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1.
OBJECTIVE: To determine whether stress associated with transportation or feed withdrawal increased fecal shedding of Salmonella Typhimurium among pigs experimentally infected with the organism. ANIMALS: 86 healthy pigs. PROCEDURE: Pigs were challenge exposed with Salmonella Typhimurium at 4 weeks old and reared conventionally. When pigs reached market weight, they were assigned to groups and subjected to various combinations of transportation and feed withdrawal. Ileocecal contents were collected after slaughter and tested for Salmonella Typhimurium. RESULTS: Salmonella Typhimurium was not detected in feces collected from pigs just prior to slaughter. When feed was withheld for 24 hours prior to slaughter, the proportion of transported pigs with Salmonella Typhimurium in ileocecal contents at the time of slaughter was not significantly different from the proportion of nontransported pigs. However, when feed was not withheld prior to slaughter, the proportion of transported pigs with Salmonella Typhimurium in ileocecal contents at the time of slaughter was significantly higher than the proportion of nontransported pigs. CONCLUSIONS AND CLINICAL RELEVANCE: When carrier pigs remained on feed, transportation stress increased the proportion positive for Salmonella sp. On the basis of results reported here, it is suggested that producers withhold feed from pigs for 24 hours prior to transportation to a slaughter plant.  相似文献   

2.
In this study, different strategies to reduce carriage of Salmonella spp. in pigs were evaluated. Probiotics, prebiotics, vaccination, and acidification of drinking water were assessed as means of reducing Salmonella. Acidification of water, use of egg yolk-specific immunoglobulins, and vaccination with an endotoxin vaccine did not reduce Salmonella excretion in experimentally infected pigs. A reduction of Salmonella in the colonization of mesenteric lymph nodes was observed with the use of bambermycins and a live attenuated vaccine. A reduction in the shedding of S. Typhimurium was also observed after supplementation with fructooligosaccharides in drinking water. The use of probiotics and prebiotics appeared to change the pig fecal bacterial flora as indicated by Gram staining of smears from rectal swabs.  相似文献   

3.
The present study was conducted to study the impact of the virulence factors invC and sseD of the two type III secretion systems of Salmonella enterica serovar Typhimurium (S. Typhimurium) on the pathogenesis of the porcine S. Typhimurium DT104 infection. For this purpose, two S. Typhimurium mutant strains with a disrupted invC gene of the Salmonella pathogenicity island 1 or with a disrupted sseD gene of the Salmonella pathogenicity island 2 have been studied in experimental infection of pigs. Twenty-two 7-week-old male hybrid pigs were either infected with one of the mutants or the wild-type S. Typhimurium DT104 strain. Each group was examined for clinical signs, Salmonella shedding rate and the specific antibody response. Survival and replication were evaluated by qualitative and quantitative determination of the colonization rate. The humoral and cellular immune responses were examined using isotype-specific ELISA and quantitative real-time PCR of IL-2, IL-4, IL-10, IL-12 and IFN-gamma. The results proved that both mutants had a lower virulence (with marked differences between both mutants) than the wild-type and that both virulence factors have importance in porcine salmonellosis. Only pigs infected with the wild-type S. Typhimurium DT104 exhibited typical clinical symptoms of salmonellosis like anorexia, vomiting, disturbed demeanour, fever and diarrhoea. Deletion of the invC gene resulted in a significantly reduced colonization rate. Interestingly, the mRNA expression of both type-1 and type-2 cytokines were significantly decreased in pigs infected with either the invC-mutant and the sseD-mutant strain.  相似文献   

4.
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5.
Foodborne salmonellosis is one of the most important bacterial zoonotic diseases worldwide. Salmonella Typhimurium is the serovar most frequently isolated from persistently infected slaughter pigs in Europe. Salmonella Typhimurium pathogenesis is host species specific. In addition, differences in in vitro behaviour of Salmonella Typhimurium strains have also been described, which may be reflected by a different course of infection within a host species. We compared the course of a Salmonella Typhimurium infection in pigs, using two Salmonella Typhimurium strains that were able to interfere with MHC II expression on porcine macrophages to a different extent in vitro. After experimental inoculation, blood and faecal samples from all pigs were collected at regular time points. At 40 days post inoculation (pi), animals were euthanized and tissue samples were bacteriologically analysed. The proportion of serologically positive piglets at 33 days pi was significantly higher in pigs that were inoculated with the strain that did not downregulate MHC II expression in vitro. Furthermore, this strain was less frequently shed and isolated in lower numbers from tonsils and ileocaecal lymph nodes than the strain that was able to markedly downregulate MHC II expression in vitro. We thus found that the delayed onset of seroconversion after oral inoculation of piglets with a particular Salmonella Typhimurium strain coincided with higher faecal shedding and increased persistence. Strain specific differences in Salmonella pathogenesis might thus have repercussions on the serological detection of Salmonella Typhimurium infections in pigs.  相似文献   

6.
In this study, we investigated the effect of adding organic acids to the drinking water of finishing pigs 2 weeks prior to slaughter on the shedding and prevalence rate of Salmonella at slaughter. Approximately 600 animals from four Belgian pig herds infected with Salmonella were included. At two herds, the study was conducted twice. Before the start of the study, overshoes were taken at the different herds. Two weeks prior to the expected slaughter date, the pigs were randomly divided into two groups (treatment and control group) each containing on average 50 animals within each herd. The treatment group received from this day onwards acidified drinking water (pH = 3.6-4.0), the control group received non-treated water (pH = 7.8-8.5). All other housing, feeding and management factors were identical in both groups. At the slaughterhouse, 10 pigs of each group (20 pigs for each group of study group 6) were randomly selected and sampled (blood, contents of ileum and rectum, mesenteric lymph nodes and carcass swabs). All samples were immediately transported to the laboratory and submitted to Salmonella isolation. Salmonella was isolated out of 11.9% (66/554) of the samples taken at the slaughterhouse, with the highest frequency found in the content of the ileum (18.7%), followed by 17.8% in the lymph nodes, 7.2% in the content of the rectum and 3.6% in the carcass swabs. The results did not reveal a significant difference between the treatment and control groups for the different slaughterhouse samples. The study documented that the investigated control strategy namely, the strategic application of organic acids during the last 2 weeks prior to slaughter was insufficient to decrease Salmonella shedding and contamination shortly before and during slaughter.  相似文献   

7.
Specified pathogen-free cats were naturally infected with FCoV or experimentally infected with FCoV type I. Seroconversion was determined and the course of infection was monitored by measuring the FCoV loads in faeces, whole blood, plasma and/or monocytes. Tissue samples collected at necropsy were examined for viral load and histopathological changes. Experimentally infected animals started shedding virus as soon as 2 days after infection. They generally displayed the highest viral loads in colon, ileum and mesenteric lymph nodes. Seroconversion occurred 3-4 weeks post infection. Naturally infected cats were positive for FCoV antibodies and monocyte-associated FCoV viraemia prior to death. At necropsy, most animals tested positive for viral shedding and FCoV RNA was found in spleen, mesenteric lymph nodes and bone marrow. Both experimentally and naturally infected cats remained clinically healthy. Pathological findings were restricted to generalized lymphatic hyperplasia. These findings demonstrate the presence of systemic FCoV infection with high viral loads in the absence of clinical and pathological signs.  相似文献   

8.
Salmonella Typhimurium infections in pigs are a major source of human foodborne salmonellosis. To reduce the number of infected pigs, acidification of feed or drinking water is a common practice. The aim of the present study was to determine whether some frequently used short- (SCFA) and medium-chain fatty acids (MCFA) are able to alter virulence gene expression and to decrease Salmonella Typhimurium colonization and shedding in pigs using well established and controlled in vitro and in vivo assays. Minimal inhibitory concentrations (MIC) of 4 SCFA (formic acid, acetic acid, propionic acid and butyric acid) and 2 MCFA (caproic and caprylic acid) were determined using 54 porcine Salmonella Typhimurium field strains. MIC values increased at increasing pH-values and were two to eight times lower for MCFA than for SCFA. Expression of virulence gene fimA was significantly lower when bacteria were grown in LB-broth supplemented with sub-MIC concentrations of caproic or caprylic acid (2 mM). Expression of hilA and invasion in porcine intestinal epithelial cells was significantly lower when bacteria were grown in LB-broth containing sub-MIC concentrations of butyric acid or propionic acid (10 mM) and caproic or caprylic acid (2 mM). When given as feed supplement to pigs experimentally infected with Salmonella Typhimurium, coated butyric acid decreased the levels of faecal shedding and intestinal colonization, but had no influence on the colonization of tonsils, spleen and liver. Uncoated fatty acids, however, did not influence fecal shedding, intestinal or tonsillar colonization in pigs. In conclusion, supplementing feed with certain coated fatty acids, such as butyric acid, may help to reduce the Salmonella load in pigs.  相似文献   

9.
The implementation of Salmonella control programs in the pork production chain demands rapid and cost-effective methods to assess the prevalence of infection in pig herds. The objective of the present study was to develop an in-house enzyme-linked immunosorbent assay (ELISA) based on S. Typhimurium lipopolysaccharides (LPS) to measure the prevalence of infection caused by Salmonella in swine herds. Coating antigen was produced by phenol extraction of S. Typhimurium culture. After standardization of ELISA test conditions, the assay was validated by testing serum samples on different animal categories: pigs orally inoculated with S. Typhimurium and sentinel animals in contact with them, naturally infected animals, colostrum-deprived piglets, and bacterin-immunized pigs. Seroconversion was observed in inoculated pigs (7 days postinfection [DPI]) and in the sentinels (21 DPI). Nonspecific reactions were not detected in the sera of colostrum-deprived animals. Serum samples from animals immunized with Salmonella Agona, Salmonella Derby, Salmonella Panama, and Salmonella Bredeney bacterins showed marked cross-reaction with the LPS from the serovar Typhimurium. Moreover, positive results obtained with the in-house ELISA were associated with Salmonella isolation in 75 infected pig herds. Comparisons with 2 commercial kits showed a linear correlation coefficient of 0.847 between the in-house ELISA and kit A and 0.922 with kit B but a low agreement in the qualitative results. In conclusion, the newly developed in-house ELISA based on S. Typhimurium LPS can be a useful tool to determine the intensity of Salmonella sp. infection in swine herds.  相似文献   

10.
The objective of these experiments was to evaluate the possibility of swine becoming infected with Salmonella Typhimurium after a short time interval in a contaminated environment. Two experiments were conducted. Experiment 1 consisted of five trials with eight market weight swine. Pigs were necropsied at 2 (n = 10), 3 (n = 10) and 6 (n = 5) hours after continuous exposure to an environment contaminated with feces shed by swine intranasally inoculated with nalidixic acid-resistant Salmonella Typhimurium (chi 4232). In Experiment 2, pigs were necropsied after 30 minutes (n = 6), 60 minutes (n = 6), 2 hours (n = 6), and 6 hours (n = 3). In addition, control animals with no exposure were also necropsied in both experiments. At necropsy, the superficial inguinal, ileocecal, and mandibular lymph nodes, as well as cecal contents, distal ileum portion, and feces were evaluated. All samples were cultured for the presence of the nalidixic acid-resistant Salmonella. Feces deposited on the floor by intranasally inoculated swine were mixed with water to form slurry with a resulting load of 10(3)-10(5) Salmonella Typhimurium CFU per gram. In Experiment 1, 80% percent of animals with a 2-hour, 60% of animals with a 3-hour, and 100% of animals with a 6-hour exposure to this slurry had at least one sample test positive for the marked Salmonella Typhimurium strain. In Experiment 2, 50% of the 30 minute, 50% of the 60 minute, and 33% of the 2-hour exposed pigs had at least one sample test positive. These experiments show that market swine can become infected during routine resting or holding periods when exposed to relatively low levels (10(3) CFU) of Salmonella in the simulated pre-slaughter environment, and that exposure times as short as 30 minutes are sufficient to produce contaminated gastrointestinal tracts. They also demonstrate the high risk of holding pigs longer than six hours. Intervention at this step in the swine production process may have a significant impact on the safety of pork products.  相似文献   

11.
A clinical field trial was performed to determine the effectiveness of an autogenous Salmonella Typhimurium bacterin compared with a commercial live S. Choleraesuis vaccine in pigs. The association between Salmonella shedding and weight gain was also investigated. Nine cohorts of weaned pigs, (330 to 350 pigs per cohort), were randomly assigned to 1 of 3 treatment groups (injection with S. Typhimurium bacterin, vaccination via water with S. Choleraesuis vaccine, or a control group receiving no vaccine). In each cohort, the average daily gain was calculated for a selected pen throughout the production stage. Pen (pooled) fecal samples were collected bi-weekly and cultured. The odds of Salmonella shedding in both vaccinated groups was higher than in the control group (P < 0.05). The prevalence of Salmonella shedding declined overall as pigs aged (P = 0.04). However, the control pigs showed the smallest decrease in Salmonella shedding over the entire production stage, while prevalence of Salmonella shedding in the vaccinated groups decreased twice as much as the control group over the entire production stage. Salmonella Typhimurium var. Copenhagen DT104, S. Cerro, and S. Agona, which had been isolated on the study farm previously, were recovered from pigs in this study. Shedding of S. Typhimurium var. Copenhagen decreased over time in both vaccine treatment groups. On the other hand, S. Cerro shedding rate was lower in the control pigs compared with vaccinated pigs and S. Agona could be recovered only from the samples collected from S. Choleraesuis vaccinated pigs. The pigs from pens with a higher Salmonella recovery rate experienced slower growth compared with pigs from pens where Salmonella was not isolated. This latter finding indicates that there might be an economic incentive for producers to try to control endemic salmonellosis if effective programs could be developed.  相似文献   

12.
Although poultry is recognized as the major source of food-poisoning caused by Salmonella, pork also contributes to human infections. This study was therefore undertaken in order to develop a reliable serological method for the evaluation of the Salmonella status of piglets. A complete ELISA was performed using lipopolysaccharides of Salmonella Typhimurium, Anatum, Hadar and Infantis because these serovars were representative of the serogroups isolated from 30 contaminated fattening farms. S. Enteritidis was also added because of its importance in human infection and to include the O:9 antigen. This method potentially detects 100% of infected pigs. A significant correlation was found between this serological method and the bacteriological data from mesenteric lymph nodes (p = 0.01). In addition, both sensitivity and specificity were high (97% and 94% respectively). The ELISA test was therefore used in a cross-sectional study on 4 farms to evaluate when pigs became contaminated: seropositive pigs were only found for the 20 week old finishing pigs. The antibody response to Salmonella in piglets was also investigated: maternal antibodies persisted until 7 weeks of age and post-Salmonella contamination seroconversion was detected from 8 weeks of age onwards.  相似文献   

13.
A model for experimental and natural infection of early weaned pigs with Salmonella choleraesuis, the aetiologic agent of swine paratyphoid, has been developed. An oral dose of 108 colony forming units (cfu) of S choleraesuis caused 100 per cent infection of 10 pigs inoculated, as indicated by recovery of the challenge organism from ileocolic lymph nodes collected at necropsy seven days post challenge. Seven of the pigs were observed shedding S choleraesuis at least once post S choleraesuis challenge. The cumulative incidence of shedding was 30 per cent and was sufficient to infect four of 10 pigs exposed naturally. Oral challenges with less than 108 cfu S choleraesuis were less effective in infecting early weaned pigs and did not result in natural transmission.  相似文献   

14.
The prevalence of Salmonella spp. infection was determined in 67 free-range pig herds in southern Spain. Microbiological assessment was performed on ileocolic lymph nodes collected at slaughter according to ISO 6579:2002 procedures. Overall, 33% of herds were infected and the prevalence of infection was 5.3%. Salmonella spp. serovars most frequently isolated were Anatum and Typhimurium, although uncommon serovars such as Hessarek and Mikawasima were also detected. Isolates were tested against 16 antimicrobial agents and exhibited resistance to streptomycin (46%), tetracycline (30%), sulphonamides (25%) and ampicillin (23%) by the break-point method. Multi-drug resistance, defined as resistance to ≥ 4 antimicrobials, was 36%.  相似文献   

15.
OBJECTIVE: To investigate the effect of an Escherichia that produced microcin 24 (Mcc24) on shedding of of Salmonella enterica serotypeTyphimurium in swine and evaluate evidence of in vivo activation of the Mcc24-mediated, multiple-antibiotic resistance (mar) operon. ANIMALS: 36 crossbred weaned pigs. PROCEDURE: 24 pigs were allocated to 2 groups (12 pigs/group). Pigs in 1 group received daily oral administration of an Mcc24-producing E coli, whereas the other group received a non-Mcc24-producing E coli. All pigs were challenge exposed with Salmonella Typhimurium chi4232. A third group of 6 pigs received Mcc24-producing E coli and was challenge exposed with an Mcc24-sensitive, marA-deleted strain of Salmonella Typhimurium 4232. After challenge exposure, fecal samples from all pigs were cultured to detect shedding of Salmonella Typhimurium and Salmonella Typhimurium isolates were screened for resistance to ciprofloxacin. Fecal samples were collected throughout the study, and tissue samples were collected during necropsy. RESULTS: Differences in shedding of Salmonella Typhimurium were not detected between groups receiving Mcc24-producing or non-Mcc24-producing E coli. No significant differences were found in quantitative analysis between groups receiving Mcc24-producing and non-Mcc24-producing E coli. Evidence of mar activation was not detected. CONCLUSIONS AND CLINICAL RELEVANCE: Microcin-producing E coli did not exert an effect on shedding of SalmonellaTyphimurium or mar activation in pigs. It may be difficult or impractical to create the conditions required for Mcc24 to be an effective part of a food safety intervention to reduce shedding of Salmonella Typhimurium.  相似文献   

16.
Worldwide, the use of antimicrobials in food production has been associated with drug resistance in foodborne pathogens such as Salmonella. However, little is known about the efficaciousness of fluorequinolone treatment on Salmonella Typhimurium T104 infections in pig breeding herds. A combined eradication procedure with enrofloxacin application on sows and piglets, feeding of encapsulated organic acids to sows, disinfection with peracetic acid, separation of the growers from the sows and serological discrimination using a new whole-cell-based enzyme-linked immnosorbent assay (ELISA) was evaluated for the suitability to eradicate and to control endemic S. Typhimurium DT104 infections in a closed herd. Thirty-seven sows and their piglets were treated everyday from day 14 ante partum until the day of weaning. Eighteen sows and their piglets served as controls. From the first day of life until day 168 after birth, faecal samples (n = 1671) of all piglets were analysed for Salmonella shedding. In parallel, systemic antibody responses were monitored by whole-cell-based isotype-specific ELISA systems. From birth to weaning the prevalence in both groups was between 2% and 9%. After weaning, intermittent shedding could be observed in both groups, and salmonellae could be found in up to 7.7% of the faecal samples. As a result, a dramatic increase in Salmonella-infected growers was observed, as of day 115 after birth, 47.4% of the animals of the treated group were tested positive for S. Typhimurium. Our results indicate that despite long-term antibiosis treatment and optimized hygiene measures, shedding of S. Typhimurium by the sows and the subsequent infection of their offspring could not be effectively prevented. Although it could be not shown that elimination of S. Typhimurium DT104 infection was achieved, the disinfection procedures described and the diagnostic test used are effective instruments to decrease the Salmonella load and to identify individual infected animals. Both of these are important factors for an improved consumer protection.  相似文献   

17.
Clinical salmonellosis associated with Salmonella is increasingly reported in finishing swine. Since S. Typhimurium is often associated with these episodes and given that this serotype is among the most often reported in humans, we were interested to determine if various tissues and carcasses from animals coming from herds that were clinically affected were more likely to be contaminated by Salmonella compared to carcasses from animals raised in herds without any history of salmonellosis. Carcasses from animals from affected herds were significantly more contaminated by Salmonella while showing increased titers in antibodies directed against this bacterium. At the opposite, caecal contents and mesenteric lymph nodes from both groups of animals were similarly contaminated by Salmonella. In the second part of the study, we studied the persistence of the bacterium in various tissues after an experimental infection with S. Typhimurium. We found that, after the infection, Salmonella persisted for as many as 7 d in many extraintestinal tissues, while it was present in the feces of infected animals for all 14 d of the experiment. These findings indicated that carcasses from animals that experienced salmonellosis during their growth phase are more likely to be contaminated by this bacterium and that precautions must be taken in order to ensure that clinically affected animals should be kept on the farm for at least 7 d before being shipped for slaughter.  相似文献   

18.
Salmonella enterica serovar Typhimurium is an animal and zoonotic pathogen of worldwide importance. In pigs, transport and social stress are associated with reactivation and spread of Salmonella Typhimurium infection. The stress-related catecholamine norepinephrine (NE) has been reported to activate growth and virulence factor expression in Salmonella; however the extent to which NE contributes to stress-associated salmonellosis is unclear. We studied the impact of releasing NE from endogenous stores during Salmonella Typhimurium infection of pigs by administration of 6-hydroxydopamine (6-OHDA), which selectively destroys noradrenergic nerve terminals. Treatment of pigs with 6-OHDA 7 or 16 days post-oral inoculation with Salmonella Typhimurium produced elevated plasma NE levels and transiently, but significantly, increased faecal excretion of the challenge strain. Oral administration of NE to Salmonella Typhimurium-infected pigs also transiently and significantly increased shedding; however pre-culture of the bacteria with NE did not alter the outcome of infection. Salmonella has been proposed to sense and respond to NE via a homologue of the adrenergic sensor kinase QseC. A ΔqseC mutant of Salmonella Typhimurium was consistently excreted in lower numbers than the parent strain post-oral inoculation of pigs, though not significantly so. 6-OHDA treatment of pigs infected with the ΔqseC mutant also increased faecal excretion of the mutant strain, albeit to a lesser extent than observed upon 6-OHDA treatment of pigs infected with the parent strain. Our data support the notion that stress-related catecholamines modulate the interaction of enteric bacterial pathogens with their hosts.  相似文献   

19.
OBJECTIVE: To evaluate the possibility of swine becoming infected with Salmonella Typhimurium when housed for 2 to 6 hours in an environment contaminated with Salmonella, similar to a lairage situation prior to slaughter. ANIMALS: 40 crossbred market pigs with an approximate body weight of 92 kg. PROCEDURE: Five trials were conducted (8 pigs/trial) in simulated lairage conditions. Superficial inguinal, ileocecal, and mandibular lymph nodes, cecal contents, distal portion of the ileum, and fecal samples were obtained from each pig after 2 (n = 10), 3 (10), and 6 (5) hours of exposure to an environment contaminated with feces defecated by 10 pigs intranasally inoculated with nalidixic acid-resistant Salmonella Typhimurium (chi4232). In addition, 5 control pigs that were not exposed were also evaluated in the same manner. RESULTS: Feces deposited on the floor by intranasally inoculated swine were mixed with water to form slurry with a resulting load of approximately 10(3) colony-forming units of Salmonella Typhimurium/g of material. Eight of 10, 6 of 10, and 6 of 6 pigs exposed to the slurry for 2, 3, or 6 hours, respectively, had positive results for at least 1 sample when tested for the specific strain of Salmonella Typhimurium. CONCLUSIONS AND CLINICAL RELEVANCE: Pigs can become infected during routine resting or holding periods during marketing when exposed to relatively low amounts of Salmonella organisms in the preslaughter environment. Intervention at this step of the production process may have a major impact on the safety of pork products.  相似文献   

20.
Two trials were carried out to determine the effect of feed acidification upon Salmonella carriage in market-age pigs. In the first trial, the administration for the last 14 weeks of the fattening period of a commercial pelleted feed added with 0.6% lactic acid plus 0.6% formic acid (Lac-Formic-1.2) was compared to an unacidified standard diet (STD). A second experiment was carried out in two herds of growing pigs (Herd I, 3000 pigs; Herd II, 900 pigs) in which three different diets were assayed during the last 8-9 weeks of the fattening period: a diet containing 0.8% formic acid (Formic-0.8), a diet containing 0.4% lactic acid plus 0.4% formic acid (Lac-Formic-0.8) and a STD. In the first experiment, serological evolution of the infection was examined by ELISA and microbiological cultures (rectal samples and mesenteric lymph nodes) were also done. Feed intake by pen and the individual weight of the animals were also measured. In the second trial, blood, rectal samples and mesenteric lymph nodes were collected at slaughter in both herds (30 pigs per experimental group). In the first experiment, the acidified diet (Lac-Formic-1.2) reduced Salmonella carriers in mesenteric lymph nodes (Fisher's exact P < 0.01). In the second trial, Lac-Formic-0.8 diet significantly reduced Salmonella seroprevalence compared to the STD (P = 0.001) in both herds. Also Lac-Formic-0.8 and Formic-0.8 diets in Herd II showed a lower faecal excretion and Salmonella carriage in mesenteric lymph nodes than the STD (P < 0.05). Our results suggest that the administration of a combination of lactic and formic acids at the levels used in this study could be used to reduce Salmonella prevalence in finishing pigs.  相似文献   

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