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.

Prevalence and Persistence of Multidrug-Resistant Yersinia enterocolitica 4/O:3 in Tonsils of Slaughter Pigs from Different Housing Systems in Croatia.

Yersinia enterocolitica is one of the priority biological hazards in pork inspection. Persistence of the pathogen, including strains resistant to antimicrobials, should be evaluated in pigs from different housing systems for risk ranking of farms. In this 2019 study, tonsils were collected from 234 pigs, of which 69 (29.5%) were fattened on 3 big integrated farms, 130 (55.5%) on 10 medium-sized farms, and 35 (15%) on 13 small family farms. In addition, 92 pork cuts and minced meat samples from the same farms were tested for the presence of Y. enterocolitica using the culture method. Phenotypic and genetic characteristics of the isolates were compared with previously collected isolates from 2014. The overall prevalence of Y. enterocolitica in pig tonsils was 43% [95% CI 36.7−49.7]. In pigs from big integrated, medium-sized, and small family farms, the prevalence was 29%, 52%, and 40%, respectively. All retail samples of portioned and minced pork tested negative for pathogenic Y. enterocolitica, likely due to high hygienic standards in slaughterhouses/cutting meat or low sensitivity of culture methods in these matrices. The highest recovery rate of the pathogen from tonsils was found when alkali-treated PSB and CIN agar were combined. The biosecurity category of integrated and medium farms did not affect the differences in prevalence of Y. enterocolitica (p > 0.05), in contrast to family farms. Pathogenic ail-positive Y. enterocolitica biotype 4 serotype O:3 persisted in the tonsils of pigs regardless of the type of farm, slaughterhouse, and year of isolation 2014 and 2019. PFGE typing revealed the high genetic concordance (80.6 to 100%) of all the Y. enterocolitica 4/O:3 isolates. A statistically significant higher prevalence of multidrug-resistant Y. enterocolitica 4/O:3 isolates was detected in the tonsils of pigs from big integrated farms compared to the other farm types (p < 0.05), with predominant and increasing resistance to nalidixic acid, chloramphenicol, and streptomycin. This study demonstrated multidrug resistance of the pathogen in pigs likely due to more antimicrobial pressure on big farms, with intriguing resistance to some clinically relevant antimicrobials used in the treatment of yersiniosis in humans.

Mycobiota and Mycotoxin Contamination of Traditional and Industrial Dry-Fermented Sausage Kulen.

The aim of this study was to identify and compare surface mycobiota of traditional and industrial Croatian dry-fermented sausage Kulen, especially toxicogenic species, and to detect contamination with mycotoxins recognized as the most important for meat products. Identification of mould species was performed by sequence analysis of beta- tubulin and calmodulin gene, while the determination of mycotoxins aflatoxin B1 (AFB1), ochratoxin A (OTA), and cyclopiazonic acid (CPA) was carried out using the LC-MS/MS (liquid chromatography-tandem mass spectrometry) method. The results showed a significantly higher number of mould isolates and greater species (including of those mycotoxigenic) diversity in traditional Kulen samples in comparison with the industrial ones. P. commune, as a potential CPA-producer, was the most represented in traditional Kulen (19.0%), followed by P. solitum (16.6%), which was the most represented in industrial Kulen samples (23.8%). The results also showed that 69% of the traditional sausage samples were contaminated with either CPA or OTA in concentrations of up to 13.35 µg/kg and 6.95 µg/kg, respectively, while in the industrial samples only OTA was detected (in a single sample in the concentration of 0.42 µg/kg). Mycotoxin AFB1 and its producers were not detected in any of the analysed samples (

Tracing of Listeria monocytogenes Contamination Routes in Fermented Sausage Production Chain by Pulsed-Field Gel Electrophoresis Typing.

In this study, the presence of Listeria monocytogenes was assessed along the production process of fermented sausages in a small-scale facility. Following the isolation of the pathogen from the final product (ISO 11290-1), retrospective sampling was performed during the production of a new batch of sausages, including raw materials, casings, additives, sausage mixtures, sausages during fermentation, and environmental samples. L. monocytogenes was recovered from the following sampling points: the defrosting room and the cuttering, mixing, stuffing, and fermentation phases. Ten strains were isolated, molecularly confirmed as L. monocytogenes by means of a molecular detection system, and subjected to pulsed-field gel electrophoresis (PFGE) typing. On the basis of an unweighted pair group method with arithmetic mean (UPGMA) dendrogram from Ascl pulsotypes, the strains were indistinguishable (no band difference). The same pulsotypes of strains present in both batches of sausages, as well as in environmental samples, indicated the persistence of L. monocytogenes in the sausage production unit.

Effect of Fungal Solid-State Fermented Product in Broiler Chicken Nutrition on Quality and Safety of Produced Breast Meat.

The aim of this work was to analyse the effect of addition of 10% (w/w) fermented bioproduct into commercial broiler feed on fatty acid profile, lipid oxidative stability, and sensory properties of chicken breast meat. The fermented bioproduct was prepared by fermentation of cornmeal by filamentous fungi Umbelopsis isabellina CCF 2412 in solid-state fermentation (SSF) process and the final bioproduct was enriched with gamma-linolenic acid and beta-carotene. In the experiment, 80 pieces of 1-day-old chickens COBB 500 were used. Half of them (control group) were fed only with commercial feed. Chickens of the experimental group were fed with commercial feed, and, from the 11th day of age until the time of slaughter (39th day), 10% of commercial feed was replaced with fermented bioproduct. Application of fermented bioproduct into commercial feed mixture positively influenced profile of fatty acids in breast meat. The amount of gamma-linolenic, alpha-linolenic, and oleic acids in fat of breast muscles was increased and n-6/n-3 ratio was significantly decreased. Profile and content of PUFAs did not change after thermal treatment of meat. Oxidative stability of fat and sensory properties of the meat during the storage (4°C, 7 days) of meat were not affected by fermented bioproduct.

Lactobacillus plantarum with Functional Properties: An Approach to Increase Safety and Shelf-Life of Fermented Foods.

Lactobacillus plantarum (widespread member of the genus Lactobacillus) is one of the most studied species extensively used in food industry as probiotic microorganism and/or microbial starter. The exploitation of Lb. plantarum strains with their long history in food fermentation forms an emerging field and design of added-value foods. Lb. plantarum strains were also used to produce new functional (traditional/novel) foods and beverages with improved nutritional and technological features. Lb. plantarum strains were identified from many traditional foods and characterized for their systematics and molecular taxonomy, enzyme systems (α-amylase, esterase, lipase, α-glucosidase, ß-glucosidase, enolase, phosphoketolase, lactase dehydrogenase, etc.), and bioactive compounds (bacteriocin, dipeptides, and other preservative compounds). This review emphasizes that the Lb. plantarum strains with their probiotic properties can have great effects against harmful microflora (foodborne pathogens) to increase safety and shelf-life of fermented foods.

Prevalence of Salmonella spp. and Yersinia enterocolitica in/on tonsils and mandibular lymph nodes of slaughtered pigs.

A total of 156 tonsils and 156 mandibular lymph nodes from fattening pigs originating from 13 farms were sampled in Croatian slaughterhouses and examined for Salmonella spp. (n=78 per organ) and Yersinia enterocolitica (n=78 per organ) by cultural methods. Salmonella was isolated from two tonsils only, both originated from animals from the same farm (5.12%), while Y. enterocolitica were recovered from 26 tonsils (33.33%) which could be traced back to 10 farms. Salmonella was absent in mandibular lymph nodes, and Y. enterocolitica was isolated from eight lymph nodes (10.25%) which originated from six farms. Y. enterocolitica was present inside the lymph nodes of two pigs. The high prevalence of Y. enterocolitica in/on pig tonsils could be the result of cross-contamination during splitting the carcasses with head. This procedure may result in higher prevalence of Y. enterocolitica on surface of mandibular lymph nodes than in their depth. Traditional veterinary postmortem examination of pig halves will not necessarily contribute to cross-contamination with Salmonella or Yersinia under conditions of present slaughter practice.

European validation of a real-time PCR-based method for detection of Listeria monocytogenes in soft cheese.

The classical microbiological method for detection of Listeria monocytogenes requires around 7 days for final confirmation, and due to perishable nature of RTE food products, there is a clear need for an alternative methodology for detection of this pathogen. This study presents an international (at European level) ISO 16140-based validation trial of a non-proprietary real-time PCR-based methodology that can generate final results in the following day of the analysis. This methodology is based on an ISO compatible enrichment coupled to a bacterial DNA extraction and a consolidated real-time PCR assay. Twelve laboratories from six European countries participated in this trial, and soft cheese was selected as food model since it can represent a difficult matrix for the bacterial DNA extraction and real-time PCR amplification. The limit of detection observed was down to 10 CFU per 25 of sample, showing excellent concordance and accordance values between samples and laboratories (>75%). In addition, excellent values were obtained for relative accuracy, specificity and sensitivity (82.75%, 96.70% and 97.62%, respectively) when the results obtained for the real-time PCR-based methods were compared to those of the ISO 11290-1 standard method. An interesting observation was that the L. monocytogenes detection by the real-time PCR method was less affected in the presence of Listeria innocua in the contaminated samples, proving therefore to be more reliable than the reference method. The results of this international trial demonstrate that the evaluated real-time PCR-based method represents an excellent alterative to the ISO standard since it shows a higher performance as well as reduce the extent of the analytical process, and can be easily implemented routinely by the competent authorities and food industry laboratories.

Dietary exposure assessment of streptomycin and tetracycline in food of animal origin on the Croatian market.

Residual antibacterials in food constitute a risk to human health, particularly because they can contribute to the transmission of antibiotic-resistant pathogenic bacteria through the food chain. This paper presents dietary exposure assessment of streptomycin and tetracycline, based on combining food consumption data in Croatia with data on the concentration of veterinary drugs in analysed samples of food of animal origin. According to the median values, the estimated daily intake of streptomycin and tetracycline through food is 11.9 and 0.7 µg/person/day, respectively. The largest contribution to streptomycin intake comes from meat (4.8 µg/person/day, i.e. 41%), but milk is the largest source for tetracycline (0.3 µg/person/day, i.e. 46%). The estimated dietary exposure to these veterinary drugs does not exceed relevant toxicological reference values and the level of exposure is assessed to be acceptable.

Microbial and physicochemical succession in fermented sausages produced with bacteriocinogenic culture of Lactobacillus sakei and semi-purified bacteriocin mesenterocin Y.

The influence of the bacteriocinogenic culture Lactobacillus sakei (10(5)/g) and semi-purified bacteriocin mesenterocin Y (2560AU/kg) on the safety and quality of traditional Croatian fermented sausages was investigated. The addition of Lb. sakei and/or mesenterocin Y reduced microbial counts (P<0.05) in the final products. After 28 days of ripening, coagulase-negative cocci decreased 1.5-2.0log, yeasts 1.2-1.4log and enterococci 1.7-2.7log. In the case of the addition of Lb. sakei, the lactic acid bacteria count was significantly (P<0.05) higher at day 7 of ripening, and was accompanied by a lower pH and a higher amount of lactic acid (P<0.05). In the final product the amount of acetic acid was significantly lower. More intensive proteolysis and an increase in ammonia content were found at the beginning of fermentation, and in the second phase of ripening in the control samples, respectively. The free fatty acid concentration was significantly lower during the entire ripening process compared to the control (P<0.05). Semi-purified mesenterocin Y did not affect the sensory properties of the sausages, whilst the addition of Lb. sakei enhanced them.