Background: Although rigorous efforts have substantially decreased the malaria burden through decades, it still threatens the lives of millions of children. Development of an effective vaccine can provide important approach in malaria control strategies. Unfortunately, development of an effective vaccine for falciparum malaria has been hindered by the extreme complexity of malaria parasite biology, complex and diverse parasite genomes, and immune evasion by the parasites as well as the intricate nature of the parasites infection cycle. The aim of this review was to discuss the different approaches to malaria vaccine development until now.

Methods: Scientific databases, including MEDLINE (via PubMed) and SCOPUS were searched up to 30 Jan 2017 and the articles regarding malaria vaccine development were taken into examination.

Results: Several strategies for malaria vaccine development including pre-erythrocytic vaccines, antibody-based subunit vaccines, vectored vaccines, whole sporozoite vaccines, genetically Attenuated parasites and sporozoite subunit vaccine, erythrocytic vaccines, sexual stage vaccine, transmission-blocking vaccine as well as synthetic peptides and conjugate vaccine has been introduced. However, the success has been limited thus far.

Conclusion: Although development of malaria vaccine over the past 70 year has been continued, the discovery, development, and licensing of a malaria vaccine formulation, which meets safety, affordability, accessibility, applicability, and efficacy has not yet been achieved.

Background: Currently, there is no vaccine available for any form of leishmaniasis for human use, including visceral leishmaniasis (VL). The treatment relies on drugs associated with severe toxic side effects and increased parasite drug resistance. At present, there is a strong need to develop and implement a successful vaccine against this disease. Therefore, we evaluated immunoprophylactic potential of a cocktail of low molecular weight antigens along with various adjuvants.

Methods: The three antigens (2015, Department of Zoology, Panjab University, Chandigarh), 31kDa, 36 kDa and 51 kDa of L. donovani were used in this study. Inbred BALB/c mice were immunized with 10 µg of cocktail antigens i.e. 31+36+51kDa alone and along with different adjuvants (ALD, saponin, and liposome). Mice were boosted twice at an interval of 2 wk and after last dose; mice were given challenge infection with 10⁷ promastigotes. Mice have sacrificed15 d post immunization and on 30, 60, 90 post-challenge days for evaluation of different parameters.

Results: Immunized animals showed reduced parasite load, increased DTH responses and elevated levels of IgG2a antibody. The levels of Th1 cytokines were higher as compared to Th2 cytokines in immunized animals.

Conclusion: Best results were obtained with cocktail of 31+36+51+liposome and this combination conferred maximum protection.

Background: Diarrheal disease annually causes 760000 deaths in children, and 1700 million new cases are reported each year worldwide. Among the parasites, Entamoeba histolytica, Giardia intestinalis, and Cryptosporidium spp. are the most important infectious agents leading to diarrhea. Clinical presentations due to these parasites are more or less similar, and microscopy is not as much as sensitive for the detection. The aim of this study was to set up and evaluate a Multiplex PCR Assay for Synchronous Identification of Entamoeba histolytica, Giardia intestinalis, and Cryptosporidium spp. in Stool Samples

Methods: Samples were obtained from different sources such as culture media and patient stool samples. Primer pairs were designed using primer-BLAST, and for the extraction of DNA, the QIAamp DNA stool mini kit was used. The study was conducted in Tehran, Iran and completed in 2016.

Results: The current multiplex PCR assay for the detection of E. histolytica achieved sensitivity and specificity of 86.36% (95% CI: 65.09% to 97.09) and 95.74 % (95% CI: 85.46% to 99.48%), respectively. Sensitivity and specificity of the test for G. intestinalis was 90.91% (95% CI: 70.84% to 98.88%) and 95.74% (95%CI: 85.46% to 99.48%), respectively, and for the detection of Cryptosporidium, multiplex PCR showed a sensitivity of 90.91% (95% CI: 70.84% to 98.88%) and specificity of 95.74% (95%CI: 85.46% to 99.48%).

Conclusion: Multiplex PCR in this study showed admissible sensitivity and specificity for the detection of E. histolytica, G. intestinalis, and Cryptosporidium spp. in fecal samples.

Background: For many years, malaria was a major life-threatening parasitic infection in Iran. Although malaria elimination program is implementing in the country, still some cases annually are reported from malaria-endemic areas.

Methods: This study was conducted in five malaria endemic districts of Sistan and Baluchistan Province, southeastern Iran, neighboring Afghanistan and Pakistan countries. Overall, 170 and 38 vivax malaria and falciparum malaria infected patients were enrolled in the study from 2013-2014. All the cases were selected according to criteria of the WHO guideline for in vivo drug sensitivity tests in malaria parasites. Evaluation of drug sensitivity test was conducted with some modifications.

Results: The patients with vivax malaria responded to the regimen of chloroquine in 37.4(±15.9), 40(±13.8) and 42(±17.7) h for Pakistani, Iranian and Afghani nationalities respectively based on MPCT evaluation. The results showed a considerable difference between them and Iranian subjects. MPCT for the patients with falciparum malaria was calculated as 28(±18.05), 26(±12.03) and 36(±16.9) h for Iranian, Pakistani and Afghani nationalities respectively. There was a marginally significant difference between Afghani and other nationalities and between males and females.

Conclusion: Treatment of all the patients resulted in ACPR and MPCT of P. vivax showed that the parasite became more sensitive to chloroquine than previous years in studied areas.

Background: Toxoplasmosis is a parasitic disease caused by the intracellular protozoan parasite, Toxoplasma gondii, which can infect humans and warm-blooded animals. This infection can lead to still birth and abortion among some susceptible hosts especially sheep and human in pregnancy. Development of a vaccine against T. gondii infection is very important-especially for use in immunocompromised patients, pregnant women, and sheep. Different antigens of T. gondii can be potential candidates for immunization. The aims of this study were to identify the immunodominant and antigenic proteins of T. gondii in sheep and man.

Methods: Tachyzoites’ proteins were separated by two-dimensional polyacrylamide gel electrophoresis (2-DE), and subjected to western blot analysis probed with T. gondii positive sera of sheep and human (Biotechnology Department of Pasteur Institute of Tehran, Iran, from April 2016 to March 2017). Finally, the immunoreactive proteins were identified by mass spectrometry (MALDI-TOF/MS and MS/MS) technique.

Results: Five immunoreactive and antigenic proteins were recognized by Toxoplasma positive sera of human and sheep. These identified proteins were Enolase 2, rhoptry protein 4 (ROP4), dense granular protein 14 (GRA14), rhoptry protein 15 (ROP15) and rhoptry protein 9 (ROP9).

Conclusion: The identified immunodominant proteins have potential to be used as diagnostic antigens and as diagnostic markers of Toxoplasma infection in sheep and human.

Background: C type lectin (CTL) family is a type of calcium-dependent proteins found in vertebrates and invertebrates. The objective of this study was to perform a comparative analysis and phylogenetic inferring for understanding the similarities and differences of carbohydrate recognition domain (CRD) domain of Toxocara canis CTL and other nematodes, and similar C type lectin involved in the immune system of mouse and human as their host.

Methods: The female T. canis was retrieved from the 2-6 months puppies (Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran, 2015). To collect T. canis eggs, the worms were cultured for 5 d until they were embryonated. The hatching process was accelerated for collecting the stage 2 larvae, and the larvae were cultured for a week. A cDNA library was made from the total mRNA of T. canis infective larvae. The PCR amplification for C type lectin gene was performed and the amino acids were analyzed using the alignment method and the construction of phylogenetic tree.

Results: The suspension sample maintained at 30 ºC for four weeks could embryonate 90%-100% of eggs. T. canis CTL gene was 657 bp in length and encoded a protein with 219 amino acids. The CTL of species of Strongylida order were closely placed in the tree, whereas the members of Ascaridida orders were located in a separate branch. High levels of similarity (36%-44%) and conservation of C type lectin from T. canis with mouse and human C type lectins. Its C type lectin showed a higher similarity with asialoglycoprotein receptor (ASGPR), macrophage lectin, dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN), MINCLE receptor of mouse and human.

Conclusion: Analysis of CRD domain of C type lectin protein could make a better understanding of their role in the interaction of nematode parasite with their hosts.

Background: We aimed to determine the incidence of trichomoniasis and its risk factors in Egyptian pregnant women attending the Minia Maternity and Pediatric University Hospital, Minia, Egypt and evaluate its association with preterm birth.

Methods: The study was carried out from Aug 2014 to Jun 2015 through 2 phases, the first phase was case-control study, and the second phase was follow-up with intervention. Overall, 300 pregnant women with gestational age of 20-36 weeks with no medical risk factors of preterm labour birth were enrolled. Vaginal swabs were examined by the wet mount microscopy and culture while urine samples were examined by urine analysis. Demographic information was collected. Pregnant women were divided into two groups, study group (with trichomoniasis) and control group (without trichomoniasis). Positive cases were subjected to metronidazole treatment.

Results: Thirty-five cases were positive for T. vaginalis infection. Maximum cases were detected by culture (11.7%) followed by wet mount microscopy (9.7%) whereas least number of cases (7.3%) was detected by urine examination. Nineteen (54.28%) cases had preterm delivery. Post-delivery adverse outcomes were observed in 29 cases (82.8%). The high rate of infection was observed in age group of 20-30 years (P<0.05). In addition, there was a signifi­cant T. vaginalis infection in pregnant women living in rural area, of low socioeconomic and primary educa­tional levels (P<0.05).

Background: Domestic dogs have been implicated as the main reservoir host of Mediterranean type of visceral leishmaniasis (VL) that is endemic in some parts of Iran. This study was performed about role of dogs in canine VL (CVL) epidemiology in Jiroft District, south of Kerman Province, southeastern Iran.

Methods: Totally, 165 dogs including 100 stray and 65 sheepdogs were randomly selected. After complete clinical examination blood sample was taken from each dog. All the collected samples were examined following the serum separation by direct agglutination test (DAT) for detection of anti-Leishmania infantum antibodies. The titers of ≥1:320 were defined as positive.

Results: Overall, of 165 serum samples, 13 samples (7.9%) were positive by DAT at titers of ≥1:320. The seroprevalence was 11% among the stray dogs and 3% among the sheepdogs. There was no significant difference between stray and sheepdogs in CVL infection. The highest seroprevalence rate (14.3%) was found in seven-year old dogs.

Background: The protozoan Trichomonas vaginalis is a sexually transmitted disease (STD). Metronidazole is a chosen drug for the treatment. This study evaluated the anti trichomonal activity of alcoholic extracts of combination Verbascum thapsus and Ginger officinale.

Methods: This experimental study was conducted in the Parasitology Laboratory, Kashan University of Medical Sciences, Kashan, Iran in 2015, on 23 women with suspected trichomoniasis referring to Kashan clinical centers. Medium TYI-S-33 was used for culture of three T. vaginalis isolates. Different concentrations (25, 50, 100, 200, 400, 800 µg/ml) of V. thapsus and G. officinale ethanol extract added to Trichomonas trophozoites in 48-well plates and metronidazole considered as positive control and the negative control was TYI-S33 containing Trichomonas trophozoites without any drug. In all of mentioned groups, trophozoites number counted 12, 24, 48 h after culture. Results were analyzed using ANOVA statistical test, to evaluate the toxicity of extract, measured by MTT assay. Induced apoptosis of T. vaginalis after treatment with different concentrations of extract was determined by Flow Cytometry.

Results: IC50 of alcoholic extract of combination V. thapsus and G. officinale and metronidazole after 24h was 73.80 µg/ml and 0.0326 µg/ml, respectively. The toxicity percentage of 25-800 μg/ml concentrations of this combination were between 0.2-1.98. In different concentrations of extract (25,50,100,200 and 400 µg/ml) apoptosis percent after 48h was 18.97 to 77.19 and necrosis percent was calculated 1.35, 3.18, 3.10, 1.16 and 4.09, respectively.

Background: There is paucity of information on functional relationship and characterization of Biomphalaria glabrata thioester-containing proteins (BgTEP) to other well-annotated homologues. We performed functional characterization studies of BgTEP to homologues in Anopheles gambiae and in disparate invertebrates.

Methods: Genomic sequences of TEPs were retrieved after annotation of the B. glabrata genome. In addition, TEP sequences deposited in NCBI protein database were also retrieved and utilized for sequence analysis. BgTEP relatedness to its other homologues as well as functional domain and protein-protein interaction analysis was performed.

Results: Our analysis resulted in the identification of TEPs in a number of organisms including, B. glabrata, A. gambiae, and Chlamys farreri. In addition, we identified 19 TEP sequences spread across 10 animal species. The B. glabrata genome contains 14190 unannotated proteins after filtration with about 85% of its proteome annotated. The phylogenetics, functional domain and protein-protein interaction analyses suggest an immunological role for BgTEP in B. glabrata.

Background: Due to the limited number of the available drugs for the treatment of trypanosomiasis, this study was designed to evaluate the trypanocidal effects of cisplatin or/and Nigella sativa oil (NSO) in experimentally infected mice with T. evansi.

Methods: During 2015 at the Parasitology Department, Menoufia University, Menoufia, Egypt, sixty Swiss albino mice were divided into six groups: normal control (I), infected control (II); cisplatin-treated (III); NSO-treated (IV); combined cisplatin + NSO-treated (V) and diminazene-treated (VI). The tested drugs were evaluated by the assessment of parasitaemia, measurement of aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, creatinine, serum IgM and a histopathological study.

Results: NSO showed a trypanocidal effect, however; it was not as effective as cisplatin or diminazene. There were significant increases of AST, ALT, urea, and creatinine in group II and III, which were significantly reduced in cisplatin + NSO-treated group (V). Moreover, there were significant reductions in serum IgM and the pathological changes of the examined organs of group V when they were compared with other treated groups.

Conclusion: Cisplatin combined with NSO showed a trypanocidal effect against T. evansi with preservation of vital organs functions and architecture.

Background: Irritable bowel syndrome (IBS) is a functional gastrointestinal disease with high population prevalence. Dientamoeba fragilis and Blastocystis hominis are reported worldwide as a cause of human gastrointestinal symptoms. This study evaluated the possible link between this syndrome and the infection with D. fragilis and B. hominis in Egypt.

Methods: Overall, 310 stool samples (160 from IBS patients and 150 from controls) were obtained from Tropical Medicine Outpatient Clinic, Faculty of Medicine, Ain Shams University, Cairo, Egypt between Mar 2015 and Feb 2016. All the stool specimens underwent direct examination and Trichrome staining. Each sample was cultivated on Jones and Robinson's media.

Results: Overall, 42 cases (28%) showed B. hominis and 2 cases (1.3%) for D. fragilis infections. After performing the culture methods for B. hominis and D. fragilis, detections increased to 50 cases (33.3%) and 3 cases (2%), respectively. While among 150 controls 18 (12%) positive samples were detected as B. hominis.

Conclusion: There may be a possible relationship between the presentation of irritable bowel syndrome and D. fragilis and B. hominis infections, which have to be excluded first.

Background: This study aimed to estimate the prevalence of trichomoniasis infection among females in Tehran, Iran.

Methods: This study was conducted on 482 women referred to the 6 obstetrics and gynecology centers of Tehran during 2015-2016. Some information including education, occupation, and number of sexual partners was obtained and clinical signs and symptoms of the genital tract were diagnosed by clinical examination. Two swabs were collected from the posterior fornix of patients. Two laboratory techniques, wet mount, and culture were carried out. Finally, statistical analysis test was performed using SPSS software version 16.0.

Results: Age distribution of patients was 15-60 yr. Trichomonas vaginalis was detected in 2 out of 482 participants (0.41%). All of the infected individuals were married (0.43%) and they had unique sexual partner and all of them had clinical symptoms. Significant association was observed between incidence of T. vaginalis infection and educational levels (P= 0.03), occupation (P=0.006), clinical symptoms (P=0.001), marriage (P=0.006) and bacterial infection (P=0.018).

Conclusion: The prevalence of trichomoniasis was low and its incidence was associated with several risk factors.

Background: Toxoplasma gondii is an obligate, intracellular parasite which causes the toxoplasmosis in humans and warm-blooded animals. Red meat is an important source for transmission of the infection to humans. This study aimed to determine the prevalence of Toxoplasma among imported and indigenous cattle in the Sistan region.

Methods: One hundred samples from slaughtered cattle were collected from two abattoirs of Zabol and Zahedan, South East of Iran in 2015. Each sample was a mixture of three muscle, including tongue, cardiac, and triceps. Additional data of each cattle, including sex, breed, age, indigenous or imported, location of slaughter, management practices, and feeding system were obtained through observations and interviews. Infection by T. gondii was determined by PCR method.

Results: The prevalence of Toxoplasma in indigenous cattle was 6% and in imported cattle was 26%, and this difference was statistically significant (P=0.006). Moreover, the prevalence of Toxoplasma was statistically associated with management practices (P=0.01) and feeding system (P=0.001). However, relationship between the prevalence of Toxoplasma with age, breed, sex, and location of slaughter was not statistically significant.

Conclusion: Since the prevalence of Toxoplasma among imported cattle is higher than indigenous cattle, so strict supervision for importing livestock from neighboring countries is necessary.

Background: Regarding hydatid cyst (cystic echinococcosis, CE) as a human public health problem in the West of Iran, molecular data related to the genotypes of Echinococcus granulosus in cattle and sheep in these regions are still insufficient. Here, we evaluated the genotypes of E. granulosus infecting sheep and cattle in western Iran.

Methods: Totally, 36 hydatid cysts including 18 hydatid cysts of sheep and 18 hydatid cysts of cattle were collected from Khorramabad slaughterhouse (Lorestan Province), Western Iran between May to September 2014. Protoscoleces or germinal layers were collected from individual cysts, DNA was extracted, and genotyping was performed by sequencing and analyzing mitochondrial cytochrome c oxidase subunit 1 (cox1) gene.

Results: In sequencing analysis, all of sheep isolates belonged to genotype G1 (sheep strain). Among cattle hydatid cyst isolates, 16/18 (88.9%) were belonged to genotype G1 and 2/18 (11.1%) were belonged to G3 genotype. The phylogenetic analysis showed two clusters; one of the clusters includes cattle G3 genotype and the other cluster represents sheep and cattle G1 genotype that were isolated.

Conclusion: The common sheep strain/G1 is predominant genotype in the western part of Iran, followed by G3 genotype, circulating among the animal hosts in this region. Further studies covering a larger number of isolates might be necessary to see if there are other genotypes in the hydatid cyst population in this region of Iran.

Background: This study evaluated the performance of routine permanent stain and cultivation method in comparison with polymerase chain reaction assay as the reference technique to detect Blastocystis sp.

Methods: A cross-sectional study was conducted among aboriginal populations that reside in Pahang, Peninsular Malaysia in Feb to Mar 2015. A total of 359 stool samples were examined using Wheatley’s trichrome stain, in-vitro cultivation in Jones’ medium and PCR assay. Positive amplicons were subjected to sequencing and phylogenetic analysis.

Results: Fifty-six (15.6%) samples were detected positive with Blastocystis sp. by Wheatley’s trichrome stain and 73 (20.3%) by in-vitro culture, while PCR assay detected 71 (19.8%) positive samples. Detection rate of Blastocystis sp. was highest in combination of microscopic techniques (27.9%). The sensitivity and specificity of Wheatley’s trichrome staining and in-vitro culture techniques compared to PCR assay were 49.3% (95% CI: 37.2-61.4) and 92.7% (95% CI: 89.1-95.4) and 39.4% (95% CI: 28.0-51.8) and 84.4% (95% CI: 79.7-88.4), respectively. However, the sensitivity [60.6% (95% CI: 48.3-71.9)] of the method increased when both microscopic techniques were performed together. False negative results produced by microscopic techniques were associated with subtype 3. The agreement between Wheatley’s trichrome stain, in-vitro culture and combination of microscopic techniques with PCR assay were statistically significant by Kappa statistics (Wheatley’s trichrome stain: K = 0.456, P<0.001; in-vitro culture: K = 0.236, P<0.001 and combination techniques: K = 0.353, P<0.001).

Background: Despite ducks being birds resistant to infection, the favorable habitat of ducks such as subtropical climate or stagnant water is also a perfect place for survival of the parasites.

Methods: This study was conducted from Dec 2014 to Apr 2015 to determine the prevalence of gastrointestinal parasites of domestic ducks in Ahvaz and environs, southwest of Iran. Overall, 41 fresh fecal samples were collected and prepared using formol-ether concentration, modified Ziehl-Neelsen, sheather`s floatation and zinc sulfate sedimentation methods. Light microscopic morphometry was used for identification of helminth eggs and oocysts.

Results: 60.97% of ducks were infected with three different nematodes and/or four protozoan parasites. The identified nematodes were Capillaria sp., (50%) Subulura spp. (16.66%) and Echinuria spp. (33.33%). The protozoan oocystes were Cryptosporidium spp. (50%) and coccidian species (%58.33) and included Wenionella philiplevinei, Tyzerria spp. and Isospora. mandari. Mixed infection with two or more parasites was common. Twenty (80%) had single, four (16%) double and one (4%) triple infection.

Conclusion: This is the first report of coccidian infection in domestic ducks of Iran. Further studies will be necessary on epidemiology and pathogenicity of the parasitic infections in ducks of this area.

Leishmaniasis, as a vector-borne disease, is transmitted by sandfly and caused by Leishmania protozoa. Brain involvement rarely occurs in visceral leishmaniasis. In this paper, a rare case of pons involvement by visceral leishmaniasis (VL) is reported. A 54 yr old man from Southwest of Iran (Yasuj) presented to the Emergency Ward with a 3-wk history of headache (continuous, throbbing, and general), fever, chills, weakness, anorexia, and weight loss.

Wild boars may be infected with several zoonotic parasitic infections including Fasciola spp. We reported a case of Fasciola infection in a wild boar in Bushehr Province in southwestern Iran. The sample was isolated from the liver of a hunted wild boar. A few of adult worms were fixed and stained. DNA was extracted from apical and lateral parts of the worms and PCR amplified, targeting NADH dehydrogenase subunit 1 (nad1) and cytochrome C oxidase subunit 1 (cox1) mitochondrion genes. Although the worm was quite long and looked much similar to F. gigantica, sequencing and analysis of PCR products of nad1 and cox1 genes revealed that the isolate has the most similarity with F. hepatica. This is the first report of molecular evaluation of Fasciola spp. from wild boar in Iran.