Iranian Journal of Parasitology
Volume:9 Issue: 2, Apr-Jun 2014

Species of Microsporidia have been known as opportunistic obli­gate intracellular parasites particularly in immunocompromised patients. Enterocyto­zoon bieneusi is one of most prevalent intestinal microsporida parasites in HIV+/AIDS patients. In this study, intestinal microsporidia infection was deter­mined in HIV+/AIDS patients using microscopic and molecular methods. Stool samples were collected from HIV+/AIDS patients during 12 months. All of the stool specimens washed with PBS (pH: 7.5). Slim slides were prepared from each sample and were examined using light microscope with 1000X magnification. DNA extraction carried out in microscopic positive samples. DNA amplification and genus/species identification also performed by Nested-PCR and sequencing techniques. From 81 stool samples, 25 were infected with microsporidia species and E. bieneusi were identified in all of positive samples. No Encephalitozoon spp. was identified in 81 collected samples using specific primers. E. bieneusi is the most prevalent intestinal microsporidia in immunocompromised patients of Iran. On the other hand, Nested-PCR using spe­cific primers for ssu rRNA gene is an appropriate molecular method for identifica­tion of E. bieneusi.

Clinical manifestations of Strongyloides stercoralis are variable from asymptomatic to hyperinfection and devastating disseminated infections. Hereby, clinical characteristics of a large series of Iranian strongyloidiasis indigenous cases are described. The records of people referred to the Helminthological Diagnostic Labora­tory of School of Public Health, Tehran University of Medical Sciences and School of Medicine, Gilan University of Medical Sciences, during 2009-2013 were reviewed. For those patients that were infected with S. stercoralis and their clinical manifestations and demographic data were available (70 cases) a checklist was pre­pared and data analyzed. Forty-three patients (61.4%) were male and 27 (38.6%) female. Gastrointesti­nal, cutaneous and pulmonary symptoms were present in 71.4%, 25.7%, and 15.7% of patients, respectively. None of them had larva currens erup­tion. Eosinophilia was the most prevalent reason for suspicious on S. stercoralis, but the mean was lower in elderly patients. Hyperinfection were recorded in 8 patients (11.4%), and 2 cases had disseminated infection. Eosinophilia is common both in asymptomatic and symptomatic cases of strongyloidiasis, but the mean tend to lower with increase in age.

To provide a point of reference to study the epidemiology and clini­cal expression of canine babesiosis in China. A total of 30 dogs infected with canine babesiosis were evaluated by mean of clinical history, physical examination, hematological, restriction fragment length polymorphism of PCR products (PCR-RFLP) and sequencing analysis. The most prevalent clinical abnormalities were lethargy (100%), anorexia (100%), pale or icteric mucous membranes (80%), fever (70%) and dark urine (70%). Hematology parameters revealed that anemia and thrombocytopenia were the major abnormalities in blood of dogs infected with canine babesia. The results of PCR-RFLP and sequencing analysis indicated that B. gibsoni was the main species responsible for canine babesiosis cases at the time of the study in Nanjing, China. The results provide valuable information for better understanding of the epidemiology of canine babesiosis in China.

Among the infectious organisms of parasitic origin, gastrointestinal nema­todes are very important as they have been reported worldwide. The main aim of the present research study to highlight the annual epidemiological contributing factors associated with the prevalence of gastrointestinal nematodes and their control in sheep. A total 1200 faecal samples (100 per month) were collected from farmers holding Balochi-sheep (either sexes, 1-5 years old) during January-December 2012 and analyzed to determine the prevalence of nematodes based on microscopy and ELISA based diagnostic assay. Therapeutic efficacies of different synthetic and herbal medicines against these nematodes were assessed by field trials. Results showed that 23.92% Balochi-sheep were infected with nematodes. Five nematodes infections were recorded with highest prevalence of Haemonchus (7.75%) fol­lowed by Nematodirus (7.58%), Strongyloides (4.42%), Trichostrongylus (2.33%) and Trichuris (1.83%). The younger and older ewes (one and five years) presented higher nematodes prevalence with peak during March/April and August/September. Haemonchus and Trichu­ris positive samples based on coprological examination were also showed 92-100% positive sensitivity for these nematodes by the ELISA. Sheep treated with Ivermectin showed higher reduction (97.76%) in nematode egg counts followed by Atreefal deedan (96.42%) and Oxfendazole (95.44%), respectively. The gastro-intestinal nematodes are prevalent in all age and either sex of Balochi-sheep with peak during summer. The ELISA based diagnosis is more accurte. The synthetic and herbal products are very effective against sheep nematodes.

Toxoplasma gondii is one of the most prevalent parasites of human and warm- blooded animals. Toxoplasmosis is important especially in two groups: pregnant women and immunocompromised patients. If women acquire the primary infection during the pregnancy, it would be life threatening or re­mains severe disorders for the fetus. This study was performed to evaluate the seroprevalence of T. gondii infection in pregnant women referred to Health Cen­ter in Gorgan City, Golestan Province, northern Iran. Serum samples were collected from pregnant women referred to Health Center in Gorgan City, south eastern Caspian Sea. Anti- Toxoplasma IgG and IgM antibodies were determined by commercially ELISA kits and the rela­tion of infection with socio-demographic and risk factors such as age, education, occupation, cat ownership, soil contact and some other factors was studied. From 555 tested sera of pregnant women referred to Health Center in Gorgan, 39.8% had IgG antibodies against T. gondii and 3.4% were positive for IgM antibodies. A significant correlation was seen between T. gondii infection with age and soil contact. About 60% of pregnant women in Gorgan City are seronegative against T. gondii, so they should considered as at risk persons.

In this paper, paleoparasitological findings from rodent excrements ob­tained from Chehrabad Salt Mine archeological site located in northwest of Iran are demonstrated and discussed. Chehrabad Salt Mine archeological site located in northwest of Iran, dated to the Achaemenid (mid 1st mill. BCE) and to Sassanid (3rd cent. - 7th cent. CE) period, is a unique study area to investigate parasites in the past millenniums in Iran. Rodent copro­lites obtained from this archeological site were thoroughly analyzed for parasite eggs using TSP re-hydration technique. Specimen analyzed were attributed to juvenile and adult rats based on their apparent morphology comparing with the modern dried pellets of Muridea family. Hel­minth eggs retrieved from two positive pellets were identified as Trichosomoides crassi­cauda, Syphacia sp. and Trichuris sp. The present paper discusses the first paleoparasitological findings of rodent gastrointestinal helminthes in Iran along with possible favorite items to rats in ancient Chehrabad Salt Mine.

The free-living amoebae Acanthamoeba spp. have been recognized as etiologic agents of amoebic encephalitis, keratitis, otitis, lung lesions and other skin infections mainly in immuno-compromised individuals. The purpose of this study is to detect the presence of Acanthamoeba in swimming pools in Egypt using a polymerase chain reaction (PCR) method. Water samples were collected from 10 different swimming pools in Cairo, Egypt. Samples were cultured on non-nutrient agar for the detection of Acan­thamoeba isolates that were confirmed by PCR amplification using genus spe­cific primers. The molecularly confirmed Acanthamoeba isolates were morphologi­cally identified to the species level. Members of genus Acanthamoeba were detected in 49.2% of the examined swimming-pool water samples. Morphologically, six Acanthamoeba species were iso­lated from the examined swimming pool water namely A. polyphaga, A.castellanii, A. rhysodes, A. mauritaniensis, A. royreba and A. triangularis. All the identified species of Acanthamoeba were molecularly confirmed to be related to the genus Acanthamoeba. The isolated species of Acanthamoeba could provoke variable degrees of infections to the swimmers. The culture method is cheaper and easier than PCR techniques that are faster for the detection of free-living amoebae

The aim of this study was to evaluate the protein spots of excre­tory - secretory products of Fasciola hepatica using two dimension electrophoresis method in the presence and absence of triclabendazole drug which can be consid­ered to detect the target protein of the drug. F. hepatica parasites were collected from infected cattle livers, divided in two groups and cultivated in RPMI 1640 medium. First group was treated with triclabendazole (TCBZ) and second group considered as control. The excretory-secretory (ES) products of each group were separated and total protein deter­mined by Bradford method. To provide proteome spots, the ES proteins were precipitated and two dimension electrophoresis (2-DE) gel prepared. Protein amounts of two groups were compared using the statistical t-test and protein spots from 2-DE in test and control groups were also statistically analyzed. The protein spots of gels were identified by using protein database. The t-test showed a significant increase of total proteins in treated group (P<0.5). The protein spots count in the control group was less than test group however statistically not significant (p>0.05). Cathepsin L- protein (MW 36.7 pH 5.34), 14-3-3 epsilon 2 isoform (MW 28.2 pH 5.36), Cathepsin L1D (MW 36.5 pH 5.8) and Cathepsin L1D (MW 36.6 pH 6.26) were identified in test group. It seems that, these results can be considered to determine the pro­teins which the drug acts as a target on them.

Parasitic nematodes cause animal and human diseases of major socio-economic importance worldwide. The suppression of parasite development at particular developmental stages could provide an alternative approach for nema­tode control. In this study, Ascaris suum was used as a model system in the study of the differentially expressed genes in the infective L3 stage. The gene (07E12) was screened and identified from the subtractive cDNA library for the infective larvae of Ascaris suum using real-time quantitative PCR. Then, the full-length cDNA of 07E12 was characterized by 3′ and 5′ rapid amplification of cDNA ends (RACE). The characteristics of the gene were further analyzed using bioinformatic analyses. The results showed that the gene 07E12 was differentially expressed in the third-stage larvae of A. suum and its expression level in the infective larvae was much higher than in other stages. It was shown that the gene 07E12 had 99% iden­tity with the corresponding sequences of the A. suum whole genome shotgun se­quence containing the homologous sequences with conserved sequences of Neuropep­tide-Like Protein family member. Likewise, by performing BLASTN and BLASTP searches in the GenBank™, it was shown that this gene had 99 % identity with A. suum cre-nlp-2 protein. This gene 07E12 which is differentially expressed in the third-stage larvae of A. suum may encode a neuropeptide-like protein family member, a very important molecule in the process of infecting a host.

Cystic echinococcosis caused by the metacestode of Echinococcus granulosus is a major problem in both humans and domestic animals health. There­fore, a standardized and approachable diagnostic tool (rapid tests) for the serodiagno­sis of cystic echinococcosis (CE) is still needed. In the present work, antigen B labeled with gold nanoparticles was used to detect antibodies against hydatid cyst disease. The prepared antigen B was ana­lyzed by SDS-PAGE. Tetra chloroauric acid (HAuCl4) was used to produce colloi­dal gold and antigen B labeled by gold nanoparticles, then it was tested by using rabbits antisera and sera from naturally infected sheep. The labeled antigen B was evaluated using Dot-immunogold staining (Dot-IGS) method. Electrophoretic pattern of hydatid cyst fluid showed the quality of bands in the condensed fluid is better than crude fluid. SDS-PAGE analysis cyst fluid and antigen B revealed three specific protein bands that were detected at molecular weights of 24, 30 and 40 kDa that all are the subunits of antigen B. Evaluation of antigen B labeled by gold nanoparticles by using Dot-IGS technique showed 1/1 and 1/50 dilutions in comparison with another has the best immunoreaction. In this method, nanoparticles produced a typical purple color, when they binded to the strip at the site of immunoreaction. Therefore, using gold nanoparticles is a good candidate for detection of helminthiasis, also as selective tools of early detection, simple and cost-effective, regardless of specific skills and equipment with optimal durability.

Cryptosporidiosis is a common coccidian parasite infection in patients with diarrhea that has worldwide distribution especially in developed countries. There­fore, the aim of this study was to determine the occurrence of Cryptosporidium infec­tion in patients with gastroenteritis admitted to hospitals of Mazandaran University of Medical Sciences by parasitological and molecular methods in Sari, Iran. Stool samples were collected from 348 patients with gastroenteritis admit­ted to the hospitals of Medical University in the Sari and Ghaemshahr cities in Mazanda­ran Province, Northern Iran in 2010-2011. Oocysts of Cryptosporidium identi­fied using Formalin-Ether concentration method and stained by Aacid-fast staining (AFS) and Auramine phenol fluorescence (APF). Genomic DAN extracted from micro­scopically positive samples and nested PCR -RFLP by using SSU rRNA that identifies of the species of cryptosporidium. In 348 patients with gastroenteritis, the most clinical symptoms were diar­rhea, nausea, vomiting, dehydration, fever and weight loss. 2.3% (8 cases) of diarrheal samples tested by both microscopy and molecular methods were positive for the pres­ence of cryptosporidium. Nested PCR products yielded unique bands of 846 bp, corre­spond to cryptosporidium. Species diagnosis carried out by digesting the second­ary PCR product with SspI restriction enzyme, which noted 3 clearly bands of 449, 254, and 108 bp correspond to Cryptosporidium spp. The results of present study on Cryptosporidium spp. in this area can make a background data for control programs and further molecular analyses. Thus, further work needs to determine the origin of Cryptosporidium species in this area.

Hydatidosis, as the most important zoonotic parasitic disease in Iran, has posed many health and economic losses. This study was conducted to investigate the demographic characteristics of hydatid cyst surgeries in hospitals of East Azerbaijan Province, Northwest of Iran. Demographic characteristics of all patients with hydatid cyst surgery in hospitals of the province, during 2009-2011 were gathered including age, gen­der, occupation, number and location of the cyst, clinical symptoms, place of residence and history of contact with dog. They were extracted from reports of health center and were analyzed using STATA 11 software. Out of 52 hydatid cyst surgeries, 27 cases were females. Mean age of patients was 38.3 yr. Liver was reported as the most involved organ. The most clinical symptoms were abdominal and liver pain. Housewives comprised the most victims of the disease. Forty seven percent of patients had one cyst and 59% had the history of contact with dog. The majority of the patients were liv­ing in rural areas. Due to the high costs of diagnosis and treatment of hydatidosis, collecting data on the prevalence and transmission of the disease as well as on vulnerable groups seems to be essential as the first step in controlling and prevent­ing the disease.

Cutaneous leishmaniasis (CL) is endemic in Southeastern Anatolia, mainly in Sanliurfa and Hatay provinces, and the causative agents are mostly Leishma­nia tropica and less frequently L. infantum. Here, we report the first MALDI-TOF analyses of Leishmania promastigotes obtained from the cultures of two CL cases from Osmaniye and Hatay provinces who were initially diagnosed by microscopy, culture and identified as L. infantum with Real-Time PCR (RT-PCR). Samples obtained from the skin lesions of patients were initially stained with Giemsa and cultivated in NNN medium. Examination of the smears and cultures revealed Leishmania amastigotes and promastigotes, respectively. The promastigotes (MHOM/TR/2012/CBU15 and MHOM/TR/2012/MK05) ob­tained from the cultures of both patients were used for RT-PCR targeting the ITS-1 region in the SSU of rRNA. The reference strains of four Leishmania spe­cies (L. infantum, L. donovani, L. tropica and L. major) were initially assessed with MALDI-TOF and their data were added to MALDI-TOF Biotyper Library. Both RT-PCR and MALDI-TOF analyses indicated that the causative agent in both patient samples was L. infantum. Despite disadvantages such as requirement of culture fluid with noth­ing but promastigotes and high cost, MALDI-TOF analysis may be a fast, sensitive and specific diagnostic tool in especially large-scale research studies, where the cost declines, relatively.

In this experiment, abdominal cavity of 518 Iranian Sistani cattle and 498 Brahman cattle were inspected for the presence of Setaria spp. from April 2012 - May 2013. The species were determined by microscopic examination of the morphologi­cal characteristics of the anterior and posterior parts of the parasites and authentic guidelines. The overall prevalence of Setaria spp. was 28.6% and 36.5%, respectively and this difference was significant (P<0.05). Out of 148 Sistani cattle which were infected with Setaria, 51(34.4%) were infected with S. digitata, 31 (20.9%) were in­fected with S. labiatopapillosa, 65 (43.9%) showed mixed infection of S. digitata and S. labiatopapillosa and one case (0.6%) was infected with mixed infection of S. labiatopapil­losa, S. digitata and S. marshalli. These values were 87 (47.8%), 27 (14.8%), 67 (36.8%) and 1 (0.5%) for 182 infected Brahaman cows, respectively. The propor­tion of infected cattle in spring and summer was greater than cooler season (autumn and winter) significantly (P<0.001). The prevalence of infection with Setaria in 2-3 years old Sistatni cattle (42.2 %) was greater than other age categories (P<0.05). Furthermore, the infection rate between males (25.5%) and females (37.3%) Iranian Sistani cattle showed significant difference (P =0.009). It is important to point out the presence of cerebrospinal setariosis, namely in sheep, goats and horses in the investigated area.

Naegleria is a free-living amoeba, and pathogenic Naegleria may pose a health risk to people exposed to recreational water. Our objective in this study was to determine if there are pathogenic amoebae in environmental water samples from Changchun, Northeastern China. During July to September 2012, a total of 70 water samples were collected from Changchun, Northeastern China, and Naegleria was enriched by in vitro culture and detected by PCR using Naegleria genus-specific primers. Result­ing PCR products were sequenced and phylogenetically analyzed to iden­tify Naegleria species. Naegleria was detected in 65 (92.9%) of 70 water samples. DNA se­quence and phylogenetic analyses based on the internal transcribed spacer (ITS) rDNA sequences revealed four Naegleria species, including N. pagei (n = 24) and N. Australiensis (n = 18), N. clarki (n = 13) and N. gruberi (n = 10), in which N. australiensis is pathogenic to mice. But the pathogenic species N. fowleri was not detected. This is the first report on Naegleria species in Northeastern China, showing that almost all environmental water samples were contaminated with Naegleria, including N. pagei, N. Australiensis, N. clarki and N. gruberi, which should be considered a potential public health threat.

Angiostrongylus cantonensis is a zoonotic public health concern that causes human severe eosinophilic meningitis in Southeast Asia and China. As a medically important intermediate host of A. cantonensis, Bellamya lithophaga (Gastrop­oda: Viviparidae) is often confused with other morphologically similar sibling spe­cies of genus Bellamya, such as B. aeruginosa and B. purificata in the past. Hence, the aim of the present study was to investigate evidences to discriminate these equivo­cal Bellamya species. This study was carried out by getting Bellamya snail samples from Fujian Province in the South-East of China. The snail morphological features，breeding grounds and phylogenetic relationship according to mitochondrial cytochrome c oxidase subunit I (COI) gene marker were analyzed. Based on external morphology, radular shape and cusp formula, as well as major breeding environment, B. lithophaga could be distinguished from B. aeruginosa, B. purificata. The phylogenetic tree also unconfirmed that B. lithophaga belongs to a different genetic clade from other morphologically similar species. Our findings demonstrate the significant differences in B. lithophaga and other sibling species, which supports the traditional species delimitation in the genus Bellamya.

Main objective of this study was to investigate the invasive activity of the liver fluke, Pseudamphistomom truncatum against the Caspian seal (Pusa caspica) and was exemplified at the gross, light microscopy (LM) and electron microscopy (EM) levels. The study was done on a freshly dead Caspian Seal in the southern coast of Caspian Sea. The checked Caspian seal probably being died of canine distemper virus and was found host to numerous parasites of four helminth species. P. truncatum caused edematous foci on the surface of the liver with promi­nent fluid accumulation. Sections of the liver viewed with LM had multiple necrotic areas with extensive hemorrhaging and disorganized hepatic lobules. Granulocytes and invasion of connective tissue were prominent. Whole worms were visible with invasive pathways through the host tissue. Damage to both hepatic ducts and blood vessels were prominent. At the EM level, organelles within the impacted hepato­cytes were disorganized as exemplified by the cristae of the mitochondria and the endoplasmic reticulum. Parasite eggs were scattered throughout the tissue. It was shown that this trematode can be very pathogenic to Caspian Seal and as this only mammal of Caspian Sea is an endangered species; this needs more investigation toward control or possible treatment of this helminth.

The present study was carried out to describe the epidemiologi­cal aspects of gastrointestinal helminthic infections of canids in Chaharmahal and Bakhtiari Province, the central western part of Iran. Forty nine canid species including, dogs, jackals, foxes and wolves were included in this study. The contents of their alimentary canal were in­spected in order to isolate and identify the parasitic helminthes of this system. To identify the worms, the Soulsbey and Anderson identification key and light microscopy were used. Based on necropsy findings, 35 (71.4%(of examined animals were infected with at least one helminth. The prevalence of identified worms was as follows: Mesocestoides lineatus (55.1%), Joyeuxiella echinorinchoides (26.5%), Tae­nia hydatigena (12.2%), T. multiceps (8.2%), T. ovis (2%), Dipylidium caninum (2%) and Spirura spp. (2%). No significant difference was noticed between the sam­pling areas, age and helminth infection. Only a significant difference was ob­served for prevalence of T. multiceps in wolf (25%), dog (21.4%), jackal and fox (0%) respectively (P < 0.05). The canids in Chaharmahal and Bakhtiari harbor several para­sites that some kind of them have zoonotic importance and may pose a threat to community health specially in rural areas.

Human linguatulosis poses an important medical and veterinary concern in en­demic countries. Animals, as reservoir host, play a major role in transmission of infestation and epidemiology of the disease. This study reports a case of human linguatulosis caused by Linguatula serrata in the city of Kerman, South-eastern Iran. A woman suffering from upper respiratory symptoms is presented. The pa­tient consumed raw liver of sheep who was admitted to the Afzalipour University Hospital in Kerman for the symptoms of upper respiratory tract. In microscopic examination of the nasopharyngeal discharge, L. serrata was detected. This report has future medical implication in precise diagnosis of L. serrata in patients with complaints of nasopharyngeal symptoms.

The objective of the present study was to survey the presence of Sarcocystis in sheep''s brain in North Khorasan Province. In general, 80 samples of sheep''s brain were collected from slaugh­tered sheep in slaughterhouses of North Khorasan Province. Tissue digestion method was used for observing bradyzoites in tissues. Histopathological pro­cessing tracing Sarcocystis and ensuing structural change in the brain tissue were conducted. PCR analysis was conducted on all the brain samples. Sequencing was done for one PCR product. Genotype was identified by Blast search and homology analysis. Sarcocystis spp. was found in one of the brain samples (1.25%) using tis­sue digestion method. The presence of bradyzoite was also confirmed in the pre­pared histopathological sections. PCR analysis was positive in one of samples. Genotyping of one sample proved that Sarcocystis species was Sarcocystis ovicanis and the nucleotide sequence of this parasite was deposited in the GenBank data­base under accession number No.KF489431. Sarcocystis ovicanis can involve brain tissue of sheep and conse­quently causes clinical symptoms.