Salmonella Isolations from Snakes and Other Reptiles

Bacteriological examination of faeces and swabs from fourteen species of reptiles predominantly constrictors, yielded six serotypes of Salmonella as well as Arizona hinshawii, Proteus species, Citrobacter freundii and Enterobacter species. Organisms isolated were identified using conventional bacteriological methods and yielded two serotypes of salmonellae not previously identified in Canada: Salmonella nima and Salmonella 58:a.     

6-hydroxydopamine-mediated release of norepinephrine increases faecal excretion of Salmonella enterica serovar Typhimurium in pigs

Salmonella enterica serovar Typhimurium is an animal and zoonotic pathogen of worldwide importance. In pigs, transport and social stress are associated with reactivation and spread of Salmonella Typhimurium infection. The stress-related catecholamine norepinephrine (NE) has been reported to activate growth and virulence factor expression in Salmonella; however the extent to which NE contributes to stress-associated salmonellosis is unclear. We studied the impact of releasing NE from endogenous stores during Salmonella Typhimurium infection of pigs by administration of 6-hydroxydopamine (6-OHDA), which selectively destroys noradrenergic nerve terminals. Treatment of pigs with 6-OHDA 7 or 16 days post-oral inoculation with Salmonella Typhimurium produced elevated plasma NE levels and transiently, but significantly, increased faecal excretion of the challenge strain. Oral administration of NE to Salmonella Typhimurium-infected pigs also transiently and significantly increased shedding; however pre-culture of the bacteria with NE did not alter the outcome of infection. Salmonella has been proposed to sense and respond to NE via a homologue of the adrenergic sensor kinase QseC. A ΔqseC mutant of Salmonella Typhimurium was consistently excreted in lower numbers than the parent strain post-oral inoculation of pigs, though not significantly so. 6-OHDA treatment of pigs infected with the ΔqseC mutant also increased faecal excretion of the mutant strain, albeit to a lesser extent than observed upon 6-OHDA treatment of pigs infected with the parent strain. Our data support the notion that stress-related catecholamines modulate the interaction of enteric bacterial pathogens with their hosts.     

Incidence of Salmonella Contamination in Broiler Chickens in Saskatchewan

The incidence of Salmonella contamination in ten Saskatchewan broiler flocks varying in size from 6 200 to 14 000 was investigated from February, 1977 to April, 1979. Prior to the initial chick placement, brooding equipment, feed, water and fresh litter samples were found to be free of Salmonellae. Samples obtained from the clean and disinfected processing plant equipment before the commencement of daily operation were negative except the isolation for Salmonella anatum from the fingers of the defeathering machine in flock 4. There was no evidence of Salmonella contamination in flocks 5, 6, 8 and 10. The incidence of Salmonella was lower when cloacal swabs were taken from day old chicks fasted for 48 hours than for the same groups of chicks when carcasses were blended in nutrient broth (flocks 7 and 9). The blending of such chicks appears to be a more critical test. The serotypes isolated from eviscerated birds were the same as those isolated from used litter samples. Salmonella saintpaul was isolated from a water sample at 53 days in flock 1 and the same serotype was recovered from the intestinal contents and skin of eviscerated birds. Salmonella typhimurium was recovered from the eviscerated birds and neck samples in flock 3. In flock 4, S. saintpaul and S. anatum were isolated from 13% of the eviscerated birds sampled. Salmonella thompson, Salmonella agona and Salmonella heidelberg were recovered from 61%, 5% and 1%, respectively, of the processed carcasses sampled in flock 7.     

Persistent shedding of Salmonella enteritidis from the udder of a cow

Salmonella enteritidis phagetype 8 was isolated from ill humans, milk-line filters, milk from a bulk tank, and milk from the right hind quarter of a five-year-old Holstein cow on a dairy farm in southern Alberta. The affected animal was removed from the herd and continued to shed S. enteritidis from this quarter during a seven-month interval in an isolation facility. Milk from the affected quarter was visually normal, and no other pathogen was isolated from the udder during the investigation. After removal of the infected cow from the herd, milk from the bulk tank was culturally negative for Salmonella sp. during the succeeding 15 months.     

Salmonella and antimicrobial resistance in broilers: A review

Salmonella enterica is a zoonotic pathogen which can readily pass from animal to man through the consumption of contaminated food. The prevalence of Salmonella enterica associated with poultry and poultry meat products has been well-documented and this prevalence has both public health and economic implications. The estimated total cost for nontyphoidal Salmonella is in excess of 14 billion dollars/year in the United States alone. Almost 41,930 cases of nontyphoidal foodborne salmonellosis are confirmed annually with an estimated total number of 1 million cases of foodborne salmonellosis not reported. The emergence of antimicrobial resistant Salmonella recovered from meat products has heightened concerns regarding antimicrobial use in food animal production. This review will cover the history and taxonmy of Salmonella enterica, Salmonella in poultry and poultry products, colonization factors, transmission, detection and characterization, antibiotics, antimicrobial resistance, mechanisms of resistance in Salmonella by class, transmission of antimicrobial resistance, and the global implications of antimicrobial resistance.     

A temporal study of Salmonella serovars from environmental samples from poultry breeder flocks in Ontario between 1998 and 2008

A temporal study was carried out to determine Salmonella prevalence, trends, major serovars, and their clusters from environmental samples, in poultry breeder flocks in Ontario between January 1998 and December 2008. Surveillance data were obtained from the Ontario Hatchery and Supply Flock Policy. Logistic regression with a random effect for flock was used to identify factors [poultry type, year (trend) and season] associated with the prevalence of Salmonella. A cluster detection test was used to identify clusters of common serovars. The period prevalence of Salmonella was 47.4% in broiler-breeder, 25.7% in layer-breeder, and 19.6% in turkey-breeder flocks. The overall trend in the prevalence of Salmonella was decreasing for all breeder types, due primarily to decreasing trends of Salmonella Heidelberg. The seasonal effects varied by year with the highest probability of Salmonella occurring in different seasons. The 4 most common serovars identified were Salmonella Heidelberg, Kentucky, Hadar, and Typhimurium in broiler-breeders; Salmonella Heidelberg, Brandenburg, Thompson, and Typhimurium in layer-breeders; and Salmonella Heidelberg, Saintpaul, Brandenburg, and Muenster in turkey-breeders. Salmonella Enteritidis was infrequently isolated in all poultry breeder types. Temporal clusters of different serovars were identified in all poultry breeder types. Clusters of Salmonella Heidelberg, Typhimurium, and Hadar from environmental samples from breeder flocks were detected during a similar period to clusters from hatchery fluff samples from the same population. Therefore, interventions at the breeder flock-level might help to reduce transmission of Salmonella from breeder flocks to hatcheries and possibly, to lower levels of the poultry production chain.     

Isolation of scFv fragments specific to OmpD of Salmonella Typhimurium

Pork meat is one of the major sources for human infections with Salmonella enterica subspecies enterica serovars. Further, zoonoses caused by S. enterica subspecies enterica serovars are responsible for substantial economical losses in industrial countries. Quick and reliable detection of this infection is urgently needed to improve consumer security. Due to its capability to identify infections independent of the species, a competitive ELISA is the preferable method for the detection of anti-Salmonella antibodies in serum. Recombinant antibody fragments (scFvs) were isolated from the naive human antibody gene library HAL7 by phage display. Recombinant produced outer membrane protein D (OmpD) of Salmonella Typhimurium was used as antigen. The characterization of the isolated single chain Fv (scFv) antibodies was done by enzyme-linked immunosorbent assay (ELISA), immunoblot, sequencing, epitope mapping and size exclusion chromatography (SEC). The detection of anti-OmpD IgGs in swine sera by competitive ELISA was shown in a proof of principle concept. Furthermore, the developed competitive ELISA would be compatible to a recently published DIVA vaccine, allow to distinguish between infected and vaccinated pigs.     

Comparison of bacterial enriched-broth culture, enzyme linked immunosorbent assay, and broth culture-polymerase chain reaction techniques for identifying asymptomatic infections with Salmonella in swine

A polymerase chain reaction (PCR) assay was combined with a broth-culture enrichment system to detect Salmonella shed in feces from subclinically infected swine. The effectiveness of the broth culture-polymerase chain reaction (BC-PCR) assay to identify pigs shedding Salmonella in feces was compared with a microbiological culture and a commercial enzyme linked immunosorbent assay (ELISA) kit to detect Salmonella-specific serum antibody. A total of 67 pigs were tested by each of the 3 methodologies. Forty-one pigs tested positive for Salmonella by BC-PCR and ELISA identified 6 positives and 23 suspicious samples. It was shown that the BC-PCR assay is a rapid diagnostic tool for detecting of Salmonella shed by asymptomatic swine compared with current diagnostic technologies.     

Prevention of Salmonella contamination of finished soybean meal used for animal feed by a Norwegian production plant despite frequent Salmonella contamination of raw soy beans, 1994–2012

Background

Salmonella contaminated animal feed is a major source for introducing Salmonella into the animal derived food chain. Because soybeans frequently are contaminated with Salmonella, soybean meal used as animal feed material, a by-product of a “crushing plant” which produces oil from soybeans, can be important source of Salmonella in the animal feed.We report the successful control of Salmonella from 1994 to 2012 in a Norwegian crushing plant producing soybean meal from imported soy beans. The results are based on an officially supervised HACCP based program including annual testing of around 4000 samples.

Results

During the 19-year period, 34% of samples collected during unloading of ships delivering soybeans yielded Salmonella; the proportion of samples from ships that yielded Salmonella varied from 12-62% each year. Dust samples from all shiploads from South America yielded Salmonella. In total 94 serovars of Salmonella were isolated, including nine (90%) of the EU 2012 top ten serovars isolated from clinical cases of salmonellosis in humans, including major animal pathogenic serovars like Spp. Typhimurium and Enteritidis.The effectiveness of the HACCP based control was indicated by a low prevalence of Salmonella contamination in the clean area of the plant, which is considered to be the main reason for the successful prevention of Salmonella in the end product. Despite extensive testing, no sample from the finished soybean meal product was found to be Salmonella contaminated.

Conclusions

This study shows that a HAACP-based control program in a soybean crushing plant can produce Salmonella free soybean meal despite frequent Salmonella contamination of raw soybeans. That approach is suggested as an effective way to minimize the risk of Salmonella exposure of the animal feed mills and contamination of the subsequent animal feed chain.     

Efficacy of Eugenol Against a Salmonella enterica serovar Enteritidis Experimental Infection in Commercial Layers in Production

An experimental study (15-wk-old ISA Brown pullets) was conducted to establish the efficacy of the essential oil of Eugenia caryophyllata against Salmonella enterica serovar Enteritidis. The trial was composed of 4 groups. Pullets in groups 3 and 4 were fed with a commercial compound feed, and pullets in groups 1 and 2 were fed with the same feed plus the aromatic product at the dose of 250 g/Tm. At 19 wk old, the pullets in groups 1 and 3 were infected individually with an inoculum of 3.2 ± 0.8 × 10⁷ cfu of Salmonella enterica serovar Enteritidis/ pullet. During the postinoculation period, samples of feces and eggs were cultured, and pullets were killed 30 d postinoculation. The aromatic product containing eugenol seems to aid in the cleaning of the intestinal and systemic infections, and it also plays an important role in the control of Salmonella cross contamination in eggs.     

A report of an outbreak of Salmonella oranienburg in a hybrid mouse colony

Mice in a hybrid breeding colony developed symptoms compatible with a diagnosis of salmonellosis. Salmonella oranienburg was recovered from feces by conventional bacteriological techniques. Active salmonellosis appeared to be precipitated by the stress of pregnancy among breeder female BALB/c mice. Necropsy examination of the BALB/c breeder females revealed diarrhea, roughened hair coats, enlarged spleens, white foci in the liver, and skin abscesses. S. oranienburg was recovered from the spleen, cecum, oropharynx, uterus, and skin abscesses of affected mice. DBA/2N breeder males were asymptomatic and S. oranienburg was recovered from the cecum only. Breeding productivity indices dropped to unacceptable levels in rooms containing infected animals. Relatively normal production was observed among uninfected animals of the same strains in adjacent rooms, thus demonstrating the detrimental effect S. oranienburg had on this breeder colony.     

The consequence of low mannose-binding lectin plasma concentration in relation to susceptibility to Salmonella Infantis in chickens

Mannose-binding lectin (MBL) is a key protein in innate immunity. MBL binds to carbohydrates on the surface of pathogens, where it initiates complement activation via the lectin-dependent pathway or facilitates opsonophagocytosis. In vitro studies have shown that human MBL is able to bind to Salmonella, but knowledge in relation to chicken MBL and Salmonella is lacking. In order to study this relation day-old chickens from two selected lines L10H and L10L, differing in MBL serum concentration, were either orally infected with S. Infantis (S.123443) or kept as non-infected controls. The differences between healthy L10H and L10L chicken sublines were more profound than differences caused by the S. Infantis infection. The average daily body weight was higher for L10H than for L10L, regardless of infection, indicating beneficial effects of MBL selection on growth. Salmonella was detected in cloacal swabs and the number of Salmonella positive chickens during the experiment was significantly higher in L10L than L10H, indicating that MBL may affect the magnitude of Salmonella colonisation in day-old chickens. MBL expression was determined in ceca tissue by real-time RT-PCR. L10H chickens showed a significantly higher relative expression than L10L at days 1 and 41 pi, regardless of infection. Finally, flow cytometric analysis of whole blood from infected chickens showed that L10H had a significantly higher count of all assessed leucocyte subsets on day 5 pi, and also a higher count of monocytes on day 12 pi than L10L. No difference was observed between infected and non-infected L10L chicken.     

Use of Bacitracin and Roxarsone to Reduce Salmonella Heidelberg Shedding Following a Necrotic Enteritis Challenge Model

This study was performed to determine if the enteric disease necrotic enteritis results in higher shedding of Salmonella. Fifty percent of the chicks per pen were challenged with a naladixic acid-resistant Salmonella Heidelberg. A necrotic enteritis challenge model using a coccidia vaccine to induce intestinal epithelium damage was used to demonstrate that neither the disease necrotic enteritis nor the control of that disease with the antibiotic bacitracin methylene disalycilate had an effect on the incidence of Salmonella Heidelberg. However, the feed additive 3-nitro-4-hydroxyphenylarsonic acid, which controls the cecal coccidia Eimeria tenella, significantly reduced the amount of Salmonella Heidelberg in the environment of the chick as measured by dragswabs and also reduced the number of ceca positive in the contact chicks.     

Environmental Salmonella in Agricultural Fair Poultry Exhibits in Colorado

Salmonella enterica is a common zoonotic pathogen in humans. Transmission typically occurs through consumption of contaminated food products or contact with infected animals, including poultry or their environment. The objective of this study was to estimate the frequency of Salmonella contamination in the environment in poultry exhibits at agricultural fairs. Samples were collected from cages, feed, floors and tables in the exhibit and cultured for Salmonella. At least one environmental sample was positive for Salmonella in 10 of 11 fairs (91%), and Salmonella was isolated from 28 of 55 environmental samples (50.9%). Eleven different serotypes were detected. Results of this study demonstrate that environmental surfaces at agricultural fairs can be contaminated with Salmonella and could potentially serve as a route of transmission to bird owners and the general public. Poultry owners and the general public should be educated about the risks of Salmonella infection from the poultry exhibit environment. Agricultural fairs should consider instituting policies and practices to improve hygiene and mitigate the risk of zoonotic salmonellosis.     

Prevalence and antimicrobial susceptibility of Salmonella isolates in Pekin ducks from South Korea

An investigation was carried out to determine the prevalence and antibiotic resistance of Salmonella serotypes at South Korean duck farms. A total of 7119 samples collected from 72 duck farms in five provinces were examined from 2011 to 2012. The overall prevalence of Salmonella serotypes was 43.4% (69/159) in duck flocks from 65.2% (47/72) of the duck farms. Eighty-five strains were isolated from 69 duck flocks. Three serotypes of Salmonella enterica were identified such as S. Typhimurium (39/85), S. Enteritidis (44/85), and S. London (2/85). The prevalence of Salmonella infection decreased significantly in 3-week-old ducks compared to that in 1-week-old ducks (P < 0.05). All isolates except one were resistant to at least one antimicrobial and 27% of the isolates were resistant to 5–16 antimicrobials. Our findings provide baseline information on the degree of Salmonella infection and distribution of Salmonella serotypes in ducks and indicate that ducks should be considered an important source of foodborne pathogens.     

Induction of seroconversion and persistence of Salmonella Typhimurium in pigs are strain dependent

Foodborne salmonellosis is one of the most important bacterial zoonotic diseases worldwide. Salmonella Typhimurium is the serovar most frequently isolated from persistently infected slaughter pigs in Europe. Salmonella Typhimurium pathogenesis is host species specific. In addition, differences in in vitro behaviour of Salmonella Typhimurium strains have also been described, which may be reflected by a different course of infection within a host species. We compared the course of a Salmonella Typhimurium infection in pigs, using two Salmonella Typhimurium strains that were able to interfere with MHC II expression on porcine macrophages to a different extent in vitro. After experimental inoculation, blood and faecal samples from all pigs were collected at regular time points. At 40 days post inoculation (pi), animals were euthanized and tissue samples were bacteriologically analysed. The proportion of serologically positive piglets at 33 days pi was significantly higher in pigs that were inoculated with the strain that did not downregulate MHC II expression in vitro. Furthermore, this strain was less frequently shed and isolated in lower numbers from tonsils and ileocaecal lymph nodes than the strain that was able to markedly downregulate MHC II expression in vitro. We thus found that the delayed onset of seroconversion after oral inoculation of piglets with a particular Salmonella Typhimurium strain coincided with higher faecal shedding and increased persistence. Strain specific differences in Salmonella pathogenesis might thus have repercussions on the serological detection of Salmonella Typhimurium infections in pigs.     

Salmonella enterica Typhimurium infection causes metabolic changes in chicken muscle involving AMPK,fatty acid and insulin/mTOR signaling

Salmonella enterica serovar Typhimurium (Salmonella Typhimurium) infection of chickens that are more than a few days old results in asymptomatic cecal colonization with persistent shedding of bacteria. We hypothesized that while the bacterium colonizes and persists locally in the cecum it has systemic effects, including changes to metabolic pathways of skeletal muscle, influencing the physiology of the avian host. Using species-specific peptide arrays to perform kinome analysis on metabolic signaling pathways in skeletal muscle of Salmonella Typhimurium infected chickens, we have observed key metabolic changes that affected fatty acid and glucose metabolism through the 5''-adenosine monophosphate-activated protein kinase (AMPK) and the insulin/mammalian target of rapamycin (mTOR) signaling pathway. Over a three week time course of infection, we observed changes in the phosphorylation state of the AMPK protein, and proteins up and down the pathway. In addition, changes to a large subset of the protein intermediates of the insulin/mTOR pathway in the skeletal muscle were altered by infection. These changes occur in pathways with direct effects on fatty acid and glucose metabolism. This is the first report of significant cellular metabolic changes occurring systemically in chicken due to a Salmonella infection. These results have implications not only for animal production and health but also for the understanding of how Salmonella infection in the intestine can have widespread, systemic effects on the metabolism of chickens without disease-like symptoms.     

A rabbit model of non-typhoidal Salmonella bacteremia

Bacteremia is an important cause of morbidity and mortality in humans. In this study, we focused on the development of an animal model of bacteremia induced by non-typhoidal Salmonella. New Zealand White rabbits were inoculated with a human isolate of non-typhoidal Salmonella strain CVD J73 via the intra-peritoneal route. Blood samples were collected at specific time points and at euthanasia from infected rabbits. Additionally, tissue samples from the heart, lungs, spleen, gastrointestinal tract, liver and kidneys were obtained at euthanasia. All experimentally infected rabbits displayed clinical signs of disease (fever, dehydration, weight loss and lethargy). Tissues collected at necropsy from the animals exhibited histopathological changes indicative of bacteremia. Non-typhoidal Salmonella bacteria were detected in the blood and tissue samples of infected rabbits by microbiological culture and real-time PCR assays. The development of this animal model of bacteremia could prove to be a useful tool for studying how non-typhoidal Salmonella infections disseminate and spread in humans.     

Characterization of Salmonella enterica serovar Typhimurium and Salmonella enterica serovar 4,[5],12:i:- isolates from pigs presenting with diarrhea in Korea

Between 2011 and 2012, a total of 896 pig fecal samples were collected from nine provinces in Korea, and 50 salmonella enterica susp. enterica serovar Typhimurium (S. Typhimurium) was isolated. The characteristics of the 50 strains were analyzed, and 4 strains were identified as Salmonella enterica subsp. enterica serovar 4,[5],12:i:-. Salmonella 4,[5],12:i:- could not be distinguished from S. Typhimurium through phage typing, antimicrobial resistance testing or multiple-locus variable-number tandem repeat analysis (MLVA). However, among the four Salmonella 4,[5],12:i:- strains, one (KVCC-BA1400078) was identified as a Salmonella 4,[5],12:i:- clone isolated from humans in the United States, and another (KVCC-BA1400080) was identified as DT193, which has been primarily isolated from humans and animals in European countries. The presence of Salmonella 4,[5],12:i:- in Korea poses a significant threat of horizontal transfer between pigs and humans.     

Mutations in the Salmonella enterica serovar Choleraesuis cAMP-receptor protein gene lead to functional defects in the SPI-1 Type III secretion system

Salmonella enterica serovar Choleraesuis (Salmonella Choleraesuis) causes a lethal systemic infection (salmonellosis) in swine. Live attenuated Salmonella Choleraesuis vaccines are effective in preventing the disease, and isolates of Salmonella Choleraesuis with mutations in the cAMP-receptor protein (CRP) gene (Salmonella Choleraesuis ∆crp) are the most widely used, although the basis of the attenuation remains unclear. The objective of this study was to determine if the attenuated phenotype of Salmonella Choleraesuis ∆crp was due to alterations in susceptibility to gastrointestinal factors such as pH and bile salts, ability to colonize or invade the intestine, or cytotoxicity for macrophages. Compared with the parental strain, the survival rate of Salmonella Choleraesuis ∆crp at low pH or in the presence of bile salts was higher, while the ability of the mutant to invade intestinal epithelia was significantly decreased. In examining the role of CRP on the secretory function of the Salmonella pathogenicity island 1 (SPI-1) encoded type III secretion system (T3SS), it was shown that Salmonella Choleraesuis ∆crp was unable to secrete the SPI-1 T3SS effector proteins, SopB and SipB, which play a role in Salmonella intestinal invasiveness and macrophage cytotoxicity, respectively. In addition, caspase-1 dependent cytotoxicity for macrophages was significantly reduced in Salmonella Choleraesuis ∆crp. Collectively, this study demonstrates that the CRP affects the secretory function of SPI-1 T3SS and the resulting ability to invade the host intestinal epithelium, which is a critical element in the pathogenesis of Salmonella Choleraesuis.