Background: The public health importance of human fascioliasis has increased during last few decades due to the appearance of new emerging and re-emerging foci in many countries. Iran, as the most important focus of human disease in Asia, has been included among six countries known to have a serious problem with fascio­liasis by WHO. Various aspects of the disease in Iran are discussed in this review.

Methods: This narrative review covers all information about human and animal fascioliasis in Iran, which has been published in local and international journals from 1960 to 2014 using various databases including PubMed, SID, Google Scholar, Scopus, Science Direct.

Results: During the period of the study the infection rates of 0.1% to 91.4% was noted in various livestock. Despite the higher infection rates of livestock in south­ern areas in past decades, human disease has been mostly encountered in northern Provinces especially in Guilan. Recent studies indicate noticeable decrease in preva­lence rates of veterinary fascioliasis in Iran, however the prevalence rates of fascio­liasis in livestock in northern Provinces of Guilan and Mazandaran seem to remain at a higher level in comparison to other parts. New foci of the disease have also been reported recently. 

Conclusion: While the prevalence of animal fascioliasis has decreased during last decades, human fascioliasis emerged as a public health problem in the country. The validity of new foci of human fascioliasis needs complementary standard studies.

Background: Echinococcus multilocularis is a tiny tapeworm, responsible for 0.3~0.5 million alveolar echinococcosis in humans.

Methods: We searched relevant papers published between 1981 and 2013 based on the database sources such as PubMed and Google scholar, and collected and integrated the data for analysis.

Results: The parasite is able to use host-originated molecules to modulate its develop­ment and has complex signalling pathways than expected previously. E. multilocularis utilizes many types of alternative splicing approaches to generate transcript isoforms. Recently, the genome of E. multilocularis has been deciphered.

Conclusion: These data will give us a profound understanding of biology of E. multilocularis, which will promote the use as a model to study helminths. 

Background: The bibliometric methods have been used in many disciplines of sciences to study the scientific production and research trends. In this study, they were used to investigate research trends related to the risk assessment of Cryptosporid­ium pathogen in water field.

Methods: Data were obtained on the Scopus database from 1993 to 2013. Re­search tendency was investigated by analyzing the distribution of languages, coun­tries, journals, author keywords, authorship pattern and co-authorship relations.

Results: The English language was dominant language of all publications (96.36%). Number of articles in this field increased from 2 in 1993 to 29 papers in 2007 and then received to 19 at the end of 2013. United States produced 35.41% of all perti­nent articles followed by United Kingdom with 10.76% and Australia with 9.92%. Water Research Journal published the most papers in this field, taking 11.62% of all, followed by Journal of Water and Health (10.92%) and Water Science and Technology (10.21%). The most productive authors were Ashbolt NJ form Canada that accounts about 1.51% of the total publications followed by Rose JB and Haas CN from United States. Authorship pattern analysis results show that literature does follow Lotka’s law (P=0.627).

Conclusion: A downward trend in the number of publications is likely to occur in future. The results of this bibliometric analysis may help relevant researchers realize the scope of the microbial risk assessment research of Cryptosporidium, and establish the further research direction.

Background: Canine visceral leishmaniasis (CVL) is not only an emerging veterinary concern but also a public health threat in endemic areas. The aim of this study was to assess the efficacy, immunogenicity and safety of two doses of aluminum hydroxide (alum) precipitated Leishmania major (Alum-ALM) mixed with BCG plus imiquimod against CVL.

Methods: A total of 560 ownership dogs were serologically tested and 234 healthy dogs with no clinical signs of CVL, no anti-Leishmania antibodies and negative leish­manin skin test were selected and double-blind randomly injected intradermally either with 0.1 ml Alum-ALM (200µg protein) mixed with BCG (2 × 10⁶ CFUs) plus imiquimod (121 dogs) or with 0.1 ml of normal saline (113 dogs).

Results: The follow-up examinations showed that there was no side effect associated with the vaccination except one case. Strong skin test conversion were seen in vac­cinated group (30.3%) compared to the control group (6.6%) at 22-24 weeks after the booster injection (p<0.001). The seroconversion was 16.3% (18/110) in vaccinated group and 26.4% (28/106) in control group after two transmission cycles but the differ­ence was not significant (P=0.095). The efficacy rate based on seroconversion was 40.4 %.

Conclusion: Two injections of Alum-ALM mixed with BCG and imiquimod is safe, although decreases the seroconversion rate of CVL, but the overall efficacy was low.

Background: Visceral leishmaniasis is systematic serous parasitic disease with pub­lic health importance. Zoonotic form of visceral leishmaniasis is wide spread in Mediterranean basin and South America regions. Direct agglutination test (DAT) is an accurate, reliable and non-expensive serological test for the diagnosis of visceral leishmaniasis in human and canines but the antigen preparation involves some limita­tions. This study aimed to compare the conventional production of DAT anti­gen with our modified DAT antigen and then assessed on human and dog pooled sera.

Methods: Conventional DAT antigen has been prepared at the School of Public Health, Tehran University of Medical Sciences and some modifications were car­ried out on it, which named as modified DAT antigen. Three positive and one nega­tive human and dog pooled serum were separately used for the comparison of modified DAT with conventional DAT antigen batches with one-month interval for a period of 9 months.

Results: A good concordance was observed between modified DAT compared to conventional DAT antigens for the detection of visceral leishmaniasis on human (100%) and dog (94.4%) pooled sera, respectively.

Conclusion: Since the modified DAT antigen could be reduced the preparation time from 3 days to several hours and a good degree of agreement was found between modified DAT and convention DAT antigen batches, it can be used as a simple and easy tool for screening and serodiagnosis of human and canine L. infan­tum infection.

Background: Viscerotropic leishmaniasis caused by Leishmania tropica poses a significant prob­lem in the diagnosis and treatment management. Since differential gene expression is more im­portant in outcome of the infection, we employed proteomic approach to identify potential pro­teins involved in visceralization of L. tropica.

Methods: The proteomes profiling of L. tropica isolated from cutaneous and visceral tissues of one host were compared by 2-DE/MS proteomics study. Moreover, the transcript level of some identified proteins was confirmed using real-time RT-PCR.

Results: Of the 700 protein spots that were detected reproducibly on each gel, 135 were found to be differentially expressed (P≤ 0.05). Most of responsive proteins in visceral isolate changed in less abundant compared to cutaneous isolate. Among differentially expressed proteins, 56 proteins were confidently identified and classified according to the biological process. The larg­est groups consist of proteins involved in carbohydrate metabolism and protein synthesis. Most of the identified proteins, which implicated in energy metabolism, cell signaling and virulence were down-regulated, whereas some proteins that have a role in protein folding, antioxidant defense and proteolysis were up-regulated in visceral form. Moreover, the transcript level of some identified proteins such as co-chaperon was confirmed using real-time RT-PCR.

Conclusion: L. tropica probably uses different mechanisms for survival and multiplication in viscera to establish viscerotropic leishmaniasis. The current study provides some clues into the mechanisms underlying the dissemination of L. tropica.

Background: Microsporidia species are obligatory intracellular agents that can in­fect all major animal groups including mammals, birds, fishes and insects. Whereas world­wide human infection reports are increasing, the cognition of sources of infec­tion particularly zoonotic transmission could be helpful. We aimed to detect zoono­tic microsporidia spore in fecal samples from some animals with close – contact to human.

Methods: Overall, 142 fecal samples were collected from animals with closed-con­tact to human, during 2012-2013. Trichrome – blue staining were performed and DNA was then extracted from samples, identified positive, microscopically. Nested PCR was also carried out with primers targeting SSU rRNA gene and PCR products were sequenced.

Results: From 142 stool samples, microsporidia spores have been observed microscopi­cally in 15 (10.56%) samples. En. cuniculi was found in the faces of 3 (15%) small white mice and 1 (10%) laboratory rabbits(totally 2.81%). Moreover, E. bieneusi was detected in 3 (10%) samples of sheep, 2 (5.12%) cattle, 1 (10%) rabbit, 3 (11.53%) cats and 2 (11.76%) ownership dogs (totally 7.74%). Phylogenetic analysis showed interesting data. This is the first study in Iran, which identified E. bieneusi and En. Cuniculi in fecal samples of laboratory animals with close – contact to human as well as domesticated animal and analyzed them in phylogenetic tree.

Conclusion: E. bieneusi is the most prevalent microsporidia species in animals. Our results can also alert us about potentially zoonotic transmission of microsporidiosis.

Background: Interleukin 18 (IL-18) exerts pleiotropic roles in many inflammatory-related diseases including parasitic infection. Previous studies have demonstrated the promising therapeutic potential of modulating IL-18 bioactivity in various pathologi­cal conditions. However, its involvement during malaria infection has yet to be established. In this study, we demonstrated the effect of modulating IL-18 on the histopathological conditions of malaria infected mice.

Methods: Plasmodium berghei ANKA infection in male ICR mice was used as a model for malaria infection. Modulation of IL-18 release was carried out by treat­ment of malarial mice with recombinant mouse IL-18 (rmIL-18) and recombinant mouse IL-18 Fc chimera (rmIL-18Fc) intravenously. Histopathological study and analysis were performed on major organs including brain, liver, spleen, lungs and kidney.

Results: Treatment with rmIL-18Fc resulted in significant improvements on the histopathological conditions of the organs in malaria-infected mice.

Conclusion: IL-18 is an important mediator of malaria pathogenesis and targeting IL-18 could prove beneficial in malaria-infected host.

Background: This study was conducted to investigate the influence of Toxoplasma gondii infection on spermatic and hormonal parameters in a pilot sample of immunocompe­tent human male subjects.

Methods: This cross sectional, observational pilot study on 60 immunocompetent hu­man male subjects aged between 18 and 60 yr old was conducted between 2012 -2013. Blind evaluation of serological markers of past T. gondii infection (TOX-IgG, TOX-IgM) was performed, along with individual spermiograms and determinations of folli­cle-stimulating hormone (FSH) and testosterone serum levels.

Results: The overall prevalence of past T. gondii infection in the investigated immunocom­petent male subjects was 25%. No statistically significant influence of T. gondii infection on sperm characteristics (ejaculate quantity, sperm count, motility, morphol­ogy) and serum levels of FSH or testosterone were found. Among possible predictors of a modified spermiogram studied by multiple logistic regression along with the T. gondii infection (age, smoking, alcohol consumption, fertility influencing malfor­mations, infections, trauma or medication), only the presence of varicocele in the medi­cal history of the studied subjects was found to significantly participate in the predic­tion of a modified spermiogram (P=0.0154). A necessary sample size of 994 subjects was computed in order to achieve a test power of 0.8 (80%) to discriminate an effect size of 8.89% estimated by our pilot study.

Conclusions: Although our investigation did not demonstrate an influence of latent T. gondii infection on spermatic and hormonal parameters of immunocompetent male hu­mans, the absence of such an influence cannot be affirmed, due to the limited sample size of our pilot study. 

Background: In this study morphological and molecular characterization of Acan­thamoeba strains, isolated from dental unit waterlines (DUWLs) were surveyed and the levels of disinfection achievable in vitro by the application of ozone disinfect­ant to DUWLs were evaluate.

Methods: Water samples were collected from air-water syringes, cup fillers and tap water before and at the end of the working day. They were cultured on non-nutrient agar (NNA) plates. Species identification was carried out with a PCR assay based on sequence analysis of the 18S rRNA gene. The cellular response to ozone was tested on Acanthamoeba cyst with different doses at different contact time in vitro twice.

Results: Prevalence rates for Acanthamoeba contamination were 100, 100 and 72% for air-water syringes, cup fillers and tap water, respectively. The morphological analysis revealed the presence of A. castellanii, A. griffin, A. hatchitti and A. lenticulata. Phylogenetic analysis of the sequences showed the four strains to be closely related to a sequence type (T3, T4, T5 and T11). Acanthamoeba cells were stained with try­pan blue, which revealed killed of Acanthamoeba instantaneously after 10 minutes in ozonized water. There was no growth of Acanthamoeba occurred after ozone treat­ment in water bottles for 5 minutes with a flow rate of 500 mg/hour.

Conclusion: Ozone can play an important role in controlling the problem of contami­nation of DUWLs as a potent disinfectant.

Background: The aim of this study was to determine phenolic acid composi­tion and anti-parasitic effects of Peucedanum caucasicum, P. palimbioides, P. longibracteola­tum and P. chryseum on Entamoeba histolytica.

Methods: Methanol extracts of the plant species were prepared by soxhlet extrac­tion. Phenolic acid compositions were determined by HPLC. Anti-prolifera­tive effect of extracts on trophozoites was determined by using trypan blue dye exclusion test. For counting the cells, approximately a hundred of E. histolytica trophozoites were examined in each time. The data were presented as mean values with standard deviations and analyzed by repeated measures of ANOVA followed by Tukey test for post-hoc pairwise comparisons. The P-value was set at 0.05 for significance level.

Results: All of the extracts showed a time and dose dependent amoebicidal ac­tion on trophozoites. Among the extracts tested, P. longibracteolatum showed the strongest amoebicidal effect on the trophozoites. As expected, this plant species also exhibited time and dose dependent activity on the trophozoites. At 4.0 mg/ml extract concentration, all of the trophozoites were killed by the extract in 72^nd hour. Gallic (11.144 mg/g), P-hydroxybenzoic (17.646 mg/g), and o-couma­ric acids (14.442 mg/g) were determined as the major phenolic acids of P. longibracteo­latum. Gallic and P-hydroxybenzoic acids found in P. longibracteolatum could not be determined in other extracts. Therefore, high activity potential of this plant could probably be attributed to the presence of these phytochemicals.

Conclusion: P. longibracteolatum can be further evaluated as potential therapeu­tic drugs for the treatment of Entamoeba infections.

Background: The present study aimed to determine the prevalence and associated risk factors of vaginal trichomoniasis in women referred to gynecologic clinic in Benha University Hospital, Egypt.

Methods: Two hundred female patients enrolled in the study. Vaginal samples were obtained from them and examined for T. vaginalis by wet mount, Giemsa stain, Acridine orange (AO) stain and culture on modified Diamond’s medium. For analysis of accuracy of the methods used, the receiver operating characteristic (ROC) curve concept with culture as a gold standard was applied.

Results: Out of 200 patients, T. vaginalis was found in 22 (11%) patients by any of the diagnostic methods used. The accuracy of AO staining comes next to Dia­mond’s culture (AUC 0.909, sensitivity 81.8%, specificity 100%, CI 0.81-1.0) fol­lowed by Giemsa staining (AUC 0.835, sensitivity 68.2%, specificity 98.9%, CI 0.72-0.95). The wet mount was the least accurate method (AUC 0.795, sensitivity 59.1%, specificity 100%, CI 0.67-0.92). There was no significant association be­tween potentially supposed risk factors and trichomoniasis except patients complain­ing of either dysuria and dyspareunia or back pain and abdominal pain.

Conclusion: Trichomoniasis is a common disease in our community. Sociodemo­graphic factors do not seem to affect the prevalence among different Egyptian population. For accurate diagnosis, laboratory investigation is essential. A positive wet smear is diagnostic, but negative samples should be examined by methods that are more sensitive.

Background: Cryptosporidium is an intestinal protozean parasite causing water­borne and foodborne outbreaks of diarrheal diseases. The present study was per­formed in order to find prevalence and subtypes of Cryptosporidium among children with diarrhea in Gonbad Kavoos City, Northern Iran.

Methods: Diarrheic samples were collected from 547 children. The initial parasitologi­cal diagnosis was made based on detection of oocysts using the modi­fied Ziehl-Neelsen acid-fast staining method. The positive microscopically samples were selected for sequence analysis of partial 60 kDa glycoprotein (gp60) gene.

Results: Out of 547 collected samples, 27 (4.94%) were positive for Cryptosporid­ium oocysts. Fifteen from 27 positive samples successfully amplified in PCR. Se­quences analysis of gp60 gene in 15 Cryptosporidium isolates revealed that all of them (100%) were C. parvum. The results showed three subtypes of IIa subtype family (7 cases) including IIaA16G2R1, IIaA17G1R1, IIaA22G3R1 and one subtype of IId subtype family (8 cases). The most common allele was IId A17G1d (53.3%).

Background: As for human trichomoniasis the host-parasite relationship is very complex, and the broad ranges of clinical symptoms are unlikely be attributable to a single pathogenic mechanism. Specific Random Amplified Polymorphic DNA (RAPD) markers of 490 bp, 720 bp and 460 bp using the primers Tv-5, OPA-6 and OPA-11, respectively, were reported. This was the first description of possible ge­netic virulence markers of the infection by T. vaginalis. The aim of this study was to characterize the specific RAPD markers in order to elucidate their importance on virulence of this illness.

Methods: The selected specific RAPD fragments were cloned and sequenced. The obtained sequences were compared by the BLAST algorithm.

Results: The nucleotide sequence of the Tv-5₄₉₀ RAPD marker exhibited signifi­cant similarity to T. vaginalis hypothetical G3 leucine rich repeat (LRR) family pro­tein (e-value: 6e-14) and Giardia lamblia leucine rich repeat protein 1 virus receptor protein (e-value: 6e-14 and 2e-12) ; however, the OPA-6₇₂₀ and OPA-11₄₆₀ showed no significant similarity with any coding published sequence. All the evaluated strains showed the presence of the LRR gene.

Background:Natural killer (NK) cells play an important role in early stages of innate immune responses against viral and tumoral attacks. Activation of NK cells by leishmaniasis results in secretion of cytokines such as interferon (IFN)-γ and tumor necrosis factor (TNF)-α, which enhance the phagocytosis and clear­ance of parasite. Lipophosphoglycan 3 (LPG3), the Leishmania homologous with GRP94 (glucose regulated protein 94), a member of HSP90 family, contributes to LPG assembly as the most abundant macromolecule on the surface of Leishmania promastigotes.

Methods:We purified NK cells from healthy individuals (n=10) using magnetic-activated cell sorting (MACS) technology. Purified NK cells were co-incubated with different concentrations of recombinant LPG3 (rLPG3), and its N-terminal (NT) and C-terminal (CT) fragments. Finally, the production of IFN-γ and TNF-α by NK cells were measured by ELISA.

Results:Recombinant LPG3 but not its fragments (CT and NT), could signifi­cantly enhance the production of TNF-α by NK cells (P<0.05). Moreover, rLPG3, CT, and NT fragments were markedly stimulated the secretion of IFN-γ by NK cells (P<0.001).

Background: Epidemiological investigations on Toxoplasma gondii infection have found a significant association between human toxoplasmosis and consumption of raw or undercooked meat. The present study aimed to characterize genotypes of T. gondii in 20 cattle, 40 sheep, 15 goats and 16 pigs from the North of Portugal.

Methods: Nested PCR amplified the surface antigen 2 (SAG2) gene. Sequencing analy­sis was performed in order to assess the prevalence of SAG2 type strains (I, II and III).

Results: Three and 4 strains of SAG2 type II were identified in heart samples of cattle and sheep, respectively. Three SAG2 type II strains were detected in brain, diaphragm and heart of 3 pigs. Three strains detected in heart samples of 3 goats belonged to SAG2 types I or II; with the same result being observed in heart samples from 2 sheep and in 2 brain and 1 heart samples from 3 pigs.

Background:Human toxocariasis is a parasitic infection caused by the larvae of Toxocaracanis. We examine the Toxocara seroprevalance in veterinarians and animal husbandry workers living in the Mugla Province, Turkey to evaluate better the risk factors for Toxocara exposure.

Methods: In 2014, 376 volunteers participated in the study in 2014. All blood specimens were tested using a commercial enzyme immunoassay kit and ELISA positive samples were confirmed by Western Blot (WB) method.

Results: The seroprevalence of Toxocara, as determined by WB, was 8%. A statistically significant correlation was evident between patient age and Toxo­cara positivity among animal husbandry workers (P = 0.029). A strong associa­tion was also evident between sex and seropositivity in the animal husbandry group (P=0.024). Veterinarians working in pet clinics did in fact exhibit higher Toxocara seropositivities relative to those of other groups (P = 0.029). A statisti­cally significant difference was detected between the rural geographic areas surveyed (P = 0.04).

Conclusion: In Mugla Province, seroprevalence of Toxocara is lower than other regions. Despite the low seroprevalence observed, especially in high risk professions toxocariasis remains an important medical concern within the region. 

Background: Infection by Toxocara spp. is known to be significantly associated with partial epilepsy. It has become popular for people to raise dogs/cats as pets and consume roasted meat/viscera, and the status of Toxocara spp. infection, epi­lepsy awareness, and associated risk factors among the general population are cur­rently unknown in Taiwan.

Methods: A seroepidemiological investigation among 203 college students (CSs), consisting of 110 males and 93 females with an average age of 21.5 ± 1.2 years, was conducted in 2009 in Taipei City. A Western blot analysis based on excretory-secre­tory antigens derived from Toxocara canis larvae (TcESs) was applied to determine the positivity of serum immunoglobulin G antibodies. A self-administered question­naire was also given to obtain information about demographic characteris­tics, epilepsy awareness, and risk factors. A logistic regression model was applied for the statistical analysis using SPSS software.

Results: The overall seropositive rate of Toxocara spp. infection was 8.4% (17/203). As to epilepsy awareness, a non-significantly higher seroprevalence was found in CSs who claimed to "know" about epilepsy compared to those who did not know (P > 0.05).

Conclusions: It appears that appropriate educational programs are urgently needed to provide correct knowledge related to the prevention and control measures against Toxocara spp. infections to avoid potential threats by this parasite to the general population in Taiwan.

Background: Trichomonas vaginalis, the causative agent of trichomoniasis, is responsible for more than half of all sexually transmitted infections (STIs). The present study aimed to deter­mine the frequency of T. vaginalis infection and its clinical manifestations in sympto­matic pregnant women in the area based on four different diagnostic methods.

Methods: A total of 162 pregnant women with at least one sign or symptom of vaginosis, referred to two gynecologic and obstetrics clinics in Rafsanjan City, south central Iran, were randomly selected in 2012-13. Through speculum examination of patients by gynecologists, clinical diagnosis determined, vaginal discharge were collected by using two sterile cotton swabs from the posterior fornix and vagina pH was measured. Samples were examined by three diagnostic methods including wet mount, culture in TYI-S-33 medium and polymerase chain reaction (PCR).

Results: T. vaginalis was detected in 19.5%, 27.2%, 56.2% and 51.6% of subjects according to diagnostic methods of clinical diagnosis, wet mount, culture and PCR, respectively. There was statistically significant relationship between T. vaginalis infection and patients' age, gestational age, marriage age, residence, educational level, parity. The symptomatological pattern in the 91 women infected with T. vaginalis was as follows: leukorrhea, 96.7%; urine frequency, 65.9%; odorous secretion, 63.3%; urogenital itching and irritation, 53.8%; vagi­nal inflammation, 47.3%; dyspareunia, 39.6%; and dysuria, 16.5%.

Background: Recognition of helminth-derived pathogen associated molecular patterns (PAMPs) by pattern recognition receptors (PRRs), including toll like recep­tors (TLRs) is the first step towards initiating anti–helminth immune re­sponses.

Methods: Using somatic antigens of Ostertagia occidentalis, an important abomasal parasite of ruminants, the expression of ovine TLR2 and TLR4 in peripheral blood mononuclear cells (PBMCs) was analyzed by real-time quatitative reverse-transcrip­tion polymerase chain reaction (qRT-PCR). Somatic antigens of O. occidentalis were prepared to stimulate ovine PBMCs in a time and dose dependent manner.

Results: A high expression of TLR2 and TLR4 was observed in PBMCs cultured with somatic antigens of the parasites specially when PBMCs were cultured with 100 µg/ml of somatic antigens and incubated for 2h. Up-regulation of TLR2 expres­sion was more pronounced and evident in our study.