High Seroprevalence of Toxoplasma gondii Antibody in HIV/AIDS Individuals from North of Iran

Background:

Toxoplasmosis in immunocompetent people is generally asymptomatic but in immunocompromised patients including HIV/AIDS, cancer patients, and organ transplant recipients, etc. it can lead to serious pathological problems. The objective of current study was to determine the seroprevalence of T. gondii IgG and IgM antibodies in HIV/AIDS patients using ELISA technique in Mazandaran Province, northern Iran.

Methods:

Overall, 82 serum samples (61 males and 21 females) were collected from HIV/AIDS patients in Mazandaran Provinces, in 2013. Sera were surveyed employing ELISA assay. Data were analyzed using Chi-Square or Fisher exact test. In addition, before sampling a questionnaire was filled out for each subject.

Results:

Overall seroprevalence of examined sera was 96.3% for IgG antibody but none of the sera shown IgM antibody against T. gondii. The seroprevalence of toxoplasmosis in males and females was 96.7% and 95.2%, respectively. An antibody titer of >1 IU/ml was considered as positive. Furthermore, none of the included variables statistically was significant.

Conclusions:

Seroprevalence of chronic (latent) toxoplasmosis in HIV/AIDS patients in Mazandaran Province is high compared to toxoplasmosis in general population. Consequently, the risk of acquiring Toxoplasma encephalitis in examined seropositive HIV/AIDS patients of Toxoplasma is high.     

Morphological and Molecular Survey of Naegleria spp. in Water Bodies Used for Recreational Purposes in Rasht city,Northern Iran

Background:

Naegleria spp. is a free-living amoeba of which some species including N. fowleri and N. australeinsis are highly pathogenic in human and animals. These widespread amoebae could be found in different environmental sources particularly in aquatic resources of tropical and subtropical regions. The most important source of infection is via recreational water contact. Due to the lack of thorough research regarding species of Naegleria spp. in aquatic sources, the present study was conducted.

Methods:

In the present study, 60 samples were collected from recreational water resources of Rasht city, Guilan province, north of Iran. After filtering and culturing the samples, plates were examined by microscopic method and according to the page criteria. DNA of vahlkampfiid-positive samples were then extracted using phenol-chlorophorm method. Amoebae genus was identified by targeting the ITS-region and sequencing based-approaches.

Results:

Nine (15%) samples out of a 60 total samples were positive for Naegleria spp. of which seven belonged to potentially pathogenic N. australiensis. Two other strains were belonged to non-pathogenic N. pagei.

Conclusion:

The present research was the first report of occurrence of N. australiensis and N. pagei in Rasht city, north Iran. This study reflects the occurrence of Naegleria spp. in water sources of Guilan Province, Iran.     

Identification of Leishmania Species Using PCR Assay on Giemsa-Stained Slides Prepared From Cutaneous Leishmaniasis Patients

Background

Leishmaniasis is a group of diseases that are created by intracellular parasites of Leishmania. Cutaneous leishmaniasis is considered as one of the health problems in some provinces of Iran.

Methods

In this study, a total of 178 Giemsa-stained slides from confirmed cases of cutaneous leishmaniasis were examined. The slides were prepared from the patients with cutaneous leishmaniasis that referred to health centers and infected during the epidemic of cutaneous leishmaniasis in Poldokhtar city, Lorestan Province, Iran in 2006.Genomic DNA from each slide was extracted. After DNA extraction, ITS-PCR was used.

Results

Out of 178 slides, 129 (72.47%) samples had a band in the range of 485 bp and 49 (27.53%) samples 626 bp that matched L. tropica and L. major standard samples, respectively.

Conclusion

This study showed that Leishmania DNA could be efficiently extracted and amplified even from old Giemsa-stained microscopic slides that were stored more than 6 yr. In this study was shown that both L. tropica and L. major species exist in Lorestan Province.     

Molecular Identification of Leishmania Species Using Samples Obtained from Negative Stained Smears

Background

Cutaneous Leishmaniasis (CL) is a parasitic skin disease. Diagnosis primarily is based on clinical signs and microscopic observation of parasite on direct stained smears or tissue sections. Sensitivity of direct smear is not as high as molecular methods. The aim of this study was to identify and characterize Leishmania species among the negative direct smears obtained from skin ulcers suspected to CL by PCR method.

Methods

Among 81 patients with suspicious skin lesions to CL referred to the Parasitology lab, negative Giemsa stained smears were collected. DNA extraction performed by scraping stained smears, then PCR was performed.

Results

Among the DNA extracted from smears, L. tropica was isolated from 9 (11.1%) of the smears and L.major was not isolated from any samples.

Conclusion

Direct microscopy on stained smears for diagnosis of leishmaniasis is not enough accurate. PCR is recommended for clinically suspected lesions with negative result of direct smear.     

Human Hydatidosis/Echinococosis in North Eastern Iran from 2003–2012

Background:

Human cystic echinococcosis (hydatidosis) continues to be an essential cause of morbidity and mortality in many parts of the world.

Methods:

We studied hydatid cyst pattern in hospitalized adult patients from 2003 to 2012 in Mashhad and Neyshabour, northeast of Iran.

Results:

Overall, 1342 patients, 711 females (53%) and 631 males (47%) diagnosed as infected with hydatid cyst were evaluated. Their age was between 1 and 91 yr (mean age 37.75). The most affected age group was 20–30 yr old. Totally, 953 cases (71%) were urban and 375 cases (27.8%) were rural residents. The most common localization of cysts was the liver and lung. The housewives were the most frequently infected occupations.

Conclusion:

The rate of infection with hydatid cyst is high in Mashhad, northeast of Iran, and the incidence of human hydatidosis tends to increase in recent years so control and prevention programs are recommended.     

Pathogenic Free-Living Amoebae Isolated From Contact Lenses of Keratitis Patients

Background:

Free-living amoeba (FLA)-related keratitis is a progressive infection of the cornea with poor prognosis. The present study aimed to investigates the contact lenses of patients with keratitis for pathogenic free-living amoebae.

Methods:

Overall, 62 contact lenses and their paraphernalia of patients with keratitis cultured and tested for the presence of free-living amoebae using morphological criteria. Unusual plates including plates containing mix amoebae and Vermamoeba were submitted to molecular analysis.

Results:

Out of 62 plates, 11 revealed the outgrowth of free living amoeba of which 9 were Acanthamoeba, one plates contained mix amoebae including Acanthamoeba and Vermamoeba and one showed the presence of Vermamoeba. These two latter plates belonged to patients suffered from unilateral keratitis due to the misused of soft contact lenses. One of the patients had mix infection of Acanthamoeba (T4) and V. vermiformis meanwhile the other patient was infected with the V. vermiformis.

Conclusion:

Amoebic keratitis continues to rise in Iran and worldwide. To date, various genera of free-living amoebae such as Vermamoeba could be the causative agent of keratitis. Soft contact lens wearers are the most affected patients in the country, thus awareness of high-risk people for preventing free-living amoebae related keratitis is of utmost importance.     

Mini-Exon Genotyping of Leishmania Species in Khuzestan Province,Southwest Iran

Background

Leishmaniasis is a protozoan disease cause by Leishmania genus. Anthroponotic and zoonotic cutaneous leishmaniasis are endemic in Iran. The aim of this study was to identify the causative agent of cutaneous leishmaniasis by mini-exon gene in five regions of Khuzestan Province, southwest of Iran.

Methods

From 2007 to 2008 in this cross-sectional study, cutaneous samples were collected from patients referred to Health Centers and Hospitals of the Khuzestan Province for cutaneous leishmaniasis diagnosis and cultured in Novy-MacNeal-Nicolle (NNN) and RPMI 1640. The propagated promastigotes were harvested and Leishmania species of cutaneous leishmaniasis were identified by RFLP and DNA sequencing of the PCR generated fragments.

Results

L. major and L. tropica were the causative agents of cutaneous leishmaniasis by predominantly of L. major species. The alignment of the mini-exon sequencing isolates with reported sequencing of L. major and L. tropica revealed 92%-99% identity.

Conclusion

Our study showed that mini-exon PCR-RFLP was useful method to identify the causative species of cutaneous leishmaniasis.     

Seroprevalence and Risk Factors of Toxoplasma gondii Infection in Buffaloes,Sheep and Goats in Yunnan Province,Southwestern China

Background:

The seroprevalence of Toxoplasma gondii infection in buffaloes, sheep and goats in Yunnan Province, southwestern China was conducted between May 2012 and December 2013.

Methods:

A total of 973 (427 buffaloes, 154 sheep and 392 goats) serum samples were collected from seven administrative regions of Yunnan Province, and examined for T. gondii antibodies by indirect hemagglutination (IHA) test. Some risk factors related to species, age, gender and geographical origin were determined using a multinomial logistic regression.

Results:

The overall seroprevalence of T. gondii in ruminant species was estimated at 11.9%. The final logistic regression model demonstrated that host species and geographical origin were the main risk factors associated with T. gondii infection (P<0.05).

Conclusion:

Taken together, the results of the present study revealed a high exposure to T. gondii in ruminant species in Yunnan Province, which has an important implication for public health.     

Immunoproteomic Analysis of the Excretory-Secretory Proteins from Spirometra mansoni Sparganum

Background

Sparganosis is caused by the invasion of Spirometra sparganum into various tissues/organs. Subcutaneous sparganosis can be diagnosed by biopsy, while visceral/cerebral sparganosis is not easy to be diagnosed. The diagnosis depends largely on the detection of specific anti-sparganum antibodies. The specificity of the ELISA could be increased by using S. mansoni sparganum excretory–secretory (ES) antigens, but it also had the cross-reactions with sera of patients with cysticercosis or paragonimiasis. The aim of this study was to identify early specific diagnostic antigens in S. mansoni sparganum ES proteins.

Methods

The sparganum ES proteins were analyzed by two-dimensional electrophoresis (2-DE) and Western blot probed with early sera from infected mice at 14 days post-infection. The immunoreactive protein spots were characterized by MALDI-TOF/ TOF-MS.

Results

A total of approximately 149 proteins spots were detected with isoelectric point (pI) varying from 3 to 7.5 and molecular weight from 20 to 115 kDa and seven protein spots with molecular weight of 23-31 kDa were recognized by the infection sera. Three of seven spots were successfully identified and characterized as the same S. mansoni protein (cysteine protease), and the proteins of other 4 spots were not included in the databases.

Conclusion

The cysteine protease from S. mansoni ES proteins recognized by early infection sera might be the early diagnostic antigens for sparganosis.     

Canine Visceral Leishmaniasis in Boyer Ahmad District,Kohgiluyeh & Boyer Ahmad Province,Southwest of Iran

Background

Mediterranean type of visceral leishmaniasis (VL) is present in different parts of Iran. Several studies have identified dogs as the main reservoirs of the VL caused by Leishmania infantum in Iran and other Mediterranean regions. This study aimed to determine the seroprevalence of canine visceral leishmaniasis as animal reservoir host for human visceral leishmaniasis in Boyer Ahmad district in southwest of Iran.

Methods

A seroepidemiological study was carried out to determine the seroprevalence of canine visceral leishmaniasis (CVL) among ownership dogs by using direct agglutination test (DAT) in 23 of 182 villages of Boyer Ahmad district, during August 2009 to August 2010. One hundred and seventy serum samples from ownership dogs were selected by multi-stage cluster sampling in villages of Boyer Ahmad district. All samples were tested by DAT and anti-Leishmania antibodies titers at ≥ 1:320 was considered as positive.

Results

Of the 170 serum samples, 10% were positive by DAT at titers of 1:320 and higher. No statistical significant difference was found between male (10.7%) and female (8.3%) seroprevalence. The highest seroprevalence rate (15.1%) was observed among the ownership dogs of four to seven years age. Altogether, seventeen (25.4%) of the seropositive dogs had clinical signs and symptoms.

Conclusion

It seems that Boyer Ahmad district is an endemic area for canine visceral leishmaniasis in Iran.     

DNA Sequence Polymorphism of the Lactate Dehydrogenase Genefrom Iranian Plasmodium vivax and Plasmodium falciparum Isolates

Background:

Parasite lactate dehydrogenase (pLDH) is extensively employed as malaria rapid diagnostic tests (RDTs). Moreover, it is a well-known drug target candidate. However, the genetic diversity of this gene might influence performance of RDT kits and its drug target candidacy. This study aimed to determine polymorphism of pLDH gene from Iranian isolates of P. vivax and P. falciparum.

Methods:

Genomic DNA was extracted from whole blood of microscopically confirmed P. vivax and P. falciparum infected patients. pLDH gene of P. falciparum and P. vivax was amplified using conventional PCR from 43 symptomatic malaria patients from Sistan and Baluchistan Province, Southeast Iran from 2012 to 2013.

Results:

Sequence analysis of 15 P. vivax LDH showed fourteen had 100% identity with P. vivax Sal-1 and Belem strains. Two nucleotide substitutions were detected with only one resulted in amino acid change. Analysis of P. falciparum LDH sequences showed six of the seven sequences had 100% homology with P. falciparum 3D7 and Mzr-1. Moreover, PfLDH displayed three nucleotide changes that resulted in changing only one amino acid. PvLDH and PfLDH showed 75%–76% nucleotide and 90.4%–90.76% amino acid homology.

Conclusion:

pLDH gene from Iranian P. falciparum and P. vivax isolates displayed 98.8–100% homology with 1–3 nucleotide substitutions. This indicated this gene was relatively conserved. Additional studies can be done weather this genetic variation can influence the performance of pLDH based RDTs or not.     

Epidemiology,Pathology and Treatment of Cutaneous Leishmaniasis in Taif Region of Saudi Arabia

Background

Cutaneous leishmaniasis is an annoying and disfiguring disease affecting around 1,500,000 individuals globally. There are endemic pockets of this disease in Taif region. In some patients, lesion often weeps and leads to scar formation. The study was conducted to see the efficacy of fluconazole and itraconazole in the patients of cutaneous leishmaniasis and the effect of these drugs on liver enzymes and kidney markers.

Methods

Positivity of Leishmania was recorded by microscopic examinations of smears. Specific diagnosis for Leishmania major and L. tropica was made with the help of nested polymerase chain reaction. Fluconazole was given at the rate of 200mg/day while itraconazole was given at 150mg/day for six weeks. AST, ALT, creatinine and urea were estimated during medication.

Results

Leishmania major and L. tropica were the species responsible for cutaneous leishmaniasis in Taif region. 81% patients had single lesions, mostly on face followed by hands and legs. 15% of the lesions had bacterial contamination. In terms of efficacy, fluconazole gave slightly better results compared to itraconazole. After 6 weeks of medications, slightly elevated values were recorded for liver enzymes and creatinine.

Conclusion

Transmission of leishmaniasis in Taif region is probably because of poor coverage of residual insecticides spraying at hiding places in pile-ups of rocks and abandoned houses from where sand flies visit nearby houses and cattle sheds during night. Fluconazole and itraconazole may be used for the treatment of cutaneous leishmaniasis with good recovery rate and fewer side effects.     

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.     

In vivo Susceptibility of Plasmodium vivax to Chloroquine in Southeastern Iran

Background

Plasmodium vivax is the predominant species causes of malaria with about 90% total annual reported malaria in Iran. This study conducted to determine the susceptibility of Plasmodium vivax isolates to chloroquine in Sistan and Balochistan Province, southeastern Iran.

Methods

A total 270 subjects with symptomatic malaria and confirmed P. vivax infection completed the designed 28-day in vivo study. The thick and thin film blood smears were screened for malaria parasites by microscopy. The nested PCR was applied using the Plasmodium 18 subunit ribosomal ribonucleic (Ssr RNA) genes for detecting mixed infections and diagnosis of parasites in the samples with low parasite on days 0, 5, 6, 7, and 28.

Results

P. vivax was cleared in 15%, 50%, 95%, and 100% of patients on days 1, 2, 3, 4 respectively by microscopy assessment. Six patients were exhibited specific P. vivax band in nested PCR on day 5. No recurrence was observed on days 7, 14 and 28. Mean (±standard deviation) parasite clearance time was 2.41 (±0.8) days.

Conclusion

P. vivax is still susceptible to chloroquine in Southeatern Iran. This finding is compatible with results of neighboring countries Pakistan and Afghanistan.     

Leishmaniasis in Turkey: Determination of Leishmania Species by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS)

Background

Cutaneous leishmaniasis (CL) is endemic in Southeastern Anatolia, mainly in Sanliurfa and Hatay provinces, and the causative agents are mostly Leishmania tropica and less frequently L. infantum. Here, we report the first MALDI-TOF analyses of Leishmania promastigotes obtained from the cultures of two CL cases from Osmaniye and Hatay provinces who were initially diagnosed by microscopy, culture and identified as L. infantum with Real-Time PCR (RT-PCR).

Methods

Samples obtained from the skin lesions of patients were initially stained with Giemsa and cultivated in NNN medium. Examination of the smears and cultures revealed Leishmania amastigotes and promastigotes, respectively. The promastigotes (MHOM/TR/2012/CBU15 and MHOM/TR/2012/MK05) obtained from the cultures of both patients were used for RT-PCR targeting the ITS-1 region in the SSU of rRNA. The reference strains of four Leishmania species (L. infantum, L. donovani, L. tropica and L. major) were initially assessed with MALDI-TOF and their data were added to MALDI-TOF Biotyper Library.

Results

Both RT-PCR and MALDI-TOF analyses indicated that the causative agent in both patient samples was L. infantum.

Conclusion

Despite disadvantages such as requirement of culture fluid with nothing but promastigotes and high cost, MALDI-TOF analysis may be a fast, sensitive and specific diagnostic tool in especially large-scale research studies, where the cost declines, relatively.     

Evaluation of Recombinant SAG1 Protein for Detection of Toxoplasma gondii Specific Immunoglobulin M by ELISA Test

Background

Toxoplasmosis is a serious disease in immunocompromised patients and pregnant women. Differentiation of acute and chronic infection is a major challenge in serodiagnosis of the disease. Since the aim of this study was to assess the diagnostic utility of recombinant SAG1 (rec-SAG1) for the detection of Toxoplasma-specific IgM antibodies in human sera, by an enzyme-linked immunosorbent assay (ELISA).

Methods

The purified recombinant protein SAG1 was applied in house ELISA test and the ability of it in binding to specific immunoglobulin M in 30 serum samples of acute infected patients was evaluated. The results obtained by assays with the recombinant SAG1 and standard commercial assays were compared.

Results

The sensitivity and specificity of in house ELISA compared to a standard commercial ELISA (com-ELISA) were 80% and 90%, respectively.

Conclusion

It was concluded that the rec-SAG1 could be an alternative marker for detection of anti Toxoplasma-specific IgM and diagnosis of acute infection.     

Seroprevalence of Toxoplasma gondii Infections in Pregnant Women in Gorgan City,Golestan Province,Northern Iran-2012

Background

Toxoplasma gondii is one of the most prevalent parasites of human and warm- blooded animals. Toxoplasmosis is important especially in two groups: pregnant women and immunocompromised patients. If women acquire the primary infection during the pregnancy, it would be life threatening or remains severe disorders for the fetus. This study was performed to evaluate the seroprevalence of T. gondii infection in pregnant women referred to Health Center in Gorgan City, Golestan Province, northern Iran.

Methods

Serum samples were collected from pregnant women referred to Health Center in Gorgan City, south eastern Caspian Sea. Anti- Toxoplasma IgG and IgM antibodies were determined by commercially ELISA kits and the relation of infection with socio-demographic and risk factors such as age, education, occupation, cat ownership, soil contact and some other factors was studied.

Results

From 555 tested sera of pregnant women referred to Health Center in Gorgan, 39.8% had IgG antibodies against T. gondii and 3.4% were positive for IgM antibodies. A significant correlation was seen between T. gondii infection with age and soil contact.

Conclusion

About 60% of pregnant women in Gorgan City are seronegative against T. gondii, so they should considered as at risk persons.