Evaluation of cross immunity and histopathological findings in experimentally infected BALB/c mice with neospora caninum and besnoitia caprae

Caprine besnoitiosis is an economically important disease of goats. Neospora caninum, another coccidian parasite of worldwide distribution, infects several animal species and is a major cause of abortion in cattle. Combined infections of N. caninum and Besnoitia caprae can occur in geographical areas endemic for both species of parasite in goats. This experiment was conducted to investigate the possible cross-immunity between these two infections in experimentally infected BALB/c mice. Forty BALB/c mice were divided into four equal groups. The mice of Groups 1 and 4 were inoculated with 1x10[6] live virulent tachyzoites of N. caninum [NC-1], while animals of Groups 2 and 3 were inoculated with sterile tissue culture medium. Each mouse in Groups 1 and 2 was challenged 28 days later with 1 xl0[6] live virulent bradyzoites of-B. Caprae [BC-1]. Following the challenge, the mice in Groups 1 and 2 showed 100% morbidity and 100% mortality within 9 days post infection, while all the animals of Groups 3 and 4 remained alive. The dead animals were necropsied. The survivors [mice in Group 3 and 4] were euthanized 9 days after inoculation and the gross and histopathological lesions in different organs were investigated. Immunization and challenge experiments with lethal dose of B. caprae in the highly susceptible BALB/c mice showed no cross-protection between N. caninum and B. caprae

[Identification of enterotoxigenic Escherichia coli strains in children under 2 years of age by real-time PCR in Shiraz]

Enterotoxigenic strains of E. coli [ETEC] form the second largest group of diarrheagenic E. coli and are responsible for more than 25% of infantile diseases. The purpose of this study is to assess the prevalence of ETEC strains and their virulence genes in children less than 2 years of age in Shiraz. This was a cross-sectional study. 285 stool samples were taken from children under 2 years of age with diarrhea in Shiraz. First, E. coli strains were isolated using standard biochemical tests. Then, prevalence of ETEC strains and presence of st and lt genes were evaluated using real-time PCR. Antibiotic susceptibility of the isolates was evaluated by using disc diffusion method. 49 [17%] diarrheagenic E. coli strains were isolated, 7 cases [14/3%] belonged to ETEC strain. Among isolated ETEC strains, 4 [57.14%] had lt, 1 [14.29%] had stIa and 2 strains [28.57%] had both lt and stIa genes. All ETEC strains were susceptible to chloramphenicol, amikacin and nitrofurantoin, but showed resistance to penicillin and macrolides. The results showed high prevalence of ETEC strains in our study area. We recommend hospital-wide surveillance by using molecular techniques to estimate the prevalence of this pathotype in other regions of our country

An investigation on As, Cd, Mo and Cu contents of soils surrounding the Meyduk tailings dam

The contents of As, Cd, Cu and Mo were determined in soil surrounding Meyduk tailings dam based on 21 surface soil samples. Assessment of toxic element pollution in studied soil samples needs knowledge of pre-anthropogenic metal concentration to act as a reference against which measured values could be compared. Estimating the background values was executed by sampling rocks adjacent to soil sampling stations. Various indices including enrichment factor [EF], pollution load index [PLI], modified contamination degree [mCd] and geoaccumulation index [I geo] were used for determining the contamination level of soil in the vicinity of tailings impoundment under the effect of the tailings dust. Anthropogenic pollution was diagnosed from natural one by sequential extraction done by Tessier et al method and calculating pollution intensity index [l poli]. The results indicate a significant upward enrichment in northeastern and southwestern surface soil around the tailings dam for Cd and Cu. The dominant wind direction demonstrated that only the contamination of southwestern soil around the tailings dam would be attributed to tailings dust while the dispersive dusts scattered after the mine explosions generally affected the northeastern part

[Evaluation of molecular mechanisms resistance to macrolide by S. pneumoniae strains isolated from Nemazee and Shahid Faghihi Hospitals in Shiraz]

Macrolide Streptococcus pneumoniae is generally mediated by two mechanisms: 23SrRNA methylation and efflux. The aim of this study was to assess the molecular mechanism of resistance in S.pneumoniae strains isolated from intensive care units in Shiraz. This was a cross-sectional study. 50 strains of S.pneumoniae were obtained from the patients admitted to intensive care units [ICU] of Nemazee and Shahid-Faghihi Hospitals, from 2010 to 2011. Suspected colonies were identified by phenotypical and biochemical tests. Organisms were confirmed to be Streptococcus pneumoniae on the basis of the presence of lytA gene by PCR method. Antibiotic resistance was evaluated according to Standard Clinical and Laboratory Standards Institute [CLSI]. Macrolide resistance genes were identified by use of ermB and mefA specific primers. Using SPSS software, statistical analysis was performed by means of chi-square test. Resistance to erythromycin, clarithromycin and azithromycin levofloxacin were 18%, 48% and 44% respectively. In this study, 25 strains [50%] had mefA and 21 [42%] had ermB genes. Erythromycin resistance had significant relationships with mefA and ermB genes. The results of our study showed that the predominant mechanism of macrolide resistance was due to mefA gene in this area. Increased macrolide resistance calls for attention to the pattern of resistance in the therapeutic regimens

[Epidemiological features of rotaviral, bacterial, and parasitic infections among hospitalized children in Jahrom [2006-2007]]

Rotavirus is one of the most common cause of diarrhea and one of the major causes of severe gastroenteritis in very young children. To follow up and genotype the agents of rotavirus infection as well as assessing the bacterial and parasitic organisms among hospitalized children with gastroenteritis in the city of Jahrom, Iran. This cross-sectional descriptive study was carried out during October 2006 to October 2007. A total of 163 stool samples from hospitalized children less than 5 years old with severe diarrhea were collected from two hospitals in Jahrom. Culture, microscopy, EIA, and RT-PCR were used for detection of bacterial, parasitic and rotaviral agents. Data were analyzed using SPSS 14 and descriptive statistics including chi-square test, ANOVA, and Fisher exact test. A p value less than 0.05 was considered to be statistically significant. Of total samples, 46.02% were positive for group A rotavirus by EIA. The predominant genotypes were G[1] [17.33%], G[4] [30.66%], and nontypable [30.66%]. Also, E.coli, Shigella spp., Shigella spp. + E.coli, E.coli + rotavirus, Salmonella spp., E. histolytica/E.Dispar, and other infectious agents were identified in 7.97%, 17.18%, 1.83%, 15.20%, 3.66%, 10.84%, and 6.28% of cases, respectively. According, to the data obtained from the present study, rotavirus infections in Jahrom mostly occur within the cold months of the year, epidemiologically

Association between TNF-alpha [-857] gene polymorphism and susceptibility to tuberculosis

TNF-alpha as a pro-inflammatory cytokine play a key role in host defense against tuberculosis [TB]. Presence of mutation in TNF-alpha gene can influence the effectiveness, performance and capability of immune responses against this infection. The Aim of this study was to investigate the frequency of TNF-alpha alleles and its relationship with susceptibility to TB and TNF-alpha gene variations. A case-control study was conducted and 103 healthy controls and 93 TB patients were enrolled. Genotype of TNF-238, TNF -244, TNF-308, TNF -857 and TNF-863 were distinguished using PCR-RFLP method. TNF-857 and TNF-863 were in high frequency mutation regions in a population level, and a significant difference at TNF-857 was noticed between the two groups of case and control. Presence of mutation in TNF-857 region probably increases the host susceptibility to mycobacterial infection. Genotyping of these regions in combination with other factors can be used for screening of high risk persons. According to high distribution of mutations in TNF-857 and TNF-863 regions, further studies on association of these regions is suggested

[Molecular and serological characterization of group A rotavirus isolates obtained from hospitalized children in Jahrom]

Rotaviruses are the most common factors causing gastroenteritis in children. Considering the high rate of mortality in developing countries, WHO proposed hospital based surveillance in order to identify the prevalent genotype profile. This study was conducted to determine the prevalent genotypes of Rotavirus by RT-PCR method in hospitalized children in Jahrom city. In this study, 163 stool samples were collected from children [<5 years old] with sever diarrhea who were hospitalized in two hospitals in Jahrom city during 2005-2006. At first, group A rotavirus were distinguished by Enzyme Immunoassay [EIA] test and then the genotypes of positive samples were determined using 9 specific primers by Nested RT-PCR method. From the total of 163 collected samples, 75 samples were positive by ELISA. The frequency of G1,G2,G3,G4,G9 and the mix genotypes were 17.33%, 13.34%, 2.67%, 30.66%, 2.67% and 2.67%, respectively. The most common prevalence of positive rotavirus was observed in winter with 22.69% compared to the least prevalence in summer with 4.29% [P<0.05]. Considering the high prevalence of positive rotavirus in studied population and also prevalence of rare genotypes, we suggest a broader research in other parts of the country

Comparison of Integrated Cell Culture -RT-PCR and Cell Culture Methods for detection of Enteroviruses

Generally, sewage exposed water could be potentially contaminated with enteroviruses. For this reason, enterovirus isolation from sewage specimens is one of the most sensitive indicators for virus circulation in the population. We evaluated the ICC-RT-PCR and cell culture methods for detection of enteroviruses in Tehran sewage system. This research utilized 63 specimens provided through Grab sample method to concentrate by two-phase method and cultured in RD and HEp-2 cells, respectively. All specimens then were inoculated using sensitive cell cultures of RD and HEp-2. After 24 hours incubation at 36[degree sign] by means of Pan E.V primers and afterwards Pan P.V Primers along with specific sabin primers, RT-PCR was carried out on the cell culture specimens. Data were analyzed using SPSS Software [SPSS for and ANOVA test as well as Chi-square test. Out of 63 collected specimens, enteroviruses were isolated from 33 specimens [52.38%] and 41[65.01%] specimens which utilized cell-culture and ICC-RT-PCR methods respectively. Polioviruses were also isolated from 6 specimens. Statistical analysis indicated that there was a significant relationship [0.05 level] between cell culture and ICC-RT-PCR methods to isolate enteroviruses. Further the sensitivity of ICC-RT-PCR method to detect enteroviruses less than 0.01 TCID 50 was evaluated, which indicated that this method is acceptable and sensitive enough to detect enteroviruses in sewage

Study on the histopathological and enzymatic relationship in chickens affected by Marek's Disease

The aim of the present study was to determine the relationship between histopathological lesions and tissue enzymes in chickens affected by Marek's disease.Five apparently healthy chickens [as control group] and 25 chickens affected by Marek's disease were selected. After consideration of history and gross lesions, tissue samples were collected from kidney, liver, heart, ovary, pectoral muscle, spleen, crop, proventriculus and bursa of fabricius for histopathological and tissue enzyme studies. In tissue samples, the activity of aspartate aminotransferase [AST], alkaline phosphatase [ALP], lactate dehydrogenase [LDH], creatine kinase [CK], superoxide dismutase [SOD] and glutathione peroxidase [GPX]was measured. Histopathologically, lymphomatous lesions in visceral organs were included in local or diffuse ple omorphic small to medium lymphocytes, lymphoblasts, marek cells and rarely plasma cells. The results indicated that the activities of AST, LDH, CK, SOD and GPX were decreased in the crop of affected chickens. The activities of CK, LDH and AST in the liver tumors and the activities of LDH, AST, ALP, SOD, CK and GPX in pectoral muscles were increased. In proventriculus, the activities of AST, ALP, LDH and SOD were shown to be increased, but the activity of CK was decreased. The activities of AST and SOD in the ovary lesions and the activities of SOD, AST, CK and LDH in the heart lesions showed an increase. Fluctuations in the activity of enzymes in different tissues showed that measurement of these enzymes can not be used as a tumor marker in the diagnosis of Marek's disease. In order to determine whether these enzymes are tumor markers or not, the measurement of the specific isoenzymes of each enzyme is necessary

Environmental surveillance of Polio and Non-Polio Enteroviruses in Sistan and Balouchestan Province

Enteroviruses can easily circulate in the population through sewage and they are suitable indicators for environmental surveillance. On the other hand, in some countries there are evidences of silent circulation of viruses in sewage specimens despite no virus isolation from clinical specimens. Therefore, WHO has suggested environmental surveillance using surface water and sewage specimens for final confirmation of Poliovirus eradication. In this research, according to wild Poliovirus circulation in Afghanistan and Pakistan and probability of virus entrance to Iran, and also to assure wild Poliovirus eradication, the environmental surveillance was performed in Sistan and Balouchestan Province of Iran. From March 2004 to February 2005, 86 specimens from 2 sewage disposal systems, 5 hospitals and surface water from several villages were collected by Grab Sample method and tested for Enteroviruses directly and using 2 concentration methods: Pellet and Two-phase. Then Poliovirus and Non-Polio Enteroviruses [NPEV] were serotyped by microneutralization method and Polioviruses were intratypically differentiated using ELISA and Probe Hybridization techniques. From a total of 86 specimens, Enteroviruses and Non-Polio Enteroviruses were isolated from 49[56.98%] and 46[53.49%] of specimens respectively. Polioviruses were isolated from 18[20.93%] specimens and none of them was wild Poliovirus fortunately. 13[17.81%], 39[53.42%] and 57[78.08%] of enteroviruses were isolated using Direct, Pellet and Two-phase methods, respectively. The results of this research confirm the validity of environmental surveillance and Polio eradication in Sistan and Balouchestan Province

[Study on epidemiological patterns of Mycobacterium tuberculosis by fingerprinting]

Despite the availability of effective anti tubercle chemotherapy for more than 50 years and major advances in the biology of M. tuberculosis, TB remains the leading cause of adult mortality attributable to a single pathogen. The analysis of tuberculosis and tracing of the source of infection require the ability to discriminate among Mycobacterium tuberculosis strain. In present study Restriction Fragment Length Polymorphism [RFLP] analysis was used to study the molecular epidemiology of tuberculosis in Tehran. Mycobacterium tuberculosis isolates from 292 patients with culture positive tuberculosis during 2002-2003. Extraction of bacterial DNA and DNA fingerprinting with RFLP using lS6110 as probe, was performed by standard protocols. The digested DNA is separated according to fragment size on agarose gel by electrophoresis and southern transferred on to a membrane. The DNA probe was labeled with [HRP] and hybridizing to the genomic targets. Among 292 culture M. tuberculosis, 232 [79.4%] belonged to clusters and 60 [21.6%] did not. 39 Drugs resistant M. tuberculosis isolates were examined 33%of these were identical pattern lS6110. 4.9% of the isolates represented the Beijing genotype. Based on the obtained data it appears that tuberculosis among the study population in Tehran mainly from reactivation of latent infection

[Assessment of different methods to remove organic inhibitors in order to create a sensitive method for direct surveillance of enteroviral infectious diseases in sewage specimens]

Enteroviruses in sewage are considered among the most sensitive indicators for virus circulation in society. These are mainly detected by sensitive cell cultures, however, since it is time consuming, molecular direct methods have also been considered as sensitive techniques. This study accomplished to assess different methods of removing organic inhibitors of sewage in order to detect Enteroviruses with RT-PCR. For this cross sectional study, 63 sewage specimens of Tehran were prepared with Grab sample method and concentrated with Pellet and Two-Phase methods and cultured in RD and Hep-2 cells. Then, with 12 different methods, removing organic inhibitors in sewage with RT-PCR method was assessed. Of investigated methods, we succeeded to isolate all of expected viruses only with the use of ICC-RT-PCR method. With respect to 0.01 TCID50 sensitivity of ICC-RT-PCR method, further studies are strongly recommended in order to confirm the utility of this method as one of the most sensitive methods of detecting Enteroviruses

[Study on the prevalence of urinary tract infection by Escherichia coli, antibiotic resistance and plasmid profile of isolated bacteria in Jahrom city]

Background: Urinary tract infection is one of the most common bacterial infections, that creates in outpatient and hospitalized patients. Since, E. coli is the most common bacteria in UTI infection, the aim of this study was to determine urinary tract infections by Escherichia coli, antibiotic resistance and plasmid profile detection of isolated E. coli in Jahrom city

Materials and methods: This investigation was carried out for 6 months on patients with urinary tract infection at Peimaniye hospital and Dr. jazayeri private laboratory Jahrom city in 2004. Totally, 3361 of urine samples were collected. Bacteria were isolated and identified by bacterial standard methods. Then, tests were carried out on 100 selected of E. coli randomly. Antimicrobial resistance tests were done by disk diffusion agar. The bacterial plasmid also was extracted by alkalin lysis and detected by electrophoresis through agarose gels

Results: Bacterial cultures of 356 samples were positive. Isolated bacteria were E. coli [80.34%], Klebsiella [10.67%], Enterobacter [3.65%], Citrobacter [1.69%], Pseudomonas [1.41%], Staphylococci [0.84%], Serracia [0.84%] and Proteus [0.56%]. E. coli isolates were shown the most resistance to cotrimoxazole [49%] and cephalexin [30%] and ciprofloxacin was known as the most effective antibiotic with 82% sensitivity. Eighty percent of bacteria had plasmids. The most plasmid bands were detected in isolated bacteria of hospitalized patients. All bacteria that were resistant to at least one kind of investingated antibiotics, had plasmids

Conclusions: Plasmid profile similarity of isolated bacteria from hospitalized patients, indicated probability these bacteria originate from one bacterial clone or high prevalence gene transmition among hospitalized bacteria. So, antibiogram test for each patient, infection control and limitation of using antibiotics can prevent spreading of resistant bacteria

Study on the prevalence of urinary tract infection by Escherichia coli, antibiotic resistance and plasmid profile of isolated bacteria in Jahrom city

Urinary tract infection is one of the most common bacterial infections, that creates in outpatient and hospitalized patients. Since, E. coli is the most common bacteria in UTI infection, the aim of this study was to determine urinary tract infections by Escherichia coli, antibiotic resistance and plasmid profile detection of isolated E. coli in Jahrom city. This investigation was carried out for 6 months on patients with urinary tract infection at Peimaniye hospital and Dr. jazayeri private laboratory Jahrom city in 2004. Totally, 3361 of urine samples were collected. Bacteria were isolated and identified by bacterial standard methods. Then, tests were carried out on 100 selected of E. coli randomly. Antimicrobial resistance tests were done by disk diffusion agar. The bacterial plasmid also was extracted by alkalin lysis and detected by electrophoresis through agarose gels. Bacterial cultures of 356 samples were positive. Isolated bacteria were E. coli [80.34%], Klebsiella [10.67%], Enterobacter [3.65%], Citrobacter [1.69%], Pseudomonas [1.41%], Staphylococci [0.84%], Serracia [0.84%] and Proteus [0.56%]. E. coli isolates were shown the most resistance to cotrimoxazole [49%] and cephalexin [30%] and ciprofloxacin was known as the most effective antibiotic with 82% sensitivity. Eighty percent of bacteria had plasmids. The most plasmid bands were detected in isolated bacteria of hospitalized patients All bacteria that were resistant to at least one kind of investigated antibiotics, had plasmids. Plasmid profile similarity of isolated bacteria from hospitalized patients, indicated probability these bacteria originate from one bacterial clone or high prevalence gene transmition among hospitalized bacteria. So, antibiogram test for each patient, infection control and limitation of using antibiotics can prevent spreading of resistant bacteria.

Poliovirus environmental surveillance in sistan-balouchestan province, isolation and identification of viruses by cell culture and microneutralization, and intratypic differentiation by ELISA and probe hybridization

In some countries, wild polioviruses have been isolated from environment despite the absence of viruses being recovered from clinical cases, therefore to confirm of final Polio eradication, WHO has recommended environmental surveillance using sewage specimens and surface water. During the present study, in order to assure the polio eradication in Iran, Sistan-Balouchestan province was chosen as the target area. During a 12-month period, 86 specimens from 2 sewage disposal systems and 5 hospitals, as well as surface water from several rural areas were collected by Grab Sampling and tested for polioviruses using direct and concentrated specimens with 2 concentration methods: Pellet and Two-phase. Then the isolated viruses were serotyped by microneutralization method and differentiated intratypically by ELISA and probe hybridization techniques. Of all studied specimens, 18 [20.9%] were identified as poliovirus, none of which were wild virus, fortunately. Among these, 2 [2.3%], 8 [9.3%] and 13 [15.1%] were isolated from direct specimens, Pellet and Two-phase concentrated specimens, respectively. The most frequent viruses were Polio 2 [72.2%] and Polio 3 [27.8%]. Results have revealed the efficacy of immunization coverage in Iran. Meanwhile, sufficient surveillance programs have been observed during the recent years