Background: Leishmaniasis is important vector-borne parasitic disease worldwide, caused by the genus Leishmania. The objective of the current study was to identify genetic polymorphism in L. major, one of the species causing cutaneous leishmaniasis (CL), isolated from southeastern Iran, using Permissively Primed Intergenic Polymorphic-Polymerase Chain Reaction (PPIP-PCR) method.

Methods: Overall, 340 patients with suspected CL were examined. They referred to the Central Laboratory in Chabahar, Iran during Apr 2013 to Feb 2014. Microscopic examination of Giemsa-stained slides from lesions as well as aspirates cultured in Novy- Mac Neal-Nicolle (NNN) Media was employed in order to diagnose CL in these patients. Our analyses detected 86 suspected subjects as having CL from which 35 isolates were cultured successfully. PPIP-PCR method was performed on extracted genomic DNA from selected isolates in order to determine the genetic polymorphism among L. major isolates.

Results: The electrophoresis patterns demonstrated two genetic profiles including A or A1 patterns between all samples tested. Frequency of A and A1 sub-types were 33 (94.3%) and two (5.7%), respectively.

Conclusion: Both host and parasite factors may contribute to the clinical profile of human leishmaniasis in the endemic foci of the disease. Here we showed that genetic variations pertaining to the Leishmania parasites might determine, in part, the clinical outcomes of human leishmaniasis. 

Background: The present study was carried out to detect the infection of larval stages of Trichobilharzia species in the snail Lymnaea auricularia in northwestern Iran based on DNA analysis.

Methods: A total number of 320 snails of L. auricularia were sampled from four water-bodies located in the suburb of Urmia City, North West Iran, during May to November 2011. The snails were first microscopically inspected for the infection with larval stages of trematodes. Genomic DNA was extracted from the snails and PCR was performed to amplify a fragment of the ribosomal DNA of Trichobilharzia species in the infected snails.

Results: Microscopic examinations indicated that 11.25% (36 out of 320) of the snails were infected with larval stages of trematodes, while the PCR patterns showed a much higher infection rate (31.25%, 100/320). According to the PCR, the infections were caused by the larval stages of T. szidati (21.56%, 69/320) and T. franki (9.69%, 31/320) or both of them (8.44%, 27/320). The infected snails were observed in three out of the four studied sites. The highest infection rate in a single site was 50% (25/50). Only 7.81% (25 out of 320) of the infected snails were from the plain areas, while the remaining was from high altitudes.

Background: We aimed to reveal the role of CD11b and hypoxia-inducible factors-1alpha (HIF-1α) expressions on monocytes and alveolar macrophages of lung tissue, and the levels of serum surfactant protein-D (SP-D) in severe malaria-associated acute lung injury (ALI).

Methods: The C57BL/6 mice were divided into control group, renal malaria group (inoculated with 10⁶ Plasmodium berghei ANKA), and cerebral malaria group (inoculated with 10⁷ P. berghei ANKA). The expressions of CD11b and HIF-1α in lung tissue were observed by immunohistochemistry, and serum SP-D levels were measured by ELISA. This study was conducted from June 2014 to February 2015 in the Laboratory of Parasitology, Faculty of Medicine, Universitas Brawijaya, Malang.

Results: The CD11b expression on pulmonary tissue of renal and cerebral malaria mice were significantly higher than control mice (P=0.002; P=0.002), as well as the HIF-1α expression on pulmonary tissue (P=0.002; P=0.002). The level of serum SP-D in renal malaria and cerebral malaria mice were significantly higher than control mice (P=0.002; P=0.002). We found a strong correlation between the expression of CD11b and HIF-1α in lung tissue (r=0.937, P=0.000), as well as between CD11b expression and serum SP-D levels (r=0.907, P=0.000) and between HIF-1α expression and serum SP-D levels (r=0.913, P=0.000).

Background: Toxoplasma gondii is an obligate, intracellular protozoon that develops its sexual stage in cat’s intestinal epithelial cells as definitive host and develops its asexual stage in different tissues of a wide range hosts called intermediate host. The protozoon is a food-borne and worldwide parasite that can cause serious complications such as abortion in pregnant women, encephalitis, and ocular toxoplasmosis. The present study aimed to genotype T. gondii strains isolated from patients with toxoplasmic retinochoroiditis.

Methods: Fifty-two blood samples were taken from patients with ocular toxoplasmosis, from July 2013 to July 2014. The specimens were collected from three ophthalmological hospitals of Tehran, Iran. After that, DNA extraction was performed using kit on separated buffy coats of serologically positive blood samples. Then PCR was done in GRA6 gene. For digestion of products, MseI endonuclease was used. Finally, some of the PCR products were sequenced.

Results: All of 52 samples were found positive by serological and PCR-RFLP methods and all of isolated strains belong to type III genotype. Type III genotype has the highest prevalence in Iranian ocular toxoplasmic patients.

Background: The present study was conducted to investigate the optimized timing for macrophages induction and nitric oxide (NO) production against invading Leishmania parasite.

Methods: The present study examined the murine macrophage cell line, B10R, in three different states. In the first state, the cells were first infected with L. major and then treated with IFN-g and LPS as stimulants. In the second state, the cells were infected after stimulation with IFN-g and LPS. In the third state, the cells were only exposed to stimulants as controls. In all the three states, cell culture supernatants were collected at three points in time (6, 24 and 48 h) and the amount of NO production was measured using Griess assay.

Results: The treatment of macrophages with inducers prior to infection with stationary phase parasite led to the secretion of significant amounts of NO, particularly at early time points quit contrary to the cells infected with parasites prior to induction. The amount of NO produced by cells induced after infection was detected significantly lower.

Conclusion: The induction of macrophages prior to infection with parasites leads to the production and secretion of greater amounts of NO, resulting in an increased ability to suppress and inhibit parasite proliferation even in the early stages of infection.

Background: The present study aimed to survey antileishmanial activity of methanolic Holothuria leucospilota extract against Leishmania major promastigotes in vitro.

Methods: Promastigotes were cultured in RPMI 1640 and after reaching the stationary phase, the study was conducted with different concentrations of the extract. Afterwards, MTT colorimetric assay for the obtaining of 50% inhibitory concentration (IC₅₀) was utilized. Furthermore, in order to determine the possible induction of apoptosis in L. major promastigotes, flow cytometry and DNA fragmentation methods were employed using annexin-V FLUOS staining kit and DNA ladder kit, respectively.

Results: The IC₅₀ value of H. leucospilota extract at three time points of 24, 48, and 72 h was estimated 2000, 300 and 85 µg/ml, respectively. In addition, the extract revealed a dose and time-dependent antileishmanial activity. Furthermore, various characteristics of apoptosis appeared after L. major promastigotes treatment, which included cell shrinkage, formation of apoptotic bodies, blebbing of the cell membrane, and externalization of phosphatidylserine, although no laddering pattern was observed.

Conclusion: The methanolic extract of H. leucospilota possesses lethal effect on L. major promastigotes and induces the apoptosis in parasites. Further studies are required to address the apoptosis mechanism in vivo.

Background: This study was conducted to collect informative data on the parasitic infection of wild rodents, emphasizing on finding parasites, which have medical importance to human.

Methods:  During 2012-2014, a total number of 91 wild rodents were captured from rural areas of Turkmen Sahra, Golestan Province, using handmade traps. Animals were anesthetized, surveyed for any ectoparasite and then their carcasses were carefully dissected for examination of endoparsites.

Results: Four species of rodents including Mus musculus (52.75%), Rattus norvegicus (38.46%), Rhombomys opimus (4.40%) and Meriones libycus (4.40%) were captured. Parasitic infestation was detected in 38.5% of sampled rodents. Parasite infestation rates of sampled rodents was Hymenolepis diminuta = 7.7%, Cryptosporidium spp = 6.6%, Trichuris spp.= 5.5%, Cysticercus fasciolaris = 2.20%, Angiostrongylus spp.= 2.20%, Capillaria sp.= 1.09%, Rhipicephalus spp. = 8.70%, Nosopsyllus fasciatus = 1.09%, and Laelaps nuttalli = 3.29%. Among 10 genera/species of identified parasites, at least 8 of them were zoonotic with public health importance. L. nuttalli and N. fasciatus were the only two non-zoonotic detected parasites in this survey.

Conclusion: Harboring a wide variety of zoonotic parasites in sampled wild rodents particularly when they live nearby villages, represents a potential risk to native inhabitants. Hence, controlling rodents’ population in residential regions and improving awareness of local people about the risk of disease transmission through rodents seems to be entirely necessary.

Background: We investigated the variations of heamathologic and biochemical parameters with different parasitic burdens and the correlation between fecal egg counts and fluke number in livers of sheep naturally infected with Dicrocoelium dendriticum.

Methods: In order to detect excretion of D. dendriticum eggs, fecal samples from 120 sheep of different age groups were collected individually at slaughterhouse in Neyshabur County in Razavi Khorasan Province during December 2013 - October 2014. Hematologic and blood biochemical values in 120 sheep naturally infected with D. dendriticum were compared with equivalent values in 120 parasite-free sheep from an organically farmed flock. Investigated animals were kept in outdoor system, on pastures covered with swamps, which remain flooded after rainy season.

Results: Total numbers of flukes in livers obtained from infected sheep at autopsy varied between 10 and 18,500. A positive relationship was noted (r= 0.94, P < 0.001) between the number of eggs excreted by each sheep and that of total fluke counts in the liver. Significantly, lower red blood cell, packed cell volume, neutrophil and albumin were observed in parasitized sheep, when compared to the healthy animals. White blood cells,, lymphocyte and aspartate aminotransferase values were significantly higher in parasitized sheep. No significant correlation was detected between the hematologic and biochemical parameters and the number of D. dendriticum in the liver.

Background: This study aimed to investigate the epidemiological and clinical aspects of patients with hydatid cyst during 2011 to 2014.

Methods: This cross-sectional study was conducted in Khorasan Razavi Province, the Northeast of Iran, from 2011 to 2014. The study population was all cases with hydatid cyst who diagnosed in governmental and private laboratories, hospitals and health centers (HC) in Khorasan Razavi Province during 2011-14.

Results: The prevalence rate of hydatidosis was 1.44 per 100000 individuals. Of 357 cases, 54.9% were women, 40.3% rural, 45.8% housewives, and 3.4% were Afghan. The mean age of women was higher than that of men (39.13±18.9 compared to 34.7±17.9 yr, respectively, P-value=0.025). The highest proportion of cases (39.2%) was in the age group of 21-40 yr old. Abdominal pain was reported in 42.3% of cases. Liver involvement was the most common localization of hydatid cyst reported in 59.4% of patients, and 8.4% had multiple organ involvement. The common diagnosis methods of the disease were radiology (42.3%) followed by CT scan (37.8%). 45.9% of patients had domestic dog and hygiene principles of washing the vegetables was adhered by 6.7% of patients.

Background: Aurones are naturally occurring compounds that belong to flavenoids family and have antiplasmodial effects. This study investigated some new aurones derivatives against chloroquine sensitive Plasmodium falciparum. Here we report the synthesis, in vitro antiplasmodial activity and cytotoxic evaluation of 11 compound from derivatives of (Z)-2- benzylidene-4, 6-dimethoxybenzofuran-3(2H)-one.

Methods: The cytotoxic evaluations of active compounds were performed with MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyltetrazolium bromide) assay on human breast cancer cell lines; MCF7 and T47D.

Results: From 11 compounds M3, M6 and M7 compounds showed good antiplasmodial effect against chloroquine-sensitive 3D strain of P. falciparum with IC₅₀ (50% inhibitory concentration) values of 7.82, 7.27 and 2.3 µM respectively. No noticeable toxicity was‌ observed with these compounds when tested against tested cell lines.

Background: Wild boars, Sus scrofa, of wide distribution considered as a potential source of zoonotic parasites. The current study aimed to assess the prevalence of helminth infections in wild boars in the Persian Gulf coastal area (Bushehr Province), Southwestern Iran.

Methods: Twenty-five wild boars, including 11 males and 14 females, were collected during a course of vertebrate pest control in the Bushehr Province, southwestern Iran in 2013. The specimen were immediately dissected and carefully searched for the parasites. During necropsy, each organ was examined macroscopically for presence of any helminthic agents. Tissue samples were taken from each organ. Moreover, samples were taken from the content of digestive system. Blood samples were also collected from each boar. All the samples were evaluated for helminth infections by parasitological methods.

Results: Twenty-two (88%) of the wild boars were infected with at least one helminth. Out of 25 wild boars, 1 (4%) were infected with Cysticercus tenuicollis, the larval stage of Taenia hydatigena, 13 (52%) with Macracanthorhynchus hirudinaceus, 17 (68%) with Metastrongylus spp, and 20 (80%) with Ascarops spp. Hydatid cyst was detected in the lung of one of the wild boars. No Trichinella spp. larvae were detected in any of the tissues of the animals when evaluated by artificial digestion method. In addition, no contamination with microfilaria was detected in any of animals when the blood samples were tested with Knott’s method.

Conclusion: Wild boars are contaminated by some helminthes including zoonotic ones. These animals could be involved in the epidemiology of zoonotic helminth by acting as reservoir hosts. This in turn may bring potential risk for locals and residents of the Bushehr Province, Southwestern Iran.

Background: This investigation aimed to evaluate the in vitro scolicidal effects of Pistacia khinjuk methanolic extract against protoscoleces of hydatid cysts and its acute toxicity in mice NMRI model.

Methods: Protoscoleces were aseptically extracted from sheep livers having hydatid cysts. Various concentrations of the essential oil (12.5- 100 mg/mL) were used for 10 to 60 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). Twenty-four male NMRI mice were used to assess the acute toxicity of P. khinjuk.

Results: P. khinjuk extract at the concentrations of 100 mg/mL after 10 min of exposure killed 100% of protoscoleces. Similarly, the mean of mortality rate of protoscoleces after 20 min of exposure to the concentration of 50 mg/mL was 100%. The LD₅₀ of the intraperitoneal injection of the P. khinjuk methanolic extract was 2.8 g/kg and the maximum non-fatal dose was 1.7 g/kg.

Background: The study evaluated qualitative PCR, primers 121-122 as a tool to follow up evolution parasite load of Trypanosoma cruzi.

Methods: The study was conducted at the State University of Maringa, in 2015. Step 1, dilutions 1/10 were performed from T. cruzi-Y strain to obtain preparations of 50,000-0.05 parasites/mL from which DNA were extracted, quantified, and amplified. Step 2, the extracted DNA in the dilutions 5-0.05 parasites/mL was re-diluted 1/10, 1/100, 1/1000, quantified, and amplified. Polyacrylamide gels were photographed and thicknesses of the 330 bp kDNA fragments were measured.

Results: Step 1, in the dilutions 50,000-50 parasites/mL kDNA fragments had same thickness and, dilutions 5-0.05 parasites/mL showed progressive decrease in thicknesses and staining intensity of the 330 bp fragments. Step 2, demonstrated that dilutions of five (re-dilutions 1/10 and 1/100) and 0.5 (1/10) parasites/mL produced similar thicknesses of the 330 bp fragments obtained in Step 1. However, very dilute DNA samples make difficult to reproduce the fragments thicknesses.

Background: Weak efficacy of different fasciolicidal compounds used for treatment of paramphistomosis has drawn the attention of many authors to alternative drugs. The purpose of this study was to assess, for the first time, the effect of the methanolic extract of Balanites aegyptiaca fruits (BAE) on adult Paramphistomum microbothrium.

Methods: The effect of BAE on adult P. microbothrium after 24 h incubating the parasites in RPMI 1640 culture medium containing 10, 50, 100 and 200 µg/ml BAE was determined by light and scanning electron microscopic studies.

Results: Differences in response to BAE action were concentration dependent.The major target organ that was highly affected was the tegument. Maximum anthelmintic activity was found with a dose of 200 µg/ml BAE, at which distinct damage to the whole body surface of the trematodes was very much distinct. Shape and structure of both suckers were deformed due to BAE. This damage would undoubtedly disrupt many of the physiological processes associated with the tegument. Besides, the damage of the tegumental folds of the acetabular region might disrupt its function in drawing the rumen wall tissue of the host into the acetabular cavity.

Conclusion: the use of methanolic extract of B. aegyptiaca fruits offers a new dimension and potential for control of such a neglected infectious disease in ruminants, at a time when paramphistomosis has emerged as an important cause of productivity loss.

Background: Hydatid cysts are encountered frequently in regions endemic with livestock. The basic treatment for a hydatid cyst is total surgical removal of the cyst and its inner contents. Hypertonic NaCl or diluted betadine solution are used as germicidal agents for most hydatid surgeries. However, the germicidal efficacy of the Ankaferd Blood Stopper® (ABS) has not been investigated. Thus, we compared the efficacy of ABS for hydatid cysts with that of other germicidal agents.

Methods: Lung and liver tissues containing hydatid cyst liquid were collected from slaughterhouses. Six samples of each cyst were randomly allocated into different groups as follows: 20% hypertonic NaCl, betadine solution, ABS, 20% liquefied Andazole solution, 0.1% eosin, and distilled water. All groups were examined microscopically at 5, 10, and 15 min after treatment began to determine protoscolece viability rates.

Results: The most efficacious germicidal agent at 5 min was ABS, and betadine and hypertonic NaCl had similar efficacies. Betadine, ABS, and hypertonic NaCl showed similar efficacies at 15 min. 

Conclusion: ABS was an effective germicidal agent to treat hydatid cysts.

Strongyloidiasis is a frequent misdiagnosed parasitic infection in the world that caused by Strongyloides stercoralis. In Iran, the disease is predominantly reported from warm and humid climate provinces. The patient was a 54-yr-old man, originated from Khuzestan Province with a history of pemphigus and diabetes that was treated with high-dose of corticosteroid drugs before admission in a non-private hospital in Shiraz, Iran in 2014. After different primary diagnosis and administrating of several drugs, endoscopy and histopatholgical biopsy revealed a massive S. stercoralis infection in the duodenal mucosa and gastric wall. In spite treating with anti-helminthic drugs in the last days, due to using different steroid drugs, clinical manifestations of the patient were exacerbated and he was expired on the seventeenth day due to severe dyspnea. Physicians’ awareness and using various diagnosis methods like serology, endoscopy, and biopsy should be considered in the endemic areas. In suspicious cases, anthelmintic drugs should be started before the initiation of immunosuppressive therapy.

Brain hydatid cyst constitutes one of the leading causes of cranial mass in the endemic region, especially among the children. Symptoms of the disease are associated with rising of intracranial pressure and focal neurological deficit. Surgical excision of the cyst is essential to treatment. Here we present a 19 yr-old male livings in Ardabil, northwest of Iran, referred to Neurosurgery Clinic in Khanevadeh University Hospital, Tehran, Iran with a large brain hydatid cyst and atypical symptoms. He was treated by surgical excision of the cyst without any complication. There were not any clinical or radiological signs of recurrence after one-year follow-up.