Studies on the interaction between Salmonella enterica ser. Typhimurium and intestinal helminths in pigs

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.     

Longitudinal serological responses to Salmonella enterica of growing pigs in a subclinically infected herd

A longitudinal survey was conducted in France in a subclinically Salmonella-infected farrow-to-finish pig farm to describe the time-course of the serological response to Salmonella enterica in growing pigs. We used three batches of sows and their corresponding litters (n = 31 litters). Among these, 256 pigs randomly selected and individually identified were followed from the first week of age until slaughter. Serial individual blood samples were submitted to indirect Salmonella-ELISA testing. Salmonella shedding was investigated by bacteriological testing of faecal material regularly collected from the sows and pigs and by environment swabs taken from the pens. Caecal contamination was checked at the slaughterhouse. Information about litter composition (filiation), location of the pigs in post-weaning and fattening pens, sanitary events, sex and body weights was recorded. 11.6% of the pigs shed S. enterica; 52% of pigs seroconverted before slaughter. The age-related variation of the natural logarithm of calibrated optical densities (COD) of pigs was described with two linear mixed models. From 8 to 61 days of age, the decrease in COD with age was fitted with a model including random effects of the animal and the dam on the intercept and slope, a batch random effect on the intercept and an individual birth-weight fixed effect on the intercept. The dam random effect was explained by the parity of the sow, Salmonella shedding by the sow during the farrowing phase and the value of the optical density of colostrum collected at parturition. A second model fitting the increase in COD from 61 days of age until slaughter included the random effect on intercept of the batch and the random effects on slope and intercept of the animal, the dam and the pen in which the followed animals were located during the fattening phase and the environmental contamination as fixed effect. In this second model, no relation was found between individual slaughter-bacteriological results and increasing COD values. Considering seroconversion time between 61 days of age and slaughter, survival function were constructed using the Cox proportional-hazards model. Both methods suggested that seroconversions generally occurred during the last third of the fattening phase (from 140 days of age to slaughter), while shedding was observed during the first half of the fattening period. The fitted models suggest the existence of clusters (such as pen and litter of origin).     

Resistance of Oesophagostomum spp. in pigs to pyrantel citrate

This study was undertaken to determine whether anthelmintic-resistant Oesophagostomum spp. populations occur in Danish swine herds. A controlled field trial on selected sow herds suggested resistance to pyrantel citrate in a herd where faecal egg count depression in response to treatment was only 28.3%. This was confirmed by subsequent experimental infection of pigs where the suspected resistant Oesophagostomum isolate was compared with a susceptible worm isolate. After treatment with the recommended dose of the drug, worm burdens of the suspected isolate were only reduced by 42.6% (p greater than 0.05) in contrast to the susceptible isolate, which was reduced by 94.6% (p less than 0.01). Differential counts of the resistant isolate suggested that O. dentatum might be more resistant than O. quadrispinulatum. The resistant isolate originated from a herd, where the anthelmintic had been administered four times a year over a period of 7 years. The potential for development of anthelmintic resistance in Oesophagostomum spp. under management practices in our country is discussed in the light of the current views on predisposing factors.     

Dose related mucosal hyperplasia induced by Oesophagostomum dentatum infection in pigs.

The present work was undertaken to examine the effects of 3 different population densities of Oesophagostomum dentatum upon the development of worm induced mucosal changes in the colon following single infections. Groups of pigs were infected with single doses of 2000 (low dose), 20,000 (medium dose) or 200,000 (high dose) infective larvae, respectively. A total of 18 infected pigs (6 from each group) were examined for histopathological changes together with 3 helminth-free control pigs. There was a dose related difference in the intensity of colonic lesions; and using morphometry it was observed that the mucosal crypts of pigs in the high dose group were significantly longer than those in the 2 other groups. These differences disappeared by day 25 after infection despite the presence of larvae in the mucosa of the high dose group. This phenomenon may be related to inflammatory reactions in the colon, possibly in connection with the initiation of an immunological response in sites distant from the parasite larvae.     

Parasite population dynamics in pigs infected with Trichuris suis and Oesophagostomum dentatum

The aim of the present study was to investigate the population dynamics and potential interactions between Trichuris suis and Oesophagostomum dentatum in experimentally co-infected pigs, by quantification of parasite parameters such as egg excretion, worm recovery and worm location. Forty-eight helminth naïve pigs were allocated into four groups. Group O was inoculated with 20 O. dentatum L₃/kg/day and Group T with 10 T. suis eggs/kg/day. Group OT was inoculated with both 20 O. dentatum L₃/kg/day and 10 T. suis eggs/kg/day, while Group C was kept as an uninfected control group. All inoculations were trickle infections administered twice weekly and were continued until slaughter. Faecal samples were collected from the rectum of all pigs at day 0, and twice weekly from 2 to 9 weeks post first infection (wpi). Six pigs from each group were necropsied 5 wpi and the remaining 6 pigs from each group were necropsied 10 wpi. The faecal egg counts (FEC) and total worm burdens of O. dentatum were dramatically influenced by the presence of T. suis, with significantly lower mean FECs and worm burdens at 5 and 10 wpi compared to single infected pigs. Furthermore, in the presence of T. suis we found that O. dentatum was located more posteriorly in the gut. The changes in the Trichuris population were less prominent, but faecal egg counts, worm counts 5 wpi (57% recovered vs. 39%) and the proportion of infected animals at 10 wpi were higher in Group OT compared to Group T. The location of T. suis was unaffected by the presence of O. dentatum. These results indicate an antagonistic interaction between T. suis and O. dentatum which is dominated by T. suis.     

Immune and inflammatory responses in pigs infected with Trichuris suis and Oesophagostomum dentatum

The aim of the present study was to investigate parasite induced immune responses in pigs co-infected with Trichuris suis and Oesophagostomum dentatum as compared to mono-species infected pigs. T. suis is known to elicit a strong immune response leading to rapid expulsion, and a strong antagonistic effect on O. dentatum populations has been observed in co-infected pigs. Forty-eight helminth naïve pigs were allocated into 4 groups in a 2-factorial design. Two groups were trickle inoculated with either 10 T. suis eggs/kg/day (Group T) or 20 O. dentatum L3/kg/day (Group O). Group OT was infected with same levels of both T. suis and O. dentatum (Group OT) and Group C remained uninfected. In each group, six pigs were necropsied after 35 days and the remaining pigs after 71 days. Parasite E/S-antigen specific serum antibodies were quantified by an in-direct ELISA. qPCR was used to measure the expression of immune function related genes in the mucosa of proximal colon and the draining lymph node. Highly significant interactions were identified for O. dentatum specific IgG1 (p < 0.0001) and IgG2 (p < 0.0006) antibodies with a remarkable 2-fold higher antibody response in group OT pigs as compared to group O. These findings indicated that T. suis enhanced the antibody response against O. dentatum in Group OT. The gene expression data confirmed a strong Type 2 response to T. suis (e.g. marked increase in IL-13, ARG1 and CCL11) and clearly weaker in amplitude and/or delayed onset response to O. dentatum in the single infected group. Interactions were found between the two nematodes with regard to several cytokines, e.g. the increase in IL-13 observed in Group T was absent in Group OT (p = 0.06, proximal colon mucosa, 35 and 71 p.i.). Some of these immune response-related interactions may support, or even partially explain, the observed interactions between the two worm populations in co-infected pigs.     

Villous atrophy in pigs orally infected with Salmonella cholerae-suis.

The macroscopic and microscopic appearance of villous atrophy in weaned pigs orally infected with Salmonella cholerae-suis is described. The other intestinal diseases of pigs in which villous atrophy occurs are discussed.     

Enrichment for isolating Salmonella Choleraesuis and other Salmonella spp. from pigs

The growth of Salmonella Choleraesuis was examined in Rappaport Vassiliadis broth (RV) and Hajna-tetrathionate broth (HTT) at 37 and 42 degrees C. As the enrichment in RV at 37 degrees C was satisfactory for isolating S. Choleraesuis, we used this enrichment for isolation from the samples collected from 15 asymptomatic pigs reared on a S. Choleraesuis contaminated farm. S. Choleraesuis was frequently isolated from six pigs (40.0%) under field conditions. The isolation of other Salmonella serovars than S. Choleraesuis was attempted by using both RV enrichment at 37 degrees C and HTT enrichment at 42 degrees C. Salmonella organisms were isolated from 156 (44.8%) of 348 fecal samples and more frequently with HTT at 42 degrees C (43.4%) than with RV at 37 degrees C (20.9%). If other serovars in addition to S. Choleraesuis are to be surveyed, HTT enrichment should be used in combination with RV enrichment.     

猪食道口线虫Cold-SSCP鉴别方法的建立

食道口线虫病是由食道口科食道口属(Oe-sophagostomum)的多种线虫寄生于动物肠道引起的,严重感染时可引起结肠炎。目前,此病在世界各国及我国各地牛、羊、猪中普遍存在,给畜牧业生产造成较大经济损失,而且国外也有食道口线虫感染人的报道[1]。因此,对食道口线虫的研究,除对畜牧     

The effect of continuous in-feed medication with thiophanate on Ascaris suum and Oesophagostomum spp. egg output in young pigs

The full potential of anthelmintics now available for single dose treatment is not achieved because the devising system for worm control in piglets/weaners is not efficiently applicable in practice. Therefore an in-feed medication programme for growing young pigs, allowing only one feed lot to be handled by the farmer, was tested in two studies. Study A Feed containing 0.0225% thiophanate was continuously fed almost ad lib. to piglets from birth right up to about 25 kg body weight when ready for fattening. This control measure effectively prevented A. suum and Oesophagostomum from becoming established during the whole pre-fattening period, thus allowing "worm-free" weaners to be produced. -33% of animals receiving unmedicated feed harboured mature Oesophagostomum already at an age of 63 days when first examined. Three out of 97 unmedicated pigs were then A. suum egg-count positive. At the same time all medicated pigs, except one with a low Oesophagostomum egg output, were egg-count negative. All medicated were still egg-count negative at 23-29 days after the withdrawal of the feed. About 30% of unmedicated pigs were then shedding eggs of A. suum and Oesophagostomum respectively. At 45-49 days after the withdrawal of the medicated feed 8% of previously medicated pigs and 43% of unmedicated pigs were A. suum egg-count positive. The corresponding figures for Oesophagostomum egg-count positive pigs were 6% and 40% respectively. The acquisition of worm infections by previously medicated pigs most probably was made in the fattening unit after the withdrawal of the thiophanate medicated feed. Study B In this study it was further substantiated that in-feed medication of pigs with thiophanate prevents A. suum from becoming established. All treated pigs were A. suum egg-count negative at Day 43 and 46 after the withdrawal of the medicated feed whereas about 62% of untreated control pigs were shedding A. suum eggs at the same time. This finding justify the proposal that the in-feed medication performed prevented larval migration. Furthermore it was shown that the in-feed medication must proceed right up to the transfer of piglets to the fattening unit in order to achieve its full potential. Farrowing pens may be heavily contaminated with infective Oesophagostomum larvae at the end of the pre-fattening period resulting in sudden and heavy nodular worm infections after the withdrawal of the medicated feed.     

Efficacy of piperazine dihydrochloride against Ascaris suum and Oesophagostomum species in naturally infected pigs

A controlled trial was performed to evaluate the efficacy of piperazine dihydrochloride in a new granular formulation (Ascarex D) against naturally occurring infections with Ascaris suum, Oesophagostomum dentatum and O quadrispinulatum. Treatment effects were estimated on the basis of parasites recoverable from the intestinal contents. Given orally at 200 mg per kg body weight the compound showed an efficacy of 99 to 100 per cent against A suum and the nodular worms. Egg excretion of the respective species was reduced by 98 per cent and 100 per cent six days after treatment. No adverse reactions were observed after the treatment.     

The in vitro motility response to various anthelmintics of third-stage larvae of Oesophagostomum spp. from pigs

The in vitro activities of thiabendazole, levamisole, pyrantel, morantel and ivermectin against Oesophagostomum spp., the nodular worm of pigs, were determined and compared. The study was carried out using isolates of O. dentatum and O. quadrispinulatum, which had been defined in vivo. Infective larvae were exposed to the anthelmintics for 24 h and then placed in a micromotility meter. All the treatments significantly reduced the motility of the ensheathed L₃ larvae, but the micromotility meter was not able to differentiate between anthelmintic resistant and anthelmintic susceptible isolates.     

Evaluation of vaccination with a commercial subunit vaccine on shedding of Salmonella enterica in subclinically infected dairy cows

OBJECTIVE: To estimate the efficacy of a commercially available Salmonella enterica subunit vaccine on the subclinical shedding of S enterica in dairy cattle. DESIGN: Randomized, controlled trial. ANIMALS: 175 mature cows on 2 dairy farms with a history of S enterica infection. PROCEDURES: 25% of the mature cows from each herd were systematically randomized to receive an S enterica subunit vaccine following label guidelines. The remaining 75% of cows in each herd served as nonvaccinated controls. Fecal samples were collected from all cows at the time of initial vaccination (day 0), booster vaccination (day 14), 2 weeks following the booster vaccination (day 28), and 10 weeks following the start of the trial (day 70). All samples were processed on the day of collection and cultured for S enterica. RESULTS: 651 fecal samples were obtained over the entire study period. Salmonella enterica was recovered from 46 (7.1%) of the samples. Shedding of S enterica was similar for vaccinated and nonvaccinated control cows on each of the collection dates. CONCLUSIONS AND CLINICAL RELEVANCE: The study revealed no evidence that extralabel vaccination with a commercial subunit S enterica vaccine reduced shedding of S enterica in subclinically infected dairy cows in these herds.     

Evaluation of the components of a commercial probiotic in gnotobiotic mice experimentally challenged with Salmonella enterica subsp. enterica ser. Typhimurium

Vitacanis((R)), a probiotic preparation containing a Lactobacillus acidophilus, an Enterococcus faecium and a Saccharomyces cerevisiae, has been developed for the prevention of intestinal disorders in dogs and cats. In the present study, these microorganisms were tested jointly or singly during experimental infection of gnotobiotic mice with Salmonella Typhimurium. Four experimental groups consisting of animals given probiotics jointly or singly and a control group consisting of germfree mice were used. The groups were treated with one or three of the microorganisms (experimental) or PBS (control) 10 days before intragastric challenge with a suspension containing about 10(2) cells of the bacterial pathogen. A higher survival (P<0.05) was observed in gnotobiotic mice given E. faecium (82%). All the animals in the other groups died after the challenge but the survival time was longer (P<0.05) for groups given all three of the microorganisms (7.4+/-2.4 days) or given only L. acidophilus (7.2+/-2.9 days) than for the control mice (4.4+/-1.1 days) and the mice that received S. cerevisiae (4.9+/-1.6 days) mice. The survival data agreed with the histopathological findings which showed more severe liver and intestinal lesions in control mice and in mice given Saccharomyces. In vitro antagonistic assays showed inhibition growth of E. faecium and S. Typhimurium around the colonies of L. acidophilus and for S. Typhimurium around the colonies of E. faecium. However, in vivo, S. Typhimurium became similarly established in the digestive tract of gnotobiotic mice at levels ranging from 10(8) to 10(10)CFU/g of feces and remained at these high levels until the animals died or were sacrificed. Among the three probiotic components of the commercial product Vitacanis((R)), E. faecium was the only one that provided protection against challenge with S. Typhimurium. Protection was not due to the reduction of the intestinal populations of the pathogenic bacteria.