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Introduction: The enteric protozoa, Giardia duodenalis (G. duodenalis), consists of eight distinct assemblages (A-H) with identical morphological characteristics and a direct life cycle. Successful axenic cultivation of this parasite is an important preliminary step for biological, drug resistance and phylogenetic studies. Moreover, G. duodenalis exhibits great genetic and biotypic diversity. The current study aimed to evaluate In vitro culture and multilocus genotyping of G. duodenalis trophozoites obtained from human fecal samples in southwest Iran. Methods: Thirty human fecal specimens containing G. duodenalis cysts were collected from Ahvaz city (southwest of Iran). The purification of cysts was carried out by the sucrose flotation technique. The cysts were inoculated in a modified TYI-S-33 medium and was daily monitored for the development and viability of trophozoites. After extracting DNA, gdh, bg and tpi genes were evaluated using molecular techniques (the semi-nested PCR for gdh gene and the nested PCR for tpi and bg genes). Eventually, the amplified fragments were sequenced and then, the phylogenetic tree was drawn. Results: Of 30, the trophozoites were encysted from five samples. All three genes were detected in two cases of five samples using molecular techniques. The multilocus phylogenetic analysis demonstrated that all the two samples belonged to assemblage A and sub-assemblage AІІ. Conclusion: Our findings indicated the presence of different numbers of trophozoites with variable development and survival rates in modified TYI-S-33 medium. Furthermore, the multilocus genotyping showed that these trophozoites belonged to assemblage A and sub-assemblage AІІ.

Objective Contamination of soil and water with infected oocysts have an important role in both animals and humans toxoplasmosis. Cats shed oocytes by their feces that can survive in environment such as moist soil, warm and humid weather for months and even years. The present study designated to molecular detection of Toxoplasma gondii oocysts in stool samples obtained from cats in Ahvaz, southwest of Iran. Methods Initially, 486 stool samples were randomly collected from cats. After sucrose flotation method, the DNA was extracted and PCR was carried out using the amplification of the B1 gene and repeat element sequence (RE). For strain typing purpose, the genetic marker of SAG2 was used in nested-PCR. To perform RFLP procedure, the products of nested-PCR were digested using Sau3aI and HhaI enzymes. Results A total 486 stool samples were examined to the amplification of the 194 bp fragment of B1 gene that 35 samples (7.2%) were positive. All positive samples were confirmed by using another PCR that was amplified the 130 bp repeat element sequence (RE). The identified genotypes were type III (32 cases), mix of type I and III (2 cases) and one of the samples was type I. Conclusion Our findings revealed a relatively high prevalence of T. gondii oocysts in stool samples obtained from cats. It is essential that the high-risk people receive the information about the risk of direct and indirect contact with the animals.

Rove beetles (Coleoptera: Staphilinidae) are a group of diverse insects occurring in a range of different ecosystems, from the plains to the mountaintops. However, there are no reports in the literature of their presence in the tidal zone of the seashore. This is the first report of the presence ofPaederus fuscipesCurtis on the coastal sands of the Caspian Sea, northern Iran. In all 14 counties of Mazandaran Province on the southern coast of the Caspian Sea, systematic monthly sampling of the beetles took place from May 2012 to April 2013. Samplings (30-min-person) were performed and specimens collected from the sandy seashore by gloved bare hands and identified using appropriate keys. To our surprise, all 1012 specimens collected were exclusivelyP. fuscipesCurtis, but with significant seasonal and geographical variations among the numbers of specimens collected. The distribution and activity ofP. fuscipeswere outstanding in Mahmoodabad and Fereidonkenar counties, with the highest abundance and frequency in September, posing a serious health threat to tourists and residents of these areas.

Resistance to acaricides in ticks is becoming increasingly widespread throughout the world; therefore, tick control requires resistance monitoring for each tick species. The aims of this study were to monitor the susceptibility status of the cattle tick, Rhipicephalus (Boophilus) annulatus (Acari: Ixodidae), against pyrethroid acaricides from Mazandaran Province, northern Iran, and where resistance was evident, and establish the possible underlying mechanisms. Fully engorged adult R. (B.) annulatus females collected on cattle from Mazandaran Province. Twenty-nine tick populations produced 10-18days old larvae and bioassayed with cypermethrin and λ-cyhalothrin by larval packet test and the levels of detoxification enzymes were measured. Population AM-29 had a maximum resistance ratio (RR99) of 20.21 to cypermethrin and 53.57% of the tick populations were resistant at LC99 level. With λ-cyhalothrin, 17.86% of the tick populations were resistant and AM-29 was the most resistant population with RR99=4.54. AM-29 also showed significant elevation of cytochrome P450 monooxygenase (P450) and glutathione S-transferase (GST) (2.76- and 2.39-fold, respectively) (P

Rhipicephalus (Boophilus) annulatus is one of the most important hard ticks parasitizing cattle in northern Iran. The aim of this study was to evaluate pyrethroid resistance levels of this species from Nur County, northern Iran. The hard ticks were collected through a multistage cluster randomized sampling method from the study area and fully engorged female R. (B.) annulatus were reared in a controlled insectary until they produced larvae for bioassay. Seventeen populations of the hard ticks were bioassayed with cypermethrin and 12 populations with lambda-cyhalothrin using a modified larval packet test (LPT). Biochemical assays to measure the contents/activity of different enzyme groups including mixed function oxidases (MFOs), glutathione S-transferases (GSTs) and general esterases were performed. Population 75 showed a resistance ratio of 4.05 with cypermethrin when compared with the most susceptible field population 66 at the LC50 level. With lambda-cyhalothrin the resistance ratio based on LC50 was 3.67 when compared with the susceptible population. The results of biochemical assays demonstrated significantly elevated levels of GSTs and esterases in populations tested compared with the heterozygous susceptible filed population and a correlation coefficient of these enzymes was found in association to lambda-cyhalothrin resistance. Based on the results, pyrethroid acaricides may operationally fail to control R. (B.) annulatus in North of Iran. This study is the first document of pyrethroid resistance in R. (B.) annulatus populations from Iran.

Background: Nowadays a number of methods are used for the extraction of genomic DNA from fecal specimens. Identification and selection of an effective DNA extraction method is considered as one of the most important steps in molecular assays of Giardia duodenalis. Objectives: We compared the effects of 3 different DNA extraction techniques in PCR amplification of a specific area of SSU rRNA gene. Methods: A total of 20 fecal samples containing purified cysts were aliquoted in 3 sub-samples. DNA extraction was performed using the Phenol-Chloroform Isoamyl alcohol (PCI), QIAamp DNA stool mini kit, and YTA Stool DNA Isolation mini Kit. The quantity and purity of extracted DNA were compared. The potency of extracted DNA was tested. Results: The results showed that the most concentrated DNA was obtained from phenol/chloroform/Isoamyl alcohol method and the best purity based on comparing the ratio of A260/230 was obtained from QIAamp DNA stool mini kit. In this study the diagnostic sensitivity of QIAamp DNA stool mini kit, phenol/chloroform/Isoamyl alcohol, and YTA Stool DNA Isolation mini Kit methods was 60%, 70%, and 60%, respectively. Conclusions: The application of a proper DNA extraction method leads to obtaining reliable and reproducible results in molecular assays and supports treatment and control strategies of G. duodenalis. In this study, PCR amplification targeting a 350-bp fragment of SSUrRNA gene demonstrated phenol/chloroform/Isoamyl alcohol as the most efficient method.

Background: Cryptosporidium is a protozoan parasite that effects rodents, dogs, calves, humans, and cats. Infection with this parasite is known as cryptosporidiosis. Cryptosporidium spp. may induce clinical or subclinical signs in infected hosts. In the life cycle of this parasite infected dogs freely living in urban and rural areas of Khuzestan province are the definitive hosts that should be considered as a real problem in public health for humans. Objectives: This study aimed at determining the frequency of cryptosporidiosis in dogs in southwest of Iran. Methods: Overall, 350 fresh fecal samples were collected from domestic dogs living in 43 villages, from June 2012 to September 2013. All samples were investigated by Sheather’s concentration method and fecal smears were stained with modified Ziehl-Neelsen followed by light microscope examination, and polymerase chain reaction (PCR). Results: The results revealed that frequency of Cryptosporidium infection was 8% and 12.3%, using direct smear and molecular method, respectively. Conclusions: The present findings indicated that domestic dog feces from southwest of Iranmay contain zoonotic parasites such as Cryptosporidium spp. andmay be a potential risk for humans and other animals, especially when they contaminate the environment. The role of dogs as source of human infection should be investigated by further studies.

Parasitic infection is highly prevalent throughout the developing countries of the world. These infections are the major problem in rehabilitation centers for the mentally retarded. There have been many reports about the prevalence of parasitic infection among different groups of people in Iran; however, the epidemiological data in intellectual disability (ID) individuals are rare and there are no such data from northern Iran. The aim of the present study was to determine the incidence of intestinal parasitic infections among the inhabitants of rehabilitation centers of Mazandaran province, northern Iran. A triple fecal specimens was collected from each of the 362 participants (183 males, 179 females) residing in six rehabilitation centers of Behshahr, Sari, Amol, Nowshahr and Ramsar. Each specimen was examined by direct wet mounting, formol-ether concentration, Ziehl-Neelsen and trichrome permanent staining. The overall infection rate of intestinal parasite was 26.2% (95 out of 362 subjects; 22.4% in males and 30.2% in females). Giardia lamblia was the most commonly found protozoan parasite (8.0%) followed by Entamoeba coli (5.5%), Blastocystis hominis (3.3%), Endolimax nana (2.8%) and Entamoeba histolytica (1.7%). Double infections were E. coli and B. hominis (1.1%), G. lamblia and Chilomastix mesnili (1.1%), G. lamblia and B. hominis (0.5%), G. lamblia and E. coli (0.5%), E. histolytica and E. coli (0.5%), E. nana and E. coli (0.5%) and E. nana and B. hominis (0.5%). No egg-positive case was found. These helminthic parasites were found to be not common among the participants, 75% of whom had been treated with mebendazole by guardians as a tradition, every 6 months. Nevertheless, our data in rehabilitation centers in northern Iran suggest that infection is not higher than that in community population in recent years in this geographical area.

Rhipicephalus bursa is one of the most common hard tick species in Iran. It is a vector of animal and human diseases. Tick control strategies in Iran rely heavily on pyrethroid insecticide use. Hence, susceptibility status of multiple field collected populations to λ-cyhalothrin and cypermethrin was investigated using the FAO recommended larval packet test. Resistance ratios at the LC 99 of field populations of R. bursa compared to a susceptible strain ranged from 0.95 to 4.78 for λ-cyhalothrin and 1.43–8.6 for cypermethrin. They were 2- to 6.9-fold higher than the maximum dose recommended by the acaricide formulating companies. Biochemical assays on different field collected populations showed significant elevation in esterase and glutathione S -transferase activities and monooxygenase contents compared with the susceptible strain. Therefore, close monitoring and resistance management strategies should be employed to delay the operational loss of pyrethroids for tick control in Iran.

Hard tick infestation is an important problem in both traditional and industrial animal husbandry as well as in veterinary and public health. Several acaricides have been used to control tick infestation in Iran. Poor control has been attributed to acaricide resistance in tick populations although this has not been demonstrated experimentally. In this study, susceptibility status to propetamphos (Blotic) of 2-3-week-old larvae of Rhipicephalus bursa, one of the most common tick species in Iran, was evaluated using the FAO recommended larval packet test (LPT) method with some modifications. Log dosage probit analysis of propetamphos dose-response regression lines produced a resistance ratio based on LC(50) in the most resistant strain of approximately 103-fold. Biochemical assays suggested that the tick strains contained multiple insecticide resistance mechanisms involving elevation of esterases, GSTs, MFOs and alteration of acetylcholinesterase. In light of this data, practical implications for pest and pesticide resistance management strategies are discussed.