JoVE Visualize What is visualize?
Stop Reading. Start Watching.
Advanced Search
Stop Reading. Start Watching.
Regular Search
Find video protocols related to scientific articles indexed in Pubmed.
The role of ClpP, RpoS and CsrA in growth and filament formation of Salmonella enterica serovar Typhimurium at low temperature.
BMC Microbiol.
PUBLISHED: 08-14-2014
Show Abstract
Hide Abstract
Salmonellae are food-borne pathogens of great health and economic importance. To pose a threat to humans, Salmonellae normally have to cope with a series of stressful conditions in the food chain, including low temperature. In the current study, we evaluated the importance of the Clp proteolytic complex and the carbon starvation protein, CsrA, for the ability of Salmonella Typhimurium to grow at low temperature.
Related JoVE Video
Identification of potential drug targets in Salmonella enterica sv. Typhimurium using metabolic modelling and experimental validation.
Microbiology (Reading, Engl.)
PUBLISHED: 04-28-2014
Show Abstract
Hide Abstract
Salmonella enterica sv. Typhimurium is an established model organism for Gram-negative, intracellular pathogens. Owing to the rapid spread of resistance to antibiotics among this group of pathogens, new approaches to identify suitable target proteins are required. Based on the genome sequence of S. Typhimurium and associated databases, a genome-scale metabolic model was constructed. Output was based on an experimental determination of the biomass of Salmonella when growing in glucose minimal medium. Linear programming was used to simulate variations in the energy demand while growing in glucose minimal medium. By grouping reactions with similar flux responses, a subnetwork of 34 reactions responding to this variation was identified (the catabolic core). This network was used to identify sets of one and two reactions that when removed from the genome-scale model interfered with energy and biomass generation. Eleven such sets were found to be essential for the production of biomass precursors. Experimental investigation of seven of these showed that knockouts of the associated genes resulted in attenuated growth for four pairs of reactions, whilst three single reactions were shown to be essential for growth.
Related JoVE Video
Molecular characterization of "inconsistent" variants of Salmonella Typhimurium isolated in Italy.
Foodborne Pathog. Dis.
PUBLISHED: 03-25-2014
Show Abstract
Hide Abstract
Salmonella 4,[5],12:i:- is a variant of Salmonella Typhimurium, which lacks the expression of phase-2 flagellar antigen, generally associated with the deletion of the fljB gene. Additional mechanisms involving the fljAB operon (?fljA, fljB, and hin genes) lead to the lack of expression of phase-2 flagellar antigens also in Salmonella strains harboring the fljB gene. For 20 S. 4,[5],12:i:- strains, defined as "inconsistent" Salmonella Typhimurium variants since they had phenotypically behaved as monophasic, even though the fljB gene was conserved, the fljAB operon was characterized in order to explain the ineffective expression of the phase-2 flagellar antigen. The monophasic phenotype for a first group of strains (9) was likely due to the absence of the hin gene, leading to the inhibited switch between the expression of phase-1 and phase-2 flagellar genes. For a second group of strains (5), the monophasic phenotype could be attributed to nonconservative point mutations identified in fljA and hin genes, which could hamper the proper expression of invertase gene and the fljA, acting as repressor of the phase-1 flagellar gene. Finally, for a last group of inconsistent strains (6), a plausible reason for their monophasic phenotype was not found, since the genes involved in the expression of phase-2 flagellar antigen were fully conserved. Moreover, the collection of inconsistent Salmonella Typhimurium isolates investigated were characterized by distinct molecular profiles, as demonstrated by multiple-locus variable-number tandem repeat analysis, and phenotype variability, as demonstrated by phage-typing. This study highlights the usefulness of investigating the entire fljAB operon when a definitive identification of the monophasic or biphasic status of Salmonella Typhimurium strains is needed (for instance, in the context of epidemiological investigations aimed to identify the relatedness among strains).
Related JoVE Video
Biocide and antibiotic susceptibility of Salmonella isolates obtained before and after cleaning at six Danish pig slaughterhouses.
Int. J. Food Microbiol.
PUBLISHED: 03-19-2014
Show Abstract
Hide Abstract
Salmonella sp. continues to be one of the most important foodborne pathogens. Control measures in terms of cleaning and disinfection on food production plants are very important for limiting the risk of contaminated food products to reach the consumer. In the last decade concern has arisen that bacteria exposed to disinfectants can develop resistance toward disinfectants and can have a higher risk of developing antibiotic resistance. The objectives of this study were to examine the prevalence of biocide resistant Salmonella sp. in Danish pig slaughterhouses, to evaluate if there was a correlation between susceptibilities to biocides and antibiotics, and to examine if cleaning and disinfection select isolates with changed susceptibility toward biocides or antibiotics. Salmonella sp. was isolated from the environment in Danish pig slaughterhouses before and after cleaning and disinfection. The susceptibility toward three different biocides, triclosan and two commercial disinfection products: Desinfect Maxi, a quaternary ammonium compound, and Incimaxx DES, an acetic compound, was determined. We found no resistance toward the biocides tested, but we did find that isolates obtained after cleaning had higher minimum inhibitory concentration (MIC) values toward one of the disinfectants (Incimaxx DES) compared to isolates obtained before cleaning and disinfection. This could indicate selection of strains that are more tolerant, due to the cleaning and disinfection. Furthermore, we found that there was a weak statistical correlation between MICs toward the biocides and some antibiotics, but no difference in log(MIC)s toward antibiotics between isolates obtained before and after cleaning, nor did we find any difference in the number of resistances of isolates obtained before and after cleaning and disinfection.
Related JoVE Video
Effects of environmental conditions on growth and survival of Salmonella in pasteurized whole egg.
Int. J. Food Microbiol.
PUBLISHED: 03-09-2014
Show Abstract
Hide Abstract
This study investigated the influence of three parameters (time, temperature and NaCl concentration) on survival and four parameters (temperature, NaCl and lysozyme concentrations and pH) on growth of Salmonella enterica serovar Enteritidis (S. Enteritidis) in pasteurized whole egg (PWE). Doehlert uniform shell design was employed to choose conditions for trials and data was fitted to polynomial models and were presented as estimated response surfaces. A model for prediction of reduction of S. Enteritidis in PWE within temperatures between 50 and 58°C, NaCl concentrations of 0-12%, and heating times between 30 and 210s and a model for prediction of growth rate of S. Enteritidis in PWE in the temperature range of 1-25°C, NaCl concentration of 0-12%, pH between 5 and 9, and lysozyme concentrations of 107-1007 U/mg proteins were developed. The maximum reduction condition was 58°C, 0% of NaCl at a fixed heating time of 120s, while maximum growth rate was estimated at 25°C and 0% of NaCl. pH and lysozyme concentration were shown not to influence growth performance significantly in the range of values studied. Results inform industry of the optimal pasteurization and storage parameters for liquid whole egg.
Related JoVE Video
European validation of a real-time PCR-based method for detection of Listeria monocytogenes in soft cheese.
Int. J. Food Microbiol.
PUBLISHED: 01-29-2014
Show Abstract
Hide Abstract
The classical microbiological method for detection of Listeria monocytogenes requires around 7 days for final confirmation, and due to perishable nature of RTE food products, there is a clear need for an alternative methodology for detection of this pathogen. This study presents an international (at European level) ISO 16140-based validation trial of a non-proprietary real-time PCR-based methodology that can generate final results in the following day of the analysis. This methodology is based on an ISO compatible enrichment coupled to a bacterial DNA extraction and a consolidated real-time PCR assay. Twelve laboratories from six European countries participated in this trial, and soft cheese was selected as food model since it can represent a difficult matrix for the bacterial DNA extraction and real-time PCR amplification. The limit of detection observed was down to 10 CFU per 25 of sample, showing excellent concordance and accordance values between samples and laboratories (>75%). In addition, excellent values were obtained for relative accuracy, specificity and sensitivity (82.75%, 96.70% and 97.62%, respectively) when the results obtained for the real-time PCR-based methods were compared to those of the ISO 11290-1 standard method. An interesting observation was that the L. monocytogenes detection by the real-time PCR method was less affected in the presence of Listeria innocua in the contaminated samples, proving therefore to be more reliable than the reference method. The results of this international trial demonstrate that the evaluated real-time PCR-based method represents an excellent alterative to the ISO standard since it shows a higher performance as well as reduce the extent of the analytical process, and can be easily implemented routinely by the competent authorities and food industry laboratories.
Related JoVE Video
The role of the st313-td gene in virulence of Salmonella Typhimurium ST313.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Multidrug-resistant Salmonella enterica serovar Typhimurium ST313 has emerged in sub-Saharan Africa causing severe infections in humans. Therefore, it has been speculated that this specific sequence type, ST313, carries factors associated with increased pathogenicity. We assessed the role in virulence of a gene with a yet unknown function, st313-td, detected in ST313 through comparative genomics. Additionally, the structure of the genomic island ST313-GI, harbouring the gene was determined. The gene st313-td was cloned into wild type S. Typhimurium 4/74 (4/74-C) as well as knocked out in S. Typhimurium ST313 02-03/002 (?st313-td) followed by complementation (02-03/002-C). ?st313-td was less virulent in mice following i.p. challenge than the wild type and this phenotype could be partly complemented in trans, indicating that st313-td plays a role during systemic infection. The gene st313-td was shown not to affect invasion of cultured epithelial cells, while the absence of the gene significantly affects uptake and intracellular survival within macrophages. The gene st313-td was proven to be strongly associated to invasiveness, harboured by 92.5% of S. Typhimurium blood isolates (n?=?82) and 100% of S. Dublin strains (n?=?50) analysed. On the contrary, S. Typhimurium isolates of animal and food origin (n?=?82) did not carry st313-td. Six human, non-blood isolates of S. Typhimurium from Belarus, China and Nepal harboured the gene and belonged to sequence types ST398 and ST19. Our data showed a global presence of the st313-td gene and in other sequence types than ST313. The gene st313-td was shown to be expressed during logarithmic phase of growth in 14 selected Salmonella strains carrying the gene. This study reveals that st313-td plays a role in S. Typhimurium ST313 pathogenesis and adds another chapter to understanding of the virulence of S. Typhimurium and in particular of the emerging sequence type ST313.
Related JoVE Video
Enumeration of Salmonella in table eggs, pasteurized egg products and egg containing dishes using quantitative real-time PCR.
Appl. Environ. Microbiol.
PUBLISHED: 12-20-2013
Show Abstract
Hide Abstract
Salmonellae are a major cause of food-borne outbreaks in Europe with eggs and egg products identified as major sources. Due to the low levels of Salmonella in eggs and egg products, direct quantification is difficult. In the present study enrichment quantitative real-time PCR (qPCR) was employed for enumeration of Salmonella in different matrices: table eggs, pasteurized egg products and egg containing dishes. Salmonella enterica serovar Enteritidis (S. Enteritidis) and S. Tennessee were used to artificially contaminate these matrices. Results showed a linear regression between the numbers of Salmonella and the quantification cycle (Cq) values for all matrices used with the exception of pasteurized egg white. Standard curves were constructed using both stationary phase cells and heat stressed cells with similar results. Finally the method was used to evaluate the fate of Salmonella in two egg containing dishes: long egg and tiramisu at abused refrigeration temperatures, and results indicated growth of bacteria over a week period. In conclusion enrichment qPCR was shown to be reliable for enumeration of Salmonella in different egg products.
Related JoVE Video
Molecular Characterization of Salmonella enterica serovar 4,[5],12:i:- DT193 ASSuT Strains from Two Outbreaks in Italy.
Foodborne Pathog. Dis.
PUBLISHED: 12-14-2013
Show Abstract
Hide Abstract
Abstract Salmonella enterica subsp. enterica serovar 4,[5],12:i:- DT193 is recognized as an emerging monophasic variant of Salmonella Typhimurium in many European countries. Resistance to ampicillin, streptomycin, sulphonamides, and tetracycline (R-type ASSuT) is described as one of the most common profiles of resistance within this clone. Recently, strains presenting such features were isolated from two unrelated outbreaks in Italy. Strains were characterized by pulsed-field gel electrophoresis (PFGE), performed with XbaI, BlnI, and SpeI, and multiple-locus variable-number tandem repeat analysis (MLVA). XbaI-PFGE showed strains related to the two outbreaks as indistinguishable. Conversely, both BlnI-PFGE and MLVA characterized the strains related the two outbreaks as different. XbaI-PFGE identified two profiles, differing by one band, within strains isolated from one of the two outbreaks. Also BlnI-PFGE and MLVA generated different profiles among the strains related to that outbreak. Combining the PFGE profiles obtained by XbaI and BlnI and comparing them with the MLVA profiles, the two methods grouped the same isolates based on identity. Moreover, genomic deletions of the genes included in the operon fljAB, the flanking iroB gene, and the closely located STM2757 gene were investigated. For all strains, the same profile of deletion characterized by the absence of fljA, fljB, and hin genes and the presence of STM2757 and iroB genes was identified. This profile of deletion represents a mixture between two profiles of Salmonella 4,[5],12:i:- described as the "Spanish" and the "U.S." clones. This study demonstrated that although strains of Salmonella 4,[5],12:i:- DT193 ASSuT are highly clonal, minor differences between strains may be seen during the same outbreak by using in parallel PFGE with different restriction enzymes, MLVA, and the analysis of molecular markers related to the operon fljAB. The combination of these different molecular approaches was essential to clarify the epidemiological relationship among the strains.
Related JoVE Video
The role of flagella and chemotaxis genes in host pathogen interaction of the host adapted Salmonella enterica serovar Dublin compared to the broad host range serovar S. Typhimurium.
BMC Microbiol.
PUBLISHED: 03-19-2013
Show Abstract
Hide Abstract
The importance of flagella and chemotaxis genes in host pathogen interaction in Salmonella enterica is mainly based on studies of the broad host range serovar, S. Typhimurium, while little is known on the importance in host specific and host adapted serovars, such as S. Dublin. In the current study we have used previously characterized insertion mutants in flagella and chemotaxis genes to investigate this and possible differences in the importance between the two serovars.
Related JoVE Video
Intestinal invasion of Salmonella enterica serovar Typhimurium in the avian host is dose dependent and does not depend on motility and chemotaxis.
Vet. Microbiol.
PUBLISHED: 01-25-2013
Show Abstract
Hide Abstract
Salmonella enterica serotype Typhimurium (S. Typhimurium) can invade in the intestine of the avian host, and knowledge on the mechanisms that govern this is potentially important for prevention of disease. This study investigated the invasion of S. Typhimurium in the avian host and to which extent it depended on motility and chemotaxis. Wild type and previously well-characterized transposon mutants in flagella genes fliC and fljB and in chemotaxis genes cheA, cheB and cheR were used as challenge strains in intestinal loop experiments. Invasion was shown to be dose dependent, but did not require functional flagella or chemotaxis genes. In support of the results from intestinal loop experiments, flagella and chemotaxis genes were not significantly important to the outcome of an oral infection. The results showed that S. Typhimurium invasion in the avian host was dose dependent and was not affected by the loss of flagella and chemotaxis genes.
Related JoVE Video
Salmonella source attribution based on microbial subtyping.
Int. J. Food Microbiol.
PUBLISHED: 01-20-2013
Show Abstract
Hide Abstract
Source attribution of cases of food-borne disease represents a valuable tool for identifying and prioritizing effective food-safety interventions. Microbial subtyping is one of the most common methods to infer potential sources of human food-borne infections. So far, Salmonella microbial subtyping source attribution models have been implemented by using serotyping and phage-typing data. Molecular-based methods may prove to be similarly valuable in the future, as already demonstrated for other food-borne pathogens like Campylobacter. This review assesses the state of the art concerning Salmonella source attribution through microbial subtyping approach. It summarizes the available microbial subtyping attribution models and discusses the use of conventional phenotypic typing methods, as well as of the most commonly applied molecular typing methods in the European Union (EU) laboratories in the context of their potential applicability for Salmonella source attribution studies.
Related JoVE Video
Evidence of broiler meat contamination with post-disinfection strains of Campylobacter jejuni from slaughterhouse.
Int. J. Food Microbiol.
PUBLISHED: 04-06-2010
Show Abstract
Hide Abstract
While cross-contamination from equipment and scalding water containing Campylobacter jejuni is considered the main route of broiler carcass contamination during slaughtering, alternative sources of C. jejuni may have been overlooked because only a limited number of studies focus on sampling of one broiler flock along the entire food chain and not many include the slaughterhouse environment. In the present study we have traced the changes of C. jejuni genotypes within one broiler flock from the beginning of rearing to the final product at the slaughterhouse with the aim to evaluate the dynamics and possible sources of carcass contamination with C. jejuni. Genotyping of 345 isolates of C. jejuni by flaA-RFLP revealed ten different flaA genotypes of C. jejuni along the broiler meat production chain. Broiler fillets were mainly contaminated with flaA genotypes found on the surfaces of slaughterhouse equipment and in the scalding water after cleaning and disinfection. Finally, it was clearly demonstrated that C. jejuni isolates remaining in the slaughterhouse environment after disinfection is a potential source of broiler meat contamination. Thus, identification of the mechanisms that allow such strains to persist in the slaughterhouse and survive cleaning is important for the establishment of future practices that will ensure sufficient reduction of C. jejuni in the slaughterhouse environment.
Related JoVE Video
The in vitro fitness cost of antimicrobial resistance in Escherichia coli varies with the growth conditions.
FEMS Microbiol. Lett.
PUBLISHED: 07-22-2009
Show Abstract
Hide Abstract
The objective of this study was to investigate the influence of stressful growth conditions on the fitness cost of antimicrobial resistance in Escherichia coli BJ4 caused by chromosomal mutations and plasmid acquisition. The fitness cost of chromosomal streptomycin resistance increased significantly when the bacteria were grown under all stress conditions tested, while the cost in 1/3 Luria-Bertani was not significantly changed in a streptomycin+rifampicin mutant. The increase in the fitness cost depended in a nonregular manner on the strain/stress combination. The fitness cost of plasmid-encoded resistance on R751 did not differ significantly, and was generally less under stressful growth conditions than in rich media. The fitness cost associated with R751 with the multiple drug resistance cassette from Salmonella Typhimurium DT104 increased significantly only under stressful conditions at low pH and at high-salt concentrations. Strains with an impaired rpoS demonstrated a reduced fitness only during growth in a high-salt concentration. In conclusion, it was demonstrated that bacterial fitness cost in association with antimicrobial resistance generally increases under stressful growth conditions. However, the growth potential of bacteria with antimicrobial resistances did not increase in a straightforward manner in these in vitro experiments and is therefore probably even more difficult to predict in vivo.
Related JoVE Video
Polyamines are required for virulence in Salmonella enterica serovar Typhimurium.
PLoS ONE
Show Abstract
Hide Abstract
Sensing and responding to environmental cues is a fundamental characteristic of bacterial physiology and virulence. Here we identify polyamines as novel environmental signals essential for virulence of Salmonella enterica serovar Typhimurium, a major intracellular pathogen and a model organism for studying typhoid fever. Central to its virulence are two major virulence loci Salmonella Pathogenicity Island 1 and 2 (SPI1 and SPI2). SPI1 promotes invasion of epithelial cells, whereas SPI2 enables S. Typhimurium to survive and proliferate within specialized compartments inside host cells. In this study, we show that an S. Typhimurium polyamine mutant is defective for invasion, intracellular survival, killing of the nematode Caenorhabditis elegans and systemic infection of the mouse model of typhoid fever. Virulence of the mutant could be restored by genetic complementation, and invasion and intracellular survival could, as well, be complemented by the addition of exogenous putrescine and spermidine to the bacterial cultures prior to infection. Interestingly, intracellular survival of the polyamine mutant was significantly enhanced above the wild type level by the addition of exogenous putrescine and spermidine to the bacterial cultures prior to infection, indicating that these polyamines function as an environmental signal that primes S. Typhimurium for intracellular survival. Accordingly, experiments addressed at elucidating the roles of these polyamines in infection revealed that expression of genes from both of the major virulence loci SPI1 and SPI2 responded to exogenous polyamines and was reduced in the polyamine mutant. Together our data demonstrate that putrescine and spermidine play a critical role in controlling virulence in S. Typhimurium most likely through stimulation of expression of essential virulence loci. Moreover, our data implicate these polyamines as key signals in S. Typhimurium virulence.
Related JoVE Video
Mustelidae are natural hosts of Staphylococcus delphini group A.
Vet. Microbiol.
Show Abstract
Hide Abstract
According to the current taxonomy, the Staphylococcus intermedius group (SIG) comprises of at least three distinct species. While S. intermedius and S. pseudintermedius are associated with specific hosts (pigeons and dogs, respectively), the natural host of S. delphini remains unclear. We analysed 158 SIG isolates from less studied animal species belonging to the order Carnivora, including mink (n=118), fox (n=33), badger (n=6) and ferret (n=1). Species identification was performed by nuc PCR in combination with sodA sequence analysis and pta PCR restriction fragment length polymorphism (RFLP). The results showed a consistent association between host and bacterial species. All isolates from minks, ferret and badgers belonged to S. delphini group A, whereas all fox isolates except one were identified as S. pseudintermedius. The remaining fox isolate belonged to S. delphini group A. The results indicate that Mustelidae such as minks, ferrets and badgers are natural hosts of S. delphini group A. This is in contrast with Canidae, which are primarily colonized and infected with S. pseudintermedius. These findings suggest that coagulase-positive staphylococcal species may have evolved and diverged through host adaptation.
Related JoVE Video

What is Visualize?

JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

How does it work?

We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

Video X seems to be unrelated to Abstract Y...

In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.