Bovine and Pig Carcasses as a Source of Campylobacter in Poland: A Reservoir for Antimicrobial-Resistant Campylobacter coli.

Campylobacter is one of the most common causes of foodborne bacterial infections worldwide. Why poultry has been shown to be one of the most significant sources of these bacteria, ruminants, especially cattle, are also responsible for a high number of human Campylobacter jejuni, and to a lesser extent Campylobacter coli, infections. In this study, bovine and pig carcasses in Poland were investigated for the presence of Campylobacter and for their antimicrobial resistance. A total of 204 swabs from bovine carcasses and 355 swab samples from pig carcasses were tested during 2014-2018. Campylobacter was identified in 129 (36.3%) of the pig and in 11 (5.4%) of the bovine carcasses, respectively. The pig isolates were classified as C. coli (121; 34.1%) or C. jejuni (8; 2.3%), whereas the bovine Campylobacter were identified either as C. jejuni (8; 3.9% isolates) or C. coli (3; 1.5% strains). Resistance of the isolates (n = 140) to erythromycin, ciprofloxacin, nalidixic acid, streptomycin, and tetracycline revealed that the vast majority of C. coli was resistant to streptomycin (106 isolates; 85.5%), tetracycline (97; 78.2%), nalidixic acid (90; 72.6%), and ciprofloxacin (88; 71.0%). Among C. jejuni isolates (n = 16) the resistance rates to all antibiotics were lower than in C. coli, irrespective of the origin. A total of 74 of 121 (61.2%) C. coli isolates from the pig carcasses and one of three such isolates from the bovine samples were multiresistant. Most of the C. coli (64 isolates; 85.3%) had the ciprofloxacin+nalidixic acid+streptomycin+tetracycline resistance profile. The results suggest that pig and bovine carcasses may be an underestimated reservoir of Campylobacter, especially for C. coli in pigs. The high antimicrobial resistance rates of such strains to streptomycin, quinolones, and tetracyclines highlight the need for monitoring of these bacteria in such food and food products.

Acari community in association with delayed pig carrion decomposition.

Acari community structure and function associated with delayed pig carrion decomposition has not been examined. In this study, 18 swine carcasses were studied in central Texas, USA, during two consecutive summers (2013 and 2014). Samples of ca. 400 g soil were collected from beneath, aside, and 5 m away from each pig carcass over 180 days. Mites from soil samples were extracted using Berlese funnels and identified to order and family levels and classified according to ecological function. In total 1565 and 1740 mites were identified from the 2013 and 2014 soil samples, respectively. Significant differences were determined for mite community structure at order and family levels temporally on carrion (e.g., day 0 × day 14) regardless of treatments and between soil regions where mites were collected (e.g., soil beneath vs. soil 5 m away from carrion). However, no significant differences were found in mite community structure at the order level between pig carrion with and without delayed Diptera colonization (i.e., treatments). Analysis at the family level determined a significant difference across treatments for both summers. Ecological function of mites did not change significantly following the delayed decomposition of pig carcasses. However, detritivores and fungivores were significant indicator groups during the pig carrion decomposition process. Furthermore, 13 phoretic mite species associated with eight forensically important beetle species were documented. Data from this study indicated that the rate of nutrient flow into the soil impacted associated arthropod communities; however, detecting such shifts depends on the taxonomic resolution being applied.

Pigs vs people: the use of pigs as analogues for humans in forensic entomology and taphonomy research.

Most studies of decomposition in forensic entomology and taphonomy have used non-human cadavers. Following the recommendation of using domestic pig cadavers as analogues for humans in forensic entomology in the 1980s, pigs became the most frequently used model cadavers in forensic sciences. They have shaped our understanding of how large vertebrate cadavers decompose in, for example, various environments, seasons and after various ante- or postmortem cadaver modifications. They have also been used to demonstrate the feasibility of several new or well-established forensic techniques. The advent of outdoor human taphonomy facilities enabled experimental comparisons of decomposition between pig and human cadavers. Recent comparisons challenged the pig-as-analogue claim in entomology and taphonomy research. In this review, we discuss in a broad methodological context the advantages and disadvantages of pig and human cadavers for forensic research and rebut the critique of pigs as analogues for humans. We conclude that experiments using human cadaver analogues (i.e. pig carcasses) are easier to replicate and more practical for controlling confounding factors than studies based solely on humans and, therefore, are likely to remain our primary epistemic source of forensic knowledge for the immediate future. We supplement these considerations with new guidelines for model cadaver choice in forensic science research.

Identification and molecular characteristics of verotoxin-producing Escherichia coli (VTEC) from bovine and pig carcasses isolated in Poland during 2014-2018.

The presence of verotoxin-producing Escherichia coli (VTEC) on bovine (n = 330) and pig (n = 120) carcasses in Poland was investigated using the ISO/TS 13136 standard. A total of 115 (34.8%) and 37 (30.8%) cattle and pig samples were positive in real-time PCR, respectively. Isolation of the bacteria revealed that from bovine carcasses 37 (32.2%) VTEC were obtained whereas only 5 (13.5%) pig carcasses were positive for the stx gene. The VTEC were characterized using whole genome sequencing (WGS) and bovine isolates were classified into 25 serotypes with the most prevalent O113:H21 (5 strains) whereas pig strains belonged to 5 different serotypes which were not identified among cattle strains. The majority of bovine VTEC (35; 94.6% isolates) were positive for the stx2 gene, either alone or together with the stx1 gene. All strains isolated from pig carcasses resulted positive for the stx2 gene only. Only two isolates of bovine origin contained the eaeA intimin gene, together with the ehxA and lpfA markers. VTEC were highly molecularly diverse as shown by classification into 29 different MLST STs. The obtained results suggest that further studies related to cattle and pig carcasses are needed to assess the role of these sources for human VTEC infections.

Porcine carcasses as an underestimated source of antimicrobial resistant Campylobacter coli.

Introduction: Campylobacteriosis is the most common human foodborne bacterial infection worldwide and is caused by bacteria of the Camplylobacter genus. The main source of these bacteria is poultry, but other food-producing animals such as pigs are also responsible for human infections. An increasing number of strains with resistance to fluoroquinolones and other antimicrobials such as macrolides were recently noted. The aim of the study was to investigate Campylobacter contamination of porcine carcasses and determine the antimicrobial resistance of the obtained isolates. Material and Methods: A total of 534 swabs from carcasses of pigs slaughtered in Poland during 2019-2022 were tested for Campylobacter spp. Results: Campylobacter was detected in 164 (30.7%) carcasses; among them 149 (90.8%) were classified as C. coli and the remaining 15 (9.2%) samples were C. jejuni-positive. Because a low number of C. jejuni isolates were identified, only the C. coli isolates were subjected to antimicrobial resistance analysis. The majority of these isolates were resistant to streptomycin (94.0%), ciprofloxacin (65.8%) and tetracycline (65.1%). A total of 94 (63.1%) strains displayed antimicrobial multiresistance patterns and were mainly resistant to fluoroquinolones, aminoglycosides and tetracyclines (74; 49.7% of the isolates tested). Conclusion: The obtained results showed that pig carcasses may be contaminated with antimicrobial-resistant C. coli.

Systematic Review and Meta-Analysis of the Efficacy of Interventions Applied during Primary Processing to Reduce Microbial Contamination on Pig Carcasses.

Interventions from lairage to the chilling stage of the pig slaughter process are important to reduce microbial contamination of carcasses. The aim of this systematic review and meta-analysis was to assess the effectiveness of abattoir interventions in reducing aerobic colony count (ACC), Enterobacteriaceae, generic Escherichia coli, and Yersinia spp. on pig carcasses. The database searches spanned a 30 year period from 1990 to 2021. Following a structured, predefined protocol, 22 articles, which were judged as having a low risk of bias, were used for detailed data extraction and meta-analysis. The meta-analysis included data on lairage interventions for live pigs, standard processing procedures for pig carcasses, prechilling interventions, multiple carcass interventions, and carcass chilling. Risk ratios (RRs) for prevalence studies and mean log differences (MDs) for concentration outcomes were calculated using random effects models. The meta-analysis found that scalding under commercial abattoir conditions effectively reduced the prevalence of Enterobacteriaceae (RR: 0.05, 95% CI: 0.02 to 0.12, I2 = 87%) and ACC (MD: -2.84, 95% CI: -3.50 to -2.18, I2 = 99%) on pig carcasses. Similarly, significant reductions of these two groups of bacteria on carcasses were also found after singeing (RR: 0.25, 95% CI: 0.14 to 0.44, I2 = 90% and MD: -1.95, 95% CI: -2.40 to -1.50, I2 = 96%, respectively). Rectum sealing effectively reduces the prevalence of Y. enterocolitica on pig carcasses (RR: 0.60, 95% CI: 0.41 to 0.89, I2 = 0%). Under commercial abattoir conditions, hot water washing significantly reduced ACC (MD: -1.32, 95% CI: -1.93 to -0.71, I2 = 93%) and generic E. coli counts (MD: -1.23, 95% CI: -1.89 to -0.57, I2 = 61%) on pig carcasses. Conventional dry chilling reduced Enterobacteriaceae prevalence on pig carcasses (RR: 0.32, 95% CI: 0.21 to 0.48, I2 = 81%). Multiple carcass interventions significantly reduced Enterobacteriaceae prevalence (RR: 0.11, 95% CI: 0.05 to 0.23, I2 = 94%) and ACC on carcasses (MD: -2.85, 95% CI: -3.33 to -2.37, I2 = 97%). The results clearly show that standard processing procedures of scalding and singeing and the hazard-based intervention of hot water washing are effective in reducing indicator bacteria on pig carcasses. The prevalence of Y. enterocolitica on pig carcasses was effectively reduced by the standard procedure of rectum sealing; nevertheless, this was the only intervention for Yersinia investigated under commercial conditions. High heterogeneity among studies and trials investigating interventions and overall lack of large, controlled trials conducted under commercial conditions suggest that more in-depth research is needed.

Identification of the Source for Salmonella Contamination of Carcasses in a Large Pig Slaughterhouse.

To identify the major source of Salmonella contamination in a pig slaughterhouse, samples were collected from the clean and unclean area and Salmonella isolates were further typed. Carcasses entering the clean area showed a Salmonella contamination rate of 96.7% in the oral cavity and 55.0% in the rectum content samples. Evisceration seemed not to be critical as the contamination rate of the carcasses was similar before (16.7%) and after (18.3%) this slaughter step. In the unclean area, a limited number of oral cavity samples were positive after bleeding, while a dramatic increase of positives was observed after dehairing. Salmonella was detected in up to 0.01 mL of the recycled water collected from the dehairing machine. Genotyping of Salmonella isolates showed that similar pulsotypes were present in the oral cavity and recycled water. Based on these observations it can be concluded that the recycled water used in the dehairing machine was the major source for the carcass contamination in this slaughterhouse.

Seasonal and spatial occurrence of zoonotic Salmonella serotypes in griffon vultures at farmland environments: Implications in pathogen pollution and ecosystem services and disservices.

Livestock production is a large source of microbial, pharmaceutical, and antimicrobial pollution worldwide. Vultures are one group of birds with particularly high exposure to food-borne pathogens due to frequent consumption of infected livestock carcasses. The potential origin and spatial-temporal shedding patterns of livestock-adapted Salmonella serotypes of zoonotic importance were evaluated in adult and nestling Griffon vultures (Gyps fulvus). We specifically assessed the exposure source and subsequent elimination of Salmonella-infected carcasses (ecosystem services) or transmission back to livestock (ecosystem disservices) by vultures, thus contributing respectively to disease mitigation or amplification in natural and farmed environments. The results show a seasonal high occurrence and turnover of a high variety of serotypes, especially swine-adapted ones isolated at high frequency. This suggests that vultures can be reservoirs and long-distance carriers of faecal Salmonella shed in supplementary feeding stations and breeding colonies. Contrary to their conservation purposes, feeding stations can act as Salmonella hotspots and reservoirs. However, a role for vultures in the transmission back to food-producing animals seems impossible because they do not come into contact at indoor farms, while transmission to free-ranging ruminants was not supported by the presence of ruminant-adapted serotypes in the vultures. Therefore, vultures do not promote disservices associated with the re-infection of livestock with Salmonella, but can provide quantitatively important ecosystem services by removing carrion contaminated with these and other zoonotic pathogens potentially affecting their health. Sanitary vigilance of the farms authorised to provide food for avian scavengers should avoid the disposal of swine and poultry carcasses with Salmonella and the antibiotics used to treat it. Extensive free-ranging livestock and their carcasses exploited in the countryside should be a priority for the conservation of vultures and their ecological function as cleaners and disease mitigators.

Prevalence, Molecular Characterization and Antimicrobial Susceptibility of Clostridioides difficile Isolated from Pig Carcasses and Pork Products in Central Italy.

In the last decade, the incidence and severity of Clostridioides difficile infections (CDIs) in humans have been increasing and community-associated infections have been described. For these reasons, the interest in C. difficile in food and in food animals has increased, suggesting other possible sources of C. difficile acquisition. This study evaluated the presence of C. difficile on pig carcasses at the slaughterhouse and in pork products in Central Italy. The contamination rate on pig carcasses was 4/179 (2.3%). Regarding food samples, a total of 216 pork products were tested (74 raw meat preparations and 142 ready-to-eat food samples made by cured raw meat). The real-time PCR screening was positive for 1/74 raw meat preparation (1.35%) and for 1/142 ready-to-eat food samples (0.7%) C. difficile was isolated only from the raw meat preparation (pork sausage). All the isolated strains were toxigenic and susceptible to all the tested antibiotics. Strains isolated from carcass samples displayed A+B+CDTa+CDTb+ profile, were toxinotype IV and belonged to the same ribotype arbitrary named TV93, while the one isolated from food samples displayed A+B+CDTa-CDTb- profile and it was not possible to determine ribotype and toxinotype, because it was lost after freeze storage. It was concluded that the prevalence of C. difficile in the pork supply chain is very low.

Authentication of the Montanera Period on Carcasses of Iberian Pigs by Using Analytical Techniques and Chemometric Analyses.

The potential of two complementary analytical techniques (near infrared spectroscopy, NIRS and gas chromatography-ion mobility spectrometry, GC-IMS) was used to establish the time that Iberian pigs have been fed on acorns and pasture and to verify their genetic purity. For both techniques it was neither necessary to carry out any chemical treatment in advance nor to identify individual compounds. The results showed that both the NIR spectrum and the spectral fingerprint obtained by GC-IMS were affected by the time that the Iberian pig feeds on natural resources. High percentages of correct classification were achieved in the calibration for both techniques: >98% for the days of montanera and >96% for the breed by NIRS and >99% for the days of montanera and >98% for the breed by GC-IMS. The results obtained showed that NIR spectra taken from intact samples is a quick classification method according to the time of montanera and breed.

A comparative study of the entomofauna (Coleoptera, Diptera) associated with hanging and ground pig carcasses in a forest habitat of Poland.

Suicide by hanging is one of the most common causes of death, worldwide. Therefore, studies of insects associated with hanging cadavers are important for developing entomological tools capable of estimating the postmortem intervals in such cases. We conducted a two-year experiment, in the spring, summer, and autumn of 2012 and 2013, studying the entomofauna of 12 hanging and 12 ground, decomposing pig carcasses in a hornbeam-oak forest, in western Poland. Both carcass treatments showed colonization by similar insect species. Among the 224 taxa collected, 61 and 52 were minimally abundant (≥10 individuals) during the adult stage on at least one hanging and at least one ground carcass, respectively. During the spring and autumn, more adult flies were attracted to hanging carcasses, whereas during the summer, their numbers were similar on both hanging and ground carcasses. Adult beetles were higher in number on hanging carcasses during the spring, but during the summer and autumn, they were slightly more abundant on ground pig carcasses. Irrespective of the carcass treatment, Dryomyzidae, Muscidae, and Piophilidae were the most abundant flies, whereas representatives of Geotrupidae, Histeridae, and Silphidae were the dominant families in the studied beetle community. Only Anoplotrupes stercorosus (Scriba) and Hydrotaea similis Meade adults and Calliphora vomitoria (Linnaeus) larvae reached minimal abundance on all carcasses during all seasons. Carcass treatment only affected the numbers of minimally abundant fly species at the larval stage due to their higher numbers on ground pig carcasses. Although differences in the residency patterns of insects were observed between carcass treatments, they were not significant. In general, adult insects tended to be present for slightly longer times on hanging carcasses than on ground carcasses, whereas beetle larvae had longer presences on ground pig carcasses. Fly larvae were present on ground carcasses for the same amount of time as on hanging carcasses. More broken residency patterns were observed among the adult insects and beetle larvae that colonized hanging carcasses. Fly larvae that colonized ground carcasses had more breaks within the presence periods than fly larvae colonizing hanging carcasses. Neither treatment significantly affected the time of insect appearance on carrion. Most species appeared at the same time or at within one- or two-day intervals on both treatments. Seasonal differences in insect appearance were found, with most species colonizing quickly during the summer. The implications of these findings for forensic entomology are discussed.

Decomposition of hanging pig carcasses in a forest habitat of Poland.

We present the results of the first study on the decomposition of hanging carcasses in a forest habitat of Poland. The decomposition of 12 hanging pigs and 12 ground pigs, which were used as control, in spring, summer, and autumn was examined during a two-year study. Mosaic decomposition was observed with bloating, active decay, and advanced decay irrespective of the carrion treatment (hanging vs ground). Bloating was less visible or did not occur at all in hanging pigs. After undergoing advanced decay, carcasses became mummified and remained in the hanging position. Complete skeletonization was observed only within the head in one case in summer. Irrespective of the carrion treatment, the active decay was driven by the same blow fly species with a similar seasonal pattern of larval dominance. The season was found to affect the decomposition of the hanging carcasses as revealed by a distinct delay in the onset of bloating in autumn and the late onset of active decay in spring. The duration of active decay was the shortest in summer and the longest in autumn. The effect of carrion treatment on the rate of decomposition was statistically insignificant. Nevertheless, some differences in the duration of decomposition between hanging and ground pigs were observed. The most distinct difference was that earlier decomposition proceeded faster in hanging pigs in spring and a delayed decomposition was observed in autumn. In summer, decomposition was similar in all the carcasses. The presented results are largely in contrast to the findings of previous studies on decomposition of hanging pig carrion.

Supplementary feeding as a source of multiresistant Salmonella in endangered Egyptian vultures.

Wild birds have repeatedly been highlighted as vectors in the dissemination of livestock and human pathogens. Here, the occurrence, serotypes and antimicrobial resistance of Salmonella were assessed in adult Egyptian vultures (Neophron percnopterus), to test the hypothesis that infection is associated with the consumption of swine carcasses provided at supplementary feeding stations (SFSs). Faeces of year-round resident griffon vultures (Gyps fulvus) were also tested to assess whether infection was acquired in the breeding grounds of both species or in the African wintering quarters of Egyptian vultures. Depending on the shedding rate criteria considered, the occurrence of infection in Egyptian vultures varied between the three consecutive sampling days in a range with a minimum of 23%-41% and a maximum of 64%-92% of individuals (n = 11-14 individuals, 27-39 faeces). The occurrence in the single sampling of griffon vultures was 61% of faeces (n = 18). Vultures mostly fed on pig carcasses, which together with their predominant infection with multiresistant serotypes (mostly the monophasic 4,12:i:- variant resistant to aminopenicillins, aminoglycosides and tetracyclines) typically found in pigs from Spain, strongly supports a carcass-to-vulture transmission and cross-infection routes at SFSs. Efforts are encouraged to avoid discarding carcasses of pigs with Salmonella at SFSs established for the conservation of threatened scavengers. This could contribute to reducing the long-distance transmission of resistant pathogens with an impact on livestock and human health while avoiding infection risk and its effects on wildlife.

Characterization of odor emissions and microbial community structure during degradation of pig carcasses using the soil burial-composting method.

A soil burial-composting method was proposed as a hybrid disposal method for infected carcasses. This is a modified soil burial technique that involves covering carcasses with compost to achieve a final compost bed of 1.0-1.2 m during the soil burial process. To evaluate the feasibility and applicability of the soil burial-composting method, a pilot-scale system was constructed to dispose of pig carcasses and monitored its performance for 346 days. Temperature around the pig carcasses in the compost bed increased gradually, and was in the range of 35-45 °C after 200 days. Mesophilic (Sporosarcina and Steroidobacter) and thermophilic (Truepera) bacteria were dominant in the compost bed. Based on odor gas profiling and the morphological properties of the carcasses excavated after 346 days, it was estimated that an advanced decay stage was reached after 243 days. Considering the results of previous studies, the carcass degradation rate achieved by soil burial-composting was faster than that of soil burial, but slower than that of the composting method. Sum of odor quotient (SOQ) in the upper soil bed was lower than the SOQ in the compost bed where the carcasses were buried. This result demonstrated that the upper soil bed functioned as a biofilter to mitigate odor gases emitted during degradation of the carcasses. The soil burial-composting disposal method is preferred over soil burial because the degradation of carcasses is faster, and over composting because odor complaints and compost usage can be minimized.

Culture- and molecular-based detection of swine-adapted Salmonella shed by avian scavengers.

Salmonella can play an important role as a disease agent in wildlife, which can then act as carriers and reservoirs of sanitary importance at the livestock-human interface. Transmission from livestock to avian scavengers can occur when these species consume contaminated carcasses and meat remains in supplementary feeding stations and rubbish dumps. We compared the performance of PCR-based detection with conventional culture-based methods to detect Salmonella in the faeces of red kites (Milvus milvus) and griffon vultures (Gyps fulvus) in central Spain. The occurrence of culturable Salmonella was intermediate in red kites (1.9%, n=52) and high in griffon vultures (26.3%, n=99). These proportions were clearly higher with PCR-based detection (13.5% and 40.4%, respectively). Confirmation cultures failed to grow Salmonella in all faecal samples positive by the molecular assay but negative by the initial conventional culture in both scavenger species, indicating the occurrence of false (non-culturable) positives by PCR-based detection. This suggests that the molecular assay is highly sensitive to detecting viable Salmonella in cultures, but also partial genomes and dead or unviable bacteria from past infections or contamination. Thus, the actual occurrence of Salmonella in a particular sampling time period can be underestimated when using only culture detection. The serovars found in the scavenger faeces were among the most frequently isolated in pigs from Spain and other EU countries, especially those generally recognized as swine-adapted monophasic variants of S. Typhimurium. Because the studied species obtain much of their food from pig carcasses, this livestock may be the primary source of Salmonella via direct ingestion of infected carcasses and indirectly via contamination due to the unsanitary conditions found in supplementary feeding stations established for scavenger conservation. Combining culture- and molecular-based detection is encouraged to understand the epidemiology and impact of Salmonella in wildlife populations.

Toxoplasma gondii Lineages Circulating in Slaughtered Industrial Pigs and Potential Risk for Consumers.

Toxoplasma gondii is a cosmopolitan zoonotic protozoan parasite, and the consumption of raw or undercooked pig meat is one of the most important sources of T. gondii infection. Three predominant lineages, types I, II, and III, are widespread in Europe. Although still poorly understood, a relationship between each type and the severity of illness represents a public health issue. To gain further knowledge of the genotypes in circulation and of the potential risk for consumers, one heart sample and one diaphragm sample (206 total) were taken from each of 103 pig carcasses at an abattoir in Italy. Then, we used 529-bp repetitive element PCR and a B1 real-time PCR high-resolution melting assay coupled with sequencing to detect and genotype T. gondii isolates. T. gondii DNA was detected in 14 pigs (13.6%, 95% confidence interval = 7 to 20.2%), and types I (3.9%), II (5.8%), and III (3.9%) were identified. We found that heart tissue had a significantly higher PCR positivity rate for T. gondii than did diaphragm tissue. This is Europe's largest study on genotyping of T. gondii from pigs, and it demonstrates that all three main lineages are present in carcasses of industrially reared pigs in Italy. There is a potential risk to consumers of infection with any or all of the three lineages, and the related clinical consequences should be taken into account. This study suggests that monitoring of T. gondii types in meat is essential, especially in meat that is traditionally eaten raw or that is minimally processed.

Novel ultrasound approach for measuring marbling in pork.

An ultrasound examination was done on the m. longissimus lumborum, between the 10th and the 11th thoracic vertebra, on two sides (inside and outside the thoracic cavity) of the left half-carcasses of 162 fatteners. The carcasses were classified for lean meat percentage using the SEUROP system. The R pig carcasses (47.7% lean) had the thickest backfat (30.6mm; P≤0.01) and the highest content of intramuscular fat (IMF=2.28%; P≤0.01). More artifacts-free images were collected from the inside compared to the outside of the pig carcasses (90.1% vs. 58.6%; P≤0.01). The percent of bright pixels (PBP) was the highest for the inside, for all the lean-meat-content classes (P≤0.01). The correlation between the PBP and the IMF was higher for the images of the inside compared to the outside (r=0.811; P=0.001 vs. r=0.523; P=0.009). The ultrasound images of the inside of the carcasses proved to be the most useful for making an assessment of the marbling.

Nitrous oxide, methane and carbon dioxide dynamics from experimental pig graves.

Twelve pig carcasses were buried in single, shallow and deep (30 and 90 cm, respectively) graves at an experimental site near Ottawa, Ontario, Canada, with three shallow and three deep wrapped in black plastic garbage bags. An additional six carcasses were left at the surface to decompose, three of which were bagged. Six reference pits without remains were also dug. The objective of this three-year study was to examine the biogeochemistry and utility of nitrous oxide (N2O), methane (CH4) and carbon dioxide (CO2) in grave detection and whether grave depth or cadaver condition (bagged versus bare) affected soil pore air concentrations and emission of the three gases. Graves showed significantly higher (α=0.05) concentrations and surface fluxes of N2O and CO2 than reference pits, but there was no difference in CH4 between graves and reference pits. While CH4 decreased with depth in the soil profiles, N2O and CO2 showed a large increase compared to reference pits. Shallow graves showed significantly higher emissions and pore air concentrations of N2O and CO2 than deep graves, as did bare versus bagged carcasses.

Decomposition Rate and Pattern in Hanging Pigs.

Accurate prediction of the postmortem interval requires an understanding of the decomposition process and the factors acting upon it. A controlled experiment, over 60 days at an outdoor site in the northwest of England, used 20 freshly killed pigs (Sus scrofa) as human analogues to study decomposition rate and pattern. Ten pigs were hung off the ground and ten placed on the surface. Observed differences in the decomposition pattern required a new decomposition scoring scale to be produced for the hanging pigs to enable comparisons with the surface pigs. The difference in the rate of decomposition between hanging and surface pigs was statistically significant (p=0.001). Hanging pigs reached advanced decomposition stages sooner, but lagged behind during the early stages. This delay is believed to result from lower variety and quantity of insects, due to restricted beetle access to the aerial carcass, and/or writhing maggots falling from the carcass.

Investigation of Listeria monocytogenes, Salmonella enterica and Yersinia enterocolitica in pig carcasses in Southern Brazil / Pesquisa de Listeria monocytogenes, Salmonella enterica and Yersinia enterocolitica em carcaças de suínos no sul do Brasil

The intensification of pig production and advances in the sanitary control of herds profoundly changed the profile of risk attributed to pork consumption. In the actual scenario, most microorganisms related to macroscopic lesions observed in the post mortem inspection are not transmitted by food, while foodborne bacteria of importance to consumer health do not cause macroscopic lesions. In Brazil, the "Ministério da Agricultura, Pecuária e Abastecimento" requested a scientific opinion on the prioritizing of pathogens potentially transmitted by unprocessed pork. After conducting a qualitative risk assessment, only Salmonella enterica was classified as of high risk to consumers. The present study was part of the validation step of the risk assessment and aimed to investigate the frequency of S. enterica, Yersinia enterocolitica and Listeria monocytogenes and hygienic-sanitary indicators in pig carcasses of pigs rose under intensive production and slaughtered under the Federal Inspection System in three slaughterhouses located in Southern Brazil. Additionally, the antimicrobial resistance profile of the isolated pathogens was also investigated. A total of 378 carcasses were sampled by superficial sponges before the chilling step in three slaughterhouses. Samples were investigated for the presence of the three aforementioned pathogens and subjected to enumeration of Colony Formation Units (log CFU.cm-1) of total aerobic mesophiles (TAM) and Enterobacteriaceae. Salmonella strains were tested by disc diffusion test for resistance to eleven antimicrobials. There were significantly statistical differences (p<0.0001) on the median counts of both indicators between the slaughterhouses. The median of TAM was very close for Slaughterhouses A and B: 1.573 log CFU.cm-1 and 1.6014 log CFU.cm-1, respectively. While in Slaughterhouse C, a higher TAM median was detected (2.216 log CFU.cm-1). A similar profile was observed regarding to Enterobacteriaceae, and medians were calculated as follow: -0.426 log CFU.cm-1 in Slaughterhouse A; 0.2163 log CFU.cm-1 in B; and 0.633 log CFU.cm-1 in C. Regarding the pathogens investigated, L. monocytogenes was not detected and only one carcass from Slaughterhouse C was positive for Y. enterocolitica. Thus, the results suggest a very low prevalence of L. monocytogenes and Y. enterocolitica in the sampled population. A total of 65 (17.2%) carcasses were positive for S. enterica, with a difference in frequencies between slaughterhouses and slaughter days. The prevalence of Salmonella positive carcasses was higher in the Slaughterhouse C (25.4%; CI 95% 19-32%) in comparison with A (9.5%; CI 95% 9-14%) and B (18.3%; CI 95% 12-24%). There was no significantly statistical association between Enterobacteriaceae counts and Salmonella isolation on carcass surface (p=0.69). The slaughtering day, nested within the slaughterhouse, explains 31.3% of Salmonella prevalence variability. S. Typhimurium (38.1%) was the most prevalent, followed by S. Infantis (30.1%). Among the 61 Salmonella strains tested for resistance to antimicrobials, 18 (31.6%) were full-susceptible. No strain displayed resistance to azithromycin, ceftazidime, cefotaxime and meropenem. The highest resistance frequency was displayed to tetracycline (54.1%), followed by ampicillin (50.82%), nalidixic acid (42.62%) and chloramphenicol (42.62). Multi-resistance was detected in 52.54% of the, strains. In conclusion, S. enterica is more prevalent in pre-chill pig carcasses than Y. enterocolitica and L. monocytogenes and thus should be prioritized in monitoring and control programs at slaughter. Salmonella serovars varied among slaughterhouses and present significant differences in their resistance to antimicrobials. Slaughterhouses that present higher medians of TAM or Enterobacteriaceae in a monitoring period may have higher S. enterica prevalences as well. However, there is a high variation of S. enterica prevalence among slaughter days, which cannot be always related to the hygienic indicators counts observed on a given day.(AU)

A intensificação da produção de suínos e os avanços no controle sanitário dos rebanhos alterou de forma importante o perfil de risco do consumo de carne suína. No cenário atual, a maioria dos microrganismos causadores de lesões macroscópicas detectáveis na inspeção post mortem não são transmissíveis por alimentos, enquanto bactérias de importância como causadoras de doenças transmitidas por alimentos não causam lesões macroscópicas. No Brasil, o Ministério da Agricultura, Pecuária e Abastecimento solicitou uma opinião científica sobre a priorização de patógenos potencialmente transmitidos pela carne suína in natura. Após conduzir uma avaliação de risco qualitativa, apenas Salmonella enterica foi classificada como de alto risco para o consumidor. O presente estudo foi parte da etapa de validação da avaliação de risco e objetivou: investigar a frequência de S. enterica, Yersinia enterocolitica e Listeria. monocytogenes; e enumerar indicadores higiênico-sanitários em carcaças de suínos abatidos sob inspeção federal em frigoríficos dedicados ao abate de suínos sob sistema intensivo de criação no sul do Brasil. Além disso, o perfil de resistência a antimicrobianos dos patógenos isolados foi investigado. A superfície de um total de 378 carcaças foi amostrada por esponjas, na etapa de pré-resfriamento em três matadouros frigoríficos (A, B, C). As amostras foram investigadas quanto à presença dos três patógenos acima mencionados e quanto à enumeração de Unidades Formadoras de Colônia (log UFC.cm-1) de mesófilos aeróbios totais (MAT) e Enterobacteriaceae. As cepas isoladas de Salmonella foram testadas quanto à resistência a onze antimicrobianos pela técnica de disco difusão. As medianas de contagem de ambos os indicadores apresentaram diferença significativa (p<0,0001) entre matadouros-frigoríficos. A mediana de MAT foi bastante próxima para A e B (1,573 log UFC.cm-1 e 1,6014 log UFC.cm-1, respectivamente), enquanto em C uma mediana de MAT mais elevada foi determinada (2,216 log CFU.cm-1). Um perfil semelhante foi observado em relação a Enterobacteriaceae, sendo as medianas calculadas para A, B e C, respectivamente: -0,426 log CFU.cm-1; 0,2163 log UFC.cm-1; e 0,633 log UFC.cm-1. Em relação aos patógenos investigados, L. monocytogenes não foi detectada e apenas uma carcaça, do Matadouro C, foi positiva para Y. enterocolitica. Portanto, os resultados sugerem uma prevalência muito baixa desses patógenos na população amostrada. Em um total de 65 (17,2%) carcaças houve isolamento de S. enterica, com diferença nas frequências observadas entre matadouros e dias de abate. A prevalência de carcaças positivas para S. enterica foi maior no Matadouro C (25,4%; IC95% 19-32%) em comparação com A (9,5%; IC95% 9-14%) e B (18,3%; IC95% 12-24%). Não houve associação estatística entre o número de Enterobacteriaceae e o isolamento de S. enterica na superfície das carcaças (p=0,69). O dia de abate agrupado por frigorífico explica 31,3% da variação na prevalência de Salmonella. O sorovar mais frequente de S. enterica foi Typhimurium (38,1%) seguido de S. Infantis (30,1%). Entre as 61 cepas de S. enterica testadas quanto à resistência a antimicrobianos, 18 (31,6%) foram totalmente suscetíveis aos antimicrobianos testados. Nenhuma cepa apresentou resistência a azitromicina, ceftazidima, cefotaxima e meropenem. As maiores frequências de resistência foram demonstradas contra tetraciclina (54,1%), ampicilina (50,8%), ácido nalidíxico (42,62%) e cloranfenicol (42,62%). Em 52,54% das cepas foi detectada multi-resistência. Em conclusão, S. enterica é mais prevalente em carcaças suínas no pré-resfriamento do que Y. enterocolitica e L. monocytogenes. Portanto, S. enterica deve ser priorizada em programas de monitoramento e controle ao abate. Os sorovares de Salmonella variam entre matadouros e apresentam diferenças significativas na resistência a antimicrobianos. Matadouros de suínos que apresentam medianas de MAT e Enterobacteriaceae num período de monitoramento podem apresentar também prevalências mais de altas de presença de S. enterica. Entretanto, há uma alta variabilidade na frequência de S. enterica entre dias de abate, e nem sempre há relação entre essa frequência e a contagem de indicadores higiênico-sanitários determinados num determinado dia.(AU)