Background: Toxoplasma gondii, the coccidian protozoan parasite with worldwide distribution, is the agent of toxoplasmosis. The disease is life threatening in congenital form and in immunocompromised patients. The present study was carried out in 2016 to evaluate the in vitro effects of nanosilver colloid on tachyzoites and bradyzoites of T. gondii, RH and Tehran strains.

Methods: Different concentrations (5, 10 , 20 ppm) of  nanosilver colloid were added to tachyzoites of T. gondii , RH strain (type I) and bradyzoites and tissue cysts of T. gondii , Tehran strain (type II) and incubated for 30, 60, 90 and 120 minutes. The mortality rates of tachyzoites and bradyzoites were evaluated by trypan blue dye and MTT assay. Then SEM carried out to show the changes between control and exposed parasites.

Results: The greatest mortality rate was seen in 20 ppm concentration and after 120 minutes of exposure. By electron microscopy, the structural changes were seen in tachyzoites of RH and tissue cyst of Tehran strain in comparison with control groups.

Conclusion: Nanosilver colloid was effective on both tachyzoites and bradyzoites of T. gondii, RH and Tehran strains.

Background: The protozoan Giardia and Cryptosporidium are responsible for most water-borne diseases all over the world. The extent and number of outbreaks of waterborne diseases suggests a significant risk of their potential transmission via drinking water. This study aimed to document the prevalence and viability of Giardia and Cryptosporidium (oo) cysts in tap water samples in Beni-Suef Governorate, Egypt and to detect the predominant Giardia and Cryptosporidium assemblages/species using nested PCR/ Restriction Fragment Length Polymorphism (RFLP) confirmed by further sequencing of positive samples.

Methods: A total of 80 tap water samples were collected throughout a year from four big centers and filtered using the membrane filtration method. Samples were stained by Lugol’s iodine, Modified Zeihl-Neelsen (MZN) (to detect prevalence) and trypan blue stain (to detect viability). Nested PCR-RFLP and sequencing were used for molecular characterizations and genotyping of the detected Giardia and Cryptosporidium.

Results: Giardia and Cryptosporidium DNA was detected in 20 (25%) and 29 (36.3%) samples respectively, with predominance of Giardia assemblage B (85%) and C. hominis (75.9%). The prevalence and viability of both parasites (oo) cysts showed seasonality which peaked in summer and were greater in Beba center and in rural areas.

Conclusion: To our knowledge, no studies have been done in these areas before. The anthroponotic transmission has an important role in giardiasis and crypto­sporidiosis epidemiology in this studied area.

Background: The aim of this research was to compare excretory-secretory and somatic extract materials of Fasciola hepatica and F. gigantica to detect protein maps of two species.

Methods: Twenty infected livers were collected from sheep in industrial slaughterhouse in Tehran, 2017-2019. Worms were detached from bile ducts, then recognized according to morphologic and morphometric criteria. After three times washing, worms were incubated in RPMI culture media and excretory-secretory products were collected. Worms were crushed and homogenized for preparation of somatic extract. Two Dimensional Electrophoresis gels were accomplished for both excretory-secretory material and somatic extracts. Gels were scanned with densitometer and analyzed with Same Spots software and protein spots were identified with Expasy database.

Results: For both excretory-secretory products and somatic extract, protein spots were appeared with two-dimensional electrophoresis technique. Quantitative analysis showed 40 and 28 protein spots for excretory-secretory of F. hepatica and F. gigantica respectively. For somatic extract 19 and 12 protein spots were recognized for F. hepatica and F. gigantica in that order.

Conclusion: The rate of expression of some proteins were more in F. hepatica while expression of other proteins was high in F. gigantica. The expression of protease enzyme was higher in F. gigantica than F. hepatica. These data could be considered for biochemical differentiation of Fasciola species and subsequently to design and prepare of antigens for diagnosis/vaccine development.

Background: Fascioliasis is one of the most important food-borne worm disease caused by Fasciola sp. Parasitological diagnosis is more difficult due to the low parasite burden and a few eggs shedding of helminths. Therefore, it will be valuable to development of simple, fast and reliable diagnostic tests for detection of human and animal fascioliasis.

Methods: Infected liver collected from abattoir in Tehran, Iran in 2017. F. hepatica eggs were detached from the uterus of worms under a stereo microscope. Various numbers of eggs were spiked to 200 mgr. of negative feces samples. DNA was extracted and then target regions (nuclear IGS) were amplified by LAMP assay using six primers. Fecal specimens without egg and DNA of other helminths were used as negative controls. F. hepatica sample which confirmed by morphologic criteria and PCR- RFLP was used as positive control.

Results: LAMP products by using SYBR Green I could detect even a single egg in fecal samples which was visible by change of color from orange to green. There was no cross amplification by other helminths including; Taenia saginata, Dicrosolium dendriticum and F. gigantica.

Conclusion: LAMP seems a rapid, sensitive, cost-effective technique for detection of human fascioliasis.

Background: In this research, the effect of morphine on promastigotes and amastigotes of Leishmania major has been investigated in the presence of nalmefene as a blocking opioid drug and imiquimod as an opioid growth factor receptor.

Methods: This study was conducted at Tarbiat Modares University, Tehran, Iran in 2015-2018. Morphine with different concentration (0.1, 1, 10 and 100 1µg/ml) alone and with imiquimod (0.01, 0.1 and 1µg/ml) and nalmefene (0.1, 1 and 10 µg/ml) on promastigotes and amastigotes in macrophages and also the percentage of infected macrophages was investigated. For evaluation of the apoptosis, we used flow cytometry method. The effect of imiquimod and nalmefene on glucantime and amphotericin B as current drugs for treatment of leishmaniasis was evaluated too.

Results: The effect of morphine on promastigotes and amastigotes has a reverse relationship with its concentration. The results of flow cytometry for drug-treated promastigotes revealed that apoptosis and necrosis did not increase markedly relative to the control group. A combination of morphine and imiquimod in concentrations of 0.05, 5 and 5 µg/ml had a pronounced effect on reduction and prevention of macrophage infection with amastigotes. Morphine at a concentration of 0.1 µg/ml plays the role of adjunctive treatment. In amastigote assay we found the better results in group that get glucantime 25 µg/ml+ imiquimod 0.5 µg/ml.

Conclusion: This effect is strengthened with imiquimod and weakened with nalmefene. Using high dose morphine and nalmefene had reverse effects. They suppress immune system and had no controlling effect in macrophages amastigote infection and reduction of promastigotes.

Background: Equine piroplasmosis is a major tick-borne disease that can lead to serious health problems and economic losses in horse industry. The aim of the study is to determine the prevalence of Theileria equi and Babesia caballi in different species of Equus namely (Horse, mule, donkey and pony) by Giemsa stained blood films and competitive ELISA.

Methods: This study was conducted at various geographic areas of Erbil governorate.  A total of 349 blood samples from equine (209 horses, 62 mule, 57 donkey and 21 ponies) were collected to estimate the prevalence rate of Theileria equi and Babesia caballi by using Giemsa stained blood smear and competitive ELISA.

Results: The overall prevalence rates were (10.6%) consisting of (8.3%), (1.7%) and (0.6) for T. equi, B. caballi and both infection respectively by Giemsa stained blood smears, and the rate was 38.97%, consisting of 20.9% for T. equi, 11.2% for B. caballi and 6.9% for both infection by cELISA. Seroprevalence rate of T. equi was significantly higher (P < 0.001) than that B. caballi in equids. There was also a significant difference associated between age (P < 0.01), activity (P < 0.01), management (P < 0.001) and tick infestation (P < 0.001), but neither the type of equids nor the gender was significant differences associated with prevalence rate.

Conclusion: The present study concluded that the equine piroplasmosis is a vital infection distributed among the equine in Erbil province North of Iraq. Thus a better control programme should be implemented to contain and control the prevalence of the disease within the area.

Background: The aim of the present study was to study pathological and parasitological characteristics of simultaneous capillariasis and hystrichiasis in a diseased Anas crecca captured in Mazandaran Province of Iran on Oct 2016.

Methods: Parasitology and histopathology techniques were used to detect the parasites of the carcass of the captive common teal.

Results: Macroscopically, severe chronic inflammatory reactions and nodular or granuloma formation and irregular thickening of the affected wall of the esophagus, crop, proventrculus and ventriculus were observed. Microscopically, apart from the chronic inflammatory reactions and granuloma formation, in the paraffin sections stained with Harris Hematoxyline and Eosin technique, the characteristics of the mature female and male Capillaria spp.  and their barrel-shaped operculated embryonated eggs were found within the tunnels burrowed by the nematode in the hyperplastic stratified squamous epithelium of the inflamed crop and distal portion of the esophagus. The mature female Hystrichis spp. containing oval-shaped, embryonated non-operculated eggs and male parasite were found within the labyrinthus spaces formed in the submucosa granulomas of the ventriculus and proventriculus.

Conclusion: Based on the parasitological and pathological studies, the species of the nematode parasites were identified as Capillaria (Thominx) contorta and Hystrichis. tricolor.

Background: Most of trematodes need snails to complete their life cycles. Consequently freshwater snails are served as intermediate hosts of many parasites worldwide. There is a lack of report on snail diversity and parasitic infection in snails in Sirindhorn Reservoir, Ubon Ratchathani province, Thailand.

Methods: Mollusk diversity and trematode cercariae infections were investigated in snails from 120 sampling sites surround Sirindhorn Reservoir from April 2018 to June 2018. Mollusk species were identified based on their shell morphology. The presence of cercariae infections in snails was examined by cercarial shedding methods. The interaction between snail species was analyzed by the correlation method.

Results: Overall, 2076 mollusks were collected which comprised six species of snails and two species of bivalves. Snail species were identified as Bithynia siamensis goniomphalos, Anentome helena, Filopaludina sumatrensis spiciosa, F. martensi martensi, F. martensi munensis and Pomacea canaliculata. The overall rate of trematode cercariae infection was 1.69% (35/2,076). The cercariae found infecting snails were Cercariaeum cercaria, Virgulate cercaria, Cotylomicrocercous cercaria and Furcocercous cercaria. The most common snails found was the assassin snail, A. helena, which showed the negative relationship among other species interactions.

Conclusion: This finding indicated infection with animal’s parasites in snails in this area are common, besides, we found many species of snails in Sirindhorn Reservoir are potentially be the host of parasite in animal and human.

Background: One of the severe complications of toxoplasmosis is the induction of abortion in sheep, goats, and human beings. The rate of abortions related to toxoplasmosis in sheep varies between 10% and 23% in the United States and European countries. In Iran, Toxoplasma infection were diagnosed in aborted ovine fetuses between 13.5% and 69% in the brain samples based on PCR. The purpose of this study was to investigate the prevalence of abortions related to T. gondii among the aborted fetuses of sheep in Ardabil area, North-West of Iran.

Methods: The brains of 75 aborted sheep fetuses in Ardebil area were investigated with nested PCR method using fragments of the GRA6 gene during lambing seasons in 2014-2015. Meanwhile, thoracic and abdominal fluids of aborted fetuses and 200 serum samples collected from the herds affected by abortion were examined for the existence of anti-T. gondii IgG.

Results: For 48 out of the 75 fetal brain samples (64%), the results of nested PCR test were found to be positive. Furthermore, antibodies against Toxoplasma was observed in 136 (68%) collected samples of sheep and 21 samples (28%) of fetal fluids.

Conclusion: There is the significant role of T. gondii in the abortions of sheep in Ardabil area. Meanwhile, this condition can also be dangerous for human beings because of their consumption of sheep meat.

Background: Hydatidosis is a neglected global zoonotic disease, caused by larval stage of the cestode Echinococcus granulosus in human and animal. Because of high economic and medical importance of the disease, this study was performed to find the seroprevalence of human hydatidosis in Gorgan City, Golestan Province, northern Iran.

Methods: In this cross-sectional study, blood samples were collected from people referring to Reference laboratory of Golestan University of Medical sciences in 2017. A relevant questionnaire was completed for demographic data for each person. Echinococcus IgG antibody was investigated by ELISA using native antigen B. The data were analyzed using SPSS software applying logistic regression.

Results: Overall, 612 blood samples were collected. Cut-off value was considered 0.29. Sixteen cases (2.6%) were seropositive for hydatidosis. The seroprevalence of hydatidosis was 2.3% and 4.7% among males and females, respectively. There was no statistically significant correlation between the hydatidosis and investigated variables such as sex, age, tribes, residence, education, etc.

Conclusion: The prevalence of human hydatidosis shows approximately the same range as other regions of Iran. Although due to the neighboring the Mazandaran Province reported as the highest seroprevalence of hydatidosis, we expected more rate of seropositivity.

Background: Malaria eradication has been complicated by the repeated emergence of antimalarial drug resistances. We aimed to determine whether a metabolite extract of Streptomyces hygrocopicus subsp. hygroscopicus could decrease the viability of Plasmodium falciparum 3D7 in vitro.

Methods: S. hygroscopicus subsp. hygroscopicus isolates were inoculated and fermented on the ISP4 medium. The fermented S. hygroscopicus was mixed with ethylacetate 1:5 (v/v), and the solvent phase was evaporated. Several concentrations of isolated extract was added to the P. falciparum 3D7 culture containing trophozoite and schizont stages in 24 wells plates when the degree of parasite-infected erythrocytes reached 5%, then incubated for 8 hours. DNA parasite density was measured using flow cytometry, parasite degree and morphology were observed under microscopic by Giemsa-stained smears. 

Results: The metabolite extract affected the morphology of almost all of parasite asexual stages. Schizonts and trophozoites failed to grow and appeared damaged with pycnotic cores and loss of cytoplasmic content. At 8 hours there was a significant decrease in DNA parasite density in culture exposed to 2.6 mg/ml and 13 mg/ml (P = 0.002; P = 0.024) of the extract. The degree of parasite-infected erythrocytes was decreased from the beginning of exposure (0.02 mg/ml of the extract). There was a significant inverse correlation between the concentration of extract and the degree of parasite-infected erythrocytes as well as the density of DNA parasite (r = -0.772, P = 0.000; r =-0.753; P =0.000).

Conclusion: Metabolite extract of S. hygroscopicus subsp. hygroscopicus causes morphological damage, decreases the degree of parasite-infected erythrocytes and the DNA density of P. falciparum 3D7 in vitro.

Background: Eye diseases, including, Uveitis, inflammation of the retina, and choroid are caused by different agents in humans and has a variety of anterior medial and posterior types. The agent of eye diseases is very different from simple bacterial to acute viral, fungal and parasitic infection. There is limited information regarding the type of eye diseases and Toxocara canis.

Methods: Blood samples of 359 individuals (339 patients including endophthalmitis, uveitis, DCR, glaucoma, and cataract 20 individuals as control group) together with their information were collected from ophthalmology hospitals in Tehran, Iran from Feb 2013 to Jan 2015. The patient's serum was evaluated for the presence of anti-Toxocara antibodies by ELISA kits and blood smears for high eosinophilia. The positive samples were confirmed by Western blot analysis for T. canis infection.

Results: Overall, 339 patients sera with eye diseases and control group were tested for anti-T. canis antibodies. Nineteen (5.6%) patients had anti-T. canis IgG that 14 (6.1%) were male and 5 (4.5%) were female, and all the patients had negative eosinophilia as well. The results of Western blot analysis for 19 positive patients indicated that 15 were infected by T. canis and 4 were infected by other parasitic infection. The results for control group were negative.

Conclusion: All the patients with inflammatory eye diseases such as endophthalmitis, uveitis DCR, glaucoma and cataract were studied for Toxocara infection in this research work were at risk. Therefore, in contrast to the previous idea, all eye inflammatory diseases in ocular patients should be considered for Toxocara antibodies in addition to Uveitis.

Background: Demodex mites are permanent ectoparasites of human pilosebaceous unit. They mainly infect skin of the face and scalp. Many studies have shown higher density of the ectoparasites in diseased inflammatory skin than in normal skin. The aim of this study was to determine the frequency of Demodex folliculorum (DF) in treatment-resistant patients with the combination of allergic rhinitis (AR) and diabetes mellitus (DM).

Method: This study was conducted in 2014-2017. It included 92 patients aged 18-70 years who presented at the Ear, Nose and Throat (ENT) and Endocrinology Polyclinics of Mustafa Kemal University Medical Faculty Hospital, Turkey. An age and gender matched control group was formed of 30 healthy individuals. To determine the presence of DF, a few eyelashes were taken from eyelids in both groups. Then samples were examined under a light microscope.

Results: DF positivity was determined in 44 (47.8%) of the 92 patients and in 1 (3.3%) of the 30 control group subjects. In the patient group, DF positivity was evaluated as present in 14 (43.7%) of the DM patients, in 12 (40%) of the AR patients and in 18 (60%) of the AR+DM patients. Statistically significant DF incidence was found in all three patient groups compared to the control group (P=0.001). The incidence in AR + DM group was not different from other patient groups.

Conclusion: DM, AR, advanced age and obesity had prepared the environment for Demodex infestations. This issue should be considered especially in treatment of patients with AR+DM.

Background: The first line treatment for cutaneous leishmaniasis is pentavalent antimony such as sodium stibogluconate (pentostam) and meglumine antimonite (glucantime). One of the most important ways to uptake the drug is by a transmembrane protein, called aquaglyceroporin encoded by Aquaglyceroprotein1 (LmAQP1). In this study, molecular characterization of LmAQP1 was reported.

Methods: Leishmania major (MRHO/IR/75/ER) promastigotes were cultured, and then DNA extraction and RNA extraction were done and followed by cDNA synthesis. Amplicons resulted from PCR and RT-PCR using specific primers were purified and sequenced. Molecular characterization was done by bioinformatically software such as BLST, ClustalW2, and RMSD.

Results: Amplicons resulted from PCR and RT-PCR showed equal size in length. BLASTn analysis showed a point nucleotide change in LmAQP1 gene that encoded 282-amino-acid long protein with a mutation at position 154 including replacement of alanine by threonine. The observed mutation in the interested gene was assessed using the above-mentioned software. The mentioned gene was submitted at GenBank, NCBI with accession number of KU514052.

Conclusion: The functional prediction of the protein encoded from LmAQP1 showed that the mentioned mutation could not affect the three-dimension structure, but it may modify the drug uptake potential of this important channel. Based on from LmAQP1 role, it seems to be an appropriate candidate for drug development. According to search through internet, this is the first report of LmAQP1 from L. major (MRHO/IR/75/ER).

Cutaneous leishmaniasis (CL) is one of the prevalent parasitic diseases in Iran principally caused by two species, Leishmania major and L. tropica. Here, we present a rare case of a congenital form of CL around the glans penis from the central part of Iran in 2017. A 24-yr-old male patient from the central part of Iran presented with biennial ulceration of the glans penis. Diagnostic methods included physical and preclinical examination, microscopic observation, leishmanin skin test (LST), and serological tests including direct agglutination test (DAT). Nested PCR and sequencing analysis were used on the positive smears for confirmation of CL and Leishmania species identification. The preclinical results were normal, and no anti-Leishmania antibodies were detected in the peripheral blood of the patient using DAT. In abdominal ultrasonography, the spleen and liver size were normal. LST was positive (≥5 mm) after 72 h, and a few amastigote forms of Leishmania sp. were demonstrated under light microscopy. L. major was confirmed using nested PCR and sequencing analysis. The patient responded to oral administration of miltefosine (2.5 mg/kg/d) for 28 days. To the best of our knowledge, genital CL due to L. major has not been previously reported from Iran.

Ascariasis is an important and prevalent geo-helminth infection, especially in underdeveloped countries such as Afghanistan, where public health problems and devastation of hygienic infrastructures usually lead to the higher incidence of the infection. Here, we report two cases of intestinal obstruction that accompanied with biliary ascariasis due to Ascaris lumbercoides in pregnant women and a 6-year-old girl from Afghanistan. Biliary ascariasis can be associated with cystic duct obstruction, gallbladder dilation, acute cholecystitis, acute cholangitis, obstructive jaundice, and biliary colic.

Spirocerca lupi is an international nematode of Canine and wild carnivores. The generic clinical symptoms are regurgitation, vomiting, and pursiness. The parasite usually procreates nodular masses in the esophagus and pectoral aorta. On July 2014, a four-month male sandy fox was found accidentally killed on the road in Bafgh City, Southeast of Yazd, Yazd Province, Iran during necropsy, referring to esophagus, parasites were collected from esophagus and transmissive up to 70% of alcohol. Parasites were recognized as S. lupi. After clarification of lactophenol and staining, the nematodes were recognized as S. lupi according to key guidelines of Taylor. Besides, the tissue samples were isolated from esophagus wall - the appendix site of nematode for histopathological investigation - so they were transmissive up to 10% of buffered formalin and stained with haematoxylin and eosin. Microscopic diagnoses in esophagus were included epithelial destruction, wall thickening, inflammatory cells infiltration, necrosis and vascular thrombosis. With the author’s knowledge, this is the first report and histopathological investigation of S. lupi in sandy fox.

Hydatid cyst caused by Echinococcus granulosus usually involves lung and liver but can appear in other organs. We report a 29-yr-old woman presented to Fatemi Hospital, Ardabil, Iran in 2017 with progressive painful swelling of the left gluteus which in imaging showed hydatid cyst. The cyst was successfully en blocked and the patient was discharged on albendazole treatment with no recurrence in the symptoms during the first week, first and second months after surgery follow-up and in the final visit at third months. In the endemic regions, the possibility of hydatid cysts should be considered in differential diagnosis of any cystic mass. 

We present a 40-year-old woman with a history of relapsing echinococcosis who had undergone a number of surgical procedures for cyst removal (right pulmectomy, cardiac surgery and 6 subsequent brain surgeries and two gamma knife procedures) and was admitted to University Hospital for Infectious Diseases “Fran Mihaljević”, Zagreb, Croatia in 2014 for pre-operative medical treatment of brain hydatidosis in the right parietal region. We aimed to attain a high cyst albendazole sulphoxide (ASO) concentration in order to achieve a more pronounced protoscolex inactivation and a high serum ASO concentration (reflecting the tissue concentrations) to reduce the risk of disease recurrence. The patient was treated with a higher dose of albendazole (15 mg/kg/day for 4 wk) that we had found effective in patients with liver hydatidosis, and combined with praziquantel over the last 14 d at a dose that is typically used to treat neurocysticercosis with an intention to improve ASO bioavailability. Neither serum nor cerebrospinal fluid concentrations on day 10 apparently differed from those on day 24 indicating a lack of an effect of praziquantel on ASO bioavailability. Intra-cystic ASO concentration was below the lower limit of quantification, but above the limit of detection. After the 7^th episode of the disease and combined albendazole-praziquantel and surgery treatment, the patient achieved a 3-year remission. With the apparent lack of a meaningful pharmacokinetic praziquantel-albendazole interaction, this is most likely ascribable to the use of a higher albendazole dose than previously.

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