Balance between bacterial extracellular matrix production and intramacrophage proliferation by a Salmonella-specific SPI-2-encoded transcription factor.

Biosynthesis and secretion of a complex extracellular matrix (EM) is a hallmark of Salmonella biofilm formation, impacting on its relationship with both the environment and the host. Cellulose is a major component of Salmonella EM. It is considered an anti-virulence factor because it interferes with Salmonella proliferation inside macrophages and virulence in mice. Its synthesis is stimulated by CsgD, the master regulator of biofilm formation in enterobacteria, which in turn is under the control of MlrA, a MerR-like transcription factor. In this work, we identified a SPI-2-encoded Salmonella-specific transcription factor homolog to MlrA, MlrB, that represses transcription of its downstream gene, orf319, and of csgD inside host cells. MlrB is induced in laboratory media mimicking intracellular conditions and inside macrophages, and it is required for intramacrophage proliferation. An increased csgD expression is observed in the absence of MlrB inside host cells. Interestingly, inactivation of the CsgD-controlled cellulose synthase-coding gene restored intramacrophage proliferation to rates comparable to wild-type bacteria in the absence of MlrB. These data indicate that MlrB represses CsgD expression inside host cells and suggest that this repression lowers the activation of the cellulose synthase. Our findings provide a novel link between biofilm formation and Salmonella virulence.

Salmonella Typhimurium ST313 isolated in Brazil revealed to be more invasive and inflammatory in murine colon compared to ST19 strains.

Salmonella Typhimurium (ST313) has caused an epidemic of invasive disease in sub-Saharan Africa and has been recently identified in Brazil. As the virulence of this ST is poorly understood, the present study aimed to (i) perform the RNA-seq in vitro of S. Typhimurium STm30 (ST313) grown in Luria-Bertani medium at 37°C; (ii) compare it with the RNA-seq of the S. Typhimurium SL1344 (ST19) and S. Typhimurium STm11 (ST19) strains under the same growing conditions; and (iii) examine the colonization capacity and expression of virulence genes and cytokines in murine colon. The STm30 (ST313) strain exhibited stronger virulence and was associated with a more inflammatory profile than the strains SL1344 (ST19) and STm11 (ST19), as demonstrated by transcriptome and in vivo assay. The expression levels of the hilA, sopD2, pipB, and ssaS virulence genes, other Salmonella pathogenicity islands SPI-1 and SPI-2 genes or effectors, and genes of the cytokines IL-1ß, IFN-γ, TNF-α, IL-6, IL-17, IL-22, and IL-12 were increased during ST313 infection in C57BL/6J mice. In conclusion, S. Typhimurium STm30 (ST313) isolated from human feces in Brazil express higher levels of pathogenesis-related genes at 37°C and has stronger colonization and invasion capacity in murine colon due to its high expression levels of virulence genes, when compared with the S. Typhimurium SL1344 (ST19) and STm11 (ST19) strains. STm30 (ST313) also induces stronger expression of pro-inflammatory cytokines in this organ, suggesting that it causes more extensive tissue damage.

Phylogenetic relationship and genomic characterization of Salmonella Typhimurium strains isolated from swine in Brazil.

Salmonella Typhimurium has been transmitted between humans and animals. Although, Brazil has been one of the largest pork meat exporters worldwide, there are few studies that characterized epidemiologically S. Typhimurium strains from swine. The aims of this work were to study the phylogenetic relationship of S. Typhimurium genomes isolated from swine in Brazil among themselves and with other genomes isolated from several sources and countries using wgMLST and cgMLST and to perform the search of Salmonella pathogenicity islands (SPIs). In addition, for S. Typhimurium strains from swine to compare the virulence and antimicrobial resistance genes by VFDB and ResFinder, genetic content by BLAST Atlas and orthologous proteins clusters by OrthoVenn. The constructed phylogenetic trees by wgMLST and cgMLST grouped the majority (92.3% and 80.7%, respectively) of the strains isolated from swine in Brazil into the same group. All the isolates contained important SPIs (SPI-1, SPI-2, SPI-3, SPI-5 and SPI-9). A total of 100 and 31 virulence and resistance genes were detected in the S. Typhimurium strains isolated from swine, respectively. The BLAST Atlas and orthologous proteins analysis found regions of phages and differences in metabolic, regulatory and cellular processes among S. Typhimurium LT2 and S. Typhimurium isolates from swine. In conclusion, molecular typing based in the wgMLST and cgMLST suggested that the S. Typhimurium isolates from swine studied were genetically related. The pathogenic potential of the strains studied was corroborated by the presence of important SPIs and virulence genes. The high number of antimicrobial resistance genes detected is worrying and reinforced their potential risk in swine in Brazil. The comparison by BLAST Atlas suggested differences in mobile genetic elements among S. Typhimurium LT2 and S. Typhimurium isolates from swine in Brazil. The orthologous proteins analysis revealed unique genes related to important cellular processes in the strains from swine.

Perspectives for the use of latex peptidases from Calotropis procera for control of inflammation derived from Salmonella infections.

BACKGROUND: Anti-inflammatory properties have been attributed to latex proteins of the medicinal plant Calotropis procera. PURPOSE: A mixture of cysteine peptidases (LPp2) from C. procera latex was investigated for control of inflammatory mediators and inflammation in a mouse model of Salmonella infection. METHODS: LPp2 peptidase activity was confirmed by the BANA assay. Cytotoxicity assays were conducted with immortalized macrophages. Peritoneal macrophages (pMØ) from Swiss mice were stimulated with lipopolysaccharide (LPS) in 96-well plates and then cultured with nontoxic concentrations of LPp2. Swiss mice intravenously received LPp2 (10 mg/kg) and then were challenged intraperitoneally with virulent Salmonella enterica Ser. Typhimurium. RESULTS: LPp2 was not toxic at dosages lower than 62.2 µg/mL. LPp2 treatments of pMØ stimulated with LPS impaired mRNA expression of pro-inflammatory cytokines IL-1ß, TNF-α, IL-6 and IL-10. LPp2 increased the intracellular bacterial killing in infected pMØ. Mice given LPp2 had a lower number of leukocytes in the peritoneal cavity in comparison to control groups 6 h after infection. The bacterial burden and histological damage were widespread in target organs of mice receiving LPp2. CONCLUSION: We conclude that LPp2 contains peptidases with strong anti-inflammatory properties, which may render mice more susceptible to early disseminated infection caused by Salmonella.

A dynamic and integrated in vitro/ex vivo gastrointestinal model for the evaluation of the probability and severity of infection in humans by Salmonella spp. vehiculated in different matrices.

The lack of proper gastrointestinal models assessing the inter-strain virulence variability of foodborne pathogens and the effect of the vehicle (food matrix) affects the risk estimation. This research aimed to propose a dynamic and integrated in vitro/ex vivo gastrointestinal model to evaluate the probability and severity of infection of foodborne pathogens at different matrices. An everted gut sac was used to determine the adhesion and invasion of Salmonella enterica and tissue damage. S. Typhimurium ATCC 14028 was used as a representative bacterium, and two matrices (water and cheese) were used as vehicles. No differences (p > 0.05) in the probability of infection (Pinf) were found for intra-experimental repeatability. However, the Pinf of cheese-vehiculated S. Typhimurium was different compared to water- vehiculated S. Typhimurium, 7.2-fold higher. The histological analysis revealed Salmonella-induced tissue damage, compared with the control (p < 0.05). In silico proposed interactions between two major Salmonella outer membrane proteins (OmpA and Rck) and digested peptides from cheese casein showed high binding affinity and stability, suggesting a potential protective function from the food matrix. The results showed that the everted gut sac model is suitable to evaluate the inter-strain virulence variability, considering both physiological conditions and the effect of the food matrix.

Avaliação in vitro de atividade antimicrobiana de óleos essenciais contra Salmonella typhimurium e Staphylococcus aureus / In vitro evaluation of antimicrobial activity of essential oils against Salmonella typhimurium and Staphylococcus aureus

O objetivo do trabalho foi avaliar a atividade antimicrobiana in vitro de diferentes óleos essenciais e suas combinações contra linhagens patogênicas de Salmonella typhimurium ATCC 13311 e Staphylococcus aureus ATCC 13565. Foram testados ao todo 41 óleos essenciais e as combinações em pares dos óleos de alecrim-pimenta (Lippia sidoides Cham.), capim-limão (Cymbopogon citratus Stapf.), tomilho (Thymus vulgaris L.), orégano (Origanum vulgare L.) e canela (Cinnamomum cassia (L.) D.Don. para observar efeitos adicionais ou não na inibição dos microrganismos. Os resultados mostraram que as melhores atividades antimicrobianas para os dois microrganismos foram obtidas utilizando os óleos de Thymus vulgaris L. (tomilho), Origanum vulgare L. (orégano) e Lippia sidoides Cham. (alecrim-pimenta). As combinações de cada par dos óleos de alecrim-pimenta, capim-limão, tomilho, orégano e canela não apresentaram uma melhora adicional no efeito para os patógenos estudados quando comparados com os seus óleos testados isoladamente. As composições químicas dos óleos essenciais mais ativos foram obtidas por meio de análises por cromatografia gasosa acoplada à espectrometria de massas (CG-EM), sendo que os compostos majoritários do óleo de orégano foram carvacrol (69,1%) e p-cimeno (18,8%), do óleo essencial de tomilho foram timol (45,5%) e p-cimeno (35,6%) e do alecrim-pimenta foram o timol (77,2%) e p-cimeno (14,2%). Análises quantitativas de p-cimeno, carvacrol e timol nos três óleos foram realizadas e comparadas com as composições químicas mostrando resultados proporcionalmente coerentes, exceto para p-cimeno, que foi quantificado com menores porcentagens por esse método. A aplicabilidade dos óleos selecionados, ou das substâncias presentes isoladas seria diretamente nos alimentos, com a finalidade de inibir ou controlar o crescimento desses patógenos, contribuindo assim para a segurança dos alimentos. (AU)

The aim of this study was to evaluate the in vitro antimicrobial activity of several essential oils and their combination against pathogenic strains of Salmonella typhimurium ATCC 13311 and Staphylococcus aureus ATCC 13565. A total of 41 essential oils and the combinations in pairs of the oils of rosemary pepper (Lippia sidoides Cham.), lemongrass (Cymbopogon citratus Stapf.), thyme (Thymus vulgaris L.), origanum (Origanum vulgare L.) and cinnamon (Cinnamomum cassia (L.) D.Don) were tested to observe additional effects or not in the inhibition of microorganisms. The results showed that the best antimicrobial activities for the two microorganisms were obtained using the oils of Thymus vulgaris L. (thyme), Origanum vulgare L. (origanum) and Lippia sidoides Cham. (rosemary pepper). The combinations of each pair of rosemary pepper, lemongrass, thyme, origanum and cinnamon oils did not show any further improvement in the effect for the studied pathogens when compared to their oils tested alone. The compositions of the most active essential oils were obtained by gas chromatography coupled to mass spectrometry (GC-MS). The major compounds in origanum oil were carvacrol (69.1%) and p-cymene (18.8%), thyme essential oil were thymol (45.5%) and p-cymene (35.6%) and rosemary pepper were thymol (77.2%) and p-cymene (14.2%). Quantitative analyses of p-cymene, carvacrol and thymol confirmed the presence in thyme, oregano and pepper rosemary oils and were compared with the chemical composition showing similar results, except for p-cymene, which was quantified with lower percentages in these oils. One of the possible applications of the results of this work is the use of essential oils, or the isolated substances, directly in foods, with the purpose of inhibiting or controlling the growth of these pathogens, thus contributing to the food safety.Keywords: Bioassay. GC-MS. Inhibition. Carvacrol. Thymol. p-cymene(AU)

Class 1 integron-borne cassettes harboring blaCARB-2 gene in multidrug-resistant and virulent Salmonella Typhimurium ST19 strains recovered from clinical human stool samples, United States.

International lineages, such as Salmonella Typhimurium sequence type (ST) 19, are most often associated with foodborne diseases and deaths in humans. In this study, we compared the whole-genome sequences of five S. Typhimurium strains belonging to ST19 recovered from clinical human stool samples in North Carolina, United States. Overall, S. Typhimurium strains displayed multidrug-resistant profile, being resistance to critically and highly important antimicrobials including ampicillin, ticarcillin/clavulanic acid, streptomycin and sulfisoxazole, chloramphenicol, tetracycline, respectively. Interestingly, all S. Typhimurium strains carried class 1 integron (intl1) and we were able to describe two genomic regions surrounding blaCARB-2 gene, size 4,062 bp and 4,422 bp for S. Typhimurium strains (HS5344, HS5437, and HS5478) and (HS5302 and HS5368), respectively. Genomic analysis for antimicrobial resistome confirmed the presence of clinically important genes, including blaCARB-2, aac(6')-Iaa, aadA2b, sul1, tetG, floR, and biocide resistance genes (qacEΔ1). S. Typhimurium strains harbored IncFIB plasmid containing spvRABCD operon, as well as rck and pef virulence genes, which constitute an important apparatus for spreading the virulence plasmid. In addition, we identified several virulence genes, chromosomally located, while the phylogenetic analysis revealed clonal relatedness among these strains with S. enterica isolated from human and non-human sources obtained in European and Asian countries. Our results provide new insights into this unusual class 1 integron in virulent S. Typhimurium strains that harbors a pool of genes acting as potential hotspots for horizontal gene transfer providing readily adaptation to new surrounds, as well as being crucially required for virulence in vivo. Therefore, continuous genomic surveillance is an important tool for safeguarding human health.

Phenotypic and genotypic characterization of Salmonella Typhimurium isolates from humans and foods in Brazil.

Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) causes gastroenteritis in many countries. However, in Brazil there are few studies that have conducted a virulence characterization of this serovar. The aim of this study was to evaluate the virulence potential of S. Typhimurium strains isolated in Brazil. Forty S. Typhimurium strains isolated from humans (n = 20) and food (n = 20) from Brazil were studied regarding their invasion and survival in human epithelial cells (Caco-2) and macrophages (U937). Their virulence potential was determined using the Galleria mellonella larvae model combined with the analysis of virulence genes by whole genome sequencing (WGS). A total of 67.5% of the S. Typhimurium studied (32.5% isolated from humans and 35% isolated from food) invaded Caco-2 epithelial cells at levels similar to or greater than the S. Typhimurium SL1344 prototype strain. In addition, 37.5% of the studied strains (25% isolated from humans and 12.5% isolated from food) survived in U937 human macrophages at levels similar to or greater than SL1344. S. Typhimurium strains isolated from humans (40%) and food (25%) showed high or intermediate virulence in G. mellonella larvae after seven days exposure. Approximately, 153 virulence genes of chromosomal and plasmidial origin were detected in the strains studied. In conclusion, the ability of the S. Typhimurium to invade Caco-2 epithelial cells was strain dependent and was not related to the source or the year of isolation. However, S. Typhimurium strains isolated from humans showed greater survival rates in U937 human macrophages, and presented higher proportion of isolates with a virulent profile in G. mellonella in comparison to strains isolated from food suggesting that this difference may be related to the higher frequency of human isolates which contained plasmid genes, such as spvABCDR operon, pefABCD operon, rck and mig-5.

PhoQ is an unsaturated fatty acid receptor that fine-tunes Salmonella pathogenic traits.

The Salmonella enterica PhoP/PhoQ two-component signaling system coordinates the spatiotemporal expression of key virulence factors that confer pathogenic traits. Through biochemical and structural analyses, we found that the sensor histidine kinase PhoQ acted as a receptor for long-chain unsaturated fatty acids (LCUFAs), which induced a conformational change in the periplasmic domain of the PhoQ protein. This resulted in the repression of PhoQ autokinase activity, leading to inhibition of the expression of PhoP/PhoQ-dependent genes. Recognition of the LCUFA linoleic acid (LA) by PhoQ was not stereospecific because positional and geometrical isomers of LA equally inhibited PhoQ autophosphorylation, which was conserved in multiple S. enterica serovars. Because orally acquired Salmonella encounters conjugated LA (CLA), a product of the metabolic conversion of LA by microbiota, in the human intestine, we tested how short-term oral administration of CLA affected gut colonization and systemic dissemination in a mouse model of Salmonella-induced colitis. Compared to untreated mice, CLA-treated mice showed increased gut colonization by wild-type Salmonella, as well as increased dissemination to the spleen. In contrast, the inability of the phoP strain to disseminate systemically remained unchanged by CLA treatment. Together, our results reveal that, by inhibiting PhoQ, environmental LCUFAs fine-tune the fate of Salmonella during infection. These findings may aid in the design of new anti-Salmonella therapies.

Type three secretion system in Salmonella Typhimurium: the key to infection.

BACKGROUND: Type Three Secretion Systems (T3SS) are nanomachine complexes, which display the ability to inject effector proteins directly into host cells. This skill allows for gram-negative bacteria to modulate several host cell responses, such as cytoskeleton rearrangement, signal transduction, and cytokine production, which in turn increase the pathogenicity of these bacteria. The Salmonella enterica subsp. enterica serovar Typhimurium (ST) T3SS has been the most characterized so far. Among gram-negative bacterium, ST is one of enterica groups predicted to have two T3SSs activated during different phases of infection. OBJECTIVE: To comprise current information about ST T3SS structure and function as well as an overview of its assembly and hierarchical regulation. METHODS: With a brief and straightforward reading, this review summarized aspects of both ST T3SS, such as its structure and function. That was possible due to the development of novel techniques, such as X-ray crystallography, cryoelectron microscopy, and nano-gold labelling, which also elucidated the mechanisms behind T3SS assembly and regulation, which was addressed in this review. CONCLUSION: This paper provided fundamental overview of ST T3SS assembly and regulation, besides summarized the structure and function of this complex. Due to T3SS relevance in ST pathogenicity, this complex could become a potential target in therapeutic studies as this nanomachine modulates the infection process.

Milk Fermented by Lactobacillus paracasei NCC 2461 (ST11) Modulates the Immune Response and Microbiota to Exert its Protective Effects Against Salmonella typhimurium Infection in Mice.

Probiotics form a promising strategy to maintain intestinal health. Milks fermented with probiotic strains, such as the Lactobacillus paracasei ST11, are largely commercialized in Brazil and form a low-cost alternative to probiotic pharmaceutical formulations. In this study, we assessed the probiotic effects of milk fermented by L. paracasei ST11 (administered through fermented milk) in a Salmonella typhimurium infection model in BALB/c mice. We observed in this murine model that the applied probiotic conferred protective effects against S. typhimurium infection, since its administration reduced mortality, weight loss, translocation to target organs (liver and spleen) and ileum injury. Moreover, a reduction in the mRNA expression of pro-inflammatory cytokines such as IFN-γ, IL-6, TNF-α and IL-17 in animals that received the probiotic before challenge was observed. Additionally, the ileum microbiota was better preserved in these animals. The present study highlights a multifactorial protective aspect of this commercial probiotic strain against a common gastrointestinal pathogen.

Stability of the Salmonella Typhimurium rcsC11 mutant under different stress conditions.

The virulence genes of Salmonella are modulated during infection by several regulatory systems, and the RcsCDB system is one of the most important of these. The S. Typhimurium EG14873 (rcsC11) strain harbours the rcsC11 point mutation, displaying a constitutive activation of this system, which is characterized by mucoid colonies and attenuated virulence phenotypes. In this work, the stability of the rcsC11 mutation was analysed under stress conditions. Under acid and anaerobic stresses, we observed the appearance of small and non-mucoid colonies of the rcsC11 strain. The sequencing of the rcsC gene from these colonies showed that the mutation is conserved. Moreover, we found that small colonies were also generated when the wild-type strain grew in acid and anaerobic conditions. It is worth noting that the transition from normal to atypical colonies of both strains only took place after several days of incubation and was not observed during eukaryotic cell infection. Therefore, the appearance of these atypical colonies is a characteristic feature of S. Typhimurium strains under stressful situations and does not involve a reversion of the rcsC11 allele and nor does it imply any risk to mammalian cells. Therefore, we propose that the S. Typhimurium rcsC11 strain is a good candidate for the development of attenuated vaccines.

Assessment and incidence of fish associated bacterial pathogens at hatcheries of Azad Kashmir, Pakistan / Avaliação e incidência de patógenos bacterianos associados a peixes em incubadoras da Caxemira de Azad, Paquistão

Fish is the most indispensable source of proteins for individuals and have high nutritional value. On the other hand, the fish culturing raised issues of fish health due to close contact between the aquatic environment and the fish pathogens. So, the aim of the current study was to identify the bacterial pathogens and screen the injured Rainbow trout rearing in different trout hatcheries run under fisheries department of the government of Azad Jammu and Kashmir, Pakistan. Seven bacterial pathogens such as Shigella flexneri, Enterobacter amnigenus, Salmonella Typhimurium, Serratia odorifera, Pseudomonas aeruginosa, Streptococcus pyogenes, and Bacillus cereus were isolated and identified. Results revealed that the injury of fish specimens was due to overcrowding. Instead of rainbow coloration, specimens have darker black in color. The water of ponds was not clean and clear and such conditions was because of the greater quantity of feed thrown in the water. It was concluded that poor hygienic water condition and overloading allowed the opportunistic bacterial contaminations to succeed which cause a serious threat to hatcheries.(AU)

O peixe é a fonte mais indispensável de proteínas para os indivíduos e tem alto valor nutricional. Por outro lado, a cultura dos peixes levantou questões sobre a saúde dos peixes devido ao próximo contato entre o ambiente aquático e os agentes patogênicos desses peixes. Assim, o objetivo do presente estudo foi identificar os patógenos bacterianos e rastrear a criação da truta arco-íris que apresentou lesões em diferentes incubadoras de trutas, com supervisão do departamento de pesca do governo de Azad Jammu e Caxemira, Paquistão. Sete patógenos bacterianos foram isolados e identificados, tais como: Shigella flexneri, Enterobacter amnigenus, Salmonella typhimurium, Serratia odorifera, Pseudomonas aeruginosa, Streptococcus pyogenes e Bacillus cereus. Os resultados revelaram que a lesão de espécimes de peixes foi devido à superlotação. Em vez da coloração do arco-íris, os espécimes tiveram uma coloração preta mais escura. A água das lagoas não era limpa e nem clara, e tais condições ocorreram devido a maior quantidade de alimento lançada na água. Concluiu-se que a precária condição higiênica da água e também a sobrecarga permitiram que as contaminações bacterianas oportunistas fossem bem-sucedidas, causando séria ameaça às incubadoras.(AU)

Population analysis of D6-like plasmid prophage variants associated with specific IncC plasmid types in the emerging Salmonella Typhimurium ST213 genotype.

The Salmonella enterica serovar Typhimurium sequence type 213 (ST213) emerged as a predominant genotype in Mexico. It is characterized by harboring multidrug resistance (MDR) IncC plasmids (previously IncA/C) and the lack of the Salmonella virulence plasmid (pSTV). Here we show that the D6-like plasmid prophage is present in most of the ST213 strains. We used the reported nucleotide sequence of YU39 plasmid (pYU39_89) to design a PCR typing scheme for the D6-like plasmid prophages, and determined the complete nucleotide sequences for the D6-like prophages of three additional ST213 strains (YU07-18, SL26 and SO21). Two prophage variants were described: i) a complete prophage, containing homologous sequences for most of the genetic modules described in P1 and D6 phages, which most likely allow for the lytic and lysogenic lifestyles; and ii) an incomplete prophage, lacking a 15 kb region containing morphogenesis genes, suggesting that it is defective. The tail fiber gene inversion region was the most divergent one between D6 and pYU39_89 genomes, suggesting the production of a distinct set of tail fibers, which could be involved in host range preferences. A glutaminyl-tRNA synthetase gene (glnS), which could be involved in providing host cell increased fitness or plasmid maintenance functions, was found in all D6-like genomes. Population level analysis revealed a biogeographic pattern of distribution of these plasmid-phages and specific associations with variants of MDR IncC plasmids. Statistically significant associations were found between the two prophage variants (p75 or p89), the type of IncC plasmids (I or II) and geographic isolation regions (Sonora, San Luis Potosí, Michoacán and Yucatán). This work integrates results from molecular typing, genomics and epidemiology to provide a broad overview for the evolution of an emergent Salmonella genotype.

Influência do ultrassom no crescimento de Salmonella typhimurium / Influence of ultrasound in Salmonella typhimurium growth

O ultrassom tem sido amplamente estudado na inativação de microrganismos e no processamento de alimentos. Entretanto, o efeito sobre o crescimento de bactérias ainda é pouco elucidado. O presente estudo avaliou o efeito de diferentes densidades energéticas de ultrassom (0; 0,11; 0,22 e 0,43 KJ/mL) no crescimento de Salmonella enterica serovar Typhimurium em caldo BHI. Altas densidades energéticas diminuíram (p < 0,05) a duração da fase lag e a densidade de células na fase estacionária. Entretanto, não houve diferença (p < 0,05) na velocidade de crescimento. Portanto, os resultados do presente estudo alertam para os riscos do ultrassom quando aplicado isoladamente. Assim, as aplicações desta tecnologia, isoladamente, em alimentos que suportam o crescimento de patógenos, como carne e produtos cárneos, devem ser minuciosamente avaliadas para cada tipo de produto.(AU)

Análise epidemiológica da distribuição dos genes de resistência antimicrobiana de Salmonella typhimurium no Brasil / Epidemiological analysis of the antimicrobial resistance genes distribution of Salmonella typhimurium in Brazil

Salmonella é uma das principais enterobactérias causadoras de gastroenterites no Brasil e no mundo, sendo Typhimurium o principal sorotipo. A disseminação desta bactéria causa perdas econômicas e produtivas. Nos países em desenvolvimento ocorre subnotificação dos surtos de salmoneloses prejudicando a vigilância epidemiológica. O uso de antibióticos é o tratamento mais eficiente contra infecções por Salmonella. Porém, seu uso indiscriminado favorece a seleção de linhagens resistentes e dificulta o tratamento das salmoneloses. Foi realizado um estudo da distribuição epidemiológica dos genes de resistência antimicrobiana adquirida em estirpes de Salmonella Typhimurium isoladas no Brasil, no período de 1983 a 2013. A classe de antibióticos que apresentou maior quantidade de genes de resistência foi a dos aminoglicosídeos, sendo o gene com maior prevalência o aac(6')-Iaa. As estirpes estudadas apresentaram resistência a antibióticos pertencentes entre 1 a 7 classes diferentes.

Influência do ultrassom no crescimento de Salmonella typhimurium / Influence of ultrasound in Salmonella typhimurium growth

O ultrassom tem sido amplamente estudado na inativação de microrganismos e no processamento de alimentos. Entretanto, o efeito sobre o crescimento de bactérias ainda é pouco elucidado. O presente estudo avaliou o efeito de diferentes densidades energéticas de ultrassom (0; 0,11; 0,22 e 0,43 KJ/mL) no crescimento de Salmonella enterica serovar Typhimurium em caldo BHI. Altas densidades energéticas diminuíram (p < 0,05) a duração da fase lag e a densidade de células na fase estacionária. Entretanto, não houve diferença (p < 0,05) na velocidade de crescimento. Portanto, os resultados do presente estudo alertam para os riscos do ultrassom quando aplicado isoladamente. Assim, as aplicações desta tecnologia, isoladamente, em alimentos que suportam o crescimento de patógenos, como carne e produtos cárneos, devem ser minuciosamente avaliadas para cada tipo de produto.

Patogenia de mutantes de Salmonella Typhimurium en dos modelos experimentales in vivo / Pathogenesis of Salmonella Typhimurium mutants in two experimental models in vivo

Con la finalidad de evaluar la patogenia en cepas de Salmonella Typhimurium con mutaciones en los genes invG/invE de la Isla de Patogenicidad de Salmonella 1 (SPI-1) y de los genes ssaJ/ssaK en la SPI-2, se evaluaron los modelos asa intestinal ligada de ratón asociado a la observación de los tejidos por microscopía electrónica de transmisión (MET) y la producción de salmonelosis sistémica en ratón. Para ello, se utilizaron seis cepas de Salmonella: S. Typhimurium SL-1344 (cepa control) y sus derivadas mutantes: ∆invEG S. Typhimurium SL-1344 (mutante en SPI-1) y ∆ssaJK S. Typhimurium SL-1344 (mutante en SPI-2), S. Typhimurium (cepa clínica) y sus derivadas mutantes: ∆invEG S. Typhimurium y ∆ssaJK S. Typhimurium. Los resultados de MET permitieron verificar las alteraciones morfológicas del epitelio intestinal en el ratón infectado con cepas de Salmonella cuyos genes de patogenicidad estaban intactos. Fue comprobada la pérdida del poder invasivo solo en las cepas mutadas en la SPI-1. A través del modelo de salmonelosis sistémica en ratón se pudo comprobar la pérdida de la capacidad de diseminación en ambas mutantes. En conclusión los modelos permitieron verificar la importancia que tienen los genes invG/invE de la SPI-1 y ssaJ/ssaK de la SPI-2 en la patogenia de la salmonelosis, utilizando como modelo experimental de infección ratones BALB/c. Se sugieren estos modelos in vivo para evaluar mutantes de genes implicados en la patogenia de Salmonella, ya que representan una herramienta importante para la comprensión de la interacción Salmonella-hospedero(AU)

With the aim of evaluate the pathogenesis in Salmonella Typhimurium strains with mutations in genes invG/invE of Salmonella Pathogenicity Island 1 (SPI-1) and genes ssaJ/ssaK in the SPI-2 models were evaluated ligated intestinal loop associated mouse tissues by observation by transmission electron microscopy (TEM) and the production of mouse systemic salmonellosis. For this, we used six Salmonella strains: S. Typhimurium SL-1344 (control strain) and its derived mutants: ΔinvEG S. Typhimurium SL-1344 (mutant in SPI-1) and ΔssaJK S. Typhimurium SL-1344 (mutant in SPI-2), S. Typhimurium (clinical isolate) and its derived mutants: ΔinvEG S.Typhimurium and ΔssaJK S. Typhimurium. TEM results allowed us to verify the morphological alterations of the intestinal epithelium in mice infected with Salmonella strains whose pathogenicity genes were intact. It was proven invasive power loss only in strains mutated in the SPI-1. Through systemic salmonellosis model mouse we noted the loss of the ability to spread in both mutants. In conclusion, the models allowed us to verify the importance of the invG/invE genes of SPI-1 and ssaJ/ssaK of SPI-2 in the pathogenesis of salmonellosis, using BALB/c mice as an experimental model of infection. These in vivo models are suggested to evaluate mutants of genes involved in the pathogenesis of Salmonella, since they represent an important tool for the understanding of the Salmonella-host interaction(AU)

Phenotypic characterization of swine peripheral blood monocyte-derived macrophages and ex vivo infection with Salmonella enterica serovar Typhimurium.

Macrophages are critical mediators of the inflammatory process, playing a relevant role in the pathogenesis of Salmonella Typhimurium. The protocols for isolation, culture, and differentiation of monocytes into macrophages and their interaction with Salmonella are well established in humans and murine models, but little information is available in swine. The aims of this study were to establish an efficient protocol for macrophage culture and to evaluate the interaction of the invA mutant strain and the wild type (WT) Salmonella Typhimurium with porcine macrophages. Peripheral blood monocyte-derived macrophages from pigs were obtained, separated by density-gradient centrifugation, and cultured in Teflon vials for 10 days. After the differentiation period, cultures consisted of 92.4% CD14+ cells. In addition, these cells showed phagocytic ability, demonstrated by the presence of the same amount of WT and invA mutant Salmonella Typhimurium 1 h after interaction with macrophages. The early cytotoxic effect was Salmonella pathogenicity island (SPI)-[1]dependent, in which log-phase WT strains were more efficient (p < 0.01) than the invA mutant strain at inducing the death of macrophages.

Virulence Factors in Salmonella Typhimurium: The Sagacity of a Bacterium.

Currently, Salmonella enterica Typhimurium (ST) is responsible for most cases of food poisoning in several countries. It is characterized as a non-specific zoonotic bacterium that can infect both humans and animals and although most of the infections caused by this microorganism cause only a self-limiting gastroenteritis, some ST strains have been shown to be invasive, crossing the intestinal wall and reaching the systemic circulation. This unusual pathogenicity ability is closely related to ST virulence factors. This review aims to portray the main virulence factors in Salmonella Typhimurium, in order to better understand the strategies that this pathogen uses to reach the systemic circulation and increase its infectivity in humans and animals. Thus, the most studied Salmonella pathogenicity islands in Salmonella Typhimurium were detailed as to the functions of their encoded virulence factors. In addition, available knowledge on virulence plasmid was also compiled, as well as the chromosome regions involved in the virulence of this bacterium.