Evaluation of Virulence of Salmonella Infantis and Salmonella Enteritidis with In Vitro and In Vivo Models.

Salmonella Infantis and Enteritidis serovars have been reported as important causes of salmonellosis in humans worldwide. However, the virulence of these two serovars has yet to be compared. To evaluate the virulence of Salmonella Infantis (n = 23) and Salmonella Enteritidis (n = 7), we used two models: the Caco2 cells model (in vitro) and the Galleria mellonella model (in vivo). Additionally, the virulence genes of all tested strains were contrasted with phenotypic outcomes. Results showed that adhesion means were 18.2% for Salmonella Enteritidis and 38.2% for Salmonella Infantis strains. Invasion means were 77.1% for Salmonella Enteritidis and 56.2% for Salmonella Infantis strains. Significant differences were found between serovars in adherence and invasion assays. Mortality rates (58% for Salmonella Enteritidis and 62.6% for Salmonella Infantis) were not significantly different between serotypes. The distribution of virulence genes showed that genes fae (fimbrial adherence determinants) and shdA (nonfimbrial adherence determinants) were only found in Salmonella Infantis strains. On the other hand, the rck gene (invasion) and Plasmid-encoded fimbriae genes (pef A, B, C, D) were present in Salmonella Enteritidis exclusively. In conclusion, this study shows that Salmonella Enteritidis has a higher virulence potential under experimental conditions than Salmonella Infantis. However, more studies are needed to determine the risk that Salmonella Infantis could represent compared with Salmonella Enteritidis. Moreover, other in vivo models should be considered to assess the virulence of these serovars.

Prevalence, serovars, and risk factors associated with the presence of Salmonella in pork sold in public markets in Quito, Ecuador.

Background: Salmonella enterica are bacteria that include more than 2,500 serovars. Most of these serovars have been linked to human foodborne illnesses, mainly related to poultry and pigs. Thus, these animals are considered the reservoirs of many Salmonella serovars and strains related to antibiotic resistance. This study aimed to determine the prevalence, serovars, ß-lactam resistance genes, and the risk factors associated with Salmonella enterica in pork commercialized in open markets of Quito city. Methods: For this, 165 pork meat samples were taken from municipal markets in three areas in the city. These samples were microbiologically processed following the ISO 6579-2014 standardized method. The polymerase chain reaction (PCR) test was used to identify Salmonella serotyping and resistance genes. Strains not identified by PCR were typed by the Kauffman White Le Minor scheme. A multivariate analysis was performed to identify risk factors associated with the presence of the microorganism. Results: Salmonella prevalence in pork was 9.1%. Identified serovars were 4, [5], 12: i:- (53.3%), Infantis (33.3%), and Derby (13.4%). Furthermore, the ß-lactam resistance genes bla CTX-M-65 could be identified in three S. infantis isolates. Multivariate analysis showed that temperature (above 8°C) and cutting surfaces (wood) presented significant association values. Conclusions: In conclusion, pork in traditional markets of Quito is contaminated with Salmonella enterica, whose main serovars pose a public health concern, and shows beta-lactam resistance.

Association between animal welfare indicators and microbiological quality of beef carcasses, including Salmonella spp., from a slaughterhouse in Ecuador.

BACKGROUND AND AIM: Pre-slaughter management and slaughter operations are considered critical factors for animal welfare and meat quality. Previous studies have found poor animal welfare management at municipal slaughterhouses in Ecuador, and little is known about how this affects the microbiological quality of the meat. Therefore, the aim of the study was to analyze the association of the microbiological quality of beef carcasses and animal welfare indicators in a municipal slaughterhouse in Ecuador. MATERIALS AND METHODS: Data for 6 months were collected from a municipal slaughterhouse in Ecuador. Five trained researchers were strategically located along the slaughter process. A total of 351 animals were observed with regard to welfare indicators, and their carcasses were sampled to evaluate microbiological quality. Antemortem (slipping, falling, and vocalization) and postmortem animal welfare indicators (bleed interval, pH, temperature, and bruises) were measured. To determine the total aerobic bacteria (TAB) and Escherichia coli counts and the presence of Salmonella spp., we collected samples by swabbing four different points of each carcass. The association between microbiological quality and animal welfare indicators was studied using univariate and multivariate logistic regressions. RESULTS: The mean TAB count was 5.3 log CFU/cm2, and the mean total count of E. coli was 2.4 log CFU/cm2. Salmonella spp. were isolated in 3.1% of the carcasses. An electric goad was used in all animals, 19.1% slipped at least once, and 19.9% vocalized. The mean pH of the carcasses was 7.2, and 79.2% of carcasses had bruises. Multivariate analysis showed that Salmonella spp. and the TAB count were associated with pH and the number of bruises (p = 0.01 in both cases). CONCLUSION: Although there was non-significant association between the majority of animal welfare indicators and microbiological quality, the poor management affecting animal welfare and carcass hygiene are worrisome.

Longitudinal study describing time to Salmonella seroconversion in piglets on three farrow-to-finish farms.

BACKGROUND: Pigs are frequently colonised with Salmonella enterica, and this constitutes a major risk for human salmonellosis. The infection can be assessed by the serological response of pigs to S enterica. A longitudinal study was undertaken on-farm to correctly describe this serological response and investigate factors associated with age at Salmonella seroconversion. METHODS: Three pig farms and in each farm three successive batches were considered. Per batch, 40 piglets were selected at random from 10 sows (four piglets per sow). Blood was sampled from sows one week after farrowing and from piglets at weeks 1, 6, 10, 14, 18 and 22 and at the slaughterhouse. Salmonella antibodies were detected in serum using a commercial ELISA test. Factors related to farm characteristics, batch management system, porcine reproductive and respiratory syndrome infection, and sows' Salmonella serological status were recorded to assess their effect on age at seroconversion. RESULTS: At week 1 after farrowing, 96.5 per cent of the sows had antibodies against Salmonella. The serological results of piglets at weeks 1 and 6 only were positively correlated with those of the sows. The average age at Salmonella seroconversion was 137±2.2 days (confidence interval at 95 per cent). The first seroconversions occurred from weeks 10 to 14, but most of the pigs (54.6 per cent) were seropositive at the end of the fattening period, with variations between farms and batches (28.9-75.7 per cent). Herd/farm was significantly associated with age at seroconversion. CONCLUSION: This longitudinal study allowed the authors to follow precisely the evolution of Salmonella seroconversion from maternity to slaughterhouse and confirm the relationship between the seroconversion of sows and serology of their piglets. Moreover, factors related to farm practices and management as a whole are more influential than individual factors (at the pig level) on age at Salmonella seroconversion.

Experimental infection of pigs by Salmonella Derby, S. Typhimurium and monophasic variant of S. Typhimurium: Comparison of colonization and serology.

Salmonella serovars Derby, Typhimurium and the monophasic variant of Salmonella Typhimurium are the most frequently isolated serovars in pigs in France. To compare the excretion patterns, seroconversion to Salmonella and contamination of the organs of pigs inoculated with strains of all three serovars, we conducted an experimental trial with 28 SPF piglets. Four were used as a negative control, while the other 24 were divided equally into three groups. Each group was inoculated at 7 weeks of age with a different strain: S. Derby (SDb), S. Typhimurium (ST), and the monophasic variant of S. Typhimurium (mST). Fecal and blood samples were collected twice a week up until necropsy, on 21 days post-inoculation (DPI) for half of each group and 49 DPI for the remaining piglets. During necropsy, the tonsils, mesenteric lymph nodes and various intestinal contents were collected from each pig. Salmonella bacteria were quantified in CFU/g by a bacteriological method, and levels of Salmonella antibodies were measured using an ELISA Kit. Piglets inoculated with mST continuously excreted Salmonella in their feces throughout the trial. For each of the other serovars, one piglet was Salmonella-negative on one DPI. The quantity of Salmonella excreted was statistically different between the group inoculated with ST and mST (p < 0.05), but no differences were found between the other serovars. The tonsils, cecum and jejunum were the most contaminated organs in all groups. Seroconversion for all the piglets was completed by different DPI: 28 for ST, 31 for mST and 38 for SDb. No major differences were found in terms of excretion and colonization among the studied serovars.

Colonization of Pigs Experimentally Infected with a Monophasic Variant of Salmonella Typhimurium.

The monophasic variant of Salmonella Typhimurium is highly prevalent in human and in pork. However, little is known about colonization dynamics and serology in pigs. We orally inoculated 24 seven-week-old piglets with 109 CFU/pig of a porcine strain of monophasic Salmonella Typhimurium in an experimental trial. Three groups of eight piglets were orally inoculated and monitored for 21, 49, or 84 days post-inoculation until necropsied. From 3 days post-inoculation to necropsy, individual feces were sampled twice weekly and blood once weekly. At necropsy, the tonsils, mesenteric lymph nodes, and the contents of the duodenum, jejunum, ileum, and cecum were collected from each pig. We determined the number of CFU/g in all the samples and measured also Salmonella antibodies in OD% in all blood samples. At different times during the trial, we tested by MLVA (Multilocus Variable Number Tandem Repeat Analysis) the genomic stability of the strain after passing through the intestinal tract. Salmonella was continuously excreted by pigs, ranging from 1.4 to 5.8 log10 CFU/g. At necropsy, Salmonella was present in all samples, but the tonsils were particularly infected. Salmonella antibodies were detected in five pigs 7 days post-inoculation. At 49 days post-inoculation, all the pigs were seropositive. We observed new MLVA types for 3.3% of the isolates tested over the trial. Our study allowed us to show the serovar's ability to persist in pigs after infection up to 84 days post-inoculation. We demonstrated that Salmonella seroconversion appeared earlier than in naturally infected pigs and that the strain's genome can evolve after passing through the digestive tract of pigs.