Salmonella enterica infections in Spanish swine fattening units

The present study is the first conducted in Spain to estimate the bacteriological herd prevalence of Salmonella enterica in fattening units and to describe the Salmonella serovar diversity on these farms using a sample representative of the entire swine population. For this purpose, 10 faecal samples were collected from 10 different pens containing pigs close to market weight in a total of 232 fattening units. Total sample size was proportionally distributed according to the fattener census in each of the regions of the country and all the samples were examined by culture of 25 g of faecal material. One hundred (43.1%) farms had at least one Salmonella-positive sample (95% CI: 37-49.1%). Salmonella enterica was detected in 290 (12.5%) pooled faecal floor samples (95% CI: 11.2-13.8%). The apparent herd prevalence of salmonellosis was similar among multi-site, finishing and farrow to finish farms. Overall, 24 different serovars were identified, with S. Typhimurium, S. Rissen and S. Derby being the most common both at herd and sample level. Results of phage typing were available for the 91 isolates of S. Typhimurium. A total number of 10 different phage types were identified, with DT 193 being the most frequent. Phage types DT 104, DT 104b and DT U302, which have been associated with several multi-resistant patterns, accounted for 23% and 29% of the Typhimurium total isolates or Typhimurium infected farms respectively.     

Impact of commercial pre-harvest processes on the prevalence of Salmonella enterica in cull sows

This study examined the impact of transport and lairage on Salmonella enterica prevalence in cull sows held under commercial conditions. Five sampling periods over 10 weeks yielded 181 selected culls. Fecal sample positives at the farm and collection point were 2% and 3%, respectively. After transport and lairage 41% of cull sows yielded S. enterica in one or more sampled tissues. A significant increase in cecal isolations was observed. The two S. enterica serotypes found at the production site and collection point were found at the abattoir, as were 13 other serotypes. Transport and lairage practices may contribute to an increased S. enterica contamination immediately prior to slaughter.     

Evaluation of stocking density and subtherapeutic chlortetracycline on Salmonella enterica subsp. enterica shedding in growing swine

The objective of this research was to determine the effect of stocking density and inclusion of subtherapeutic chlortetracycline in the diet on Salmonella fecal prevalence and antimicrobial resistance in growing swine. A 2 x 2 factorial design was employed on a privately owned commercial swine farm. Four finisher rooms were included in the study. Two of the rooms received 50 g/tonnes of chlortetracycline in the ration, two rooms received no antimicrobials in the feed. In each room, alternate pens were assigned to either high stocking density (0.60 m2/pig) or low stocking density (0.74 m2/pig). Pigs were placed in the finisher rooms at 10 weeks of age and followed for 6 weeks. Individual fecal samples were collected from the floors of each pen and cultured once weekly. Antimicrobial resistance phenotypes were determined. Data were analyzed using multilevel, multivariable logistic regression. Pigs fed chlortetracycline were at increased odds (OR 6.88, 95% CI 2.77-17.12) to shed Salmonellae. No other associations between treatments (CTC and stocking density) and Salmonella prevalence or reduced susceptibility to antimicrobials were identified. Variance in the odds of a fecal sample to be positive was distributed mostly at the lowest level, the individual fecal sample. The increased risk of shedding associated with inclusion of subtherapeutic chlortetracycline in swine diets is discordant with previous results by our group, suggesting farm or strain specific factors may impact this association. Understanding this risk may provide a potential intervention for controlling Salmonella pre-harvest.     

Comparison of fecal culture and Danish Mix-ELISA for determination of Salmonella enterica subsp. enterica prevalence in growing swine

In the USA, control of food-borne salmonellosis associated with meat consumption has been predominantly focused at slaughter and processing. It is expected that standards at slaughter and processing will become more stringent, creating pressure to reduce prevalence of Salmonella-positive food animals through on-farm interventions. The aim of this study was to compare traditional fecal culture and the Danish Mix-ELISA (DME) for determination of Salmonella prevalence pre-harvest in swine. In Trial 1, five cohorts of individually identified pigs were longitudinally sampled during the growing period to compare the kinetics of prevalence as estimated by fecal culture and the DME. In Trial 2, the correlation between fecal prevalence and seroprevalence was estimated pre-marketing in 49 groups of pigs. In Trial 1, fecal prevalence and seroprevalence showed similar kinetics, with a tendency of a higher OD% cut-off to more closely approximate fecal prevalence. In Trial 2, correlations between fecal culture and the DME were 0.40, 0.36, 0.43, and 0.43 (p<0.001) for OD% cut-offs > or =10, 20, 30, and 40, respectively. Based on these results, a higher OD% cut-off would be recommended if more approximate estimation of fecal prevalence is desired and longitudinal sampling would be suggested for evaluating the impact of on-farm interventions for Salmonella reduction whether utilizing fecal culture or the DME. Further evaluation of the impact of Salmonella serovar present on farms on seroprevalence and the relationship of on-farm seroprevalence with food safety risk are needed prior to utilizing the DME for pre-harvest Salmonella diagnostics in the US swine herd.     

The effect of fecal sample weight on detection of Salmonella enterica in swine feces.

The effect of different fecal sample weights on the detection of Salmonella enterica in swine feces was examined. Sample weights evaluated were rectal swabs and fecal samples weighing 1 g, 10 g, and 25 g. Comparisons were made on matched fecal samples obtained from individual pigs housed on 2 commercial swine farms in North Carolina. Relative sensitivity (number of positive pigs per fecal weight category/number positive in all weight categories) increased (P < 0.001) with fecal sample weight, and ranged from 9% for rectal swabs to 78% for 25-g samples. Stomaching of fecal samples did not affect detection of S. enterica. These observations demonstrate that fecal sample weight can markedly influence estimates of prevalence of S. enterica in epidemiologic studies. Failure to consider the imperfect sensitivity of bacterial culture in the design and interpretation of epidemiologic studies will lead to underestimation of prevalence and reduced power to detect the presence of S. enterica-infected herds.     

Longitudinal study of Salmonella enterica in growing pigs reared in multiple-site swine production systems

Intensive longitudinal investigations of breeding and growing pig populations in two multiple-site swine production systems were conducted in NC, USA. Five cohorts of sows and individually identified growing pigs from their litters were serially sampled in order to determine the prevalence and serotypes of Salmonella enterica in each stage of production based on fecal culture. In addition to fecal samples, feed and environmental samples were obtained. Fifteen different serotypes were isolated from the two systems, the most frequently isolated serotypes were S. typhimurium var Mbandaka and S. typhimurium var Copenhagen. Pig prevalence estimates ranged from 0 to 48.1%. Environmental contamination was frequently encountered despite cleaning and disinfection. Feed was rarely (2/800, 0.25%) identified as S. enterica positive. We observed highly variable patterns of S. enterica prevalence and serotype profiles within cohorts over time and among cohorts within systems. These observations indicate that point estimates of S. enterica prevalence and serotypes cannot be considered as reliable indicators of the S. enterica status of farms, and that uncontrolled studies of interventions to control S. enterica may yield misleading results. These findings are critical to the design of epidemiological studies of S. enterica on swine farms and may suggest that cohort level, as opposed to farm or company level events or management practices, may be important as potential risk factors for S. enterica fecal shedding in market age pigs.     

Assessment of antibiotic resistance phenotype and integrons in Salmonella enterica serovar Typhimurium isolated from swine

Salmonella enterica serovar Typhimurium (S. Typhimurium) isolated and identified from swine were subjected for the analysis of antibiotic resistance pattern and clinically important class 1 and 2 integrons. In addition, S. Typhimurium isolates exhibiting ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, tetracycline and florfenicol (ACSSuTF) resistance pattern as described in most Salmonella enterica serotype Typhimurium definitive type 104 (DT104) were characterized by polymerase chain reaction. All the isolates were resistant to more than four antibiotics and showed the highest resistance to streptomycin (94.1%), followed by tetracycline (90.1%), ampicillin (64.7%), chloramphenicol (56.8%) and gentamicin (54.9%). MIC value for the ten isolates ranged between 0.125-2 mug/ml for ciprofloxacin. Among the beta-lactams used, only one of the isolate exhibited resistance to ceftiofur (MIC 8 microg/ml). Sixty eight percent of these multi drug resistance (MDR) S. Typhimurium isolates carried clinically important class 1 integron with 1kb (aadA) and/or 2kb (dhfrXII-orfF-aadA2) resistance gene cassettes. This study reports the increasing trend of multi drug resistance (MDR) S. Typhimurium with clinically important class 1 integron in pigs. In addition, emergence of the ACSSuTF-type resistance in S. Typhimurium PT other than DT104 may limit the use of resistance gene markers in its detection methods by PCR.     

Early epithelial invasion by Salmonella enterica serovar Typhimurium DT104 in the swine ileum

Salmonella enterica serovar Typhimurium is an important intestinal pathogen in swine. This study was performed to document the early cellular invasion of Salmonella serovar Typhimurium in swine ileum. Ileal gut-loops were surgically prepared in ten 4- to 5-week-old mixed-breed pigs and inoculated for 0-60 minutes. Loops were harvested and prepared for both scanning and transmission electron microscopy (SEM and TEM, respectively). Preferential bacterial adherence to microfold cells (M cells) was seen within 5 minutes, and by 10 minutes bacterial invasion of the apical membrane was seen in M cells, goblet cells, and enterocytes. This multicellular invasion was observed throughout the course of infection. In addition, SEM revealed a specific affinity of Salmonella serovar Typhimurium to sites of cell extrusion. Using TEM, bacteria in these areas were focused in the crevices formed by the extruding cell and the adjacent cells and in the cytoplasm immediately beneath the extruding cell. Our results suggest that early cellular invasion by Salmonella serovar Typhimurium is nonspecific and rapid in swine. Furthermore, the combination of SEM and TEM data suggests that Salmonella serovar Typhimurium may use sites of cell extrusion as an additional mechanism for early invasion.     

A longitudinal study of Salmonella enterica infections in high-and low-seroprevalence finishing swine herds in The Netherlands

The purpose of this investigation was to study the incidence and course of Salmonella infections in finishing pig herds in order to asses the stability of a given Salmonella herd status. Five low- and 7 high-seroprevalence herds were followed for seven sampling rounds. Each round, blood and faecal samples were tested in an indirect ELISA and by bacteriological culturing, respectively. In high-seroprevalence herds a positive Salmonella status was an indication of a long-term problem and the status was relatively stable over time. The herds experiencing clinical salmonellosis were not necessarily the herds with the highest seroprevalence. It is possible to deliver sero-negative finishers to the slaughterhouse, even though these pigs were seropositive as growers. In three out of five low-prevalence herds, major infection incidents occurred, indicating that changes in the Salmonella status should be anticipated. Low-prevalence herds can remain negative over a longer period of time as a result feeding a complete liquid feed containing fermented by-products.     

Testing an ecological model for transmission of Salmonella enterica in swine production ecosystems using genotyping data

An ecological model for transmission of Salmonella enterica in swine production ecosystems was developed, identifying host species, environmental reservoirs, and temporal, spatial, and functional (i.e., stage of production) dimensions. It was hypothesized that transmission was most likely within spatial and functional compartments, between hosts of the same species and abiotic compartments of the same type. Eighteen swine production systems in Illinois, USA, were sampled in four collection cycles (1998, 1999, 2000, 2003). There were 11,873 samples collected, including feces from swine and other mammals and birds, and samples from insects, pen floors, boots, feed, and water. The 460 Salmonella isolates obtained were genotyped using repetitive sequence PCR with three primers—REP, BOX, and ERIC. All isolates from 2000 and 2003 were serotyped, as well as a subsample from 1998 and 1998. Genetic relatedness was estimated from the similarity of fragmentation patterns after gel electrophoresis of PCR products. Cluster analysis identified genetically related isolates. Linking of isolates in tight clusters (similarity ≥85%) was viewed as evidence for transmission. Five farms had a sufficient number of tight clusters for hypothesis testing. The factors most differentiating isolates genetically were farm of origin and time of sampling. Isolates were also differentiated genetically by site, building, room, and pen. There was no consistent association of genotype with stage of production or host/environment reservoir. Serotype analysis confirmed that Salmonella lineages were differentiated by visit and site. Thus, Salmonella transmission was primarily over short distances, i.e., within the same pen or room, with some transmission between rooms and buildings on the same site, but with limited transmission between sites. Transmission was observed across a variety of ecological niches represented by different host species and environmental reservoirs. Genetic differences over time reflected multiple introductions into the ecosystem of different Salmonella genotypes, as well as evolutionary changes within lineages. Intervention strategies to reduce Salmonella prevalence within swine production ecosystems would be best targeted at maintaining spatial barriers to transmission, whereas intervention targeted at specific biological hosts or environmental reservoirs is less likely to be effective.     

Culture methods differ on the isolation of Salmonella enterica serotypes from naturally contaminated swine fecal samples.

Four culture methods (A, B, C, and D) were comparatively evaluated for their ability to isolate Salmonella enterica from pooled swine fecal samples (n = 100). None of the methods was able to isolate Salmonella from all positive samples. The relative sensitivity of the culture methods evaluated was 82%, 94%, 95%, and 78% for methods A, B, C, and D, respectively. The comparison of sensitivities showed that methods B and C performed significantly better (P < 0.05) than methods A and D. Although relative sensitivities of methods B and C were equal, from the 89 positive samples concomitantly detected by both, 35 (39.3%) had different serotypes (no match) isolated by each method. On the basis of the results of this study, it was concluded that culture methods differ on the isolation of S. enterica serotypes from naturally contaminated swine fecal samples. Depending on the objective(s) of investigations on the ecology and epidemiology of S. enterica in swine populations, a method or a combination of methods should be considered for more reliable results.     

Risk factors associated with Salmonella enterica prevalence in three-site swine production systems in North Carolina, USA

The goal of this study was to identify risk factors associated with increased fecal shedding of Salmonella enterica (SE) in groups of market swine reared in large three-site production units. We conducted an intensive, long-term investigation of potential management and environmental risk factors operating during the growing phase of pig production. Data regarding finisher site characteristics, biosecurity protocols, group growth performance, medication usage, and environmental temperature were collected. Results indicate that SE infection is common. Risk factors were identified at both the finisher site and group level. Biosecurity and hygiene practices (absence of a toilet, more than 2 people present at a finisher site daily, and other domestic species at the site), environmental temperature (winter and spring seasons, increased temperature variability, and below median high temperature the day of sampling), and production performance (above median feed conversion) were associated with elevated SE prevalence. In addition, an association between the floor space allowances per pig at the time of sampling (a measure of the number of pigs sold prior to sampling) was identified, with greater space allowance associated with decreased prevalence. The results of this study identify potential management practices for evaluation for SE control and suggest caution in interpretation of fecal culture results when sampling from different marketing groups in swine production systems.     

四环素耐药基因在鸡源沙门氏菌中的分布和传播

试验分析了84株鸡源沙门氏菌分离株的四环素耐药性,用PCR方法检测四环素耐药基因在分离株中的分布情况。结果显示,四环素耐药率为49%(41/84),鸡白痢和鸡伤寒沙门氏菌仅携带tet(A)基因(23/23),肠炎沙门氏菌和德尔卑沙门氏菌携带tet(A)(8/18)、tet(B)(17/18)或tet(G)(10/18)三种基因,tetC基因在这些沙门氏菌中都没有检测到。该类基因多数位于结合性质粒上,但是不在整合子范围内。     

对利用发酵床技术养猪的几点看法

随着养猪业的发展和猪群规模的不断扩大,猪粪尿的排放量也随之增加,尤其是规模化养猪场排泄的大量粪尿对生态环境造成的污染问题,迫切需要解决。发酵床养猪是一种环保、安全、有效的生态养猪法,是养猪业可持续发展的新模式,可以解决目前面临的污染问题。     

Distribution and function of plasmids in Salmonella enterica

Plasmids of Salmonella enterica vary in size from 2 to more than 200 kb. The best described group of plasmids are the virulence plasmids (50-100 kb in size) present in serovars Enteritidis, Typhimurium, Dublin, Cholerae-suis, Gallinarum, Pullorum and Abortus-ovis. They all encode spvRABCD genes involved in intra-macrophage survival of Salmonella. Another group of high molecular weight plasmids are plasmids responsible for antibiotic resistance. Since most of these plasmids are conjugative, besides storage of genetic information, they contribute to the spread of genes in bacterial populations. The low molecular weight plasmids are the last group of plasmids found in S. enterica. Some of them have been shown to increase resistance to phage infection due to the presence of restriction modification systems. Despite limited knowledge on their function, their presence or absence is frequently used for strain differentiation in epidemiological studies.     

Distribution of integrons and SGI1 among antibiotic-resistant Salmonella enterica isolates of animal origin

In Salmonella enterica, resistance to antibiotics can be caused by the presence of SGI1, transposons or conjugative plasmids. In this study we were interested in the relative contribution of these genetic elements to the antibiotic resistance of S. enterica isolates collected within a single year in the Czech Republic from animal sources. Altogether 123 antibiotic-resistant isolates belonging to 16 different S. enterica serovars were classified into 3 groups according to the presence of SGI1 and the presence of integrons. The first group consisted of 62 strains in which neither SGI1 nor class 1 integron was detected. A high diversity among serovars and resistance phenotypes was found in this group. The second group consisted of 56 strains positive for both the SGI1 and class 1 integron, out of which 55 belonged to serovar Typhimurium and one to a nonmotile serovar [4,12] which harboured the SGI1-B variant. The third group comprised five strains which were positive for class 1 integron but negative for the SGI1. Sequencing of the integrons in these isolates identified integron with sat1 and aadA1 gene cassettes in S. Sandiego and S. Pullorum, dfrA1 and aadA1 gene cassettes in S. Typhimurium integron, and aadA21 gene cassette in S. Braenderub and S. Zanzibar.     

Antimicrobial susceptibility and multi-drug resistance of Salmonella enterica subspecies enterica serovars in Sudan

This study was undertaken to determine the antimicrobial resistance patterns of Salmonella enterica subspecies enterica recovered from human, food, water, and animal samples collected in Khartoum State, Sudan. A total of 64 Salmonella isolates belonging to 28 different serovars were tested for their susceptibility to 13 antimicrobial agents. The majority of isolates (98.4 %) were resistant to at least one antimicrobial agent. Isolates were frequently resistant to ampicillin (90.6 %), cephalexin (50.0 %), nalidixic acid (25.0 %), streptomycin (21.9 %), kanamycin (18.8 %), gentamicin (17.2 %), and co-trimoxazole and trimethoprim (12.5 %). The most common pattern of multiple drug resistance included resistance to ampicillin and cephalexin. Most isolates were sensitive to chloramphenicol (98.4 %), ciprofloxacin (93.8 %), and norfloxacin (90.6 %). Two chicken- and the two human-origin S. Kentucky isolates were resistant to both ciprofloxacin and norfloxacin. All S. Kentucky isolates and the one S. Rissen isolate demonstrated multi-drug resistance. The results indicate the significance of multi-drug-resistant Salmonella serovars isolated from chickens and other animals and foods as sources for multi-drug-resistant Salmonella in humans in Sudan.     

Dunging gutters filled with fresh water in finishing barns had no effect on the prevalence of Salmonella enterica on Brazilian swine farms

Our aim was to assess the importance of dunging gutters filled with water in finishing barns for the prevalence of pigs shedding Salmonella enterica. Some finishing barns in Brazil are provided with a dunging-gutter system which consists of a continuous water flow at the back of solid-floored adjacent pens. Because there is transfer of faecal material between adjacent pens by water in this system and the faecal-oral route of transmission is so important for enteric pathogens, we tested the hypothesis that the presence of this kind of dunging-gutter system in finishing barns affects the prevalence of slaughter-age pigs shedding salmonella organisms in their faeces. The cross-sectional study was conducted on six farms each having barns with and barns without a dunging-gutter system. Breeding, management, nutritional and seasonal factors were similar in both barns on each farm. The two systems did not differ in prevalence of pigs shedding salmonella organisms. Five S. enterica serotypes were isolated: S. Agona, S. Javiana, S. Rissen, S. Sandiego and S. Senftenberg.