Search

Salmonella enterica serovar Heidelberg (S. Heidelberg) is a major foodborne pathogen colonizing poultry. The pathogen is associated with a significant number of foodborne outbreaks through contaminated poultry meat, including turkeys. Recently, multidrugresistant (MDR) strains of S. Heidelberg have emerged as a threat to human public health in the United States. The objective of this study was to determine the cecal colonization, dissemination to internal organs, and the potential for skeletal muscle deposition of an MDR S. Heidelberg isolate from the 2011 ground turkey outbreak in the United States after the experimental oral challenge of poults (young turkeys) and adult turkey hens. In the poult study, two separate experiments using day-old, straight-run, commercial hybrid converter poults were randomly assigned to five challenge groups (0, 10^2, 10^4, 10^6, 10^8 CFU groups; 12 poults/group; N D 60/experiment) and a week after, treatment groups were challenged separately with 0-, 2-, 4-, 6-, and 8- log10 CFU of S. Heidelberg orally. After 14 days post-challenge, the poults were euthanized, and samples were collected to determine MDR S. Heidelberg colonization in the cecum, dissemination to liver and spleen, and deposition in the thigh, drumstick, and breast muscles. A similar experimental design was followed for the adult turkey hens. In two separate experiments, 11-week-old commercial Hybrid Converter turkey hens (4 hens/group; N D 20/experiment) were challenged with MDR S. Heidelberg and on day 16 post-challenge, birds were euthanized and samples were collected to determine Salmonella populations in the samples. The results indicated that, in turkey poults, the recovery of MDR S. Heidelberg was highest in the cecum followed by spleen, liver, thigh, drumstick, and breast. All tested inoculum levels resulted in more than 3.5 log10 CFU/g colonization in the poult cecum. The cecal colonization, dissemination to internal organs, and tissue deposition of MDR S. Heidelberg were high in poults. The pathogen recovery from the cecum of adult turkey hens ranged from 37.5 to 62.5% in the challenge groups. The results signify the importance of controlling MDR S. Heidelberg in turkeys at the farm level to improve the safety of turkey products. [ABSTRACT FROM AUTHOR]

Extended-spectrum cephalosporin (ESC)-resistant Salmonella have been described at a low level in the EU, nevertheless the increasing importation of poultry meat could be an important source of epidemic strains carrying ESC resistance genes. This study evaluated ESC resistance and its genetic platform among Salmonella isolates from poultry meat products imported into Portugal as well as clonal relatedness of the isolates. All Salmonella isolates recovered from samples of fresh meat destined for import into the EU in the scope of Portuguese official border control (2014–2015) were studied. Antibiotic susceptibility and β-lactamase production was determined by disk diffusion/microdilution. Molecular studies included detection of genes encoding acquired AmpC and extended-spectrum β-lactamases, plasmid-mediated quinolone resistance and other antibiotic resistance genes by PCR/sequencing, and clonality by MLST and Xba I-PFGE. Plasmid characterisation was assessed by conjugation assays, replicon typing (PCR-PBRT/pMLST) and hybridisation experiments (I- Ceu I/S1-PFGE nuclease). Isolates belonged to Salmonella Heidelberg ( n  = 6; ST15/eBG26) and Salmonella Minnesota ( n  = 1; ST548/eBG77) and presented multidrug-resistant profiles, including to ESCs and/or fluoroquinolones. All but one carried bla CMY-2 , located on two epidemic plasmids, IncA/C (ST2, n  = 5) or transferable IncI1 (ST12, n  = 1). Salmonella Heidelberg was associated with five PFGE types, including one similar to an American epidemic clone. This study reveals imported poultry products as a source of uncommon and/or invasive ESC-resistant Salmonella strains in the EU. The increase of clinically relevant poultry-related serotypes in Europe must be taken into account in the current monitoring of antibiotic resistance trends and in re-evaluation of food regulations. [ABSTRACT FROM AUTHOR]

Salmonellosis ranks among the major diseases of commercial poultry, and its presence in poultry flocks is responsible for economic losses and risks related to public health. Vaccines are an important tool within integrated programmes to control salmonellosis. The purpose of this study was to assess cross-protection provided by the Poulvac® ST vaccine in the control ofSalmonellaHeidelberg in experimentally challenged 3- and 21-day-old birds. Eighty birds were identified and separated into four treatments (T1: vaccinated and challenged at 3 days of age, T2: unvaccinated and challenged at 3 days of age, T3: vaccinated and challenged at 21 days of age, and T4: unvaccinated and challenged at 21 days of age). The inoculum was produced from a Brazilian field strain of SH. At the end of the experiment, caecum and liver/spleen samples were collected for quantitative and qualitative analysis of SH, respectively. Analysis of the liver/spleen showed that Poulvac® ST significantly (P ≤ 0.05) reduced the percentage of SH positivity in the group challenged at 3 days of age, while in the group challenged at 21 days this difference was almost considered significant (P = 0.1818). On the other hand, there was no statistically significant difference in SH count in the caecum (CFU/g) in the group challenged at 3 days, but for the group challenged at 21 days the SH counts were significantly (P ≤ 0.05) lower in the vaccinated group when compared to the positive control. [ABSTRACT FROM AUTHOR]

Salmonella attached to the poultry skin is a major source of carcass contamination during processing. Once attached to the poultry skin, it is difficult to detach and inactivate Salmonella by commonly used antimicrobial agents since the pathogen is entrapped deeply in the feather follicles and the crevices on the skin. Essential oils could be natural, safe, and effective alternatives to synthetic antimicrobial agents during commercial and organic processing setup. The present study evaluated the efficacy of pimenta (Pimenta officinalis Lindl.) leaf essential oil (PEO), and its nanoemulsion in reducing Salmonella Heidelberg attachment on to turkey (Meleagris gallopavo) skin during simulated scalding (65°C) and chilling (4°C) steps in poultry processing. A multidrug resistant S. Heidelberg isolate from the 2011 ground turkey outbreak in the United States was used in the study. Results showed that PEO and the nanoemulsion resulted in significant reduction of S. Heidelberg attachment on turkey skin. Turkey skin samples treated with 1.0% PEO for 5 min resulted in >2 log10 CFU/sq. inch reduction of S. Heidelberg at 65 and 4°C, respectively (n = 6; P < 0.05). Similarly, skin samples treated with 1.0% pimenta nanoemulsion (PNE) for 5 min resulted in 1.5- and 1.8- log10 CFU/sq. inch reduction of S. Heidelberg at 65 and 4°C, respectively (n = 6; P < 0.05). In addition, PEO and PNE were effective in reducing S. Heidelberg on skin during short-term storage at 4 and 10°C (temperature abuse) (n = 6; P < 0.05). No Salmonella was detected in the dipping solution containing 0.5 or 1.0% PEO or PNE, whereas a substantial population of the pathogen survived in the control dipping solution. The results were validated using scanning electron -, and confocal - microscopy techniques. PEO or PNE could be utilized as an effective antimicrobial agent to reduce S. Heidelberg attachment to turkey skin during poultry processing.

Salmonella enterica is one of the most prevalent foodborne pathogens. The large quantity of serovar types results in the colonization of a large spectrum of hosts, with different environmental conditions and hazards. The aim of this study was to evaluate the differences in gene expression (bcsA and csgD) of Salmonella enterica serovars Heidelberg, Kentucky, and Enteritidis during biofilm formation using quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). Overall, there appeared to be differences in expression between the different serovars with high variation between strains. These data are important as they demonstrate considerable variability in gene expression between serovars and strains of poultry isolates of Salmonella enterica.

Salmonella attached to the poultry skin is a major source of carcass contamination during processing. Once attached to the poultry skin, it is difficult to detach and inactivate Salmonella by commonly used antimicrobial agents since the pathogen is entrapped deeply in the feather follicles and the crevices on the skin. Essential oils could be natural, safe, and effective alternatives to synthetic antimicrobial agents during commercial and organic processing setup. The present study evaluated the efficacy of pimenta (Pimenta officinalis Lindl.) leaf essential oil (PEO), and its nanoemulsion in reducing Salmonella Heidelberg attachment on to turkey (Meleagris gallopavo) skin during simulated scalding (65°C) and chilling (4&#176C) steps in poultry processing. A multidrug resistant S. Heidelberg isolate from the 2011 ground turkey outbreak in the United States was used in the study. Results showed that PEO and the nanoemulsion resulted in significant reduction of S. Heidelberg attachment on turkey skin. Turkey skin samples treated with 1.0% PEO for 5 min resulted in >2 log10 CFU/sq. inch reduction of S. Heidelberg at 65 and 4° C, respectively (n = 6; P < 0.05). Similarly, skin samples treated with 1.0% pimenta nanoemulsion (PNE) for 5 min resulted in 1.5- and 1.8- log10 CFU/sq. inch reduction of S. Heidelberg at 65 and 4° C, respectively (n = 6; P < 0.05). In addition, PEO and PNE were effective in reducing S. Heidelberg on skin during short-term storage at 4 and 10° C (temperature abuse) (n = 6; P < 0.05). No Salmonella was detected in the dipping solution containing 0.5 or 1.0% PEO or PNE, whereas a substantial population of the pathogen survived in the control dipping solution. The results were validated using scanning electron -, and confocal - microscopy techniques. PEO or PNE could be utilized as an effective antimicrobial agent to reduce S. Heidelberg attachment to turkey skin during poultry processing. [ABSTRACT FROM AUTHOR]

Copyright of Avian Diseases is the property of American Association of Avian Pathologists, Inc. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Contaminated eggs produced by infected commercial laying flocks are often implicated as sources of human infections with Salmonella Enteritidis, but Salmonella serovars Heidelberg and Typhimurium have also been associated with eggtransmitted illness. Contamination of the edible contents of eggs is a consequence of the colonization of reproductive tissues in systemically infected hens. In recent years, the animal welfare implications of diverse poultry housing and management systems have been vigorously debated, but the food safety significance of laying hen housing remains uncertain. The present study evaluated the effects of 2 different bird stocking densities on the invasion of internal organs by Salmonella serovars Heidelberg and Typhimurium in groups of experimentally infected laying hens housed in colony cages enriched with perching and nesting areas. Laying hens were distributed at 2 different stocking densities (648 and 973 cm²/bird) into colony cages and (along with a group housed in conventional cages at 648 cm²/bird) orally inoculated with doses of 107 cfu of 2-strain cocktails of either Salmonella Heidelberg or Salmonella Typhimurium. At 5 to 6 d postinoculation, hens were euthanized and samples of internal organs (cecum, liver, spleen, ovary, and oviduct) were removed for bacteriologic culturing. The overall frequency of Salmonella isolation from ceca after inoculation with strains of serovar Heidelberg (83.3%) was significantly (P < 0.001) greater than the corresponding value for strains of serovar Typhimurium (53.8%), whereas Salmonella was recovered significantly more often from both livers (85.2% vs. 53.7%; P < 0.0001) and spleens (78.7% vs. 56.5%; P = 0.0008) after inoculation with strains of serovar Typhimurium than strains of serovar Heidelberg. However, there were no significant differences (P > 0.05) between stocking densities or cage systems in the frequencies of isolation of either Salmonella serovar from any of the five sampled tissues. These results contrast with prior studies, which reported increased susceptibility to internal organ invasion by Salmonella Enteritidis among hens in conventional cages at higher stocking densities. [ABSTRACT FROM AUTHOR]

The microbiological risk of recycled litter depends on the efficacy of the management system applied to inactivate residual microorganisms and preserve the health of the successive broiler flock. This study aimed to assess the viability and infectivity of the Newcastle Disease Virus (NDV), Infectious Bursal Disease Virus (IBDV) and Salmonella Heidelberg in recycled litter exposed to different treatments. The litter was contaminated with microorganisms and submitted to the treatments (T): T1: shallow fermentation; T2: quicklime (calcium oxide); T3: shallow fermentation followed by addition of quicklime; T4: no treatment. Sentinel chicks housed on the treated litter showed that T1 and T3 inactivated residual IBDV. Analysis of the litter subjected to T1 also showed reduced levels of total enterobacteria. T2 was not able to reduce the microorganisms assessed and its association with T1 (T3) failed to enhance the effect of the treatment. NDV did not survive in the broiler litter, regardless of the treatment applied, and it was also not detected in the sentinel chicks. S. Heidelberg remained viable in the litter submitted to all studied treatments, being isolated from the sentinel chicks of all the experimental groups. The antimicrobial activity of T1 and T3 was associated to higher ammonia contents in the broiler litter. The results indicate that the shallow fermentation treatment is efficient for controlling residual IBDV and total enterobacteria in the recycled litter. [ABSTRACT FROM AUTHOR]

Salmonella enterica serovar Heidelberg ( S. Heidelberg) is one of the pathogens most frequently detected in recent years in food products from animals; here, we examine the acid tolerance of this strain. Mild acid (pH 5.5–5.0) induced a strong acid resistance in S. Heidelberg; the induced acid resistance improved within 0.5–6 h and resulted in > 95% cell survival following challenge in pH 3.0 medium. Addition of lysine or arginine to pH 2.5 acid challenge medium significantly improved the survival of S. Heidelberg; lysine induced the largest increase in survival. The lysine and arginine decarboxylase-related genes (i.e., cadA , cadB , adiA , and adiY ) are acid induced genes, and they play an important role in S. Heidelberg acid resistance. RT-PCR showed that the genes expression levels increased as acid adaptation pH decreased. The increased expression was maintained for at least 4 h during adaptation. Moreover, acid adaptation may have increased the production of cellulose and swimming in S. Heidelberg; pH 5.5 acid-adapted cells showed a red, dry, and rough morphotype on Congo red plates. The transcriptional levels of Enterotoxin gene ( stn ) increased three times following acid adaptation; however, the expression of Salmonella pathogenicity island 1 virulence genes significantly decreased. [ABSTRACT FROM AUTHOR]

Objetivou-se avaliar a ação de três princípios ativos rotineiramente utilizados na higienização de abatedouros avícolas frente a amostras de Salmonella Heidelberg isoladas em diferentes pontos da tecnologia de abate de um mesmo frigorífico. Foram testadas 20 amostras de S. Heidelberg (14 isoladas em 2005 e seis em 2009) frente a clorexidina (0,5%), amônia quaternária (0,5%) e ácido peracético (1%) nos tempos de contato de 5, 10, 15 e 20 minutos. Todas as amostras foram sensíveis ao ácido peracético 1% em todos os tempos testados. Observou-se que 100% das amostras isoladas em 2005 foram sensíveis a amônia quaternária enquanto que as isoladas em 2009 apresentaram 33% de resistência com 5 minutos de contato e 16,6% com 10 minutos de contato. Com relação à clorexidina, 25% dos isolados em 2005 mostraram-se resistentes após 5 minutos de contato enquanto que 33% das amostras isoladas em 2009 foram resistentes neste tempo e 17% no tempo de 10 minutos de contato. Pode-se concluir que o ácido peracético teve ação in vitro sobre as amostras isoladas em 2005 e 2009, enquanto que a clorexidina e a amônia quaternária tiveram sua ação reduzida frente às amostras de 2009, indicando a progressão da resistência bacteriana frente a estes sanitizantes e a necessidade de testes periódicos e rotação de princípios ativos nos programas de higienização dos frigoríficos.

This study aimed at determining the minimum time required for the penetration of Salmonella Heidelberg inside the eggs after contact with contaminated material. Recently-collected brown and white eggs from laying hens between 45-50 weeks of age, reared in a commercial poultry house, were artificially contaminated by contact with wood shavings moistened with liquid inoculum of Salmonella Heidelberg in stationary-growth phase (10³-10(4) CFU g-1). According to type (white or brown), eggs were distributed into three different groups, with four replicates each: negative control group (no artificial contamination), positive control group (analyzed externally immediately after contamination and internally after the maximum storage period of the test group) and test group. Eggs were stored at controlled environmental temperature varying from 25ºC to 30ºC. In the test group, eggs contents (yolk and albumen) were pooled and analyzed after 1:00, 1:30, 2:00, 2:30, 3:00, 3:30, and 4:00 hours after contamination for the presence of Salmonella Heidelberg in 25g of this pool. The experimental unit consisted of five eggs in each test. The analysis protocol included pre-enrichment, selective enrichment, plating on selective agar, and biochemical and serological tests. The results obtained were submitted to logistic regression, which indicated that the presence of Salmonella Heidelberg was verified after 2:16 h and 2:44 h of contact with white and brown eggs, respectively.

Salmonellaserovars, one of the leading contributors to foodborne illness and are especially problematic for foods that are not cooked before consumption, such as fresh produce. The shipping containers that are used to transport and store fresh produce may play a role in cross contamination and subsequent illnesses. However, methods for quantitatively attached cells are somewhat variable. The overall goal of this study was to compare conventional plating with molecular methods for quantitating attached representative strains forSalmonellaTyphimurium and Heidelberg on reusable plastic containers (RPC) coupons, respectively. We attachedSalmonella entericaserovar Typhimurium ATCC 14028 and serovar Heidelberg SL486 (parent and an antibiotic resistant marker strain) to plastic coupons (2.54 cm2) derived from previously used shipping containers by growing for 72 h in tryptic soy broth. The impact of the concentration of sanitizer on log reductions between unsanitized and sanitized coupons was evaluated by exposing attachedS. Typhimurium cells to 200 ppm and 200,000 ppm sodium hypochlorite (NaClO). Differences in sanitizer effectiveness between serovars were also evaluated with attachedS. Typhimurium compared to attachedS. Heidelberg populations after being exposed to 200 ppm peracetic acid (PAA). Treatment with NaClO caused an average of 2.73 ± 0.23 log CFU ofS. Typhimurium per coupon removed with treatment at 200 ppm while 3.36 ± 0.54 log CFU were removed at 200,000 ppm. Treatment with PAA caused an average of 2.62 ± 0.15 log CFU removed forS. Typhimurium and 1.41 ± 0.17 log CFU forS. Heidelberg (parent) and 1.61 ± 0.08 log CFU (marker). Lastly, scanning electron microscopy (SEM) was used to visualize cell attachment and coupon surface topography. SEM images showed that remaining attached cell populations were visible even after sanitizer application. Conventional plating and qPCR yielded similar levels of enumerated bacterial populations indicating a high concordance between the two methods. Therefore, qPCR could be used for the rapid quantification ofSalmonellaattached on RPC. [ABSTRACT FROM PUBLISHER]

Escalating product recalls as a consequence of Salmonella-contaminated poultry products have resulted in detrimental economic impacts in the poultry industry. One potential long-term alternative method to Salmonella prevention is genetic selection to improve innate resistance. This study evaluated the ex vivo effects of Salmonella Heidelberg (SH) on phagocytic and bactericidal leukocyte function in turkeys from six pedigree lines (A–F). Dayof- hatch poults (n = 48) were placed and raised in cages (2 birds/gender/genetic line/cage) to 35 d when heterophils and peripheral blood mononuclear cells (PBMCs) were extracted from males and females of each line. Cells were used in phagocytic and bactericidal assays to determine the ex vivo effects of SH on turkey leukocyte activity. Data were analyzed using the Fit Model platform in JMP Pro 10.0 (SAS Institute Inc.) with differences considered significant at P ≤ 0.05 and data reported as LS Means with SEM. Although genetic line had no significant effect on phagocytosis of SH by heterophils and PBMCs, cumulatively, female cells exhibited higher phagocytosis potential than those from males. The main effect of gender was significant on bactericidal activity of PBMCs when incubated at a 1:10 and 1:100 PBMC to SH ratio. Genetic line also had a significant effect on bactericidal activity of PBMCs with cells from line F exhibiting the best activity. These results suggest that gender had a marked cumulative effect on phagocytosis of SH by heterophils and PBMCs while both genetic line and gender had a prominent effect on bacterial killing of SH by turkey PBMCs. Once able to determine genetic markers associated with these immune responses to Salmonella, genetic selection for increased resistance may become feasible in turkeys. [ABSTRACT FROM AUTHOR]

Salmonella remains the second most common cause of gastrointestinal illness in the United States, European Union and other regions of the globe. In the food chain, poultry, especially turkeys and chickens, are considered significant contributors to salmonellosis in humans, as their intestinal tracts are often colonized with Salmonella. Due to changes in the prevalence of serovars, Salmonella Heidelberg (SH) has gained prominence, especially in the United States and Brazil, where outbreaks have occurred in recent years. The transmission of salmonella can occur by different routes, whether by infected breeding flocks, previously infected flocks, contaminated food or water. Thus, eliminating the bacteria from the installations can be a difficult task. In this sense, several countries have implemented salmonella control programs on poultry farms, in order to reduce its presence. Our study demonstrated that, despite the similarity between bird species and their rearing systems, turkeys are more sensitive to the Heidelberg serovar, especially in relation to microbiological contamination of internal organs and elimination of the agent into the environment. This information is extremely relevant for the poultry industry, enabling it to improve biosecurity programs in this species. [ABSTRACT FROM AUTHOR]

We have previously described that a strain of Salmonella Heidelberg with a hypermutator phenotype, B182, adhered strongly to HeLa cells. In this work, we showed that this hypermutator Salmonella strain invaded HeLa epithelial cells and induced cytoskeleton alteration. Those changes lead to HeLa cell death which was characteristic of apoptosis. For the first time, we showed that this hypermutator strain induced apoptosis associated with the activation of caspases 2, 9 and 3. Complementation of B182 strain showed a decrease in cells death induction. In the presence of other Salmonella Heidelberg with a normomutator phenotype, such as WT and SL486, cell death and caspase 3 were undetectable. These results suggested that early apoptosis and caspase 3 activation were specific to B182. Besides, B182 induced LDH release and caspase 3 activation in CaCo-2 and HCT116 cells. Heat-treated B182 and diffusible products failed to induce this phenotype. Epithelial cells treatment with cytochalasin D caused the inhibition of B182 internalisation and caspase 3 activation. These results showed that this cell death required active S . Heidelberg B182 protein synthesis and bacterial internalisation. However sipB and sopB , usually involved in apoptosis induced by Salmonella were not overexpressed in B182, contrary to fimA and fliC . Comparative genome analysis showed numerous mutations as in rpoS which would be more investigated. The role of the hypermutator phenotype might be suspected to be implicated in these specific features. This result expands our knowledge about strong mutators frequently found in bacterial organisms isolated from clinical specimens. [ABSTRACT FROM AUTHOR]

A stochastic model was used to estimate the annual number of human cases of ceftiofur-resistant Salmonella Heidelberg in Québec and Ontario (2003–2011) attributable to chicken consumption and human antimicrobial consumption, using Canadian surveillance data.A stochastic model was used to estimate the number of human cases of ceftiofur-resistant Salmonella enterica serovar Heidelberg in Québec and Ontario attributable to chicken consumption and excess cases attributable to human prior antimicrobial consumption. The annual mean incidence of S. Heidelberg (Québec/Ontario) decreased from 70/62 cases per 100 000 in 2004 to 29/30 cases per 100 000 in 2007 (Québec)/2008 (Ontario), increasing to 59/45 cases per 100 000 in 2011. The annual mean incidence of ceftiofur-resistant cases from chicken decreased from 8/7 cases per 100 000 in 2004 to 1/1 cases per 100 000 in 2007 (Québec)/2008 (Ontario), increasing to 7/5 cases per 100 000 in 2011. The annual mean total number of excess ceftiofur-resistant cases from chicken attributable to human prior antimicrobial consumption (Québec/Ontario) decreased from 71/123 in 2004 to 6/24 in 2007 (Québec)/2008 (Ontario), but increased to 62/91 in 2011. This model will support future work to determine the increased severity, mortality and healthcare costs for ceftiofur-resistant Salmonella Heidelberg infections. These results provide a basis for the evaluation of future public health interventions to address antimicrobial resistance. [ABSTRACT FROM AUTHOR]

Background Salmonella enterica subsp. enterica serovars Typhimurium (S. Typhimurium) and Heidelberg (S. Heidelberg) are major causes of foodborne salmonellosis, accounting for a fifth of all annual salmonellosis cases in the United States. Rapid, efficient and accurate methods for identification are required for routine surveillance and to track specific strains during outbreaks. We used Pulsed-field Gel Electrophoresis (PFGE) and a recently developed molecular subtyping approach termed CRISPR-MVLST that exploits the hypervariable nature of virulence genes and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) to subtype clinical S. Typhimurium and S. Heidelberg isolates. Results We analyzed a broad set of 175 S. Heidelberg and S. Typhimurium isolates collected over a five-year period. We identified 21 Heidelberg Sequence Types (HSTs) and 37 Typhimurium STs (TSTs) that were represented by 27 and 45 PFGE pulsotypes, respectively, and determined the discriminatory power of each method. Conclusions For S. Heidelberg, our data shows that combined typing by both CRISPR-MVLST and PFGE provided a discriminatory power of 0.9213. Importantly, CRISPR-MVLST was able to separate common PFGE patterns such as JF6X01.0022 into distinct STs, thus providing significantly greater discriminatory power. Conversely, we show that subtyping by either CRISPR-MVLST or PFGE independently provides a sufficient discriminatory power (0.9345 and 0.9456, respectively) for S. Typhimurium. Additionally, using isolates from two S. Typhimurium outbreaks, we demonstrate that CRISPR-MVLST provides excellent epidemiologic concordance. [ABSTRACT FROM AUTHOR]

This study describe the use of a combination of two recently proposed typing approaches, multiple amplification of prophage locus typing (MAPLT) and multiple-locus variable-number tandem-repeat analysis (MLVA) for subdividing within Salmonella enterica serovar Heidelberg ( S. Heidelberg). The combined typing method was compared with pulsed-field gel electrophoresis (PFGE) by Simpson's index of diversity (DI). PFGE was shown to have a DI = 0.84 and was poor at differentiation of the predominant PT1 (Phage Type 1) phenotype. In comparison, the combined MAPLT/MLVA method comprising 3 MLVA and 9 MAPLT primer pairs provided a higher differentiating ability DI = 0.92. More importantly, the combined methodology was found to be superior in the differentiation of the predominant PT1 isolates. In conclusion, this study demonstrated the potential of the rapid and simple amalgamated MAPLT/MLVA approach in determining transmission of isolates of clonal phage type groups from various environmental sources to humans. [ABSTRACT FROM AUTHOR]

Abstract: In this study, we investigated adherence and motility of the hypermutator Salmonella enterica Heidelberg B182 bovine strain related to a 12bp deletion in mutS. This mutator phenotype was associated with increased adherence to epithelial cells and with high expression of fimA as shown by real-time RT-PCR. Motility studies showed that fliC were up-regulated in the B182 strain, while fljA and fljB were down-regulated. In order to determine if mutated mutS is implicated in this genes expression, isogenic strains, derived from a WT strain, containing the 12bp deletion in mutS (Δ12bpmutS) or an inactivated mutS (ΔmutS) were generated. Δ12bpmutS and ΔmutS strains showed a spontaneous mutation rate similar to the environmental strain B182, but exhibited lower adherence capacity and fimA expression. In contrast to the fimbriae genes, in Δ12bpmutS, fliC expression was up-regulated, but fljA and fljB expression were decreased, as in the B182 strain. Only fljB expression was increased in ΔmutS mutants. Taken together, our data suggest that mutS alteration does not influence fimbriae expression but can impact flagella genes. [Copyright &y& Elsevier]

In the present study, a series of experiments was conducted to evaluate the ability of a commercial probiotic culture (FloraMax, IVS-Wynco LLC, Springdale, AR) to reduce Salmonella enterica serovar Heidelberg (SH) in chicks and turkey poults. In experiments 1 and 2, chicks were randomly assigned to treatment groups and then challenged via oral gavage with SH. Chicks were treated 1 h following SH challenge with the probiotic culture via oral gavage. At 24 and 72 h posttreatment, cecal tonsils and ceca were collected for recovery and enumeration of enteric Salmonella Heidelberg, respectively. In experiment 3, day-of-hatch turkeys were randomly assigned to treatment groups and then challenged via oral gavage with SH. Poults were treated 1 h following challenge with the probiotic via oral gavage. At 24 and 72 h post probiotic treatment, cecal tonsils and ceca were collected for recovery and enumeration of enteric SH, respectively. The probiotic culture significantly reduced the incidence of SH in cecal tonsils at both time points in chicks in both experiments (P < 0.05). These data demonstrate that administration of probiotic 1 h post SH challenge significantly reduced the incidence of SH recovery from cecal tonsils of chicks compared with controls 24 and 72 h following treatment. Similarly, probiotic treatment resulted in significant reductions in the concentrations of SH within the ceca in both experiments. Although similar significant results were observed at both 24 and 72 h in experiment 3, it was clear that poults were more susceptible to SH colonization than chicks. Overall, a Lacto bacillus-based probiotic significantly reduced Salmonella enterica serovar Heidelberg in chicks and turkey poults. [ABSTRACT FROM AUTHOR]

Abstract: Hypermutation is an important mechanism used by different Salmonella enterica subspecies enterica to regulate genetic stability in adaptation to changing environments, including antimicrobial treatments and industrial processes. Strong hypermutator strains generally contain a mutation in genes of the methyl mismatch repair (MMR) system and have mutation frequencies up to 1000-fold higher than wild type strains. The objectives of this study were to determine the distribution of mutation frequencies from a collection of 209 Salmonella strains, to genetically characterize a strong mutator, and to study MMR mutated protein–DNA binding interactions. Only one strain of S. Heidelberg was determined to have a hypermutator phenotype by virtue of its high mutation rate. Sequencing of genes of the MMR system showed a 12bp deletion in the mutS gene was present. The MMR mutated protein-DNA binding interactions were studied by bioanalysis, using the available crystal structure of a similar MutS protein from Escherichia coli. This analysis showed the small deletion in the Salmonella MutS was localized within the core domain. A retardation assay with MutS from hypermutable and wild type strains showed this mutation has no effect on MutS DNA binding. A better understanding of the genetic mechanisms of hypermutation will help to anticipate the behavior of hypermutator strains in various conditions. [Copyright &y& Elsevier]

Salmonella Heidelberg is one of the 3 most frequently isolated serovars from human Salmonella cases in Canada, and the fourth most commonly reported Salmonella serovar in human foodborne disease cases in the United States. Since 1962, Salmonella Heidelberg has been isolated and reported in poultry and poultry products in Brazil. The poultry industry has focused efforts on reducing salmonellae incidence in live production in an effort to reduce Salmonella in the processing plant. A better understanding of the initial infection in chicks could provide approaches to control Salmonella contamination. The objective of the present study was to evaluate 2 Salmonella Heidelberg strains that differed in the presence of virulence genes invA, agfA, and lpfA; antimicrobial resistance profiles; and epidemiologic profiles on aspects of pathogenicity and intestinal morphology. Newly hatched broiler chicks were inoculated with 2 strains (SH23 and SH35) of Salmonella Heidelberg and cecal morphometry, histopathology, electron microscopy, and bacterial counts in the liver and cecum were assessed. The SH23 and SH35 strains resulted in different changes in villi height and crypt depth and inflammatory cell infiltration in the cecum. The SH35 group had higher liver and cecum bacterial cell counts when compared with SH23 strains. [ABSTRACT FROM AUTHOR]

Although Salmonella deposition inside yolks is uncommon in naturally contaminated eggs, migration through the vitelline membrane into the nutrient-rich yolk contents could enable rapid bacterial multiplication. Egg refrigeration restricts both penetration and growth, but a recently proposed national Salmonella Enteritidis control program would allow unrefrigerated ambient temperature storage of eggs on farms for up to 36 h. The present study used an in vitro egg contamination model to assess the ability of small numbers of 4 Salmonella Enteritidis strains and 4 Salmonella Heidelberg strains to penetrate the vitelline membrane and multiply inside yolks during 36 h of storage at either 20 or 30°C. After inoculation onto the exterior surface of the vitelline membrane, all 8 Salmonella strains penetrated to the yolk contents (at a mean frequency of 45.1%), and most strains grew to significantly higher levels (with a mean log10 bacterial concentration of 2.2 cfu/mL) during incubation at 30°C. Significant differences in penetration frequency and yolk multiplication were observed between individual strains and between serotypes (Salmonella Enteritidis > Salmonella Heidelberg for both parameters). Penetration and multiplication were significantly less frequent during incubation at 20°C. These results demonstrate that controlling ambient temperatures during prerefrigeration storage may be an important adjunct to prompt refrigeration for limiting Salmonella growth in eggs and thereby for preventing egg-transmitted human illness. [ABSTRACT FROM AUTHOR]

The increase of multidrug-resistant pathogens of human and animal origins is a major public health concern. For a better understanding of the health consequences of multidrug-resistant bacteria transmitted from animal products to humans, the host interaction of zoonotic Salmonella isolates along with other pathogenic and commensal bacteria was evaluated using a human intestinal Caco-2 cell system. Multidrug-resistant S. Agona, S. Heidelberg, and S. Typhimurium possessed plasmid-mediated class 1 integrons. The S. Typhimurium DT104 isolate from ground beef showed the well-known genotypic and phenotypic resistance characteristics of the species, and contained the chromosomally located class 1 integron. Among the multidrug-resistant Salmonella isolates, the S. Heidelberg 219 had the highest invasion number at 1.0 × 104 CFU/mL, followed by the S. Typhimurium DT104 isolate at 7.7 × 103 CFU/mL. Listeria monocytogenes was the best performer among the tested species in invading the Caco-2 cell. Multidrug-resistant opportunistic pathogens Klebsiella pneumoniae and Pseudomonas aeruginosa were also able to invade the cells. The invasion of S. Heidelberg 219, S. Typhimurium DT104, L. monocytogenes, K. pneumoniae, and P. aeruginosa into the Caco-2 cells was not affected even in the presence of commensal E. coli. During the intracellular growth of S. Heidelberg 219, S. Typhimurium DT104, and L. monocytogenes, the bacterial counts increased 2 log cycles in 9 h in the Caco-2 cells. Therefore, these strains could rapidly proliferate after their invasion into the cells. [ABSTRACT FROM AUTHOR]