Iranian Journal of Parasitology
Volume:19 Issue: 1, Jan-Mar 2024

Uncovering the roles and characteristics of pathogenesis-related molecules can help us develop novel management methods in parasitology. In this study, we studied the expression levels of Strongyloides stercoralis heat shock protein70 (HSP70) (Sst-hsp-70) and astacin (Sst-ast) as pathogenesis-related genes as well as the expression of S. ratti HSP70 and HSP17.1 (Sra-hsp-70, Sra-hsp-17.1) in the larvae and adult stages of S. stercoralis.

A hyperinfection isolate of S. stercoralis from Gilan Province, northern Iran was cultivated on nutrient agar. After a couple of days, parasites in different stages of life were collected, and total RNA was extracted. The expression levels of astacin and HSP genes were compared by real-time PCR.

Statistically higher expression levels of Sst-ast, Sst-hsp-70, and Sra-hsp-70 genes in L3 larvae than in adults were observed. However, the expression level of Sra-hsp-17.1 was non-significantly lower in the larval stage than in adult worms.

Higher expression levels of Sst-ast, Sst-hsp-70, and Sra-hsp-70 genes in the larval stages of S. stercoralis suggest the potential role of these enzymes in parasite cutaneous invasion and pathogenesis. However, higher expression of Sra-hsp-17.1 in adult forms is probably involved in resistance and survival mechanisms. The similarity in gene expression between S. stercoralis and S. ratti can provide helpful hints to better understand strongyloidiasis from various perspectives, including pathogenesis, proper diagnosis, and targeted treatment.

We aimed to determine the prevalence of Giardia intestinalis and Cryptosporidium spp. in patients who admitted hospital with diarrhea and to gain information about the transmission of these parasites in Ağrı, Türkiye.

This study included 184 patients who applied to Ağrı-Diyadin State Hospital, Türkiye in 2022. The immunochromatographic card test was used for detection of the G. intestinalis and Cryptosporidium spp. Nested PCR-RFLP analysis of the COWP gene and sequence analysis of the gp60 gene were used to genotype and subtype Cryptosporidium spp., whereas Nested PCR and sequence analyses of β-giardin gene were used genotype G. intestinalis.

Of the 184 stool specimens examined, 12 (14.29%) and 7 (3.80%) were positive for G. intestinalis and Cryptosporidium spp., respectively. The Cryptosporidium species were identified as C. parvum belonging to the IId subtype family. The G. intestinalis were identified assemblages A.

Assemblage A, which is associated with diarrhea, is responsible for giardiasis and C. parvum IId subtype, often found in sheep, goats and cattle, is responsible for cryptosporidiosis in Ağrı, Türkiye.

Trichomoniasis is one of the most common sexually transmitted infections worldwide. The growing concern of drug resistance of this infection has cautioned the need for new drug development. We evaluated the potential antiproliferative and apoptotic effect of α-pinene and tannic acid (TA) on Trichomonas vaginalis cells. In addition, the cytotoxicity of agents on Vero cells was investigated.

  Trichomonas cells were axenically cultured in TYI-S-33 medium. In vitro antiproliferative activity of α-pinene, TA, and metronidazole was investigated against Trichomonas cells. The assays were carried out in triplicate using microtiter plate and trypan blue staining method. Annexin V/PI staining with flow cytometry was used to evaluate apoptosis induction. In addition, the cytotoxic effect was measured by MTT assay.

α-Pinene and TA exhibited significant inhibition of the Trichomonas cells and the lowest IC50 values were 22.9 µg/ml and 140 µg/ml at 48 hours’ incubation, respectively. The CC50 was found at 116 μg/ml for α-pinene and 473 μg/ml for TA, after 48 hours of treatment. The flow cytometry study demonstrated that the natural compounds induced apoptosis in Trichomonas cells. After 24 hours of treatment, the induction of apoptosis was 5.2% - 36.6% at concentrations of 3.9 - 62.5 μg/ml for α-pinene and TA induced-apoptosis was 6.1% - 53.8% at concentrations of 125-2000 μg/ml.

Although the results show the antiproliferative and apoptotic effect of α-pinene and TA on Trichomonas cells, in vivo studies are needed to further clarify the effects of these compounds.

Black disease, also known as visceral leishmaniasis (VL), is a parasitic illness caused by various Leishmania species. The risk of morbidity and mortality increases with delayed diagnosis and treatment. Early VL diagnosis and fast appropriate treatment are critical issues in endemic areas.  

This study was a retrospective cross-sectional study to investigate the diagnostic and therapeutic course of patients admitted with the diagnosis of VL in the Children's Medical Center (CMC) Hospital, Tehran, Iran. All cases of VL in patients under the age of 18 hospitalized between the years 2012 to 2022 were enrolled.

Twenty-seven patients were enrolled with an average age of 28.13 months with the majority of females (51.8%). Common clinical signs were fever (96.2%) and splenomegaly (92.59%). However, lymphadenopathy was rare. The largest number of patients was from Tehran Province, followed by Ardabil, Khuzestan, Gilan, and Alborz provinces. The most common hematological abnormalities were anemia (85.1%) and thrombocytopenia (44.4%). In accordance with the treatment strategy, liposomal amphotericin B and amphotericin B deoxycholate were given to 11 and 5 patients, respectively. Eleven of them received glucantime. The average length of hospitalization for liposomal amphotericin B was 15.36 ± 12.49 days. In comparison with glucantime (18.38 ±10.26 days) and amphotericin B deoxycholate (20.20± 6.18 days), liposomal amphotericin B group hospitalization was shorter than others were.

VL should be included in the differential diagnosis of any child who presents with fever, splenomegaly, and anemia. Concerning the treatment strategy in this study, liposomal amphotericin B had more efficiency and shorter hospitalization duration.

Toxocara canis is one of the most important causes of animal toxocariasis with global distribution. We aimed to find out the seroprevalence of toxocariasis in dogs in a rural area in Fars Province, south of Iran.

Dogs blood samples were collected from 60 dogs in three rural areas in the Sar Mashhad region, Fars Province. Dog sera were evaluated for anti-Toxocara antibodies by an indirect ELISA method. The association between the seropositivity and age, gender, and the sampling location were statistically evaluated.

Serological assay detected anti-Toxocara antibodies in sera of 32 out of 60 dogs, corresponding to a seroprevalence of 53.3%. The rate of seropositivity was higher in the male dogs.  The rate of seropositivity was higher in old dogs. This rate increased with increasing age, however, the association between age and Toxocara seropositivity was not statistically significant.

The high prevalence of Toxocara infection in dogs in the current study area confirms that infected dogs are an important source of Toxocara infection for their owners and people who are in close contact with these animals, especially children.

Hydatid cyst is a parasitic infection, often caused by Echinococcus granulosus. Although it is classified as a benign disease, cyst ruptures in the abdomen can be fatal. Ruptures occur spontaneously or after trauma. We aimed to report data from patients who underwent emergency surgery due to spontaneous intra-abdominal hydatid cyst rupture.

Upon a retrospective review of the records at Department of General Surgery, Faculty of Medicine, Harran University, Şanlıurfa, Turkey, we found that 34 cases were operated on due to hydatid cyst rupture between January 2012 and October 2022. All patients were operated on in an emergency, and partial cystectomy, intra-abdominal irrigation, and drainage were performed using laparotomy. The patients were evaluated in terms of age, sex, symptoms, radiological findings, laboratory results, intraoperative findings, and postoperative follow-ups.

Twenty-two (64.7%) female and 12 (35.3%) male patients were enrolled. The mean age was 39.1 (±17.58) years. All patients experienced spontaneous rupture. The ruptured cyst was found in the liver in 32 patients (94%), the spleen in 1 patient (3%), and the pelvis in 1 patient (3%). The diagnosis was determined using ultrasonography in 12 (35.3%) patients, computed tomography in 21 (61.8%) patients, and magnetic resonance imaging in 1 (2.9%) patient. All patients exhibited acute abdomen and leukocytosis. The average length of hospital stay was 5.14 (±1.37) days.

Hydatid cyst rupture should be considered in cases of acute abdomen, particularly in regions where the disease is endemic, as in our region. The ruptured hydatid cyst was primarily observed in the liver (94.11% of cases).

We aimed to investigate the cytotoxic and apoptotic effects of miltefosine on Toxoplasma gondii RH strain by various techniques.

The study was conducted at the Department of Parasitology and Mycology, Urmia University of Medical Sciences, Iran in 2020. Four groups of five BALB/c mice were selected. The cytotoxicity test was conducted by adding miltefosine to T. gondii tachyzoites; control tachyzoites received PBS and MTT assay was done on each suspension. For evaluating the Th1-type immune responses, the serum levels of IFN-γ and nitric oxide (NO) were assessed in mice after injecting tachyzoites and miltefosine, respectively. The flow cytometry technique was performed on T. gondii tachyzoites challenged with IC50 and IC90 doses of miltefosine and unchallenged cells. DNA fragments in T. gondii tachyzoites were detected by Terminal dUTPnick-end labeling (TUNEL) method.

Overall, 256, 64, 32, and 16 µg concentrations of miltefosine, respectively could kill more than 50% of viable T. gondii tachyzoites. The infected mice group, treated with miltefosine, significantly produced more IFN-γ relative to other groups (P< 0.001). Moreover, a significant difference was found in inducible NO synthase between the experimental and control groups (P<0.05). The flow cytometry results demonstrated a concentration-dependent apoptosis rate in tachyzoites incubated with miltefosine, though the necrosis rate was non-significant. DNA fragmentation analysis indicated oligonucleotides (18-200 bp) in tachyzoites treated with 11µg of miltefosine for 24, 48 and 72 h. However, this pattern was not observed in untreated control microorganisms.

Miltefosine could be a favorable candidate for use as a new treatment for toxoplasmosis.

The current in silico study was done to determine the primary biochemical features and immunogenic epitopes of Echinococcus granulosus glutathione S-transferase protein as a potential vaccine candidate.

Several web tools were employed to predict physico-chemical properties, antigenicity, allergenicity, solubility, post-translational modification (PTM) sites, subcellular localization, signal peptide, transmembrane domain, secondary and tertiary structure followed by refinement and validations. In addition, B-cell epitopes were predicted and were screened using various web servers, while MHC-binding and CTL epitopes were predicted using IEDB and NetCTL servers, respectively.

The protein had 219 residues with a molecular weight of 25.55 kDa and alkaline isoelectric pH (7.5). This protein was stable, thermotolerant (aliphatic index: 78.04) and hydrophilic (GRAVY: -0.440). The predicted antigenicity scores were low and the protein was non-allergenic in nature. There were no transmembrane domain and signal peptide in the sequence. Moreover, several B-cell, MHC-binding and CTL epitopes were found in the EgGST protein, which could be further used in multi-epitope vaccines.

Further studies are needed on the development of vaccines in vivo using EgGST alone or in combination with other antigens in the future.

The production of bovine theileriosis vaccine involves in vitro cultivation of Theileria annulata schizont-infected cell lines. Fetal bovine serum (FBS) is commonly used in animal cell culture, including the Theileria cell line. However, we aimed to reduce the amount of serum needed for cell culture by modifying the Stoker culture medium with supplements such as excretion factor and serum substitutes.  

To evaluate the effectiveness of these modifications, techniques such as cell counting, cell viability assays, and genomic analysis were employed in the Parasitic Vaccines Production Department of Razi institute of Iran, from 2020 to 2022.  Statistical analysis was used to compare the results of different experimental conditions.  

The three experimental media were as effective as the commonly used 10% Stoker medium in supporting the growth and viability of cells.

The significant reduction in the required amount of serum and the remarkable cell growth achieved by using defined serum replacements for the production of cell culture media is a significant step towards the preparation of a proper cell culture medium for the production of bovine Theileriosis vaccine.

Bovine babesiosis, a global disease, has not been studied so far in Kashmir valley, which is having temperate type of climate as compared to rest of India having tropical to sub-tropical climate, so we felt the need to investigate it.

To diagnose the babesiosis in clinically suspected cattle (n=450), peripheral blood film examination and PCR tests using generic and species-specific primers targeting Babesia/Theleria genera and   B. bigemina, B. bovis as well as B. divergens, respectively were conducted. Four PCR products were sequenced and subjected to BLASTn analysis. Ticks were collected from the clinically suspected animals and identified as per the standard morphological keys.

The prevalence of babesiosis among suspected cattle in central Kashmir by peripheral blood film examination and PCR technique was 11.11% and 33.62%, respectively. The 18S rRNA gene of Isolate B1 of Babesia spp. showed 99.0 to 100% nucleotide sequence homology with 18S rRNA gene of different isolates of B. bigemina registered in the GenBank, while as 18S rRNA gene of Isolate Z showed 98.5 to 99.2% and 93.1 to 93.9% nucleotide sequence homology with 18S rRNA gene of different isolates of Babesia spp. and B. bigemina, respectively, registered in the GenBank. Rhipicephalus spp. and Haemaphysalis spp. were the two major tick genera identified in the present study.

Bovine Babesiosis in Kashmir is attributed to B. bovis,  B. bigemina and some other Babesia spp. or strains which needs further investigation. To our knowledge, this is the first report of Babesia bovis from northern India in cattle.

To investigate Molecular Genotyping of Giardia duodenalis in Humans in the Yazd County, Central of Iran.

Total of 35 fecal samples were collected from patients referred to Yazd central laboratory from February to July 2022. All the samples were included in this study after microscopic observation of G duodenalis. DNA samples were extracted using related kit and were analyzed by Nano Drop. The molecular assessment was carried out using semi-nested PCR using the target gene of gdh. All amplified samples were sequenced using Sanger method. The sequences were analyzed by BLAST for assemblage identification.    

Out of 35 samples, 24 (68.57%) and 11 (31.43%) were male and female, respectively. All included samples were amplified using the specific gdh primer pair. The molecular analysis showed 17 isolates (48.57%) as assemblage BIV, 8 isolates (22.86%) as assemblage BIII, 6 isolates (17.14%) as assemblage AII and 4 isolates (11.43%) as assemblage AIII (p<0.05).

The molecular analysis conducted in this study showed that assemblages A and B are the most prevalent in Central Iran. The molecular identification of G duodenalis isolates from animals and implementing control programs is strongly recommended as a preventive measure to limit the transmission of giardiasis to humans.

Intestinal parasitic infections are still a considerable global public health problem. We aimed to determine the frequency of intestinal parasitic infections among people referring to the central laboratory of Meshkin Shahr City, Ardabil Province, Iran.

In this cross-sectional survey, 460 fecal samples were collected randomly from persons referred to the central laboratory of Meshkin Shahr City, from January to June 2022. The samples were examined by direct wetmount, Trichrome and modified Ziehl-Neelsen staining, formalin ethyl acetate sedimentation, and agar plate culture.

The frequency of intestinal parasites was 15.7% (72 out of 460 cases), with some people with numerous intestinal parasites. The frequency of protozoan infections (13.9%) was higher than the helminthic infections (2.6%). Blastocystis spp. (8.1%) was the most prevalent detected intestinal protozoan. Entamoeba coli (5.7%), Dicrocoelium dendriticum (2.2%), Giardia lamblia (1.5%), Fasciola spp. (0.2%), and Hymenolepis nana (0.2%) were other detected parasites.

In- spite of betterment of the health condition in Iran and reduction of parasitic infection, intestinal parasitic infections are still a considerable public health issue in some parts of Iran.

Echinococcosis is the most common cestode infection globally caused by the Echinococcus species. The most common organ involvement is the lungs and liver, but other organs can also be rarely involved. Here, we present a case with a giant cerebral hydatid cyst.

Case report: 

A 4-year-old boy presented with abnormal gait and walking. Cranial magnetic resonance imaging showed a cyst of 13 cm in diameter. The cyst was enucleated successfully with no rupture. Oral albendazole therapy was started. There was no eosinophilia, and the echinococcal indirect hemagglutination test was negative. Ultrasonography detected an anechoic cystic lesion in the liver. He was evaluated for deep-organ involvement, however, no cysts were detected in other organs. The histopathological examination was compatible with a hydatid cyst.

Although intracranial hydatid disease in children is rare, it should be considered among the differential diagnoses in patients with neurological symptoms, especially in highly endemic regions.

Schistosomiasis is a parasitic disease caused by trematodes (body flukes), affecting millions worldwide. However, its pulmonary manifestations are rare. We report a rare case of a 51-year-old People Living with HIV male, managed in a tertiary care hospital in west India in May 2023, vegetable vendor who was admitted with complaints of dysphagia, odynophagia, fever and chest pain for 3 days, cough and breathlessness for 1 month. Chest x-ray and CT scan were suggestive of hypodense fluid collection with rim enhancement along right lateral and posterior aspect of thoracic esophagus. All routine investigations and urine cultures were sent, which turned to be inconclusive. Upper Gastrointestinal scopy was suggestive of pangastritis. Fiberoptic bronchoscopy was done with no structural abnormality or endobronchial mass. Bronchoalveolar lavage from right lower lobe was sent for CBNAAT, Gram and Ziehl Nelson staining and cultures, acid fast bacilli cultures and cytology which revealed parasitic infection with Schistosoma haematobium. The patient was treated with tablet praziquantel P/O 2400 mg in divided doses for 1 day followed up after two weeks when he experienced reduced symptoms. Sputum examination was repeated showed Schistosoma on wet mount and hence a repeat dose of tablet praziquantel 3000 mg in divided doses was given and was advised to follow up 2 weeks later, which showed resolution of right lower zone opacities.

Hydatid cyst is an infestation caused by Echinococcus granulosus, which is endemic in most countries and most commonly affects the liver and lungs. Isolated muscle involvement is very rare and accounts for 3-5% of all cases. In April 2022, Health Sciences University Elaziğ Fethi Sekin City and Research Hospital in Turkey received a referral for a 37-year-old male patient from the physical therapy and rehabilitation outpatient clinic. The patient presented with complaints of chronic pain in his left shoulder and back, which worsened with movement. After the radiological and serological confirmation of diagnosis, the patient underwent wide surgical resection, adjuvant and neoadjuvant anthelmintic chemotherapy. Precise and complete removal of the cyst without infecting the surrounding tissues combined with anthelmintic chemotherapy will not only increase the success of the surgery but will also minimize the possibility of recurrence and distant organ spread. In this study, we aimed to raise awareness by presenting the first reported isolated hydatid cyst in the infraspinatus muscle case from our country, which is extremely rare.