A SYBR Green™ I-based real-time multiplexed PCR assay was developed targeting invA and spvB for the detection of Salmonella strains in shellfish after both hns and invA genes were identified in all Salmonella strains. Simultaneously, the 16S rRNA gene was used as a PCR internal amplification control (IAC). All 89 Salmonella strains tested in this study exhibited amplification of invA, whereas only 21 (23. 6 %) were PCR positive for spvB. The sensitivity of detection of all three targeted genes was 1 ng, which is equivalent to approximately 105 colony-forming unit (CFU) of Salmonella enterica. The analysis showed specific PCR products that were identified by reproducible melt temperature profiles (invA, 84. 27 ± 1. 7 °C; spvB, 88. 76 ± 1. 0 °C; and 16S rRNA gene, 87. 16 ± 0. 8 °C). The sensitivity of detection was 10 pg purified DNA (invA) or 105 CFU in 1 mL pure culture of S. enterica ATCC 14028. The above molecular detection method for Salmonella strains was successfully applied to the oyster homogenates (food matrix). An initial inoculum of 106 and 102 CFU Salmonella in 1 ml seeded oyster tissue homogenate was detected by multiplexed PCR for all three genes after 5 and 24 h of enrichment, respectively. Natural oysters isolated from Gulf of Mexico during the winter months exhibited negative PCR amplification results suggesting the absence of Salmonella. In contrast to conventional PCR, real-time multiplex PCR assay developed in this study is rapid and sensitive and will help Interstate Shellfish Sanitation Conference undertake appropriate measures to monitor Salmonella in oysters, thereby preventing disease outbreaks and consequently protecting consumer health. © 2012 Springer Science+Business Media, LLC.
Full Text Available A PCR protocol was developed to identify strains of Salmonella from different sources using a set of “primers” PCR specific to Salmonella , for the recognition of 119 bp invA gene sequence. The results met the maxims of selectivity: inclusiveness, all strains of Salmonella presented positive signal for the gene, exclusivity since no strain Salmonella ( Shigella, E. coli, E. coli O157: H7 showed no signal positive robustness because in the 14 strains of Salmonella expected signal was obtained in a total of four repetitions with equal working conditions in four non-consecutive days. Also it was reproducible, results were identical by modifying the reaction conditions. Comparing PCR with conventional culture, using non- selective pre-enrichment in peptone water and selective enrichment in selenite-cystine broth, it was found that the PCR was 100% sensitive and specific, with 100% of inclusiveness, 100% exclusive and 100% of both positive and negative predictive value.
Swamy, S C; Barnhart, H M; Lee, M.D.; Dreesen, D W
The presence of two virulence foci, invA and spvC, in Salmonella isolates obtained from poultry, wastewater, and human sources was determined. All isolates (n = 245) were positive for the invA gene sequence. Differences in degree of invasiveness were apparent with the Madin Darby canine kidney cell line, as only 79 of 159 randomly selected isolates (49.7%) tested were invasive at > 0.1% of the inoculum. 25% were invasive between 0.1 and 1.0% of the inoculum, and 24.5% were invasive at > 1.0% ...
González-Escalona, Narjol; Hammack, Thomas S; Russell, Mindi; Jacobson, Andrew P; De Jesús, Antonio J; Brown, Eric W; Lampel, Keith A
Salmonella enterica contamination in foods is a significant concern for public health. When DNA detection methods are used for analysis of foods, one of the major concerns is false-positive results from the detection of dead cells. To circumvent this crucial issue, a TaqMan quantitative real-time RT-PCR (qRT-PCR) assay with an RNA internal control was developed. invA RNA standards were used to determine the detection limit of this assay as well as to determine invA mRNA levels in mid-exponential-, late-exponential-, and stationary-phase cells. This assay has a detection limit of 40 copies of invA mRNA per reaction. The levels of invA mRNA in mid-exponential-, late-exponential-, and stationary-phase S. enterica cells was approximately 1 copy per 3 CFU, 1 copy per CFU, and 4 copies per 10(3) CFU, respectively. Spinach, tomatoes, jalapeno peppers, and serrano peppers were artificially contaminated with four different Salmonella serovars at levels of 10(5) and less than 10 CFU. These foods were analyzed with qRT-PCR and with the FDA's Bacteriological Analytical Manual Salmonella culture method (W. A. Andrews and T. S. Hammack, in G. J. Jackson et al., ed., Bacteriological analytical manual online, http://www.cfsan.fda.gov/ approximately ebam/bam-5.html, 2007). Comparable results were obtained by both methods. Only live Salmonella cells could be detected by this qRT-PCR assay, thus avoiding the dangers of false-positive results from nonviable cells. False negatives (inhibition of the PCR) were also ruled out through the use of an RNA internal control. This assay allows for the fast and accurate detection of viable Salmonella spp. in spinach, tomatoes, and in both jalapeno and serrano peppers. PMID:19376910
González-Escalona, Narjol; Hammack, Thomas S.; Russell, Mindi; Jacobson, Andrew P.; De Jesús, Antonio J.; Brown, Eric W.; Lampel, Keith A.
Salmonella enterica contamination in foods is a significant concern for public health. When DNA detection methods are used for analysis of foods, one of the major concerns is false-positive results from the detection of dead cells. To circumvent this crucial issue, a TaqMan quantitative real-time RT-PCR (qRT-PCR) assay with an RNA internal control was developed. invA RNA standards were used to determine the detection limit of this assay as well as to determine invA mRNA levels in mid-exponent...
González-Escalona, Narjol; Hammack, Thomas S.; Russell, Mindi; Jacobson, Andrew P.; De Jesús, Antonio J.; Brown, Eric W.; Lampel, Keith A.
Salmonella enterica contamination in foods is a significant concern for public health. When DNA detection methods are used for analysis of foods, one of the major concerns is false-positive results from the detection of dead cells. To circumvent this crucial issue, a TaqMan quantitative real-time RT-PCR (qRT-PCR) assay with an RNA internal control was developed. invA RNA standards were used to determine the detection limit of this assay as well as to determine invA mRNA levels in mid-exponential-, late-exponential-, and stationary-phase cells. This assay has a detection limit of 40 copies of invA mRNA per reaction. The levels of invA mRNA in mid-exponential-, late-exponential-, and stationary-phase S. enterica cells was approximately 1 copy per 3 CFU, 1 copy per CFU, and 4 copies per 103 CFU, respectively. Spinach, tomatoes, jalapeno peppers, and serrano peppers were artificially contaminated with four different Salmonella serovars at levels of 105 and less than 10 CFU. These foods were analyzed with qRT-PCR and with the FDA's Bacteriological Analytical Manual Salmonella culture method (W. A. Andrews and T. S. Hammack, in G. J. Jackson et al., ed., Bacteriological analytical manual online, http://www.cfsan.fda.gov/∼ebam/bam-5.html, 2007). Comparable results were obtained by both methods. Only live Salmonella cells could be detected by this qRT-PCR assay, thus avoiding the dangers of false-positive results from nonviable cells. False negatives (inhibition of the PCR) were also ruled out through the use of an RNA internal control. This assay allows for the fast and accurate detection of viable Salmonella spp. in spinach, tomatoes, and in both jalapeno and serrano peppers. PMID:19376910
We have developed a sensitive method for the determination of Salmonella by integrating a streptavidinylated aptamer (SA-aptamer) as a signal amplification unit along with a modified asymmetric polymerase chain reaction (PCR) technique into the surface of an SPR sensor chip. The gold film of the sensor was first modified with a thiolated probe, and the target sequence and SA-aptamer were then induced to form a sandwich structure. If SA is added, the SA-aptamer forms a complex with SA which will amplify the signal. Under optimal conditions, this sensing scheme has a linear response in the 50 pM to 200 nM range, and the lower detection limit is 20 pM (for a synthetic target sequence). This strategy was successfully applied to the determination of Salmonella bacteria at levels as low as 60 CFU mL−1. This biosensor is sensitive, selective and highly stable. These features make this strategy a promising and powerful screening tool to detect pathogens in food, and in clinical and environmental samples. (author)
目的:研究喹诺酮对沙门菌毒力岛基因hilA和invA的mRNA表达水平的影响.方法:用多阶段诱导方式体外筛选耐喹诺酮沙门菌株,用荧光定量PCR方法测定喹诺酮敏感株和耐药株的hilA和invA基因的mRNA表达水平.结果:与喹诺酮敏感株相比,喹诺酮耐药株的hilA和invA的mRNA的表达水平显著降低(P<0.01).结论:喹诺酮可导致沙门菌毒力相关基因表达水平降低,这意味着喹诺酮耐药株毒力和致病性的降低.%Objective To investigate the effects of quinolones on the expressions of SPI-1 genes hilA and invA mRNA. Methods Quinolone-resistant Salmonella mutants were obtained from quinolone-susceptible strains by multiple-passage induction with increasing concentrations of ciprofloxacin in vitro. The expressions of hilA and invA genes mRNA were determined by real-time fluorescent quantitative RT-PCR in quinolone-susceptible and-resistant Salmonella strains. Results The expressions of hilA and invA gene mRNA were significantly decreased in quinolone-resistant strains (clinically acquired or drug-induced in vitro) than those in quinolone-susceptible strains (P < 0.01). Conclusion Suppressed expression of virulence-associated genes of quinolone-resistant salmonella strains suggests reduced virulence and pathogenicity of these bacteria.
Allgayer, M C; Lima-Rosa, C A V; Weimer, T A; Rodenbusch, C R; Pereira, R A; Streck, A F; Oliveira, S D; Canal, C W
Cloacal swabs were collected from 280 captive psittacine birds belonging to 13 species. Samples of dna were tested by PCR using a pair of primers that amplify a 284 base pair fragment of the Salmonella genus invA gene, and the PCR-positive samples were tested by standard microbiological techniques. Thirteen per cent of the samples were positive by PCR, but negative by microbiological techniques. The infection rates were significantly different among the 13 species, the most commonly infected being Amazona amazonica (28 per cent) and Amazona pretrei (20 per cent). Specific tests for Salmonella Typhimurium Salmonella Enteritidis, Salmonella Pullorum and Salmonella Gallinarum did not produce positive results. PMID:18567929
The cytotoxic and mutagenic specificity of two therapeutically employed psoralens was examined in several Ames Salmonella typhimurium strains with near ultraviolet light activation. Photomutagenic activity of 8-methoxypsoralen (8MOP) and 4,5',8-trimethylpsoralen (TMP) was found to be sequence-specific, and additionally was dependent on the level of DNA-repair proficiency. Phototoxicity was essentially identical in hisC3076, hisD3052 and hisG46 strains; uvrB- excision-repair-deficient bacteria were considerably more susceptible to lethal effects than wild-type parental strains. Finally, the data show that psoralens are potent frameshift photomutagens in Salmonella hisC3076 strains and demonstrate the potential utility of these strains in evaluating photomutagenic and phototoxic activity of new furocoumarin derivatives. (Auth.)
Full Text Available Polymerase chain reaction (PCR assay had been used to detect Salmonella in food and beverage samples using suitable primers which are based on specific invA gene of Salmonella. Twenty nine samples were collected from street food counters and some canteens in Margonda Street, Depok, West Java, Indonesia. It was found that five of twenty nine samples were detected to contain Salmonella and showed the presence of the amplified product of the size 244 bp. The method of PCR demonstrated the specificity of invA primers for detection of Salmonella as confirmed by biochemical and serological assay. The results of this study revealed that PCR was a rapid and useful tool for detection of Salmonella in food and beverage samples.
Full Text Available Shrimp is an important non-oil commodity for foreign trade in Indonesia. However, rejection of shrimp exports by the importing countries is still commonly encountered. In 2011, the USFDA recorded two cases of Salmonella spp. contamination in shrimp products from two shrimp processing companies in Indonesia. Analysis of Salmonella spp. in seafood is generally performed using a conventional method which takes at least 5 days. The objective of the study is to get a Salmonellae rapid detection method in shrimp by PCR. In this study, optimization of PCR protocol method to detect Salmonella invA gene was conducted using six different annealing temperatures (59, 59.5, 60.8, 62, 64 and 64.5°C. The results showed that 64°C was the optimum annealing temperature to detect the 284 bp fragment of Salmonella invA gene. The PCR based detection method has a DNA detection limit of 27.81ug/mL and 10°CFU/mL of viable salmonellae with 100% specificity. The PCR protocol is capable of detecting six different Salmonella serovars (S. Enteritidis, S. Hadar, S. Heidelberg, S. Kentucky, S. Paratyphi and S. Typhimurium but none of the non salmonellae isolates. Application of the PCR assay on Salmonella in shrimp after the selective enrichment step suggested that all 16 samples were positive for Salmonella. At the same time, the conventional method could only detected 3 samples for Salmonella positive.
Scientifically based and clinically validated new tools and methods to combat Salmonella infection in poultry, allowing to ensure the safety and health safety products - eggs and poultry meat. The method of selective decontamination involves the use of bivalent bacteriophage that is based on highly selected phages Phagum Salmonella typhimurium and Phagum Salmonella enteritidis, as well as probiotic laktobifadola. The developed tools and methods of selective decontamination followed by immuniz...
The standard method for detecting Salmonella generally analyzes food or fecal samples. Salmonella often occur in relatively low concentrations in environmental waters. Therefore, some form of concentration and proliferation may be needed. This study compares three Salmonella analysis methods and develops a new Salmonella detection procedure for use in environmental water samples. The new procedure for Salmonella detection include water concentration, nutrient broth enrichment, selection of Salmonella containing broth by PCR, isolation of Salmonella strains by selective culture plates, detection of possible Salmonella isolate by PCR, and biochemical testing. Serological assay and pulsed-field gel electrophoresis (PFGE) can be used to identify Salmonella serotype and genotype, respectively. This study analyzed 116 raw water samples taken from 18 water plants and belonging to 5 watersheds. Of these 116, 10 water samples (8.6%) taken from 7 water plants and belonging to 4 watersheds were positive for a Salmonella-specific polymerase chain reaction targeting the invA gene. Guided by serological assay results, this study identified 7 cultured Salmonella isolates as Salmonella enterica serovar: Alnaby, Enteritidis, Houten, Montevideo, Newport, Paratyphi B var. Java, and Victoria. These seven Salmonella serovars were identified in clinical cases for the same geographical areas, but only one of them was 100% homologous with clinical cases in the PFGE pattern. - Research highlights: → A new Salmonella detecting procedure for environmental water is developed. → Salmonella isolates are identified by serological assay and PFGE. → A total of seven Salmonella serovars is isolated from environmental water.
Fong, Karen; Wang, Siyun
In North America, outbreaks of Salmonella have been linked to low-water activity (aw) foods, such as nuts and seeds. These outbreaks have implicated an assortment of Salmonella serotypes. Some Salmonella serotypes (e.g., Enteritidis and Typhimurium) cause high proportions of salmonellosis. Nevertheless, there has recently been an emergence of uncommon Salmonella serotypes and strains (e.g., Tennessee, Hartford, and Thompson) in low-aw foods. The aim of this study was to evaluate the survival characteristics of Salmonella serotypes Enteritidis, Typhimurium, Tennessee, Hartford, and Thompson in three low-aw food ingredients with varying aw: peanut oil (aw = 0.521 ± 0.003), peanut shell (aw = 0.321 ± 0.20), and chia seeds (aw = 0.585 ± 0.003). The survival of individual Salmonella strains on each food matrix was monitored for a maximum of 150 days by spreading the bacterial cells onto Luria-Bertani and/or xylose lysine deoxycholate agar. Overall, Salmonella survived for the longest periods of time in peanut oil (96 ± 8 days), followed by chia seeds (94 ± 46 days). The survival period was substantially reduced on the surface of peanut shell (42 ± 49 h), although PCR after 70 days of incubation revealed the presence of Salmonella cells. In addition, Salmonella exhibited a strain-specific response in the three low-aw foods tested. Salmonella Hartford was identified as highly persistent in all low-aw food matrices, whereas Salmonella Typhimurium was the least persistent. The current research emphasizes the adaptable nature of Salmonella to low-aw food ingredients. This may pose additional problems owing to the downstream production of various end products. Additionally, unique survival characteristics among Salmonella strains highlight the need for tailored mitigation strategies regarding high-risk Salmonella strains in the food industry. PMID:26939645
... Linked to Small Turtles More Reports, Publications, and Communication Materials Surveillance Salmonella Atlas MMWR Articles References Communication ... Infections Linked to Red and Black Pepper/Italian-Style Meats 2006 through 2009 Outbreaks Saintpaul Infections Linked ...
Gonçalves, Flávio, 1929-1988
O presente trabalho, intitulado: “A Guerra Absoluta de Clausewitz nas Invasões Francesas”, tem como principal objetivo identificar os aspetos do conceito de Guerra Absoluta de Clausewitz que ocorreram durante o período das Invasões Francesas. Pretende-se esclarecer a importância do povo durante o período das Invasões Francesas, facto que deve ser de conhecimento público, de modo que exibam orgulho e tenham perfeita noção daquil...
Salmonella are facultative anaerobic Gram-negative non-spore forming rods belonging to the family Enterobacteriaceae. Salmonellosis is a zoonotic and foodborne illness that is usually transmitted by the fecal-oral route estimated to be responsible for 1.4 million cases of human infections in 2009 in...
Full Text Available Antimicrobial resistant Salmonella are among the leading causes of foodborne infections. Our aim was to determine Salmonella contamination during cattle slaughter in South African rural abattoirs (n = 23 and environmental samples. Furthermore, antimicrobial resistance patterns of the Salmonella isolates were determined. Samples of cattle faeces (n = 400, carcass sponges (n = 100, intestinal contents (n = 62, hides (n = 67, and water from the abattoirs (n = 75 were investigated for Salmonella species using microbiological techniques and species-specific polymerase chain reaction targeting the invA gene. In total 92 Salmonella species isolates were recovered. The Salmonella mean frequency of occurrence on hides, carcasses, and intestinal contents was 35.37% (n = 81. Eleven faecal samples (2.75% tested positive for Salmonella. The predominant serovar was Salmonella Enteritidis. Diverse serovars that were identified on carcasses were not necessarily found on the hides and intestinal contents. The inconsistent occurrence of the diverse Salmonella serovars on hides, carcasses, and intestinal contents implies that in addition to carriage on hides and in intestinal contents, other external factors also play an important role regarding carcass contamination. The 92 Salmonella were serotyped and tested for susceptibility towards the following antimicrobials: ampicillin, cefotaxime, enrofloxacin, kanamycin, and oxytetracycline using the disk diffusion method. Most Salmonella (n = 66; 71.7% isolates were resistant to at least one antimicrobial with highest resistance observed towards oxytetracycline (51.90%, which highlights the need for strict hygiene during slaughter and prudent antimicrobial use during animal production. In conclusion, cattle slaughtered in South African rural abattoirs harbour diverse Salmonella serovars that are resistant to antimicrobials, which could be a public health risk. The findings should assist policymakers with improving
Ktsoyan, Zhanna; Ghazaryan, Karine; Manukyan, Gayane; Martirosyan, Anush; Mnatsakanyan, Armine; Arakelova, Karine; Gevorgyan, Zaruhi; Sedrakyan, Anahit; Asoyan, Ara; Boyajyan, Anna; Aminov, Rustam
The main purpose of this study was to investigate the profile of inflammatory response in patients with acute salmonellosis caused by two serotypes of Salmonella enterica, S. Enteritidis and S. Typhimurium, as well as in convalescent patients with previous acute disease caused by S. Enteritidis. Patients with acute disease showed significantly elevated levels of IL-1β, IL-17, IL-10, and calprotectin compared to healthy control subjects. In convalescent patients, these markers were also significantly elevated, with the exception of IL-1β. Multivariate statistical analyses with the use of these variables produced models with a good predictive accuracy resulting in excellent separation of the diseased and healthy cohorts studied. Overall, the results suggest that the profile of inflammatory response in this disease is determined, to a significant degree, by the serotype of Salmonella, and the profile of certain cytokines and calprotectin remains abnormal for a number of months following the acute disease stage. PMID:26904722
Anna Boyajyan; Ara Asoyan; Anahit Sedrakyan; Zaruhi Gevorgyan; Karine Arakelova; Armine Mnatsakanyan; Anush Martirosyan; Gayane Manukyan; Karine Ghazaryan; Zhanna Ktsoyan; Rustam Aminov
The main purpose of this study was to investigate the profile of inflammatory response in patients with acute salmonellosis caused by two serotypes of Salmonella enterica, S. Enteritidis and S. Typhimurium, as well as in convalescent patients with previous acute disease caused by S. Enteritidis. Patients with acute disease showed significantly elevated levels of IL-1β, IL-17, IL-10, and calprotectin compared to healthy control subjects. In convalescent patients, these markers were also signif...
Full Text Available Background: Probiotic strains of bacteria can prevent Salmonella from causing disease by preventing the pathogen from colonizing the intestines. Two strains of probiotics, Lactobacillus acidophilius and Pediococcus spp, that were obtained from poultry fecal samples have been shown to be efficacious in poultry. The objective of this study was to determine if these strains of probiotics could prevent salmonellosis in a mouse model. Methods: First, both strains of probiotics were evaluated for in vitro efficacy to inhibit the growth of and interfere with virulence gene regulation in Salmonella enterica. For in vivo efficacy, mice was used which models Typhoid illness. Mice were divided into 2 groups: Control and treatment, Lactobacillus and Pediococcus (LP; 108 Log CFU. Two experiments were conducted. In the first experiment, the mice were treated with LP in water for the first two days of the experiment and challenged with Salmonella at day three. In the second experiment, the LP treatment was given in the water for 10 days and challenge was performed on day 11. In both experiments, at day 20 post-challenge, all mice were sacrificed, intestinal tracts and organs removed and cultured for Salmonella. Results: The probiotic strains inhibited the growth of Salmonella and down-regulation of virulence genes was noted, but dependent on the strain of Salmonella being evaluated. For the in vivo experiment, the probiotics did not afford the mice protection from infection and increasing the length of time the probiotics were administered did not improve the efficacy of the probiotics. Conclusions: It appears that these strains of probiotic bacteria are effective against Salmonella in vitro. However, these isolates did not afford protection from Salmonella infection to mice which may be due to host specifity as these isolates were obtained from poultry
Oueslati, Walid; Rjeibi, Mohamed Ridha; Mhadhbi, Moez; Jbeli, Mounir; Zrelli, Samia; Ettriqui, Abdelfettah
The aim of this work was to investigate the presence of Salmonella spp. in 300 beef meat samples collected from cattle carcasses of different categories (young bulls, culled heifers and culled cows). The detection of Salmonella spp. was performed by the alternative VIDAS Easy Salmonella technique and confirmed by PCR using Salmonella specific primers. Salmonella serotypes were determined by slide agglutination tests. The resistance to 12 antibiotics was determined by the diffusion method on Mueller-Hinton agar antibiotic discs. The overall contamination rate of beef by Salmonella spp. was 5.7% (17/300). This rate varied from naught (0/100) in bulls' meat to 14% (14/100) in culled cows' meat (pacid (6/17). All the strains were sensitive to the association (Amoxicillin+Clavulanic acid), Cefoxitin and Ceftazidime. In addition, our study showed that all Salmonella strains (17) were positive for invasion gene invA and negative for the virulence gene spvC. Only one isolate (S. Kentucky) harbored the h-li virulence gene. PMID:27183540
Lilic, Mirjana; Quezada, Cindy M.; Stebbins, C. Erec, E-mail: email@example.com [Laboratory of Structural Microbiology, The Rockefeller University, New York, NY 10065 (United States)
The cytoplasmic domain of Salmonella InvA shares homology to a recurring scaffold in the membrane-spanning components of the type II and type III secretion systems. Protein type III secretion systems (T3SSs) are organic nanosyringes that achieve an energy-dependent translocation of bacterial proteins through the two membranes of Gram-negative organisms. Examples include the pathogenic systems of animals, plants and symbiotic bacteria that inject factors into eukaryotic cells, and the flagellar export system that secretes flagellin. T3SSs possess a core of several membrane-associated proteins that are conserved across all known bacterial species that use this system. The Salmonella protein InvA is one of the most highly conserved proteins of this core of critical T3SS components. The crystal structure of a C-terminal domain of InvA reveals an unexpected homology to domains that have been repeatedly found as building blocks of other elements of the T3SS apparatus. This suggests the surprising hypothesis that evolution has produced a significant component of the apparatus structure through a series of gene-duplication and gene-rearrangement events.
The cytoplasmic domain of Salmonella InvA shares homology to a recurring scaffold in the membrane-spanning components of the type II and type III secretion systems. Protein type III secretion systems (T3SSs) are organic nanosyringes that achieve an energy-dependent translocation of bacterial proteins through the two membranes of Gram-negative organisms. Examples include the pathogenic systems of animals, plants and symbiotic bacteria that inject factors into eukaryotic cells, and the flagellar export system that secretes flagellin. T3SSs possess a core of several membrane-associated proteins that are conserved across all known bacterial species that use this system. The Salmonella protein InvA is one of the most highly conserved proteins of this core of critical T3SS components. The crystal structure of a C-terminal domain of InvA reveals an unexpected homology to domains that have been repeatedly found as building blocks of other elements of the T3SS apparatus. This suggests the surprising hypothesis that evolution has produced a significant component of the apparatus structure through a series of gene-duplication and gene-rearrangement events
Johnson, M. S.; Taylor, B L
Three methods of ATP depletion in Salmonella typhimurium were compared. ATP concentrations were lowest after arsenate treatment. Arsenate or alpha-methylglucoside-plus-azide treatment nonspecifically lowered all nucleotide triphosphate levels. Histidine starvation in a hisF mutant was relatively specific for ATP depletion and therefore has potential in distinguishing ATP-dependent processes from processes dependent on other nucleotides.
Swarmistha Devi Aribam
Full Text Available Salmonella-specific antibodies play an important role in host immunity; however, the mechanisms of Salmonella clearance by pathogen-specific antibodies remain to be completely elucidated since previous studies on antibody-mediated protection have yielded inconsistent results. These inconsistencies are at least partially attributable to the use of polyclonal antibodies against Salmonella antigens. Here, we developed a new monoclonal antibody (mAb-449 and identified its related immunogen that protected BALB/c mice from infection with Salmonella enterica serovar Typhimurium. In addition, these data indicate that the mAb-449 immunogen is likely a major protective antigen. Using in vitro infection studies, we also analyzed the mechanism by which mAb-449 conferred host protection. Notably, macrophages infected with mAb-449-treated S. Typhimurium showed enhanced pathogen uptake compared to counterparts infected with control IgG-treated bacteria. Moreover, these macrophages produced elevated levels of pro-inflammatory cytokine TNFα and nitric oxide, indicating that mAb-449 enhanced macrophage activation. Finally, the number of intracellular bacteria in mAb-449-activated macrophages decreased considerably, while the opposite was found in IgG-treated controls. Based on these findings, we suggest that, although S. Typhimurium has the potential to survive and replicate within macrophages, host production of a specific antibody can effectively mediate macrophage activation for clearance of intracellular bacteria.
Kuo, C.; Hsu, B.; Shen, T.; Tseng, S.; Tsai, J.; Huang, K.; Kao, P.; Chen, J.
Salmonella spp. is a common water-borne pathogens and its genus comprises more than 2,500 serotypes. Major pathogenic genotypes which cause typhoid fever, enteritis and other intestinal-type diseases are S. Typhimurium, S. Enteritidis, S. Stanley, S. Agona, S.Albany, S. Schwarzengrund, S. Newport, S. Choleraesuis, and S. Derby. Hence, the identification of the serotypes of Salmonella spp. is important. In the present study, the analytical procedures include direct concentration method, non-selective pre-enrichment method and selective enrichment method of Salmonella spp.. Both selective enrichment method and cultured bacteria were detected with specific primers of Salmonella spp. by polymerase chain reaction (PCR). At last, the serotypes of Salmonella were confirmed by using MLST (multilocus sequence typing) with aroC, dnaN, hemD, hisD, purE, sucA, thrA housekeeping genes to identify the strains of positive samples. This study contains 121 samples from three different types of water sources including the drinking water (51), streams (45), and swine wastewater (25). Thirteen samples with positive invA gene are separated from culture method. The strains of these positive samples which identified from MLST method are S. Albany, S. Typhimurium, S. Newport, S. Bareilly, and S. Derby. Some of the serotypes, S. Albany, S. Typhimurium and S. Newport, are highly pathogenic which correlated to human diarrhea. In our results, MLST is a useful method to identify the strains of Salmonella spp.. Keywords: Salmonella, PCR, MLST.
Malorny, B.; Hoorfar, Jeffrey; Bunge, C.;
Salmonella spp. and 47 non-Salmonella strains. The most selective primer set was found to be 139-141 (K. Rahn, S. A. De Grandis, R. C. Clarke, S. A. McEwen, J. E. Galan, C. Ginocchio, R. Curtiss 111, and C. L. Gyles, Mol. Cell. Probes 6:271-279, 1992), which targets the invA gene. An extended determination...
Rodriguez-Lazaro, D.; Hernández, Marta; Esteve, T.;
The aim of this work was the validation of a rapid, real-time PCR assay based on TaqMan((R)) technology for the unequivocal identification of Salmonella spp. to be used directly on an agar-grown colony. A real-time PCR system targeting at the Salmonella spp. invA gene was optimized and validated ...
Perelle, Sylvie; Dilasser, Françoise; Malorny, Burkhard; Grout, Joël; Hoorfar, Jeffrey; Fach, Patrick
corresponds respectively to 50 and 10 cells per PCR tube. Data on artificially contaminated samples indicated that both PCR methods were able to detect after enrichment less than five Salmonella cells in 25 g of food, giving 100% concordance with the ISO 6579 reference method. The results on naturally......In a previous study, we reported the performance of a PCR assay amplifying 285-bp of the invA gene of Salmonella spp. through an international ring-trial involving four participating laboratories [Int. J. Food Microbiol. 89 (2003) 241]. Based on the validated set of primers and recent advancements...... in PCR technology, we have designed two specific PCR assays for detecting Salmonella spp. We have compared PCR-enzyme-linked immunosorbent assay (PCR-ELISA) and LightCycler real-time PCR assay (LC-PCR) with the standard ISO 6579 bacteriological reference method. The two PCR tests incorporated an...
Full Text Available In this paper, the antibody titre to Salmonella enteritidis (SE was examined by the ELISA method in two flocks of laying hens, where during routine bacteriological investigations Salmonellae was never isolated, and in one flock where Colysepticemia was diagnosed and Salmonella isolated accidentally. In the flocks were Salmonellae were not isolated, a titre with a high level of specific antibodies to SE was discovered (15 and 45%, while the flock with accidental findings of SE was poorly positive (5%. These results point to the necessity of introducing serological monitoring to SE so that the infection of salmonella may be discovered early and the prevalence in the flock determined, and also for the purpose of applying adequate measures that could reduce the possibility of secretion of SE through eggs.
Sandra Patricia Rivera
Full Text Available Introduction: Chlorination is the most widely used disinfection process for drinking water production. The formation of chlorination carcinogenic by-products and chlorine intoxication by direct manipulation in small communities has motivated the study of alternative disinfection processes. In this sense, processes of advanced oxidation (PAOs have yielded promising results. Escherichia coli (E. coli is customarily used as faecal bacterial indicator to determine the efficiency of disinfection processes. However, it has been shown that E. coli is less resistant to disinfection than other enteric bacteria such as Shigella spp. and Salmonella spp. Additionally, the viable non-culturable (VNC state yields bacteria which are not detectable on many culture media. Objective: The main objective is to standardize a method for counting Salmonella spp. and Shigella spp. in specific liquid media to reliably quantify the bacteriological potential risk related to disinfection processes based on PAO. Methods: The study followed a randomized bi-factorial experimental design and the Duncan multiple comparison test. This design allowed the selection of specific liquid media to fittingly standardize the counting of Salmonella spp. and Shigella spp. Results: We found that the best broth for counting Salmonella typhimurium strain at different concentrations in pure and mixed cultures was the Rappaport broth RP, the EE broth also allowed growing the two bacterial species tested in this research. Nonetheless, the latter results suggest the use of additional tests for this particular broth. Discussion: There was a variation in the counting results when pure cultures were used compared to those obtained from mixtures of microorganisms. It was also noted that Salmonella typhimurium and Shigella sonnei, were recovered from minimal concentrations in both RP and EE broths, respectively. To some extent, this suggests an additional confirmative method when using the
Sandra Patricia Rivera
Full Text Available Introduction: Chlorination is the most widely used disinfection process for drinking water production. The formation of chlorination carcinogenic by-products and chlorine intoxication by direct manipulation in small communities has motivated the study of alternative disinfection processes. In this sense, processes of advanced oxidation (PAOs have yielded promising results. Escherichia coli (E. coli is customarily used as faecal bacterial indicator to determine the efficiency of disinfection processes. However, it has been shown that E. coli is less resistant to disinfection than other enteric bacteria such as Shigella spp. and Salmonella spp. Additionally, the viable non-culturable (VNC state yields bacteria which are not detectable on many culture media.Objective: The main objective is to standardize a method for counting Salmonella spp. and Shigella spp. in specific liquid media to reliably quantify the bacteriological potential risk related to disinfection processes based on PAO.Methods: The study followed a randomized bi-factorial experimental design and the Duncan multiple comparison test. This design allowed the selection of specific liquid media to fittingly standardize the counting of Salmonella spp. and Shigella spp.Results: We found that the best broth for counting Salmonella typhimurium strain at different concentrations in pure and mixed cultures was the Rappaport broth RP, the EE broth also allowed growing the two bacterial species tested in this research. Nonetheless, the latter results suggest the use of additional tests for this particular broth.Discussion: There was a variation in the counting results when pure cultures were used compared to those obtained from mixtures of microorganisms. It was also noted that Salmonella typhimurium and Shigella sonnei, were recovered from minimal concentrations in both RP and EE broths, respectively. To some extent, this suggests an additional confirmative method when using the EE® broth
Full Text Available Salmonella enterica and Shigella species are commonly associated with food and water borne infections leading to gastrointestinal diseases. The present work was undertaken to develop a sensitive and reliable PCR based detection system for simultaneous detection of Salmonella enterica and Shigella at species level. For this the conserved regions of specific genes namely ipaH1, ipaH, wbgZ, wzy and invA were targeted for detection of Shigella genus, S. flexneri, S. sonnei, S. boydii and Salmonella enterica respectively along with an internal amplification control (IAC. The results showed that twenty Salmonella and eleven Shigella spp., were accurately identified by the assay without showing non-specificity against closely related other Enterobacteriaceae organisms and also against other pathogens. Further evaluation of multiplex PCR was undertaken on 50 natural samples of chicken, eggs and poultry litter and results compared with conventional culture isolation and identification procedure. The multiplex PCR identified the presence of Salmonella and Shigella strains with a short pre-enrichment step of 5 h in peptone water and the same samples were processed by conventional procedures for comparison. Therefore, this reported multiplex PCR can serve as an alternative to the tedious time-consuming procedure of culture and identification in food safety laboratories.
Analysis of Salmonella enterica Serotype-Host Specificity in Calves: Avirulence of S. enterica Serotype Gallinarum Correlates with Bacterial Dissemination from Mesenteric Lymph Nodes and Persistence In Vivo
Paulin, Susan M.; Watson, Patricia R.; Benmore, Annette R.; Stevens, Mark P.; Jones, Philip W.; Villarreal-Ramos, Bernardo; Wallis, Timothy S.
Host and bacterial factors that determine whether Salmonella serotypes remain restricted to the gastrointestinal tract or penetrate beyond the mucosa and cause systemic disease remain largely undefined. Here, factors influencing Salmonella host specificity in calves were assessed by characterizing the pathogenesis of different serotypes. Salmonella enterica serotype Dublin was highly virulent intravenously, whereas S. enterica serotype Choleraesuis was moderately virulent. Both serotypes were...
Amruta Nair; T. Balasaravanan; MALIK, S V S; Vysakh Mohan; Manesh Kumar; Jess Vergis; Rawool, Deepak B.
Aim: This study was carried out to determine the prevalence, distribution, and identification of Salmonella serotypes in diarrheagenic infants and young animals, including sewage waste and fresh vegetables. Materials and Methods: A total of 550 samples were processed for the isolation of Salmonella spp., using standard microbiological and biochemical tests. Further polymerase chain reaction (PCR) detection of Salmonella genus was carried out using self-designed primers targeting invA gene ...
Salmonella are the main responsible agent for the frequent food borne gastrointestinal diseases. In Tunisia, this pathogen is considered one of the most important causes of toxiinfections and its detection using classical methods is laborious and requires a large amount of time for revelation. To solve this problem, we developed a rapid molecular technique for the detection of the invA virulence gene sequence which is found in the majority of Salmonella spp. This technique is a hemi nested PCR amplification using specific primers designed and by bioinformatics tools. The detection method consisted of pre-enrichment of the sample in buffered peptone water (BPW), followed by a total DNA extraction step prior to single tube hemi nested PCR amplification. This method was found highly specific and sensitive to detect low levels of salmonella typhimurium and salmonella enteritidis (1cfu/ 25g) in naturally contaminated spicy sausage (merguez) samples. These results can benefit the public health agencies concerning microbiological and quality aspects of the commercial and traditional merguez meat production in Tunisia. (Author)
Malorny, B.; Hoorfar, Jeffrey; Hugas, M.;
A collaborative study involving four European laboratories was conducted to investigate the diagnostic accuracy of a Salmonella specific PCR-based method, which was evaluated within the European FOOD-PCR project (http://www.pcr.dk). Each laboratory analysed by the PCR a set of independent obtained...... presumably naturally contaminated samples and compared the results with the microbiological culture method. The PCR-based method comprised a preenrichment step in buffered peptone water followed by a thermal cell lysis using a closed tube resin-based method. Artificially contaminated minced beef and whole...... investigated. The interlaboratory diagnostic accuracy, i.e. diagnostic specificity and sensitivity, was shown to be 97.5%. The co-amplification of an internal amplification control indicated possible inhibitory substances derived from the sample. This work can contribute to the quality assurance of PCR...
Soyer, Yeşim; Orsi, Renato H.; Rodriguez-Rivera, Lorraine D; Sun, Qi; Wiedmann, Martin
Background The bacterium Salmonella enterica includes a diversity of serotypes that cause disease in humans and different animal species. Some Salmonella serotypes show a broad host range, some are host restricted and exclusively associated with one particular host, and some are associated with one particular host species, but able to cause disease in other host species and are thus considered "host adapted". Five Salmonella genome sequences, representing a broad host range serotype (Typhimur...
Kim, Jeongjin; Jo, Ara; Ding, Tian; Lee, Hyeon-Yong; Ahn, Juhee
This study describes a new effort toward understanding the interaction mechanisms between antibiotic-resistant Salmonella Typhimurium and phages. The antibiotic susceptibility, β-lactamase activity, bacterial motility, gene expression, and lytic activity were evaluated in ciprofloxacin-induced antibiotic-sensitive Salmonella Typhimurium (ASST(CIP)) and ciprofloxacin-induced antibiotic-resistant S. Typhimurium (ARST(CIP)), which were compared to the wild-type strains (ASST(WT) and ARST(WT)). The MIC values of ampicillin, norfloxacin, chloramphenicol, and tetracycline were significantly increased to > 512, 16, 16, and 256 μg/ml, respectively, in the ARST(CIP). The lowest and highest extracellular lactamase activities were observed in ASST(WT) (6.85 μmol/min/ml) and ARST(CIP) (48.83 μmol/min/ml), respectively. The acrA, lpfE, and hilA genes were significantly upregulated by more than tenfold in both ASST(CIP) and ARST(CIP). The induction of multiple antibiotic resistance resulted from the increased efflux pump activity (AcrAB-TolC). The highest phage adsorption rates were more than 95 % for ASST(WT), ASST(CIP), and ARST(WT), while the lowest adsorption rate was 52 % for ARST(CIP) at 15 min of infection. The least lytic activity of phage was 20 % against the ARST(CIP), followed by ASST(CIP) (30 %). The adsorption rate of phage against ARST(CIP) was 52 % at 15 min of infection, which resulted in the decrease in lytic activity (12 %). Understanding the interaction of phage and bacteria is essential for the practical application of phage to control and detect antibiotic-resistant bacteria. The results provide useful information for understanding the binding specificity of phages for multiple antibiotic-resistant pathogens. PMID:27000396
Salmonella is one of the primary foodborne pathogens associated with swine production and represents a significant threat to human health. Bacteriophage are naturally-occurring viruses that prey on bacteria and have been suggested as a potential intervention strategy to reduce Salmonella in food an...
Kohler, Amanda C. [Rockefeller University, New York, NY 10065 (United States); Spanò, Stefania; Galán, Jorge E. [Yale University School of Medicine, New Haven, CT 06536 (United States); Stebbins, C. Erec, E-mail: firstname.lastname@example.org [Rockefeller University, New York, NY 10065 (United States)
The Salmonella effector protein GtgE functions as a cysteine protease to cleave a subset of the Rab-family GTPases and to prevent delivery of antimicrobial agents to the Salmonella-containing vacuole. GtgE is an effector protein from Salmonella Typhimurium that modulates trafficking of the Salmonella-containing vacuole. It exerts its function by cleaving the Rab-family GTPases Rab29, Rab32 and Rab38, thereby preventing the delivery of antimicrobial factors to the bacteria-containing vacuole. Here, the crystal structure of GtgE at 1.65 Å resolution is presented, and structure-based mutagenesis and in vivo infection assays are used to identify its catalytic triad. A panel of cysteine protease inhibitors were examined and it was determined that N-ethylmaleimide, antipain and chymostatin inhibit GtgE activity in vitro. These findings provide the basis for the development of novel therapeutic strategies to combat Salmonella infections.
The Salmonella effector protein GtgE functions as a cysteine protease to cleave a subset of the Rab-family GTPases and to prevent delivery of antimicrobial agents to the Salmonella-containing vacuole. GtgE is an effector protein from Salmonella Typhimurium that modulates trafficking of the Salmonella-containing vacuole. It exerts its function by cleaving the Rab-family GTPases Rab29, Rab32 and Rab38, thereby preventing the delivery of antimicrobial factors to the bacteria-containing vacuole. Here, the crystal structure of GtgE at 1.65 Å resolution is presented, and structure-based mutagenesis and in vivo infection assays are used to identify its catalytic triad. A panel of cysteine protease inhibitors were examined and it was determined that N-ethylmaleimide, antipain and chymostatin inhibit GtgE activity in vitro. These findings provide the basis for the development of novel therapeutic strategies to combat Salmonella infections
Lilic, M.; Quezada, C; Stebbins, C
Protein type III secretion systems (T3SSs) are organic nanosyringes that achieve an energy-dependent translocation of bacterial proteins through the two membranes of Gram-negative organisms. Examples include the pathogenic systems of animals, plants and symbiotic bacteria that inject factors into eukaryotic cells, and the flagellar export system that secretes flagellin. T3SSs possess a core of several membrane-associated proteins that are conserved across all known bacterial species that use this system. The Salmonella protein InvA is one of the most highly conserved proteins of this core of critical T3SS components. The crystal structure of a C-terminal domain of InvA reveals an unexpected homology to domains that have been repeatedly found as building blocks of other elements of the T3SS apparatus. This suggests the surprising hypothesis that evolution has produced a significant component of the apparatus structure through a series of gene-duplication and gene-rearrangement events.
Fong, Karen; Wang, Siyun
Cross-protection represents a considerable challenge in the food industry where hurdled interventions are often employed to reduce Salmonella contamination. The heat resistance of Salmonella strains from five serotypes (i.e., Typhimurium, Enteritidis, Tennessee, Thompson and Hartford) at 70 °C was determined by measurement of viable cell populations before and after adaptation to two common stresses employed in low-water activity food processing, desiccation and sub-lethal heat treatment. Survival of Salmonella at 70 °C significantly increased (p day incubation in peanut oil (aw 0.52 ± 0.00) and/or the exposure to a sub-lethal heat treatment at 45 °C for 3 min. Quantitative PCR revealed upregulation of two desiccation stress-related genes, fadA and otsB, following the peanut oil incubation, whereas heat treatment induced upregulation of a heat-resistance gene, dnaK. Invasion gene invA and alternative sigma factor rpoE were downregulated following either of the treatments. Interestingly, different Salmonella strains yielded different transcriptional profiles. The strain-specific resistance phenotypes and transcriptional profiles provided further insights into the mechanisms employed to tolerate desiccation and heat stresses in the food industry. PMID:27217370
Abstract Pork meat is one of the major sources for human infections with Salmonella enterica subspecies enterica serovars. Further, zoonoses caused by Salmonella 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 met...
Van der Gaag, M.A.; Enting, J.; Mul, Monique F.
In The Netherlands, a system to control the prevalence of Salmonella in the pork supply chain is under development by the Product Boards of Livestock, Meat and Eggs. Therefore, the Animal Sciences Group-Applied Research has developed a tool to support pig-finishing farms to control Salmonella. This tool contains eight checklists. The checklists are based on the principles of the HACCP methodology. Important hazards and control measures were determined. Each checklist deals with a part of the ...
Full Text Available Real-time polymerase chain reaction (PCR allows rapid detection of Salmonella in frozen dairy products, but it might cause a false positive detection result because it might amplify DNA from dead target cells as well. In this study, Salmonella-free frozen ice cream was initially inoculated with heat-killed Salmonella Typhimurium cells and stored at −18°C. Bacterial DNA extracted from the sample was amplified using TaqMan probe-based real-time PCR targeting the invA gene. Our results indicated that DNA from the dead cells remained stable in frozen ice cream for at least 20 days, and could produce fluorescence signal for real-time PCR as well. To overcome this limitation, propidium monoazide (PMA was combined with real-time PCR. PMA treatment can effectively prevent PCR amplification from heat-killed Salmonella cells in frozen ice cream. The PMA real-time PCR assay can selectively detect viable Salmonella at as low as 103 CFU/mL. Combining 18 hours of pre-enrichment with the assay allows for the detection of viable Salmonella at 100 CFU/mL and avoiding the false-positive result of dead cells. The PMA real-time PCR assay provides an alternative specifically for detection of viable Salmonella in ice cream. However, when the PMA real-time PCR assay was evaluated in ice cream subjected to frozen storage, it obviously underestimated the contamination situation of viable Salmonella, which might lead to a false negative result. According to this result, the use of enrichment prior to PMA real-time PCR analysis remains as the more appropriate approach.
Analysis of Salmonella enterica serotype-host specificity in calves: Avirulence of S-enterica serotype gallinarum correlates with bacterial dissemination from mesenteric lymph nodes and persistence in vivo
Paulin, S M; Watson, P. R.; Benmore, A R; M.P. Stevens; Jones, P W; Villarreal-Ramos, B.; Wallis, T S
Host and bacterial factors that determine whether Salmonella serotypes remain restricted to the gastrointestinal tract or penetrate beyond the mucosa and cause systemic disease remain largely undefined. Here, factors influencing Salmonella host specificity in calves were assessed by characterizing the pathogenesis of different serotypes. Salmonella enterica serotype Dublin was highly virulent intravenously, whereas S. enterica serotype Choleraesuis was moderately virulent. Both serotypes were...
Monadi, M. (MSc
Full Text Available Background and Objective: Salmonellosis is the most common type of food poisoning in developed and developing countries that is caused by Salmonella serotype. Hence, we aimed to identify the Salmonella serovars in eggs obtained from Kohgiluyeh and Boyerahmad province and to evaluate antibiotic resistance of the isolated strains. Material and Methods: In this study, 210 eggs were collected from Kohgiluyeh and Boyerahmad Province. The bacteria were isolated and identified using biochemical tests. After extraction of genomic DNA, Salmonella gender, Salmonella enteritidis and Salmonella typhimurium were investigated by invA, fliC and sefA primers, respectively, using Multiplex PCR method. Results: Of 210, 14 (6.66% were contaminated with Salmonella. Of these, 12 (5.71% were Salmonella typhimurium and 2 (0.95% were related to Salmonella spp. None of the samples were contaminated with Salmonella enteritidis. The highest resistance was related to penicillin (100% and neomycin (78.57%. Conclusion: Salmonella typhimurium is the predominant serovar causing contamination in the eggs of this Province. Given the wide spread of antibiotic resistance in different serotypes of Salmonella, we recommend avoiding of indiscriminate use of antibiotics in livestock and poultry
Salmonella is among the most commonly known bacterial pathogens to cause human illness. Often Salmonellosis is associated with the consumption of contaminated foods like meat, eggs or egg products. However, during the last decades an increase of outbreaks is recognized to be caused by human pathogen
Pasetti, Marcela F; Salerno-Gonçalves, Rosangela; Sztein, Marcelo B
We investigated the ability of live attenuated Salmonella enterica serovar Typhi strains delivered to mice intranasally to induce specific cytotoxic T-lymphocyte (CTL) responses at regional and systemic levels. Mice immunized with two doses (28 days apart) of Salmonella serovar Typhi strain Ty21a, the licensed oral typhoid vaccine, and genetically attenuated mutants CVD 908 (DeltaaroC DeltaaroD), CVD 915 (DeltaguaBA), and CVD 908-htrA (DeltaaroC DeltaaroD DeltahtrA) induced CTL specific for Salmonella serovar Typhi-infected cells in spleens and cervical lymph nodes. CTL were detected in effector T cells that had been expanded in vitro for 7 days in the presence of Salmonella-infected syngeneic splenocytes. A second round of stimulation further enhanced the levels of specific cytotoxicity. CTL activity was observed in sorted alphabeta+ CD8+ T cells, which were remarkably increased after expansion, but not in CD4+ T cells. CTL from both cervical lymph nodes and spleens failed to recognize Salmonella-infected major histocompatibility complex (MHC)-mismatched cells, indicating that the responses were MHC restricted. Studies in which MHC blocking antibodies were used showed that H-2L(d) was the restriction element. This is the first demonstration that Salmonella serovar Typhi vaccines delivered intranasally elicit CD8+ MHC class I-restricted CTL. The results further support the usefulness of the murine intranasal model for evaluating the immunogenicity of typhoid vaccine candidates at the preclinical level. PMID:12117906
Salmonella is among the most commonly known bacterial pathogens to cause human illness. Often Salmonellosis is associated with the consumption of contaminated foods like meat, eggs or egg products. However, during the last decades an increase of outbreaks is recognized to be caused by human pathogenic bacteria in association with fresh produce. The use of manure for production of vegetables, e.g. lettuce, contributes significantly to the risk of contamination of fresh produce. Enteric pathoge...
Sílvia Dias de Oliveira; Carla Rosane Rodenbusch; Geovana B. Michael; Marisa I.R. Cardoso; Cláudio Wageck Canal; Adriano Brandelli
The presence of three virulence genes, invA, spvR, and spvC, was determined in Salmonella Enteritidis isolated from poultry, pigs, humans and food. All isolates were positive for the invA gene, with 91.2% being positive for spvR and 90.2% for spvC. There was no significant difference in the prevalence of the virulence genes between isolates from different sources. The results indicate that there is a putative high virulence potential for the S. Enteritidis isolates characterized.A presença de...
Robbe-Saule, Véronique; Lopes, Miguel Dias; Kolb, Annie; Norel, Françoise
The small regulatory protein Crl activates sigma(S) (RpoS), the stationary-phase and general stress response sigma factor. Crl has been reported to bind sigma(S) in vitro and to facilitate the formation of RNA polymerase holoenzyme. In Salmonella enterica serovar Typhimurium, Crl is required for the development of the rdar morphotype and transcription initiation of the sigma(S)-dependent genes csgD and adrA, involved in curli and cellulose production. Here, we examined the expression of other sigma(S)-dependent phenotypes and genes in a Deltacrl mutant of Salmonella. Gene fusion analyses and in vitro transcription assays indicate that the magnitude of Crl activation differs between promoters and is highly dependent on sigma(S) levels. We replaced the wild-type rpoS allele in S. enterica serovar Typhimurium strain ATCC 14028 with the rpoS(LT2) allele that shows reduced expression of sigma(S); the result was an increased Crl activation ratio and larger physiological effects of Crl on oxidative, thermal, and acid stress resistance levels during stationary phase. We also found that crl, rpoS, and crl rpoS strains grew better on succinate than did the wild type and expressed the succinate dehydrogenase sdhCDBA operon more strongly. The crl and rpoS(LT2) mutations also increased the competitive fitness of Salmonella in stationary phase. These results show that Crl contributes to negative regulation by sigma(S), a finding consistent with a role for Crl in sigma factor competition via the facilitation of sigma(S) binding to core RNA polymerase. PMID:17293430
Timmons, Chris; Dobhal, Shefali; Fletcher, Jacqueline; Ma, Li Maria
Foodborne illnesses caused by Salmonella enterica and Escherichia coli O157:H7 are worldwide health concerns. Rapid, sensitive, and robust detection of these pathogens in foods and in clinical and environmental samples is essential for routine food quality testing, effective surveillance, and outbreak investigations. The aim of this study was to evaluate the effect on PCR sensitivity of adding a short, AT-rich overhanging nucleotide sequence (flap) to the 5' end of PCR primers specific for the detection of Salmonella and E. coli O157:H7. Primers targeting the invA gene of Salmonella and the rfbE gene of E. coli O157:H7 were synthesized with or without a 12-bp, AT-rich 5' flap (5'-AATAAATCATAA-3'). Singleplex PCR, multiplex PCR, and real-time PCR sensitivity assays were conducted using purified bacterial genomic DNA and crude cell lysates of bacterial cells. The effect of background flora on detection was evaluated by spiking tomato and jalapeno pepper surface washes with E. coli O157:H7 and Salmonella Saintpaul. When targeting individual pathogens, end-point PCR assays using flap-amended primers were more efficient than nonamended primers, with 20.4 and 23.5% increases in amplicon yield for Salmonella and E. coli O157:H7, respectively. In multiplex PCR assays, a 10- to 100-fold increase in detection sensitivity was observed when the primer flap sequence was incorporated. This improvement in both singleplex and multiplex PCR efficiency and sensitivity can lead to improved Salmonella and E. coli O157:H7 detection. PMID:23575131
Full Text Available BACKGROUND: Serotype-specific PCR assays targeting Salmonella enterica serotypes Typhi and Paratyphi A, the causal agents of typhoid and paratyphoid fevers, are required to accelerate formal diagnosis and to overcome the lack of typing sera and, in some situations, the need for culture. However, the sensitivity and specificity of such assays must be demonstrated on large collections of strains representative of the targeted serotypes and all other bacterial populations producing similar clinical symptoms. METHODOLOGY: Using a new family of repeated DNA sequences, CRISPR (clustered regularly interspaced short palindromic repeats, as a serotype-specific target, we developed a conventional multiplex PCR assay for the detection and differentiation of serotypes Typhi and Paratyphi A from cultured isolates. We also developed EvaGreen-based real-time singleplex PCR assays with the same two sets of primers. PRINCIPAL FINDINGS: We achieved 100% sensitivity and specificity for each protocol after validation of the assays on 188 serotype Typhi and 74 serotype Paratyphi A strains from diverse genetic groups, geographic origins and time periods and on 70 strains of bacteria frequently encountered in bloodstream infections, including 29 other Salmonella serotypes and 42 strains from 38 other bacterial species. CONCLUSIONS: The performance and convenience of our serotype-specific PCR assays should facilitate the rapid and accurate identification of these two major serotypes in a large range of clinical and public health laboratories with access to PCR technology. These assays were developed for use with DNA from cultured isolates, but with modifications to the assay, the CRISPR targets could be used in the development of assays for use with clinical and other samples.
Full Text Available BACKGROUND: The phagocytic oxidative burst is a primary effector of innate immunity that protects against bacterial infection. However, the mechanism by which reactive oxygen species (ROS kill or inhibit bacteria is not known. It is often assumed that DNA is a primary target of oxidative damage, consistent with known effects of endogenously produced ROS in the bacterial cytoplasm. But most studies fail to distinguish between effects of host derived ROS versus damage caused by endogenous bacterial sources. We took advantage of both the ability of Salmonella enterica serovar Typhimurium to survive in macrophages and the genetic tractability of the system to test the hypothesis that phagocytic superoxide damages cytoplasmic targets including DNA. METHODOLOGY/PRINCIPAL FINDINGS: SodCI is a periplasmic Cu-Zn superoxide dismutase (SOD that contributes to the survival of Salmonella Typhimurium in macrophages. Through competitive virulence assays, we asked if sodCI has a genetic interaction with various cytoplasmic systems. We found that SodCI acts independently of cytoplasmic SODs, SodA and SodB. In addition, SodCI acts independently of the base excision repair system and RuvAB, involved in DNA repair. Although sodCI did show genetic interaction with recA, this was apparently independent of recombination and is presumably due to the pleiotropic effects of a recA mutation. CONCLUSIONS/SIGNIFICANCE: Taken together, these results suggest that bacterial inhibition by phagocytic superoxide is primarily the result of damage to an extracytoplasmic target.
Cossi, Marcus Vinícius Coutinho; Burin, Raquel Cristina Konrad; Lopes, Danilo Augusto; Dias, Mariane Rezende; Castilho, Natalia Parma Augusto de; de Arruda Pinto, Paulo Sérgiode; Nero, Luís Augusto
Salmonella can contaminate finished products of butcher shops, mainly through cross-contamination of utensils exposed to raw materials. To identify the main sources of contamination with this foodborne pathogen in four butcher shop environments, surface samples were obtained from employees' hands, cutting boards, knives, floor of the refrigeration room, meat grinders, and meat tenderizers (32 samples per area) and analyzed for Salmonella using the International Organization for Standardization method 6579, with modifications. Suspect isolates were identified by PCR (targeting ompC), and confirmed Salmonella isolates were subjected to pulsed-field gel electrophoresis (after treatment with restriction enzyme XbaI), analyzed for the presence of virulence genes (invA, sefA, and spvC), and screened for resistance to 12 antimicrobials. Salmonella isolates was identified only on cutting boards (five samples) from three butcher shops. Fifteen isolates were confirmed as Salmonella belonging to four pulse types (similarity of 71.1 to 100%). The invA gene was detected in 13 isolates, and the sefA was found in 8 isolates; no isolate carried spvC. All tested isolates were resistant to clindamycin and sensitive to amikacin and cefotaxine, and all isolates were resistant to at least 3 of the 12 antimicrobials tested. The results indicate the importance of cutting boards as a source of Salmonella contamination in butcher shops. The presence of multidrug-resistant Salmonella strains possessing virulence genes highlights the health risks for consumers. PMID:23992511
Michael Keusgen; Peter KÃƒÂ¤mpfer; Olga Lezrich; Saikat Datta Mazumdar; Benjamin Barlen
This study explores the possibility of simultaneous and specific detection ofSalmonella serovars by surface plasmon resonance (SPR). The PlasmonicÃ‚Â® SPR device wasused to develop this rapid assay. The sandwich immunoassay involves the use of apolyclonal anti-Salmonella antibody to simultaneous capture multiple Salmonella serovarspresent in a sample. This is followed by specific detection of the captured serovars usingO-specific anti-Salmonella antibodies. Milk spiked with Salmonella Typhimu...
... Salmonella Reading and Salmonella Abony Infections linked to Alfalfa Sprouts Advice to Consumers & Retailers Case Count Maps ... Signs & Symptoms Key Resources Salmonella Infections Linked to Alfalfa Sprouts from One Contaminated Seed Lot Advice to ...
Mashooq, Mohmad; Kumar, Deepak; Niranjan, Ankush Kiran; Agarwal, Rajesh Kumar; Rathore, Rajesh
A one step, single tube, accelerated probe based real time loop mediated isothermal amplification (RT LAMP) assay was developed for detecting the invasion gene (InvA) of Salmonella. The probe based RT LAMP is a novel method of gene amplification that amplifies nucleic acid with high specificity and rapidity under isothermal conditions with a set of six primers. The whole procedure is very simple and rapid, and amplification can be obtained in 20min. Detection of gene amplification was accomplished by amplification curve, turbidity and addition of DNA binding dye at the end of the reaction results in colour difference and can be visualized under normal day light and in UV. The sensitivity of developed assay was found 10 fold higher than taqman based qPCR. The specificity of the RT LAMP assay was validated by the absence of any cross reaction with other members of enterobacteriaceae family and other gram negative bacteria. These results indicate that the probe based RT LAMP assay is extremely rapid, cost effective, highly specific and sensitivity and has potential usefulness for rapid Salmonella surveillance. PMID:27130353
Jacobsen, Carsten Suhr; Holben, William E
Direct quantification of mRNA from Salmonella sp. seeded for 1 h to soil and chicken manure was accomplished using magnetic capture hybridization as a purification technique. This detection strategy targeted the invA gene present in Salmonella sp. After cell lysis, phenol/chloroform purification and isopropanol precipitation, the RNA extract was combined with the hybridization probe conjugated to paramagnetic beads. After hybridization, the captured nucleic acids were released by denaturation and purified of contaminating DNA using DNase. The resulting RNA was of high purity and there was no need for dilution of the samples prior to RT-PCR. The developed procedure was reproducibly used to quantify Salmonella sp. in high organic agricultural soil. The detection limit for mRNA using ordinary quantitative PCR (employing SYBRgreen-based detection) was 5 x 10(4)Salmonella sp. cells per gram of soil. Chicken manure amended into soil (1:4 w/w) did not reduce the ability to quantify Salmonella sp. mRNA in soil. Pasteurization (65 degrees C, 30 min) of chicken manure containing Salmonella sp. dramatically reduced the detection of invA mRNA (requiring 42 qPCR cycles for detection versus 26 cycles in unpasteurized manure), presumably due to degradation of the invA mRNA in Salmonella sp. cells killed by pasteurization. By contrast, DNA-based qPCR still detected Salmonella sp. in the pasteurized manure. Thus, in this case using samples seeded with fresh Salmonella sp. the mRNA-based detection appears to be superior to minimizing false-positive detection which was prevalent with DNA-based qPCR. PMID:17383760
Sun Qi; Rodriguez-Rivera Lorraine D; Orsi Renato H; Soyer Yeşim; Wiedmann Martin
Abstract Background The bacterium Salmonella enterica includes a diversity of serotypes that cause disease in humans and different animal species. Some Salmonella serotypes show a broad host range, some are host restricted and exclusively associated with one particular host, and some are associated with one particular host species, but able to cause disease in other host species and are thus considered "host adapted". Five Salmonella genome sequences, representing a broad host range serotype ...
Full Text Available Abstract Background The induction of antigen-specific CD8+ T cell cytokine responses against an attenuated, oral recombinant Salmonella enterica serovar Typhimurium vaccine expressing a green fluorescent protein (GFP model antigen was investigated. A GFP expression plasmid was constructed in which the gfp gene was fused in-frame with the 5' domain of the Escherichia coli β-galactosidase α-gene fragment with expression under the lac promoter. Groups of mice were orally immunized three times with the bacteria and systemic CD8+ T cell cytokine responses were evaluated. Results High level of the GFP model antigen was expressed by the recombinant Salmonella vaccine vector. Systemic GFP-specific CD8+ T cell cytokine (IFN-γ and IL-4 immune responses were detected after mice were orally vaccinated with the bacteria. It was shown that 226 net IFN-γ and 132 net IL-4 GFP-specific SFUs/10e6 splenocytes were formed in an ELISPOT assay. The level of IFN-γ produced by GFP peptide-stimulated cells was 65.2-fold above background (p Conclusion These results suggested that a high expressing recombinant Salmonella vaccine given orally to mice would elicit antigen-specific CD8+ T cell responses in the spleen. Salmonella bacteria may, therefore, be used as potential mucosal vaccine vectors.
Rosenshine, I.; Ruschkowski, S; Foubister, V; Finlay, B B
Salmonella typhimurium invades nonphagocytic epithelial and fibroblast cells via a process resembling phagocytosis. We have compared some phenotypes that are involved in S. typhimurium invasion by using different host cell lines, including HeLa, Henle-407, and A431. Infection with either wild-type S. typhimurium, bacterial culture supernatant, or the noninvasive invA mutant was associated with induction of tyrosine phosphorylation of host cell mitogenic activating protein kinase. However, we ...
Full Text Available Abstract Background Whole genome sequencing of bacteriophages suitable for biocontrol of pathogens in food products is a pre-requisite to any phage-based intervention procedure. Trials involving the biosanitization of Salmonella Typhimurium in the pig production environment identified one such candidate, ΦSH19. Results This phage was sequenced and analysis of its 157,785 bp circular dsDNA genome revealed a number of interesting features. ΦSH19 constitutes another member of the recently-proposed Myoviridae Vi01-like family of phages, containing S. Typhi-specific Vi01 and Shigella-specific SboM-AG3. At the nucleotide level ΦSH19 is highly similar to phage Vi01 (80-98% pairwise identity over the length of the genome, with the major differences lying in the region associated with host-range determination. Analyses of the proteins encoded within this region by ΦSH19 revealed a cluster of three putative tail spikes. Of the three tail spikes, two have protein domains associated with the pectate lyase family of proteins (Tsp2 and P22 tail spike family (Tsp3 with the prospect that these enable Salmonella O antigen degradation. Tail spike proteins of Vi01 and SboM-AG3 are predicted to contain conserved right-handed parallel β-helical structures but the internal protein domains are varied allowing different host specificities. Conclusions The addition or exchange of tail spike protein modules is a major contributor to host range determination in the Vi01-like phage family.
Sílvia Dias de Oliveira
Full Text Available The presence of three virulence genes, invA, spvR, and spvC, was determined in Salmonella Enteritidis isolated from poultry, pigs, humans and food. All isolates were positive for the invA gene, with 91.2% being positive for spvR and 90.2% for spvC. There was no significant difference in the prevalence of the virulence genes between isolates from different sources. The results indicate that there is a putative high virulence potential for the S. Enteritidis isolates characterized.A presença de três genes de virulência (invA, spvR e spvC foi determinada em Salmonella Enteritidis isoladas de aves, suínos, humanos e alimentos. Todos os isolados foram positivos para o gene invA, 91,2% também foram positivos para o spvR e 90,2% para o spvC. Não existiu diferença significativa na prevalência dos genes de virulência entre isolados de diferentes origens. Os resultados indicaram que, provavelmente, exista um alto potencial de virulência nos isolados de S. Enteritidis caracterizados.
高瑞娟; 张凯; 吕嘉敏; 黄永兴; 罗开健
This study aimed to establish a rapid detection of Campylobacter jejuni and Salmonella by duplex real-time PCR.The primers and Taq man probes were designed based on the invA gene of Salmonella and hipO gene of Campylobacter jejuni ,using FAM,JOE and TAMRA fluorescently labeled specific probes aimed at the conserved genes of target pathogens,respectively.We compared Tm values to fix these primers and probes,and optimize the reaction conditions to establish the duplex real-time PCR approach for the detection of Campylobacter jejuni and Salmonella.The sensitivity of the duplex real-time PCR was 10 CFU/mL,230 CFU/mL for Campylobacterjejuni and Salmonella.The duplex real-time PCR showed good specificity which was validated by testing various bacteria isolates.In this study,a duplex PCR with high specificity and sensitivity was developed and it would provide useful information for the simultaneous detection of Campylobacter jejuni and Salmonella in food.%为建立一种空肠弯曲菌和沙门菌的双重实时荧光定量 PCR 检测方法，根据空肠弯曲菌的保守基因 hipO 和沙门菌的保守基因 invA 分别设计合成引物和 TaqMan 探针，分别使用 FAM、JOE 作为探针报告基团，TAMRA 作为探针淬灭基团。优化反应体系及条件，建立适用于检测食品中空肠弯曲菌和沙门菌的双重实时荧光定量 PCR 方法。结果显示，该检测方法特异性强，灵敏度高，空肠弯曲菌检测限可达10 CFU/mL，沙门菌检测限达230 CFU/mL。表明建立的双重实时荧光定量 PCR 可为实现食品中空肠弯曲菌和沙门菌的同时检测提供新方法。
Hui Zhang; Liu Liu; Ke Wen; Jinlin Huang; Shizhong Geng; Junsong Shen; Zhiming Pan; Xinan Jiao
The flagellin component FliC of Salmonella typhimurium is capable of activating the innate immune system via specific interactions with TLR5 and can also act as a carrier of foreign antigen to elicit antigen-specific immune responses.Thus,we constructed an attenuated Salmonella strain SL5928(fliC/esat) expressing chimeric flagellin that contained the ESAT-6 antigen coding sequence of Mycobacterium tuberculosis inserted into the highly variable region of the Salmonella flagellin coding gene fliCi.The chimeric flagellin functioned normally,as demonstrated using a flagella swarming assay and electron microscopy.To analyze the effects of chimeric flagellin,the cell-mediated immune response and cytotoxic T lymphocyte (CTL) effects specific for ESAT-6antigen were tested after intranasal immunization of mice with flagellated Salmonella SL5928(fliC/esat).The results showed that SL5928(fliC/esat) intranasal immunization can strongly elicit an ESAT-6-specific T helper (Th) 1-type immune response in mucosal lymphoid tissues,such as nasopharynx-associated lymph nodes,lung and Peyer's patches,and a Th 1/Th2 response was elicited in spleen and mesenteric lymph nodes.Furthermore,intranasal immunization of SL5928(fliC/esat) produced efficient CTL effects,as demonstrated using a 5-and 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) assay.Thus,our study revealed that Salmonella flagellin acts as a carrier for foreign antigen and triggers strong Th 1 and CTL responses during intranasal immunization.Chimeric flagellin is potentially an effective strategy for the development of novel vaccines against tuberculosis in humans and animals.
Pui, Chai Fung; Wong, Woan Chwen; Chai, Lay Ching; Lee, Hai Yen; Noorlis, Ahmad; Zainazor, Tuan Chilek Tuan; Tang, John Yew Huat; Ghazali, Farinazleen Mohamad; Cheah, Yoke Kqueen; Nakaguchi, Yoshitsugu; Nishibuchi, Mitsuaki; Radu, Son
Salmonellosis outbreaks involving typhoid fever and human gastroenteritis are important diseases in tropical countries where hygienic conditions are often not maintained. A rapid and sensitive method to detect Salmonella spp., Salmonella Typhi and Salmonella Typhimurium is needed to improve control and surveillance of typhoid fever and Salmonella gastroenteritis. Our objective was the concurrent detection and differentiation of these food-borne pathogens using a multiplex PCR. We therefore designed and optimized a multiplex PCR using three specific PCR primer pairs for the simultaneous detection of these pathogens. The concentration of each of the primer pairs, magnesium chloride concentration, and primer annealing temperature were optimized before verification of the specificity of the primer pairs. The target genes produced amplicons at 429 bp, 300 bp and 620 bp which were shown to be 100% specific to each target bacterium, Salmonella spp., Salmonella Typhi and Salmonella Typhimurium, respectively. PMID:22028607
Vendrell, Alejandrina; Mongini, Claudia; Gravisaco, María José; Canellada, Andrea; Tesone, Agustina Inés; Goin, Juan Carlos; Waldner, Claudia Inés
Primary tumor excision is one of the most widely used therapies of cancer. However, the risk of metastases development still exists following tumor resection. The liver is a common site of metastatic disease for numerous cancers. Breast cancer is one of the most frequent sources of metastases to the liver. The aim of this work was to evaluate the efficacy of the orally administered Salmonella Typhi vaccine strain CVD 915 on the development of liver metastases in a mouse model of breast cancer. To this end, one group of BALB/c mice was orogastrically immunized with CVD 915, while another received PBS as a control. After 24 h, mice were injected with LM3 mammary adenocarcinoma cells into the spleen and subjected to splenectomy. This oral Salmonella-based vaccine produced an antitumor effect, leading to a decrease in the number and volume of liver metastases. Immunization with Salmonella induced an early cellular immune response in mice. This innate stimulation rendered a large production of IFN-γ by intrahepatic immune cells (IHIC) detected within 24 h. An antitumor adaptive immunity was found in the liver and celiac and portal lymph nodes (LDLN) 21 days after oral bacterial inoculation. The antitumor immune response inside the liver was associated with increased CD4+ and dendritic cell populations as well as with an inflammatory infiltrate located around liver metastatic nodules. Enlarged levels of inflammatory cytokines (IFN-γ and TNF) were also detected in IHIC. Furthermore, a tumor-specific production of IFN-γ and TNF as well as tumor-specific IFN-γ-producing CD8 T cells (CD8+IFN-γ+) were found in the celiac and portal lymph nodes of Salmonella-treated mice. This study provides first evidence for the involvement of LDLN in the development of an efficient cellular immune response against hepatic tumors, which resulted in the elimination of liver metastases after oral Salmonella-based vaccination. PMID:26973649
Robertsson, J A; Fossum, C; Svenson, S B; Lindberg, A A
Peripheral blood lymphocytes collected from calves infected experimentally with Salmonella typhimurium (O antigens 4,5,12) or Salmonella sp. serotype dublin (O 9,12) were stimulated with various bacterial cell envelope components, and their [3H]thymidine incorporation was measured. It was found that peripheral blood lymphocytes from infected calves incorporated significantly more [3H]thymidine than peripheral blood lymphocytes from uninfected controls (P values ranged from less than 0.05 to l...
Ziethlow, V; Favre, D; Viret, J-F; Frey, J; Stoffel, M H
Diarrhoea caused by enterotoxigenic Escherichia coli (ETEC) requires adhesion of microorganisms to enterocytes. Hence, a promising approach to immunoprophylaxis is to elicit antibodies against colonisation factor antigens (CFAs). Genes encoding the most prevalent ETEC-specific surface antigens were cloned into Vibrio cholerae and Salmonella vaccine strains. Expression of surface antigens was assessed by electron-microscopy. Whereas negative staining was effective in revealing CFA/I and CS3, but not CS6, immunolabelling allowed identification of all surface antigens examined. The V. cholerae vaccine strain CVD103 did not express ETEC-specific colonisation factors, whereas CVD103-HgR expressed CS3 only. However, expression of both CFA/I and CS3 was demonstrated in Salmonella Ty21a. PMID:18093230
Cerisuelo, A; Marín, C; Sánchez-Vizcaíno, F; Gómez, E A; de la Fuente, J M; Durán, R; Fernández, C
Essential oils (EO) and short-chain fatty acids have potential antimicrobial activity in broilers. This study aimed to investigate the effect of a specific blend of EO and a combination of this blend of EO with sodium-butyrate on growth performance and Salmonella colonization in broilers. A total of 480 one-day-old male broilers were distributed into 5 treatments (8 pens per treatment and 12 birds per pen) and reared during 42 d in experimental conditions. Dietary treatments consisted of the addition of different doses of EO (0 mg/kg, control; 50 mg/kg, EO50 and 100 mg/kg, EO100) or a combination of EO with 1 g/kg of sodium-butyrate (B; EO50 + B, EOB50 and EO100 + B, EOB100) to a basal diet. All birds were orally infected with 10(8) cfu of Salmonella Enteritidis on d 7 of study. Individual BW and feed intake per pen were measured at arrival and on a weekly basis. The prevalence and enumeration of Salmonella in feces was determined per treatment at 72 h postinfection and on d 23 and 37 of study. At slaughter, cecal content and liver samples from 16 birds per treatment were cultured for Salmonella and cecal pH was measured. No differences were observed on growth performance among treatments. All fecal samples analyzed were positive for Salmonella from d 10 to the end of the rearing period. At slaughter, Salmonella contamination (positive samples) in cecum was lower in birds fed EOB50 compared with the other treatments (P < 0.05), whereas birds fed the control diet showed the highest colonization rates. The pH of the cecal content was not different among treatments. Thus, EO or its combination with sodium-butyrate did not affect growth performance. However, a clear effectiveness of these products was observed in Salmonella control, especially when low doses of EO were combined with sodium-butyrate (EOB50). PMID:24604853
Kogut, Michael H.; Swaggerty, Christina L.; Byrd, James Allen; Selvaraj, Ramesh; Arsenault, Ryan J.
Non-typhoidal Salmonella enterica induces an early, short-lived pro-inflammatory response in chickens that is asymptomatic of clinical disease and results in a persistent colonization of the gastrointestinal (GI) tract that transmits infections to naïve hosts via fecal shedding of bacteria. The underlying mechanisms that control this persistent colonization of the ceca of chickens by Salmonella are only beginning to be elucidated. We hypothesize that alteration of host signaling pathways mediate the induction of a tolerance response. Using chicken-specific kinomic immune peptide arrays and quantitative RT-PCR of infected cecal tissue, we have previously evaluated the development of disease tolerance in chickens infected with Salmonella enterica serovar Enteritidis (S. Enteritidis) in a persistent infection model (4–14 days post infection). Here, we have further outlined the induction of an tolerance defense strategy in the cecum of chickens infected with S. Enteritidis beginning around four days post-primary infection. The response is characterized by alterations in the activation of T cell signaling mediated by the dephosphorylation of phospholipase c-γ1 (PLCG1) that inhibits NF-κB signaling and activates nuclear factor of activated T-cells (NFAT) signaling and blockage of interferon-γ (IFN-γ) production through the disruption of the JAK-STAT signaling pathway (dephosphorylation of JAK2, JAK3, and STAT4). Further, we measured a significant down-regulation reduction in IFN-γ mRNA expression. These studies, combined with our previous findings, describe global phenotypic changes in the avian cecum of Salmonella Enteritidis-infected chickens that decreases the host responsiveness resulting in the establishment of persistent colonization. The identified tissue protein kinases also represent potential targets for future antimicrobial compounds for decreasing Salmonella loads in the intestines of food animals before going to market. PMID:27472318
Rabsch, Wolfgang; Voigt, Wolfgang; Reissbrodt, Rolf; Tsolis, Renée M.; Bäumler, Andreas J.
Salmonella typhimurium possesses two outer membrane receptor proteins, IroN and FepA, which have been implicated in the uptake of enterobactin. To determine whether both receptors have identical substrate specificities, fepA and iroN mutants and a double mutant were characterized. While both receptors transported enterobactin, the uptake of corynebactin and myxochelin C was selectively mediated by IroN and FepA, respectively.
Clark, M. A.; Reed, K A; Lodge, J; Stephen, J.; Hirst, B H; Jepson, M A
We have examined the role of the Salmonella typhimurium inv locus in invasion of the murine intestine. Previous studies have demonstrated that M cells within the lymphoid-follicle-associated epithelia are the primary site of intestinal invasion by S. typhimurium. In this study, we show that mutants possessing defects in one of two inv genes, invA or invG, which render them severely deficient for invasion of polarized epithelial MDCK cells, retain their ability to actively invade mouse Peyer's...
Jason T Noel
Full Text Available BACKGROUND: Recent outbreaks of vegetable-associated gastroenteritis suggest that enteric pathogens colonize, multiply and persist in plants for extended periods of time, eventually infecting people. Genetic and physiological pathways, by which enterics colonize plants, are still poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: To better understand interactions between Salmonella enterica sv. Typhimurium and tomatoes, a gfp-tagged Salmonella promoter library was screened inside red ripe fruits. Fifty-one unique constructs that were potentially differentially regulated in tomato relative to in vitro growth were identified. The expression of a subset of these promoters was tested in planta using recombinase-based in vivo expression technology (RIVET and fitness of the corresponding mutants was tested. Gene expression in Salmonella was affected by fruit maturity and tomato cultivar. A putative fadH promoter was upregulated most strongly in immature tomatoes. Expression of the fadH construct depended on the presence of linoleic acid, which is consistent with the reduced accumulation of this compound in mature tomato fruits. The cysB construct was activated in the fruit of cv. Hawaii 7997 (resistant to a race of Ralstonia solanacearum more strongly than in the universally susceptible tomato cv. Bonny Best. Known Salmonella motility and animal virulence genes (hilA, flhDC, fliF and those encoded on the pSLT virulence plasmid did not contribute significantly to fitness of the bacteria inside tomatoes, even though deletions of sirA and motA modestly increased fitness of Salmonella inside tomatoes. CONCLUSIONS/SIGNIFICANCE: This study reveals the genetic basis of the interactions of Salmonella with plant hosts. Salmonella relies on a distinct set of metabolic and regulatory genes, which are differentially regulated in planta in response to host genotype and fruit maturity. This enteric pathogen colonizes tissues of tomatoes differently than plant
Löfström, Charlotta; Hansen, Trine; Maurischat, Sven; Malorny, Burkhard
Salmonella remains one of the most important zoonotic pathogenic bacteria and is the causative agents of salmonellosis. The aim of this article is to give an overview of Salmonella and salmonellosis, starting by describing the characteristics of the microorganism Salmonella, including biochemical...
Elemfareji, Omar Ismail; Thong, Kwai Lin
Members of Salmonella enterica are important foodborne pathogens of significant public health concern worldwide. This study aimed to determine a range of virulence genes among typhoidal (S. typhi) and non-typhoidal (S. enteritidis) strains isolated from different geographical regions and different years. A total of 87 S. typhi and 94 S. enteritidis strains were tested for presence of 22 virulence genes by employing multiplex PCR and the genetic relatedness of these strains was further characterized by REP-PCR. In S. typhi, invA, prgH, sifA, spiC, sopB, iroN, sitC, misL, pipD, cdtB, and orfL were present in all the strains, while sopE, agfC, agfA, sefC, mgtC, and sefD were present in 98.8, 97.7, 90.8, 87.4, 87.4 and 17.2 %, of the strains, respectively. No lpfA, lpfC, pefA, spvB, or spvC was detected. Meanwhile, in S. enteritidis, 15 genes, agfA, agfC, invA, lpfA, lpfC, sefD, prgH, spiC, sopB, sopE, iroN, sitC, misL, pipD, and orfL were found in all S. enteritidis strains 100 %, followed by sifA and spvC 98.9 %, pefA, spvB and mgtC 97.8 %, and sefC 90.4 %. cdtB was absent from all S. enteritidis strains tested. REP-PCR subtyped S. typhi strains into 18 REP-types and concurred with the virulotyping results in grouping the strains, while in S. enteritidis, REP-PCR subtyped the strains into eight profiles and they were poorly distinguishable between human and animal origins. The study showed that S. typhi and S. enteritidis contain a range of virulence factors associated with pathogenesis. Virulotyping is a rapid screening method to identify and profile virulence genes in Salmonella strains, and improve an understanding of potential risk for human and animal infections. PMID:24426144
A. El Allaoui
Full Text Available The present study was conducted to determine the prevalence and the serotypes involved the virulence gene (InvA and SpvC of Salmonella isolates recovered from the raw meat and giblets (liver and gizzard of the turkey in various outlets in the Moroccan market. From November 2011 to November 2012 a total of 192 samples of turkey meat (included 48 breasts, 48 legs, 48 gizzards and 48 livers were collected every ten days from retail outlets in Meknès. Of these, 48 were from popular market, 48 from artisanal slaughterhouses, 48 from poulterers’shops and 48 from a supermarket at Meknes, Morocco. Of the total of 192 samples examined, 24.5% (47/192 were contaminated with Salmonella. Out of the total 48 samples analysed from popular market, 19 (40.42% proved to be Salmonella positive whereas from 48 samples obtained from traditional slaughterhouses and 48 from poulterers’shops 14 (29.87% and 8 (17% contained Salmonella, respectively. Compared to other outlets, a low level of Salmonella contamination was found in samples obtained from Supermarket 6 (12.7%. Among the 47 Salmonella isolates, 6 different serotypes were identified of which S. Saintpaul (46.8% was the most frequent, followed by S. Agona (17% and S. Kentucky (17%, S. Typhimurium (8.5%, S. Infantis (6.3% and S. Bredeney (4.2%.The high level of contamination, especially in popular market and artisanal slaughterhouses of turkey meat and giblets with Salmonella observed in this paper indicates the need for an improvement in the microbiological quality of retail turkey. Examination of Salmonella for invA gene was detected in all the strains (n=47, only three isolates were positive for the gene SpvC: S. Agona, S. Kentucky and S. Infantis.
Schindler, Peter R. G.; Teuber, Michael
Amino groups of cell envelope proteins, lipids, and lipopolysaccharides cannot be labeled in intact cells of Salmonella typhimurium G 30 by using 5-dimethylaminonaphthalene-1-sulfonylchloride incorporated in lecithin-cholesterol vesicles. However, application of membrane-interacting agents like tris(hydroxymethyl)aminomethane (Tris)-hydrochloride, ethylenediaminetetraacetate (Na salt) (EDTA), divalent cations, and sublethal doses of the cationic antibacterial agents polymyxin B and chlorhexidine induced specific fluorescent labeling of envelope proteins and lipids but not of cytoplasmic compounds, with the exception of a soluble protein with a molecular weight of 46,000 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Treatment with Tris-hydrochloride buffer produced labeling of the heat-modifiable protein B/B+ and of proteins with molecular weights of 26,000, 22,000, and below 17,000. A combination of Tris-hydrochloride and EDTA induced additional dansylation of the major protein A and of proteins of molecular weights 80,000, 60,000, and 44,000. Polymyxin B and chlorhexidine caused similar labeling patterns. In every case, except with divalent cation treatment, protein B/B+ was the most prominently labeled species. Phosphatidylethanolamine was dansylated up to 30%. Lipopolysaccharide was not reactive under any condition or treatment. In addition, the peptidoglycan-bound lipoprotein did not react with dansylchloride in either intact or Tris-hydrochloride-treated cells. The results are discussed with regard to a possible localization of labeled and unlabeled compounds of the cell envelope on the basis of a model placing cell envelope amino groups into ion-ion interactions with anionic components of other envelope compounds like phosphate and carboxyl groups. Images PMID:97268
Prentice, Michael B.; James, Keith D.; Parkhill, Julian; Baker, Stephen G.; Stevens, Kim; Simmonds, Mark N.; Mungall, Karen L.; Churcher, Carol; Oyston, Petra C. F.; Titball, Richard W.; Wren, Brendan W.; Wain, John; Pickard, Derek; Hien, Tran Tinh; Farrar, Jeremy J.; Dougan, Gordon
Population genetic studies suggest that Yersinia pestis, the cause of plague, is a clonal pathogen that has recently emerged from Yersinia pseudotuberculosis. Plasmid acquisition is likely to have been a key element in this evolutionary leap from an enteric to a flea-transmitted systemic pathogen. However, the origin of Y. pestis-specific plasmids remains obscure. We demonstrate specific plasmid rearrangements in different Y. pestis strains which distinguish Y. pestis bv. Orientalis strains from other biovars. We also present evidence for plasmid-associated DNA exchange between Y. pestis and the exclusively human pathogen Salmonella enterica serovar Typhi. PMID:11274119
Of more than 2500 identified Salmonella serotypes, only a small proportion are common in poultry flocks. However, there is an epidemiologically important connection between poultry products and human infections, as many of the serotypes that are most prevalent in humans (such as S. Typhimurium and S...
V. K. Naik
Full Text Available Aim: The aim was to assess the prevalence of Salmonella in raw chevon and chicken meat sold in the retail meat shops situated in and around Durg, Rajnandgaon, Dhamtari, Raipur, and Bilaspur districts of Chhattisgarh. Studies were also conducted to find out the antibiotic resistance in Salmonella isolates. Materials and Methods: A total of 400 samples comprising of 200 chevon meat and 200 chicken meat samples were processed for isolation of Salmonella and all isolates were further confirmed on the basis of cultural and biochemical characters and by targeting invA gene of Salmonella. All Salmonella isolates were also examined for their antimicrobial drug susceptibility/resistance pattern against commonly used antibiotics. Results: Out of 400 samples, the prevalence of Salmonella in chevon and chicken meat was found 9% and 7% respectively, with an overall prevalence of 8%. Polymerase chain reaction targeting invA gene of Salmonella showed positive result with 31 isolates. All 32 Salmonella isolates were found to be highly sensitive to ciprofloxacin while 96.87%, 96.87% and 93.75% were sensitive to gentamicin, imipenem, and ceftazidime, respectively. 93.75% and 59.37% isolates were resistant to erythromycin and oxytetracycline, respectively. Out of 32, 14 isolates had multiple antibiotic resistance index equal to or more than 0.2. Conclusion: Salmonella in chevon and chicken meat samples is prevailing in the areas of sampling due to poor hygienic conditions and also demonstrated the varied spectrum of antimicrobial resistance, including several multiple drug resistance phenotypes. Therefore, the present study emphasizes the need for continued surveillance of zoonotic foodborne pathogens including antimicrobial-resistant variants throughout the food production chain.
Zhai, Ligong; Kong, Xiaohan; Lu, Zhaoxin; Lv, Fengxia; Zhang, Chong; Bie, Xiaomei
S. Dublin has caused widespread concerns in cattle produce. Using a comparative genomic method, two specific targets like SeD_A1118 and SeD_A2283 for S. Dublin identification were firstly obtained. An efficient multiplex PCR for S. Dublin detection based on the two novel specific genes and invA was therefore developed. PMID:24607499
. However, one new clone of multi-resistant Salmonella Kentucky was found. This study provided us with new insights into the genetic diversity on NTS in Senegal. Molecular tools remain essential to study the epidemiology of NTS by tracking the sources of infection and/or contamination. These same techniques were used to study the animal to human transmission in The Gambia in the next chapter. In chapter 4, eight diarrheic children with confirmed salmonellosis and 6 healthy carriers were traced back to their compounds and Salmonella identified from the domestic animals (poultry, sheep and goat living in close contact in the same compound. The most common serotypes identified were Salmonella Colindale in humans (21.42% and Salmonella Poona in animals (14.28%. Among the animals, poultry carried the highest proportion of Salmonella (66.7%. In fact, poultry are considered as the most common asymptomatic carriers of Salmonella. However, serotypes in humans were different from those in animals except in one case where Salmonella Moualine was simultaneously found in chicken and a diarrheic child but in different compounds. After proceeding MLST on all isolates, we found that those two Salmonella Moualine were distinct but genetically very close because they differed at only one locus sucA. The similarity matrix of the strains revealed close genetic relatedness among Salmonella serotypes. There was at least 80% similarity and the majority varied between 98% and 100%. This showed the stability of Salmonella clones which are not subject to high genetic variability. There was therefore no indication of clonal groups which are adapted to a specific host because the genetic tree did reveal that all lineages contained isolates of mixed origin (human and animal. The association between salmonellosis and other diseases, most often malaria, in our study shows the role of opportunistic infections and malaria in NTS infections. Almost all serotypes were susceptible to all antibiotics tested
Song, Saemee; Hwang, Soonhye; Lee, SeungHwa; Ha, Nam-Chul; Lee, Kangseok
To survive in the presence of a wide range of toxic compounds, gram-negative bacteria expel such compounds via tripartite efflux pumps that span both the inner and outer membranes. The Salmonella-specific MdsAB pump consists of MdsB, a resistance-nodulation-division (RND)-type inner membrane transporter (IMT) that requires the membrane fusion protein (MFP) MdsA, and an outer membrane protein (OMP; MdsC or TolC) to form a tripartite efflux complex. In this study, we investigated the role of th...
Three out of five rabbits subjected to successive irradiations and immunized against Salmonella abortus-equi produced IgG which, transiently, were not precipitated by anti-a3 anti-allotypic sera, although they carried all the determinants of the a3 allotypic pattern. As a3 IgG from normal rabbits are precipitated by anti-a3 sera, it appeared that the molecular distribution of the a3 allotypic determinants was different on the IgG produced by these irradiated rabbits compared to IgG produced by normal rabbits. After a second irradiation, one of these rabbits produced a high level (50 mg per ml) of anti-Salmonella antibodies of restricted heterogeneity. An anti-idiotypic serum prepared against anti-Salmonella antibodies produced by this rabbit after the first irradiation precipitated the idiotypes it recognised in the serum collected after the first irradiation, while it did not precipitate the idiotypes it recognised in the serum collected after the second irradiation, although they carried all the determinants of the idiotypes of the serum collected after the first irradiation. That probably means that there is a different molecular distribution of the idiotypic determinants between antibodies produced after the first irradiation and antibodies produced after the second irradiation. An antiidiotypic serum prepared against anti-Salmonella antibodies produced after the second irradiation did not distinguish by precipitation in gel medium or by radioimmunoassay between the idiotypes it recognised in antibodies produced after the first irradiation and those in antibodies produced after the second irradiation. The idiotypic similarity thus detected, and the fact that unexpected allotypes were not detected, is in better agreement with the expression of the potentiality of radiation-resistant cells than with the expression of new antibody producing cell clones arising from virgin stem cells. (author)
Bej, A K; Mahbubani, M H; Boyce, M. J.; Atlas, R M
PCR DNA amplification of a region of the himA gene of Salmonella typhimurium specifically detected Salmonella spp. In oysters, 1 to 10 cells of Salmonella spp. were rapidly detected by the PCR following a pre-enrichment step to increase sensitivity and to ensure that detection was based on the presence of viable Salmonella spp.
Kim, Hyun-Joong; Park, Si-Hong; Kim, Hae-Yeong
Genes of Salmonella enterica serovar Typhimurium LT2 expected to be specifically present in Salmonella were selected using the Basic Local Alignment Search Tool (BLAST) program. The 152 selected genes were compared with 11 genomic sequences of Salmonella serovars, including Salmonella enterica subsp. I and IIIb and Salmonella bongori (V), and were clustered into 17 groups by their comparison patterns. A total of 38 primer pairs were constructed to represent each of the 17 groups, and PCR was ...
Nyman, Ann-Kristin J; Ågren, Estelle CC; Bergström, Karin; Wahlström, Helene
Background The Swedish Salmonella control program has been running for decades and has resulted in a low prevalence of Salmonella in Swedish food producing animals. Routine bacteriology is used to detect Salmonella, however, bacteriology is time consuming, costly and has a low sensitivity. Different enzyme-linked immunosorbent assays (ELISAs) have been developed for detection of antibodies against Salmonella Dublin and S. Typhimurium in bovine bulk milk, individual milk samples as well as in ...
Kim, Ji-Hyun; Rhim, Seong-Ryul; Kim, Kee-Tae; Paik, Hyun-Dong
A rapid and specific PCR assay for the simultaneous detection of Listeria monocytogenes, Escherichia coli O157:H7, Bacillus cereus, Salmonella spp., and Staphylococcus aureus in foods was developed to reduce the detection time and to increase sensitivity. Multiplex PCR developed in this study produced only actA, fliC, hbl, invA, ileS amplicons, but did not produce any non-specific amplicon. The primer sets successfully amplified the target genes in the multiplex PCR without any non-specific or additional bands on the other strains. The multiplex PCR assays also amplified some target genes from five pathogens, and multiplex amplification was obtained from as little as 1 pg of DNA. According to the results from the sensitivity evaluation, the multiplex PCR developed in this study detected 10 cells/mL of the pathogens inoculated in milk samples, respectively. The results suggested that multiplex PCR was an effective assay demonstrating high specificity for the simultaneous detection of five target pathogens in food system. PMID:26761507
Maria Braukmann; Ulrich Methner; Angela Berndt
Salmonella serovars are differentially able to infect chickens. The underlying causes are not yet fully understood. Aim of the present study was to elucidate the importance of Salmonella Pathogenicity Island 1 and 2 (SPI-1 and -2) for the virulence of two non-host-specific, but in-vivo differently invasive, Salmonella serovars in conjunction with the immune reaction of the host. Primary avian splenic macrophages were inoculated with Salmonella enterica sub-species enterica serovar (S.) Typhim...
Poharkar, Krupali V; Kerkar, Savita; D'Costa, Dilecta; Doijad, Swapnil; Barbuddhe, S B
Mangroves are affected by industrial and anthropogenic factors. Although mangroves have been widely studied, investigations of pathogens that may affect public health significance are largely lacking even while incidences of diseases linked with the consumption of mangrove-associated food have increased. A total of 150 samples of water, sediment, and biota were collected from ten mangrove ecosystems in Goa, India. Total viable counts of pathogens such as E. coli, Listeria, Salmonella, and Vibrio spp. ranged from 1.25 to 3.9 × 10(3) cfu/ mL, which were above the relevant standards. Salmonella counts were the highest at 3.1 to 3.9 × 10(3)cfu/mL, with a prevalence of 40%. Considering its high prevalence, the virulence of Salmonella spp. was studied. The invA gene was detected in 35% of the Salmonella isolates by polymerase chain reaction (PCR). The findings suggested that pathogens adapt to this habitat, resulting in contamination of the indigenous fauna. PMID:26931537
Full Text Available Neonatal calf diarrhea remains one of the most important problems faced by livestock, causing great economic losses. This study investigated the prevalence of Salmonella and Escherichia coli, especially enterotoxigenic E. coli (ETEC, in diarrheic calves. Fecal samples were collected from 127 diarrheic calves up to 3 months of age at 12 farms from different governorates in Egypt. 119 bacterial isolates (93.7% were recovered and the prevalences of Salmonella and E. coli in diarrheic calves were 18.1% and 75.6%, respectively. Serotyping of Salmonella isolates revealed that S. Enteritidis and S. Typhimurium were the most prevalent serotypes, representing 60.9% and 30.4%, respectively, while S. Dublin was 8.7%. Serogrouping of E. coli isolates showed that 10 O-serogroups were obtained where O26 and O103 were the most prevalent (17.7% of each. Salmonella serotypes showed positive results with PCR test using oligonucleotide primer amplifying 521 bp fragment of invA gene of Salmonella while 70% of E. coli serogroups possessed ETEC virulent gene (K99. The in-vitro antibiotic sensitivity test indicated that Salmonella serotypes showed high sensitivity against enrofloxacin, spectinomycin and neomycin while E. coli isolates showed high sensitivities against marbofloxacin, spectinomycin and neomycin only.
... Salmonella Reading and Salmonella Abony Infections linked to Alfalfa Sprouts Advice to Consumers & Retailers Case Count Maps ... Signs & Symptoms Key Resources Salmonella Infections Linked to Alfalfa Sprouts from One Contaminated Seed Lot Advice to ...
... What's this? Submit Button Past Emails CDC Features Reptiles, Amphibians, and Salmonella Language: English Español (Spanish) Recommend ... live. How do people get Salmonella infections from reptiles and amphibians? Reptiles and amphibians might have Salmonella ...
Bogomolnaya Lydia M
Full Text Available Abstract Background Salmonellosis is one of the most important bacterial food borne illnesses worldwide. A major source of infection for humans is consumption of chicken or egg products that have been contaminated with Salmonella enterica serotype Typhimurium, however our knowledge regarding colonization and persistence factors in the chicken is small. Results We compared intestinal and systemic colonization of 1-week-old White Leghorn chicks and Salmonella-resistant CBA/J mice during infection with Salmonella enterica serotype Typhimurium ATCC14028, one of the most commonly studied isolates. We also studied the distribution of wild type serotype Typhimurium ATCC14028 and an isogenic invA mutant during competitive infection in the cecum of 1-week-old White Leghorn chicks and 8-week-old CBA/J mice. We found that although the systemic levels of serotype Typhimurium in both infected animal models are low, infected mice have significant splenomegaly beginning at 15 days post infection. In the intestinal tract itself, the cecal contents are the major site for recovery of serotype Typhimurium in the cecum of 1-week-old chicks and Salmonella-resistant mice. Additionally we show that only a small minority of Salmonellae are intracellular in the cecal epithelium of both infected animal models, and while SPI-1 is important for successful infection in the murine model, it is important for association with the cecal epithelium of 1-week-old chicks. Finally, we show that in chicks infected with serotype Typhimurium at 1 week of age, the level of fecal shedding of this organism does not reflect the level of cecal colonization as it does in murine models. Conclusion In our study, we highlight important differences in systemic and intestinal colonization levels between chick and murine serotype Typhimurium infections, and provide evidence that suggests that the role of SPI-1 may not be the same during colonization of both animal models.
Campos-Galvão, Maria Emilene Martino; Ribon, Andrea Oliveira Barros; Araújo, Elza Fernandes; Vanetti, Maria Cristina Dantas
Quorum sensing is used by bacteria to coordinate gene expression in response to population density and involves the production, detection and response to extracellular signaling molecules known as autoinducers (AIs). Salmonella does not synthesize the AI-1, acyl homoserine lactone (AHL) common to gram-negative bacteria; however, it has a receptor for AI-1, the SdiA protein. The effect of SdiA in modulating phenotypes of Salmonella has not been elucidated. In this report, we provide evidence that the AIs-1 affect Salmonella enterica serovar Enteritidis behavior by enhancing the biofilm formation and expression of virulence genes under anaerobic conditions. Biofilm formation by Salmonella was detected by the crystal violet method and by scanning electron microscopy. The presence of AHLs, particularly C12-HSL, increased biofilm formation and promoted expression of biofilm formation genes (lpfA, fimF, fliF, glgC) and virulence genes (hilA, invA, invF). Our results demonstrated that AHLs produced by other organisms played an important role in virulence phenotypes of Salmonella Enteritidis. PMID:26662614
Lundin, B Samuel; Johansson, Camilla; Svennerholm, Ann-Mari
The kinetics and homing characteristics of T-cell responses in humans after mucosal immunizations have not been well characterized. Therefore, we have investigated the magnitude and duration of such responses as well as the homing receptor expression of antigen-specific peripheral blood T cells by using an oral model vaccine, i.e., the live, attenuated Salmonella enterica serovar Typhi vaccine (Ty21a). Eight volunteers were each given three doses of the vaccine 2 days apart, and blood samples, from which CD4(+) and CD8(+) T cells were selected by the use of magnetic beads, were collected before vaccination and at regular intervals thereafter. To purify the potentially antigen-specific gut-homing T cells, CD45RA(-) integrin beta(7)(+) cells were further sorted by flow cytometry. The sorted cells were then stimulated in vitro with the serovar Typhi vaccine strain, and the proliferation of cells and the cytokine production were measured. Following vaccination, there was a large increase in both the proliferation of and the gamma interferon (IFN-gamma) production by blood T cells stimulated with the vaccine strain. The responses were seen among both CD4(+) and CD8(+) T cells, although the CD8(+) cells produced the largest amounts of IFN-gamma. Peak responses were seen 7 to 14 days after the onset of vaccination. Furthermore, most of the IFN-gamma produced by both CD4(+) and CD8(+) cells emanated from cells with the potential to home to mucosal tissues, as the integrin beta(7)-expressing memory T cells produced around 10-fold more IFN-gamma than the remaining populations. In conclusion, we demonstrate that oral vaccination with a live oral bacterial vaccine induces antigen-specific CD4(+) and CD8(+) memory T cells, almost all of which express the gut-homing integrin beta(7). PMID:12228290
Full Text Available Abstract Background Recombinant Salmonella vaccine vectors may potentially be used to induce specific CD4+ T cell responses against foreign viral antigens. Such immune responses are required features of vaccines against pathogens such as human immunodeficiency virus type 1 (HIV-1. The aim of this study was to investigate the induction of systemic HIV-1-specific CD4+ T helper (Th responses in mice after oral immunization with a live attenuated Salmonella vaccine vector that expressed HIV-1 subtype C Gag. Groups of BALB/c mice were vaccinated orally three times (4 weeks apart with this recombinant Salmonella. At sacrifice, 28 days after the last immunization, systemic CD4+ Th1 and Th2 cytokine responses were evaluated by enzyme-linked immunospot assay and cytometric bead array. HIV-1 Gag-specific IgG1 and IgG2a humoral responses in the serum were determined by enzyme-linked immunosorbent assay. Results Mice vaccinated with the recombinant Salmonella elicited both HIV-1-specific Th1 (interferon-gamma (IFN-γ and tumour necrosis factor-alpha (TNF-α and Th2 (interleukin-4 (IL-4 and interleukin-5 (IL-5 cytokine responses. The vaccine induced 70 (IFN-γ spot-forming units (SFUs/10e6 splenocytes and 238 IL-4 SFUs/10e6 splenocytes. Splenocytes from vaccinated mice also produced high levels of Th1 and Th2 cytokines upon stimulation with a Gag CD4 peptide. The levels of IFN-γ, TNF-α, IL-4 and IL-5 were 7.5-, 29.1-, 26.2- and 89.3-fold above the background, respectively. Both HIV-1 Gag-specific IgG1 and IgG2a antibodies were detected in the sera of vaccinated mice. Conclusion The study highlights the potential of orally-delivered attenuated Salmonella as mucosal vaccine vectors for HIV-1 Subtype C Gag to induce Gag-specific CD4+ Th1 and Th2 cellular immune responses and antibodies which may be important characteristics required for protection against HIV-1 infection.
Mercanoğlu, Birce; Aytaç, S. Aykut
In this study, firstly the specificity of Dynabeads® antiâ'Salmonella has been examined and then (immunomagnetic separation) IMS method has been investigated in order to isolate Salmonella spp. from 50 food samples assumed that having the contamination of Salmonella risk in parallel with the traditional cultural method that is ISO 6579 reference method. Also Salmonella enteritidis, antiâ'Salmonella beads and the S. enteritidis +Dynabeads® antiâ'Salmonella complex...
Osman, K M; Marouf, S H; Erfan, A M; AlAtfeehy, N
Globalisation and international trade facilitate the rapid spread and transmission of foodborne pathogens. This study was designed to determine the serovars, distribution of virulence genes (invA, avrA, ssaQ, mgtC, siiD, sopB, gipA, sodC1, sopE1, spvC, bcfC) and antibiotic resistance profiles in salmonellae recovered from imported and domestic day-old turkey poults in Egypt. The prevalence of salmonellae in the imported poults was 4% (6/150): S. Enteritidis was the most frequent isolate (1.3%; 2/150), followed by Typhimurium, Virchow, Larochelle and a non-typeable strain, each with 0.7% (1/150) prevalence. The prevalence of salmonellae in the domestic poults was < 2% (2/150) and serotyping indicated a prevalence of 1.3% (1/150) for both Typhimurium and Altona. In polymerase chain reaction screening, the genes invA, sopB and bcfC were detected in all the Enteritidis, Typhimurium, Virchow, Larochelle, Altona and non-typeable isolates (100%); the gene gipA was absent from all isolates. Carriage of invA, sopB and bcfC among the Enteritidis, Typhimurium, Virchow, Larochelle, Altona and non-typeable isolates was associated with a core pattern of resistance to three antibiotics: streptomycin, nalidixic acid and chloramphenicol. The detection of S. Enteritidis, Typhimurium, Virchow, Larochelle, and Altona in turkey poults has important implications because these serovars are a significant cause of foodborne illness and enteric fever in humans. PMID:25812224
Jensen, Annette Nygaard; Hoorfar, Jeffrey
A rapid biochemical system (OBIS) based on immediate enzymatic differentiation of Citrobacter, Proteus, Providencia, Hafnia and Morganella spp. from Salmonella on brilliant green agar was evaluated A total of 96 field isolates from various Salmonella serotypes, 18 Citrobacter freundii and 25...... isolates of other Enterobacteriaceae were tested All Salmonella isolates were identified correctly by the kit, and none of the Enterobacteriaceae isolates were identified as Salmonella. The results indicate complete specificity for Salmonella colonies on brilliant green agar....
Abstract Benzidine, 4-aminobiphenyl, 3,3'-dichlorobenzidine HCl, 3,3'-dimethylbenzidine, 3,3'- dimethoxybenzidine and benzidine congener-based dye trypan blue were mutagenic in Salmonella typhimurium TAl 00 only with metabolic activation. It was found that a hamster liver 89 ...
Salmonella spp., Listeria monocytogenes, and Escherichia coli O157:H7 are food borne-pathogens capable of causing serious gastrointestinal illness. We previously described simultaneous detection of these pathogens by multiplex PCR in 44 types of spiked food samples, including meat, produce, fish, an...
Oliver T. Zishiri
Full Text Available Salmonellosis is a significant public health concern around the world. The injudicious use of antimicrobial agents in poultry production for treatment, growth promotion and prophylaxis has resulted in the emergence of drug resistant strains of Salmonella. The current study was conducted to investigate the prevalence of virulence and antimicrobial resistance genes from Salmonella isolated from South African and Brazilian broiler chickens as well as human clinical isolates. Out of a total of 200 chicken samples that were collected from South Africa 102 (51% tested positive for Salmonella using the InvA gene. Of the overall 146 Salmonella positive samples that were screened for the iroB gene most of them were confirmed to be Salmonella enterica with the following prevalence rates: 85% of human clinical samples, 68.6% of South African chicken isolates and 70.8% of Brazilian chicken samples. All Salmonella isolates obtained were subjected to antimicrobial susceptibility testing with 10 antibiotics. Salmonella isolates from South African chickens exhibited resistance to almost all antimicrobial agents used, such as tetracycline (93%, trimethoprim-sulfamthoxazole (84%, trimethoprim (78.4%, kanamycin (74%, gentamicin (48%, ampicillin (47%, amoxicillin (31%, chloramphenicol (31%, erythromycin (18% and streptomycin (12%. All samples were further subjected to PCR in order to screen some common antimicrobial and virulence genes of interest namely spiC, pipD, misL, orfL, pse-1, tet A, tet B, ant (3"-la, sul 1 and sul. All Salmonella positive isolates exhibited resistance to at least one antimicrobial agent; however, antimicrobial resistance patterns demonstrated that multiple drug resistance was prevalent. The findings provide evidence that broiler chickens are colonised by pathogenic Salmonella harbouring antimicrobial resistance genes. Therefore, it is evident that there is a need for prudent use of antimicrobial agents in poultry production systems in
... HUMAN SERVICES Food and Drug Administration Salmonella Contamination of Dry Dog Food; Withdrawal of...) entitled ``Sec. 690.700 Salmonella Contamination of Dry Dog Food.'' This CPG is obsolete. DATES: The.... SUPPLEMENTARY INFORMATION: FDA issued the CGP entitled ``Sec. 690.700 Salmonella Contamination of Dry Dog...
Pullinger, Gillian D.; Dziva, Francis; Charleston, Bryan; Wallis, Timothy S.; Stevens, Mark P.
Salmonella enterica serovar Dublin is a host-restricted serovar associated with typhoidal disease in cattle. In contrast, the fowl-associated serovar S. enterica serovar Gallinarum is avirulent in calves, yet it invades ileal mucosa and induces enteritis at levels comparable to those induced by S. enterica serovar Dublin. Suppression subtractive hybridization was employed to identify S. enterica serovar Dublin strain SD3246 genes absent from S. enterica serovar Gallinarum strain SG9. Forty-on...
Elliott, Geoffrey N.; Nadine Thomas; Marion Macrae; Campbell, Colin D.; Ogden, Iain D; Singh, Brajesh K.
A multiplex T-RFLP test was developed to detect and identify Salmonella enterica and all six species of Listeria inoculated into milk at minimal levels. Extensive in silico analysis was used to design a fifteen-primer, six-amplimer methodology and in vitro application showed target organism DNA, when amplified individually, yielded the predicted terminal restriction fragments (TRFs) following digestion. Non-target organisms were either not-amplified or yielded TRFs which did not interfere wit...
Mitchell B. Lerner
Full Text Available Antibody-functionalized carbon nanotube devices have been suggested for use as bacterial detectors for monitoring of food purity in transit from the farm to the kitchen. Here we report progress towards that goal by demonstrating specific detection of Salmonella in complex nutrient broth solutions using nanotube transistors functionalized with covalently-bound anti-Salmonella antibodies. The small size of the active device region makes them compatible with integration in large-scale arrays. We find that the on-state current of the transistor is sensitive specifically to the Salmonella concentration and saturates at low concentration (<1000 cfu/ml. In contrast, the carrier mobility is affected comparably by Salmonella and other bacteria types, with no sign of saturation even at much larger concentrations (108 cfu/ml.
Papasian, C J; Bartholomew, W R; Neter, E.; Amsterdam, D
A patient with a dual Salmonella infection is described. Salmonella group B was recovered from three blood culture sets but was not detected in seven stool cultures. Salmonella group C2 was isolated from three of seven stool cultures but was not recovered from blood cultures. Specific, non-cross-reactive antibodies to Salmonella groups B and C2 were detected in the sera of the patient by passive hemagglutination assays.
Bouvet, P J; Jeanjean, S.
Two colored latex kits (the Wellcolex Colour Salmonella Test [WCT-Salmonella] and the Wellcolex Colour Shigella Test [WCT-Shigella]; Division Diagnostics, Laboratories Wellcome S.A., Paris, France), which allow identification of the most frequently encountered Salmonella serogroups and Shigella species, respectively, were evaluated. WCT-Salmonella and WCT-Shigella yielded sensitivities of 98.4 and 98%, respectively, and a specificity of 100% when they were tested on pure cultures received at ...
Duarte Martínez, Francisco; Sánchez-Salazar, Luz Marina; Acuña-Calvo, María Teresa; Bolaños-Acuña, Hilda María; Dittel-Dittel, Isis; Campos-Chacón, Elena
Salmonella is a complex bacterial group with more than 2400 serovars widely distributed in nature; they are considered zoonotic because they can infect a variety of animals and be transmitted to humans. Usually, they cause alimentary acquired diseases such as gastroenteritis, typhoid fever, and others that can lead to severe complications and death. Serotyping is useful to differentiate among Salmonella, because it shows an important correlation with their clinical and epidemiological patterns; consequently, it is of high value for public health, animal health, agriculture, and industry. To characterize all known Kauffmann-White Salmonella serovars, over 250 antisera are required. Due to this and to high prices antisera, many laboratories worldwide have limitations in establishing Salmonella surveillance. Therefore, we developed and validated a Salmonella flagella microagglutination test (SALMATcor) that significantly reduces laboratory requirements of antisera. SALMATcor is based on scaling down, by fivefold, the antigen:antiserum volumes actually required for the reference method: flagella standard tube agglutination technique (STAT). Antigen preparation, temperatures, and incubation periods remained as established for STAT. The SALMATcor was validated according to ISO/DIS 16140:1999 protocol, which included 1187 comparisons of flagella determinations conducted by SALMATcor and STAT, on 141 Salmonella isolates of 12 common serotypes and the use of antiserum recommended for STAT. SALMATcor concordance was excellent (Cohen's kappa index 0.9982), obtaining relative accuracy >99.9% and relative specificity >99.9%. Additionally, SALMATcor has been used by CNRB-INCIENSA since 2004 to respond to all 40 Salmonella proficiency testing strains, provided by World Health Organization-Global Salmonella Surveillance Network, obtaining 100% concordance on serovar identification. On the basis of the results achieved with SALMATcor and considering that it also significantly
Zishiri, Oliver T; Mkhize, Nelisiwe; Mukaratirwa, Samson
Salmonellosis is a significant public health concern around the world. The injudicious use of antimicrobial agents in poultry production for treatment, growth promotion and prophylaxis has resulted in the emergence of drug resistant strains of Salmonella. The current study was conducted to investigate the prevalence of virulence and antimicrobial resistance genes from Salmonella isolated from South African and Brazilian broiler chickens as well as human clinical isolates. Out of a total of 200 chicken samples that were collected from South Africa 102 (51%) tested positive for Salmonella using the InvA gene. Of the overall 146 Salmonella positive samples that were screened for the iroB gene most of them were confirmed to be Salmonella enterica with the following prevalence rates: 85% of human clinical samples, 68.6% of South African chicken isolates and 70.8% of Brazilian chicken samples. All Salmonella isolates obtained were subjected to antimicrobial susceptibility testing with 10 antibiotics. Salmonella isolates from South African chickens exhibited resistance to almost all antimicrobial agents used, such as tetracycline (93%), trimethoprim-sulfamthoxazole (84%), trimethoprim (78.4%), kanamycin (74%), gentamicin (48%), ampicillin (47%), amoxicillin (31%), chloramphenicol (31%), erythromycin (18%) and streptomycin (12%). All samples were further subjected to PCR in order to screen some common antimicrobial and virulence genes of interest namely spiC, pipD, misL, orfL, pse-1, tet A, tet B, ant (3")-la, sul 1 and sul. All Salmonella positive isolates exhibited resistance to at least one antimicrobial agent; however, antimicrobial resistance patterns demonstrated that multiple drug resistance was prevalent. The findings provide evidence that broiler chickens are colonised by pathogenic Salmonella harbouring antimicrobial resistance genes. Therefore, it is evident that there is a need for prudent use of antimicrobial agents in poultry production systems in order to
Gurman, P M; Ross, T; Holds, G L; Jarrett, R G; Kiermeier, A
Predictive models, to estimate the reduction in Escherichia coli O157:H7 concentration in beef burgers, have been developed to inform risk management decisions; no analogous model exists for Salmonella spp. in pork burgers. In this study, "Extra Lean" and "Regular" fat pork minces were inoculated with Salmonella spp. (Salmonella 4,,12,i:-, Salmonella Senftenberg and Salmonella Typhimurium) and formed into pork burger patties. Patties were cooked on an electric skillet (to imitate home cooking) to one of seven internal temperatures (46, 49, 52, 55, 58, 61, 64 °C) and Salmonella enumerated. A generalised linear logistic regression model was used to develop a predictive model for the Salmonella concentration based on the internal endpoint temperature. It was estimated that in pork mince with a fat content of 6.1%, Salmonella survival will be decreased by -0.2407log10 CFU/g for a 1 °C increase in internal endpoint temperature, with a 5-log10 reduction in Salmonella concentration estimated to occur when the geometric centre temperature reaches 63 °C. The fat content influenced the rate of Salmonella inactivation (P=0.043), with Salmonella survival increasing as fat content increased, though this effect became negligible as the temperature approached 62 °C. Fat content increased the time required for patties to achieve a specified internal temperature (P=0.0106 and 0.0309 for linear and quadratic terms respectively), indicating that reduced fat pork mince may reduce the risk of salmonellosis from consumption of pork burgers. Salmonella serovar did not significantly affect the model intercepts (P=0.86) or slopes (P=0.10) of the fitted logistic curve. This predictive model can be applied to estimate the reduction in Salmonella in pork burgers after cooking to a specific endpoint temperature and hence to assess food safety risk. PMID:26686598
Salmonellosis in Australia has been linked to eggs and egg products with specific serotypes associated with outbreaks. We compared attachment to and survival on egg shells and growth in eggs of two Salmonella serotypes, an egg outbreak associated Salmonella Typhimurium and a non-egg-associated Salmonella enterica ssp. II 1,4,12,27:b:[e,n,x] (S. Sofia). Experiments were conducted at combinations of 4, 15, 22, 37 and 42°C. No significant differences occurred between the serotypes in maximum gro...
Qadri, A; S. K. Gupta; Talwar, G P
Fifteen monoclonal antibodies (MAbs) directed against Salmonella typhi were produced and characterized. The specificities of the antibodies were determined by their binding patterns in an enzyme immunoassay, with a panel of lipopolysaccharides isolated from different bacteria. Seven MAbs reacted with S. typhi, Salmonella enteritidis, and Salmonella dublin (all belonging to serogroup D). One MAb also reacted with Salmonella paratyphi A and S. paratyphi B. Five MAbs reacted with S. typhi, S. en...
Browne, Sara H; Lesnick, Marc L.; Guiney, Donald G.
Infection of human macrophages with Salmonella enterica serovar Typhimurium or Salmonella enterica serovar Dublin produces delayed cytotoxicity characterized by cell detachment and associated apoptosis. Using a site-specific mutant in the SpvB active site, we verify that the ADP-ribosylation activity of SpvB is required for delayed cytotoxicity in human macrophages infected with Salmonella. SipB and the type III protein secretion system (TTSS) encoded by Salmonella pathogenicity island 1 (SPI...
Bula-Rudas, Fernando J; Rathore, Mobeen H; Maraqa, Nizar F
Salmonella are gram-negative bacilli within the family Enterobacteriaceae. They are the cause of significant morbidity and mortality worldwide. Animals (pets) are an important reservoir for nontyphoidal Salmonella, whereas humans are the only natural host and reservoir for Salmonella Typhi. Salmonella infections are a major cause of gastroenteritis worldwide. They account for an estimated 2.8 billion cases of diarrheal disease each year. The transmission of Salmonella is frequently associated with the consumption of contaminated water and food of animal origin, and it is facilitated by conditions of poor hygiene. Nontyphoidal Salmonella infections have a worldwide distribution, whereas most typhoidal Salmonella infections in the United States are acquired abroad. In the United States, Salmonella is a common agent for food-borne–associated infections. Several outbreaks have been identified and are most commonly associated with agricultural products. Nontyphoidal Salmonella infection is usually characterized by a self-limited gastroenteritis in immunocompetent hosts in industrialized countries, but it may also cause invasive disease in vulnerable individuals (eg, children less than 1 year of age, immunocompromised). Antibiotic treatment is not recommended for treatment of mild to moderate gastroenteritis by nontyphoidal Salmonella in immunocompetent adults or children more than 1 year of age. Antibiotic treatment is recommended for nontyphoidal Salmonella infections in infants less than 3 months of age, because they are at higher risk for bacteremia and extraintestinal complications. Typhoid (enteric) fever and its potential complications have a significant impact on children, especially those who live in developing countries. Antibiotic treatment of typhoid fever has become challenging because of the emergence of Salmonella Typhi strains that are resistant to classically used first-line agents: ampicillin, trimethoprim-sulfamethoxazole, and chloramphenicol. The
Mohamed M Ali
Full Text Available Salmonella enterica serovar Typhimurium (Salmonella is one of the most significant food-borne pathogens affecting both humans and agriculture. We have determined that Salmonella encodes an uptake and utilization pathway specific for a novel nutrient, fructose-asparagine (F-Asn, which is essential for Salmonella fitness in the inflamed intestine (modeled using germ-free, streptomycin-treated, ex-germ-free with human microbiota, and IL10-/- mice. The locus encoding F-Asn utilization, fra, provides an advantage only if Salmonella can initiate inflammation and use tetrathionate as a terminal electron acceptor for anaerobic respiration (the fra phenotype is lost in Salmonella SPI1- SPI2- or ttrA mutants, respectively. The severe fitness defect of a Salmonella fra mutant suggests that F-Asn is the primary nutrient utilized by Salmonella in the inflamed intestine and that this system provides a valuable target for novel therapies.
Hoorfar, Jeffrey; Baggesen, Dorte Lau; Porting, P.H.
The purpose of the present study was to investigate the application of ready-to-go Salmonella PCR tests, based on dry chemistry, for final identification of rough presumptive Salmonella isolates. The results were compared with two different biotyping methods performed at two different laboratories......-Salmonella strains, which did not result in any DNA band. A total of 32 out of the 36 rough presumptive isolates were positive in the PCR. All but one isolate were also identified as Salmonella by the two biochemical methods. All 80 Salmonella strains were also tested in the two multiplex serogroup tests based on....... The sensitivity of the BAX Salmonella PCR test was assessed by testing a total of 80 Salmonella isolates, covering most serogroups, which correctly identified all the Salmonella strains by resulting in one 800-bp band in the sample tubes. The specificity of the PCR was assessed using 20 non...
Sharon M Tennant
Full Text Available BACKGROUND: In sub-Saharan Africa, non-typhoidal Salmonella (NTS are emerging as a prominent cause of invasive disease (bacteremia and focal infections such as meningitis in infants and young children. Importantly, including data from Mali, three serovars, Salmonella enterica serovar Typhimurium, Salmonella Enteritidis and Salmonella Dublin, account for the majority of non-typhoidal Salmonella isolated from these patients. METHODS: We have extended a previously developed series of polymerase chain reactions (PCRs based on O serogrouping and H typing to identify Salmonella Typhimurium and variants (mostly I 4,,12:i:-, Salmonella Enteritidis and Salmonella Dublin. We also designed primers to detect Salmonella Stanleyville, a serovar found in West Africa. Another PCR was used to differentiate diphasic Salmonella Typhimurium and monophasic Salmonella Typhimurium from other O serogroup B, H:i serovars. We used these PCRs to blind-test 327 Salmonella serogroup B and D isolates that were obtained from the blood cultures of febrile patients in Bamako, Mali. PRINCIPAL FINDINGS: We have shown that when used in conjunction with our previously described O-serogrouping PCR, our PCRs are 100% sensitive and specific in identifying Salmonella Typhimurium and variants, Salmonella Enteritidis, Salmonella Dublin and Salmonella Stanleyville. When we attempted to differentiate 171 Salmonella Typhimurium (I 4,[ 5],12:i:1,2 strains from 52 monophasic Salmonella Typhimurium (I 4,,12:i:- strains, we were able to correctly identify 170 of the Salmonella Typhimurium and 51 of the Salmonella I 4,,12:i:- strains. CONCLUSION: We have described a simple yet effective PCR method to support surveillance of the incidence of invasive disease caused by NTS in developing countries.
Sutch, K E; Blackburn, B O
Three new Salmonella serotypes belonging to Kauffmann's subgenus I (F. Kauffmann, The Bacteriology of Enterobacteriaceae, 1966) were identified. These serotypes were Salmonella brazos 6,14,18:a:e,n,z15, Salmonella midway 6,14,24:d:1,7, and Salmonella balboa 48a, 48b:z41:monophasic.
Hsieh, Yi-Cheng; Poole, Toni L; Runyon, Mick; Hume, Michael; Herrman, Timothy J
The objective of this study was to characterize 365 nontyphoidal Salmonella enterica isolates from animal feed. Among the 365 isolates, 78 serovars were identified. Twenty-four isolates (7.0%) were recovered from three of six medicated feed types. Three of these isolates derived from the medicated feed, Salmonella Newport, Salmonella Typhimurium var. O 5- (Copenhagen), and Salmonella Lexington var. 15+ (Manila), displayed antimicrobial resistance. Susceptibility testing revealed that only 3.0% (12) of the 365 isolates displayed resistance to any of the antimicrobial agents. These 12 isolates were recovered from unmedicated dry beef feed (n = 3), medicated dry beef feed (n = 3), cabbage culls (n = 2), animal protein products (n = 2), dry dairy cattle feed (n = 1), and fish meal (n = 1). Only Salmonella Newport and Salmonella Typhimurium var. O 5- (Copenhagen) were multidrug resistant. Both isolates possessed the IncA/C replicon and the blaCMY-2 gene associated with cephalosporin resistance. Plasmid replicons were amplified from 4 of 12 resistant isolates. Plasmids (40 kb) were Salmonella Montevideo and Salmonella Kentucky. Conjugation experiments were done using 7 of the 12 resistant isolates as donors. Only Salmonella Montevideo, possessing a plasmid and amplifying IncN, produced transconjugants. Transconjugants displayed the same antimicrobial resistance profile as did the donor isolate. Three isolates that amplified replicons corresponding to IncA/C or IncHI2 did not produce transconjugants at 30 or 37°C. The results of this study suggest that the prevalence of antimicrobial-resistant Salmonella contaminating animal feed is low in Texas. However, Salmonella was more prevalent in feed by-products; fish meal had the highest prevalence (84%) followed by animal protein products (48%). Ten of the 35 feed types had no Salmonella contamination. Further investigation is needed to understand the possible role of specific feed types in the dissemination of antimicrobial
Paião, F.G.; Arisitides, L.G.A.; Murate, L.S.; Vilas-Bôas, G.T.; Vilas-Boas, L.A.; Shimokomaki, M.
The presence of Salmonella in the intestinal tract, on the chickens skin and among their feathers, may cause carcasses contamination during slaughtering and processing and possibly it is responsible by the introduction of this microorganism in the slaughterhouses. A rapid method to identify and monitor Salmonella and their sorovars in farm is becoming necessary. A pre-enriched multiplex polymerase chain reaction (m-PCR) assay employing specific primers was developed and used to detect Salmonella at the genus level and to identify the Salmonella enterica serovar Enteritidis (S. Enteritidis) and Salmonella enterica serovar Typhimurium (S. Typhimurium) in broiler chicken swab samples. The method was validated by testing DNA extract from 90 fresh culture cloacal swab samples from poultry chicken cultured in phosphate buffer peptone water at 37 °C for 18 h. The final results showed the presence of Salmonella spp. in 25% of samples, S. Enteritidis was present in 12% of the Salmonella-positive samples and S. Typhimurium in 3% of the samples. The m-PCR assay developed in this study is a specific and rapid alternative method for the identification of Salmonella spp. and allowed the observation of specific serovar contamination in the field conditions within the locations where these chickens are typically raised. PMID:24159281
Full Text Available Aim: Procalcitonin (PCT as a diagnostic marker for bacteremia and sepsis has been extensively studied. We aimed to study PCT levels in Salmonella infections whether they would serve as marker for early diagnosis in endemic areas to start empiric treatment while awaiting blood culture report. Materials and Methods: BACTEC blood culture was used to isolate Salmonella in suspected enteric fever patients. Serum PCT levels were estimated before starting treatment. Results: In 60 proven enteric fever patients, median value of serum PCT levels was 0.22 ng/ml, values ranging between 0.05 and 4 ng/ml. 95% of patients had near normal or mild increase (<0.5 ng/ml, only 5% of patients showed elevated levels. Notably, high PCT levels were found only in severe sepsis. Conclusion: PCT levels in Salmonella infections are near normal or minimally increased which differentiates it from other systemic Gram-negative infections. PCT cannot be used as a specific diagnostic marker of typhoid.
Native DadB and Alr alanine racemases (M/sub r/ 39,000) from Salmonella typhimurium are proteolyzed at homologous positions by α-chymotrypsin, trypsin, and subtilisin to generate in all cases two nonoverlapping polypeptides of M/sub r/ 28,000 and 11,000. Under nondenaturing conditions, chymotryptic digest results in an associated form of the two fragments which possesses 3% of the original catalytic activity, incorporates 0.76 equiv of the mechanism-based inactivator β-chloro-[14C]-D-alanine, and exhibits a UV circular dichroism profile identical with that of native enzyme. Protein sequence analysis of the denatured chymotryptic fragments indicates the presence of a tetrapeptide interdomain hinge (DadB, residues 254-257; Alr, residues 256-259) that is attacked at both ends during proteolysis. Under the previously employed digest conditions, NaB3H4-reduced DadB holoenzyme is resistant to α-chymotrypsin and trypsin and is labile only toward subtilisin. These data suggest that the hinge structure is essential for a catalytically efficient enzyme species and is sensitive to active site geometry. The sequence at the hinge region is also conserved in alanine racemases from Gram-positive bacteria
NOGUEIRA ROBERTO GOMES; TOSCANO VANESSA GRANATO; ABDALA NITAMAR; STÁVALE JOÃO NORBERTO; MALHEIROS SUZANA M. FLEURY; GABBAI ALBERTO ALAIN
Descrevemos o caso de um paciente com diagnóstico de germinoma nos núcleos da base e tálamo, com invasão do tronco cerebral, ressaltando as características observadas nos exames de tomografia computadorizada e de ressonância magnética, os possíveis diagnósticos diferenciais, e a necessidade da comprovação anátomo-patológica, por tratar-se de localização pouco frequente de um tumor com possibilidade de evolução favorável após tratamento com quimio e radioterapia.
Gomba, Annancietar; Chidamba, Lizyben; Korsten, Lise
Foodborne disease outbreaks involving fresh produce have increased in recent years. The risk of infection from contaminated food is worsened by the increased prevalence of antibiotic-resistant strains. This study evaluated the prevalence of antibiotic resistance in Salmonella isolates (n = 263) from agricultural production systems through to the final packed product. Salmonella isolates were preliminarily identified by matrix-assisted laser desorption ionization-time-of-flight mass spectroscopy (MALDI-TOF MS) and API 20E and identities confirmed by invA gene polymerase chain reaction. Antimicrobial susceptibility was performed with 15 antimicrobial agents using the Kirby-Bauer disk diffusion test. Of the 263 Salmonella isolates assessed, 59.3% were resistant to one or more antimicrobials. The most frequently detected resistance was against chloramphenicol and kanamycin (46.7%), trimethoprim-sulfamethoxazole (28%), and streptomycin (14%), and the less frequently detected resistance was toward ampicillin (1.14%), amikacin (0.76%), and amoxicillin-clavulanic acid (0.38%). Multiple antimicrobial resistance (MAR) (resistance to ≥3 antibiotics) was found in 48.7% (76/156) isolates. The most common MAR phenotype was to chloramphenicol and trimethoprim/sulfamethoxazole-kanamycin (43.6%). Resistance to chloramphenicol, kanamycin, or trimethoprim/sulfamethoxazole was only observed in MAR phenotypes. All isolates were susceptible to ceftiofur, cefoxitin, ceftriaxone, ciprofloxacin, nalidixic acid, gentamicin, and tetracycline. This study confirms the importance of fresh produce production environments as potential reservoirs and fresh produce as carriers of antibiotic-resistant Salmonella spp. with significant clinical importance. Further studies to evaluate the actual level of health risk from these pathogens should include characterization of the antibiotic resistance determinant genes among the isolates. PMID:27294335
Wang, Yi; Wang, Yan; Luo, Lijuan; Liu, Dongxin; Luo, Xia; Xu, Yanmei; Hu, Shoukui; Niu, Lina; Xu, Jianguo; Ye, Changyun
Shigella and Salmonella are frequently isolated from various food samples and can cause human gastroenteritis. Here, a novel multiple endonuclease restriction real-time loop-mediated isothermal amplification technology (MERT-LAMP) were successfully established and validated for simultaneous detection of Shigella strains and Salmonella strains in only a single reaction. Two sets of MERT-LAMP primers for 2 kinds of pathogens were designed from ipaH gene of Shigella spp. and invA gene of Salmonella spp., respectively. Under the constant condition at 63°C, the positive results were yielded in as short as 12 min with the genomic DNA extracted from the 19 Shigella strains and 14 Salmonella strains, and the target pathogens present in a sample could be simultaneously identified based on distinct fluorescence curves in real-time format. Accordingly, the multiplex detection assay significantly reduced effort, materials and reagents used, and amplification and differentiation were conducted at the same time, obviating the use of postdetection procedures. The analytical sensitivity of MERT-LAMP was found to be 62.5 and 125 fg DNA/reaction with genomic templates of Shigella strains and Salmonella strains, which was consist with normal LAMP assay, and at least 10- and 100-fold more sensitive than that of qPCR and conventional PCR approaches. The limit of detection of MERT-LAMP for Shigella strains and Salmonella strains detection in artificially contaminated milk samples was 5.8 and 6.4 CFU per vessel. In conclusion, the MERT-LAMP methodology described here demonstrated a potential and valuable means for simultaneous screening of Shigella and Salmonella in a wide variety of samples. PMID:26697000
Emborg, H-D; Simonsen, J; Jørgensen, C S;
enabled a back-calculation of the annual seroincidence of Salmonella based on measurements of specific antibodies. The aim of the present study was to determine the seroincidence in two convenience samples from 2012 (Danish blood donors, n = 500, and pregnant women, n = 637) and a community-based sample...... of healthy individuals from 2006 to 2007 (n = 1780). The lowest antibody levels were measured in the samples from the community cohort and the highest in pregnant women. The annual Salmonella seroincidences were 319 infections/1000 pregnant women [90% credibility interval (CrI) 210-441], 182/1000 in...
te Pas Marinus FW
Full Text Available Abstract Background Chicken meat and eggs can be a source of human zoonotic pathogens, especially Salmonella species. These food items contain a potential hazard for humans. Chickens lines differ in susceptibility for Salmonella and can harbor Salmonella pathogens without showing clinical signs of illness. Many investigations including genomic studies have examined the mechanisms how chickens react to infection. Apart from the innate immune response, many physiological mechanisms and pathways are reported to be involved in the chicken host response to Salmonella infection. The objective of this study was to perform a meta-analysis of diverse experiments to identify general and host specific mechanisms to the Salmonella challenge. Results Diverse chicken lines differing in susceptibility to Salmonella infection were challenged with different Salmonella serovars at several time points. Various tissues were sampled at different time points post-infection, and resulting host transcriptional differences investigated using different microarray platforms. The meta-analysis was performed with the R-package metaMA to create lists of differentially regulated genes. These gene lists showed many similarities for different chicken breeds and tissues, and also for different Salmonella serovars measured at different times post infection. Functional biological analysis of these differentially expressed gene lists revealed several common mechanisms for the chicken host response to Salmonella infection. The meta-analysis-specific genes (i.e. genes found differentially expressed only in the meta-analysis confirmed and expanded the biological functional mechanisms. Conclusions The meta-analysis combination of heterogeneous expression profiling data provided useful insights into the common metabolic pathways and functions of different chicken lines infected with different Salmonella serovars.
Nguyen, Scott V.; Bono, James L.; Smith, Timothy P. L.; Fields, Patricia I.; Dinsmore, Blake A.; Santovenia, Monica; Kelley, Christy M.; Wang, Rong; Bosilevac, Joseph M.; Harhay, Gregory P.
Salmonella enterica is an important pathogen transmitted by numerous vectors. Genomic comparisons of Salmonella strains from disparate hosts have the potential to further our understanding of mechanisms underlying host specificities and virulence. Here, we present the closed genome and plasmid sequences of 10 Salmonella enterica subsp. enterica serovar Anatum isolates from bovine and human sources. PMID:27257192
Full Text Available Salmonella Choleraesuis, S. Enteritidis and S. Typhimurium are the main pathogens that contaminate animal products and cause human Salmonella food poisoning. To establish the loop-mediated isothermal amplification (LAMP method for the rapid detection of Salmonella and 3 common Salmonella serotypes, inner and outer primer sets targeting Salmonella invE gene and 3 serotype-specific genes fliC, lygD and STM4495 were designed. The LAMP reaction conditions were optimized. The specificity of LAMP primers was identified by testing 10 different bacterial strains including S. Choleraesuis, S. Enteritidis and S. Typhimurium. Take S. Choleraesuis as example, the detection limit of LAMP assay was 1.33 × 101 CFU/mL for bacteria culture and 2.0 × 101 CFU/mL for simulated pork sample. The results show that LAMP is a rapid, sensitive and specific method for Salmonella detection and can be used for the rapid detection of Salmonella in food.
Hofacre, Charles L.; Peter S. Holt; Lee, Margie D.; Susan Sanchez; Joseph Minicozzi; Maurer, John J.
Accurate and fast detection of harmful Salmonella is a major concern of food safety. Common Salmonella serotypes responsible for human associated foodborne outbreaks are S. Enteritidis, S. Hadar, S. Heidelberg, and S. Typhimurium are also commonly isolated from poultry. Serology is commonly used to monitor disease in poultry, therefore application of Salmonella serotype-specific test will have added value in Salmonella surveillance or monitoring vaccine efficacy. Recombinant flagellins were p...
Fenlon, Luke A; Slauch, James M
Salmonella propagates in macrophages to cause life-threatening infections, but the role of neutrophils in combating Salmonella has been controversial. In this issue, Burton et al. (2014) use single cell analyses and modeling to explain the ability of Salmonella to survive in macrophages while being killed by neutrophils. PMID:24439894
Fenlon, Luke A.; Slauch, James M.
Salmonella propagate in macrophages to cause life-threatening infections, but the role of neutrophils in combating Salmonella has been controversial. In this issue, Burton et al. (2013) use single cell analyses and modeling to explain the ability of Salmonella to survive in macrophages, while being killed by neutrophils.
Eduardo Ferrari Marback
Full Text Available Tumores pigmentados localizados sobre o disco óptico são raros e representam desafio diagnóstico. Paciente masculino, 60 anos, apresenta baixa da acuidade visual no olho esquerdo devido à lesão pigmentada que cobre o disco óptico. Foi indicada a enucleação com recusa pelo paciente. O quadro evoluiu com descolamento de retina. Examinado em outro serviço teve indicação de vitrectomia também recusada. Retorna aos nossos cuidados; feita a enucleação o diagnóstico anatomopatológico revelou melanoma maligno da coróide com invasão pós-laminar do nervo óptico. A importância prognóstica da invasão do nervo óptico por melanoma da coróide ainda não está totalmente esclarecida. Embora raro, tumor pigmentado cobrindo o nervo óptico pode representar melanoma maligno. O diagnóstico diferencial destes casos é geralmente difícil, porém seu reconhecimento à ultra-sonografia ocular é patente e descolamento de retina associado é sinal de atividade tumoral. Os riscos de disseminação da doença exigem atenção na suspeita diagnóstica e conduta precisa.Small-pigmented lesions over the optic disc are very rare and may represent a diagnostic challenge. To report a case of a small malignant choroidal melanoma invading the optic nerve. A 60-year-old male presents with low vision in the left eye due to a small, pigmented lesion over the optic disc. At first the patient refused enucleation. One month later, after further drop in visual acuity, the patient was seen at another service, diagnosed as having a retinal detachment, and pars plana vitrectomy was proposed but also refused by the patient. Returning to our service, the eye was enucleated and a final diagnosis of choroidal melanoma with post-laminar optic nerve invasion was made. Although rare, pigmented lesions over the optic disc may represent a malignant melanoma. The prognostic significance of optic nerve invasion by choroidal melanoma is not clear yet. The differential
Erdoğrul, Özlem Turgay
The presence of Salmonella enteritidis was investigated in 123 liquid whole quail eggs. Salmonella strains were identified and sero-grouped by coagglutination test and slide agglutination test. Seven (5.69%) of 123 whole quail eggs were in group D1 and were sero-typed as Salmonella enteritidis. It was found that in phage-typing of Salmonella enteritidis, three of 7 strains were Salmonella enteritidis PT4 , two of them were PT1, one of them was PT7, and one of them was indefinite.
罗依惠; 陈铭; 李立伟; 钱景; 严杰
objective To determine the existence of virulence-associated invA gene in different genospeeies of Leptospira interrogans reference strains in China.and to understand the alterations of invA gene transcription and expression of L.interrogans strain Lai before or after infecting cells.Methods PCR was applied to detect the invA gene of four L.interrogans strains belonging to four different genospecies and L.biflexa strain Patoc Ⅰ.The entire invA genes from the L.interrogans strains were cloned and then sequenced.The prokaryotic expression system of invA gene of L.interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai was constructed.Using Ni-NTA affinity chromatography,the target recombinant protein rInvA was extracted and purified.Rabbits were immunized with rInvA to obtain antiserum and the titer of antiserum was determined by immunodiffusion test.A model of L interrogans strain Lai infecting human embryo kidney cell line HEK293 was established to detect the alterations of invA gene transcription and expression of the leptospiral strain before or after infecting the host cells by real-time fluorescent quantitative RTPCR and western blot assay.Results All the four tested L.interrogans strains had invA gene whereas L.biflexa strain Patoc Ⅰ not.The similarity of nucleotide and putative amino acid sequences of invA genes from the four L.interrogans strains belonging four different genospecies were 99.33%-100%and 98.66%-100%,respectively.The constructed prokaryotic expression system could efficiently express rInvA and the immunodiffusion titer of rabbit anti-rInvA serum was 1:16.After L.interrogans strain Lai infecting HEK293 cells for 30 min or above,the microbe could adhere the surface of the cells.On the 30 min after the infection,the mRNA level of invA gene of L.interrogans strain Lai was remarkably upregulated,and on the 45 min after infection the mRNA level presented a peak value and then graduated decreased.On the 45 min or 60 min after L
Caffer, M I; Eiguer, T
A significant increase in the number of isolations of Salmonella enteritidis has been observed in Argentina since 1986. Outbreaks of foodborne diseases in humans were associated with the consumption of raw or undercooked hens' eggs. Between 1986 and the first 6 months of 1993 there were 150 outbreaks reported, affecting more than 6000 persons. A total of 71.3% of these outbreaks were confirmed by stool cultures, and 47.3% by bacteriological study of the food implicated in the outbreak. A permanent surveillance of salmonellosis is imperative, taking into account the persistence of Salmonella enteritidis isolations in sporadic cases and in new outbreaks. PMID:8155472
Bispham, J; B.N Tripathi; Watson, P. R.; Wallis, T S
We have used signature-tagged mutagenesis to identify mutants of the host-specific Salmonella enterica serotype Dublin which were avirulent in calves and/or BALB/c mice. A mutant with a transposon insertion in the sseD gene of Salmonella pathogenicity island 2 (SPI-2), which encodes a putative secreted effector protein, was identified. This mutant was recovered from the bovine host but not from the murine host following infection with a pool of serotype Dublin mutants. However, a pure inoculu...
Full Text Available Cationic amino acid transporters (mCAT1 and mCAT2B regulate the arginine availability in macrophages. How in the infected cell a pathogen can alter the arginine metabolism of the host remains to be understood. We reveal here a novel mechanism by which Salmonella exploit mCAT1 and mCAT2B to acquire host arginine towards its own intracellular growth within antigen presenting cells. We demonstrate that Salmonella infected bone marrow derived macrophages and dendritic cells show enhanced arginine uptake and increased expression of mCAT1 and mCAT2B. We show that the mCAT1 transporter is in close proximity to Salmonella containing vacuole (SCV specifically by live intracellular Salmonella in order to access the macrophage cytosolic arginine pool. Further, Lysosome associated membrane protein 1, a marker of SCV, also was found to colocalize with mCAT1 in the Salmonella infected cell. The intra vacuolar Salmonella then acquire the host arginine via its own arginine transporter, ArgT for growth. The argT knockout strain was unable to acquire host arginine and was attenuated in growth in both macrophages and in mice model of infection. Together, these data reveal survival strategies by which virulent Salmonella adapt to the harsh conditions prevailing in the infected host cells.
This thesis reports investigations using gamma-radiation to decontaminate poultry carcasses. The application to foods of doses of ionizing radiation sufficient to reduce the number of viable specific non-sporeforming pathogenic microorganisms so that none is detectable in the treated food by any standard method is termed radicidation. The doses used in this study were at such a level that no undesirable or unfavourable side-effects occurred. The effects of these doses were studied on salmonellae and other microorganisms present in, or associated with poultry carcasses and in liquid and on solid culture media as well. Decimal reduction (D10) values were estimated. These represent the dose (kGy) required to achieve a reduction in initial colony count from N0 to 0.1 N0. Together with the estimation of the numbers of Salmonella present per carcass the data were used to predict the effect of an ionizing radiation treatment of poultry. Data on the effect of ionizing radiation on the total microflora of poultry carcasses were also collected. (Auth.)
Etienne, A; Hecquet, F; Soulard, C; Spinnewyn, B; Clostre, F; Braquet, P
The effects of BN 52021, a new specific paf-acether receptor antagonist and the total Ginkgo Biloba extract (GBE 761) from which this product was isolated, were studied in the rat on paf-acether-induced permeability and cell number changes and on endotoxin-induced lethality. Their activities were compared to those of cyclooxygenase, 5-lipoxygenase and phospholipase A2 inhibitors. BN 52021 given s.c. or orally exerted a dose-related inhibition of paf-acether deleterious effects as well as of endotoxin lethality whereas the other drugs tested were poorly effective. These results strongly suggest paf-acether involvement in endotoxic and septic shock. PMID:3515870
Attachment and colonization of Salmonella enterica serovars to fresh produce leaves was investigated. Biofilm assay and attachment of Salmonella serovars to intact and cut leaves were determined. Salmonella Tennessee and Salmonella Thompson produced stronger biofilms compared to Salmonella Newpor...
Fang, Ferric C.; Weiss, Günter
The hormone hepcidin promotes iron sequestration by macrophages. A recent study by Kim et al (2014) implicates the orphan receptor ERRγ in the regulation of hepcidin production and suggests that targeting the ERRγ-hepcidin axis may be beneficial during infection with the facultative intracellular pathogen Salmonella.
Jonge R de; Hendriks H; Garssen J; Universteit Utrecht, afdeling; MGB; LPI
In human gastrointestinal disease caused by Salmonella, transepithelial migration of neutrophils follows the attachment of bacteria to epithelial tissue. This migration of neutrophils is stimulated by the release of chemokines, including interleukin-8 (Il -8), from the epithelial cells. We have dev
Malaeb, M; Bizri, A R; Ghosn, N; Berry, A; Musharrafieh, U
Salmonellosis is a disease that represents a major public health concern in both developing and developed countries. The aim of this article is to evaluate the public health burden of Salmonella illness in Lebanon. The current scope of the Salmonella infection problem was assessed in relation to disease incidence and distribution with respect to age, gender and district. Factors that provide a better understanding of the magnitude of the problem were explored and highlighted. Data reported to the Epidemiologic Surveillance Department at the Lebanese Ministry of Public Health between 2001 and 2013 was reviewed. Information obtained was compared to information reported regionally and globally. The estimated true incidence was derived using multipliers from the CDC and Jordan. A literature review of all published data from Lebanon about Salmonella susceptibility/resistance patterns and its serious clinical complications was conducted. The estimated incidence was 13·34 cases/100 000 individuals, most cases occurred in the 20-39 years age group with no significant gender variation. Poor and less developed districts of Lebanon had the highest number of cases and the peak incidence was in summer. Reflecting on the projected incidence derived from the use of multipliers indicates a major discrepancy between what is reported and what is estimated. We conclude that data about Salmonella infection in Lebanon and many Middle Eastern and developing countries lack crucial information and are not necessarily representative of the true incidence, prevalence and burden of illness. PMID:26743045
VanCauwenberge, J E; Bothast, R J; Kwolek, W F
Dry heat was used to inactivate Salmonella newington, Salmonella typhimurium, Salmonella anatum, Salmonella kentucky, Salmonella cubana, Salmonella seftenberg, Salmonella thompson, and Salmonella tennessee in corn flour at 10 and 15% moisture. The flour was spray inoculated at 10(5) Salmonella cells per g and then stored at 49 degrees C (120 degrees F); viable Salmonella cells were counted on Trypticase (BBL Microbiology Systems) soy agar plates every 30 min for the first 4 h and then at 4-h ...
Song, Qifa; Shen, Xuanyi; Yang, Yuanbin; Zhang, Danyang; Gao, Hong
Salmonella enterica serotype Enteritidis (S. Enteritidis) is an important causative agent of nontyphoidal salmonellosis in human populations. In this study, we collected 72 S. Enteritidis strains from 2004 to 2014 in Ningbo, mid-east China. Of the 72 strains, we identified a dominant clone of 58 strains recovered from patient's feces (n = 48), blood (n = 1), pleural effusion (n = 1), chickens (n = 3), and dessert cakes (n = 5) by pulsed-field gel electrophoresis (PFGE) and variable-number of tandem repeat analysis (MLVA). The profile arrangements of MLVA were SE1-SE2-SE3-SE5-SE6-SE8-SE9: 4-4-3-11-10-1-3. These dominant strains were susceptible to ampicillin, chloramphenicol, tetracycline, ciprofloxacin, gentamicin, cefotaxime and trimethoprim-sulfamethoxazole, and resistant to nalidixic acid. Additionally, all isolates harboured virulence genes invA, sipA, sopE, and spvB when tested by PCR. Our results reveal that genetically similar S. Enteritidis strains which accounted for several outbreaks as well as blood infection and pleural cavity infection are prevalent in China for a long-term period. This situation calls for further attention in the prevention and control of foodborne disease caused by Salmonella species. PMID:27228342
As part of the Danish RERTR Program, three fuel elements with LEU U3O8-Al fuel and three fuel elements with LEU U3Si2-Al fuel were manufactured by NUKEM for irradiation testing in the DR-3 reactor at the Risoe National Laboratory in Denmark. The specifications for the elements with U3O8-Al fuel are presented here as an illustration only. Specifications for the elements with U3Si2-Al fuel were very similar. In this example, materials, material numbers, documents numbers, and drawing numbers specific to a single fabricator have been deleted. (author)
Braukmann, Maria; Methner, Ulrich; Berndt, Angela
Salmonella serovars are differentially able to infect chickens. The underlying causes are not yet fully understood. Aim of the present study was to elucidate the importance of Salmonella Pathogenicity Island 1 and 2 (SPI-1 and -2) for the virulence of two non-host-specific, but in-vivo differently invasive, Salmonella serovars in conjunction with the immune reaction of the host. Primary avian splenic macrophages were inoculated with Salmonella enterica sub-species enterica serovar (S.) Typhimurium and S. Infantis. The number and viability of intracellular bacteria and transcription of SPI-1 and -2 genes by the pathogens, as well as transcription of immune-related proteins, surface antigen expression and nitric oxide production by the macrophages, were compared at different times post inoculation. After infection, both of the Salmonella serovars were found inside the primary macrophages. Invasion-associated SPI-1 genes were significantly higher transcribed in S. Infantis- than S. Typhimurium-infected macrophages. The macrophages counteracted the S. Infantis and S. Typhimurium infection with elevated mRNA expression of inducible nitric oxide synthase (iNOS), interleukin (IL)-12, IL-18 and lipopolysaccharide-induced tumor necrosis factor alpha factor (LITAF) as well as with an increased synthesis of nitric oxide. Despite these host cell attacks, S. Typhimurium was better able than S. Infantis to survive within the macrophages and transcribed higher rates of the SPI-2 genes spiC, ssaV, sifA, and sseA. The results showed similar immune reactions of primary macrophages after infection with both of the Salmonella strains. The more rapid and stronger transcription of SPI-2-related genes by intracellular S. Typhimurium compared to S. Infantis might be responsible for its better survival in avian primary macrophages. PMID:25811871
Full Text Available Salmonella serovars are differentially able to infect chickens. The underlying causes are not yet fully understood. Aim of the present study was to elucidate the importance of Salmonella Pathogenicity Island 1 and 2 (SPI-1 and -2 for the virulence of two non-host-specific, but in-vivo differently invasive, Salmonella serovars in conjunction with the immune reaction of the host. Primary avian splenic macrophages were inoculated with Salmonella enterica sub-species enterica serovar (S. Typhimurium and S. Infantis. The number and viability of intracellular bacteria and transcription of SPI-1 and -2 genes by the pathogens, as well as transcription of immune-related proteins, surface antigen expression and nitric oxide production by the macrophages, were compared at different times post inoculation. After infection, both of the Salmonella serovars were found inside the primary macrophages. Invasion-associated SPI-1 genes were significantly higher transcribed in S. Infantis- than S. Typhimurium-infected macrophages. The macrophages counteracted the S. Infantis and S. Typhimurium infection with elevated mRNA expression of inducible nitric oxide synthase (iNOS, interleukin (IL-12, IL-18 and lipopolysaccharide-induced tumor necrosis factor alpha factor (LITAF as well as with an increased synthesis of nitric oxide. Despite these host cell attacks, S. Typhimurium was better able than S. Infantis to survive within the macrophages and transcribed higher rates of the SPI-2 genes spiC, ssaV, sifA, and sseA. The results showed similar immune reactions of primary macrophages after infection with both of the Salmonella strains. The more rapid and stronger transcription of SPI-2-related genes by intracellular S. Typhimurium compared to S. Infantis might be responsible for its better survival in avian primary macrophages.
Abstract Salmonellosis is the main cause of human bacterial gastroenteritis in most European countries. Infections with Salmonella is usually subclinical, whereas clinical cases show symptoms with a wide range of severity. Infection is most commonly associated with the consumption of meat, especially poultry or pork, and eggs and their products. Salmonella can enter the food chain at any point throughout its length. The principal reservoir of Salmonellae is the gastrointestinal tract of mamma...
Full Text Available CONTEXT: Salmonella has been identified as a causative agent of acute pancreatitis. OBJECTIVE: We prospectively evaluated the frequency of acute pancreatitis, pancreatic enzyme elevation and morphological pancreatic abnormalities in patients with Salmonella infection. SUBJECTS: Thirty consecutive patients with salmonellosis (Salmonella enterica serovar Enteritidis: n=25; Salmonella enterica serovar Typhimurium: n=5 and 30 sex- and age-matched healthy subjects were studied. MAIN OUTCOME MEASURES: All subjects underwent serum amylase and lipase determination and ultrasonography. RESULTS: None of the subjects developed acute pancreatitis. Two patients (6.7% and two controls showed serum amylase activity above the upper reference limit whereas, in five patients (16.7% and one control subject (3.3%, the serum lipase activity appeared above the upper reference limit. Salmonella infection significantly increased serum activity of lipase (P less than 0.001 while it did not significantly affect serum amylase levels (P=0.204. Serum lipase activity was significantly higher in patients infected by Salmonella enterica serovar Typhimurium than in those infected by Salmonella enterica serovar Enteritidis (P=0.012. Ultrasonography did not show pancreatic abnormalities in any of the subjects. CONCLUSIONS: Our data demonstrated an elevation of serum lipase activity in gastroenteritis due to Salmonella infection, but this elevation does not seem to have clinical significance. The elevation of serum lipase seems to be particularly related to infection from Salmonella enterica serovar Typhimurium.
Hessam A. Halimi
Conclusion: The emergence of multiple antibiotic-resistant strains of Salmonella Typhimurium should be of great concern to the public. No correlation between ERIC fingerprinting and resistance patterns of Salmonella isolates was found, which indicates resistance to antimicrobial agents was not related to specific genetic background.
Arun K. Bhunia
Full Text Available Salmonella enterica is a major food-borne pathogen of world-wide concern. Sensitive and rapid detection methods to assess product safety before retail distribution are highly desirable. Since Salmonella is most commonly associated with poultry products, an evanescent wave fiber-optic assay was developed to detect Salmonella in shell egg and chicken breast and data were compared with a time-resolved fluorescence (TRF assay. Anti-Salmonella polyclonal antibody was immobilized onto the surface of an optical fiber using biotin-avidin interactions to capture Salmonella. Alexa Fluor 647-conjugated antibody (MAb 2F-11 was used as the reporter. Detection occurred when an evanescent wave from a laser (635 nm excited the Alexa Fluor and the fluorescence was measured by a laser-spectrofluorometer at 710 nm. The biosensor was specific for Salmonella and the limit of detection was established to be 103 cfu/mL in pure culture and 104 cfu/mL with egg and chicken breast samples when spiked with 102 cfu/mL after 2–6 h of enrichment. The results indicate that the performance of the fiber-optic sensor is comparable to TRF, and can be completed in less than 8 h, providing an alternative to the current detection methods.
We have developed a rapid and sensitive method for immunomagnetic separation (IMS) of Salmonella along with their real time detection via PCR. Silica-coated magnetic nanoparticles were functionalized with carboxy groups to which anti-Salmonella antibody raised against heat-inactivated whole cells of Salmonella were covalently attached. The immuno-captured target cells were detected in beverages like milk and lemon juice by multiplex PCR and real time PCR with a detection limit of 104 cfu.mL−1 and 103 cfu.mL−1, respectively. We demonstrate that IMS can be used for selective concentration of target bacteria from beverages for subsequent use in PCR detection. PCR also enables differentiation of Salmonella typhi and Salmonella paratyphi A using a set of four specific primers. In addition, IMS—PCR can be used as a screening tool in the food and beverage industry for the detection of Salmonella within 3–4 h which compares favorably to the time of several days that is needed in case of conventional detection based on culture and biochemical methods. (author)
Koyuncu, Sevinc; Andersson, Gunnar; Vos, Pieter; Häggblom, Per
In the present study we investigated if the microarray platforms Premi®Test Salmonella (PTS) and Salmonella array (SA) could be applied for the identification and typing of Salmonella in artificially contaminated animal feed materials. The results were compared to the culture-based MSRV method and serotyping according to Kauffman-White. The SA platform showed a specificity of 100% for the identification of Salmonella compared to 93% with the PTS platform and a sensitivity of 99% or 100%, respectively. Among all identified Salmonella serotypes, 56% with the SA platform and 81% with the PTS platform were correctly identified. The difference in probe signal intensity for each probe was higher between duplicates analyzed with the SA platform than with the PTS platform. Attempts to use the microarray platforms from BPW resulted in many false negative samples and incorrect typing results. The microarray platforms tested were simple to use and might have a potential in tracing studies for Salmonella in the feed chain particularly when rapid information about serotypes are important. PMID:20688409
Jacobsen, Annika; Hendriksen, Rene S.; Aarestrup, Frank Møller;
there is an abundance of accessory genes, including the Salmonella pathogenicity islands (SPIs), transposable elements, phages, and plasmid DNA. We visualize conservation in the genomes in relation to chromosomal location and DNA structural features and find that variation in gene content is localized in a selection......Salmonella enterica is divided into four subspecies containing a large number of different serovars, several of which are important zoonotic pathogens and some show a high degree of host specificity or host preference. We compare 45 sequenced S. enterica genomes that are publicly available (22...... complete and 23 draft genome sequences). Of these, 35 were found to be of sufficiently good quality to allow a detailed analysis, along with two Escherichia coli strains (K-12 substr. DH10B and the avian pathogenic E. coli (APEC O1) strain). All genomes were subjected to standardized gene finding...
Dias, Mariane Rezende; Cavicchioli, Valéria Quintana; Camargo, Anderson Carlos; Lanna, Frederico Germano Piscitelli Alvarenga; Pinto, Paulo Sérgio de Arruda; Bersot, Luciano Dos Santos; Nero, Luís Augusto
Due to the importance of Salmonella spp. in poultry products, this study aimed to track its main contamination routes since slaughtering reception to processing of chicken end cuts. Samples from different steps of slaughtering and processing (n = 277) were collected from two chicken slaughterhouses (Sl1 and Sl2) located in Minas Gerais state, Brazil, and subjected to Salmonella spp. detection. The obtained isolates were subjected to serological identification and tested by PCR for specific Salmonella spp. genes (ompC and sifB). Also, Salmonella spp. isolates were subjected to XbaI macrorestriction and pulsed-field gel electrophoresis (PFGE). Sixty-eight samples were positive for Salmonella spp. and 172 isolates were obtained. Sl1 and Sl2 presented similar frequencies of Salmonella spp. positive samples during reception, slaughtering and processing (p > 0.05), except for higher frequencies in Sl1 for chicken carcasses after de-feathering and evisceration (p < 0.05). PFGE allowed the identification of cross contamination and persistence of Salmonella spp. strains in Sl1. The results highlighted the relevance of the initial steps of chicken slaughtering for Salmonella spp. contamination, and the pre-chilling of carcasses as an important controlling tool. In addition, the presence of Salmonella spp. in chicken end cuts samples represents a public health concern. PMID:27162388
Adson Santa Cruz Oliveira
Full Text Available
The invasive capacity and persistence of this pathogen, crop and ceca in apparently healthy birds of two broiler lines raised without growth promoter antibiotics in ration and originated from eggs inoculated eggshell and in allantoidal cavity with Salmonella Enteritidis. Histological and bacteriological exams from cecal and crop were performed with one, seven, 14 and 21 days of age after hatch in broilers of fast and slow growing rate. Bacterio-logical exams were performed fecal excretion with one, eigth, 22 and 35 days. The Salmonella Enteritidis invaded and colonizated the gastrointestinal tract of the two lines tested, but the the infection reduced with age, and was more persistant in Ross broilers. The results were different for two lines. The pathogen was excreted from just one chick of ISA Label at 22 days of age and four Ross chicks until 35 days of age. In order, Salmonella was detected in 87.5% (14/16 and 38,1% (5/16 of ceca; in 81.2% (13/16 and 12.5% (2/16 of crops; in fast and slow growing rate lines, respectively. In apparent healthy organs, excepted the crop, an inflammatory process with predominance of macrophage and lymphocytes. The slow growing rate line was effective to eliminate bacteria in the organism.
Key-words: Ceca, crop, fecal excretion, inflammation.
Avaliaram-se, neste estudo, a capacidade inva-siva, a persistência e a freqüência de excreção fecal da Salmonella Enteritidis em aves aparentemente saudáveis de duas linhagens de frango de corte, criadas sem antibióticos promotores de crescimento na ração e oriundas de ovos inoculados na casca ou na cavidade alantóide com Salmonella Enteritidis fagotipo 4. Realizaram-se exames bacteriológicos das excretas com um, oito, 22 e 35 dias, e histológicos e bacteriológicos do inglúvio e ceco, com um, sete, quatorze e 21 dias pós-eclosão em frangos de crescimento rápido e lento. Salmonella
Hernandez, Jorge; Lindberg, Peter; Waldenström, Jonas; Drobni, Mirva; Olsen, Björn
A novel Salmonella serovar was isolated from Peregrine falcon (Falco peregrinus) nestlings in northern Sweden in 2006. Three isolates of the same clone was retrieved from three falcon siblings and characterized as Salmonella enterica sub-species enterica: O-phase 13, 23:-: e, n, z 15 and the H-phase was not present. We propose the geographical name Salmonella enterica, sub-species entericaserovar Pajala to this novel Salmonella.Keywords: Salmonella; epidemiology; ecology; peregrine falcon; no...
吴晓芳; 韩建康; 纪蕾; 徐德顺; 陈莉萍; 沈月华; 查赘峰
目的 建立可同时检测沙门菌、单增李斯特菌的双重实时荧光PCR快速检测方法,并应用于食品样品的检测.方法 根据GenBank上下载的沙门菌invA基因、单增李斯特菌hlyA基因的保守区序列分别设计特异性引物和TaqMan探针,建立并优化双重实时荧光PCR反应体系.对该方法的特异性、敏感性和稳定性进行评估,并应用于食品标本的检测.结果 实验结果表明,该检测方法特异性强,对大肠埃希菌、志贺菌、副溶血性弧菌等其他病原菌进行检测均未产生交叉反应.对沙门菌和单增李斯特菌纯培养物的最低检出限分别可达10 cfu/ml和100cfu/ml;并且重复性好,变异系数均小于5%;对80份食品标本同时采用本文建立的双重荧光PCR方法和单重荧光PCR方法以及传统的分离培养方法进行检测,结果显示本文建立的双重荧光PCR方法对两种病原菌的检测效果明显优于单重荧光PCR方法以及传统的分离培养法,整个检测过程可在10 h内完成,包括前增菌所用的6 h.结论 本研究建立的多重实时荧光PCR方法能同时对沙门菌和单增李斯特菌进行快速检测,并且灵敏度高、特异性好,可为食源性疾病的病原学快速检测提供新的手段.%Objective To establish a TaqMan-based multiplex real-time PCR assay for the detections of Salmonella and Listeria monocytogenes and conduct Salmonella and Listeria monocytogenes detections in food samples. Methods The specific primers and probes were designed in the conserved region of the invA gene for Salmonella and in the hlyA gene for Listeria monocytogenes, respectively. The reaction conditions were optimized, and the sensitivity, specificity and the stability of the assay were evaluated. The food samples collected from the supermarket were detected by this assay. Results The results showed that the assay had high specificity for the detections of Salmonella and Listeria monocytogenes without any
Herrero-Fresno, Ana; Rodicio, Rosaura; Montero, Ignacio; García, Patricia; Rodicio, M Rosario
Five variants of a resistant derivative of pSLT (termed pUO-StVR2) were detected in clinical isolates of Salmonella enterica serovar Typhimurium recovered in Spain. The structure of these variants revealed the involvement of IS1, IS26 and Tn21-like transposition, as well as homologous recombinati...... understand the co-selection mechanisms that are helping to maintain multidrug resistance in bacterial pathogens, despite the structural instability of the responsible DNA....
N. O. Facure
Full Text Available Registro de caso de paciente com infiltração leptomeníngea por metás-tase de mieloma múltiplo. A infiltração neoplásica ocorria nas leptomeninges da base do encéfalo e, em especial, nas do canal raqueano. O tecido tumoral formava manguito no espaço sub-aracnóideo que envolvia os segmentos cervical e torácico da medula espinhal. A partir dos segmentos lombares a medula se achava infiltrada pelo tumor; as raízes da cauda equina achavam-se também invadidas pelo tecido neoplásico. Não foi feito o diagnóstico em vida. O quadro clínico caracterizava-se por sinais de irritação meníngea e de sofrimento radículo-medular a partir dos segmentos lombares altos e o quadro liquórico, por reação inflamatória de tipo sub-agudo que determinava bloqueio do canal raqueano. Os autores chamam a atenção para a raridade da invasão leptomeníngea por mieloma múltiplo e para a dificuldade diagnostica do caso. Neste último sentido discutem os dados do quadro liquórico bem como salientam não ter sido possível completar o estudo histoquímico das células plasmocitárias.A case of a patient with multiple myelomatosis that presented diffuse leptomeningeal involvement by metastatic tissue is reported. The diagnosis was based upon necroscopic examination. The leptomeninges were infiltrated by neoplastic cells, the infiltration being more evident in the leptomeninges of the spinal canal. In this region the subarachnoid space was filled by neoplastic tissue that surrounded the spinal cord and cauda equina. The lumbal and sacral portions of spinal cord were invaded by tumor cells. Fever, congestion of left eye and signs of leptomeningeal irritation appeared at first and were followed by crural paraplegia about two months later. At this occasion a CSF examination showed changes proper to subacute inflammatory process and the manometric test of Stookey suggested the occurence of blockage in the spinal canal. The unusual leptomeningeal involvement
Melo, Airis Maria Araújo; Alexandre, Dalila L; Furtado, Roselayne F; Borges, Maria F; Figueiredo, Evânia Altina T; Biswas, Atanu; Cheng, Huai N; Alves, Carlúcio R
Pathogen detection is a critical point for the identification and the prevention of problems related to food safety. Failures at detecting contaminations in food may cause outbreaks with drastic consequences to public health. In spite of the real need for obtaining analytical results in the shortest time possible, conventional methods may take several days to produce a diagnosis. Salmonella spp. is the major cause of foodborne diseases worldwide and its absence is a requirement of the health authorities. Biosensors are bioelectronic devices, comprising bioreceptor molecules and transducer elements, able to detect analytes (chemical and/or biological species) rapidly and quantitatively. Electrochemical immunosensors use antibody molecules as bioreceptors and an electrochemical transducer. These devices have been widely used for pathogen detection at low cost. There are four main techniques for electrochemical immunosensors: amperometric, impedimetric, conductometric, and potentiometric. Almost all types of immunosensors are applicable to Salmonella detection. This article reviews the developments and the applications of electrochemical immunosensors for Salmonella detection, particularly the advantages of each specific technique. Immunosensors serve as exciting alternatives to conventional methods, allowing "real-time" and multiple analyses that are essential characteristics for pathogen detection and much desired in health and safety control in the food industry. PMID:27138197
Irfan; Ahmad; Mir; Sudhir; Kumar; Kashyap; Sunil; Maherchandani
Objective: To study the occurrence and serotype diversity of Salmonella isolates in different species of poultry(chicken, emu and duck) and determine their resistance pattern against various antibiotics of different classes.Methods: About 507 samples comprising 202 caecal contents and 305 fecal samples from chicken, emu and duck were processed for isolation of Salmonella enterica. Salmonellae were isolated and detected by standard protocol of ISO 6579 Amendment 1: Annex D. Genetic confirmation was also made by using 16 S r RNA genus specific PCR. Serotype specific PCR was also done to detect the most common serovars viz. Salmonella Enteritidis, Salmonella Typhimurium and Salmonella Gallinarum. All obtained isolates were subjected to a set of 25 antibiotics to study their antibiogram by using Baeur-Kirby disk diffusion method.Results: Out of 507 samples processed, 32 isolates of Salmonella enterica(18 from caecal contents and 14 from faecal samples) were obtained, of which 24 belonged to 6 different serovars, 6 were untypeable and 2 were rough strains. Salmonella Enteritidis was the most predominant serotype(9), followed by Salmonella Typhimurium(5), Salmonella Virchow(4), Salmonella Gallinarum(3), Salmonella Reading(2) and Salmonella Altona(1). Antibiotic resistance pattern was maximum(100%) to oxacillin, penicillin and clindamycin, followed by ampicillin(68.75%), tetracycline(65.62%), nalidixic acid(56.25%) and colistin(46.87%). High sensitivity of isolates was recorded for chloramphenicol(96.87%) followed by meropenem(84.37%). Conclusions: Occurrence of high proportion of serovars in our study which can cause serious gastroenteritis in humans is a matter of concern. Salmonella Altona has been detected for the first time in India from poultry. This serotype is known to cause serious outbreaks of gastroenteritis in humans. Multidrug resistant isolates were recovered at high percentage which can be attributed to non-judicious use of antibiotics both in prophylaxis
Irfan Ahmad Mir; Sudhir Kumar Kashyap; Sunil Maherchandani
Objective:To study the occurrence and serotype diversity of Salmonella isolates in different species of poultry (chicken, emu and duck) and determine their resistance pattern against various antibiotics of different classes. Methods:About 507 samples comprising 202 caecal contents and 305 fecal samples from chicken, emu and duck were processed for isolation of Salmonella enterica. Salmonellae were isolated and detected by standard protocol of ISO 6579 Amendment 1:Annex D. Genetic confirmation was also made by using 16S rRNA genus specific PCR. Serotype specific PCR was also done to detect the most common serovars viz. Salmonella Enteritidis, Salmonella Typhimurium and Salmonella Gallinarum. All obtained isolates were subjected to a set of 25 antibiotics to study their antibiogram by using Baeur-Kirby disk diffusion method. Results:Out of 507 samples processed, 32 isolates of Salmonella enterica (18 from caecal contents and 14 from faecal samples) were obtained, of which 24 belonged to 6 different serovars, 6 were untypeable and 2 were rough strains. Salmonella Enteritidis was the most predominant serotype (9), followed by Salmonella Typhimurium (5), Salmonella Virchow (4), Salmonella Gallinarum (3), Salmonella Reading (2) and Salmonella Altona (1). Antibiotic resistance pattern was maximum (100%) to oxacillin, penicillin and clindamycin, followed by ampicillin (68.75%), tetracycline (65.62%), nalidixic acid (56.25%) and colistin (46.87%). High sensitivity of isolates was recorded for chloramphenicol (96.87%) followed by meropenem (84.37%). Conclusions:Occurrence of high proportion of serovars in our study which can cause serious gastroenteritis in humans is a matter of concern. Salmonella Altona has been detected for the first time in India from poultry. This serotype is known to cause serious outbreaks of gastroenteritis in humans. Multidrug resistant isolates were recovered at high percentage which can be attributed to non-judicious use of antibiotics both in
Soltan Dallal MM; Khalilian M; Masoumi Asl H; Bakhtiari R; Davoodabadi A; Rajabi Z
Background and aim: Foodborne diseases are a major problem of modern society. The epidemiological investigations in many parts of the world had been shown that infections caused by Salmonella serovars are increasing. Therefore, the identification of Salmonella species to serovars level is in specific importance. In addition, the emergence of drug resistance among these serovars also has become a major global concern. So, the knowledge of the resistance patterns of Salmonella could be very use...
Bergman, Molly A.; Cummings, Lisa A.; Barrett, Sara L. Rassoulian; Smith, Kelly D.; Lara, J. Cano; Aderem, Alan; Cookson, Brad T.
A better understanding of immunity to infection is revealed from the characteristics of microbial ligands recognized by host immune responses. Murine infection with the intracellular bacterium Salmonella generates CD4+ T cells that specifically recognize Salmonella proteins expressed in bacterial surface organelles such as flagella and membrane vesicles. These natural Salmonella antigens are also ligands for Toll-like receptors (TLRs) or avidly associated with TLR ligands such as lipopolysacc...
Salmonellas are the main responsible agent for the frequent food-borne gastrointestinal diseases. Their detection using classical methods are laborious and their results take a lot of time to be revealed. In this context, we tried to set up a revealing technique of the invA virulence gene, found in the majority of Salmonella species. After amplification with PCR using specific primers created and verified by bioinformatics programs, two couples of primers were set up and they appeared to be very specific and sensitive for the detection of invA gene. (Author)
Christensen, H.; Nordentoft, Steen; Olsen, J.E.
To establish the phylogenetic relationships between the subspecies of Salmonella enterica (official name Salmonella choleraesuis), Salmonella bongori and related members of Enterobacteriaceae, sequence comparison of rRNA was performed by maximum-likelihood analysis. The two Salmonella species were...
Wegener, Henrik Caspar; Hald, Tine; Wong, Danilo Lo Fo; Madsen, M.; Korsgaard, Helle; Bager, Flemming; Gerner-Smidt, P.; Mølbak, K.
We describe Salmonella control programs of broiler chickens, layer hens, and pigs in Denmark. Major reductions in the incidence of foodborne human salmonellosis have occurred by integrated control of farms and food processing plants. Disease control has been achieved by monitoring the herds and...... flocks, eliminating infected animals, and diversifying animals (animals and products are processed differently depending on Salmonella status) and animal food products according to the determined risk. In 2001, the Danish society saved U.S.$25.5 million by controlling Salmonella. The total annual...
Hald, Tine; Lund, Jan
sources. The interface is based on a Salmonella source-attribution model developed for setting target for Salmonella in the turkey production: the Turkey Target Source Attribution Model (TT-SAM). Results from this model were used by the BIOHAZ panel in their related Scientific Opinion. The developed......Project developed on the procurement project NP/EFSA/BIOHAZ/2011/04 EFSA has been working on a series of Scientific Opinions originating from a mandate received by the European Commission (EC) in July 2008 on the review of Salmonella targets in poultry primary production. For evaluating targets in...... the broiler and turkey production, specific Salmonella source attribution models have been developed by external contractors. Both models were based on the Hald model and use a Bayesian approach employing microbial subtyping data, in both cases Salmonella serovar data. These types of source...
Chen, Yu; Wu, Da; Sun, Min; Deng, Mingjun; Cui, Shuhua; Liang, Chengzhu; Geng, Juan; Sun, Tao; Long, Ling; Xiao, Xizhi
While inspecting animal feed for Salmonella contamination, we routinely observed bacterial colonies on selective agars that were similar in appearance to those formed by Salmonella. These were identified as Citrobacter freundii, Proteus mirabilis, and Serratia fonticola using biochemical and serological techniques. Because the presence of these bacterial species confounds identification of Salmonella, we refer to them as "interference bacteria." Polyvalent antisera against these interference bacteria were prepared by immunizing rabbits with a mixture of all three organisms. To minimize or eliminate interference by these bacteria, the polyvalent antisera were introduced between the steps of selective enrichment and Salmonella-selective plating. The antisera raised against the interference bacteria, when combined with neonatal rabbit complement, exhibited specific bactericidal activity against C. freundii, P. mirabilis, and S. fonticola. The respective serum bactericidal assay titers were 2(9), 2(8), and 2(10). In selective broth, polyvalent antisera could also kill the target bacterial cells effectively. We tested 526 samples (186 white fishmeal, 97 red fishmeal, and 243 cattle bone powder) using the polyvalent antisera and found that the rates of contamination of each species of the three respective foods decreased by 58.8, 100, and 83%. Our data indicates that polyvalent sera against C. freundii, P. mirabilis, and S. fonticola can be used as inhibitors to increase the accuracy of Salmonella detection. PMID:26822285
Poltronieri, Palmiro; Cimaglia, Fabio; De Lorenzis, Enrico; Chiesa, Maurizio; Mezzolla, Valeria; Reca, Ida Barbara
Food pathogens are the cause of foodborne epidemics, therefore there is a need to detect the pathogens in food productions rapidly. A pre-enrichment culture followed by selective agar plating are standard detection methods. Molecular methods such as qPCR have provided a first rapid protocol for detection of pathogens within 24 h of enrichment culture. Biosensors also may provide a rapid tool to individuate a source of Salmonella contamination at early times of pre-enrichment culture. Forty mL of Salmonella spp. enrichment culture were processed by immunoseparation using the Pathatrix, as in AFNOR validated qPCR protocols. The Salmonella biosensor combined with immunoseparation showed a limit of detection of 100 bacteria/40 mL, with a 400 fold increase to previous results. qPCR analysis requires processing of bead-bound bacteria with lysis buffer and DNA clean up, with a limit of detection of 2 cfu/50 μL. Finally, a protein chip was developed and tested in screening and identification of 5 common pathogen species, Salmonella spp., E. coli, S. aureus, Campylobacter spp. and Listeria spp. The protein chip, with high specificity in species identification, is proposed to be integrated into a Lab-on-Chip system, for rapid and reproducible screening of Salmonella spp. and other pathogen species contaminating food productions. PMID:27110786
Full Text Available In 1995 several outbreaks of food poisoning in humans occurred in Iceland, that were traced to salmonella contamination of singed sheep heads. This prompted us to study the prevalence of salmonella infection in sheep and to trace where and how infection might have occurred. Faecal, intestinal contents and tonsillar samples were collected in the spring and autumn from sheep on 50 farms in the southwestern part of the country, where salmonellosis had been detected and from 5 farms in the northwestern part of the country. All faecal samples from the southwest were negative, whereas samples from 3 farms obtained in the autumn in the northwest were positive. Tonsillae taken in the autumn were positive in sheep from 3 farms in the southwest and 2 in the northwest. Our results show that salmonella infection is rare in Icelandic sheep but healthy carriers may harbour the bacteria in tonsillae. Salmonella was not detected in drainage from slaughterhouses nor in singed sheep heads.
Wild, Philipp; Farhan, Hesso; McEwan, David G; Wagner, Sebastian; Rogov, Vladimir V; Brady, Nathan R; Richter, Benjamin; Korac, Jelena; Waidmann, Oliver; Choudhary, Chunaram; Dötsch, Volker; Bumann, Dirk; Dikic, Ivan
Selective autophagy can be mediated via receptor molecules that link specific cargoes to the autophagosomal membranes decorated by ubiquitin-like microtubule-associated protein light chain 3 (LC3) modifiers. Although several autophagy receptors have been identified, little is known about mechanisms...... controlling their functions in vivo. In this work, we found that phosphorylation of an autophagy receptor, optineurin, promoted selective autophagy of ubiquitin-coated cytosolic Salmonella enterica. The protein kinase TANK binding kinase 1 (TBK1) phosphorylated optineurin on serine-177, enhancing LC3 binding...... affinity and autophagic clearance of cytosolic Salmonella. Conversely, ubiquitin- or LC3-binding optineurin mutants and silencing of optineurin or TBK1 impaired Salmonella autophagy, resulting in increased intracellular bacterial proliferation. We propose that phosphorylation of autophagy receptors might...
... recommendations as specified under 40 CFR 79.60, the following specific information shall be reported: (i... consulted. (1) 40 CFR 798.5265, The Salmonella typhimurium reverse mutation asay. (2) Ames, B.N., McCann, J... mutation assay. 79.68 Section 79.68 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY...
Recent work has called attention to the unequal competitive abilities of different Salmonella serotypes in standard broth culture and plating media. Such serotypes include Enteritidis and Typhimurium that are specifically targeted in some regulatory and certification programs because they cause a l...
Full Text Available Salmonella enterica, the most pathogenic species of the genusSalmonella, includes more than 2,500 serovars, many of which are of great veterinary and medical significance. The emergence of food-borne pathogens, such as Salmonella spp., has increased knowledge about the mechanisms helping microorganisms to persist and spread within new host populations. It has also increased information about the properties they acquire for adapting in the biological environment of a new host. Thedifferences observed between serovars in their host preference and clinical manifestations are referred to as “serovar-host specificity” or “serovar-host adaptation”. The genus Salmonella, highly adaptive to vertebrate hosts, has many pathogenic serovars showing host specificity. Serovar Salmonella Typhi, causing disease to man and higher primates, is a good example of host specificity. Thus, understanding the mechanisms that Salmonella serovars use to overcome animal species' barriers or adapt to new hosts is also important for understanding the origins of any other infectious diseases or the emergence of new pathogens. In addition, molecular methods used to study the virulence determinants of Salmonella serovars, could also be used to model ways of studying the virulence determinants used by bacteria in general, when causing disease to a specific animal species
Alicia Velázquez Pérez
Full Text Available Se presenta el caso de una paciente femenina de color blanco y dos años de edad, con diagnóstico prenatal de sicklemia, que desde edades tempranas tiene problemas de la enfermedad. Ingresó en esta ocasión por una de las complicaciones infecciosas que ocasiona este padecimiento, una osteomielitis del húmero izquierdo, aislándose el germen en el hemocultivo realizado, una salmonella. Necesitó de tratamiento enérgico y prolongado; se obtuvo un resultado satisfactorio en la evolución de la enfermedad y se sigue sistemáticamente por consulta externa en la actualidad
Hessam A Halimi; Hesam A Seifi; Mehrnaz Rad
Objective:To evaluate serovar and antimicrobial resistance patterns of Salmonella spp isolated from healthy, diseased and necropsied cows and calves in this observational study. Methods:Nineteen isolates recovered from feces and tissues of salmonellosis-affected animals of two commercial farms in north-east of Iran. In second part of the study, the two farms were sampled 4 times with an interval of 2 month. The samples included calves’ feces, adult cows’ feces, feeds, water, milk filters, and milk fed to calves. Five Salmonella were isolated from 332 fecal samples collected from calves and peri-parturient cows. No Salmonella was recovered from water, feed, milk filers and milk fed to calves. Results:Salmonella Typhimurium was the most frequently isolate among all sero-groups. S. Dublin was only accounted for 8%(two out of 24) of isolates. Isolated Salmonella strains were used for the ERIC PCR DNA fingerprinting assay. Our results grouped Salmonella isolates into 3 clusters, suggesting that specific genotypes were responsible for each sero-group of Salmonella. The results also revealed diversity among Salmonella isolates in cluster III (sero-group B). Eighteen out of 19 Salmonella spp. were resistant to oxytetracycline. Five isolates out of 19 showed more than one drug resistance. Multi-drug resistance was seen only among Salmonella Typhimurium isolates. Enrofloxacin was the most susceptible antibiotic against all isolates in this study. Conclusion:The emergence of multiple antibiotic-resistant strains of Salmonella Typhimurium should be of great concern to the public. No correlation between ERIC fingerprinting and resistance patterns of Salmonella isolates was found, which indicates resistance to antimicrobial agents was not related to specific genetic background.
Full Text Available A novel Salmonella serovar was isolated from Peregrine falcon (Falco peregrinus nestlings in northern Sweden in 2006. Three isolates of the same clone was retrieved from three falcon siblings and characterized as Salmonella enterica sub-species enterica: O-phase 13, 23:-: e, n, z 15 and the H-phase was not present. We propose the geographical name Salmonella enterica, sub-species enterica serovar Pajala to this novel Salmonella.
Full Text Available Plant essential oils are natural products extracted from plants and because of their antimicrobial properties can be used as natural additives in foods. They are also useful for decontamination of food-borne pathogens and can be a safe additive in foods. The antimicrobial activities of essential oils belonging to Saturiea hortensis, Thymus vulgaris, Mentha polegium, Cuminum cyminum, Lavandula officinalis and Mentha viridis L. (spearmint were investigated at different concentrations (0.1, 0.3, 0.5, 1, 2, 5 and 10%v/v against Salmonella typhimurium, Salmonella paratyphi A and Salmonella paratyphi B by using the agar well diffusion method. Essential oils showed inhibitory effect on Salmonella spp. in the agar well diffusion assay. In addition, the capability of essential oils for decontamination of minced row beef, ground beef, minced raw chicken and minced raw fish inoculated with Salmonella spp. at 0.1 and 0.5%v/v were assessed. Reduction of the Salmonella spp. population was observed following the inoculation of the cultures with 0.1 and 0.5%v/v essential oils.
Acute diarrhea associated with Salmonella enterica in Belo Horizonte-MG: prevalence and characterization of isolates Diarreia aguda associada a Salmonella enterica em Belo Horizonte-MG: prevalência e caracterização das amostras isoladas
Mireille Ângela Bernardes Sousa
Full Text Available Introduction: Acute infectious diarrhea is still regarded as a public health problem associated with a wide range of etiologic agents, from which Salmonella enterica is particularly worth mentioning inasmuch as it is a major cause of inflammatory diarrhea in both developed and developing countries. Objective: To assess the distribution of S. enterica among children with acute diarrhea in Belo Horizonte and to characterize bacterium isolates. Material and methods: The study group comprised a total of 157 children from low socioeconomic background. Stool samples were collected for leukocyte analysis and Salmonella bacterial culture. The isolates were serotyped and evaluated as to antimicrobial susceptibility profile, extended-spectrum β-lactamases (ESBL production, and presence of virulence markers (invA, iroB, and spvC. RESULTS: A total of 5/3.2% children were infected by S. enterica, 3/60% by S. enterica Typhimurium, 1/20% by S. enterica Enteritidis and 1/20% S. enterica subsp. enterica serotype 8.20:z4,z23:-. Fecal leucocytes were detected in two out of five fecal specimens positive for S. enterica. Isolates from three children were resistant to nalidixic acid, nalidixic acid + chloramphenicol, and nalidixic acid + chloramphenicol + ampicillin. ESBL production was not detected. All samples presented invA and iroB genes. spvC marker was observed in isolates from two children infected by S. Typhimurium and S. Enteritidis. Conclusion: The results demonstrate that S. enterica infection is uncommon among children from our region. Furthermore, they indicate the need for periodic monitoring of bacterial antimicrobial susceptibility profile in order to establish suitable antimicrobial therapy when required.INTRODUÇÃO: A diarreia infecciosa aguda é considerada um problema de saúde pública associado a uma ampla gama de agentes etiológicos, entre os quais destaca-se Salmonella enterica, causa importante de diarreia inflamatória em pa
Bird, C B; Miller, R L; Miller, B M
The Reveal for Salmonella (RSS) test system is a presumptive qualitative test that detects the presence of Salmonella organisms in foods within 21 h total testing time, allowing the user to release negative products 24 h earlier than when using other rapid test kits. Foods are enriched with a proprietary resuscitation medium called Revive and then selectively enriched with either Selenite Cystine or Rappaport-Vassiliadis selective media. The enriched culture is used to inoculate the RSS detection device, which initiates a lateral flow through a reagent zone containing anti-Salmonella antibodies conjugated to colloidal gold particles that capture antigens present in the culture. The antigen-antibody complex migrates farther and is captured by an additional anti-Salmonella antibody, causing the colloidal gold to precipitate and form a visual line, indicating a positive result. A procedural control line also will form regardless of the presence of Salmonella organisms to indicate the test is working properly. Existing AOAC Official Methods for Salmonella organisms require a 48 h enrichment before testing. Hence, a food product has to be held before release, adding extra cost to the company and the consumer. The RSS test system was evaluated by quantitative spiking studies. Although AOAC encourages inclusion of naturally contaminated foods, almost all microbiological AOAC validation studies have been performed with artificially contaminated foods for absolute control over the study. The RSS test system is designed to test many food types for Salmonella organisms and has a limit of detection of 5-10 colony-forming units (cfu)/25 g with a false-negative rate of < 1% and a false-positive rate of < 5.0%. It showed an 81% overall agreement with the traditional procedure of the U.S. Department of Agriculture's Food Safety Inspection Service. PMID:10367381
Fahimeh Baghbani-arani; Mercedeh Tajbakhsh; Atiyeh Hashemi Soltaniyeh; Bahareh Rajaei; Seyed Davar Siadat; Mohamahreza Aghasadeghi; Seyed Mehdi Sadat
Background & Objective: Molecular typing is an important tool in surveillance and outbreak investigations of human Salmonella infections. In this study, Subtyping of Salmonella Paratyphi B and C isolates derived from Iranian patients was carried out by RAPD-PCR to assess the extent of genetic diversity of these isolates. Materials & Methods: Fourteen Salmonella isolates including 6 strains of Salmonella paratyphi B and 8 strains of Salmonella paratyphi C were characterized using RAPD-PCR. Two...
The O antigen polysaccharide of Salmonella montevideo was isolated from a core-defective mutant by the phenol/water procedure, and was suspected to contain phosphomonester and cyclic phosphodiester at its reducing end in anology to the O hapten from Salmonella typhimurium (Kent and Obsborn, 1968. Therefore, it was chromatographed on a DEAE-cellulose column. Whereas one part eluted with water the other part of the polysaccharide could only be eluted with buffer. Both fractions were further purified on Sephadex G100 and contained mannose, glucose, N-acetylglucosamine and phosphate in a molar ratio of 4:1:1: less than 0.1. In order to specifically label the reducing end phosphate was removed enzymatically, or the presumed cyclic diester was cleaved by mild hydrolysis, and the fractions were reduced with sodium horo[3H]hydride. Both fractions yield mainly [3H]glycerol after hydrolysis and paper chromatogaphy. In addition, [3H]mannitol and [H]monohydroxyacetone could be identified by paper chromatography and were concluded to be the result of phosphate migration and beta-elimination reactions taking place during the isolation procedure and the various treatments prior to sodium boro[3H]hydride reduction. These findings in addition to periodate oxidation studies indicated that the O antigen polysaccharide of Salmonella montevideo had glyceraldehyde phosphate at its reducing end. From the incorporation of 3H into the polysaccharide the O antigen was calculated to consist of about 19 repeating units of 6 sugar residues each. PMID:1097244
Manning, Johanna; Gole, Vaibhav; Chousalkar, Kapil
Salmonellosis is a significant zoonotic disease which has a considerable economic impact on the egg layer industry. There is limited information about the prevalence of Salmonella spp. in backyard chickens. The current study was conducted to determine the prevalence of Salmonella in backyard chickens, and the associated virulence of any serovars identified. Hundred and fifteen pooled samples from 30 backyard flocks in South Australia were screened. Four flocks tested positive for Salmonella spp. The overall Salmonella isolation rate in the current study was 10.4%. The estimated prevalence at individual bird level was 0.02% (95% CI 0.025-0.975). The serovars isolated were Salmonella Agona, Salmonella subsp 2 ser 21:z10:z6 (Wandsbek) and Salmonella Bovismorbificans. All Salmonella isolates tested positive for the prgH, orfL and spiC genes. The Salmonella subsp 2 ser 21:z10:z6 (Wandsbek) had the most antibiotic resistance, being resistant to ampicillin and cephalothin and having intermediate resistance to florphenicol. All of the Salmonella Agona had intermediate resistance to the ampicillin, while the Salmonella Bovismorbificans were susceptible to all antibiotics tested. With the increased interest of keeping backyard chickens, the current study highlights the zoonotic risk from Salmonella spp. associated with home flocks. PMID:25899620
ROBERTO GOMES NOGUEIRA; VANESSA GRANATO TOSCANO; NITAMAR ABDALA; JOÃO NORBERTO STÁVALE; Suzana M. Fleury Malheiros; ALBERTO ALAIN GABBAI
Descrevemos o caso de um paciente com diagnóstico de germinoma nos núcleos da base e tálamo, com invasão do tronco cerebral, ressaltando as características observadas nos exames de tomografia computadorizada e de ressonância magnética, os possíveis diagnósticos diferenciais, e a necessidade da comprovação anátomo-patológica, por tratar-se de localização pouco frequente de um tumor com possibilidade de evolução favorável após tratamento com quimio e radioterapia.We report an unusual case of ge...
Full Text Available A total of 400 hatchery samples comprised of yolk interior (100, paper pad (100, shell membrane (100 and fecal swab of newly hatched chicks (100 were tested to detect the presence of Salmonella organism by bacteriological agar plate test. Positive cases recorded in this study were 37 (37, 12 (12, 3 (3 and 19 (19% from each sample (100 of yolk interior, paper pad, shell membrane and fecal swab of newly hatched chicks, respectively. A representative numbers of 50 isolates were used for the identification of serogoups of Salmonella prevailing in selected area by using polyvalent antisera. The result indicated that the test isolates 45(90% were typed to a specific serogroup of "O". All 45 isolated Salmonella serogroup ?O? were then characterized by different specific biochemical media. Based on these tests, the selective isolates were identified as Salmonella gallinarum.
The sums propose to evaluate the bacterial contamination of certain food taken randomly by two pathogenic bacteria (Salmonella and Listeria) considering the evolution of the diseases of food oignon. For that 78 food samples of different origins were analysed. 2 stocks of the Listeria kind and 3 stocks of the salmonella kind were insulated and identified by biochemical and molecular tests. The pathogenic isolates were identified by coloration gram, test catalase, insulation on specific culture media and Api (20 E for Salmonella and Api listeria. At the end, the PCR were realized to amplify the gene iap which codes for the protein p60 at listeria as well as a sequence clonee randomly specific of Salmonella.
EFSA Panel on Biological Hazards (BIOHAZ); Scientific Opinion on a review on the European Union Summary reports on trends and sources zoonoses, zoonotic agents and food-borne outbreaks in 2009 and 2010 – specifically for the data on Salmonella, Campylobacter, verotoxigenic Escherichia coli
insight. Ultimately, summary measures of public health such as disability adjusted life years (DALYs) and cost-of-illness estimates should be presented. Travel information was found to be still incomplete in many MSs. For many pathogens this hampers source attribution. To better understand the public...... health problems related to food and animal sources in the EU, it is desirable to differentiate between travel within and outside the EU. This would also be useful to better evaluate the public health impact of EU-wide food safety measures. Whenever possible the data/results should be analysed using......The European Union (EU) Summary Reports on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2009 and 2010 – specifically for the data on Salmonella, Campylobacter, verotoxigenic Escherichia coli, Listeria monocytogenes and foodborne outbreaks was reviewed. The main...
Berge, A C; Wierup, M
Nutritional strategies to minimize Salmonella in food animal production are one of the key components in producing safer food. The current European approach is to use a farm-to-fork strategy, where each sector must implement measures to minimize and reduce Salmonella contamination. In the pre-harvest phase, this means that all available tools need to be used such as implementation of biosecurity measures, control of Salmonella infections in animals at the farm as well as in transport and trade, optimal housing and management including cleaning, disinfection procedures as well as efforts to achieve Salmonella-free feed production. This paper describes some nutritional strategies that could be used in farm control programmes in the major mono-gastric food production animals: poultry and pigs. Initially, it is important to prevent the introduction of Salmonella onto the farm through Salmonella-contaminated feed and this risk is reduced through heat treatment and the use of organic acids and their salts and formaldehyde. Microbiological sampling and monitoring for Salmonella in the feed mills is required to minimize the introduction of Salmonella via feed onto the farm. In addition, feed withdrawal may create a stressful situation in animals, resulting in an increase in Salmonella shedding. Physical feed characteristics such as coarse-ground meal to pigs can delay gastric emptying, thereby increasing the acidity of the gut and thus reducing the possible prevalence of Salmonella. Coarse-ground grains and access to litter have also been shown to decrease Salmonella shedding in poultry. The feed can also modify the gastro-intestinal tract microflora and influence the immune system, which can minimize Salmonella colonization and shedding. Feed additives, such as organic acids, short- and medium-chain fatty acids, probiotics, including competitive exclusion cultures, prebiotics and certain specific carbohydrates, such as mannan-based compounds, egg proteins, essential oils
Kitagawa, Ryo; Takaya, Akiko; Ohya, Mai; Mizunoe, Yoshimitsu; Takade, Akemi; Yoshida, Shin-ichi; Isogai, Emiko; Yamamoto, Tomoko
Gram-negative bacteria ubiquitously release membrane vesicles (MVs) into the extracellular milieu. Although MVs are the product of growing bacteria, not of cell lysis or death, the regulatory mechanisms underlying MV formation remained unknown. We have found that MV biogenesis is provoked by the induction of PagC, a Salmonella-specific protein whose expression is activated by conditions that mimic acidified macrophage phagosomes. PagC is a major constituent of Salmonella MVs, and increased ex...
Nugent, Staci L.; Meng, Fanhong; Martin, Gregory B; Altier, Craig
Salmonella remains a leading cause of bacterial food-borne disease, sickening millions each year. Although outbreaks of salmonellosis have traditionally been associated with contaminated meat products, recent years have seen numerous disease cases caused by the consumption of produce. Tomatoes have been specifically implicated, due to the ability of Salmonella spp. to enter the tomato fruit and proliferate within, making the decontamination of the raw product impossible. To investigate the ge...
Van Immerseel, Filip; Russell, James; Flythe, Michael; Gantois, Inne; Timbermont, Leen; Pasmans, Frank; Haesebrouck, Freddy; Ducatelle, Richard
Abstract Salmonella is a human pathogen that is commonly found in poultry products. It is possible to decrease chicken carcass and egg contaminations by adding organic acids to the feed or drinking water at appropriate times. Medium chain fatty acids are be more antibacterial against Salmonella than short-chain fatty acids. The antibacterial effect of these acids is species specific. Bacteria that are unable to decrease intracellular pH accumulate organic acid anions in accordance...
Jackson, Brendan R.; Griffin, Patricia M.; Cole, Dana; Walsh, Kelly A.; Chai, Shua J.
Salmonella enterica infections are transmitted not only by animal-derived foods but also by vegetables, fruits, and other plant products. To clarify links between Salmonella serotypes and specific foods, we examined the diversity and predominance of food commodities implicated in outbreaks of salmonellosis during 1998–2008. More than 80% of outbreaks caused by serotypes Enteritidis, Heidelberg, and Hadar were attributed to eggs or poultry, whereas >50% of outbreaks caused by serotypes Javiana...
Chowdry, N.; Threlfall, E J; Rowe, B.; Stanley, J.
An analysis of genotype was made for representative strains of Salmonella dublin. The collection consisted primarily of strains isolated from humans in England and Wales, and were of both intestinal and extra-intestinal origin. Three genetic elements were characterized by DNA hybridization. They were the spvBC genes, extrachromosomal virulence determinants, the salmonella-specific insertion sequence IS200, and the 16S ribosomal RNA genes, a phylogenetic marker. Two clones of S. dublin (SdRI a...
Löfström, Charlotta; Hansen, Flemming; Mansdal, Susanne;
Cost-effective and rapid monitoring of Salmonella in the meat production chain can contribute to food safety. The objective of this study was to validate an easy-to-use pre-PCR sample preparation method based on a simple boiling protocol for screening of Salmonella in meat and carcass swab samples...... deviating results were obtained (SP, SE, and AC were 100, 95, and 97%, respectively). This test is under implementation by the Danish meat industry, and can be useful for screening of large number of samples in the meat production, especially for fast release of minced meat with a short shelf life....
Foodborne pathogens are constantly adapting to circumvent intervention strategies. The ability to detect and overcome these adaptations are critical to ensure a safe food supply. We determined genotypic and/or phenotypic differences between Salmonella recovered from broiler chicks after comingling w...
Wegener, Henrik Caspar; Hald, Tine; Wong, Danilo Lo Fo;
We describe Salmonella control programs of broiler chickens, layer hens, and pigs in Denmark. Major reductions in the incidence of foodborne human salmonellosis have occurred by integrated control of farms and food processing plants. Disease control has been achieved by monitoring the herds and...
This podcast, featuring lead investigator Shauna Mettee, discusses the first known outbreak of Salmonella in people due to contact with water frogs. Created: 1/12/2010 by National Center for Zoonotic, Vector-Borne, and Enteric Diseases (NCZVED). Date Released: 1/12/2010.
Background: Since the early 1990’s there has been increasing awareness and concern regarding the development of antimicrobial resistance among bacteria of public health significance. Reports targeting zoonotic bacteria, and in particular Salmonella species, suggest that resistance is trending upwar...
Satish R Patil
Full Text Available Bacteria of the genus Salmonella are highly adapted for the growth in both humans and animals and cause a wide spectrum of disease. The growth of Serotypes S. typhi and S. paratyphi is restricted to human hosts, in whom these organisms cause enteric (typhoid fever. The remaining Serotypes (non typhoidal Salmonella or NTS can colonize the gastrointestinal tracts of the broad range of animals, including mammals, reptiles, birds and insects. The usual clinical presentation of non-typhoidal salmonellae (NTS infection is self limited gastroenteritis; however bacteremia and focal extra intestinal infection may occur. However salmonella localization to the skin presenting as cutaneous ulceration is regarded as a rare event. Rates of morbidity and mortality associated with NTS are highest among the elderly, infants, and immunocompromised individuals, including those with hemoglobinopathies, HIV infection, or infections that cause blockade of the reticuloendothelial system. We isolated S.infantis in 50 years old man with left leg cellulitis. The serotype was confirmed at Central Research Institute, Kasauli.
Wu, R; Felton, J
The Ames/Salmonella assay, developed by Professor Bruce Ames at the University of California, Berkeley, is a rapid and sensitive assay for detecting mutagenicity of various chemical compounds (Maron and Ames, 1983). It is a widely accepted short-term assay for detecting chemicals that induce mutations in the histidine (his) gene of Salmonella typhimurium. This is a reverse mutation assay that detects the mutational reversion of his-dependent Salmonella to the his-independent counterpart. Thereby, mutagenic compounds will increase the frequency of occurrence of his-independent bacterial colonies. The assay utilizes the specific genetically constructed strains of bacteria either with or without mammalian metabolic activation enzymes (S9), Aroclor induced rat liver homogenate to assess the mutagenicity of different compounds. In this study, we will use the Ames/Salmonella assay to investigate the mutagenicity of Hexanitrostilbene (HNS) from both Bofors and Pantex, and Dipicryethane (DPE).
The identification of Salmonella spp. from poultry meat was studied by comparing bacterial detection using the Gene-Trak colorimetric hybridization method, a PCR amplification kit and an Enzyme Linked Immunosorbent Assay (ELISA), and these methods were compared with the conventional methodology proposed by the United States Food and Drug Administration (US FDA) for detection of Salmonella in food samples. Forty positive and negative samples were studied. The three methods yielded similar results with levels of Salmonella greater than 10 CFU per sample, even when the samples were highly contaminated with competing bacteria. In contrast, 20 CFU of seed inoculum per sample was the lowest level of Salmonella detectable with all three methods and the standard culture method. The detection limits of the PCR and ELISA assays were 5 CFU/g after enrichment at 37 deg. C for 6 and 9 hours, respectively. Compared with conventional bacteriology, all three methods here demonstrated high sensitivity and specificity for Salmonella. (author)
Gradel, K.O.; Feld, Niels Christian; Andersen, J. S.
In the Danish Salmonella Control Program, eggs from broiler parent flocks are surveyed by serologic analysis every 4 wk for antibodies against Salmonella lipopolysaccharide O-antigens 1, 4, 5, 9, and 12 (Mix-enzyme-linked immunosorbent assay [ELISA]) and 6 and 7 (Infantis-ELISA). The antibody...... response is measured in percentage optical density (OD%) of a strong positive reaction, and the cutoff value has been determined to be 40 OD%. Two or more reactors above 40 OD% will place the parent flock under suspicion. There has been concern about possible cross-reactions between Salmonella spp....... and other Enterobacteriaceae, e.g., Escherichia coli, because a high specificity of a Salmonella antibody test is desirable. Moreover, false-positive Salmonella results have economic consequences and impede planning the production. A case-control study based on cases of clinical E. coli infections...
Full Text Available Abstract Salmonellosis is the main cause of human bacterial gastroenteritis in most European countries. Infections with Salmonella is usually subclinical, whereas clinical cases show symptoms with a wide range of severity. Infection is most commonly associated with the consumption of meat, especially poultry or pork, and eggs and their products. Salmonella can enter the food chain at any point throughout its length. The principal reservoir of Salmonellae is the gastrointestinal tract of mammals and birds, but Salmonellae are able to survive and even multiply in many external environments. In Norway, Sweden and Finland cost effective prevention methods have been used for several years to prevent and control Salmonellea infections. In addition, competitive exclusion (CE and vaccination might be relevant as biological methods to prevent colonisation of bird intestines by enteropathogens, especially Salmonella. Antibiotic drug resistance has been a problem since the start of the antibiotic era. The cause for anxiety is that more and more bacteria are becoming resistant, often to a whole range of antibiotics. The debate on the use of antimicrobials in veterinary medicine and animal production dates back almost as long as the use itself. There is a clear evidence to show that antibacterial agents given to animals for growth promotion, prophylactic purposes or treatment induce a rise in the number of antibiotic resistant strains isolated from the animals. These bacteria may be transmitted to humans by several possible routes. There are thus strong arguments for preventive efforts which have to be directed towards identifying real critical control points (HACCP throughout the whole food chain, which starts from the farm and ends at the consumer's table.
McDougal, D L; Treleaven, B E; Renshaw, E C
A new Salmonella serotype, Salmonella enteritidis serotype Grandhaven (30(1):r:1,2), was isolated from the stool of a 35-year-old man with mild gastroenteritis. He had just returned from Sudan, Africa.
Olsson, M; Syk, A; Wollin, R
We have tested 750 Salmonella strains and 130 strains of other species of the family Enterbacteriaceae with the 4-methylumbelliferyl caprilate reagent (MUCAP) test. The MUCAP test is a fluorescence test for rapid identification of Salmonella strains. The non-Salmonella strains were strains sent for identification as suspected Salmonella strains and thus have phenotypes similar to those of Salmonella strains. All 748 tested Salmonella strains of subgroups I, II, III, and IV were positive in th...
Switt, Andrea I. Moreno; den Bakker, Henk C.; Vongkamjan, Kitiya; Hoelzer, Karin; Warnick, Lorin D.; Cummings, Kevin J.; Wiedmann, Martin
Salmonella is an animal and human pathogen of worldwide concern. Surveillance programs indicate that the incidence of Salmonella serovars fluctuates over time. While bacteriophages are likely to play a role in driving microbial diversity, our understanding of the ecology and diversity of Salmonella phages is limited. Here we report the isolation of Salmonella phages from manure samples from 13 dairy farms with a history of Salmonella presence. Salmonella phages were isolated from 10 of the 13...
Addwebi, Tarek M; Call, Douglas R; Shah, Devendra H
Salmonella enterica serovar Enteritidis is one of the most common serovars associated with poultry and poultry product contamination in the United States. We previously identified 14 mutant strains of Salmonella Enteritidis phage type 4 (PT4) with significantly reduced invasiveness in human intestinal epithelial cells (Caco-2), chicken macrophages (HD-11), and chicken hepatocellular epithelial cells (LMH). These included Salmonella Enteritidis mutants with transposon insertions in 6 newly identified Salmonella Enteritidis-specific genes (pegD and SEN1393), and genes or genomic islands common to most other Salmonella serovars (SEN0803, SEN0034, SEN2278, and SEN3503) along with 8 genes previously known to contribute to enteric infection (hilA, pipA, fliH, fljB, csgB, spvR, and rfbMN). We hypothesized that Salmonella Enteritidis employs both common Salmonella enterica colonization factors and Salmonella Enteritidis-specific traits to establish infection in chickens. Four Salmonella Enteritidis mutants (SEN0034::Tn5, fliH::Tn5, SEN1393::Tn5, and spvR::Tn5) were indistinguishable from the isogenic wild-type strain when orally inoculated in 1-d-old chickens, whereas 2 mutants (CsgB::Tn5 and PegD::Tn5) were defective for intestinal colonization (P pipA::Tn5) showed significant in vivo attenuation in more than one organ (P < 0.05). Complementation studies confirmed the role of rfbN and SEN3503 during infection. This study should contribute to a better understanding of the mechanisms involved in Salmonella Enteritidis pathogenesis, and the target genes identified here could potentially serve as targets for the development of live-attenuated or subunit vaccine. PMID:24706964
Oral priming with Salmonella Typhi vaccine strain CVD 909 followed by parenteral boost with the S. Typhi Vi capsular polysaccharide vaccine induces CD27+IgD-S. Typhi-specific IgA and IgG B memory cells in humans.
Wahid, Rezwanul; Pasetti, Marcela F; Maciel, Milton; Simon, Jakub K; Tacket, Carol O; Levine, Myron M; Sztein, Marcelo B
Attenuated live oral typhoid vaccine candidate CVD 909 constitutively expresses Salmonella Typhi capsular polysaccharide antigen (Vi). A randomized, double-blind, heterologous prime-boost clinical study was conducted to determine whether immunity to licensed parenteral Vi vaccine could be enhanced by priming with CVD 909. Priming with CVD 909 elicited higher and persistent, albeit not significant, anti-Vi IgG and IgA following immunization with Vi, than placebo-primed recipients. Vi-specific IgA B memory (B(M)) cells were significantly increased in CVD 909-primed subjects. S. Typhi-specific LPS and flagella IgA B(M) cells were observed in subjects immunized with CVD 909 or with the licensed Vi-negative oral typhoid vaccine Ty21a. CVD 909-induced B(M) cells exhibited a classical B(M) phenotype (i.e., CD3(-)CD19(+)IgD(-)CD27(+)). This is the first demonstration of classical B(M) cells specific for bacterial polysaccharide or protein antigens following typhoid immunization. The persistent IgA B(M) responses demonstrate the capacity of oral typhoid vaccines to prime mucosally relevant immune memory. PMID:21146460
In many countries the incidence of human salmonella infections has markedly increased in recent years. To discuss recent developments and current understanding on the control of salmonella infections in animals, WHO organized a Consultation on the Control of Salmonella Infections in Animals: Prevention of Foodborne Salmonella Infections in Humans, held in Jena, Germany, on 21-26 November 1993. The present article summarizes the recommendations made by the participants on the pathoimmunogenesi...
Invasão de áreas de savana intra-amazônicas por Digitonthophagus gazella (Fabricius, 1787 (Insecta: Coleoptera: Scarabaeidae Invasion of intra-Amazonian savannas by the dung beetle Digintonthophagus gazella (Fabricius, 1787 (Insecta: Coleoptera: Scarabaeidae
Rodrigo Augusto Matavelli
Full Text Available Neste estudo, foi avaliada a invasão da comunidade de Scarabaeinae detritívoros de uma savana amazônica pela espécie africana Digitonthophagus gazella (Fabricius 1787. O estudo foi realizado nas proximidades da vila de Alter do Chão (2º 31' S e 55º 00' W, localizada a aproximadamente 36 km a sudoeste de Santarém, Pará, Brasil. Vinte e duas áreas de savanas de 3,75 ha (250 x 150 m distribuídas em 30.000 ha foram amostradas, no período de 21 de julho a 13 de agosto de 2003, utilizando 66 armadilhas de queda com três tipos de iscas (fezes bovinas, fezes humanas e carcaças. Foram encontrados indivíduos de D. gazella em quatro das vinte e duas áreas amostradas. Procurou-se explicar a presença da espécie nas áreas de savana através de análises de regressão logística, onde as variáveis explicativas foram: ocorrência de queimada nos últimos seis anos, diversidade e abundância total de Scarabaeidae nativos presentes na área, abundância de Canthon sp.1, (espécie de Scarabaeidae mais abundante na região. Exceto pela abundância total de indivíduos de Scarabaeidae nativos, nenhuma das variáveis bióticas e abióticas tiveram efeito estatisticamente significativo na presença do D. gazella. Estes resultados podem ser explicados por: (a algum fator ainda não analisado, relacionado à invasão da área pelo D. gazella; (b Não houve tempo para a dispersão e estabelecimento da espécie em todas as áreas; (c A comunidade nativa de Scarabaeinae apresenta resistência à invasão pelo D. gazella.This work aimed to verify the invasion of the dung beetles community from intra-Amazonian savanna by the African species Digitonthophagus gazella (Fabricius 1787. The research was carried out near Alter do Chão village (2º 31' S; 55º 00' W, 36 Km Southwest of Santarém, Pará, Brazil. Twenty two areas were sampled, from 21 July to 13 August 2003, using 66 baited pitfall traps (cattle dung, human faeces and carcass bait. D. gazella
Shima Shaigan nia
Conclusions: To our best knowledge the present study is the first prevalence report of Salmonella spp., Salmonella enteritidis and Salmonella typhimurium in raw sheep and goat samples in Iran. Consumption of pasteurized milk and dairy products can reduce the risk of salmonellosis.
Influência da invasão tumoral da linha de anastomose na sobrevivência de pacientes com câncer de coto gástrico The influence of tumor invasion in anastomotic line on survival of patient with gastric stump cancer
Ana Lúcia Granja Scarabel Nogueira Carrasco; Cláudio José Caldas Bresciani; Rodrigo Oliva Perez; Bruno Zilberstein; Ivan Cecconello
RACIONAL: O câncer do coto gástrico desenvolve- se no remanescente gástrico de gastrectomia realizada há pelo menos 5 anos por doença benigna e os sítios mais comuns de acometimento são próximo à anastomose e na pequena curvatura. Considera-se que o coto gástrico é estado pré-canceroso. OBJETIVOS: Identificar o padrão de disseminação de linfonodos acometidos, quantificar a invasão tumoral da linha de anastomose e correlacionar: a invasão da linha de anastomose com o comprometimento linfonodal...
Kobra Salimian Rizi
Full Text Available Background and Objectives: Salmonella is an important food-borne pathogen in humans. Strains of Salmonella spp. That producing extended-spectrum β-Lactamases have become a concern in medicine regarding both antimicrobial treatment and infection control program. The objective of this study was to describe the antibiotic susceptibility, ESBL production and determining the prevalence of the blaCTX-M-1 group among clinical isolates of Salmonella spp.Materials and Methods: A total of 110 Salmonella isolates collected from four Tehran hospitals during May 2012 and April 2013. The specific monovalan Salmonella antisera were used for serogrouping of Salmonella isolates. Antibacterial susceptibility was determined by disk diffusion and ESBL phenotype was confirmed by combination disk method. The blaCTX-M-1 group was identified by PCR with specific primers. The transferability of the blaCTX-1 group was tested by conjugation with broth matting method.Results: The prevalence of Salmonella serogroups consist of 56.4% serogroup D, 13.6 % serogroup C, 10 % serogroup B, and 1.8 % serogroup A and 18.2% other serogroups. Maximal resistance in Salmonella isolates was noticed against trimethoprim-sulfamethoxazole (63.6% and nalidixic-acid (47/3%. All isolates were susceptible to imipenem and ciprofloxacin. Four isolates (3.6% showed ESBLs phenotype. All Salmonella spp. that produce ESBls have blaCTX-1 genes group. A conjugative plasmid containing blaCTX-1 group was found in one Salmonella isolate.Conclusion: This study demonstrates the predominant presence of the gene encoding CTX-M-1 group among ESBLs producing of Salmonella spp. They can transmit to bacteria of this genus or even other genera of enteric bacteria.Keywords: Salmonella spp., blaCTX-M-1 group, antibiotic resistance, conjugation, broth matting
Pochop, Jaroslav; Kačániová, Miroslava; Hleba, Lukáš; Lejková, Jadža; Fikselová, Martina; Kunová, Simona; Kluz, Maciej
The aim of this study was to follow contamination of ready to eat milk and meat products with Salmonella spp. by using the StepOne real-time polymerase chain reaction (PCR). Classical microbiological methods for detection of foodborne bacteria involve the use of pre-enrichment and/or specific enrichment, following isolation of bacteria in solid media and the final confirmation by biochemical and/or serological tests. We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and MicroSEQ® Salmonella spp. Detection Kit for pursuance of the real-time PCR (Applied Biosystems). In samples without incubation we detected strain of Salmonella sp. in 5 out of 25 samples (swabs), as well as in the internal positive control (IPC), which was positive in all samples. This StepOne real-time PCR assay is extremely useful for any laboratory equipped by real-time PCR. It is a fast, reproducible, simple, specific and sensitive way to detect nucleic acids, which could be used in clinical diagnostic tests in the future. Our results indicated that real-time PCR assay developed in this study could sensitively detect Salmonella spp. in ready-to-eat food. This could prevent infection caused by Salmonella, and also could benefit food manufacturing companies by extending their product's shelf-life as well as saving the cost of warehousing their food products while awaiting pathogen testing results. PMID:21879831
Cao, Guojie; Allard, Marc; Strain, Errol; Stones, Robert; Zhao, Shaohua; Brown, Eric; Meng, Jianghong
Salmonella enterica subspecies enterica serotype Newport is one of the common serotypes causing foodborne salmonellosis outbreaks in the United States. Salmonella Newport consists of three lineages exhibiting extensive genetic diversity. Due to the importance of Salmonella pathogenicity islands 5 and 6 (SPI-5 and SPI-6) in virulence of pathogenic Salmonella, the genetic diversity of these two SPIs may relate to different potentials of Salmonella Newport pathogenicity. Most Salmonella Newport strains from North America belong to Salmonella Newport lineages II and III. A total 28 Salmonella Newport strains of lineages II and III from diverse sources and geographic locations were analyzed, and 11 additional Salmonella genomes were used as outgroup in phylogenetic analyses. SPI-5 was identified in all Salmonella Newport strains and 146 single nucleotide polymorphisms (SNPs) were detected. Thirty-nine lineage-defining SNPs were identified, including 18 nonsynonymous SNPs. Two 40-kb genomic islands (SPI5-GI1 and SPI5-GI2) encoding bacteriophage genes were found between tRNA-ser and pipA. SPI5-GI1 was only present in Salmonella Newport multidrug-resistant strains of lineage II. SPI-6 was found in all strains but three Asian strains in Salmonella Newport lineage II, whereas the three Asian strains carried genomic island SPI6-GI1 at the same locus as SPI-6 in other Salmonella. SPI-6 exhibited 937 SNPs, and phylogenetic analysis demonstrated that clustering of Salmonella Newport isolates was a reflection of their geographic origins. The sequence diversity within SPI-5 and SPI-6 suggests possible recombination events and different virulence potentials of Salmonella Newport. The SNPs could be used as biomarkers during epidemiological investigations. PMID:25188010
Iwu, Chinwe Juliana; Iweriebor, Benson Chuks; Obi, Larry Chikwelu; Basson, Albertus Kotze; Okoh, Anthony Ifeanyi
The exposure of farm animals to antimicrobials for treatment, prophylaxis, or growth promotion can select for resistant bacteria that can be transmitted to humans, and Salmonella as an important zoonotic pathogen can act as a potential reservoir of antimicrobial resistance determinants. We assessed the antibiogram profiles of Salmonella species isolated from pig herds in two commercial farms in South Africa. Two hundred fifty-eight presumptive Salmonella isolates were recovered from the fecal samples of 500 adult pigs. Specific primers targeting Salmonella serogroups A, B, C1, C2, and D were used to determine the prevalence of different serogroups. Only serogroup A (n = 48) was detected, while others were not. Antimicrobial susceptibility of the confirmed Salmonella serogroup A isolates was performed by using the disk diffusion method against a panel of 18 antibiotics. All the 48 isolates were resistant to tetracycline and oxytetracycline, while 75% were resistant to ampicillin, sulphamethoxazole-trimethoprim, nalidixic acid, and streptomycin. All the isolates exhibited multidrug resistance, with the predominant phenotype being against 11 antibiotics, and multiple antibiotic resistance index ranged between 0.3 and 0.6. The incidence of genes encoding resistance against ampicillin (ampC), tetracycline (tetA), and streptomycin (strA) were 54, 61, and 44%, respectively. We conclude that healthy pigs are potential reservoirs of multidrug-resistant Salmonella that could be transmitted to humans through the food chain and, hence, a significant public health threat. PMID:27357044
The use of wastewater for irrigation may introduce antimicrobials and human pathogens into the food supply through vegetative uptake. The objective of this study was to investigate the uptake of three antimicrobials and Salmonella in two lettuce cultivars. After repeated subirrigation with synthetic wastewater, lettuce leaves and soil were collected at three sequential harvests. The internalization frequency of Salmonella in lettuce was low. A soil horizon-influenced Salmonella concentration gradient was determined with concentrations in bottom soil 2 log CFU/g higher than in top soil. Lincomycin and sulfamethoxazole were recovered from lettuce leaves at concentrations as high as 822 ng/g and 125 ng/g fresh weight, respectively. Antimicrobial concentrations in lettuce decreased from the first to the third harvest suggesting that the plant growth rate may exceed antimicrobial uptake rates. Accumulation of antimicrobials was significantly different between cultivars demonstrating a subspecies level variation in uptake of antibiotics in lettuce. - Highlights: • Antimicrobial uptake in lettuce is cultivar dependent. • Antimicrobial concentrations in lettuce decrease despite repeated exposure. • Lincomycin is better conserved in the soil-plant system than oxytetracycline or sulfamethoxazole. • Subirrigation resulted in more Salmonella in bottom soil than in top soil. • Internalization frequency of Salmonella in lettuce is low despite repeated exposure. - Cultivar-specific differences in lincomycin and sulfamethazine uptake were observed in lettuce, while uptake of Salmonella was low despite repeated exposure from wastewater
Durul, Bora; Acar, Sinem; Bulut, Ece; Kyere, Emmanuel O; Soyer, Yeşim
Salmonella is commonly found in a variety of food products and is a major cause of bacterial foodborne illness throughout the world. In this study, we investigated the prevalence and diversity of Salmonella in eight different food types: sheep ground meat, cow ground meat, chicken meat, cow offal, traditional Sanliurfa cheese, unripened feta cheese, pistachios, and isot (a spice blend of dried red peppers specific to Sanliurfa), traditionally and commonly consumed in Turkey. Among 192 food samples, Salmonella was detected in 59 samples, with the highest prevalence in raw poultry parts (58%) and offal (58%) samples, while Salmonella was not detected in pistachios and dried red pepper. Resultant Salmonella isolates were characterized by serotyping, multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). Ten different serotypes represented 10 MLST sequence types (STs) with 1 novel ST and 17 PFGE types. Antimicrobial resistance profiling revealed that 30.5% of the isolates were resistant to two or more antimicrobials. Salmonella enterica subsp. enterica serotype Telaviv, which is rare throughout the world, was the second most common serotype isolated from food samples in this study, suggesting that this serotype might be one of the subtypes that is endemic to Turkey. PMID:26489049
Full Text Available Salmonella is a globally widespread food-borne pathogen having major impact on public health. All motile serovars of Salmonella enterica of poultry origin are zoonotic, and contaminated meat and raw eggs are an important source to human infections. Information on the prevalence of Salmonella at farm/holding level, and the zoonotic serovars circulating in layer poultry in the South and South-East Asian countries including Bangladesh, where small-scale commercial farms are predominant, is limited. To investigate the prevalence of Salmonella at layer farm level, and to identify the prevalent serovars we conducted a cross-sectional survey by randomly selecting 500 commercial layer poultry farms in Bangladesh. Faecal samples from the selected farms were collected following standard procedure, and examined for the presence of Salmonella using conventional bacteriological procedures. Thirty isolates were randomly selected, from the ninety obtained from the survey, for serotyping and characterized further by plasmid profiling and pulsed-field gel electrophoresis (PFGE. Results of the survey showed that the prevalence of motile Salmonella at layer farm level was 18% (95% confidence interval 15-21%, and Salmonella Kentucky was identified to be the only serovar circulating in the study population. Plasmid analysis of the S. Kentucky and non-serotyped isolates revealed two distinct profiles with a variation of two different sizes (2.7 and 4.8 kb. PFGE of the 30 S. Kentucky and 30 non-serotyped isolates showed that all of them were clonally related because only one genotype and three subtypes were determined based on the variation in two or three bands. This is also the first report on the presence of any specific serovar of Salmonella enterica in poultry in Bangladesh.
Roof, D M; Roth, J R
Ethanolamine can serve as the sole source of carbon and nitrogen for Salmonella typhimurium if vitamin B12 is present to serve as a cofactor. The pathway for ethanolamine utilization has been investigated in order to understand its regulation and determine whether the pathway is important to the selective forces that have maintained the ability to synthesize B12 in S. typhimurium. We isolated mutants that are defective in ethanolamine utilization (eut mutants). These mutants defined a cluster...
Čanović Predrag S.
Full Text Available Two patients with Reiter's syndrome, after Salmonella infection were treated on the Infections disease ward at Clinical hospital center in Kragujevac. In the first patient, ten days after the onset of Salmonella infection, signs of edema and pain in the right ankle occurred, accompanied by expressed conjunctivitis. Within next two months consecutive metatarsophalanges changes joint of the right foot have appeared. In the second patient, two weeks after the onset of Salmonella infection, edema of the left hand joints and a week later edema of the right hand and right ankle joints appeared. In both patients inflammatory syndrome was expressed (high erythrocyte sedimentation rates, fibrinogen, C-reactive protein along with negative rheumatoid factors and positive antigen HLA-B27. Outcome of the disease in both cases was favorable upon receiving nonsteroid antirheumatic therapy. Signs of arthritis disappeared after three months. No signs of recurrent arthritis have been seen during the next four years in the first and next two years in the second patient.
Charles L. Hofacre
Full Text Available Accurate and fast detection of harmful Salmonella is a major concern of food safety. Common Salmonella serotypes responsible for human associated foodborne outbreaks are S. Enteritidis, S. Hadar, S. Heidelberg, and S. Typhimurium are also commonly isolated from poultry. Serology is commonly used to monitor disease in poultry, therefore application of Salmonella serotype-specific test will have added value in Salmonella surveillance or monitoring vaccine efficacy. Recombinant flagellins were purified to be used as antigens in an ELISA. In this study, an ELISA was developed for the serological detection of S. Enteritidis. Once optimized, 500 ng of purified recombinant S. Enteritidis flagellin and a 1:64 dilution were determined to be optimal for testing sera. A negative baseline cutoff was calculated to be an optical density (OD of 0.35. All sera from birds with history of S. Enteritidis exposure tested positive and all sera from chickens with no exposure tested negative to this Salmonella serotype. Current ELISA for serological detection of Salmonella suffers from cross reactivity inherent in lipopolysaccharide (LPS or whole cell antigen based serological tests. This new ELISA eliminates common cross reactivity by focusing specifically on the flagellins of the Salmonella serotypes common in poultry and associated with foodborne outbreaks.
Korver H; Raes M; Maas HME; Ward LR; Wannet WJB; Henken AM; MGB; LIS
Test resultaten van Salmonella sero- en faagtypering en antimicrobiele gevoeligheidsbepalingen door de Nationale Referentie Laboratoria voor Salmonella in de Lidstaten van de Europese Unie en EnterNet Laboratoria: Ringonderzoek VI (2001) voor Salmonella. Een zesde ringonderzoek betreffende de typering van Salmonella werd georganiseerd door het Communautair Referentie Laboratorium voor Salmonella (CRL-Salmonella, Bilthoven, Nederland) in samenwerking met Public Health Laboratory Services (PHLS...
Marvy Khrisna Pranamartha
Full Text Available ABSTRAK Demam tifoid disebabkan oleh bakteri Salmonella typhi, dengan gejala umum berupa demam tinggi dan nyeri perut. Tifoid adalah penyakit infeksi yang disebabkan oleh bakteri Salmonella typhi, yang masuk ke dalam tubuh melalui mulut dan saluran cerna.1 Untuk bisa memahami patogenesis dari demam tifoid sampai ke tingkat selular dan molekular, ada 5 hal penting yang harus digaris bawahi, yaitu: 1.\tTipe 3 Sistem Sekresi (T3SS 2.\tVirulence Genes dari Salmonella yang mengkode 5 SIP (Salmonella Invasion Protein SIP A, B, C, D, dan E. 3.\tToll R2 dan toll R3 yang merupakan lapisan luar dari makrofag. 4.\tSistem imun lumen usus sampai ke organ dalam 5.\tFungsi endotelial sel dalam inflamasi. Infeksi Salmonella dapat berakibat fatal kepada bayi, balita, ibu hamil dan kandungannya serta orang lanjut usia. Hal ini disebabkan karena kekebalan tubuh mereka yang menurun. Virulensi salmonella tidak lepas dari peranan SPI, yang terletak di dalam kromosom dan plasmid bakteri. Dimana SPI 1 dan SPI 2 telah dikaji cukup mendalam karena keterkaitannya dengan T3SS, dan berperan sangat penting pada invasi awal serta siklus hidup intrasel dari bakteri Salmonella. Kontaminasi Salmonella dapat dicegah dengan mencuci tangan dan menjaga kebersihan makanan yang dikonsumsi. Selalu menjaga kebersihan lingkungan hidup kita agar terhindar dari kontaminasi dengan bakteri Salmonella typhi. Agar mewaspadai sejak dini pencegahan dan pengobatan penyakit typhus. Studi mendalam perlu dilakukan agar kita mampu lebih memahami proses kompleks antara patogen dan sel inang. Mengingat dari 15 SPI yang sudah diketahui, hanya SPI 1 dan SPI 2 yang sudah dikaji secara mendalam. Kata Kunci: Salmonella, Salmonella Invasion Protein, Typhi.
ROBERTO GOMES NOGUEIRA
Full Text Available Descrevemos o caso de um paciente com diagnóstico de germinoma nos núcleos da base e tálamo, com invasão do tronco cerebral, ressaltando as características observadas nos exames de tomografia computadorizada e de ressonância magnética, os possíveis diagnósticos diferenciais, e a necessidade da comprovação anátomo-patológica, por tratar-se de localização pouco frequente de um tumor com possibilidade de evolução favorável após tratamento com quimio e radioterapia.We report an unusual case of germinoma arising from the basal ganglia and thalamus with brain stem invasion, with emphasis on computed tomography and magnetic resonance findings. Diagnosis was confirmed by histopathologic examination. Early detection of this tumor is important due to its potential response to treatment.
Singhal, Vikram; EK, Saleem; SM, Rajesh; Coutinho, Anita
Salmonella meningitis is rarely seen in neonates and if it is present, it is associated with complications like subdural collections and abscesses. We are presenting a case of Salmonella meningitis in a 28 days old neonate who developed complications like extensive subdural collections and ventriculitis. The child responded well to prolonged systemic antibiotics and other supportive measures.
Salmonella Heidelberg is the fourth-most commonly reported Salmonella serotype to cause human illness. There have been several outbreaks and recalls caused by S. Heidelberg in ready to eat foods. Recently, 700 people became ill from ingesting hummus shirazi contaminated with S. Heidelberg. This stud...
Yan, Jing-Jou; Chiou, Chien-Shun; Lauderdale, Tsai-Ling Yang; Tsai, Shu-Huei; Wu, Jiunn-Jong
We report the prevalence and characteristics of Salmonella strains resistant to ciprofloxacin and extended-spectrum cephalosporins in Taiwan from January to May 2004. All isolates resistant to extended-spectrum cephalosporins carried bla CMY-2, and all ciprofloxacin-resistant Salmonella enterica serotype Choleraesuis isolates were genetically related.
... biological product containing Salmonella dublin fraction shall meet the applicable requirements in 9 CFR 113... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Salmonella Dublin Bacterin. 113.123... Inactivated Bacterial Products § 113.123 Salmonella Dublin Bacterin. Salmonella Dublin Bacterin shall...
... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Salmonella Typhimurium Bacterin. 113... REQUIREMENTS Inactivated Bacterial Products § 113.120 Salmonella Typhimurium Bacterin. Salmonella Typhimurium Bacterin shall be prepared from a culture of Salmonella typhimurium which has been inactivated and...
Non-typhoidal salmonella bacteremia may result in extra gastrointestinallocalization of infection. Aortitis due to non-typhoidal salmonella wasreported to be the cause of 38-42% of all infected abdominal aortitis.Underlying atherosclerosis is a frequent site for salmonella aortitis. Wedescribe here a case of possible salmonella aortitis in a renal transplantpatient. (author)
Jensen, Annette Nygaard
Forsøg har vist at Salmonella bakterier kan overføres fra smittede økogrise til salmonella-frie grise holdt på udendørs arealer. Desuden kan salmonella bakterier overleve i foldmiljøet og under visse betingelser smitte salmonella-frie grise, som senere indsættes i folden.
Full Text Available Salmonella enterica species is a Gram negative bacterium, which is responsible for a wide range of food- and water-borne diseases in both humans and animals, thereby posing a major threat to public health. Recently, there has been an increasing number of reports, linking Salmonella contaminated raw vegetables and fruit with food poisoning. Many studies have shown that an essential feature of the pathogenicity of Salmonella is its capacity to cross a number of barriers requiring invasion of a large variety of cells and that the extent of internalization may be influenced by numerous factors. However, it is poorly understood how Salmonella successfully infects hosts as diversified as animals or plants. The aim of this review is to describe the different stages required for Salmonella interaction with its hosts: (i attachment to host surfaces; (ii entry processes; (iii, multiplication; (iv suppression of host defence mechanisms ; and to point out similarities and differences between animal and plant infections.
Prophylactic Administration of Vector-Encoded Porcine Granulocyte-Colony Stimulating Factor Reduces Salmonella Shedding, Tonsil Colonization, and Microbiota Alterations of the Gastrointestinal Tract in Salmonella-Challenged Swine
Bearson, Shawn M. D.; Bearson, Bradley L.; Loving, Crystal L.; Allen, Heather K.; Lee, InSoo; Madson, Darin; Kehrli, Marcus E.
Salmonella colonization of food animals is a concern for animal health and public health as a food safety risk. Various obstacles impede the effort to reduce asymptomatic Salmonella carriage in food animals, including the existence of numerous serovars and the ubiquitous nature of Salmonella. To develop an intervention strategy that is non-specific yet effective against diverse Salmonella serovars, we explored the prophylactic use of a cytokine to decrease Salmonella in swine by boosting the host’s innate immune system. Granulocyte-colony stimulating factor (G-CSF) is the major cytokine regulating the production, differentiation, function, and survival of neutrophils. Neutrophils play a critical role in the response to Salmonella; therefore, we evaluated the vectored-delivery of porcine G-CSF as a prophylactic to reduce Salmonella in pigs. Crossbred pigs, 5 weeks of age, were intramuscularly injected with a replication-defective human adenovirus (Ad5) engineered to express porcine G-CSF (Ad5-G-CSF, n = 9). Control pigs received the same Ad5 vector lacking the gene encoding G-CSF (Ad5-empty, n = 7). Four days later, all pigs (n = 16) were intranasally inoculated with 1 × 107 colony forming unit (CFU) of Salmonella enterica serovar Typhimurium UK1. At 2 and 3 days post-challenge with Salmonella, Ad5-G-CSF-treated pigs shed significantly less Salmonella (~103 CFU/g) in their feces than Ad5-empty-treated pigs (~104–105 CFU/g; P structure of Salmonella-challenged pigs was less disturbed post-challenge in the Ad5-G-CSF-treated pigs than the Ad5-empty-treated pigs. This suggests that Ad5-G-CSF administration mitigated changes in the microbial community structure caused by Salmonella challenge. Collectively, these data suggest that delivery of a targeted immunostimulant to enhance neutropoiesis may be a strategy to reduce Salmonella colonization, potentially during periods of immunological stress. PMID:27610361
Dagmara I Kisiela
Full Text Available Whereas the majority of pathogenic Salmonella serovars are capable of infecting many different animal species, typically producing a self-limited gastroenteritis, serovars with narrow host-specificity exhibit increased virulence and their infections frequently result in fatal systemic diseases. In our study, a genetic and functional analysis of the mannose-specific type 1 fimbrial adhesin FimH from a variety of serovars of Salmonella enterica revealed that specific mutant variants of FimH are common in host-adapted (systemically invasive serovars. We have found that while the low-binding shear-dependent phenotype of the adhesin is preserved in broad host-range (usually systemically non-invasive Salmonella, the majority of host-adapted serovars express FimH variants with one of two alternative phenotypes: a significantly increased binding to mannose (as in S. Typhi, S. Paratyphi C, S. Dublin and some isolates of S. Choleraesuis, or complete loss of the mannose-binding activity (as in S. Paratyphi B, S. Choleraesuis and S. Gallinarum. The functional diversification of FimH in host-adapted Salmonella results from recently acquired structural mutations. Many of the mutations are of a convergent nature indicative of strong positive selection. The high-binding phenotype of FimH that leads to increased bacterial adhesiveness to and invasiveness of epithelial cells and macrophages usually precedes acquisition of the non-binding phenotype. Collectively these observations suggest that activation or inactivation of mannose-specific adhesive properties in different systemically invasive serovars of Salmonella reflects their dynamic trajectories of adaptation to a life style in specific hosts. In conclusion, our study demonstrates that point mutations are the target of positive selection and, in addition to horizontal gene transfer and genome degradation events, can contribute to the differential pathoadaptive evolution of Salmonella.
Feldsine, Philip; Kaur, Mandeep; Shah, Khyati; Immerman, Amy; Jucker, Markus; Lienau, Andrew
Assurance GDSTM for Salmonella Tq has been validated according to the AOAC INTERNATIONAL Methods Committee Guidelines for Validation of Microbiological Methods for Food and Environmental Surfaces for the detection of selected foods and environmental surfaces (Official Method of AnalysisSM 2009.03, Performance Tested MethodSM No. 050602). The method also completed AFNOR validation (following the ISO 16140 standard) compared to the reference method EN ISO 6579. For AFNOR, GDS was given a scope covering all human food, animal feed stuff, and environmental surfaces (Certificate No. TRA02/12-01/09). Results showed that Assurance GDS for Salmonella (GDS) has high sensitivity and is equivalent to the reference culture methods for the detection of motile and non-motile Salmonella. As part of the aforementioned validations, inclusivity and exclusivity studies, stability, and ruggedness studies were also conducted. Assurance GDS has 100% inclusivity and exclusivity among the 100 Salmonella serovars and 35 non-Salmonella organisms analyzed. To add to the scope of the Assurance GDS for Salmonella method, a matrix extension study was conducted, following the AOAC guidelines, to validate the application of the method for selected spices, specifically curry powder, cumin powder, and chili powder, for the detection of Salmonella. PMID:26268975
Sharon M Tennant
Full Text Available Certain serovars of Salmonella enterica subsp. enterica cause invasive disease (e.g., enteric fever, bacteremia, septicemia, meningitis, etc. in humans and constitute a global public health problem. A rapid, sensitive diagnostic test is needed to allow prompt initiation of therapy in individual patients and for measuring disease burden at the population level. An innovative and promising new rapid diagnostic technique is microwave-accelerated metal-enhanced fluorescence (MAMEF. We have adapted this assay platform to detect the chromosomal oriC locus common to all Salmonella enterica subsp. enterica serovars. We have shown efficient lysis of biologically relevant concentrations of Salmonella spp. suspended in bacteriological media using microwave-induced lysis. Following lysis and DNA release, as little as 1 CFU of Salmonella in 1 ml of medium can be detected in <30 seconds. Furthermore the assay is sensitive and specific: it can detect oriC from Salmonella serovars Typhi, Paratyphi A, Paratyphi B, Paratyphi C, Typhimurium, Enteritidis and Choleraesuis but does not detect Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae or Acinetobacter baumanii. We have also performed preliminary experiments using a synthetic Salmonella oriC oligonucleotide suspended in whole human blood and observed rapid detection when the sample was diluted 1:1 with PBS. These pre-clinical data encourage progress to the next step to detect Salmonella in blood (and other ordinarily sterile, clinically relevant body fluids.