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Objective:To evaluate the effects of Pasteurella multocida (P. multocida) vaccines on the expression and release of antibodies, interleukin (IL)-6 and IL-12 by serum.Methods:Balb/c mice were immunized with two formalin and iron inactivated vaccine doses within 2 weeks. The vaccines were adjuvant with P. multocida A strain, P. multocida B strain and Salmonella typhimurium bacterial DNA (AbDNA, BbDNA and SbDNA for short, respectively). The animals were challenged 4 weeks after immunization. Blood of mice was collected to detect the change of specific antibody, IL-6, and IL-12 using ELISA.Results:The specific antibody and interleukins in the immunized group increased significantly compared to the control mice after vaccination and challenge (P<0.05). The highest release of these cytokines was obtained by P. multocida inactivated with iron and adjuvant with AbDNA at a concentration of 25 μg/mL. The antibody titer peak was 0.447 in mice vaccinated with iron-killed whole-cell antigen adjunct with AbDNA. The time-courses of release showed that bacterial DNA was able to stimulate IL-6 and IL-12 production more than alum (P<0.05).Conclusions:Our findings introduce that bacterial DNA is capable of releasing an immunological response with several cytokines. These indicate that bacterial DNA entrapped with killed P. multocida antigen is a new and effective adjuvant to enhance specific immunity and resistance of animal against the infectious pathogen, which could simplify the development of highly promising strong adjuvant.
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Objective: To evaluate the effects of Pasteurella multocida (P. multocida) vaccines on the expression and release of antibodies, interleukin (IL)-6 and IL-12 by serum. Methods: Balb/c mice were immunized with two formalin and iron inactivated vaccine doses within 2 weeks. The vaccines were adjuvant with P. multocida A strain, P. multocida B strain and Salmonella typhimurium bacterial DNA (AbDNA, BbDNA and SbDNA for short, respectively). The animals were challenged 4 weeks after immunization. Blood of mice was collected to detect the change of specific antibody, IL-6, and IL-12 using ELISA. Results: The specific antibody and interleukins in the immunized group increased significantly compared to the control mice after vaccination and challenge (P<0.05). The highest release of these cytokines was obtained by P. multocida inactivated with iron and adjuvant with AbDNA at a concentration of 25 μg/mL. The antibody titer peak was 0.447 in mice vaccinated with iron-killed whole-cell antigen adjunct with AbDNA. The time-courses of release showed that bacterial DNA was able to stimulate IL-6 and IL-12 production more than alum (P<0.05). Conclusions: Our findings introduce that bacterial DNA is capable of releasing an immunological response with several cytokines. These indicate that bacterial DNA entrapped with killed P. multocida antigen is a new and effective adjuvant to enhance specific immunity and resistance of animal against the infectious pathogen, which could simplify the development of highly promising strong adjuvant.
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Background: Integrons are mobile genetic elements able to obtain the antibiotic resistance gene cassettes. The prevalence of integrons in the Enterobacteriaceae family has been varied and played an important role in the development of the drug resistant bacteria. The present study aimed to investigate the contribution of class 2 and 3 integrons in drug resistant Diarrheagenic Escherichia coli strains
Materials and Methods: The 164 Diarrheagenic E. coli collected from feces samples of children in the Yasuj -Iran and all isolates were identified by standard biochemical tests. The antimicrobial susceptibility for 14 antibiotics, which are used conventionally was determined by disk diffusion. The presence of class 2 and 3 integrons in all isolates was investigated by PCR
Results: Of 164 E. coli isolates from children, 80.49% carried class 2 integron and the length of the amplicons ranged from 800 bp to 2 kb. Class 3 integrons were identified among 24 E. coli isolates. All the E.coli isolates were susceptible to imipenem and the greatest resistance was correspondent to nalidixic acid. A significant correlation was revealed between Class 2 integron and resistance to kanamycin, amikacin, gentamicin, ceftazidime, chloramphenicol and cephalexin. The presence of class 3 integron was significantly associated with resistance to ampicillin, gentamicin, streptomycin, kanamycin, tetracycline and trimetoprime-sulfametoxazol
Conclusion: The results indicated that integrons are widespread in Diarrheagenic E. coli and its carriage contributed significantly to the emergence of resistance among Diarrheagenic E. coli. However, factors leading to the wide spread of integrons are still to be determined
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Background: Cervical cancer is one of the important reasons of mortality among females. Prevention, early diagnosis and immediate treatment can affect the rate of mortality in this cancer and several epidemiological studies have shown a strong relationship between human papilloma viruses [HPVs] and cervical cancer
Objectives: The present study was conducted to survey HPV infections in a women population with cervical cancer and cervical dysplasia/metaplasia in southwest of Iran
Materials and Methods: 72 paraffin-embedded cervical biopsies which had been previously archived from women with cervical cancer and cervical dysplasia were examined by polymerase chain reaction [PCR]. Afterward, the detected HPV strains were typed by restriction fragment length polymorphism [RFLP] analysis of PCR amplicons
Results: 60 out of 72 samples had necessary requirements and HPV DNA was detected in 43.3% of these samples. Most HPV positive samples belonged to women aged from 48 to 63 years. On the other hand, HPV infection among patients with squamous cell carcinoma [SCC] was 48.78% and in women with dysplasia/metaplasia was 26.66%. The most prevalent type of the human papilloma virus was HPV16 [100%]
Conclusions: Knowing the most prevalent type of the human papilloma viruses circulating in the population [HPV16] can be applied in the future screening and managing programs of this major disease and also in vaccination against the prevalent types of the virus. Meanwhile, it seems that more studies should be performed to determine the role of different risk factors involved in development of the disease, especially those related with social behaviors and traditions with respect to different areas
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OBJECTIVE@#To survey the prevalence severe diarrhea arising from these bacteria in children under 5 years old in Marvdasht.@*METHODS@#In this study faecal sample from 615 children aged <5 years old who were hospitalized for gastroenteritis in Fars hospitals in Iran were collected and then enriched in Escherichia coli (E. coli) broth and modified tryptone soy broth with novobiocin media. Fermentation of sorbitol, lactose and β -glucoronidase activity of isolated strains was examined by CT-SMAC, VRBA and chromogenic media respectively. Then isolation of E. coli O157:H7 have been confirmed with the use of specific antisera and with multiplex PCR method presence of virulence genes including: stx1, stx2, eaeA, hly has been analyzed.@*RESULTS@#E. coli O157:H7 was detected in 7 (1.14%) stool specimens. A significant difference was seen between detection rate of isolated bacteria from age groups 18-23 months and other age groups (P=0.004). Out of considered virulence genes, only 1 of the isolated strains (0.16%) the stx1 and eaeA genes were seen and also all isolated bacteria had resistance to penicillin, ampicillin and erythromycin antibiotics.@*CONCLUSIONS@#We found that children < 2 years of age were at highest risk of infection with E. coli O157:H7. Regarding severity of E. coli O157:H7 pathogenesis, low infectious dose and lack of routine assay for detection of these bacteria in clinical laboratory, further and completed studies on diagnosis and genotyping of this E. coli O157:H7 strain has been recommended.
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Objective:To survey the prevalence severe diarrhea arising from these bacteria in children under5 years old inMarvdasht.Methods:In this study faecal sample from615 children aged <5 years old who were hospitalized for gastroenteritis inFars hospitals inIran were collected and then enriched inEscherichia coli(E. coli) broth and modified tryptone soy broth with novobiocin media.Fermentation of sorbitol, lactose and β-glucoronidase activity of isolated strains was examined byCT-SMAC,VRBA and chromogenic media respectively.Then isolation ofE. coli O157:H7 have been confirmed with the use of specific antisera and with multiplexPCR method presence of virulence genes including: stx1,stx2,eaeA,hly has been analyzed.Results:E. coli O157:H7 was detected in7(1.14%) stool specimens.A significant difference was seen between detection rate of isolated bacteria from age groups18-23 months and other age groups(P=0.004). Out of considered virulence genes, only1 of the isolated strains(0.16%) the stx1 andeaeA genes were seen and also all isolated bacteria had resistance to penicillin, ampicillin and erythromycin antibiotics.Conclusions:We found that children <2 years of age were at highest risk of infection withE. coliO157:H7.Regarding severity ofE. coliO157:H7 pathogenesis, low infectious dose and lack of routine assay for detection of these bacteria in clinical laboratory, further and completed studies on diagnosis and genotyping of thisE. coliO157:H7 strain has been recommended.
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Objective: To survey the prevalence severe diarrhea arising from these bacteria in children under 5 years old in Marvdasht. Methods: In this study faecal sample from 615 children aged 1, stx2, eaeA, hly has been analyzed. Results: E. coli O157:H7 was detected in 7 (1.14%) stool specimens. A significant difference was seen between detection rate of isolated bacteria from age groups 18-23 months and other age groups (P=0.004). Out of considered virulence genes, only 1 of the isolated strains (0.16%) the stx1 and eaeA genes were seen and also all isolated bacteria had resistance to penicillin, ampicillin and erythromycin antibiotics. Conclusions: We found that children < 2 years of age were at highest risk of infection with E. coli O157:H7. Regarding severity of E. coli O157:H7 pathogenesis, low infectious dose and lack of routine assay for detection of these bacteria in clinical laboratory, further and completed studies on diagnosis and genotyping of this E. coli O157:H7 strain has been recommended.
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OBJECTIVE@#To identify three common genes (blaTEM, blaSHV and blaCTX-M) responsible for ESBL production in Klebsiella pneumoniae (K. pneumoniae)isolated from Intensive Care Units of Namazi Hospital, Shiraz, Iran.@*METHODS@#A total of 60 non-repetitive nosocomial isolates from 60 patients were selected during 2009-2010. The phenotypic identification of ESBL production was confirmed by Double Disk Synergy Test (DDST) according to CLSI guidelines. The ESBL's genotype was then analyzed by multiplex PCR of blaTEM, blaSHV and blaCTX-M genes and DNA sequencing.@*RESULTS@#The primary susceptibility tests of K. pneumoniae showed that among 10 examined antibiotics, the most resistant and susceptible antibiotics identified in this study were ampicillin and imipenem, respectively. The phenotypic determination of ESBL by DDST showed that 60% (n=36) of isolates produced ESBL. Multiplex PCR of genes among K. pneumoniae isolates showed that 39% (n=18) of them have TEM, 39% (n=18) of them have both CTX-M and TEM and 13% (n=8) of them have TEM, SHV, CTX-M.@*CONCLUSIONS@#Our findings reveal the high prevalence (60%) of ESBL producing K. pneumoniae from ICU patients along with a new pattern of blaTEMdistribution differ from other countries.
Subject(s)
Female , Humans , Male , Anti-Bacterial Agents , Pharmacology , Bacterial Proteins , Genetics , Metabolism , Cross Infection , Epidemiology , Microbiology , Intensive Care Units , Iran , Klebsiella Infections , Epidemiology , Microbiology , Klebsiella pneumoniae , Genetics , Multiplex Polymerase Chain Reaction , Methods , Prevalence , Referral and Consultation , beta-Lactamases , Genetics , MetabolismABSTRACT
Calcium oxalate is one the most significant causes of human kidney stones. Increasing oxalate uptake results in increased urinary oxalate. Elevated urinary oxalate is one the most important causes of kidney stone formation. This study aims to evaluate oxalate-degrading capacity of lactic acid bacteria and its impact on incidence of kidney stone. This case-control study was conducted on serum, urinary, and fecal samples. The research population included a total of 200 subjects divided in two equal groups. They were selected from the patients with urinary tract stones, visiting urologist, and also normal people. The level of calcium, oxalate, and citrate in the urinary samples, parathyroid and calcium in the serum samples, and degrading activity of fecal lactobacillus strains of all the subjects were evaluated. Then, data analysis was carried out using SPSS-11.5, chi[2] test, Fisher's exact test, and analysis of variance. The results revealed that the patients had higher urinary level of oxalate and calcium, as well as higher serum level of parathyroid hormone than normal people. In contrast, urinary level of citrate was higher in normal people. In addition, there was a significant difference between the oxalate-degrading capacities of lactobacillus isolated from the patients and their normal peers. Reduction of digestive lactobacillus-related oxalate-degrading capacity and increased serum level of parathyroid hormone can cause elevated urinary level of oxalate and calcium in people with kidney stone
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Rotaviruses are one of the most important causes of acute gastroenteritis and death in children worldwide. WHO suggested hospital based surveillance all over the world in order to evaluate the prevalence of rotavirus diarrhea. This study was conducted to determine the prevalence of rotavirus gastroenteritis in hospitalized children in Shiraz. In this study 138 stool samples from children aged <5 years old with severe diarrhea [>/= 3 loose watery stools per 24 hours], hospitalized at Shaheed Dastgheib and Nemazee hospitals in Shiraz, were collected during 2006-2007. All the stool specimens were evaluated for Group A of rotaviruses with enzyme immunoassays [EIA]. Then demographic and clinical data were analyzed by SPSS software. Out of total collected samples rotavirus infection was detected in 48[34.78%]. The highest infection rate was among children less than two years old [70.83%]. Diarrhea [97.92%], vomiting [77.08%] and fever [52.08%] were the most frequent reported clinical symptoms in children with rotavirus infection. The highest of isolation of virus was observed in autumn [45.83%] and the lowest in spring [8.33%] [p=0.012]. Also, there was no significant difference between the frequency of the rotavirus diarrhea and the pattern of nutrition [p= 0.236]. Regarding to high frequency of rotavirus infection, concurrently surveillance of rotavirus gastroenteritis in other hospitals of Iran is recommended.
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The group A human rotavirus is the most prevalent agents causing diarrhea in children which is found in water sources including waste and treatment waters worldwide. The aim of this study was environmental surveillance of group A human Rotavirus from influent and effluent of urban and hospital sewage systems in Shiraz. In this cross-sectional study, 60 specimens from both influent and effluent system of urban and hospital sewage disposal systems of Shiraz Nemazee hospital were collected by using grab sampling method. All samples concentrated by using two concentration methods, including Pellet and Two-phase. Then Group A human rotaviruses were detected with enzyme immunoassay [EIA]. 15 [25%] group A human rotaviruses were detected, of which 11 samples [73/33%] belonged to the influent part and 4 samples [26/67%] were related to the effluent. There was a significant difference between influent and effluent of systems. Also statistical significant differences were found between the distribution of rotaviruses and months of sampling. This study reveals the inefficiency of refining system in attempting to complete elimination of group A human rotaviruses.
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This study was conducted to evaluate the prevalence of rotavirus disease and to investigate the genotypes of rotavirus strains causing acute gastroenteritis among children aged <5 years old in Marvdasht, Iran. One hundred and forty-one children, aged 1 month to 5 years, afflicted with severe diarrhea were enrolled during January 2007 to December 2008. Their stool samples were studied with enzyme immunoassays [EIA] for group A rotaviruses. Rotavirus-positive specimens were genotyped by the Nested RT-PCR using different types of specific primers. Out of total collected samples rotavirus infection was detected in 40 [28.37%]. Of the rotavirus episodes, 72.91% occurred during the first 2 years of life [P=0.038]. The highest prevalence of infection was identified in summer [52.50%] and the lowest in winter [7.50%]. The most common clinical features included diarrhea [96.25%], vomiting [82.50%] and fever [45.0%]. Mixed genotypes were the predominant G type [60.0%], followed by non-typeable [12.50%], G2 [12.50%], G4 [10.0%] and G1 [5.0%] genotypes. G3/G8 mixed infection is the first of these rotavirus genotypes to be reported in Iran. Regarding high frequency of rotavirus infection, continuous surveillance is needed to inform diarrhea prevention programs as well as to provide information about the occurrence of new rotavirus strains. This will assist policy makers in decision making on rotavirus vaccine introduction
Subject(s)
Humans , Male , Female , Rotavirus Infections/epidemiology , Child , Gastroenteritis/virology , Molecular Epidemiology , Diarrhea , Immunoenzyme Techniques , Reverse Transcriptase Polymerase Chain Reaction , Vomiting , Fever , GenotypeABSTRACT
Helicobacter pylori [H. pylori] has several virulence factors such as vacA and cagA genes. Mosaicism in vacA alleles with two distinct families of vacA signal sequences [s1, s2] and two distinct families of middle region alleles [m1, m2] is reported. The aim of this study was determination of H. pylori vacA allelic types in Chaharmahal and Bakhtiyari province, Iran and their correlation with six different gastroduodenal diseases. This cross-sectional descriptive study was performed on 200 antral gastric biopsy specimens that were obtained from patients undergoing upper gastrointestinal tract endoscopy in Hajar Hospital of Shahrekord. Initially, H. pylori strains were identified by rapid urease test [RUT] and ureC primers, and thereafter, we used seven specific primers for detection of vacA genotypes. Statistical analyses were used to find their relationship to gastric disorders. The frequency of the vacA alleles, s1a/m1a, s1a/m1b, s1a/m2, s1b/m1a, s1b/m1b, s1b/m2, s1c/m1a, s1c/m1b, s1c/m2, s2/m1a, s2/m1b and s2/m2 were respectively, 27[16.46%], 8[4.87%], 45[28.43%], 7[4.26%], 5[3.04%], 10[6.09%], 12[7.31%], 4[2.43%], 18[10.97%], 6[3.65%], 0 and 22[13.41%]. s1a/m2 strains were the most prevalent strains in this region and there was a considerable relationship between s1a/m1a, s1a/m2, s2/m2 and s1c/m1a with some gastric disorders. As the findings are different from other regions of the world, extended molecular epidemiologic investigations are recommended in other cities of Iran
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Helicobactr pylori [H.pylori] infection is related to chronic gastritis, peptic ulcer and duodenal ulcer. Thus, identification and treatment of the infection has a considerable importance. The aim of this study was to compare three methods of polymerase chain reaction [PCR], culture and rapid urease test [RUT] in identification of Helicobactr pylori in gastric biopsy specimens. 263 patients [157 woman and 106 man] who suffering from digestive complaints and referred to the endoscopy department of Hajar Hospital in Sharkord [2007, Iran] were participated in the study. Three gastric biopsy samples were collected from each patient. Samples were examined by standard RUT and culture methods for diagnosis of H.pylori. PCR was used for diagnosis of ureC gene. Out of 263 patients, 36.49%, 9.50%, 13.30%, 9.48% and 5.50% had criteria gastritis, gastric ulcer, duodenal ulcer, esophagitis and gastric cancer, respectively. H.pylori infection was diagnosed in 54.37%, 31.94% and 84.79% of the patients by RUT, culture and PCR method, respectively. Findings of this study indicated that PCR has a greater sensitivity rather than rapid urease test and culture methods for diagnosis of H.pylori
Subject(s)
Humans , Male , Female , Helicobacter Infections/diagnosis , Polymerase Chain Reaction , Urease , Culture Techniques , Stomach/pathology , Biopsy , Gastritis , Stomach Ulcer , Duodenal Ulcer , Esophagitis , Stomach NeoplasmsABSTRACT
Enterohemorrhagic Escherichia coli [EHEC] strains are one of the most important enteric pathogens which cause of the food-borne disease in humans. The purpose of this study was to survey the prevalence of diarrhea arising from this bacteria and infection source and antibiotic resistance of isolated strains in children under 5 years old in Marvdasht. In this descriptive study, Stool samples of under 5 years old children in Marvdasht were collected. After enrichment in specific culture media and evaluation sorbitol fermentation and their p-glucoronidase activity [MUG test] with specific antisera, the isolation of EHEC O157:H7 strain was confirmed. Then, virulence genes verotoxin, intimin and hemolysin with multiplex PCR and antibiotic resistance strains with disk diffusion method were tested. Also, getting infection with water, food and animal contact was evaluated. Out of 615 cases, 89 sorbitol negative colonies isolated which 7 strains O157 [1.14%] confirmed with antisera. Of virulence genes, only 1 of the isolated strains [0.16%] had stxj and eaeA genes. All isolated bacteria had resistance to penicillin, ampicillin and erythromycin. There was no significant relation between isolated EHEC strains and the different seasons of the year. Hospital based surveillance to estimate the burden of EHEC strains in clinical centers in all less than 5 years old children with acute gastroenteritis and studies on etiology and genotyping of this bacterium in other parts of the country are recommended
Subject(s)
Humans , Child, Preschool , Drug Resistance, Microbial , Prevalence , Feces/microbiology , Polymerase Chain Reaction , Health SurveysABSTRACT
Listeria monocytogenes is the causative agent of listeriosis, a highly fatal opportunistic food borne infection. Listeriolysin O is a major virulence factor in this bacterium, which is encoded by hlyA. The aim of the present study was to determine the relation of Listeria monocytogenes hlyA gene isolated from fresh cheese with habitual abortion in Marvdasht. In this cross-sectional study, 428 fresh cheese samples from four geographical area of Marvdasht were collected, then cold enriched and cultured in Hicrome Listeria Agar and Palcam Agar. Specific biochemical and sugar fermentation tests were used for identification of probability bacteria. Finally, hlyA gene was determined by PCR method. Of 428 samples, 56 [13.1%] L. monocytogenes were isolated, among which 91.7% were revealed to encode hlyA. Data analysis revealed significant association between months of sampling and isolated bacteria [p<0.004]. Also, there was a significant association between bacteria and hly A gene with human abortion. Fresh cheese and unpasteurized milk contaminated with L.monocytogenesis can be one of the reasons of habitual abortion in Marvdasht. Therefore pregnant women and immunocompromised people are strongly recommended not to consume fresh cheese and unpasteurized milk
Subject(s)
Cheese , Abortion, Habitual , Listeriosis , Bacterial Toxins , Cross-Sectional Studies , Polymerase Chain ReactionABSTRACT
Tuberculosis [TB] caused by Mycobacterium tuberculosis, is an infectious disease in human which kills nearly three millions of people annually. Approximately, one - third of the world populations are infected with this bacteria and 5 - 10% of them develop the active form of the disease. Individuals are different in susceptibility to TB infection. These differences might be due to the host characteristics especially genetic factors. TNF- alpha as a pro-inflammatory cytokine, plays a key role in host defense against tuberculosis. Presence of mutation in this gene can influence the effectiveness, performance and capability of immune responses against TB infection. The Aim of this study was to investigate the frequency of TNF- alpha gene polymorphisms and its relation with susceptibility to the pulmonary TB. Sixty healthy controls and 60 TB patients were enrolled. Genotype of TNF[-238], TNF -244, TNF[-308], TNF[-857] and TNF[-863] were determined using PCR-RFLP method. The results were analyzed by Fisher Exact and kappa[2] tests using SPSS v.14 and evaluated with Hardy-Weinberg equilibrium. The results of this study showed a significant difference in TNF-308 and TNF [-857] regions between the control and study groups [P < 0.05]. Presence of mutation in TNF[-308] and TNF [-857] regions may increase the host susceptibility to mycobacterium tuberculosis and genotyping of these regions can be used for screening of the high risk individuals
Subject(s)
Humans , Mycobacterium tuberculosis , Tumor Necrosis Factor-alpha , Polymorphism, Genetic , Polymerase Chain ReactionABSTRACT
Metronidazole is one of the most important antibiotics in Helicobacter pylori [H.pylori] eradication regimens. This study was designed to determine metronidazole resistance H. pylori isolates from patients referred to Hajar hospital, Shahrekord, Iran. In this cross-sectional study, 263 patients who referred to endoscopy department of Hajar hospital, Shahrekord in 2007 were randomly selected. Gastric biopsy samples were cultured on selective Brucella agar containing 10% blood and incubated under microerophilic condition at 37°C between 3 and 7 days. Gram stain, urease, catalase and oxidase tests were used to detect H. Pylori isolates, and PCR [polymerase chain reaction] to determine ureC gene. H.pylori indentified. Standard CLSI [Clinical and Laboratory Standard Institute] was used to evaluate metronidazole resistance. Rapid urease test [RUT], culture and PCR detect H. pylori infection in 54.37%, 84% and 84.79%, respectively. Of isolated strains, 49 [58.33%] metronidazole resistance and 7 [8.33%] semisensitive were recognized. Of 49 metronidazole resistant patients, 32.65%, 20.40%, 12.24%, 6.10%, 4.80% and 24.48% had gastritis, gastric ulcer, duodenal ulcer, esophagitis, gastric cancer and normal endoscopy, respectively. Fisher exact test shows that there are relation between metronidazole resistance and women. Regarding high metronidazole resistance among studied patients, alternative antibiotics with less resistance is recommended in Helicobacter pylori [H.pylori] eradication regimens