Jundishapur Journal of Microbiology
Volume:8 Issue: 3, Mar 2015

Merkel cell carcinoma (MCC) is a rare and highly aggressive malignancy of the skin which occurs mainly in old people and is very uncommon in young individuals. A new tumor virus belonging to the Polyomaviridae family; Merkel Cell Polyomavirus (MCPyV) has recently been identified in more than 80% of MCCs. We conducted a retrospective review on the archives of the Department of Pathology; Imam Khomeini Hospital Cancer Institute affiliated to Tehran University of Medical Sciences to confirm the MCC samples and we found medical records and samples of a young case with MCC who developed leg skin and scalp tumor six and seven years after bone marrow transplantation, respectively. We analyzed patient formalin-fixed paraffin-embedded samples for the presence of MCPyV DNA using polymerase chain reaction (PCR) method, and the PCR amplicons were subjected to DNA sequencing. Merkel Cell Polyomavirus DNA was detected in both tumors from patient and sequence analysis of the viral LT3 region showed a close homology to strains circulating worldwide. The findings of this study are consistent with the hypothesis that local, systemic, or tumor-induced immunosuppression may allow the MCPyV to initiate skin aggressive cancer. It is necessary to maintain regular check over patients taking immunosuppressive medications for MCPyV infection. Since there is not any information about detection and molecular biology analysis of MCPyV among Iranian patients with MCC, this study provides more information about MCC and MCPyV in Iran.

Listeria monocytogenes is a significant zoonosis causing invasive infections in the susceptible persons. The current paper presented a patient who died due to a rapidly-progressing multiple organ failure (MOF) as a result of severe sepsis caused by L. monocytogenes. A 70-years-old patient with chronic renal failure was admitted to the infectious diseases clinic due to diarrhea for one day. He was hospitalized and the body fluid samples were collected for laboratory analyses. Within few hours, his vital findings worsened, and he developed respiratory arrest. Ceftriaxone and gentamycin were administrated. However, he died due to disseminated intravascular coagulation, septic shock and meningoencephalitis at the 22nd hour of admission. Causative agent was identified as L. monocytogenes serotype-4b in post-mortem period. L. monocytogenes can cause progressive and rapidly fatal infections in the vulnerable persons, with multisystem involvement. Since this bacterium is not susceptible to cephalosporines, it will be better to consider effective antimicrobials in the treatment of the possible cases.

Bovine viral diarrhea (BVD) is an economically important disease of cattle distributed worldwide. Diagnosis of BVD relies on laboratory-based detection of its viral causing agent or virus specific antibodies and the most common laboratory method for this purpose is Enzyme-Linked Immunosorbent Assay (ELISA). The current study was aimed to develop a simple indirect ELISA to detect antibodies against Bovine Viral Diarrhea Virus (BVDV) in the sera of infected cattle. A new simple indirect ELISA method was developed to detect BVDV infection by prokaryotically (Escherichia coli, BL21 strain) expressed recombinant whole nonstructural protein 3 (NS3) of BVDV (NADL strain). Four hundred bovine serum samples were evaluated by the newly developed NS3-ELISA and virus neutralization test (VNT) as the gold standard method to diagnose BVD. Among these samples, 289 sera had been previously tested by a commercial ELISA kit. Statistical analyses showed a very high correlation between the results of the developed NS3-ELISA and VNT (kappa coefficient = 0.935, P < 0.001), with the relative sensitivity and specificity of 94% and 98.8%, respectively. There was also a high correlation between the results of NS3-ELISA and the commercial ELISA kit (kappa coefficient = 0.802, P < 0.001) with the relative sensitivity and specificity of 90.72% and 91.15%, respectively. The newly developed simple indirect ELISA showed high sensitivity and specificity with respect to VNT. Developing such a simple, sensitive, and specific ELISA which is much less expensive than the available commercial ELISA kits can improve the detection of BVDV infections, help to eliminate the disease from herds, and decrease economic losses caused by this disease.

Blastocystis is a zoonotic protozoan parasite living in the digestive system of some vertebrates. This parasite has some subtypes, pathogenicity status of which has still remained controversial. The aim of this study was to determine the subtype of Blastocystis in infected cattle. This descriptive cross-sectional study was performed on 196 isolates from cattle stool samples collected from slaughterhouse in Khorramabad city, Iran, in 2012. Genomic DNA was extracted and to determine the Blastocystis subtype, seven pairs of sequence-tagged sites (STS) primers were used. Of 196 specimens, 19 (9.6%) were infected with Blastocystis. Among the 19 positive samples, the most common subtype was ST5 (47.36 %), followed by ST3 (10.53%) and ST6 (10.53%). Two (10.53%) samples had mixed infections by ST3 and ST5. The four isolates not amplified by any STS primers were probably unknown genotypes. In the present study, the highest prevalence was for ST5, which is so important for epidemiology and risk of human infection. The report related to ST3 in cattle as a subtype of human showed mutual infection between human and cattle. Another important point in this study was the ST6 report. Finally, it seems that gathering epidemiological data is needed for a better understanding of the potential animal reservoirs for human infection.

Neisseria lactamica as one of the main commensal in oropharynx during the childhood is related to the induction of a natural immunity against meningococcal meningitis. Also Moraxella catarrhalis in oropharynx of children is a predisposing factor for otitis media infection. The current study aimed to investigate the frequency of the N. lactamica, other nonpathogenic Neisseria spp. and M. catarrhalis in the oropharynx of young healthy children in Ahvaz, Iran by the two phenotypic tests and Polymerase Chain Reaction (PCR). A total of 192 oropharyngeal swab samples of the young healthy children were studied during four months. Swabs were plated onto enriched selective media and non-selective media. Gram-negative and oxidase-positive diplococci were identified by several conventional biochemical tests. The PCR and sequencing were used to confirm the accuracy of laboratory diagnosis to identify N. lactamica and M. catarrhalis. Among 192 young healthy children with the mean age of 5.93 ± 2.5903 years, authors identified: N. lactamica (21.9%) in the age group of one to nine years; N. mucosa (6.3%); N. sicca (7.8%); N. cinerea (1.6%); N. subflava (biovar subflava) (4.2%); N. subflava (biovar perflava) (28.1%); N. subflava (biovar flava) (7.3%) and M. catarrhalis (42.7%). The young healthy children screening by colonization of N. lactamica and other nonpathogenic Neisseria spp. in oropharynx was the first report in Ahvaz, Iran. The study results demonstrated the high frequency of colonization of M. catarrhalis in the studied young healthy children other than Neisseria spp.

Hepatitis B virus (HBV) infection is a worldwide public health problem. Nine HBV genotypes (A-I) have been already discovered. HBV genotypes are important both in the clinical manifestation of disease and treatment response. Moreover, HBV DNA without HBs (Hepatitis B surface)-antigenemia was detected in some patients with chronic hepatitis (occult hepatitis). There is little information about HBV genotypes and its relation to occult infection despite the importance of this infection in Khuzestan Province. This study aimed to determine both occult hepatitis B infection and HBV genotypes among cirrhotic patients.Patients and Thirty-eight patients with liver cirrhosis, including 11 (28.9%) HBsAg-positive patients and 27 (71.1%) patients with cryptogenic cirrhosis participated in this study. The mean age of the patients at the time of cirrhosis diagnosis was 54.85 years (range 26-75 years). All patients were anti-HCV and anti-HIV negative. For all the samples, the serological Enzyme-Linked Immunosorbent Assay (ELISA) was performed for HBV markers including HBsAg, HBcAb, HBeAg, HBeAb tests. The common primer of S region of HBV was used for Nested PCR. The PCR products of the positive individuals were sequenced for genotyping and subtyping of HBV. Eleven (40.7%) out of 27 HBV cryptogenic cirrhosis and all 11 HBsAg-positive patients were positive for HBV DNA. The seroprevalences of Hepatitis B virus HBe antigen, anti-HBe and anti-HBc antibodies among the cryptogenic cirrhosis patients were 5 (18.5%), 1 (3.7%), and 5 (20.83), and among HBsAg-positive patients were 6 (54.5%), 5 (45.5%), and 7 (63.6%), respectively. In our study, only HBV genotype D was found among all the positive HBsAg and occult HBV infection. Moreover, high prevalence (40.7%) of occult HBV infection was determined among patients suffered from cryptogenic cirrhosis.

Biological control of parasitic nematodes by microorganisms is a promising approach to control such parasites. Microorganisms such as fungi, viruses and bacteria are recognized as biocontrol agents of nematodes. The current study mainly aimed to evaluate the in vitro Potential of various saprophyte soil-fungi in reducing the infective larvae stage of parasitic nematode Trichostrongylidae family. Sheep feces were employed to provide the required third stage larvae source for the experiments. The nematode infective larvae of Trichostrongylidae family including three species of Ostertagia circumcincta, Marshalgia marshali and Heamonchos contortus were collected by Berman apparatus. Fifteen isolates of filamentous fungi were tested in the current study. One milliliter suspension containing 200 third stage larvae of Trichostrongylidae family was separately added to the fungal cultures in 2% water-agar medium Petri-dishes. Every day the live larvae were counted with light microscope (10X) and the number of captured larvae was recorded on different days. Significant differences were observed in the results of co-culture of nematodes larva and fungi after seven days. The most effective fungi against the nematodes larvae were Cladosporium sp., Trichoderma sp., Fusarium equisetti, after seven days of incubation. The studies on fungi could be applied as suitable tools in biocontrol of nematode infections. However, additional surveys are required to select efficient with the ability to reduce the nematode larvae in the environment.

Pseudomonas aeruginosa is an important nosocomial pathogen characterized by its innate resistance to multiple antimicrobial agents. Plasmid-mediated drug resistance also occurs by the production of extended-spectrum β-lactamases (ESBL), metallo β-lactamases (MBL), and AmpC β-lactamases. Another important factor for establishment of chronic infections by P. aeruginosa is biofilm formation mediated by the psl gene cluster. The aim of this study was to evaluate biofilm formation and presence of the pslA gene in burn isolates of P. aeruginosa as well as the association of antibiotic resistance, MBL, ESBL and AmpC β-lactamase production with biofilm formation among the isolates. Sixty-two burn isolates of P. aeruginosa were obtained from Shahid Motahari Hospital in Tehran from August to October 2011. Antibiotic susceptibility was determined by the disc diffusion assay. MBL, AmpC and ESBL production were screened using the double disc synergy test, AmpC disc test and combined disc diffusion assay, respectively. The potential to form biofilm was measured using the microtiter plate assay and pslA gene was detected using specific primers and PCR. Biofilm formation was observed in 43.5% of the isolates, of which 66.7% produced strong and 33.3% formed weak biofilms. All biofilm-positive and 14.2% of biofilm-negative isolates harbored the pslA gene. MBL, AmpC and ESBL production were significantly higher in the biofilm-positive isolates (70.3%, 62.9% and 33.3%, respectively) compared to the biofilm-negative strains (31.4%, 34.2% and 20%, respectively). Overall, 19 isolates (30.6%) co-produced MBL and AmpC, among which the majority were biofilm-positive (63.1%). Finally, four isolates (6.4%) had all three enzymes, of which 3 (75%) produced biofilm. Biofilm formation (both strong and weak) strongly correlated with pslA gene carriage. Biofilm formation also correlated with MBL and AmpC β-lactamase production. More importantly, multiple-β-lactamase phenotype was associated with formation of strong biofilms.

Because of the economic importance of the Arab race horses and also the role of Toxoplasma gondii and Neospora spp. in abortion and reproductive failure of these animals, we decided to perform this study. We designed this study to investigate the seroprevalence of anti-Toxoplasma gondii and anti-Neospora spp. antibodies in Arab horses from 12 cities of Khuzestan province in southwest of Iran. From October 2009 to March 2011, a total of 235 blood samples were collected from jugular veins of Arab horses of different ages and genders from 12 cities of Khuzestan province. All the sera were tested for anti-Toxoplasma antibodies using the modified agglutination test (MAT) and the existence of anti-Neospora antibodies were tested using N-MAT for Neospora spp. According to the MAT results, antibodies to T. gondii were found in 114 (48.5%) of 235 sera with titers of 1:20 in 84, 1:40 in 19, 1:80 in four, 1:160 in four, and 1:320 in three horses. According to the N-MAT results, antibodies to Neospora spp. were found in 47 (20%) of 235 sera with titers of 1:40 in 39, 1:80 in five, and 1:160 in three horses. We did not observe any statistically significant differences regarding age groups and genders between seropositive and seronegative horses for Neospora spp. using chi-square (χ2) test, but it seemed that anti-Toxoplasma antibodies were more prevalent in older horses (≥ 10 years old). The results indicated that Arab horses are exposed to these parasites in southwest of Iran. Further research is required to determine the genomic structures of these parasites in Arab horses in southwest of Iran.

Globally, almost 20% of cancers are related to infectious agents that can be prevented. Oncogenicity refers to viruses that may cause cancers, more importantly in immunocompromised subjects such as transplant and hemodialysis patients. Therefore, epidemiological studies are the first line for understanding the importance of these agents in public health, particularly, in mobile populations, tourism and pilgrimage regions. Oncogenic viral infections, such as hepatitis B virus (HBV), hepatitis C virus (HCV) and Epstein-barr virus (EBV) are the most common viral agents in immunocompromised patients. Furthermore, human T lymphocyte virus type I (HTLV-I), due to endemicity in Khorasan Razavi province located northeast of Iran as a pilgrimage region, and Kaposi's sarcoma associated herpes virus (KSHV), as an oncogenic herpesvirus in immunocompromised subjects have been investigated among the general population and those with end-stage renal diseases (ESRD).Patients and A cross-sectional study was carried out among 1227 randomly selected individuals; 25 donors and 195 patients with ESRD, including 60 kidney transplant recipients and 135 dialysis patients from the Khorasan Razavi province, Iran. Serological tests were carried out using commercial enzyme-immunoassay kits. To confirm positive serology tests, the extracted viral DNA or RNA was examined for the presence of KSHV, HTLV-I and HCV by conventional PCR. The prevalence of KSHV infection in the general population was 1.71% (21/1227); 2.60% (10/384) males and 1.30% (11/843) females. In kidney transplants, viral infections occurred in 23.3% of subjects; including EBV, HTLV-I and HBV-HCV co-infection in 8.3%, 3.3% and 1.7%, respectively. In patients on hemodialysis, viral infections were present in 29.6% including EBV, HTLV-I and HBV-HCV co-infection in 2.2%, 5.9% and 16.3%, respectively. Seroprevalence of KSHV in patients with kidney transplants was 1.7% and in patients on dialysis was 3.0%. Furthermore, KSHV and HTLV-I genome was detected in 25% and 100% of seropositive subjects, respectively. In conclusion, this study demonstrated that these tumor virus infections including HTLV-I, KSHV and particularly hepatitis viruses (HBV plus HCV) are prevalent in the general population and in patients on hemodialysis, which might be an important health concern in this region due to the mobile population.

Phage therapy or use of lytic bacteriophages for eliminating bacterial populations has been developed for several aspects of human affairs such as medicine, agriculture and food industries. The high load of coliforms of treated wastewater effluents that are discharged into the rivers or agricultural lands is a serious concern of the Iran Department of Environment and the reduction of coliforms using phages to overcome this problem is an asset. This research aimed to isolate and identify specific lytic coliphages and investigate their effects on native and standard Escherichia coli strains as well as coliform populations in municipal wastewater. The wastewater sample was cultured on selective culture media to isolate a native coliform strain and characterized using molecular methods. River water was centrifuged and passed through a 0.45 μm filter and its lytic coliphages were enriched and purified against a native E. coli as well as a standard E. coli strain. Municipal wastewater was treated with isolated lytic coliphages and most probable number (MPN) reduction was examined. E. coli SBSWF27, which is a native strain of E. coli from Isfahan municipal wastewater treatment plant, was isolated and characterized. Also two novel bacteriophages related to Myoviridae and Podoviridae families of bacteriophages from Zayandehrood River (Isfahan, Iran) were isolated. These coliphages had lytic effects on E. coli PTCC1399 and E. coli SBSWF27 as coliform's index. The myovirus had a hexagonal head measuring 27.28 nm and a noncontractile tail measuring 204.5 × 13.63 nm. The podovirus had an oval head measuring 98 × 35 nm and a tail, 14 nm in diameter. The treatment of municipal sewage with the coliphage mixture resulted in a 22-fold decrease of the coliform's MPN from 2400 to 110 after two hours of incubation. This is the first report on isolation and identification of two novel lytic myovirus and podovirus from Zayandehrood River in Isfahan that had lytic effects on E. coli PTCC1399 and E. coli SBSWF27 strains as well as coliform's population of Isfahan municipal wastewater. We suggest that the use of these lytic coliphages for reduction of coliform's population in sewage could be considered as an effective and simple alternative for costly replacement of instruments and establishments of the old wastewater treatment plants.

Legionellaceae contains Legionella genus with over 52 species and 64 serogroups. It is one of the most important causes of respiratory disease in human. More than 30% of hospital-acquired pneumonia is caused by Legionella. Ventilator-associated pneumonia (VAP) is an infection acquired in hospital wards, particularly in intensive care unit (ICU). This disease approximately affects 9% to 20% of intubated patients. Mortality in these patients varies between 8% and 76%. Legionella is one of the important factors for infection in intubated patients. The present study was aimed to investigate the use of molecular methods in diagnosis of infection caused by Legionella pneumophila. In this study, 109 samples of lung secretions collected from intubated patients admitted to ICU wards of four university hospitals in a three-month period were examined. Cultivation and Real time Polymerase Chain Reaction (PCR) methods were used to assess L. pneumophila colonization in these samples. Eleven samples had positive results using real time PCR analysis of 16s rRNA gene fragments specific for L. pneumophila, but according to culture method on specific buffered charcoal-yeast extract medium (BCYE), no positive cases were detected. Of the total positive cases, six were males, one female and four infants. The seven adults aged 40-65 years. Using molecular methods in diagnosis of infection caused by L. pneumophila has a great value because of its high specificity and rapid diagnosis potency.

Viruses are one of the major reasons of gastrointestinal disease worldwide, and commonly infect children less than five years of age in developing countries. The current study aimed to determine the frequency of adenoviruses, rotaviruses and noroviruses among diarrhea samples collected from infants of Zabol, south-east of Iran. This study is the first investigation of adenoviruses, rotaviruses and noroviruses among diarrhea samples in Zabol.Patients and In this study, eighty-two diarrhea stool samples were collected from infants aged < 1 to 12 months admitted to the hospital, internal laboratory and central laboratory of Zabol, Iran. All samples were subjected to the rapid immunochromatography assay. The results showed that the frequency of rotaviruses, adenoviruses and noroviruses among infants with diarrhea were 70.20%, 20.30% and 9.50%, respectively. There were 50 subjects aged one to five months and 2 subjects aged nine to twelve months. The results of geographical distribution showed that the number of infants living in rural and urban areas with these viruses were 50 and 32, respectively. Rotaviruses were most common in rural and urban infants with 42 and 10 cases, respectively. Regarding the feeding patterns of infants with diarrhea, mixed feeding and breast feeding were found in 51 and 31 cases, respectively. In conclusion, the results of our study showed that the major viral pathogens that caused infantile diarrhea in Zabol city were rotaviruses followed by adenoviruses and noroviruses. The results of our study can useful for prosperous control of infantile diarrhea.

Halomethanes are toxic and carcinogenic chemicals, which are widely used in industry. Also they can be formed during water disinfection by chlorine. Biodegradation by methylotrophs is the most important way to remove these pollutants from the environment. This study aimed to represent a simple and rapid method for quantitative study of halomethanes utilizing bacteria in drinking water and also a method to facilitate the biodegradation of these compounds in the environment compared to cometabolism. Enumeration of chlorinated methane utilizing bacteria in drinking water was carried out by most probable number (MPN) method in two steps. First, the presence and the number of methylotroph bacteria were confirmed on methanol-containing medium. Then, utilization of dichloromethane was determined by measuring the released chloride after the addition of 0.04 mol/L of it to the growth medium. Also, the effect of nanosilver particles on biodegradation of multiple chlorinated methanes was studied by bacterial growth on Bushnell-Haas Broth containing chloroform (trichloromethane) that was treated with 0.2 ppm nanosilver. Most probable number of methylotrophs and chlorinated methane utilizing bacteria in tested drinking water were 10 and 4 MPN Index/L, respectively. Chloroform treatment by nanosilver leads to dechlorination and the production of formaldehyde. The highest growth of bacteria and formic acid production were observed in the tubes containing 1% chloroform treated with nanosilver. By combining the two tests, a rapid approach to estimation of most probable number of chlorinated methane utilizing bacteria is introduced. Treatment by nanosilver particles was resulted in the easier and faster biodegradation of chloroform by bacteria. Thus, degradation of these chlorinated compounds is more efficient compared to cometabolism.

Although there is enough evidence that infectious agents such as Chlamydia pneumonia and Helicobacter pylori may play a pathogenic role in atherosclerosis, this role for cytomegalovirus (CMV) is yet controversial. The aim of the present study was to detect CMV-DNA in atherosclerotic plaques in patients who underwent coronary artery bypass graft (CABG).Patients and In this case-control study, candidates for CABG (cases) and patients with valvular or congenital malformation but without atherosclerotic plaques (controls) were studied from 2012 to 2013 at Golestan hospital, Ahvaz, IR Iran. Demographic and laboratory data were collected. Atherosclerotic and histological samples were obtained from visible plaques and from aorta by the surgeon. All the samples were examined for the presence of CMV-DNA by polymerase chain reaction (PCR) method using a commercial kit (SinaClon, Tehran, IR Iran). The mean ages in case and control groups were 60.8 ± 6.8 and 57.5 ± 11.5 years, respectively, with no significant difference (P = 0.09). Thirty patients (54.5%) in case and 32 (58.2%) in control groups were male with no significant difference (P = 0.7). CMV-DNA was present in 8 (14.5%) of the cases and 2 (4%) of the controls. CMV-DNA was associated with higher risk of atherosclerosis (OR: 7.7, 95% CI = 1.1-51.4, P = 0.03). Of the total normal aortic samples (55 in cases and 55 in controls), there was no individual with simultaneous positive CMV-DNA among aortic atherosclerotic and normal tissue samples. The presence of CMV-DNA in aortic plaques is associated with increased risk of atherosclerosis. CMV infection may be considered as an independent risk factor for this event.

Boletus griseipurpureus Corner, an edible mushroom, is a putative ectomycorrhizal fungus. Currently, the taxonomic boundary of this mushroom is unclear and its bitter taste makes it interesting for evaluating its antibacterial properties. The purpose of this study was to identify the genetic variation of this mushroom and also to evaluate any antibacterial activities. Basidiocarps were collected from 2 north-eastern provinces, Roi Et and Ubon Ratchathani, and from 2 southern provinces, Songkhla and Surat Thani, in Thailand. Genomic DNA was extracted and molecular structure was examined using the RNA polymerase II (RPB2) analysis. Antibacterial activities of basidiocarp extracts were conducted with Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 29523 and methicillin-resistant Staphylococcus aureus (MRSA) 189 using the agar-well diffusion method. All the samples collected for this study constituted a monophyletic clade, which was closely related with the Boletus group of polypore fungi. For the antibacterial study, it was found that the crude methanol extract of basidiomes inhibited the growth of all bacteria in vitro more than the crude ethyl acetate extract. Basidomes collected from four locations in Thailand had low genetic variation and their extracts inhibited the growth of all tested bacteria. The health benefits of this edible species should be evaluated further.

Diagnosis of tuberculosis (TB) is not always easy, thus employing methods with a short duration and acceptable sensitivity and specificity is necessary to diagnose TB. The aim of this study was to investigate the diagnostic value of serum adenosine deaminase (ADA) level for diagnosis of pulmonary tuberculosis.Patients and A total of 160 sex and age-matched subjects were included in this study, and were divided to four groups; forty patients with pulmonary tuberculosis (PTB) diagnosed based on the national TB program (NTP), forty patients with non-tuberculosis bacterial pneumonia, forty patients with lung cancer and forty people who were healthy in every respect. Serum adenosine deaminase activity in patients of each group was measured by the Giusti and Galanti calorimetry method using a commercial kit (Diazyme, USA). The ANOVA analysis was used to compare groups for quantitative variables. Mean serum ADA level in the PTB group was clearly higher than the mean serum ADA in the other three groups. Mean serum ADA was 26 IU/L in PTB patients, 19.48 IU/L in patients with pneumonia, 15.8 IU/L in patients with lung cancer, and 10.7 IU/L in the control group (P < 0.05). In regard to the cut off value of 26 IU/L for ADA in patients with PTB sensitivity and specificity was defined as 35% and 91%, respectively. Serum ADA activity with high specificity percentage may be a useful alternative test in restricted resource areas to rule out diagnosis of PTB. However, serum ADA activity is not a useful tool for TB diagnosis.

Early treatment of pulmonary tuberculosis (PTB) is necessary for a successful tuberculosis (TB) control program. The objective of this study was to determine total treatment delay and its associated factors among PTB patients in Ahvaz.Patients and A retrospective study was performed among newly diagnosed PTB cases registered in 2010 at the Ahvaz health center. Total treatment delay was defined as the time interval between the onsets of cough to the initiation of anti-TB treatment. Tuberculosis diagnosis and treatment was based on the national TB program (NTP). Data analysis was performed using the SPSS software by chi-square and Fisher's exact test with odds ratio (OR) and 95% confidence interval (CI). The mean age of the patients was 38.9 ± 12.3 years; 83 were male and 56 were female. Of the 139 smear positive PTB cases, 91 (65.5%) cases had received delayed-treatment. The mean time between onset of symptoms, diagnosis and treatment was 73 days (median: 48 days, range: 4-570 days). Female gender (OR (95% CI): 2.9, 1.03-8.23, P = 0.02), smoking (OR (95% CI): 0.49, 0.22-0.96, P = 0.04) and receiving immunosuppressive drugs (OR (95% CI): 8.18, 1.09-75.31, P < 0.05) were associated with longer delayed time. Delayed diagnosis and treatment of tuberculosis appears to be the main problem in the TB control program of the region. Delayed time is significantly associated with female gender, smoking and immunosuppressive drugs.

BK virus (BKV) belongs to the human Polyomaviridae and the primary BKV infection is occurred during childhood then the virus could be latent through life, especially in the kidneys and urinary system. It became reactive after an immunocompromised status, such as pregnancy or transplantation. Isolated BKV from different locations of the world is grouped into four subtypes using serological and genotyping methods. The BKV subtype I is the dominant one and has worldwide distribution. According to our knowledge, there are no data about the BKV prevalence and its genotypes in southwest part of Iran. Considering the high prevalence of renal failure and kidney transplant patients in this part, and the role of BKV in graft rejection, this study aimed to determine the prevalence of BKV infection in renal transplant recipients referred to Golestan Hospital in Ahvaz City, Iran.Patients and Urine samples were collected from 122 kidney transplant recipients referred to Golestan Hospital in Ahvaz, southwest of Iran. The extracted DNA was amplified by Polymerase Chain Reaction, and subtype of each positive sample was determined using Restriction Fragment Length Polymorphism (RFLP) and sequencing methods. From all study population, 51/122 (41.8%) urine samples were positive for BKV DNA and the other samples were negative (71/122). Forty-eight cases (94.11%) were subtype I and 3 others (5.89%) were subtype IV using the RFLP method. None of the patient’s urine samples were positive for subtypes II and III. Our work is the second study in Iran and considering huge numbers of transplantation in Iran and Khuzestan Province, south western of Iran, in addition to the role of this virus in kidney transplant rejection, routine evaluation of BKV positivity is recommended both for graft recipient and donors. This helps better transplantation result and may prevent graft rejection.

Based on many studies, otitis media with effusion (OME) is one of the major causes of childhood hearing loss, social malformation and medical costs. The pathogenesis still remains unclear, though it is known that this complication is closely related to bacterial infections. Alloiococcus otitidis, Haemophilus influenzae, Streptococcus pneumoniae and Moraxella catarrhalis are the most common bacterial pathogens isolated from middle ear effusions (MEEs). Due to the prevalence of OME in children, we decided to investigate bacterial agents that cause diseases such as A. otitidis, H. influenzae, S. pneumonia and M. catarrhalis in these subjects.Patients and Forty-five children between one and 15 years of age were selected for this study. Seventy specimens were collected from MEE by myringotomy and inoculated in PBS buffer. Conventional culture and PCR methods were used for identification of bacterial agents. The bacterial cultures in 8.6% of samples were positive by conventional culture, with A. otitidis, M. catarrhalis and S. pneumoniae present in 1.4%, 2.9% and 4.3% of samples, respectively. No H. influenzae was isolated. By the PCR method, A. otitidis was the most frequently isolated bacterium, found in 25.7% of samples, followed by S. pneumoniae, M. catarrhalis and H. influenzae, which were identified in 20%, 12% and 20% of samples, respectively. Overall, 55 out of 70 samples were positive by both the PCR and culture method. It can be concluded that A. otitidis was the major causative agent of MEE in children with OME. Therefore clinicians should be aware that bacterial infection plays an important role in the progression of acute otitis media to OME in children of our region.

Hepatitis B virus (HBV) is a major cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma (HCC). HB vaccination is an essential step in the prevention of the disease and its consequent complications. Immune status of medical personnel in teaching hospitals of Khuzestan is not well known. Since these personnel are usually at risk of needle stick and other high-risk events, some challenges exist in infection control committee with regard to managing these events. This study was conducted to assess post-vaccination immunity status and non-response to HBV vaccine as well as its predictors among medical staff in a teaching hospital affiliated to Ahvaz Jundishapur University of Medical Sciences (AJUMS) in Ahvaz, southwest Iran.Patients and In this retrospective cross-sectional study, the medical staff of a teaching hospital was evaluated for their immune response against HBV and factors affecting it. The study conducted in Razi Hospital, Ahvaz City, southwest of Iran, in 2013. Demographical, clinical, and laboratory data registered in medical files of hospital staff were analyzed by SPSS software version 16 using chi-square and Fisher exact tests. Differences with P value < 0.05 were considered significant. To identify predictors for non-responders, we used odds ratio (OR) with 95% confidence interval (CI). Out of 239 participants, 43 (18 %) were men and 196 (82%) were women. Their mean age was 31.9 ± 18.1 years (range of 20 - 55 years). Fourteen (5.9%) participants were non-responders, 37 (15.5%) were poor responders, and 188 (78.6%) were good responders. The non-responders were older (> 50 years) than the responders (P = 0.0001), while the body mass index (BMI) was not significantly different (P = 0.37) between them. Diabetes mellitus (DM) (OR: 7.3, 95% CI, 1.3 - 41.7, P = 0.05), and using immunosuppressive drugs (ISD) (OR: 3.2, 95% CI, 1.1 - 11.5, P = 0.03) were two variables in association with non-response to HB vaccine. Non-response rate to HB vaccine in our study was approximately 6%. Age over 50 years, DM, and receiving immunosuppressive drugs may be considered as predictors for non-response to HB vaccine in medical staff.

Chronic hepatitis B virus (HBV) infection has a broad spectrum of manifestation, ranging from silent carrier state to advanced cirrhosis and hepatocellular carcinoma. The persistence of HBV DNA in serum and hepatocytes of the cirrhotic patient could be detected by molecular techniques in spite of negative HBV serologic markers. This case-control study was designed to evaluate the prevalence of occult HBV infection (OBI) in patients with cryptogenic liver cirrhosis in comparison with healthy subjects.Patients and Of 165 patients with liver cirrhosis, 50 consecutive patients with cryptogenic cirrhosis and 80 healthy individual without any risk factors as a control group were enrolled in this study. Their sera were tested for HBV DNA using nested PCR method. Of 50 patients with cryptogenic cirrhotic, 36 (72%) were male. The mean age of patients was 53.34 ± 14.73 years; 80 healthy subjects were selected as control group with mean age of 32.65 ± 8.51 years; 7 (14%) of the patients with cryptogenic cirrhosis showed positive HBV DNA by PCR, while HBV DNA was negative for the control group (P = 0.0001); 4(57%) cases with positive HBV shown by PCR were negative for anti-HBc and anti-HBs tests. The mean level of transaminases was significantly higher in patients with cirrhosis. There were no significant differences in demographic parameters, transaminases level and degree of hepatic failure among cirrhotic patients with and without OBI. The prevalence of OBI was relatively high in patients with cryptogenic cirrhosis. OBI was found among the patients above 40 years old. Prospective cohort studies are needed to evaluate the clinical significance of OBI.

Extracellular phospholipase, proteinase, and coagulase are accounted as the major virulence factors in Candida albicans. Several reports showed that the incidence of resistance to fluconazole has risen during last two decades. The present study has investigated the extracellular enzymes of C. albicans and non-albicans species isolated from both patients with vaginitis and healthy women. In addition, susceptibility of the isolates was evaluated against fluconazole.Patients and Vaginal samples were collected using sterile cotton swabs and inoculated on CHROMagar Candida. Routine morphological tests and ID 32C and API 20C AUX Kits were used to identify species. Phospholipase, proteinase, and coagulase activity were determined by standard methods. Susceptibility to fluconazole was also evaluated using ATB Fungus 3 Kits. The phospholipase activity was detected in 66.7% of the tested isolates recovered from patients with vaginitis. In the present study, phospholipase activity with higher Pz values (< 0.70) was more common in patients with vaginitis (28 of 66 isolates) whereas this rate in the normal individual was 13 of 42. Proteinase activity was detected in 74.2% and 61.9% of tested isolates recovered from patients and normal individuals, respectively. All tested isolates were negative for coagulase activity. In the present study, resistance to fluconazole was found in 34.8% of isolates. C. dubliniensis was the most common isolate (6 out of 11 isolates) that showed resistance to fluconazole. Our results showed that C. albicans was the most frequently isolated from both patients with vaginitis and normal individual. In the present study, we could not find any correlation between extracellular activities and sources of isolates (patients and normal flora) and sensitivity or resistance to fluconazole.

Chronic rhinosinusitis (CRS) is one of the most common chronic illnesses, but the etiology and pathogenesis of CRS are not well understood. Few studies have been carried out on the role of viruses in patients with chronic sinusitis so far. Regarding the high number of patients, we intended to evaluate the prevalence of rhinovirus and respiratory syncytial virus in patients with CRS. Doing so, we may pave the way for definitely achieving the causes and factors of the disease and consequently definite treatment of this debilitating disease in future studies.Patients and This cross-sectional study was carried out on 76 patients. Sample of the study consisted of patients with CRS who were candidates for functional endoscopic sinus surgery (FESS). The specimens were collected during FESS between February 2013 and December 2013. For this purpose, after entering into sinuses, the specimens were collected from their mucus. They were then placed in Dulbecco's modified Eagle's viral transport medium (DMEM). They were transferred to the virology lab of the university in a cold chain. The specimens were maintained in -70°C before examination. Reverse transcriptase-polymerase chain reaction (RT-PCR) was applied to explore the presence of rhinovirus and respiratory syncytial virus. In this study, 76 patients with rhinosinusitis underwent FESS as qualified candidates. The sample of the study consisted of 48 males (63.2%) and 28 females (36.8%) with the mean age of 44.3 years and an age range of 19-76 years. Among the 76 patients, 53 were with polyps and 23 without polyps. Among the patients, 66 were candidates of FESS for the first time. Other 11 patients had previously undergone the surgery. The results from PCR indicated that 22 (28.94%) patients had rhinovirus and 9 (11.84%) had respiratory syncytial virus (RSV). A total of 25 patients (32.89%) had one of the two viruses. In 6 (7.89%) specimens, both viruses were reported. CRS is a common disease with negative effects on the quality of patients’ lives. This study showed the high prevalence of these two common respiratory viruses in patients with CRS.

Toxoplasmosis is an opportunistic protozoan infection with a high prevalence in a broad range of hosts infecting up to one-third of the world human population. Toxoplasmosis leads to serious medical problems in immunocompromised individuals and fetuses and also induces abortion and mortality in domestic animals. Therefore, there is a huge demand for the development of an effective vaccine. Surface Antigen 1 (SAG1) is one of the important immunodominant surface antigens of Toxoplasma gondii, which interacts with host cells and primarily involved in adhesion, invasion and stimulation of host immune response. Surface antigen 1 is considered as the leading candidate for development of an effective vaccine against toxoplasmosis. The purpose of this study was to clone the major surface antigen1 gene (SAG1) from the genotype 1 of T. gondii, RH strain into the eukaryotic expression vector pVAX1 in order to use for a DNA vaccine. Genomic DNA was extracted from tachyzoite of the parasite using the QIAamp DNA mini kit. After designing the specific primers, SAG1 gene was amplified by Polymerase Chain Reaction (PCR). The purified PCR products were then cloned into a pPrime plasmid vector. The aforementioned product was subcloned into the pVAX1 eukaryotic expression vector. The recombinant pVAX1-SAG1 was then transfected into Chinese Hamster Ovary (CHO) cells and expression of SAG1 antigen was evaluated using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR), Immunofluorescence Assay (IFA) and Western Blotting (WB).: The cloning and subcloning products (pPrime-SAG1 and pVAX1-SAG1 plasmid vectors) of SAG1 gene were verified and confirmed by enzyme digestion and sequencing. A 30 kDa recombinant protein was expressed in CHO cells as shown by IFA and WB methods. The pVAX1 expression vector and CHO cells are a suitable system for high-level recombinant protein production for SAG1 gene from T. gondii parasites and are promising approaches for antigen preparation in vaccine development.

Diabetes mellitus as a chronic metabolic disease occurs in patients with partial or complete deficiency of insulin secretion or disorder in action of insulin on tissue. The disease is known to provide conditions for overgrowth of Candida species. Candida spp. cause candidiasis by many virulence factors such as esterase, hemolysin and phospholipase. This study aimed to compare esterase and hemolytic activity in various Candida species isolated from oral cavity of diabetic and non-diabetic individuals.Patients and Swab samples were taken from 95 patients with diabetes (35 men and 60 women) and 95 normal persons (42 men and 53 women) and cultured on Sabouraud dextrose agar. Identification of isolated yeasts was performed by germ tube test, morphology on CHROMagar Candida medium, corn meal agar and ability to grow at 45°C. Hemolysin activity was evaluated using blood plate assay and esterase activity was determined using the Tween 80 opacity test. Different Candida species were isolated from 57 (60%) diabetic and 24 (25%) non-diabetic individuals. Esterase activity was detected in all Candida isolates. Only 21.6% of C. albicans from patients with diabetes had esterase activity as + 3, while it ranged from + 1 to + 2 in others. Hemolytic activity was determined in C. albicans, C. dubliniensis, C. glabrata and C. krusei as 0.79, 0.58, 0.66 and 0.74, respectively. Hemolytic activity was significantly different in the two groups of diabetics and non-diabetics. Oral carriage of C. albicans in the diabetic group (n = 42; 66.7%) was significantly greater than the control group (n = 16; 57.1%). Esterase activity of C. albicans in diabetic group was higher than non-diabetic group. Although C. albicans remains the most frequently pathogenic yeast for human, but other species are increasing.

Candiduria is a rising condition among hospitalized patients and Candida albicans is the most common recovered agent. However, non-albicans Candida species (NACs) such as C. glabrata, C. krusei, C. parapsilosis, and C. tropicalis are also important. Although most Candida species especially C. albicans are sensitive to routinely used antifungals, an increasing trend in resistance has been observed among NACs. The aim of the present study was to detect the susceptibility of Candida strains recovered from candiduria in hospitalized patients against posaconazole and caspofungin. A total of 120 urine samples were taken from patients hospitalized in Intensive Care Units (ICUs) (65) and urology (55) wards. All recovered yeasts were differentiated by using CHROMagar Candida medium and routine tests for identification of Candida species. Minimal inhibitory concentrations (MICs) of all isolates towards posaconazole and caspofungin were determined using the microdilution method with serial dilutions from 8 to 0.0625 µg/mL (posaconazole) and 4 to 0.03125 µg/ mL (caspofungin). In total, 41.7% of urine samples were positive for Candida isolation, including C. albicans (46%), C. glabrata (24%), C. tropicalis (16%) and C. krusei (14%). The MIC of caspofungin for 90% of the tested isolates was lower than 2 µg/mL. Furthermore, 94% of the tested isolates were inhibited by posaconazole at lower than 2 µg/mL after 24 hours, whereas 6% of isolates had MICs of more than 4 µg/mL. This study demonstrates the importance of Candida species in urine samples from hospitalized patients in ICUs and urology wards. It showed that both tested antifungals had excellent effects on different species of Candida, however the strains from ICUs were found to be more sensitive to caspofungin than posaconazole.

Helicobacter pylori are becoming increasingly recognized as a possible pathological cause of chronic rhinosinusitis (CRS). Considering the prevalence of CRS and its impact on quality of life, we decided to determine the role of H. pylori in chronic sinus infections by using the PCR technique.Patients and In a case-control analytical epidemiologic survey, the study population was selected by consecutive sampling from patients with CRS undergoing endoscopic sinus surgery during years 2010 - 2012. Patients were divided into two groups. The study group consisted of patients with CRS and the control group consisted of patients with nasal obstruction caused by concha bullosa, without inflammation or infection of the sinuses. Sampling was performed during surgery from the infected tissue and from the middle turbinate mucosa. Eventually, bacterial DNA was extracted and used for the PCR test, in order to isolate H. pylori. Nine patients (18%) with CRS had H. pylori isolated from their samples whereas in the control group, H. pylori were only found in two patients (4%); this difference was statistically significant (P = 0.025). The indicator wasn’t statistically significant between males and females. There was no statistical correlation in relative frequency of H. pylori for different age groups (P > 0.05). There was a significant correlation between CRS and presence of H. pylori in sinonasal mucosa. This relationship may reflect the role of H. pylori as one of the pathogenic factors in the development of CRS. However, further studies are required to confirm this role.

Chronic hepatitis B consists of different clinical phases. Laboratory and histological assessments can help differentiate the clinical phases of this disease and thus lead to better management. This study was conducted to determine laboratory and histological characteristics of HBeAg-negative and HBeAg-positive chronic hepatitis B patients.Patients and In this study, we evaluated 151 treatment naive chronic hepatitis B patients and grouped them according to their HBeAg status. Serum hepatitis B virus (HBV) DNA and HBsAg levels were measured, and liver function tests, and liver biopsy were performed for the study population. There was a significant difference in age, and HBV DNA and HBsAg levels between HBeAg-negative and HBeAg-positive groups yet there was no statistically significant difference in sex, liver function tests, grading and staging of liver biopsy between the groups. Hepatitis B virus DNA and HBsAg levels were correlated in both HBeAg-negative and HBeAg-positive chronic hepatitis B patients. We concluded that chronic hepatitis B patients had different HBV DNA and HBsAg levels according to their HBeAg status.