James D. McKean

Salmonella enterica Infections in Market Swine with and without Transport and Holding

ABSTRACT The objective of this study was to compare, by using identical sample types, the Salmonella enterica prevalences and serovar diversities between pigs necropsied on the farm and those necropsied at the abattoir after transport and holding. We necropsied 567 market weight pigs (>70 kg) from six herds. Pigs were alternately assigned to be necropsied on the farm or at the abattoir. One-half of the group was sent in clean, disinfected trailers to slaughter at a commercial abattoir. After transport (mean distance, 169 km) and 2 to 3 h of holding in antemortem pens, these pigs were necropsied. The 50 pigs remaining on the farm were necropsied the following day. The same sample types and amounts were collected for S. enterica culture at both locations. Results show a sevenfold-higher (P < 0.001) S. enterica isolation rate from pigs necropsied at the abattoir (39.9%; 114 of 286) than from those necropsied on the farm (5.3%; 15 of 281). This difference was also observed for each individual herd. All sample types showed a significantly higher prevalence when comparing abattoir to on-farm collection, respectively: lymph nodes, 9.15 versus 3.6%; cecal contents, 13.6 versus 1.8%; 1 g of fecal matter, 25.2 versus 0.7%. Recovery of additional serovars at the abattoir suggests the pigs are receiving S. enterica from extra-farm sources. This study demonstrates that rapid infection during transport, and particularly during holding, is a major reason for increased S. enterica prevalence in swine. This finding identifies the holding pen as an important S. enterica control point in the pork production chain.

The effect of lairage on Salmonella isolation from market swine.

The objective of this paper was to evaluate the effect of lairage (holding >12 h during transport to slaughter) in clean facilities on Salmonella isolation from market swine. We tested 30 market-bound pigs (about 240 lb [110 kg]) on each of 10 occasions from an Iowa farrow-to-finish operation with about 600 sows. All pigs were slaughtered, and samples were collected at a large Midwest abattoir. On the farm, fecal samples were collected for culture of Salmonella. Pigs were alternately assigned to a lairage treatment (holding in a clean, disinfected facility at the National Animal Disease Center) group or a control group (remaining on the farm). After about 18 h, both groups were transported (about 137 km) to a large Midwest abattoir, commingled, and slaughtered. After slaughter, samples were collected for culture of Salmonella (feces from the distal colon, ileocecal lymph nodes, cecal contents, ventral thoracic lymph nodes, subiliac lymph nodes, and carcass swabs). Diaphragm sections were collected for serum ELISA. Salmonella enterica Derby was the only serotype isolated from farm fecal samples (3.4%, 10 of 290). Multiple serotypes (n = 17) were isolated from 71.8% (196 of 273) of the pigs when abattoir-collected samples were cultured: cecal contents (21.2%. 58 of 273), distal colon contents (52%, 142 of 273), and ileocecal lymph nodes (43.6%, 119 of 273). There were lower Salmonella isolation rates from the lairaged pigs (P < 0.05). The predominant serotype isolated at the abattoir varied by week of the study. This study suggests that pigs became internally contaminated with Salmonella after leaving the farm, possibly while in the abattoir holding pens, and that 18 h lairage, in clean facilities, does not increase shedding.

Preslaughter Holding Environment in Pork Plants Is Highly Contaminated with Salmonella enterica

ABSTRACT The objective of this study was to determine whether abattoir pens can provide a Salmonella enterica infection source during the 2 to 4 h of preharvest holding. Previous work has suggested that pigs may be getting infected, but little has been reported on the environmental contamination of abattoir holding pens. For 24 groups of pigs studied (∼150 animals/group) at two high-capacity abattoirs, six pooled fecal samples (n, 10 per pool) were collected from each transport trailer immediately after pigs were unloaded. Holding pens were sampled (one drinking water sample and six pooled floor samples consisting of swabs, residual liquid, and feces) prior to entry of study pigs for the routine holding period (∼2.5 h). After slaughter, cecal contents and ileocecal lymph nodes were collected, on the processing line, from 30 pigs in each studied group. All samples were cultured for the isolation and identification of S. enterica by primary enrichment in GN-Hajna and tetrathionate broths, secondary enrichment in Rappaport-Vassiliadis broth, and plating on brilliant green sulfa and xylose-lysine-tergitol-4 agars, followed by biochemical and serological identification. The study pens were highly contaminated with S. enterica; all holding pens sampled had at least one positive sample. Additionally, 33% (8 of 24) of drinking water samples were positive for S. enterica. All 24 groups of pigs had S. enterica-positive cecal contents and ileocecal lymph nodes, including those groups from transport trailers with no positive samples. From pigs, trailers, and pens, 586 isolates representing 36 different Salmonella serovars were isolated. Of the 353 isolates from pigs (109 from ileocecal lymph nodes plus 244 from cecal contents), 19% were identified as belonging to the same serovars as those isolated from the respective pens; 27% were identified as belonging to the same serovars as those isolated from the trailers. Sixteen percent of the unique serovars were isolated from both pigs and pens, suggesting that pens served as the infection source. This study demonstrates highly contaminated abattoir holding pens and watering sources. It also demonstrates that holding pens can serve as an infection source. This study identifies the abattoir holding pens as a significant hazard and a potential control point for Salmonella contamination in the preharvest pork production chain.

Feeding management practices and feed characteristics associated with Salmonella prevalence in live and slaughtered market-weight finisher swine: A systematic review and summation of evidence from 1950 to 2005

This review summarizes evidence for associations between Salmonella prevalence in market-weight swine and changes in feeding management practices or feed characteristics. A systematic review of the topic was conducted with the goal of minimizing the impact of bias on the review conclusions. Potential interventions included feed withdrawal from swine prior to slaughter, feed acidification, heat treatment of feed, pelletized feed versus mash, course versus fine grind, and wet versus dry feeds. In the reviewed literature, Salmonella prevalence was measured either by culture or by the presence of antibodies to Salmonella. The evidentiary value of studies was assessed, and studies that failed to meet predetermined standards were excluded. 7694 potentially relevant references were identified by an extensive literature search; however, 2623 references that were not published in English were excluded, because funds for translation were not available. Of the remaining references, only 277 were considered relevant to the review topic by two independent reviewers, and assessed for methodological quality. During quality assessment, 233 references were excluded because they failed to report design features that limit the introduction of bias or were conducted in a non-target population such as gnotobiotic, neonatal, nursery, or recently weaned pigs and sows. Forty-four publications passed the quality assessment conducted by 2 independent reviewers, but only 15 of the 44 publications reported studies that tested hypotheses associated with feeding management practices and feed characteristics and Salmonella prevalence in market-weight swine. The most common study design was cross-sectional (7/15). The included studies failed to provide strong evidence of an association between any of the potential interventions and Salmonella prevalence, due to the potential for confounding, and the failure to document temporal association between the intervention and Salmonella prevalence. The strongest evidence of an association was found for feed form, i.e. the use of non-pelleted may be potential interventions associated with reduced Salmonella prevalence. The uncertainty is primarily based on studies containing moderate to low evidentiary value or insufficient numbers of tested individuals, resulting in a low degree of confidence that results could be extrapolated to the target population. Therefore, the conclusion of the review is that there should be a low level of comfort among qualified scientists that the claimed association between non-pelleted feed and reduced Salmonella prevalence is scientifically valid. There is no strong evidence regarding associations between presence of Salmonella and the other feed characteristics examined.

Estimation of the Salmonella enterica prevalence in finishing swine.

The study objective was to evaluate three methods of Salmonella enterica prevalence estimation in swine herds (faecal culture, culture of abattoir-collected samples, and serum ELISA). From each of six swine herds, we necropsied approximately 100 finishing pigs (> 70 kg); one-half on farm and the other half at the abattoir, after transport and approximately 2.5 h holding. We collected the same samples for S. enterica culture at both locations (1 g faecal, 10 g caecal contents, ileocaecal lymph nodes, superficial inguinal lymph nodes, 25 g of gluteal muscle for serum ELISA). On farm, the 1 g faecal sample only detected 13.3% (2/15) of all positive pigs necropsied on farm. However, with abattoir and on-farm results combined, the faecal sample detected 57.4% (74/129) of positive pigs. Abattoir-collected samples provided prevalence estimates much higher than on-farm collected samples (39.9 vs. 5.3%; P < 0.001). This study shows that faecal samples have a low sensitivity for detecting infected pigs and that abattoir-collected samples overestimate the on-farm S. enterica prevalence. For most herds, serology overestimated the on-farm culture prevalence.

Detection of Listeria monocytogenes in pigs and pork

In this study, we surveyed hogs (n = 300) as well as pork products (ground pork and raw chitterlings) for Listeria monocytogenes. Pig specimens collected before (tonsil swabs) and after slaughter (tonsils, lymph nodes, carcass swabs, and rectal contents) were examined for L. monocytogenes by enrichment with conventional enrichment broths followed by subculturing to selective agar. A multiplex polymerase chain reaction assay targeting the highly conserved 16S rRNA gene of the Listeria species as well as the hlyA gene unique to L. monocytogenes was used to screen aliquots of the enrichment (method I) as well as to confirm presumptive Listeria colonies from Columbia agar with 0.05% glucose supplemented with polymyxin B-acriflavine-lithium chloride-ceftazidime-aesculin-mannitol (PALCAM; method II). Subculturing to PALCAM agar was the more sensitive of the two methods on the basis of the overall detection of Listeria. For hog tissues, method I detected L. monocytogenes (0.87% positive) and no other Listeria spp. in all samples (n = 1,849). In contrast, method II detected significantly more (P < 0.05) L. monocytogenes (2.38%) and Listeria spp. (0.38%) in these tissues. For small intestines (n = 300 raw chitterlings), L. monocytogenes was identified in 8.3% of enrichments with University of Vermont modified Listeria enrichment broth; plating to PALCAM slightly improved recovery (9%). Overall, ground pork samples (n = 340) harbored L. monocytogenes (45% positive) and other Listeria species (1.5% positive), as determined by method I. Subculturing to PALCAM significantly (P < 0.05) improved the detection of L. monocytogenes (50.2%) but not that of other Listeria species (1.7%). L. monocytogenes isolates (n = 243) were assigned to serotype 1 (53.5%), serotype 4 (25%), and serotypes other than 1 and 4 (21.4%).

The association between cleaning and disinfection of lairage pens and the prevalence of Salmonella enterica in swine at harvest

A series of four field trials were conducted to evaluate the ability of a cleaning and disinfection procedure in swine lairage pens to reduce the prevalence of Salmonella enterica in slaughtered pigs. A cleaning and disinfection procedure was applied to lairage pens at a large Midwest abattoir. Each trial consisted of a cleaned (alkaline chloride detergent) and disinfected (H2O2 plus peracetic acid sanitizer) pen (treated) and a control pen, each holding 90 to 95 pigs for 2 to 3 h before slaughter. Ileocecal lymph nodes, cecal contents, and rectal contents were collected from 45 pigs from each study pen at harvest and cultured for S. enterica. In all trials, cleaning and disinfection reduced the prevalence of S. enterica-positive floor swabs in the treated pen (P < 0.05). However, the postharvest prevalence of S. enterica-positive pigs varied between trials. In trial 1, there was no significant difference in the prevalence of S. enterica in pigs between treatment and control groups. In trials 2 and 3, the prevalence of S. enterica was higher in pigs from treated pens versus pigs from control pens (91% versus 40%, P < 0.0001, and 91% versus 24%, P < 0.0001, respectively). In trial 4, the prevalence of S. enterica was lower in pigs from treated pens compared with pigs from control pens (5% versus 42%, P < 0.0001). This study indicates that cleaning and disinfection effectively reduces the amount of culturable S. enterica in lairage pens, but the ability of cleaned and disinfected pens to reduce the prevalence of S. enterica in market-weight pigs remains inconclusive.

Impact of commercial preharvest transportation and holding on the prevalence of Salmonella enterica in cull sows.

The objective of this study was to examine the prevalence of Salmonella enterica in cull sows at various stages from the farm to the abattoir. Cull sows (n=181) were sampled over 10 weeks. Fecal samples (10 g each) were collected on the farm ca. 24 h before loading and at the live-hog market ca. 3 h before loading. Samples (ileocecal lymph nodes, cecal contents, feces from the transverse colon, ventral thoracic lymph nodes, subiliac lymph nodes, sponge swabs of the left and right carcass sections, and chopped meat) were collected at the abattoir. The percentages of positive fecal samples on the farm and at the live-hog market were 3% (5 of 181 samples) and 2% (3 of 181 samples), respectively. After transport from the live-hog market (10 h) and holding at the abattoir (6 h), 41% (74 of 180) of cull sows yielded S. enterica in one or more sampled tissues. The isolation rate for total cecal contents (33%; 60 of 180 samples) was significantly (P<0.05) higher than those for ileocecal lymph nodes (7%; 12 of 181 samples), feces (11%; 20 of 181 samples), and ventral thoracic and subiliac lymph nodes (2%; 4 of 181 samples). Before a 2% lactic acid carcass wash (lasting 8 to 9 s), 14% (25 of 180) of carcasses were positive, compared with 7% (12 of 179) after the wash (P<0.05). Two S. enterica serotypes, Derby and Infantis, were found on the farm and at the live-hog market. At the abattoir, 12 serotypes that had not previously been found on the farm or at the live-hog market were recovered. The results of this study demonstrate that transport and holding practices may contribute to an increase in S. enterica infection prior to slaughter to levels much higher than those found on the farm.

Effect of Short-Term Lairage on the Prevalence of Salmonella enterica in Cull Sows†

This study was designed to compare Salmonella enterica prevalence in sows held in a holding pen at the abattoir for approximately 2 h (hold sows) with sows slaughtered immediately after transport to the abattoir (no-hold sows). Cull sows (n = 160) were sampled from four sampling periods over 8 weeks (February to March 2002) at the abattoir. Sows originated from an integrated swine farm and were sent to a live-hog market and then to the slaughter facility. Before testing, sows entered the abattoir pen and four 100-cm2 four-ply gauze squares were placed randomly on the pen floor for S. enterica culture. Sows were alternatively assigned to the hold or no-hold group. Samples collected from sows during slaughter were ileocecal lymph node, cecal contents, transverse colon contents, subiliac lymph node, sponge swabs of the left and right carcass section (300 cm2), and chopped meat. Overall, S. enterica was isolated from 44% (35 of 80) of the no-hold sows, which was significantly less (P < 0.05) than 59% (47 of 80) of the held sows. Also, no-hold sows had a lower cecal content prevalence (39%, 31 of 80) compared with that (55%, 44 of 80) of held sows (P < 0.05). S. enterica serovars isolated from no-hold sows were Brandenburg (n = 16), Derby (n = 12), Hadar (n = 8), Infantis (n = 6), Johannesburg (n = 3), 6,7:z10-monophasic (n = 3), and Typhimurium (n = 1). S. enterica serovars isolated from held sows (n = 61 isolates) were Derby (n = 19), 6,7: z10-monophasic (n = 15), Brandenburg (n = 10), Infantis (n = 6), Hadar (n = 5), Johannesburg (n = 4), and Tennessee (n = 2). Serovars recovered from the pen were Reading (n = 6), Derby (n = 4), Uganda (n = 2), and Manhattan (n = 2). Results of this study suggest that holding pens contribute to increased S. enterica carriage in cull sows. Abattoir holding pens might be an important control point for S. enterica in the ground pork production chain.

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.