Introduction: Salmonella and Shigella are gram-negative bacilli that are resistant to most antibiotics and play an important role in an etiology of diarrhea disease. The aim of the study was to determine the prevalenc...Introduction: Salmonella and Shigella are gram-negative bacilli that are resistant to most antibiotics and play an important role in an etiology of diarrhea disease. The aim of the study was to determine the prevalence of Salmonella spp and Shigella spp isolated from stool and blood samples in the city of N’Djamena. Materials and Method: This was a prospective study conducted in the four district hospitals of N’Djamena from 14 July 2022 to 31 December 2022. A questionnaire form was drawn up to collect the information sent to the study patients. The samples were analyzed at the CHU de la Mère et de l’Enfant, Labo-Redes laboratory according to their protocols and the standard of the antibiogram committee of the French microbiology society. Results: Of the 803 biological samples analyzed, 39 were positive for Salmonella spp and Shigella spp, including 15 for Salmonella and 24 for Shigella, giving an overall prevalence rate of 4.85%. Borehole water, uncooked food and lack of access to a latrine constitute a risk of being infected by Salmonella spp and Shigella spp species. Of the 8 antibiotics tested, Salmonella spp and Shigella spp strains showed good sensitivity to nalidixic acid (100% for Salmonella and 90 for Shigella) and to ciprofloxacin (90.9% for Salmonella and 75% for Shigella). Resistance to ampicillin was found in 81.81% of Salmonella species and 78.57% of Shigella species, as was resistance to chloramphenicol (81.81% of Salmonella species and 67.85% of Shigella species). Similarly, cleanliness of the service and equipment is an essential factor in preventing Salmonella and Shigella infections.展开更多
Objective To investigate the characteristic of integrons and the relationship between integrons and antimicrobial resistance in Shigella spp. Methods Ninety Shigella strains (83 S. flexneri and 7 S. sonnei) were iso...Objective To investigate the characteristic of integrons and the relationship between integrons and antimicrobial resistance in Shigella spp. Methods Ninety Shigella strains (83 S. flexneri and 7 S. sonnei) were isolated from the stools of patients in China. Susceptibility to 8 antimicrobials was tested for all isolated strains. PCR, RFLP and sequencing analysis of integrons were applied to all of them. Results High prevalence of multi‐drug resistance (95.6%) was identified. Of the isolates 79 (87.8%) carried integrase genes of class 1 integron (3.3%), class 2 integron (10.0%) or both (74.4%). No intI3 was detected in the tested isolates. The prevalence of intI2 was significantly higher in isolates with multi‐drug resistance to at least 3 antibiotics than that in isolates with resistance to 2 and less antibiotics (P0.05). Gene cassettes dfrA17‐aadA5, dfrA12‐orfF‐aadA2 of class 1 integron and dfrA1‐sat1‐aadA1 of class 2 integron were identified. Conclusion The class 2 integron may play a role in the emergence of multi‐drug resistance in Shigella spp.展开更多
AIM: To profile the immunogenic proteins of Shigella flexneri (S. flexneri) expressed during human infection using a proteomic approach. METHODS: Soluble and membrane protein extractions of S. flexneri 2457T were sepa...AIM: To profile the immunogenic proteins of Shigella flexneri (S. flexneri) expressed during human infection using a proteomic approach. METHODS: Soluble and membrane protein extractions of S. flexneri 2457T were separated by two-dimensional gel electrophoresis (2-DE). Proteins were transferred to PVDF membrane and immunoblotted with sera from shigellosis patients. Reactive protein spots were matched to Coomassie stained gels run in parallel, cut out and trypsin digested. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) was used to determine the peptide mass fingerprints, which were searched in the MASCOT database to identify the protein. RESULTS: A total of 8 immunoreactive proteins were successfully identified from the Coomassie stained gels in three repeats. Six of these proteins have not previously been reported as immunogenic in S. flexneri. These proteins could be potential candidates for vaccine or attenuation studies. CONCLUSION: Soluble and membrane proteins of S. flexneri 2457T have been screened by 2-DE and immunoblotting with sera from shigellosis patients. Eight proteins are identified as immunogenic.展开更多
AIM: To screen the immunogenic membrane proteins of Shigella Aexneri 2a 2457T. METHODS: The routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and Western blotting were combined to screen immunogeni...AIM: To screen the immunogenic membrane proteins of Shigella Aexneri 2a 2457T. METHODS: The routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and Western blotting were combined to screen immunogenic proteins of S. Aexneri 2a 2457T. Serum was gained from rabbits immunized with the same bacteria. Immunogenic spots were cut out from the polyacrylamide gel and digested by trypsin in-gel. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) was performed to determine the molecular weight of peptides. Electrospray ionization (ESI-MS/MS) was performed to determine the sequences of the interesting peptides. RESULTS: A total of 20 spots were successfully identified from Coomassie brilliant blue stained gels representing 13 protein entries, 5 known antigens and 8 novel antigens. A hypothetical protein (YaeT) was detected, which might be a candidate target of vaccine. CONCLUSION: Membrane proteins of S. flexneri 2a 2457T were successfully observed by 2-DE. Several known and novel antigens were identified by mass spectrum.展开更多
Shigellosis causes diarrheal disease in humans in both developed and developing countries, and multi-drug resistance in Shigella is an emerging problem. Understanding changing resistance patterns is important in deter...Shigellosis causes diarrheal disease in humans in both developed and developing countries, and multi-drug resistance in Shigella is an emerging problem. Understanding changing resistance patterns is important in determining appropriate antibiotic treatments. This meta-analysis systematically evaluated aminoglycoside resistance in Shigella. A systematic review was constructed based on MEDLINE and EMBASE databases. Randomeffect models or fixed-effect models were used based on P value considering the possibility of heterogeneity between studies for meta-analysis. Data manipulation and statistical analyses were performed using software STATA 11.0. By means of meta-analysis, we found a lower resistance to three kinds of aminoglycosides in the Europe-America areas during the 12 year study period than that of the Asia-Africa areas. Kanamycin resistance was observed to be the most common drug resistance among Shigella isolates with a prevalence of 6.88% (95%CI: 6.36%-7.43%). Comparison of data from Europe-America and Asia-Africa areas revealed that Shigella flexneri resistance was greater than the resistance calculated for Shigella sonnei. Importantly, Shigella sonnei has played a significant role in aminoglycoside-resistance in recent years. Similarly, data showed that resistance to these drugs in children was higher than the corresponding data of adults. In conclusion, aminoglycoside-resistant Shigella is not an unusual phenomenon worldwide. Distribution in Shigella resistance differs sharply based on geographic areas, periods of time and subtypes. The results from the present study highlight the need for con- tinuous surveillance of resistance and control of antibiotic usage.展开更多
AIM: To construct and evaluate a polyvalent recombinant vaccine strain Shigella flexneri2a T32 against enterotoxigenic E.coli/(ETEC). METHODS: By using a host-plasmid balanced lethal system based on asd gene, a polyva...AIM: To construct and evaluate a polyvalent recombinant vaccine strain Shigella flexneri2a T32 against enterotoxigenic E.coli/(ETEC). METHODS: By using a host-plasmid balanced lethal system based on asd gene, a polyvalent recombinant strain was constructed to highly express CS3 and regularly express fusion enterotoxin of LIB subunit and mutant ST (LTB/STm) in a vaccine strain Shigella flexneri 2a T32 with specific deletion of asd gene. Fimbria CS3 was observed by immunofluorescence and electron microscopy assay. The security of LTB/STm was examined by ileal loop assay and suckling mouse assay. To evaluate this new candidate vaccine, it was compared with a previous vaccine strain in plasmid and protein level, growth assay and immunogenicity in Balb/c mice. RESULTS: The newly constructed vaccine expressed CS3 and grew better than the previously constructed vaccine except for the lower expression of LTB/STm. Serum IgG and mucosal IgA against CS3, LTB, ST, and host lipopolysaccharide (LPS) were produced after immunization of Balb/c mice by oral route with the new strain. The titers were not significantly different from the Balb/c mice with the previous strain. CONCLUSION: This novel candidate diarrheal vaccine can effectively induce serum and mucosal antibody responses against ETEC and Shigella.展开更多
AIM To investigate the specific pathogenesis ofO-polysaccharide (O--PS) which is on the outermembrane of lipopolysaccharides (LPS) fromShigella fi^eri.METHODS The O--PS was isolated and purifiedfrom Shigella nexneri 5...AIM To investigate the specific pathogenesis ofO-polysaccharide (O--PS) which is on the outermembrane of lipopolysaccharides (LPS) fromShigella fi^eri.METHODS The O--PS was isolated and purifiedfrom Shigella nexneri 5 MgoT by enzymatichydrolysis and gel chromatography. Effects ofO--PS were observed by in vitro experiment,(HeLa cell Culture ), and in vivo experiment(rabbit lieal loop assay).RESULTS ID vitro and in vivo e-cP6riments withthe purified O--PS from Shigells flexnefi revealedthat the O--PS alone was toxic to Hela cells andcaused mucosal inflammation and hemorrhagicexudation in lieal loop of rabbit.DISCUSSION O--PS might b6 one of the factorscausing diarrhea and its mechanism wasdifferent from endotoxin reaction of LPS. Themolecular mechanism of O-PS need furtherstudies.展开更多
Objective:To investigate the antimicrobial resistance patterns and prevalence of integrons in Shigella species isolated from children with diarrhea in southwest Iran.Methods:In this study,1530 stool samples were colle...Objective:To investigate the antimicrobial resistance patterns and prevalence of integrons in Shigella species isolated from children with diarrhea in southwest Iran.Methods:In this study,1530 stool samples were collected from children under 15 years with diarrhea referred to teaching hospitals in Ahvaz and Abadan,southwest Iran.Shigella spp.were identified by standard biochemical tests and PCR.The antibiotic resistance pattern of all Shigella isolates was determined by the disk diffusion method and minimum inhibitory concentration(MIC)by E-test.Results:Of 1530 stool samples,91(5.9%,91/1530)were positive for Shigella spp.the most common Shigella isolates were Shigella flexneri 47(51.6%,47/1530).Antibiotic susceptibility tests showed that the highest antibiotic resistance was related to trimethoprimsulfamethoxazole(87.9%,80/91)and ampicillin(86.8%,79/91).Multiplex PCR results revealed that 56%and 86.9%of Shigella isolates carried integron classⅠand integron classⅡgenes,respectively.None of the isolates included the integron classⅢgene.Conclusions:The high prevalence of multi-drug resistance in Shigella isolates in our area increases the concerns about dissemination of the antibiotic-resistant isolates in this bacterium.展开更多
[Objective] The research aimed to establish a rapid detection method for Shigella. [Method] Combining immunomagnetic separation technology with ATP bioluminescence technology, a new kind of fast and accurate ATP biolu...[Objective] The research aimed to establish a rapid detection method for Shigella. [Method] Combining immunomagnetic separation technology with ATP bioluminescence technology, a new kind of fast and accurate ATP bioluminescence magnetic enzyme immunoassay technique for Shigella was established. [Result] Using ATP bioluminescence magnetic enzyme immunoassay technique to detect standard solution for Shigella (ATCC 25931 ), result showed that correlation coefficient between relative light intensity detected by instrument and bacteria concentration detec- ted by culture counting method was 0.981 1. Moreover, relation curve between relative light intensity and Shigella concentration was drawn. [ Conclusion] The method had a high detection speed and accuracy, and could be used for the rapid detection of pathogen in food and environment.展开更多
Objective: We aimed to evaluate the combined antibacterial effects of allicin in combination with levofloxacin and ceftriaxone on Shigella isolated from the intestinal tract in vitro. Materials and Methods: Using a ch...Objective: We aimed to evaluate the combined antibacterial effects of allicin in combination with levofloxacin and ceftriaxone on Shigella isolated from the intestinal tract in vitro. Materials and Methods: Using a checkerboard design, broth microdilution assay was used to test the effects of the compounds on the organism. We also determined the MIC of the two groups of antibacterial drugs against 30 strains of Shigella and calculated the fractional inhibitory concentration(FIC) index, to judge the combination effect. Result: After the combined application of allicin and ceftriaxone the MIC decreased significantly. Distribution of the FIC index was as follows: FIC ≤0.5, accounting for 10%; 0.5< FIC ≤1.0, accounting for 60%; 1 < FIC ≤2, accounting for 30%; FIC >2, percentage is zero. After combined application of allicin and levofloxacin, distribution of FIC index was as follows: FIC≤0.5, ratio is zero; 0.5< FIC ≤1, accounting for 56.7%; 1 < FIC ≤2, accounting for 43.3%; FIC >2, ratio is zero. Conclusion: After the combined use of ceftriaxone, levofloxacin, and allicin, most of the tests showed synergistic effects and additive effects on Shigella, while some of them showed no correlation and no antagonistic effect.展开更多
Background: Shigella is one of the most serious pathogens associated with bloody diarrhea in children. The empiric antibiotic therapy of enteric illness with blood streaked stool leads to emergence of multi drug resis...Background: Shigella is one of the most serious pathogens associated with bloody diarrhea in children. The empiric antibiotic therapy of enteric illness with blood streaked stool leads to emergence of multi drug resistant (MDR) Shigella. The condition gets exacerbated by presence of integrons that facilitate the horizontal spread. Virulence genes associated with MDR Shigella modulate the patient outcome, particularly in children. Objectives: The present study was aiming at isolation of MDR Shigella from children with diarrheal sickness and characterization of those isolates as regarding presence of class 1 integrase and other virulence genes. Methods: Four hundred and ninety patients under the age of five suffering from diarrheal illness were examined for presence of Shigella in their stool specimens. MDR Shigella was determined using the antibiotic susceptibility testing by disc diffusion method;those isolates were tested for presence of class 1 integrase by PCR. Multiplex PCR assay was used to determine the presence of virulence genes, virA, ial, sen, set1A, set1B, sat, ipaBCD, ipaH and stx in the MDR Shigella isolates. Results: The isolation rate of Shigella from pediatric patients was 5.3%. Most of the isolated Shigella (57.7%) were from infants between 12 and 23 month. 73.1% of the identified Shigella were MDR. intI1 gene was present in 78.9% of MDR isolates. Muliplex PCR revealed that ipaH and ipaBCD, virA, sat, ial, set1A and set1B, sen were detected in 94.7%, 78.9%, 73.7%, 68.4%, 42.1%, 36.8% of the MDR Shigella isolates respectively. Conclusion: The MDR isolates represented a considerable percentage of Shigella detected in pediatric patients. Presence of intI1 gene in most of MDR Shigella reflects the higher possibility of resistant strains spread. Existence of a variety of virulence genes in those isolates is an important indicator of serious disease outcome.展开更多
An isothermal amplification assay for the detection of Shigella based colloidal gold strip detection is developed. The primers designed corresponded to the ipaH gene of Shigella .The sensitivity of the colloidal gold ...An isothermal amplification assay for the detection of Shigella based colloidal gold strip detection is developed. The primers designed corresponded to the ipaH gene of Shigella .The sensitivity of the colloidal gold strip-NASBA method in this study was 6.3 x 101cfu/ml. The specificity of the detection system was detected and the result showed that the NASBA method could distinguish Shigella from other germs.展开更多
Objective:To investigate the phytochemical components of Abrus precatorius(A.precatorius) and the in-vitro susceptibility of Salmonella typhi and Shigella dysenteriae to the aqueous extracts of A.precatorius leaf,seed...Objective:To investigate the phytochemical components of Abrus precatorius(A.precatorius) and the in-vitro susceptibility of Salmonella typhi and Shigella dysenteriae to the aqueous extracts of A.precatorius leaf,seed and root.Methods:The leaf,seed and root of A.precatorius were collected and homogenized separately after drying at 40℃ for seven days in hot-air oven.The aqueous extracts of each of the parts were prepared and subjected to phytochemical screening.Dilutions of 400,300,200,100 mg/mL,of each of the extracts were used for broth dilution in minimum inhibitory concentration(MIC) determination against clinical isolates of Salmonella typhi and Shigella dysenteriae,while 50,40,30,20,and 10 mg/mL dilutions were used for the agar diffusion test and 100 mg/mL and 10 mg/mL of gentamycin were used as controls for broth dilution in MIC determination and agar diffusion test,respectively.Results:Qualitative study reveals that tannin,saponins,alkaloids,flavonoids,terpenoids,steroids and phenols were present in all of the plant parts.The leaf has the highest quantities of tannin and phenol.The root generally showed the lowest quantity of all the compounds.The pathogens were susceptible to aqueous extracts of the leaf,stem and root of A.precatorius at 50 mg/mL.At concentrations of 40,30 and 20 mg/mL,all the aqueous extracts of A.precatorius showed variation in MIC,but produced no minimum bactericide effect upon subculture.There were variations in diameter of zone of inhibition against the organisms at lower concentrations.Conclusions:These findings suggest that A.precatorius is a valuable source of phytochemicals with promising antibacterial activity.Considering this bioactivity,A.precatorius could be probed further for toxicity,and to obtain some novel antibacterial molecules.展开更多
Objective:To evaluate the phytochemical and anti-bacterial efficacy of the seed kernel extract of Mangifera indica(M.indica) against the enteropathogen,Shigella dysenteriae(S.dysenteriae), isolated from the diarrhoeal...Objective:To evaluate the phytochemical and anti-bacterial efficacy of the seed kernel extract of Mangifera indica(M.indica) against the enteropathogen,Shigella dysenteriae(S.dysenteriae), isolated from the diarrhoeal stool specimens.Methods:The preliminary phytochemical screening was performed by the standard methods as described by Harborne.Cold extraction method was employed to extract the bioactive compounds from mango seed kernel.Disc diffusion method was adopted to screen antibacterial activity.Minimum inhibitory concentration(MIC) was evaluated by agar dilution method.The crude extracts were partially purified by thin layer chromatography(TLC) and the fractions were analyzed by high performance thin layer chromatography(HFTLC) to identify the bioactive compounds.Results:Phytochemical scrutiny of M.indica indicated the presence of phytochemical constituents such as alkaloids,gums, flavanoids,phenols,saponins,steroids,tannins and xanthoproteins.Antibacterial activity was observed in two crude extracts and various fractions viz.hexane,benzene,chlor of orm,methanol and water.MIC of methanol fraction was found to be(95±11.8)μg/mL.MIC of other fractions ranged from 130-380μg/mL Conclusions:The present study confirmed that each crude extracts and fractions of M.indica have significant antimicrobial activity against the isolated pathogen 5. dyserUeriae.The antibacterial activity may be due to the phytochemical constituents of the mango seed kernel.The phytochemical tannin could be the reason for its antibacterial activity.展开更多
BACKGROUND Shigella flexneri(S.flexneri)is a major pathogen causing acute intestinal infection,but the systematic oxidative damage incurred during the course of infection has not been investigated.AIM To investigate t...BACKGROUND Shigella flexneri(S.flexneri)is a major pathogen causing acute intestinal infection,but the systematic oxidative damage incurred during the course of infection has not been investigated.AIM To investigate the incurred systemic RNA oxidative damage and the diagnostic value of RNA oxidative metabolites during S.flexneri-induced intestinal infection.METHODS In this study,a Sprague-Dawley rat model of acute intestinal infection was established by oral gavage with S.flexneri strains.The changes in white blood cells(WBCs)and cytokine levels in blood and the inflammatory response in the colon were investigated.We also detected the RNA and DNA oxidation in urine and tissues.RESULTS S.flexneri infection induced an increase in WBCs,C-reactive protein,interleukin(IL)-6,IL-10,IL-1β,IL-4,IL-17a,IL-10,and tumor necrosis factorα(TNF-α)in blood.Of note,a significant increase in urinary 8-oxo-7,8-dihydroguanosine(8-oxo-Gsn),an important marker of total RNA oxidation,was detected after intestinal infection(P=0.03).The urinary 8-oxo-Gsn level returned to the baseline level after recovery from infection.In addition,the results of a correlation analysis showed that urinary 8-oxo-Gsn was positively correlated with the WBC count and the cytokines IL-6,TNF-α,IL-10,IL-1β,and IL-17α.Further detection of the oxidation in different tissues showed that S.flexneri infection induced RNA oxidative damage in the colon,ileum,liver,spleen,and brain.CONCLUSION Acute infection induced by S.flexneri causes increased RNA oxidative damage in various tissues(liver,spleen,and brain)and an increase of 8-oxo-Gsn,a urinary metabolite.Urinary 8-oxo-Gsn may be useful as a biomarker for evaluating the severity and prognosis of infection.展开更多
AIM: TO investigate the effect of interaction between enteric epithelial cells and lymphocytes of Peyer's patch on the release of nitric oxide (NO) and IL-6 in response to Shigella lipopolysaccharide (LPS). METH...AIM: TO investigate the effect of interaction between enteric epithelial cells and lymphocytes of Peyer's patch on the release of nitric oxide (NO) and IL-6 in response to Shigella lipopolysaccharide (LPS). METHODS: Human colonic epithelial cells (Caco-2) were mixed cocultured with lymphocytes of Peyer's patch from wild-type (C57 mice) and inducible NO synthase knockout mice, and challenged with Shigella F2a-12 LPS. Release of NO and raiL-6 was measured by Griess colorimetric assay and enzyme-linked immunosorbent assay (ELISA), respectively. RESULTS: In the absence of LPS challenge, NO was detected in the culture medium of Caco-2 epithelial cells but not in lymphocytes of Peyer's patch, and the NO release was further up-regulated in both cocultures with lymphocytes from either the wild-type or iNOS knockout mice, with a significantly higher level observed in the coculture with iNOS knockout lymphocytes. After Shigella F2a-12 LPS challenge for 24-h, NO production was significantly increased in both Caco-2 alone and the coculture with lymphocytes of Peyer's patch from the wild-type mice but not from iNOS knockout mice. LPS was found to stimulate the release of mIL-6 from lymphocytes, which was suppressed by coculture with Caco-2 epithelial cells. The LPS-induced mIL-6 production in lymphocytes from iNOS knockout mice was significantly greater than that from the wild-type mice. CONCLUSION: Lymphocytes of Peyer's patch maintain a constitutive basal level of NO production from the enteric epithelial cell Caco-2. LPS-induced mIL-6 release from lymphocytes of Peyer's patch is suppressed by the cocultured epithelial cells. While no changes are detectable in NO production in lymphocytes from both wild-type and iNOS knockout mice before and after LPS challenge, NO from lymphocytes appears to play an inhibitory role in epithelial NO release and their own mIL-6 release in response to LPS.展开更多
Aim: Shigella flexneri (S. flexneri) is a gram-negative enterobacterium responsible for severe intestinal end systemic infection in humans. The bacteria can reach the liver due to degeneration of the colonic epitheliu...Aim: Shigella flexneri (S. flexneri) is a gram-negative enterobacterium responsible for severe intestinal end systemic infection in humans. The bacteria can reach the liver due to degeneration of the colonic epithelium. Hypoxia is present in many human diseases and can induce the expression of the transcription factor HIF-1alpha that may have a cell protective role. The influence of hypoxia and HIF-1alpha on bacterial infection, studied in this work, is unclear. Hypoxia inducible factor-1alpha (HIF-1alpha) is a transcription factor that acts as a master regulator of gene expression induced by hypoxia. Methods: We compared the ability of S. flexneri to invade rat hepatocytes in primary culture both in normoxic and hypoxic conditions. We evaluated TNF-alpha released by hepatocytes, apoptosis rate and HIF-1alpha expression by confocal microscopy as well as real time PCR technique. Results: We showed that S. flexneri invaded less hepatocytes previously submitted to 24 h hypoxia (6.5% O2) than those cultivated in normoxia (21% O2). S. flexneri also induced HIF-1α expression in hepatocytes, TNF-α secretion and apoptosis. Conclusion: a) Hypoxia alone was not a stimulus to TNF-α secretion, but induced cell apoptosis and HIF-1α expression;b) S. flexneri was able to invade rat hepatocytes and hypoxia apparently influenced significantly bacterial cell invasiveness;c) HIF-1α was expressed in hypoxic conditions, and it was also stimulated by S. flexneri.展开更多
118 clinical strains of Shigella were serotyped, in which 116 strains were tested to be S. flexneri. The susceptibilities of the S .flexneri strains to quinolones were measured by the disk-diffusion method. It was fou...118 clinical strains of Shigella were serotyped, in which 116 strains were tested to be S. flexneri. The susceptibilities of the S .flexneri strains to quinolones were measured by the disk-diffusion method. It was found that most S .flexneri strains were susceptible to norfloxacin and ciprofloxacin, but resistant to nalidixic acid. To study the correlation between gyrA mutations and quinolones resistance, a fragment within the gyrA gene was amplified by PCR. The SSCP (Single-Strand Conformation Polymorphism) analysis was applied to detect mutations in PCR products of different strains. The mutations were then confirmed by DNA sequencing. Altogether, two types of mutation were revealed, in which one type was single mutation ( C42-T), and the other was double mutations ( C42-T and A54- G). By statistical analysis, C42-T (encoding Ser83-keu substitution) was shown to have correlation with nalidixic-acid resistance in the clinical strains of Shigella, while A54-G (encoding Asp87-Gly substitution) was shown to have correlations with both norfloxacin resistance and ciprofloxacin resistance.展开更多
According to the World Health Organization, foodborne diseases are a major public health problem, particularly in developing countries including the Republic of Congo. They are responsible for several episodes of diar...According to the World Health Organization, foodborne diseases are a major public health problem, particularly in developing countries including the Republic of Congo. They are responsible for several episodes of diarrhea, especially in children under five years old. There is no reliable epidemiological data on the pathogenicity of the Shigella spp. strains circulating in the whole Republic of Congo drafting this paper. The purpose of this study was to examine the Shigella spp strain pathogenicity close to an environment contaminated with faeces in the city of Brazzaville. As a result, 54 isolates have been associated with Shigella spp. The gastric acid resistance test performed on Shigella Environmental Strain (SES) and Shigella Clinical Strains (SCS) resulted in 38.8% (21/54) and 100% acid resistant, respectively. Shigella spp. Strains (SES and SCS) were ranged in a survival percentage from 11% to 93%. By monitoring Biosurfactant-Like Molecule (BLM) production, we showed that the BLM production of SES and SCS was highly dependent on bacterial culture density involving the Quorum Sensing (QS). S. flexneri, S. boydii, and S. sonnei and as well as SES and SCS were able to invade and contaminate eggs by colonizing egg yolk. The counting bacteria were ranging from two to 5 × 10<sup>7</sup> CFU/mL after contamination. Concomitantly, BLM was secreted during the post contamination of poultry eggs with 100% EI24. Further by trying to show the pathogenicity by the hemolysis test, we have shown that SES and SCS were able to show significant areas of lysis on blood agar. Finally, this work has proposed an additional model of cell invasion including biosurfactants during the pre- and post-invasion phases.展开更多
文摘Introduction: Salmonella and Shigella are gram-negative bacilli that are resistant to most antibiotics and play an important role in an etiology of diarrhea disease. The aim of the study was to determine the prevalence of Salmonella spp and Shigella spp isolated from stool and blood samples in the city of N’Djamena. Materials and Method: This was a prospective study conducted in the four district hospitals of N’Djamena from 14 July 2022 to 31 December 2022. A questionnaire form was drawn up to collect the information sent to the study patients. The samples were analyzed at the CHU de la Mère et de l’Enfant, Labo-Redes laboratory according to their protocols and the standard of the antibiogram committee of the French microbiology society. Results: Of the 803 biological samples analyzed, 39 were positive for Salmonella spp and Shigella spp, including 15 for Salmonella and 24 for Shigella, giving an overall prevalence rate of 4.85%. Borehole water, uncooked food and lack of access to a latrine constitute a risk of being infected by Salmonella spp and Shigella spp species. Of the 8 antibiotics tested, Salmonella spp and Shigella spp strains showed good sensitivity to nalidixic acid (100% for Salmonella and 90 for Shigella) and to ciprofloxacin (90.9% for Salmonella and 75% for Shigella). Resistance to ampicillin was found in 81.81% of Salmonella species and 78.57% of Shigella species, as was resistance to chloramphenicol (81.81% of Salmonella species and 67.85% of Shigella species). Similarly, cleanliness of the service and equipment is an essential factor in preventing Salmonella and Shigella infections.
基金supported by the Research Fund from Ministry of Health of the People’s Republic of China, WKJ2007‐2‐024
文摘Objective To investigate the characteristic of integrons and the relationship between integrons and antimicrobial resistance in Shigella spp. Methods Ninety Shigella strains (83 S. flexneri and 7 S. sonnei) were isolated from the stools of patients in China. Susceptibility to 8 antimicrobials was tested for all isolated strains. PCR, RFLP and sequencing analysis of integrons were applied to all of them. Results High prevalence of multi‐drug resistance (95.6%) was identified. Of the isolates 79 (87.8%) carried integrase genes of class 1 integron (3.3%), class 2 integron (10.0%) or both (74.4%). No intI3 was detected in the tested isolates. The prevalence of intI2 was significantly higher in isolates with multi‐drug resistance to at least 3 antibiotics than that in isolates with resistance to 2 and less antibiotics (P0.05). Gene cassettes dfrA17‐aadA5, dfrA12‐orfF‐aadA2 of class 1 integron and dfrA1‐sat1‐aadA1 of class 2 integron were identified. Conclusion The class 2 integron may play a role in the emergence of multi‐drug resistance in Shigella spp.
文摘AIM: To profile the immunogenic proteins of Shigella flexneri (S. flexneri) expressed during human infection using a proteomic approach. METHODS: Soluble and membrane protein extractions of S. flexneri 2457T were separated by two-dimensional gel electrophoresis (2-DE). Proteins were transferred to PVDF membrane and immunoblotted with sera from shigellosis patients. Reactive protein spots were matched to Coomassie stained gels run in parallel, cut out and trypsin digested. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) was used to determine the peptide mass fingerprints, which were searched in the MASCOT database to identify the protein. RESULTS: A total of 8 immunoreactive proteins were successfully identified from the Coomassie stained gels in three repeats. Six of these proteins have not previously been reported as immunogenic in S. flexneri. These proteins could be potential candidates for vaccine or attenuation studies. CONCLUSION: Soluble and membrane proteins of S. flexneri 2457T have been screened by 2-DE and immunoblotting with sera from shigellosis patients. Eight proteins are identified as immunogenic.
基金Supported by the Capital "248" Key Innovation Project, No. H010210360119, State Basic Research Development Program of China No. 973 Program, G1999054103 and 2005CB22904 and National Natural Science Foundation of China No. 30470101
文摘AIM: To screen the immunogenic membrane proteins of Shigella Aexneri 2a 2457T. METHODS: The routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and Western blotting were combined to screen immunogenic proteins of S. Aexneri 2a 2457T. Serum was gained from rabbits immunized with the same bacteria. Immunogenic spots were cut out from the polyacrylamide gel and digested by trypsin in-gel. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) was performed to determine the molecular weight of peptides. Electrospray ionization (ESI-MS/MS) was performed to determine the sequences of the interesting peptides. RESULTS: A total of 20 spots were successfully identified from Coomassie brilliant blue stained gels representing 13 protein entries, 5 known antigens and 8 novel antigens. A hypothetical protein (YaeT) was detected, which might be a candidate target of vaccine. CONCLUSION: Membrane proteins of S. flexneri 2a 2457T were successfully observed by 2-DE. Several known and novel antigens were identified by mass spectrum.
基金funded by National Natural Science Foundation of China (No. 81000754)a grant from the Key Laboratory for Laboratory Medicine of Jiangsu Province of China (No. XK201114)a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘Shigellosis causes diarrheal disease in humans in both developed and developing countries, and multi-drug resistance in Shigella is an emerging problem. Understanding changing resistance patterns is important in determining appropriate antibiotic treatments. This meta-analysis systematically evaluated aminoglycoside resistance in Shigella. A systematic review was constructed based on MEDLINE and EMBASE databases. Randomeffect models or fixed-effect models were used based on P value considering the possibility of heterogeneity between studies for meta-analysis. Data manipulation and statistical analyses were performed using software STATA 11.0. By means of meta-analysis, we found a lower resistance to three kinds of aminoglycosides in the Europe-America areas during the 12 year study period than that of the Asia-Africa areas. Kanamycin resistance was observed to be the most common drug resistance among Shigella isolates with a prevalence of 6.88% (95%CI: 6.36%-7.43%). Comparison of data from Europe-America and Asia-Africa areas revealed that Shigella flexneri resistance was greater than the resistance calculated for Shigella sonnei. Importantly, Shigella sonnei has played a significant role in aminoglycoside-resistance in recent years. Similarly, data showed that resistance to these drugs in children was higher than the corresponding data of adults. In conclusion, aminoglycoside-resistant Shigella is not an unusual phenomenon worldwide. Distribution in Shigella resistance differs sharply based on geographic areas, periods of time and subtypes. The results from the present study highlight the need for con- tinuous surveillance of resistance and control of antibiotic usage.
基金Supported by the National High Technology Research and Development Program of China (863 Program), No. 2001AA215211the Military Basic Research Foundation, No. 01Z026
文摘AIM: To construct and evaluate a polyvalent recombinant vaccine strain Shigella flexneri2a T32 against enterotoxigenic E.coli/(ETEC). METHODS: By using a host-plasmid balanced lethal system based on asd gene, a polyvalent recombinant strain was constructed to highly express CS3 and regularly express fusion enterotoxin of LIB subunit and mutant ST (LTB/STm) in a vaccine strain Shigella flexneri 2a T32 with specific deletion of asd gene. Fimbria CS3 was observed by immunofluorescence and electron microscopy assay. The security of LTB/STm was examined by ileal loop assay and suckling mouse assay. To evaluate this new candidate vaccine, it was compared with a previous vaccine strain in plasmid and protein level, growth assay and immunogenicity in Balb/c mice. RESULTS: The newly constructed vaccine expressed CS3 and grew better than the previously constructed vaccine except for the lower expression of LTB/STm. Serum IgG and mucosal IgA against CS3, LTB, ST, and host lipopolysaccharide (LPS) were produced after immunization of Balb/c mice by oral route with the new strain. The titers were not significantly different from the Balb/c mice with the previous strain. CONCLUSION: This novel candidate diarrheal vaccine can effectively induce serum and mucosal antibody responses against ETEC and Shigella.
基金Project supported by the National Natural Science Foundation of China,No.39370040.
文摘AIM To investigate the specific pathogenesis ofO-polysaccharide (O--PS) which is on the outermembrane of lipopolysaccharides (LPS) fromShigella fi^eri.METHODS The O--PS was isolated and purifiedfrom Shigella nexneri 5 MgoT by enzymatichydrolysis and gel chromatography. Effects ofO--PS were observed by in vitro experiment,(HeLa cell Culture ), and in vivo experiment(rabbit lieal loop assay).RESULTS ID vitro and in vivo e-cP6riments withthe purified O--PS from Shigells flexnefi revealedthat the O--PS alone was toxic to Hela cells andcaused mucosal inflammation and hemorrhagicexudation in lieal loop of rabbit.DISCUSSION O--PS might b6 one of the factorscausing diarrhea and its mechanism wasdifferent from endotoxin reaction of LPS. Themolecular mechanism of O-PS need furtherstudies.
基金supported by the Vice-Chancellor for Research grant(Grant No.U98-564)of Abadan University of Medical Science
文摘Objective:To investigate the antimicrobial resistance patterns and prevalence of integrons in Shigella species isolated from children with diarrhea in southwest Iran.Methods:In this study,1530 stool samples were collected from children under 15 years with diarrhea referred to teaching hospitals in Ahvaz and Abadan,southwest Iran.Shigella spp.were identified by standard biochemical tests and PCR.The antibiotic resistance pattern of all Shigella isolates was determined by the disk diffusion method and minimum inhibitory concentration(MIC)by E-test.Results:Of 1530 stool samples,91(5.9%,91/1530)were positive for Shigella spp.the most common Shigella isolates were Shigella flexneri 47(51.6%,47/1530).Antibiotic susceptibility tests showed that the highest antibiotic resistance was related to trimethoprimsulfamethoxazole(87.9%,80/91)and ampicillin(86.8%,79/91).Multiplex PCR results revealed that 56%and 86.9%of Shigella isolates carried integron classⅠand integron classⅡgenes,respectively.None of the isolates included the integron classⅢgene.Conclusions:The high prevalence of multi-drug resistance in Shigella isolates in our area increases the concerns about dissemination of the antibiotic-resistant isolates in this bacterium.
文摘[Objective] The research aimed to establish a rapid detection method for Shigella. [Method] Combining immunomagnetic separation technology with ATP bioluminescence technology, a new kind of fast and accurate ATP bioluminescence magnetic enzyme immunoassay technique for Shigella was established. [Result] Using ATP bioluminescence magnetic enzyme immunoassay technique to detect standard solution for Shigella (ATCC 25931 ), result showed that correlation coefficient between relative light intensity detected by instrument and bacteria concentration detec- ted by culture counting method was 0.981 1. Moreover, relation curve between relative light intensity and Shigella concentration was drawn. [ Conclusion] The method had a high detection speed and accuracy, and could be used for the rapid detection of pathogen in food and environment.
文摘Objective: We aimed to evaluate the combined antibacterial effects of allicin in combination with levofloxacin and ceftriaxone on Shigella isolated from the intestinal tract in vitro. Materials and Methods: Using a checkerboard design, broth microdilution assay was used to test the effects of the compounds on the organism. We also determined the MIC of the two groups of antibacterial drugs against 30 strains of Shigella and calculated the fractional inhibitory concentration(FIC) index, to judge the combination effect. Result: After the combined application of allicin and ceftriaxone the MIC decreased significantly. Distribution of the FIC index was as follows: FIC ≤0.5, accounting for 10%; 0.5< FIC ≤1.0, accounting for 60%; 1 < FIC ≤2, accounting for 30%; FIC >2, percentage is zero. After combined application of allicin and levofloxacin, distribution of FIC index was as follows: FIC≤0.5, ratio is zero; 0.5< FIC ≤1, accounting for 56.7%; 1 < FIC ≤2, accounting for 43.3%; FIC >2, ratio is zero. Conclusion: After the combined use of ceftriaxone, levofloxacin, and allicin, most of the tests showed synergistic effects and additive effects on Shigella, while some of them showed no correlation and no antagonistic effect.
文摘Background: Shigella is one of the most serious pathogens associated with bloody diarrhea in children. The empiric antibiotic therapy of enteric illness with blood streaked stool leads to emergence of multi drug resistant (MDR) Shigella. The condition gets exacerbated by presence of integrons that facilitate the horizontal spread. Virulence genes associated with MDR Shigella modulate the patient outcome, particularly in children. Objectives: The present study was aiming at isolation of MDR Shigella from children with diarrheal sickness and characterization of those isolates as regarding presence of class 1 integrase and other virulence genes. Methods: Four hundred and ninety patients under the age of five suffering from diarrheal illness were examined for presence of Shigella in their stool specimens. MDR Shigella was determined using the antibiotic susceptibility testing by disc diffusion method;those isolates were tested for presence of class 1 integrase by PCR. Multiplex PCR assay was used to determine the presence of virulence genes, virA, ial, sen, set1A, set1B, sat, ipaBCD, ipaH and stx in the MDR Shigella isolates. Results: The isolation rate of Shigella from pediatric patients was 5.3%. Most of the isolated Shigella (57.7%) were from infants between 12 and 23 month. 73.1% of the identified Shigella were MDR. intI1 gene was present in 78.9% of MDR isolates. Muliplex PCR revealed that ipaH and ipaBCD, virA, sat, ial, set1A and set1B, sen were detected in 94.7%, 78.9%, 73.7%, 68.4%, 42.1%, 36.8% of the MDR Shigella isolates respectively. Conclusion: The MDR isolates represented a considerable percentage of Shigella detected in pediatric patients. Presence of intI1 gene in most of MDR Shigella reflects the higher possibility of resistant strains spread. Existence of a variety of virulence genes in those isolates is an important indicator of serious disease outcome.
文摘An isothermal amplification assay for the detection of Shigella based colloidal gold strip detection is developed. The primers designed corresponded to the ipaH gene of Shigella .The sensitivity of the colloidal gold strip-NASBA method in this study was 6.3 x 101cfu/ml. The specificity of the detection system was detected and the result showed that the NASBA method could distinguish Shigella from other germs.
基金Supported by Institutional Based Research grant from the Tertiary Education Trust Fund,Nigeria(TETFUND/KWASU-2014-01)
文摘Objective:To investigate the phytochemical components of Abrus precatorius(A.precatorius) and the in-vitro susceptibility of Salmonella typhi and Shigella dysenteriae to the aqueous extracts of A.precatorius leaf,seed and root.Methods:The leaf,seed and root of A.precatorius were collected and homogenized separately after drying at 40℃ for seven days in hot-air oven.The aqueous extracts of each of the parts were prepared and subjected to phytochemical screening.Dilutions of 400,300,200,100 mg/mL,of each of the extracts were used for broth dilution in minimum inhibitory concentration(MIC) determination against clinical isolates of Salmonella typhi and Shigella dysenteriae,while 50,40,30,20,and 10 mg/mL dilutions were used for the agar diffusion test and 100 mg/mL and 10 mg/mL of gentamycin were used as controls for broth dilution in MIC determination and agar diffusion test,respectively.Results:Qualitative study reveals that tannin,saponins,alkaloids,flavonoids,terpenoids,steroids and phenols were present in all of the plant parts.The leaf has the highest quantities of tannin and phenol.The root generally showed the lowest quantity of all the compounds.The pathogens were susceptible to aqueous extracts of the leaf,stem and root of A.precatorius at 50 mg/mL.At concentrations of 40,30 and 20 mg/mL,all the aqueous extracts of A.precatorius showed variation in MIC,but produced no minimum bactericide effect upon subculture.There were variations in diameter of zone of inhibition against the organisms at lower concentrations.Conclusions:These findings suggest that A.precatorius is a valuable source of phytochemicals with promising antibacterial activity.Considering this bioactivity,A.precatorius could be probed further for toxicity,and to obtain some novel antibacterial molecules.
文摘Objective:To evaluate the phytochemical and anti-bacterial efficacy of the seed kernel extract of Mangifera indica(M.indica) against the enteropathogen,Shigella dysenteriae(S.dysenteriae), isolated from the diarrhoeal stool specimens.Methods:The preliminary phytochemical screening was performed by the standard methods as described by Harborne.Cold extraction method was employed to extract the bioactive compounds from mango seed kernel.Disc diffusion method was adopted to screen antibacterial activity.Minimum inhibitory concentration(MIC) was evaluated by agar dilution method.The crude extracts were partially purified by thin layer chromatography(TLC) and the fractions were analyzed by high performance thin layer chromatography(HFTLC) to identify the bioactive compounds.Results:Phytochemical scrutiny of M.indica indicated the presence of phytochemical constituents such as alkaloids,gums, flavanoids,phenols,saponins,steroids,tannins and xanthoproteins.Antibacterial activity was observed in two crude extracts and various fractions viz.hexane,benzene,chlor of orm,methanol and water.MIC of methanol fraction was found to be(95±11.8)μg/mL.MIC of other fractions ranged from 130-380μg/mL Conclusions:The present study confirmed that each crude extracts and fractions of M.indica have significant antimicrobial activity against the isolated pathogen 5. dyserUeriae.The antibacterial activity may be due to the phytochemical constituents of the mango seed kernel.The phytochemical tannin could be the reason for its antibacterial activity.
基金Supported by the National Key R&D Program of China,No.2018YFC2000300and CAMS Innovation Fund for Medical Sciences,No.2018-I2M-1-002.
文摘BACKGROUND Shigella flexneri(S.flexneri)is a major pathogen causing acute intestinal infection,but the systematic oxidative damage incurred during the course of infection has not been investigated.AIM To investigate the incurred systemic RNA oxidative damage and the diagnostic value of RNA oxidative metabolites during S.flexneri-induced intestinal infection.METHODS In this study,a Sprague-Dawley rat model of acute intestinal infection was established by oral gavage with S.flexneri strains.The changes in white blood cells(WBCs)and cytokine levels in blood and the inflammatory response in the colon were investigated.We also detected the RNA and DNA oxidation in urine and tissues.RESULTS S.flexneri infection induced an increase in WBCs,C-reactive protein,interleukin(IL)-6,IL-10,IL-1β,IL-4,IL-17a,IL-10,and tumor necrosis factorα(TNF-α)in blood.Of note,a significant increase in urinary 8-oxo-7,8-dihydroguanosine(8-oxo-Gsn),an important marker of total RNA oxidation,was detected after intestinal infection(P=0.03).The urinary 8-oxo-Gsn level returned to the baseline level after recovery from infection.In addition,the results of a correlation analysis showed that urinary 8-oxo-Gsn was positively correlated with the WBC count and the cytokines IL-6,TNF-α,IL-10,IL-1β,and IL-17α.Further detection of the oxidation in different tissues showed that S.flexneri infection induced RNA oxidative damage in the colon,ileum,liver,spleen,and brain.CONCLUSION Acute infection induced by S.flexneri causes increased RNA oxidative damage in various tissues(liver,spleen,and brain)and an increase of 8-oxo-Gsn,a urinary metabolite.Urinary 8-oxo-Gsn may be useful as a biomarker for evaluating the severity and prognosis of infection.
基金Supported by Strategic Program of Chinese University of Hong KongDistinguished Young Investigator Fund of the National Natural Science Foundation of China, No. 30029002
文摘AIM: TO investigate the effect of interaction between enteric epithelial cells and lymphocytes of Peyer's patch on the release of nitric oxide (NO) and IL-6 in response to Shigella lipopolysaccharide (LPS). METHODS: Human colonic epithelial cells (Caco-2) were mixed cocultured with lymphocytes of Peyer's patch from wild-type (C57 mice) and inducible NO synthase knockout mice, and challenged with Shigella F2a-12 LPS. Release of NO and raiL-6 was measured by Griess colorimetric assay and enzyme-linked immunosorbent assay (ELISA), respectively. RESULTS: In the absence of LPS challenge, NO was detected in the culture medium of Caco-2 epithelial cells but not in lymphocytes of Peyer's patch, and the NO release was further up-regulated in both cocultures with lymphocytes from either the wild-type or iNOS knockout mice, with a significantly higher level observed in the coculture with iNOS knockout lymphocytes. After Shigella F2a-12 LPS challenge for 24-h, NO production was significantly increased in both Caco-2 alone and the coculture with lymphocytes of Peyer's patch from the wild-type mice but not from iNOS knockout mice. LPS was found to stimulate the release of mIL-6 from lymphocytes, which was suppressed by coculture with Caco-2 epithelial cells. The LPS-induced mIL-6 production in lymphocytes from iNOS knockout mice was significantly greater than that from the wild-type mice. CONCLUSION: Lymphocytes of Peyer's patch maintain a constitutive basal level of NO production from the enteric epithelial cell Caco-2. LPS-induced mIL-6 release from lymphocytes of Peyer's patch is suppressed by the cocultured epithelial cells. While no changes are detectable in NO production in lymphocytes from both wild-type and iNOS knockout mice before and after LPS challenge, NO from lymphocytes appears to play an inhibitory role in epithelial NO release and their own mIL-6 release in response to LPS.
文摘Aim: Shigella flexneri (S. flexneri) is a gram-negative enterobacterium responsible for severe intestinal end systemic infection in humans. The bacteria can reach the liver due to degeneration of the colonic epithelium. Hypoxia is present in many human diseases and can induce the expression of the transcription factor HIF-1alpha that may have a cell protective role. The influence of hypoxia and HIF-1alpha on bacterial infection, studied in this work, is unclear. Hypoxia inducible factor-1alpha (HIF-1alpha) is a transcription factor that acts as a master regulator of gene expression induced by hypoxia. Methods: We compared the ability of S. flexneri to invade rat hepatocytes in primary culture both in normoxic and hypoxic conditions. We evaluated TNF-alpha released by hepatocytes, apoptosis rate and HIF-1alpha expression by confocal microscopy as well as real time PCR technique. Results: We showed that S. flexneri invaded less hepatocytes previously submitted to 24 h hypoxia (6.5% O2) than those cultivated in normoxia (21% O2). S. flexneri also induced HIF-1α expression in hepatocytes, TNF-α secretion and apoptosis. Conclusion: a) Hypoxia alone was not a stimulus to TNF-α secretion, but induced cell apoptosis and HIF-1α expression;b) S. flexneri was able to invade rat hepatocytes and hypoxia apparently influenced significantly bacterial cell invasiveness;c) HIF-1α was expressed in hypoxic conditions, and it was also stimulated by S. flexneri.
文摘118 clinical strains of Shigella were serotyped, in which 116 strains were tested to be S. flexneri. The susceptibilities of the S .flexneri strains to quinolones were measured by the disk-diffusion method. It was found that most S .flexneri strains were susceptible to norfloxacin and ciprofloxacin, but resistant to nalidixic acid. To study the correlation between gyrA mutations and quinolones resistance, a fragment within the gyrA gene was amplified by PCR. The SSCP (Single-Strand Conformation Polymorphism) analysis was applied to detect mutations in PCR products of different strains. The mutations were then confirmed by DNA sequencing. Altogether, two types of mutation were revealed, in which one type was single mutation ( C42-T), and the other was double mutations ( C42-T and A54- G). By statistical analysis, C42-T (encoding Ser83-keu substitution) was shown to have correlation with nalidixic-acid resistance in the clinical strains of Shigella, while A54-G (encoding Asp87-Gly substitution) was shown to have correlations with both norfloxacin resistance and ciprofloxacin resistance.
文摘According to the World Health Organization, foodborne diseases are a major public health problem, particularly in developing countries including the Republic of Congo. They are responsible for several episodes of diarrhea, especially in children under five years old. There is no reliable epidemiological data on the pathogenicity of the Shigella spp. strains circulating in the whole Republic of Congo drafting this paper. The purpose of this study was to examine the Shigella spp strain pathogenicity close to an environment contaminated with faeces in the city of Brazzaville. As a result, 54 isolates have been associated with Shigella spp. The gastric acid resistance test performed on Shigella Environmental Strain (SES) and Shigella Clinical Strains (SCS) resulted in 38.8% (21/54) and 100% acid resistant, respectively. Shigella spp. Strains (SES and SCS) were ranged in a survival percentage from 11% to 93%. By monitoring Biosurfactant-Like Molecule (BLM) production, we showed that the BLM production of SES and SCS was highly dependent on bacterial culture density involving the Quorum Sensing (QS). S. flexneri, S. boydii, and S. sonnei and as well as SES and SCS were able to invade and contaminate eggs by colonizing egg yolk. The counting bacteria were ranging from two to 5 × 10<sup>7</sup> CFU/mL after contamination. Concomitantly, BLM was secreted during the post contamination of poultry eggs with 100% EI24. Further by trying to show the pathogenicity by the hemolysis test, we have shown that SES and SCS were able to show significant areas of lysis on blood agar. Finally, this work has proposed an additional model of cell invasion including biosurfactants during the pre- and post-invasion phases.