The Occurrence of the Strongylid Nematodes, Kalicephalus viparae viparae (Nematoda: Diaphanocephalidae), in Viper Snakes, Macrovipera lebetina (Reptilia: Viperidae), Southwestern Iran.

ancylostomatid Kalicephalus spp. is the common parasitic intestinal nematode of reptiles. West-Asian blunt-nosed viper is a venomous snake found in extensive areas of Iran. From June to September 2017, two dead viper snakes were referred to a parasitology laboratory and examined for intestinal parasites. Several white elongated roundworms were collected and fixed to identify under light and scanning electron microscopes (SEM) based on morphological and molecular characteristics. For the molecular survey, some parts of the identified worms were extracted and the ITS of nuclear ribosomal DNA (rDNA) was amplified by polymerase chain reaction (PCR). Five roundworms were found in one snake and three worms with similar morphological characteristics in another one. All the collected female hookworms were taxonomically identified as Kalicephalus viperae viperae. The SEM findings showed the head was small and had three dorsal, ventral, and middle circumoral papillae with a spike-like process on the median papilla of K. viperae. Moreover, the buccal capsule was bivalvular and included two lateral valves consisting of several chitonid pieces. The tail of the female worm was slim and long with a blunt end and had a terminal spike at its end. In the molecular survey, the ITS of rDNA amplified at about 850 bp was identified as K. viperae. The ITS gene rDNA phylogeny analysis of the K. viperae sequence showed that the isolated species had high similarity to Ancylostoma species from around the world and is close to Ancylostoma braziliense with 88% discrepancies in the phylogenetic tree. The morphological characteristics and a large part of K. viperea viperea rDNA nucleotide sequence were reported in viper snakes for the first time in the world and in Iran.

A Preliminary Study on the Antigenic Proteins of Linguatula serrata Nymphal Stage in Sheep with Visceral Linguatulosis.

Scant information is available on the immunological aspect of Linguatula serrata causing linguatulosis in humans and animals. The present study aimed to analyze the content of crude somatic extracts and excretory-secretory products of L. serrata nymphs to detect the immune response of sheep and immunogenic proteins of the parasite. After collecting the nymphs, somatic extracts were prepared by sonication. Excretory secretory products were prepared by the incubation of nymphs in RPMI medium at 37°C with 5% CO2. Somatic and excretory-secretory proteins were isolated using SDS-PAGE. The immunogenic properties of the resulting proteins were determined using immunoblotting and positive sera from sheep infected with visceral linguatulosis. The total content of somatic extracts and excretory-secretory products of L. serrata nymphs analyzed by SDS-PAGE (12% gel) revealed two protein patterns with more than 18 and 9 strong bands, respectively. Immunoblots using sera samples of sheep infected with the parasite, somatic extracts and excretory-secretory products demonstrated 12 and 3 antigenic proteins with molecular weights mostly in the range of 24-100 kDa and an antigen more than 180 kDa. Three common immunodominant antigenic proteins with molecular weights of 38 and 57, as well as an antigen of more than 180 kDa, were detected in the somatic extracts and excretory-secretory products of L. serrata nymphs in sheep with visceral linguatulosis. These antigens can be considetered prime candidates for future serodiagnosis and immunoprotective studies of the parasite.

Orbital mucormycosis in an immunocompetent individual.

BACKGROUND: Orbital mucormycosis caused by Zygomycetes is a rare and fatal infection that generally affects the patients who are immunocompromised. Despite antifungal therapy and aggressive surgical intervention, mucormycosis can cause serious and rapidly fatal infections if delayed diagnosis or therapeutic management occurs. Here, we report orbital mucormycosis in a healthy boy, with a favorable outcome after aggressive treatment. He has had no recurrence since the end of his treatment. CASE PRESENT: A 2-year old healthy boy, some days after entry of dust particle to his left eye presented with swelling and redness of the eye. With diagnosis of "periorbital cellulitis" intravenous antibiotics vancomycin (40 mg/kg/day) and ceftriaxone (75 mg/kg/day) were started but no improvement was observed. The results of biopsy and tissue culture led us to a diagnosis of mucormycosis. Orbital exenteration, combined with intravenous amphotericin B (1 mg/kg/day), resulted in the patient's survival. CONCLUSION: Due to the high mortality rate of mucormycosis, early diagnosis based on clinical findings and biopsy could be effective for management of the patients suffering from this infection.

Genotypic Analysis of E. coli Strains Isolated from Patients with Cystitis and Pyelonephritis.

BACKGROUND: Urinary tract infection is the most common health problem affecting millions of people each year, mainly caused by a large genetically heterogeneous group of Escherichia coli called uropathogenic E. coli This study investigates the genotypic analysis of E. coli strains isolated from patients with cystitis and pyelonephritis. METHODS: During 2008-2009, 90 E. coli strains were analyzed, consisting of 48 isolates causing pyelonephritis in children and 42 isolates causing cystitis. Having identified the strains by standard methods, they were subtyped by pulsed field gel electrophoresis (PFGE) and their corresponding patterns were compared using dendrogram. RESULTS: Sixty five PFGE profiles were obtained from the genome of E. coli strains by this genotyping method. Thirty six and thirty three patterns were obtained for pyelonephritis and cystitis, respectively. Most strains exhi-bited twelve and thirteen bands and the patterns with eight or nineteen bands had the lowest rate. Genome sizes of the strains were between 1610-4170 kbp. CONCLUSION: With due attention to these results, genetic patterns showed that the strains had different clonalities and it could be suggested in some cases that the strains causing pyelonephritis or cystitis have common patterns and different diseases could be explained by different gene factors.

Vancomycin-Resistant Entrococci colonization in chronic hemodialysis patients and its risk factors in southern Iran (2005-2006).

BACKGROUND: Vancomycin-resistant enterococci (VRE) recently have emerged as a nosocomial pathogen among dialysis patients. This study aims to appraise the prevalence, incidence density and risk factors for VRE colonization among these patients. METHODS: In this prospective study, 782 stool or rectal swab specimens were collected from 250 chronic hemodialysis patients with an interval of at least one month. To identify the risk factors of VRE colonization, demographic and health data of VRE+ and VRE- patients were compared. RESULTS: VRE colonization was detected in 55 (22%) patients during study. Incidence density of one case per 79.6 patient-month of follow up was estimated.The only significant difference between the data collected from VRE+ and VRE- patients was observed in antibiotic consumption (P<0.001). CONCLUSION: VRE colonization is relatively high and rapidly spreading among chronic dialysis patients. It is strongly associated with recently antibiotic consumption.

Susceptibility of Candida species isolated from immunocompromised patients to antifungal agents.

This study was carried out from October 2003 to March 2007 to investigated susceptibility patterns ot March 2007 to antifungals of Candida strains isolated from 410 immunocompromised patients in Shiraz, Islamic Republic of Iran. Patients were checked for systemic candidiasis. Fungal colonization was determined and clinical samples collected from those patients with clinical signs of infections were examined. The carbohydrate assimilation patterns of all 354 isolates were studied. Susceptibility of the isolates to antifungal agents was determined using the reference broth microdilution method. Candida albicans was the species most often isolated. Voriconazole was highly active against all the isolates. Major resistance to itraconazole was observed in all Candida spp. Regular investigations into antifungal resistance in medical centres is highly recommended as this will result in more efficient management of invasive candidiasis in immunocompromised patients.

Evaluation of cytomegalovirus infection after six months of liver transplantation in children in shiraz, southern iran.

BACKGROUND: Liver transplantation (LT) is a life-saving treatment for end-stage liver diseases (ESLD). Cytomegalovirus (CMV) infection is one of the important causes of morbidity after LT. OBJECTIVE: To evaluate the incidence of late-onset (after 6 months of LT) CMV infection in pediatric recipients. METHODS: A retrospective analysis was conducted to evaluate 50 pediatric patients who underwent LT for 8 years at the LT Unit of Nemazee Hospital affiliated to Shiraz University of Medical Sciences, Shiraz, Iran. We retrospectively investigated episodes of CMV infection after 6 months of LT proven by CMV antigenemia test. RESULTS: Three recipients (6%) developed late-onset CMV infection. These patients finally responded to ganciclovir. CONCLUSION: CMV infection is one of the most common post-LT viral infections that usually occurs in the first six months of LT. Our study shows that the incidence of late-onset CMV infection is relatively low, but it still remains a significant problem. Therefore, monitoring and management is crucial for improving the survival of children.

Susceptibility of clinical Candida species isolates to antifungal agents by E-test, Southern Iran: A five year study.

BACKGROUND AND OBJECTIVES: The incidence of fungal infections in immunocompromised patients, especially by Candida species, has increased in recent years. This study was designed to identify Candida species and determine antifungal susceptibility patterns of 595 yeast strains isolated from various clinical specimens. MATERIAL AND METHODS: Identification of the isolates were determined by the API 20 C AUX kit and antifungal susceptibilities of the species to fluconazole, amphotericin B, ketoconazole, itraconazole, voriconazole, and caspofungin were determined by the agar-based E-test method. RESULTS: Candida albicans (48%) was the most frequently isolated species, followed by Candida kruzei (16.1%), Candida glabrata (13.5%), Candida kefyr (7.4%), Candida parapsilosis (4.8%), Candida tropicalis (1.7%) and other species (8.5%). Resistance varies depending on the species and the respective antifungal agents. Comparing the MIC90 for all the strains, the lower MIC90 was observed for caspofungin (0.5 µg/ml). The MIC90 for all Candida species were 64 µg/ml for fluconazole, 0.75 µg/ml for amphotericin B, 4 µg/ml for ketoconazole, 4 µg/ml for itraconazole, and 2 µg/ml for voriconazole. CONCLUSIONS: Species definition and determination of antifungal susceptibility patterns are advised for the proper management and treatment of patients at risk for systemic candidiasis. Resistance to antifungal agents is an alarming sign for the emerging common nosocomial fungal infections.

Susceptibility of Candida species isolated from immunocompromised patients to antifungal agents

This study was carried out from October 2003 to March 2007 to investigate susceptibility patterns to antifungals of Candida strains isolated from 410 immunocompromised patients in Shiraz, Islamic Republic of Iran. Patients were checked for systemic candidiasis. Fungal colonization was determined and clinical samples collected from those patients with clinical signs of infections were examined. The carbohydrate assimilation patterns of all 354 isolates were studied. Susceptibility of the isolates to antifungal agents was determined using the reference broth microdilution method. Candida Candida albicans was the species most often isolated. Voriconazole was highly active against all the isolates. Major resistance to itraconazole was observed in all Candida spp. Regular investigations into antifungal resistance in medical centres is highly recommended as this will result in more efficient management of invasive candidiasis in immunocompromised patients

Prevalence of oropharyngeal colonization by Haemophilus influenzae type b in Iranian children.

Healthy carriers of Haemophilus influenzae type b (Hib) play an important role in the spread of invasive disease. The aim of this study was to assess the need for Hib vaccination in Iranian children by estimating the prevalence of Hib oropharyngeal colonization among children in Tehran. Cultures were prepared from oropharyngeal swabs of 1000 children in 25 day-care centres in Tehran from October 2005 to March 2006. The prevalence of Hib carriers was 7.6%, similar to other developing countries prior to inoculation with the conjugate Hib vaccine. We recommend Hib vaccination be included in the Iranian national programme of immunization.

Increased levels of IL-10, IL-12, and IFN- in patients with visceral leishmaniasis.

Visceral leishmaniasis remains a serious public health problem in developing countries. Cytokines have a crucial role in the pathogenesis of this disease. We evaluated plasma levels of IL-10, IL-12 and IFN- in 32 patients with active visceral lieshmaniasis, in 29 siblings of the patients and in 23 normal individuals in an endemic area to look for correlations between the clinical outcome of infection and the plasma cytokine levels. An analysis was made with a skin test and a quantitative sandwich enzyme immunoassay. Data were analyzed with the Mann Whitney test and the Kruskal Wallis test. The cytokine levels were significantly higher in the patients than in the control groups. We concluded that normalization of the plasma level of IFN- can serve as a reliable parameter for considering the patient as cured.

Increased levels of IL-10, IL-12, and IFN- in patients with visceral leishmaniasis

Visceral leishmaniasis remains a serious public health problem in developing countries. Cytokines have a crucial role in the pathogenesis of this disease. We evaluated plasma levels of IL-10, IL-12 and IFN- in 32 patients with active visceral lieshmaniasis, in 29 siblings of the patients and in 23 normal individuals in an endemic area to look for correlations between the clinical outcome of infection and the plasma cytokine levels. An analysis was made with a skin test and a quantitative sandwich enzyme immunoassay. Data were analyzed with the Mann Whitney test and the Kruskal Wallis test. The cytokine levels were significantly higher in the patients than in the control groups. We concluded that normalization of the plasma level of IFN- can serve as a reliable parameter for considering the patient as cured.

Prevalence of oropharyngeal colonization by Haemophilus influenzae type b in Iranian children

Healthy carriers of Haemophilus influenzae type b [Hib] play an important role in the spread of invasive disease. The aim of this study was to assess the need for Hib vaccination in Iranian children by estimating the prevalence of Hib oropharyngeal colonization among children in Tehran. Cultures were prepared from oropharyngeal swabs of 1000 children in 25 day-care centres in Tehran from October 2005 to March 2006. The prevalence of Hib carriers was 7.6%, similar to other developing countries prior to inoculation with the conjugate Hib vaccine. We recommend Hib vaccination be included in the Iranian national programme of immunization

Molecular detection of invasive aspergillosis in hematologic malignancies.

BACKGROUND: Aspergillus species are the most frequent causes of invasive mold infections in immunocompromised patients, particularly those who underwent chemotherapy for hematologic malignancies. The aim of this study was to determine the incidence and efficiency of the PCR-enzyme linked immunosorbent assay method (PCR-ELISA) for early detection of Aspergillus species in patients with hematologic malignancies. PATIENTS AND METHODS: From 2004 to 2006, 194 patients with hematologic malignancies (who received chemotherapy) were evaluated for invasive aspergillosis (IA) in Shiraz, southern Iran. Ethylenediaminetetraacetic acid anticoagulant whole blood samples were collected prospectively once a week and stored at -20 degrees C until examination. All collected blood samples were assayed for the presence of the bands on ethidium bromide stained gel and for hybridization. RESULTS: The female-to-male ratio was 61:133, the mean age of patients was 33.7 years, and mean of hospitalization period was 21.2 days. PCR-ELISA was positive in 14 (7.2%) patients who exhibited clinical and radiologic signs of IA. The etiologic agents were Aspergillus flavus (11 cases) and Aspergillus fumigatus (three cases). The mean time of positivity of PCR-ELISA in the blood before the appearance of clinical signs was 12.6 days. PCR was found to be the earliest indicator of IA preceding nonspecific clinical and radiologic findings. The sensitivity, specificity, positive, and negative predictive values of PCR-ELISA to detect DNA-specific for Aspergillus species in patients with proven and probable IA were 66%, 96%, 62.5%, and 97%, respectively. In case patients were treated with antifungal drugs, and the treatment was successful, fungal PCR assay became negative after 14 days and if the treatment failed, assay was positive until death. CONCLUSIONS: We demonstrated, in the present study, the incidence of IA in leukemic patients and the usefulness of molecular assay for early diagnosis and monitoring of the treatment of IA.

Meningococcal carrier rate before and after hajj pilgrimage: effect of single dose ciprofloxacin on carriage.

We determined the carriage rate of Neisseria meningitidis before and after hajj pilgrimage among a group (1) of 674 randomly selected Iranian pilgrims, and the effect of 500 mg of ciprofloxacin given 24 hours before return on the reduction of meningococcal carriers among another group (2) of 123 randomly selected Iranian pilgrims. Throat specimens taken 1 hour before departure on the hajj and immediately on return were cultured. Carriage rates of N. meningitidis in group 1 were 5.2% before and 4.6% after pilgrimage (P = 0.65); 3 new serogroups (Z, Z and A) were identified on return. In group 2, the carriage rate decreased from 8.1% to zero before and after pilgrimage.

Meningococcal carrier rate before and after hajj pilgrimage: effect of single dose ciprofloxacin on carriage

We determined the carriage rate of Neisseria meningitidis before and after hajj pilgrimage among a group [1] of 674 randomly selected Iranian pilgrims, and the effect of 500 mg of ciprofloxacin given 24 hours before return on the reduction of meningococcal carriers among another group [2] of 123 randomly selected Iranian pilgrims. Throat specimens taken 1 hour before departure on the hajj and immediately on return were cultured. Carriage rates of N. meningitidis in group 1 were 5.2% before and 4.6% after pilgrimage [P = 0.65]; 3 new serogroups [Z, Z' and A] were identified on return. In group 2, the carriage rate decreased from 8.1% to zero before and after pilgrimage

Evaluation of rK39 strip test for the diagnosis of visceral leishmaniasis in infants.

This study estimated the sensitivity and specificity of the rK39 strip test compared with the immunofluorescent antibody test and microscopy of bone marrow aspirate smears (the gold standard) in 47 children with suspected visceral leishmaniasis. A control group of children with other diagnoses (tuberculosis, toxoplasmosis, systemic lupus erythematosus, malaria or cutaneous leishmaniasis) were also tested to check false positive results. The sensitivity and specificity of the strip test were 82.4% and 100% and that of immunofluorescent antibody were 100% and 92.7%. The rK39 strip test is reliable where there is no access to laboratory facilities.

Improvement in isolation of human peripheral blood leukocyte subpopulations: application in diagnosing human cytomegalovirus infection in bone marrow transplant patients.

OBJECTIVES: High-yield isolation and purification of human leukocyte subpopulations from whole blood is fundamental to many biological and medical applications including qualitative and quantitative PCR-based techniques of determining human cytomegalovirus infection. Several procedures have been reported to purify morphologically and functionally intact human leukocyte subpopulations for diagnostic proposes. Here, we report and evaluate a technique for high-yield purification of intact and viable human leukocyte subpopulations based on modification of a previous methodology. MATERIALS AND METHODS: One hundred peripheral blood samples were collected from bone marrow transplant recipients (n = 60), bone marrow donors (n = 20), and healthy blood donors (n = 20). The samples were tested in parallel using 4 different leukocyte separation methods. The methods were evaluated based on the concentration, purity, and viability of the isolated leukocyte subpopulations. RESULTS: When compared with standard methods, our methods produced 99% purity for both polymorphonuclear or mononuclear leukocytes. The corresponding viability for the methods was determined to be 98%. No erythrocyte contamination was demonstrated. However, the maximum concentration for polymorphonuclear or mononuclear leukocytes obtained by standard methods was 70%. The corresponding viability for all the methods was determined to be 98%. CONCLUSIONS: Our results indicate that in patients with decreased whole blood leukocyte numbers, using either a modified Ficoll NH(4)Cl or a modified dextran method would be valuable for simultaneous separation of polymorphonuclear and mononuclear leukocytes with high purity, viability, and concentration.

Relation of Chlamydia pneumoniae infection to documented coronary artery disease in Shiraz, Southern Iran.

The possibility that infectious agents may trigger a cascade of reactions leading to inflammation, atherogenesis, and vascular thrombotic events has recently been raised. Chlamydia pneumoniae is one of those that have received the most investigative attention with respect to coronary artery disease (CAD). This study was undertaken for the first time in Shiraz, Iran to determine this relationship. A case-control study was conducted in 167 subjects (81 women and 86 men) who underwent coronary angiography at cardiac catheterization laboratories of Shiraz University of Medical Sciences Hospitals. Immunoglobulin G (IgG), and IgA antibodies to C. pneumoniae antigen were estimated in baseline serum samples from 109 patients (mean age 57 years) experiencing a coronary event and from their matched controls (n = 58, mean age 50 years) by ELISA method. The prevalence of IgG and IgA antibodies to C. pneumoniae did not show any case-control differences (82.6% vs 74.1% and 22% vs 15.5%, respectively). These results suggest that although C. pneumoniae was highly prevalent among these patients, it did not appear to be associated with angiographically documented CAD and cannot be regarded as a positive predictor for the development of acute coronary syndrome.

Comparative analysis of a double primer PCR assay with plasma, leukocytes and antigenemia for diagnosis of active human cytomegalovirus infection in bone marrow transplant patients.

The aim of the study was to determine the prognostic value of a double primer PCR assay to detect human cytomegalovirus (HCMV) infection or disease in bone marrow transplant (BMT) recipients. A total of 209 blood samples including peripheral blood mononuclear cells (PBMN), polymorphonuclear (PMN) leukocytes and plasma from 26 BMT recipients were tested by PCR assay. To discriminate between latent and active HCMV infection, 177 blood samples were also tested by a quantitative antigenemia assay. HCMV serology status of donors and recipients was determined before transplantation by an enzyme immunosorbent assay method. Using the double primer PCR assay, the number of positive samples increased by an average of 11.6%. Symptomatic active HCMV infection was diagnosed in 14 (53.8%) out of 26 BMT patients. There was a good association between double primer PCR assay of PMN leukocytes and antigenemia assays for detection of active HCMV infection in all patients. Detection of HCMV DNA in PMN leukocytes of BMT patients by double primer PCR assay can be an alternative method for antigenemia assay. However, quantitative PCR methods will be necessary for monitoring antiviral treatment.