Iranian Journal of Parasitology
Volume:16 Issue: 1, Jan-Mar 2021

Iran is one of the endemic areas of Mediterranean Visceral Leishmaniasis, a disease caused by Leishmania infantum. In this work, we examined whether Proteína quimérica 10 (PQ10) recombinant protein is suitable for immunological diagnosis of human visceral leishmaniasis.

The study was carried out in Tarbiat Modares University during 2016- 2018. The coding sequence of PQ10 recombinant protein was sub-cloned in pET28 expression vector and was commercially synthesized by GENERAY Biotechnology, China. Sequencing with proper primers was done, the expression, optimization of expression and protein purification were performed, and the purified recombinant protein was confirmed by western blot. The efficacy of PQ10 for serodiagnosis was evaluated with 50 positive and 50 negative serum samples, which confirmed by the direct agglutination test and collected from individuals living in the visceral leishmaniasis endemic areas of Iran. ELISA was performed with the PQ10 recombinant protein.

The 95% CI sensitivity of ELISA that was evaluated with sera from naturally infected individuals was 84%. The 95% CI specificity value of the ELISA determined with sera from healthy individuals (50 serum samples) and from individuals with other infectious diseases was 82%. The 95% CI positive predictive value (PPV) and negative predictive value (NPV) were exterminated 82.35% and 83.67%, respectively.

We have used a recombinant synthetic protein to improve serodiagnosis of human visceral leishmaniasis. PQ10 could be useful for diagnosis of asymptomatic cases, as well as in the early phase of infections.

Cystic echinococcosis (CE) is one of the most important parasitic zoonosis in the world. Post-surgery follow up in CE patients is an important non-solved problem up to now. Therefore, the investigations on this problematic issue would be very applicable in the view of CE clinical treatment.

A total of 24 confirmed liver CE patients sera including eight sera before surgery (BS), eight sera three months post-surgery (3MPS), and eight sera six months postsurgery (6MPS) were used in the present study. Proteomics methods including 2DE and LC-MS/MS were performed on the specimens followed by bioinformatics analysis such as Gene Ontology (GO) and Protein-Protein Interaction (PPI) network analysis.

A total of 235 proteins were detected of which 12 differentially expressed proteins (DEP) were identified by LC-MS/MS in all sera. The proteins were presented in BS and suppressed after surgery as follows: HPX, SERPINA1, SERPINC1, CP, HBD, and HBA2. Comparisons of the protein expression in sera of patients BS, 3MPS, and 6MPS revealed that GC, IGJ, AHSG, CD5L, FGG, and APOC3 have been overexpressed in 3MPS and 6MPS. PPI network analysis demonstrated that SERPINC1 and AHSG with more connection in the network could be considered as hub proteins and potential prognostic biomarkers in response to surgical treatment of liver CE.

Application of proteomics methods on patient’s sera could be used as a novel biomarker tool for following-up liver CE patients. In this regards, proteomics and, application of bioinformatics analysis including GO and PPI showed that SERPINC1, AHSG and HPX are of more value as a potential follow up biomarkers in response to surgical treatment.

Toxocara cati is considered as one of the main etiological agents of toxocariasis with global and regional importance. As there is no information on proteomics of T. cati, herein, we reported the results obtained by proteomic analysis of somatic proteins extract, using a mass spectrometry (LC–MS/MS) approach.

Somatic extract fractions were separated by two-dimensional SDSPAGE and were electro blotted on to PVDF membranes for immunoblot analysis, then collected the immunogenic spots which response of antibodies of the paratenic hosts (mice) to the antigens ( Mashhad, 2017), and analyzed by LC–MS/MS. The LC-MS/MS data were analyzed by Mascot database, Taxonomy Toxocara, and common contaminants, in Omics Center, Biotechnology Medical University of Graz (Austria, 2018).

The protein spots were isolated between 15–140 kDa ranges using 3–10 non-linear IPG strips and Brilliant Blue Coomassie. Ten proteins were characterized as immunogenic proteins, seven of them were identified and three of them were unknown proteins.

This study provided additional information about the somatic antigens of T. cati, which can lead to the development of new strategies for novel immuno-modulators, drug targets, subunit vaccines and immunodiagnostic kits for toxocariasis.

The present study aimed to assess the therapeutic effect of chitosan nanoparticles and metronidazole against Giardia lamblia as well as evaluate the efficacy of loading metronidazole on chitosan nanoparticles.

This study was carried out at medical Parasitology Department, Faculty of Medicine, Zagazig University and Theodor Bilharz Research institute (TBRI) from February 2019 to February 2020 on 45 hamsters. They were divided into 5 groups 9 hamsters each: Group A non-infected hamsters, Group B infected control group, Group C, D and E infected with G. lamblia and treated with Chitosan nanoparticles (CsNPs), metronidazole (MTZ) and metronidazoleloaded chitosan nanoparticles (MTZ-CsNPs) respectively.

The highest percentage of reduction in the Giardia cyst and trophozoite counts were in group that received MTZ-CsNPs (94.69%, 94.29%). Lower percentages of reduction were recorded for MTZ treated group (90.15%, 89.52%) and CsNPs treated group (63.64%, 75.24%). Histopathological examination showed marked healing of intestinal mucosa after treatment with MTZCsNPs.

CsNPs showed a therapeutic effect against Giardia infection in hamsters. Loading of metronidazole on chitosan nanoparticles enhanced therapeutic effect of both CsNPs as well as metronidazole.

Placental malaria has ability to upregulate prostaglandin synthesis by increasing cyclooxygenase-2 (Cox-2) enzyme activity. Cox-2 and prostaglandin have a role in causing uterine contraction and therefore can cause abortion or preterm labor. Tablet AS201-01 containing the ethyl acetate fraction of Andrographis paniculata was tested in vivo on pregnant mice infected with Plasmodium berghei. AS201-01 inhibited the growth of P. berghei, increased TGF-β expression, decreased TLR-4 expression and apoptosis index of placental tissue in P. berghei infected pregnant mice and thus prevented placental malaria complications. These effects were correlated with the decrease of Cox-2 and prostaglandin expression.

Twenty-four pregnant mice (Balb/c) were divided into 4 groups (n=6). Mice were maintained at Animal Laboratory of Institute of Tropical Disease, Universitas Airlangga, Surabaya, Indonesia in 2016. G1 were uninfected pregnant mice, G2 untreated infected pregnant mice, G3 infected pregnant mice treated with AS201-01, and G4 infected pregnant mice treated with DHP tablet. All infection groups (G2-G4) were inoculated with 1x106 of P. berghei parasite on day 9 of gestation and treated on day 11. All mice were terminated at day 15 of gestation, and placental tissue was collected. Cytokine expression of Cox-2 and prostaglandin were evaluated using immunohistochemistry.

G3 was found to have lower Cox-2 and prostaglandin expression compared to G4 and G2, but higher compared to G1. Cox-2 and prostaglandin expression was significantly different among groups (P<0.001).

This study demonstrates the ability AS201-01 tablets have to decrease Cox-2 and prostaglandin expression on placental of P. berghei infected mice and therefore eliminates the adverse effects of placental malaria.

Snails of the genus Galba are the intermediate hosts of Fasciola species, the etiological agents of liver fluke disease, fascioliasis. A genetically different but morphologically very similar species in the genus, G. schirazensis, is sympatrically distributed with G. truncatula in some regions of the world. We aimed to investigate the occurrence of G. schirazensis in Kerman province, Iran and to characterize genetically G. schirazensis specimens from southeast Iran.

Field-collected snails from four localities in Jiroft, Bam and Faryab, Kerman province, southeastern Iran were studied. Hydrological variables including temperature and pH were recorded for each habitat. Each specimen was identified using morphological as well as conchological characteristics. Genetic characterization was performed using PCR-sequencing followed by phylogenetic analyses on nuclear ITS2 as well as mitochondrial cox1 gene fragments. MaxEnt software was used to predict the most appropriate ecological niches for the targeted species.

G. schirazensis was found in 4 out of 28 locations. One ITS2 and two cox1 haplotypes were detected among G. schirazensis populations from the four localities. Habitat study showed that G. schirazensis thrives in habitats with alkaline pH. G.schirazensis from South America were clustered with specimens from Bam, Kerman, Iran; however, north Iranian isolates of G. schirazensis were strongly correlated with specimens from Jiroft and Faryab. MaxEnt model for the most appropriate ecological niches of the targeted species predicted environmental suitability for this species in western Africa as well as coastal areas in north and southwestern Africa.

G. schirazensis is frequently present in southern areas of Kerman Province. At least two genetically different haplotypes are present in southeastern Iran.

The present study was conducted in Jul 2019 and Jan 2020 in two wildlife parks of the Nowshera district, Khyber Pakhtunkhwa, Pakistan, where the endangered Punjab urial (Ovis vignei punjabiensis) is successfully bred in captivity. We determined diversity of internal and external parasites that take advantage of the situation of congestion, resulting in massive mortalities of wild animals in captivity.

Internal parasites of living urial were determined by direct wet smear and flotation methods, while dead urial was necropsied for any pertaining observation.

All examined fecal samples were found infected with gastrointestinal parasites, which had significant difference in the total abundance in winter and summer. S. papillosus and H.contortus, and a single protozoan, Eimeria spp. were the dominant parasites in fecal samples. Ticks collected from urial enclosures and dead animals were of single species H. anatolicum. Theileria spp. was observed in blood, while hydatid cysts were found in lungs and liver of necropsied urial.

The study indicates that internal parasites such as Haemonchus contortus and Strongyloides papillosus, while external parasites as Hyalomma anatolicum ticks played major role in the population decline. Strict veterinary control of infectious diseases, provision of hygienic and supplementary diet, and proper maintenance of urial population are necessary measures for the control of mortalities.

Human infection with Strongyloides stercoralis and hookworm parasites is usually under reported due to less sensitive diagnostic methods. Agar plate culture (APC) is the most sensitive technique for parasites having larval stage. However, using APC in routine diagnosis is uncommon. This study aimed to determine the detection rate and sensitivity of APC in comparison with formal ether concentration technique (FECT) and spontaneous tube sedimentation techniques (STSTs) for S. stercoralis and hookworm larvae.

Stool samples collected from 844 schoolchildren in Amhara Regional State, northwestern Ethiopia in 2019, transported to nearby health institutions and processed by APC, FECT and STSTs. The prevalence of S. stercoralis and hookworm was computed by descriptive statistics and Chi-square. The diagnostic agreement among the three techniques was evaluated using Kappa value.

The overall prevalence of S. stercoralis and hookworm infections by combining the three methods was 13.2% (111/844) and 33.8% (277/844), respectively. Using APC alone, the prevalence of S. stercoralis and hookworm were found to be 10.9% (92/844) and 24.5% (207/844), respectively. Agar plate culture was 5.4 and 2.7 times respectively more sensitive than FECT and STST, with slight and fair agreement in the detection of S. stercoralis. Hookworm diagnostic agreement was moderate between APC and FECT, and APC and STST. The Kappa value between STST and FECT diagnostic methods was substantial.

APC has a better detection rate of S stercoralis and hookworm larvae. Therefore, APC can be used as an alternative routine diagnostic method to S. stercoralis and hookworm co-endemic countries.

The present study aimed to determine genetic diversity of Trichomonas vaginalis (T. vaginalis) isolates with microsatellite markers in Turkey (Nov 2015 to 2016) and to create a web-based microsatellite typing (MT) approach for the global interpretation of the data. In addition, the endosymbiosis of Mycoplasma hominis (M. hominis) and T. vaginalis virus (TVV) in the isolates was also examined.

The allele sizes for each locus were calculated and microsatellite types were determined according to the allele profiles. The population structure was examined with Bayesian clustering method. A website (http://mttype.adu.edu.tr) was created for collection and sharing of microsatellite data. Presence of TVV and M. hominis in T. vaginalis isolates were investigated with electrophoresis and PCR.

Of 630 vaginal samples T. vaginalis was detected in 30 (4.7%) and those were used for further analysis. The structure produced by a clustering algorithm revealed eight genetic groups. The typing of isolates according to microsatellites revealed 23 different microsatellite types. Three clones were determined among isolates (MT10 16.7%; MT18 10% and MT3 6.7%). The frequency of TVV and M. hominis was 16.6% (n=5) and 20% (n=6), respectively.

Presence of three clones among 30 T. vaginalis isolates indicated that microsatellite-based genotyping was efficient to determine the clonal distribution of T. vaginalis isolates. Therefore, a promising tool might be developed further and adapted to the studies dealing with molecular epidemiology of T. vaginalis. Microsatellite data from forthcoming studies will be deposited and presented on the website. In addition, we also presented the frequency of two endosymbionts in T. vaginalis isolates for the first time in Turkey.

Schistosomiasis has been identified as a major public health problem in tropical countries. The present study aimed to investigate the schistosomicidal effects of the methanolic extract of Argemone mexicana L. and its active component, berberine against Schistosoma mansoni on in-vitro experiments.

S. mansoni adults were used. Various concentrations of the methanolic extract (10 - 200 µg/ml) and berberine (2.5 - 50 µM) were tested from 24 to 72 h. The viability of S. mansoni was confirmed with an invertoscope-microscope. Furthermore, cytotoxic (Hemolysis test), and antioxidant (DPPH radical scavenging assay) capacities were determined.

The viability tests on S. mansoni showed that A. mexicana at 50 μg/mL is lethal at 48 h and berberine at 10 μM is lethal at 24 h. The hemolytic activity at 1,000 μg/mL was 2.9% for A. mexicana and 90.2% for berberine. The antioxidant capacities shown by A. mexicana and berberine, were EC50 156.3 and 84.1 μg/mL, respectively.

The extract of A. mexicana and berberine demonstrated high antischistosomal activities in low concentration and short exposure time on the in-vitro model.

In this study, we assessed the in vitro antischistosomal activity of the active ingredients of Allium sativum (allicin) and Curcuma longa (curcumin) on Schistosoma mansoni.

This study was conducted in Faculty of Science, Port said University, Egypt (2018). Adult worms were exposed to a range of concentrations of AL or CU, and worm survival was assessed 24 h post-exposure to calculate the lethal concentration of the compounds. Scanning electron microscopy was used to assess ultrastructural changes in the surface of AL- or CU- treated worms. The genotoxicities of AL and CU on S. mansoni were determined by DNA fragmentation analysis.

We determined the concentrations of AL and CU required to kill 50% of S. mansoni (LC50). The LC50 of AL was 8.66 µL/mL, whereas 100% mortality of S. mansoni was achieved by AL at concentrations of 50 µL/mL. The LC50 of CU was 87.25 µL/mL, with the highest mortality of 91.3% seen after 24 h exposure to 100 µg/mL CU. Ultrastructural studies revealed that exposure to either AL or CU led to mild or severe surface damage to S. mansion, respectively. The degree of damage in the worms was sex-dependent. Interestingly, while CU exposure resulted in DNA fragmentation in S. mansoni worms, we observed no genotoxic effects of AL.

Both AL and CU exhibit antischistosomal activity; the study provided evidence suggesting that these compounds act through distinct mechanisms. These promising results encourage further investigation into these compounds as potential antischistosomal agents, either alone or as complementary treatments to praziquantel.

The genus Acanthamoeba is a free-living opportunistic protozoan parasite, which widely distributed in soil and fresh water. Acanthamoeba keratitis, which causes a sight-threating infection of the cornea, is going to rise in Iran and worldwide. The aim of this study was to compare direct microscopy, culture and PCR for detection of Acanthamoeba spp. in clinical samples and to determine the genotypes of Acanthamoeba spp. by sequencing 18SrRNA gene.

Among patients clinically suspected to AK referred to a tertiary ophthalmology center at Mashhad, northeastern Iran. During 2017-18, twenty corneal scrapes specimens obtained. The samples were divided into three parts, subjected to direct microscopic examination, culture onto non-nutrient agar and PCR technique. Sensitivity, specificity, accuracy and likelihood ratio were evaluated.

Among 20 persons clinically suspected to amoebic keratitis, 13(69.2%) patients definitely diagnosed as Acanthamoeba keratitis. Wearing contact lens, eye trauma due to foreign particle and swimming in fresh water were the main predisposing factors. Most of patients suffered from pain and photophobia. Corneal ring infiltration and epithelial defect were common clinical sings. Direct examination had the lowest sensitivity and sensitivity of both Nelson-PCR and JDP-PCR methods were equal and highest. In addition, the results of sequencing identified that all strains belonged to T4 genotype.

Amoebic keratitis is a sporadic parasitic keratitis, which is mainly seen in contact lens user in Mashhad. PCR based on 18S ribosomal DNA with JDP primers is a reliable and highly sensitive method for diagnosis of Acanthamoeba keratitis in clinically suspected cases

Trichinellosis is a serious food-borne parasitic zoonosis, thus finding high quality antigens is the key to serodiagnosis of trichinosis. This article reports the characterization and sensitivity of four recombinant proteins expressed by four genes (Wn10, Zh68, T668, and Wm5) from different developmental stages of Trichinella spiralis for the diagnosis of trichinellosis in mice.

This study was conducted in Jilin University and National Institute of Parasitic Diseases of Chinese Center for Disease Control and Prevention in 2017-2018. The structures and functions of the proteins encoded by four genes were predicted by bioinformatics analysis. The four genes were cloned and expressed, and the recombinant proteins were purified. Anti-Trichinella IgM and IgG antibodies in the sera of mice infected with T. spiralis from 1-45 d post-infection (dpi) were evaluated by ELISA.

The optimal antigen epitopes of four proteins (P1, P2, P3, and P4) encoded by the four genes from T- and B-cells were predicted, and four purified recombinant proteins (r-P1, r-P2, r-P3, and r-P4) were successfully produced. For IgM, the antibody levels detected by the four recombinant antigens were approximately equal to the cut-off value. Anti-Trichinella IgG antibodies were first detected by r-P1 at 8 dpi, followed by r-P2, r-P3, and r-P4 at 10 dpi, 14 dpi, and 16 dpi, respectively, and the antibody levels remained high until 45 dpi.

The recombinant antigens r-P1, r-P2, r-P3, and r-P4 could be antigens that react with antibodies, they showed high sensitivity in the detection of anti-Trichinella IgG antibodies in mice. Among these proteins, r-P1 may be a candidate antigen for the detection of anti-Trichinella IgG antibodies in the early infection phase and exhibited the best sensitivity among the antigens.

The present study aimed to control mebendazole drug release from ethyl cellulose nanofibers containing guar gum produced by Electrospinning Method (ESM) on mortality of hydatid cyst protoscoleces under laboratory conditions.

The study was conducted in Arak Islamic Azad University, 2019. After preparation of ethyl cellulose nanofibers containing guar gum with concentrations 10, 250, 50 and 500 ppm with ESM, the uniformity and fineness of nanofibers were investigated by electron microscope. By determining the absorption of nanofibers during 312 h via spectrophotometry method, the amount of drug release was obtained. Then, the mortality of live protoscoleces in-vitro with nanofibers made with different concentrations was studied during 13 days.

Guar gum nanofiber with four concentrations of 10, 50, 250 and 500 ppm had 0.78512, 0.83729, 1.0098 and 1.0633 absorption respectively and showed drug release 42.09%, 39.95%, 33.05% and 30.96% after 312 hours. Therefore, the survival of protoscoleces in the presence of guar gum with four concentrations was zero after 3, 6, 11 and 13 days (P<0.05).

To produce nanofibers carrying the drug for research related to the treatment of hydatid cysts, the electrospinning technique can be considered as a reliable method.

Dogs, as the definitive host of Neospora caninum, are important in the epidemiology of this parasitic infection. We aimed to determine the prevalence of N. caninum infection in a dog population from a rural setting in Fars Province, Southern Iran, using a combination of molecular and serological techniques.

This cross-sectional study was carried out in Nov 2018 in three rural districts, Sar Mashhad, HosseinAbad, and Tolesaman located in Kazeroun Township in Fars province, southern Iran. Blood samples were taken from 60 stray and household dogs. Dogs’ sera were tested for antibodies against N. caninum, using a Neospora-Modified Agglutination Test. Moreover, dogs’ buffy coats were tested for Neospora DNA, using a molecular method.

Anti-Neospora antibodies were detected in sera of 4 out of 60 dogs, corresponding to a seroprevalence rate of 6.7%. Out of 25 female dogs, 1 was seropositive and of 35 males, 3 were seropositive, yet the differences were not statistically significant. The infection was more prevalent in adult dogs (> 12 months), nevertheless, the differences between age and Neospora seropositivity was not statistically significant. N. caninum DNA was not detected in the buffy coat of any of the studied dogs.

Findings of the study indicate that N. caninum is a common infection in dogs in rural areas of Fars province in southern Iran. The infected dogs might be a potentially important source of N. caninum infection to livestock in the area.

Due to numerous side effects of common drugs in treatment of leishmaniasis, new therapeutic approaches focus on herbal compounds. Therefore, we aimed to determine the effect of crocin and stigmasterol on in-vitro growth of promastigotes and amastigotes of Leishmania major in the Department of Parasitology, Pasteur Institute, Tehran, Iran in 2018.

The effect of different concentrations of crocin and stigmasterol were evaluated by determining their in-vitro inhibitory effects on promastigotes and amastigotes of the L. major using MTT assay.

The fatality rate was 65.27% and 71.96% for crocin and stigmasterol respectively at 24 h post-culture in concentration of 50 μg/mL. The mean inhibitory effect of crocin and stigmasterol on L major amastigotes after 72 h were 52.22% and 38.96%.

The crocin and stigmasterol had efficient adverse effects on promastigote and amastigotes of L. major, hence, further studies on the anti-leishmanial effects of these herbal compounds in human and animal models are recommended.

Chronic granulomatous disease (CGD) described as an essential immunodeficiency problem of phagocytic cells resulting in a phagocyte dysfunction and inability to kill a spectrum of bacteria and fungi. Despite the fact that CGD patients are more susceptible to intracellular infections, visceral leishmaniasis has been reported rarely in these cases. Here, we report an uncommon case of visceral leishmaniasis in a child with CGD. An 8-yr old boy with CGD presented to the infectious disease ward, Children's Medical Center, Tehran University of Medical Sciences, Iran after the onset of 20 days fever with chronic crusted ulcer approximately 3 cm × 3cm on the left upper limb and a small ulcer measuring 0.5 cm × 0.5 cm on the right knee with moderate secretion. Bone Marrow Aspiration (BMA) and Bone Marrow Biopsies (BMB) of fragmented samples were performed and polymorphic population of hematopoietic cells, Megakaryocytes and Leishman bodies were seen. The patient was treated with meglumine antimoniate (Glucantime®) 20 mg/kg for 28 days and after partial improvement patient discharged and continue the treatment at home. Amphotericin B lipid complex (Ambisome®) (3–5 mg/kg per dose once) was administered every 3 -4 weeks for 18 months as secondary prophylaxis that was well tolerated and effective.

Hydatid disease is a parasitic infection caused by Echinococcus granulosus with worldwide distribution. The most affected organs are liver and lungs, but it can be detected in any other organs as well. We reported a 5-yr-old boy from Shiraz, southern Iran in 2017 who presented with abdominal discomfort. Imaging revealed multiple liver hydatid cyst and a huge kidney hydatid cyst. This case showed the possible implication of rapid growth of multiple hydatid cyst as well as unusual organ presentation in the pediatric population.

Morbidity of mixed cystic and alveolar echinococcosis (CE and AE) is exceptionally rare. Less literature retrieved from a database on the internet detailed the content, including radiography, pathology, and therapeutics data. Such a case of co-occurrence of the different Echinococcus species was diagnosed and treated at our hospital center from Nov 2019 to Feb 2020. A 30 yr old female from the pastoral area in Qinghai Province, China, was diagnosed with a case of echinococcosis and diagnosis was confirmed after image studies, immunoassaying of hydatid enzymes, life history and pathology result. The patient underwent hepatectomy along with excision of the internal capsule. Post-operative pathology was done, and it confirmed a mixed infection of both CE and AE. The patient recovered well without complications after liver-protecting and tissue repair treatment for 15 days. Knowing about infective mode and immune method of the case might be vital for research on variation for Echinococcus infection.