Investigation of the presence and seasonal prevalence of Salmonella spp., Salmonella Typhimurium, and its monophasic variant I 4,5,12:i:- in United States swine feed mills

Master of ScienceFood Science InstituteValentina TrinettaSalmonella is an important pathogen of public health concern. Each year, Salmonella costs the food industry approximately $2.3 billion. In recent years, the number of cases of Salmonella linked to pork products has also increased in the United States (US). Although pork has the lowest association with human foodborne illness when compared to beef and chicken, it is the most consumed meat in the world. Therefore, Salmonella is a significant food safety concern for the American swine industry. This pathogen can be present along all the food production chain from farm to fork and recent studies reported the isolation of Salmonella enterica serovar Typhimurium (ST) and its monophasic variant 4,[5],12:i:- (STM) in feed and feed ingredients. The occurrence of these pathogens in the pre-harvest environment can translate to entry and contamination of the human food chain. Nevertheless, little is known about Salmonella incidence and association with these types of environments. Hence, the purpose of this study was to investigate the presence and seasonal prevalence of Salmonella spp., ST and STM in selected feed mills, among the major US swine feed production areas. Eleven swine feed mills in eight different states were selected. Six mills produced only mash feed, while the other five facilities produced both mash and pelleted feed. Visits were conducted during fall 2016, early spring 2017 and summer 2017. Twelve environmental samples were collected within each facility and season, representative of the production flow, from receiving of ingredients to the finished product, including floor surfaces, equipment dust, workers’ shoes, and finished feed. Samples were analyzed following the USDA-FSIS guidelines and culture positive samples were analyzed by PCR. A multiplex PCR assay was also performed to differentiate Salmonella Typhimurium and Salmonella I 4,[5],12:i:- from the other serotypes. Associations between mill, season, mill type, sample site and Salmonella prevalence were analyzed using generalized linear mixed models (P < 0.05). From the 383 samples collected, 49 (12.8%) were identified as Salmonella spp.; two (5.1%) were identified from feed, while the other 47 (13.7%) originated from equipment or surfaces. Two samples were positive for ST and three for STM by multiplex PCR. Mill (P = 0.003) and season (P = 0.006) were statistically associated with the presence of Salmonella, with higher prevalence in fall and summer (13.2%) as compared to spring (3.6%). These findings demonstrate the seasonal prevalence of Salmonella spp., ST and STM in feed mills across the US, highlighting the potential role of the feed mill environment as a microbial entry route into the human food chain. The data presented can be also used as a tool to assist in the implementation of mitigation strategies for pre-harvest food safety

Evaluation of Salmonella presence in selected United States feed mills

Abstract Salmonella is a pathogen of public health concern. Each year, Salmonella infections cost to the food industry approximately $2.3 billion and 33% of the reported cases are associated with beef, poultry, or pork. Pathogen presence in feed mills can represent one of the many potential routes for entry and transmission into the food production chain. Nevertheless, little is known about Salmonella incidence and association with these types of environments. The objective of this study was to investigate Salmonella presence in different feed mills across the United States. Eleven facilities were selected in eight states and 12 sites were sampled within each feed mill. Samples were analyzed following the FSIS guidelines for isolation and identification of Salmonella. Positive isolates were further investigated by a PCR analysis targeting the invA gene to differentiate for Salmonella enterica. The total number of environmental samples collected was 237: 66% resulted culture positive and 13.1% were PCR positive. All sampled feed mills had at least one culture positive site and following production flow the number of positive samples decreased from ingredient receiving to final product. These preliminary results demonstrate the presence of Salmonella in selected United States feed mills and suggest their potential role as vehicle for pathogen transmission and spread into the food production chain

Comparison data of a two-target real-time PCR assay with and without an internal control in detecting Salmonella enterica from cattle lymph nodes

A real-time PCR (qPCR) assay targeting on invA and pagC genes was developed and validated for the detection and quantification of Salmonella enterica strains (Bai et al., 2018) [1]. A host gene, normally an endogenous housekeeping gene (Beer-Davidson et al., 2018; Poon et al., 2004) [2,3], or an irrelevant exogenous gene (Cheng et al., 2015; Sedlak et al., 2014) [4,5] has been widely used as an internal control to monitor nucleic acid extraction efficiencies and potential PCR inhibitions in PCR-based detection assays. An endogenous internal control designed based on the 18S rRNA gene was used in the above-mentioned qPCR assay. This 18S rRNA internal control amplifies the target gene in multiple species including bovine, swine, ovine, caprine and cervine. Data was generated by the duplex qPCR assay on 138 enriched cattle lymph node samples without the internal control, and compared with data on the same samples tested by the triplex qPCR assay that has the 18S rRNA gene as internal control. Threshold cycle (Ct) data for the duplex and the triplex qPCR on the 138 samples were similar, and are presented in this brief report. Keywords: Real-time PCR, Threshold cycle, Internal control, Salmonella, Foodborne pathogen, Lymph nod