soil treatment; (4) slurry injector on a field with winter-wheat seedlings without further soil treatment. E. coli and Salmonella could not be detected at all in soil following treatment 1. Following the other treatments, E. coli was not detected in soil samples after day 21 and Salmonella was no longer detected after day 7. Simulation results showed that clinical (4 log CFU g(-1)) and sub-clinical Salmonella levels (2500 CFU g(-1)) would fall below the detection limit within 10 or 5 days, respectively. Analysis of samples from 62 Danish MRDT104-infected swineherds showed that nearly 75% of these herds had low levels of MRDT104 (< 10 CFU g(-1)) in their slurry. Our results show that ploughing and harrowing of soil amended with contaminated pig slurry was an effective means to reduce environmental exposure to E. coli and Salmonella on this clay-soil farm.


Asunto(s)
Escherichia coli/crecimiento & desarrollo , Estiércol/microbiología , Salmonella typhimurium/crecimiento & desarrollo , Microbiología del Suelo , Porcinos/microbiología , Silicatos de Aluminio , Animales , Arcilla , Simulación por Computador , Dinamarca , Infecciones por Escherichia coli/microbiología , Infecciones por Escherichia coli/veterinaria , Modelos Biológicos , Proyectos Piloto , Salmonelosis Animal/microbiología
17.
J Food Prot ; 68(4): 722-7, 2005 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-15830662

RESUMEN

The prevalence and diversity of Campylobacter jejuni was investigated in pig herds on farms with and without cattle or poultry production. A bacteriological screening of pig cecal samples from 247 finisher herds was carried out at the slaughter-house. Subsequently, a follow-up study was conducted in 24 herds (either with or without prior C. jejuni isolation from pigs) in which fecal samples were collected from pigs and, if present, cattle and poultry. Samples were analyzed for presence of Campylobacter, and subsequent analysis included species identification, serotyping, and, for selected strains, pulsed-field gel electrophoresis typing. In the slaughterhouse screening, C. jejuni was isolated from pigs in 21 (8.5%) herds, but no significant difference in prevalence was found between herd types (pigs, pigs and cattle, pigs and poultry). At the slaughterhouse, C. jejuni and Campylobacter coli prevalence in pigs was 2.3 and 90.1%, respectively. In the follow-up study, herd prevalence of C. jejuni was 8.3%, whereas C. jejuni and C. coli were isolated from 0.8 and 92.0% of pigs, respectively. In mixed production herds, C. jejuni predominated in cattle (42.7%) and poultry (31.6%), whereas C. jejuni was only isolated from 1.3 to 2.5% of pigs in these herds. There were no significant differences in C. jejuni or C. coli prevalence in pigs, cattle, and poultry between herds with and without prior C. jejuni isolation at the slaughterhouse. Pulsed-field gel electrophoresis typing did not yield evidence of C. jejuni transmission between cattle or poultry and pigs in mixed production herds. In contrast, pulsed-field gel electrophoresis analysis showed indistinguishable serotypes of C. coli in pigs and cattle in two herds. Verification of C. jejuni-positive pig samples showed that individual pigs can excrete high levels of C. jejuni and that mixed infection with C. jejuni and C. coli was common in C. jejuni-positive pigs. The results of our study suggest that transmission of C. jejuni between pigs and cattle or poultry in mixed production herds occurs infrequently. Detection of indistinguishable C. coli isolates in two herds, however, might indicate the existence of low-level transmission between pigs and cattle in herds of mixed production.


Asunto(s)
Infecciones por Campylobacter/veterinaria , Campylobacter jejuni/aislamiento & purificación , Enfermedades de los Bovinos/epidemiología , Enfermedades de las Aves de Corral/epidemiología , Enfermedades de los Porcinos/epidemiología , Mataderos , Animales , Campylobacter/crecimiento & desarrollo , Campylobacter/aislamiento & purificación , Infecciones por Campylobacter/epidemiología , Infecciones por Campylobacter/transmisión , Campylobacter jejuni/clasificación , Campylobacter jejuni/crecimiento & desarrollo , Bovinos , Enfermedades de los Bovinos/transmisión , Ciego/microbiología , Transmisión de Enfermedad Infecciosa/veterinaria , Contaminación de Alimentos/prevención & control , Aves de Corral , Enfermedades de las Aves de Corral/transmisión , Prevalencia , Serotipificación/veterinaria , Especificidad de la Especie , Porcinos , Enfermedades de los Porcinos/transmisión
18.
J Microbiol Methods ; 55(1): 249-55, 2003 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-14500016

RESUMEN

The aim was to investigate the effect of addition of Novobiocin to the non-selective buffered peptone water (BPW) for pre-enrichment of Salmonella in connection with plating on modified semisolid Rappaport-Vassiliadis (MSRV). In a semi-quantitative study, the level of Salmonella following pre-enrichment of 32 presumably naturally contaminated swine fecal samples were assessed for BPW with and without addition of Novobiocin (22 microg/ml). In another experiment, a total of 400 swine fecal samples were screened for the presence of Salmonella spp., in order to compare the performance of the non-selective pre-enrichment broth with BPW made semi-selective by addition of Novobiocin. The semi-quantitative assessment of the Salmonella level showed that addition of Novobiocin in the pre-enrichment step on average increased the level of Salmonella 1.2 log dilution steps. When growth was scored at five levels, 90 samples opposed to 50 yielded a strong positive reading (+++) when Novobiocin was applied. Growth was on average 0.3 scores higher when pre-enriched with Novobiocin. The difference in growth score medians of the two methods was highly significant (Sign test; p<0.001). Despite the increased sensitivity, 13 culture-positive samples were missed when using the Novobiocin-containing BPW. In conclusion, a simple addition of Novobiocin in the BPW pre-enrichment step of fecal samples may facilitate reading and thereby detection of Salmonella on MSRV. The increase of Salmonella in the semi-quantitative study may be caused by a reduction in the number of competitive microorganisms.


Asunto(s)
Heces/microbiología , Novobiocina/farmacología , Salmonella/aislamiento & purificación , Porcinos/microbiología , Animales , Medios de Cultivo , Salmonella/efectos de los fármacos
19.
Ugeskr Laeger ; 163(41): 5677-8, 2001 Oct 08.
Artículo en Danés | MEDLINE | ID: mdl-11665472

RESUMEN

Whereas the overall incidence of human Salmonella infections in Denmark has fallen during the past three years, the number of infections with multidrug-resistant Salmonella Typhimurium definitive type 104 (DT104) has risen. We report five cases of human infection with DT104 in patients living on farms, in which cattle and mixed herds of cattle and pigs were infected with DT104. Transmission from the animals to the patients in the cases described is likely to have occurred. These cases emphasize the risky of infection through contact with animals infected with DT104.


Asunto(s)
Gastroenteritis/microbiología , Intoxicación Alimentaria por Salmonella/microbiología , Salmonella enterica/clasificación , Salmonella typhimurium/clasificación , Adolescente , Adulto , Animales , Bovinos , Niño , Dinamarca , Femenino , Gastroenteritis/tratamiento farmacológico , Humanos , Masculino , Intoxicación Alimentaria por Salmonella/tratamiento farmacológico , Intoxicación Alimentaria por Salmonella/transmisión , Serotipificación , Porcinos
20.
Berl Munch Tierarztl Wochenschr ; 114(9-10): 323-6, 2001.
Artículo en Inglés | MEDLINE | ID: mdl-11570168

RESUMEN

The Danish Salmonella Surveillance and Control Programme for pigs operates at all stages of the production chain and has been applied nationally since 1995. Due to the program the level of Salmonella in Danish pork has declined from 3.5% in 1993 to 0.7% in the year 2000. Simultaneously, the number of human cases with salmonellosis due to pork has declined from approximately 1,144 in 1993 to 166 in 2000. In year 2001, the programme has been improved at a number of stages. A new classification scheme for the serological surveillance of finisher herds has been developed. The individual test cut-off in the mix-ELISA has been reduced to 20 OD%. Only herds producing more than 200 finishers/year are sampled. Based on the serological result from the last 3 months a new weighted salmonella index is calculated: The Danish Bacon and Meat Council has agreed on a new stricter penalty system. Level 2 and 3 herds get a penalty of 2% and 4% of the value per slaughter carcass, respectively. A new method of Salmonella testing on carcasses has been introduced; 5 carcasses per slaughter day are swabbed at 3 defined areas at 100 cm2 for each sample. This method is more sensitive than the one used previously. Herds infected with multiresistant Salmonella Typhimurium DT104 have to follow special restrictions. These include a requirement for a herd intervention plan, restriction on livestock trade, and a requirement for special slurry handling. Carcasses from DT 104 herds must be heat-treated or decontaminated with hot water.


Asunto(s)
Intoxicación Alimentaria por Salmonella/prevención & control , Salmonelosis Animal/prevención & control , Salmonella/aislamiento & purificación , Enfermedades de los Porcinos/prevención & control , Mataderos , Animales , Dinamarca/epidemiología , Desinfección/métodos , Contaminación de Alimentos , Manipulación de Alimentos , Humanos , Incidencia , Factores de Riesgo , Intoxicación Alimentaria por Salmonella/epidemiología , Salmonelosis Animal/epidemiología , Porcinos , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/transmisión , Zoonosis
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