Inter-and intraspecific variations of leishmania strains isolated from patients with cutaneous and visceral leishmaniases in pars province, south of Iran

Background: Cutaneous and visceral leishmaniases are present in Fars Province in the south of Iran. The current study aimed to evaluate the inter- and intragenic diversities of Leishmania species isolated from patients with leishmaniasis in Fars Province, using PCR-based analyses and DNA sequencing of the N-acetylglucosamine-1-phosphate transferase [nagf] gene

Methods: Clinical samples were taken from the skin lesions of 120 individuals with clinical suspicion of cutaneous leishmaniasis [CL] referred to the major health centers of Shiraz. Along with microscopic examination, a part of each sample was used for in vitro cultivation. DNA was extracted from the cultured parasites and the nagt gene was PCR-amplified. For RFLP analysis, the PCR product of the nagt gene was digested with the Accl restriction enzyme. Moreover, the PCR products of 23 isolates were sequenced and analyzed, using MEGA5

Results: From the 120 patients with clinical suspicion of CL, 110[91.7%] cases were found to be positive by direct microscopy while 77[64.1%] of the cultures were positive. Digestion of the PCR product with the Accl restriction enzyme detected L. major in 57 out of the 77 [74.1%] and Z. tropica, in 20 out of the 77 [25.9%] cases with CL. Phylogenetic analysis grouped the Leishmania isolates into 3 main clades, representing L. major, L. infantum, and L. rop/ca,encompassing 2, 2, and 2 haplotypes, respectively. Within the clades, the L. tropica intraspecies divergence was more pronounced in L. major

Conclusion: The findings of this study demonstrated that the causative agent of CL in Fars Province was mainly L. major and that there was considerable heterogeneity between the Leishmania species and also within the L. major isolates

Knowledge, attitude, and practices related to cutaneous leishmaniasis in an endemic focus of cutaneous leishmaniasis, southern Iran

Objective: To assess knowledge, attitudes, and practices (KAP) of inhabitants of an endemic area in Fars province toward cutaneous leishmaniasis (CL). Methods: The study was carried out in Lapui district in Fars province, south of Iran, one of the most important foci of CL in this province. Sample size (237 residents) was calculated based on population. House-to-house survey was done to collect the data regarding knowledge, attitudes, and practices of the inhabitants. The head of each household was interviewed by a trained staff to assess his/her KAP related to CL. A semi-structured KAP questionnaire was used for data collection. Results: Mean age of participants was 39 and more than half of the respondents were in the age group of 31-40. Males constituted 172 (72.5%) of subjects. Most of the respondents (84.3%) were literate. The majority of the study population (83%) had heard about Salak (local name for CL) and most of these respondents (91%) were aware that CL is presented with a cutaneous lesion. Nearly two-third of the participants (63.5%) stated the bite of mosquito (not specifically sandflies) for CL transmission. The respondents' attitude regarding the treatment of CL was not satisfactory since only 48% believed that CL can be treated by medicine. A noticeable proportion of respondents (21%) believed in indigenous medicine for the treatment of CL. A small proportion of respondents (14%) stated that traditional healers are good at treating this disease. More than two-third (69%) of respondents believed that the disease is preventable although most of interviewees did not know about preventive measures. Conclusions: In this study, insufficient knowledge of community about infection nature, vector, transmission mode and preventive measures of CL, highlights the needs for a health education initiative to enhance the awareness of people about CL. This would improve inhabitants' contribution in control program of CL in this area.

Knowledge, attitude, and practices related to cutaneous leishmaniasis in an endemic focus of cutaneous leishmaniasis, Southern Iran

<p><b>OBJECTIVE</b>To assess knowledge, attitudes, and practices (KAP) of inhabitants of an endemic area in Fars province toward cutaneous leishmaniasis (CL).</p><p><b>METHODS</b>The study was carried out in Lapui district in Fars province, south of Iran, one of the most important foci of CL in this province. Sample size (237 residents) was calculated based on population. House-to-house survey was done to collect the data regarding knowledge, attitudes, and practices of the inhabitants. The head of each household was interviewed by a trained staff to assess his/her KAP related to CL. A semi-structured KAP questionnaire was used for data collection.</p><p><b>RESULTS</b>Mean age of participants was 39 and more than half of the respondents were in the age group of 31-40. Males constituted 172 (72.5%) of subjects. Most of the respondents (84.3%) were literate. The majority of the study population (83%) had heard about Salak (local name for CL) and most of these respondents (91%) were aware that CL is presented with a cutaneous lesion. Nearly two-third of the participants (63.5%) stated the bite of mosquito (not specifically sandflies) for CL transmission. The respondents' attitude regarding the treatment of CL was not satisfactory since only 48% believed that CL can be treated by medicine. A noticeable proportion of respondents (21%) believed in indigenous medicine for the treatment of CL. A small proportion of respondents (14%) stated that traditional healers are good at treating this disease. More than two-third (69%) of respondents believed that the disease is preventable although most of interviewees did not know about preventive measures.</p><p><b>CONCLUSIONS</b>In this study, insufficient knowledge of community about infection nature, vector, transmission mode and preventive measures of CL, highlights the needs for a health education initiative to enhance the awareness of people about CL. This would improve inhabitants' contribution in control program of CL in this area.</p>

Molecular genotyping of toxoplasma gondii in human spontaneous aborted fetuses in Shiraz, southern Iran

Congenital toxoplasmosis is associated with variable complications including encephalitis, microcephaly, hydrocephaly, hepatitis, lymphadenopathy and even intrauterine death. Presence of Toxoplasma gondii in human placenta may induce congenital infection. The aim of this study was to determine the genotypes of Toxoplasma gondii infection in human spontaneous aborted fetuses in Shiraz, south of Iran. Five hundred and forty two paraffin-embedded blocks of aborted placenta were collected, from two university-affiliated hospitals in Shiraz. Occurrence of spontaneous abortion was confirmed by examine of the slides. After re-cutting of the blocks and dewaxing, semi-nested PCR assay was used to detect the fragments of T. gondii B1 gene in the samples. Also direct molecular genotyping was performed on positive samples with Restriction Fragment Length Polymorphism-PCR analysis on the SAG2 gene. Among the 542 tissue samples, the B1 gene was amplified from 78 [14.4%] of cases with the semi nested PCR and typed by RFLP. The genotype of Toxoplasma strains of 65 [out of 78] PCR-positive samples were evaluated and 54 out of 65 [83.1%] were found to be type II and 11 out of 65 [16.9%] were type I. Considering the high level of Toxoplasma infection in aborted fetuses in this study, Toxoplasma might largely contribute to spontaneous abortion in this area of Iran

Nested polymerase chain reaction and sequencebased detection of leishmania infection of sand flies in recently emerged endemic focus of zoonotic cutaneous leishmaniasis,southern iran

Background: Geographical distribution of zoonotic cutaneous leishmaniasis [ZCL] has continuously been extended in recent years in Iran. The Beiza District is one of the newly-emerged endemic foci of ZCL in southern Iran. The main aim of the present study was to detect the vector[s] of ZCL in this area

Methods: To detect the fauna and vectors of ZCL in this district, sand flies were caught using sticky papers. Seventy randomly selected female sand flies out of 730 were molecularly investigated for Leishmania infection using species-specific nested polymerase chain reaction [PCR] assay between April and October 2010

Results: A total of 2543 sand flies were caught. The fauna was identified as 10 species [five Phlebotomus spp. and five Sergentomyia spp.]. Phlebotomus papatasi was the most dominant species both indoors and outdoors [37.55% and 16.35%, respectively]. L. major was detected in 5 out of 48 investigated Phlebotomus papatasi [10.41%]. Sequence-based characterization was carried out to confirm the PCR findings. The positive samples were shown to have 75-88% similarity with L. major sequences in GenBank

Conclusion: According to the findings of the present study, similar to the other foci of ZCL in Iran, P. papatasi is the proven and primary vector of CL. This study could be drawn upon for future strategy planning in this newly emerged endemic focus

High prevalence of hepatitis C infection among high risk groups in Kohgiloyeh and Boyerahmad province, southwest Iran

Background: Detection of Hepatitis C virus [HCV]-infected people in each community assists with infection prevention and control. This study aims to evaluate the prevalence of HCV infection among high risk groups in Kohgiloyeh and Boyerahmad Province, Southwest Iran

Methods: This was a cross-sectional study conducted from 2009-2010 in Kohgiloyeh and Boyerahmad Province. High risk groups for HCV were the subjects of this study. Blood samples were taken from 2009 individuals at high risk for HCV that included inmates, injecting drug users [IDUs], health care workers, patients on maintenance hemodialysis, hemophilic patients, and those with histories of blood transfusions. Patients were residents of Yasuj, Gachsaran, and Dehdasht [3 main townships in the province]. Samples were analyzed by ELISA for anti-HCV antibodies. Demographic features of participants were recorded by a questionnaire during sample collection. Data were analyzed by SPSS version 13 software

Results: Of 2009 subjects, HCV antibodies were detected in 172 [8.6%]. Rate of infection was higher in males [11.4%] compared to females [3.2%]. Rate of infection in inmates was 11.7% while this rate was 42.4% in IDUs, 4.2% in health care workers, and 6.1% in thalas-semic patients. Significant correlation was found between HCV infection, history of imprisonment, and thalassemia

Conclusion: Results of this study have provided epidemiologic features of HCV and its risk factors in Kohgiloyeh and Boyerahmad Prov-ince, Southwest Iran. This information may assist in preventing the spread of HCV infection in this and other similar settings in the region. The findings of this study may help in improving surveillance and infection control in the community through management and monitoring of infected individuals

Immune responses to antigens of in vitro reared echinococcus granulosus adult worms in Balb/c mice

Cystic echinococcosis [CE], also known as echinococcosis/hydatidosis, is one of the most important parasitic diseases in the world. It enhances both humoral and cellular [Thl and Th2] responses in infected host. Different antigens of the worm may favor the Thl or Th2 immune responses in CE patients. To evaluate the humoral and cellular immune responses of Balb/c mice against the crude and excretory/secretory [E/S] antigens of in vitro reared Echinococcus granulosus adult worms. A total of 20 Balb/c mice divided into 5 groups of 4 mice each. Three groups of mice [n=4] were immunized with crude, E/S and an immunodominant antigen of in vitro reared Echinococcus granulosus adult worms on day 1 and 28. The fourth and the fifth groups were negative control groups and received PBS plus adjuvant, or nothing, respectively. Two weeks after the second injection, the mice were killed and their blood was collected for determining antibody responses, and their spleens were employed for proliferation assay. Total IgG were measured by indirect ELISA. Spleen cells of immunized mice were cultivated and exposed to different antigens of adult worms including E/S and crude antigens. Level of IFN-y, IL-12, IL-4 and IL-10 were measured in the recovered cell culture supernatants by capture ELISA. Total IgG assay showed the highest level of antibody produced in mice immunized with crude antigens. Proliferation assay showed a statistically significant production of cytokines in the mice immunized with crude antigens [p<0.05]. The highest levels of IFN-gamma, IL12 and IL-4 were produced in mice immunized with crude antigen of the in vitro reared Echinococcus granulosus adult worms followed by E/S antigens. Immunodomonant antigen induced the lowest levels of cytokines [IL-12, IFN-gamma, IL-4 and IL-10] in immunized mice. A significant levels of Thl related cytokines [IFN-gamma and IL-12] were produced in Balb/c mice immunized with crude antigen of the in vitro reared Echinococcus granulosus adult worms

leishmania infantum fml-elisa for the detection of symptomatic and asymptomatic canine visceral leishmaniasis in an endemic area of Iran

Visceral leishmaniasis [VL] is caused by Leishmania infantum in Mediterranean basin and is an endemic disease in some parts of Iran. Canines are the main reservoirs of VL in most of the endemic areas. Different serological methods have been introduced for diagnosis of canine visceral leishmaniasis [CVL]. In this survey a Fucose-Mannose Ligand [FML] ELISA, using native L. infantum antigen, was developed and its validity for detection of infected dogs in comparison with direct agglutination test [DAT] and PCR was evaluated. Blood samples of sixty ownership dogs [/= 1/320] in DAT while seven of the 60 [11.66%] samples were positive by FML-ELISA. Nine out of 60 [15%] buffy coat samples showed a band about 680 bp indicative of L. infantum in PCR. Three out of 60 dogs had Kala-azar symptoms and were positive by PCR and FML-ELISA, while two of these three dogs had antibody titers >/= 1/320 in their serum samples. The sensitivity and specificity of FML-ELISA for the detection of CVL in both symptomatic and asymptomatic dogs were found to be 77.8% and 100%, respectively. Considering the acceptable sensitivity and high specificity of FML-ELISA, use of this serological method can be recommended for epidemiological surveys of CVL

Performance of latex agglutination test [KAtex] in diagnosis of visceral leishmaniasis in Iran

Visceral leishmaniasis [VL] is an endemic disease in some parts of Iran. Many techniques have been used for diagnosis of VL, among which the urine based latex agglutination test [KAtex] is a promising one. To compare three diagnostic tests of VL including KAtex, ELISA and Direct Agglutination Test [DAT] in VL patients and healthy controls in the south west of Iran. Serum [n = 29] and urine samples [n = 31] were collected from parasitologically confirmed VL patients. Control samples were obtained from healthy individuals [n = 61] and also from patients with infectious diseases other than VL. The collected serum samples were tested by DAT and ELISA using crude antigen from promastigotes of Leishmania infantum and the urine samples were tested by KAtex. Sensitivity and specificity of KAtex for diagnosis of VL was found to be 83.9% and 100%, respectively. Sensitivities of DAT and ELISA were 93.1% and 86.2% and their specificities were 100% and 90.5%, respectively. KAtex yielded a satisfactory sensitivity and specificity in diagnosis of VL in Iran and can be recommended as a rapid, field applicable and reliable test for diagnosis of VL in this region

Serum Antigen and Antibody Detection in Echinococcosis: Application in Serodiagnosis of Human Hydatidosis

Diagnosis of hydatidosis is based on immunodiagnostic methods along with radiological and ultrasound examinations. The objectives of the present study were to develop a specific and simple antigen-based ELISA method for diagnosis of hydatidosis and compare it with antibody detection method. The subjects in this study included 89 patients in the following groups: surgically confirmed hydatidosis patients (35 cases), control with other parasitic diseases (29 cases), and healthy controls (25 cases). Hyperimmune serum was raised against hydatid cyst fluid in rabbits. Anti-hydatid cyst IgG was purified by affinity chromatography using protein A column and labeled with horseradish peroxidase. Collected sera were assessed for hydatid cyst antigens and antibody by ELISA. Circulating hydatid antigen was found in 9 out of 35 patients with surgically confirmed hydatidosis. A sensitivity of 25.7% and a specificity of 98.0% were calculated for the antigen detection assay. Antibody detection by indirect ELISA, using antigen B, showed that 94.2% of patients (33 cases) have anti-hydatid cyst antibodies in their serum while cross reaction was noted in a few of non-hydatidosis patients. A sensitivity of 94.2% and specificity of 81.6% were found for the antibody detection assay. Findings of this study indicated that antibody detection assay is a sensitive approach for diagnosis of hydatid cyst while antigen detection assay might be a useful approach for assessment of the efficacy of treatment especially after removal of the cyst.

Comparison of serological methods [ELISA, DAT and IFA] for diagnosis of visceral leishmaniasis utilizing an endemic strain

The causative agent of visceral leishmaniasis [VL] in Iran is Leishmania infantum [L. infantum] [Mediterranean type] and its major reservoir host is the dog. To compare the serological methods including direct agglutination test [DAT], indirect immunofluorescent-antibody test [IFA] and enzyme-linked immunosorbent assay [ELISA] for serodiagnosis of endemic strain of L. infantum. 61 blood samples from VL patients referred to Shiraz hospitals and 49 blood samples from control group were collected. Native strain of the parasite isolated from a VL patient from the region was cultured and characterized. Antigens from this L. infantum parasite were used in ELISA and IFA system. Anti-Leishmania antibody was detected in 43 [70.5%], 49 [80.3%] and 51 [83.6%] cases using DAT, IFA and ELISA, respectively. Based on these results, sensitivity and specificity of DAT was found to be 70.5% and 100%, respectively. Sensitivities of IFA and ELISA in diagnosis of VL were 80.3% and 83.6% and their specificity was 90.5%. Results of this study showed that DAT and ELISA have the highest specificity and sensitivity in diagnosis of VL. DAT is a simple, cost-effective and field applicable test. Thus, it can be recommended for early and accurate diagnosis of VL, especially in regions where malaria, brucellosis and tuberculosis are prevalent

Comparative efficacy of dexamethasone versus hydrocortisone in severe acute pediatric asthma

This study aimed to compare the efficacy of dexamethasone and hydrocortisone in the treatment of acute paediatric asthma. Forty seven asthmatic children were randomised to receive either dexamethasone [0.6 mg/ml, maximum 16 mg] in a single dose or hydrocortisone [8-10 mg/kg/day] in 4 equal doses for 2 days. Our findings showed that the mean length of hospitalization in children receiving dexamethasone was significantly shorter than those receiving hydrocortisone

Prevalence of Toxocara cati and other intestinal helminths in stray cats in Shiraz, Iran

Toxocariasis is a parasitic zoonosis with worldwide distribution that affects both cats and dogs. Necropsy of 114 stray cats from Shiraz revealed that 106 (92.9%) stray cats were infected at least with one of the intestinal helminth species. The overall infection rates in stray cats infected with cestoda and nematoda were 105(99.1%) and 101(95.3%) respectively. The detected cestodes were Joyeuxiella pasqualei (34.3%), Dipylidium caninum (49.5%), Taenia taeniaeformis (12.3%), Spirometra sp. (3.8%) and the detected nematodes were Physaloptera sp. (44.6%), Toxocara cati (42.6%) and Toxascaris leonina (12.9%). The study revealed that T. cati was one of the most frequently detected intestinal helminths, which is an important source of zoonotic helminths.