Causes of Pediatric Visceral Leishmaniasis in Southeastern Iran

Leishmania infantum is the most frequent cause of visceral leishmaniasis and L. tropica has been rarely linked to the disease in Iran. In this study, bone marrow aspirates were collected from 10 child patients, suspected with visceral leishmaniasis referred to the Pediatric Wards of Kerman Medical Hospitals, Kerman, Iran during 2002–2011. Leishmania species were identified by using nested PCR in all slides. The PCR samples from nine patients indicated L. infantum as principal causative agent of visceral leishmaniasis and one L.tropica as a minor species.     

Genetic Diversity of Human Blastocystis Isolates in Khorramabad,Central Iran

Background

There are some genetic differences in Blastocystis that show the existence of species or genotypes. One of these genes that help in identifying Blastocystis is SSUrRNA. The aim of this study was assessment of genetic diversity of Blastocystis by PCR with seven pairs of STS primers.

Methods

This study was done on 511 stool samples collected from patients referred to the health care centers of Khorramabad, Central Iran, in 2012. Genomic DNA was extracted and in order to determine the Blastocystis subtype in contaminated samples, seven pairs of primers STS (subtype specific sequence-tagged site) were used.

Results

Out of 511 samples, 33 (6.5%) samples were infected with Blastocystis. Subtype (ST) of 30 samples was identified and three subtypes 2, 3 and 4 were determined. Mix infection was reported 10% which 3.33% of the infection was for the mixture of ST 3 and ST5 and 6.67% was for the mixture of ST 2 and ST 3.

Conclusion

The predominant subtype was ST3 that is the main human subtype. The dominance of ST2 and 5 are important in this study. This superiority has been reported in some of the studies in ST 2 which is different from the studies in other countries, because they have announced priorities of the ST1 and ST6 after ST3.     

Prevalence of Toxocara Spp. eggs in Public Parks in Tehran City,Iran

Background

The objective of the present research was to determine the frequency of Toxocara spp. eggs in soil samples of public parks, in the city of Tehran, Iran.

Methods

A total of 600 soil samples were taken from 120 parks between Aprils to November, 2008. Soil samples were collected from 5 distinct sites in the parks. The samples were washed with saline solution and the collected sediment from each park were equally divided and examined by floatation and Petri dish methods for Toxocara eggs.

Results

Ten percent were contaminated with Toxocara spp. eggs. The number of observed Toxocara eggs in each microscopic field was varied from 1-3. No significant differences were observed between floatation and Petri dish methods.

Conclusion

Our public parks showed a high risk of toxocariasis and the need for preventive studies.     

Visceral leishmaniasis in Iran: Review of the Epidemiological and Clinical Features

Visceral leishmaniasis (VL) is a life-threatening vector-borne parasitic disease is distributed in some parts of the new world and old world. The disease is endemic in different parts of Iran. This review article has been focused on major topics of epidemiological aspects and clinical features of VL in Iran for the period of 2002 through 2012. For the detection of VL in humans as well as animal reservoir hosts, anti-Leishmania antibodies were detected using direct agglutination test (DAT) as a validated serological test. Parasitological examinations were performed on suspected VL patients as well as canines and rodents. Different molecular methods were used for identification of species and genotype/ or strain of Leishmania spp. isolated from infected humans, animal reservoir hosts and vectors. Altogether, 1698 out of 36081 (4.7%) human serum samples collected from 5 distinct geographical zones showed anti-Leishmania antibodies at titers ≥ 1:3200 using DAT. The majority of VL cases in the endemic areas were found among children up to 12 years old. Almost 75% of DAT-positive cases (≥1:3200) in endemic areas showed clinical signs and symptoms. Predominant signs and symptoms in 217 hospitalized patients with DAT positive (≥1:3200) results included paleness (99.5%), fever (96.9%), splenomegaly (91.5%), hepatomegaly (53.6%) and lymphadenopathy (21.1%). Integrated VL surveillance system in primary care using DAT, could decrease mortality and morbidity of the disease in the VL endemic areas of the northwestern Iran. Out of 7204 serum samples collected from domestic dogs in various geographical locations of Iran, 879 (12.2%) were DAT sero-positive at titers ≥ 1:320. L. infantum as the principal causative agent of the disease was isolated from infected humans, domestic and wild canines and rodents. The principal animal reservoir hosts of the infection are domestic and wild canines. Ph. kandelakii, Ph. perfiliewi transcaucasicus, Ph. tobbi in northwestern Iran; Ph. major s.l. (=Ph. neglectus), Ph. keshishiani, and Ph. alexandri in southern parts of Iran were molecularly and/or parasitologically positive for L. infantum infections. The zoonotic form of VL (ZVL) caused by L. infantum occurs sporadically in all geographical zones of Iran but in northwestern and southern parts of the country the disease is endemic. DAT as an appropriate and potential tool has been used for sero-diagnosis and sero-epidemiological of VL among humans as well as domestic and wild canines.     

Molecular and Seroepidemiological Survey of Visceral Leishmaniasis among Humans and Domestic Dogs in Mazandaran Province,North of Iran

Background

New cases of visceral leishmaniasis (VL) have been reported recently in some parts of Mazandaran Province, north of Iran where the first human case of VL was reported in 1949. This study aimed to determine the present status of Leishmania infantum infection among humans and domestic dogs using serological and molecular methods in central parts of Mazandaran Province.

Methods

In this cross-sectional study, blood samples were randomly collected from 402 humans and forty-nine domestic dogs throughout 2009 and 2010 in the central part of Mazandaran Province including Semeskadeh and Kiakola districts where recent cases of human visceral leishmaniasis had been reported there. All the collected samples were tested by direct agglutination test (DAT) for the detection of anti-Leishmania infantum antibodies as well as convenience PCR assay on whole blood samples for detection of leishmanial infection and identification of Leishmania species.

Results

None of 402 collected human (402) and dog (49) blood samples showed anti Leishmania infantum antibodies at titers 1:3200 and 1:320 as cut-off values of DAT, respectively but only 2 of domestic dogs (4.1%) were found PCR-positive corresponding to L.infantum.

Conclusion

This study confirms the circulation of L. infantum at least among domestic dogs and highlights the sporadic pattern of VL in the studied areas. Further investigations regarding to sand flies fauna and wild canines as reservoir hosts of the disease, are recommended.     

Seroepidemiological Survey of Human Visceral Leishmaniasis in Ilam Province,West of Iran In 2013

Background:

Visceral leishmaniasis (VL) as one of the most important human parasitic disease is endemic in some parts of Iran. Several cases of VL have been reported recently in the Ilam Province. The current study aimed to assess the present status of human VL in the region.

Methods:

A random cluster sampling method was used to collect 456 serums samples from the children up to 12 years of age and 10% of adults living in urban and rural areas of the province. All the collected serum samples were tested by direct agglutination test (DAT) to detect anti- Leishmania infantum antibodies.

Results:

Of the examined 456 serum samples with direct agglutination test (DAT), only 21 (0.43%) sera showed anti- Leishmania antibodies at titers 1:400 and higher. Distribution of anti- Leishmania antibodies titers were: 1:400(n=4), 1:800(n=11), 1:1600(n=3), 1:3200(n=1), and 1:6400(n=1). Individuals with titers ≥1:3200 showed clinical signs and symptoms such as fever and splenomegaly. The highest and lowest seropositivity were observed in the age groups of 5–9 and >15 years old, respectively. There were no significant difference between the rate of seropositivity in males and females.

Conclusion:

VL with a low prevalence circulates in some parts of Ilam province, particularly in the southern parts. Complementary studies should be needed to find animal reservoir hosts and vectors. Furthermore, health systems and physicians should pay particular attention to the disease.     

Seroprevalence of Toxoplasma gondii Infection in Dogs in Tehran,Iran

Background

Toxoplasma gondii infects a wide range of animals; felines are definitive hosts and other animals including the dogs are intermediate hosts. The aim of this study was to determine the seroprevalence of T. gondii infection in dogs in Tehran, capital of Iran and to investigate possible associated risk factors.

Methods

Three hundreds ninety six serum samples were collected during 2007–8 from the dogs. Collected samples were tested using an indirect fluorescent antibody test (IFAT) in dilutions of 1:16 and more. All procedures were carried out in Shahrekord University, Iran. All the data were analyzed using SPSS software, qui square test with confidence interval of 0.95.

Results

From evaluated samples, 89 (22.47%) were positive in titers of at least 1:16. further evaluations in other dilutions showed positive results in dilutions of maximum 1:16, 1:32, 1:64, 1:128 and 1:256 in 38, 29, 15, 2 and 5 dogs respectively. Investigation of the role of risk factors showed no sex predisposition while infection rate was significantly higher in dogs older than one year old. Living places were of significant importance; infection rate was significantly higher in stray or guard dogs in compare with household dogs (P<0.05).

Conclusion

Relatively high seroprevalence of T. gondii infection in dogs in Tehran shows high environmental contamination. It is recommended that the dogs with suspected clinical signs be tested for T. gondii infection.     

Larva Migration and Eosinophilia in Mice Experimentally Infected With Gnathostoma spinigerum

Background

Gnathostoma spinigerum causes larva migran in human which is endemic in Southeast Asia. Information regarding larva migration is limited. In this study, we investigated the parasite migration by recovery of worms from the whole body of mouse after oral infection with advanced third stage larvae (AL3). The percentage of blood eosinophils was examined in parallel.

Methods

Mice were orally infected with AL3 and histological study of organs was investigated in order to study the migration of AL3, along with blood eosinophilia.

Results

At 1 hr post infection (PI), the larvae remained in the stomach, thereafter at 3, 5, 7, 10 and 24 hr PI; they were recovered from various organs including liver, mesentery, esophagus, diaphragm, lung, heart and dorsal fat. At day 15 PI, they were mostly found in muscles (76.47%). The average worm recovery (5 months) was 78.03%. The worms were found in the liver at every time point. Larva encystment was detected. There was a significant difference in blood eosinophils between the 8 larvae- (average 9.33% + 6.25%) and the 15 larvae-infected groups (average 22.66% + 11.03%). Surprisingly, the blood eosinophils (average 19.00% + 2.92%) were not higher in the higher infective dose- group (25 larvae).

Conclusion

Liver was involved by G. spinigerum throughout the study. We detected larva encystment which had never been reported in human gnathostomiasis. The highest percentage of eosinophil occurred during the invasive stage.     

Molecular and Parasitological Study of Cryptosporidium Isolates From Cattle in Ilam,West of Iran

Background

Cryptosporidiosis is one of the most important parasitic infections in human and animals. This study was designed for survey on the prevalence of Cryptosporidium infection in farms of Ilam, west of Iran, using parasitology method and genotyping by Nested PCR-RFLP.

Methods

Fecal samples of 217 cattle were collected fresh and directly from the rectum of cattle. All of the samples were examined by microscopic observation after staining with modified Ziehl-Neelsen (MZN). Genomic DNA extracted by using EURx DNA kit. A Nested PCR-RFLP protocol amplifying 825 bp fragment of 18s rRNA gene conducted to differentiate species and genotyping of the isolates using SspI and VspI as restriction enzymes.

Results

The prevalence of Cryptosporidium infection in cattle using both methods is 3.68%. Most of the positive cattle were calves under six months. Species diagnosis carried out by digesting the secondary PCR product with SspI that C. parvum generated 3 visible bands of 448, 247 and 106 bp and digested by VspI restriction enzyme generated 2 visible bands of 628 and 104bp. In this investigation all of the positive samples were Cryptosporidium parvum.

Conclusion

C. parvum (bovine genotype) detected in all positive cattle samples in Ilam, west of Iran. The results of the present study can help for public health care systems to prevention and management of cryptosporidiosis in cattle and the assessment of cattle cryptosporidiosis as a reservoir for the human infection.     

Helminth Infections in Rattus ratus and Rattus norvigicus in Tehran,Iran

Background

The aim of this study was to determine intestinal and liver helminth infections in Rattus rodents in Tehran Iran.

Methods

Overall, 306 traps were put in 39 different regions in Tehran from 2009 to 2010. Rodents, including R. rattus and R. norvegicus were caught by live-traps. They become unconscious and the spinal cords were cut, afterwards the body was dissected and the stomach, small intestine, large intestine, liver, and cecum were studied separately. The dominant type and the prevalence rate of parasites in the rodents were determined based on the infected parts of their body.

Results

After recognition of the helminthes’ types, among the 120 total number of rodents, 39 belonged to males, while among the infected rodents, 57(47.5%) were female and 18(15%) were male. The prevalence of infection in Tehran was 62.5%. Seventy cases (58.33%) of helminth infections were observed in R. rattus and 5 cases (4.16%) were observed in R. norvegicus. The maximum prevalence (15.5%) was seen in the center and east part of Tehran, while the minimum (9.16%) was in the north part of the city. The helminthes types and the corresponding percentages were Hymenolepis nana fraterna (35.8%), Heterakis spumosa (17.5%), Hymenolepis diminuta (7.5%) and Capillaria annulosa (1.6%). The dominant rodent was Rattus rattus and among the identified helminthes, Hymenolepis diminuta and Hymenolepis nana fraterna are zoonotic ones.

Conclusion

The information presented here improves our understanding of the major parasitic infections that rodents harbor and can transmit to human and animal populations in Iran. To prevent infectivity of human, the hazard of the identified zoonotic species needs to be contemplated.     

Epidemiological Aspects of Visceral Leishmaniasis in Baft District,Kerman Province,Southeast of Iran

Background

Visceral leishmaniasis (kala-azar) is an endemic disease in some areas of Iran. A cross- sectional study was conducted for sero-epidemiological survey of visceral leishmaniasis (VL) in Baft district from Kerman Province, southeast of Iran.

Methods

Blood samples were collected from children up to 12 years old and 10% of adult population from Baft villages with a multi-stage randomized cluster sampling. In addition, blood samples were collected from 30 domestic dogs from the same areas. All the collected blood samples were tested by direct agglutination test (DAT) for the detection of anti-Leishmania antibodies in both human and dog using the cut-off value of ≥1:3200 and ≥1:320, respectively. Parasitological, molecular, and pathological were performed on infected dogs. Chi-square and Fisher exact tests were used to compare sero-prevalence values.

Results

From 1476 collected human serum samples, 23 (1.55%) showed anti-Leishmania antibodies at titers of 1:800 and 1:1600 whereas 14 (0.95%) showed anti-Leishmania infantum antibodies at titers of ≥1:3200. No statistically significant difference was found between male (1.18%) and female (0.69%) sero-prevalence (P=0.330). Children of 5-8 years showed the highest sero-prevalence rate (3.22%). Seven out of 30 domestic dogs (23%) showed anti-Leishmania antibodies at titers ≥1:320. Leishmania infantum was identified in five infected dogs by nested – PCR assay.

Conclusion

It seems that visceral leishmaniasis is being endemic in southern villages of Baft district, southeast of Iran.     

Clinical Features and Laboratory Findings of Visceral Leishmaniasis in Children Referred To Children Medical Center Hospital,Tehran, Iran during 2004-2011

Background

Visceral leishmaniasis (VL) is one of the most important parasitic diseases endemic in northwestern and southern areas of Iran. The aim of the present study was to review the records of children hospitalized with VL in order to characterize the clinical features of children as well as laboratory finding in Children Medical Center Hospital, Tehran, Iran.

Methods

The medical records of all children with a final diagnosis of VL were reviewed from 2004 to 2011. Demographic, clinical information, laboratory finding and treatment were considered.

Results

A total number of 34 children with confirmed VL through 2004-2011 were included in the study. The most prevalent sign and symptoms were fever (97.1%), pallor and weakness (97.1%), appetite loss (61.8%), splenomegaly (97.1%) and hepatomegaly (88.2%). The most frequent laboratory abnormalities were hematological including anemia (97.1%), thrombocytopenia (91.2%) and leukopenia (67.6%). Direct agglutination test (DAT) was performed in 23 cases and all of them showed anti-Leishmania antibodies with titers of ≥ 1: 3200. In addition, 90% of patients had positive rK39 results. Identification of Leishmania in the aspirates of the bone marrow was found in 83.3% of patients.

Conclusion

Regional surveillance system in order to monitoring of leishmaniasis trends as well as detection of new emerging foci is recommended.     

Human Visceral Leishmaniasis in Kermanshah Province,Western Iran,During 2011-2012

Background

Visceral leishmaniasis (VL) or kala-azar is a parasitic disease caused by the species of Leishmania donovani complex. It is endemic in some parts of provinces of Iran. According to the reported cases of VL in Kermanshah Province in recent years, this study was conducted to determine the seroprevalence of VL in high risk villages of the province.

Methods

Totally, 1622 serum samples obtained from children under 15 years old and 178 from adults in 22 villages of studied areas. Serum samples were examined by direct agglutination test (DAT) for the detection of anti-Leishmania antibodies. Data were analyzed using SPSS software ver.11.5.

Results

Only 6 serum samples (0.33%) showed anti-Leishmania antibodies against L.infantum at titers ≥ 1/3200. Four of the seropositive cases had a history of kala-azar and Leishman bodies were seen in their bone marrows. The highest (0.5%) and lowest (0.29%) seroprevalence was seen in the age groups of 5-9 and 10-14 years old, respectively. None of the adults were seropositive. There were not any significant differences between the rate of seropositivity in males (0.36%) and females (0.31%). 66.7% of seropositive individuals showed clinical manifestations. The most important symptoms in Kala-azar patients were fever, hepato-spleenomegally and anemia.

Conclusion

Kala-azar is occurred sporadically in Kermanshah Province. But presence of significant number of positive sera confirms the necessity for attention of people and clinicians to kala-azar.     

Prevalence of Giardia duodenalis Infection in Household Cats of Ahvaz District,South-West of Iran

Background

The occurrence of Giardia duodenalis in cats is of potential significance from both clinical and public health perspectives. The object of this study was antigenic detection of G. duodenalis in household cats of Ahvaz district, South-West of Iran.

Methods

The prevalence of G. duodenalis was determined in fecal samples by two techniques: centrifugation-flotation and a commercial Giardia Antigen Test Kit (immunochromatography assay) in 150 household cats of different ages among referred cases to Veterinary Hospital of Ahvaz University from January 2008 to February 2010.

Results

Five out of 150 fecal samples (3.33%) were positive for antigen of G. duodenalis by immunochromatography assay. The prevalence was significantly higher in young cats less than 6 months (15.79%) compared with adult cats 6 months – 3 years (1.37%) (P=0.027) and above 3 years (1.72%) (P=0.044). The infection had more prevalence in diarrheic cats (17.39%) compared with non-diarrheic cats (0.79%) and the difference was significant (P=0.02) as well. The prevalence was higher in male cats (3.41%) than females (3.23%) and in the season of autumn (6.06%), but the difference was not significant between the prevalence of infection relative to host gender and season (P>0.05). Microscopy examination on fecal samples showed that 2% of the studied cats were positive.

Conclusion

The parasite antigen was present as a zoonotic infection in Ahvaz district, South-west of Iran. More sensitive techniques, such as immunochromatography assay, might yield more reliable results, in the detection of low levels of Giardia in fecal samples of cats.     

Epidemiological Aspects of Canine Dirofilariasis in the North of Iran

Background

Dirofilaria immitis is an important parasite in dog and other carnivores. Our objective was study on incidence and periodicity of heartworm in north of Iran and using other methods for its diagnosis in addition to Parasitology exam.

Methods

This survey spanned two years, between 2006 and 2008. Blood samples were collected from 431 stray dogs distributed along north of Iran, the coastal areas of the Caspian Sea. The Knott''s modified test was used for diagnosis of D. immitis and other filariae. Meanwhile, the periodicity of microfilaria in peripheral blood circulation was calculated and the imaging diagnosis techniques of four dogs that had positive results were done.

Results

Diagnostic parasitology results indicated that 16.01% of stray dogs were microfilaremic. Two different microfilariae were diagnosed: D. immitis in 13.69%, Dipetalonema reconditum in 1.86% and in 0.46% both of them. There was no statistically significant between infection to fiariae with sex and age of dogs. Also study on the periodicity of the presence of microfilaria in peripheral circulation showed that the highest rate of those was at 1 am and the lowest rate at 12 pm. Radiographic study showed distinctive signs with varied degrees of severity included: Tortuous and enlargement of main and lobar pulmonary artery, pulmonary parenchymal lesions and Right side heart enlargement that confirmed in electrocardiography. Also in echocardiographic images observed short parallel-sided images with the appearance of equal signs that indicated the presence of the heartworm.

Conclusion

These results showed that to obtain a reliable diagnosis of heartworm infection, imaging tests could support parasitological exams.     

Seroprevalence of Acanthamoeba Antibodies in Rheumatoid Arthritis Patients by IFAT,Tehran, Iran 2007

Background

This preliminary study was conducted to discriminate the prevalence of Acanthamoeba antibodies in rheumatoid arthritis (RA) patients and healthy controls to analyze the correlation between these two groups.

Methods

From October 2006 to August 2007 a total of 121 serum samples from RA patients attending the Rheumatolgy Department at Shariati Hospital in Tehran were obtained and stored at -20°C until using by indirect fluorescent-antibody test (IFAT). RA was diagnosed according to the American Collage of Rheumatology classification criteria. The organism used in this study was isolated from various water resources in Tehran, Iran cultured axenically and then went on a PCR assay based on 18S rRNA to identify the genus Acanthomoeba. Indirect immunofluorescence antibody (IFA) staining of serum samples was carried out to detect anti Acanthomoeba antibodies.

Results

In culture, out of 22 samples, 13(59%) were grown in xenic but only two in axenic medium. PCR amplified a 904bp fragment, specific for Acanthamoeba. Of examined serum samples, Acanthamoeba antibodies were present in 70 (57.8%) and 52 (41.2%), respectively. The highest titer of antibodies (1:320) was detected in one patient with RA.

Conclusion

Our study supports the hypothesis that some parasitic microorganisms can involve and contribute toward the development of rheumatoid syndromes.     

First Molecular Identification of Sarcocystis ovicanis (Protozoa,Apicomplexa) in the Brain of Sheep in Iran

Background

The objective of the present study was to survey the presence of Sarcocystis in sheep’s brain in North Khorasan Province.

Methods

In general, 80 samples of sheep’s brain were collected from slaughtered sheep in slaughterhouses of North Khorasan Province. Tissue digestion method was used for observing bradyzoites in tissues. Histopathological processing tracing Sarcocystis and ensuing structural change in the brain tissue were conducted. PCR analysis was conducted on all the brain samples. Sequencing was done for one PCR product. Genotype was identified by Blast search and homology analysis.

Result

Sarcocystis spp. was found in one of the brain samples (1.25%) using tissue digestion method. The presence of bradyzoite was also confirmed in the prepared histopathological sections. PCR analysis was positive in one of samples. Genotyping of one sample proved that Sarcocystis species was Sarcocystis ovicanis and the nucleotide sequence of this parasite was deposited in the GenBank database under accession number No.{"type":"entrez-nucleotide","attrs":{"text":"KF489431","term_id":"540198437","term_text":"KF489431"}}KF489431.

Conclusion

Sarcocystis ovicanis can involve brain tissue of sheep and consequently causes clinical symptoms.