mohammad abdigoudarzi

The saliva and salivary glands of ticks possess a wide range of immuno-pharmacologically active mole-cules that effectively modulate the activity of enzymes, antibodies, and amines that have a role in different biological processes. Derived components from saliva and salivary glands of hard ticks Ixodidae have been characterized as poten-tial natural sources for discovering promising anti-cancer drug candidates.

The anti-cancer activity of salivary gland extracts (SGEs) from Hyalomma anatolicum, Hyalomma drome-darii, Hyalomma marginatum, and Hyalomma schulzei was assessed. MTT assays and flow cytometry were done on the HT-29 colorectal cancer cell line to evaluate the anti-viability and proliferative inhibition.

Based on the MTT assay results, the SGEs from Hy. dromedarii had the highest and lowest substantial anti-viability effects on the HT-29 cancer cell and human foreskin fibroblast (HFF) normal cell, respectively. The cytometric assessment revealed a significant increase in the apoptosis and necrosis ratio of the HT-29 cancer cells after treatment with Hy. dromedarii SGEs.

The results demonstrated that Hy. dromedarii SGEs have significant anti-proliferative, anti-viability, and apoptotic potential. The result of this study suggests that Hy. dromedarii SGEs is an appropriate candidate for further investigations to identify and purify the mechanisms and molecules involved in the anti-cancer activity of the SGEs.

Cryptosporidiosis is a parasitic disease caused by the cryptosporidium protozoan in vertebrate animals. In livestock, especially ruminants, infants develop diarrhea syndromes. The infection is widespread throughout the world, as well as in Iran, which is reported from various species. Morphological diagnosis of Cryptosporidium species has many limitations and is of no taxonomic value alone, therefore using molecular methods helps remove these limitations to some extent. The current study is aimed at microscopic, molecular, and antigen detection and isolation of Cryptosporidium Parvum parasite. Firstly, there were 300 specimens collected from different parts of Iran. Next, purification of oocysts from feces was carried out according to the method of Casemore et al., through the flotation technique following which staining was done by means of the modified Ziehl-Neelsen method (Henriksen method), and identification through diagnostic keys. ELISA test was also performed on the specimens for which the results were from 1 positive to 4 positive. Results of our study demonstrated that from the 300 cases tested for Cryptosporidium through the molecular method, 48 cases (16%) were positive. These cases were collected during summer, autumn, and winter, however, more than 50% of positive cases were found among specimens collected in autumn. Furthermore, there were 54 positive cases found by means of PCR test, which indicates 6 cases more than that for ELISA results. Finally, results from PCR detection and ELISA, were subjected to Chi-Square analysis, where no significant difference between the collected data was observed (p=.0587).

Potomac Horse Fever (PHF) is an acute enterocolitis syndrome causing mild colic, fever and diarrhea in all life stages in horses. It is also responsible for abortion in pregnant mares. The agent of PHF was diagnosed as Neorickettsia risticii. PHF was first diagnosed as a livestock disease, by veterinarians in Maryland State of USA in horses living around Potomac River. Since, most of the cases of disease were recorded in July, August and September, then it was regarded as a seasonal disease. Later studies revealed that a trematode vector takes the role as a vector for transmission. Routine cell culture and PCR were used for clinical diagnosis of the PHF. Using indirect fluorescent antibody tests revealed positive cases in most places of Canada and America, which regarded as former contacts with the agent of disease. Neorickettsia risticii could not be seen in monocyte in prepared blood smears from clinical cases of horses and this is contrary to Anaplasma phagocytophilum which very easily seen in granulocytes of blood in infected horses. Neorickettsia risticii has been isolated from released trematodes from fresh water snails. PHF is not a contagious disease and infected horses could be kept near susceptible horses. PHF is not regarded as a zoonosis. It is concluded that related veterinarians to horse industry in Iran should be informed about the possible occurrence of disease.

Tropical Theileriosis caused by Theileria annulata is a tick-borne disease which transmitted by the ix odid tick members of the genus Hyalomma. Studies on different aspects of disease require to access infective sporozoite of parasite which produced by tick vector. This study was carried out to establish of T. annulata life cycle to achieve T. annulata infected ticks.

Laboratory rabbit and calf were used for rearing of Hyalomma anatolicum different instars. Unfed nymphs were fed on T. annulata infected calf. Clinical signs, Giemsa stained smears and Polymerase Chain Reaction (PCR) methods were used for detection of infection in blood and tick specimens. Susceptible calf was used for confirmation of sporozoites maturation and infectivity in bioassay test.

Hyalomma anatolicum two and three-host strategies of life cycle was lasted 90 and 116 days respectively. The PCR confirmed T. annulata infection in blood and tick samples. Maturation of T. annulata sporozoites was confirmed in bioassy test. First clinical symptom of disease was seen earlier in the case of transmission of disease through feeding of live ticks in comparison with blood injection method.

Complete life cycle of T. annulata was done and confirmed by clinical signs, microscopic examination, molecular methods and bioassay test. According to published reports to date, this is the first report of establishment of H. anatolicum tick infection with T. annulata using susceptible calf under controlled conditions in Iran.

The protozoan parasite Theileria annulata is the causative agent of tropical theileriosis in cattle. Vaccination is recommended by administration of attenuated schizont-infected cell lines. The expected protective immunity post-vaccination can be demonstrated by challenge test through inoculation of highly virulent infective sporozoites. The aim of this study was to produce Hyalomma anatolicum anatolicum tick infected with T. annulata (local strain) for preparation of tick-derived sporozoite stabilates for molecular characterization and infectivity test assay.

A local T. annulata strain was used for experimental infection of calves. A field isolate of H. a. anatolicum was isolated, laboratory-reared and infected by blood-feeding on Theileria infected above-mentioned calves. The infectivity of calf, tick and prepared stabilate were confirmed by clinical signs of theileriosis, microscopic inspection, RT-PCR and in vitro cell culture.

The tick stabilate was prepared and cryopreserved in liquid nitrogen. The infectivity of the tick stabilate was verified by in vivo bioassay, in vitro cell culture infection, microscopic inspection in salivary glands and RT-PCR assay. The in vitro produced cell line in this study was characterized by T. annulata Cytochrome b gene analyzing.

The infectivity of a new prepared tick-derived sporozoite stabilate was confirmed in susceptible calves; by microscopically, post mortem, tick microscopic and molecular assays. Moreover, naïve PBMCs were transformed and proliferated by T. annulata infected tick stabilate to immortal T. annulata schizont infected cell line. The potent infective sporozoite tick derived stabilate could be used for vaccine efficacy and challenge test as well as in vaccine development.

The identification of Hyalomma is a challenging issue in the systematics of ixodid ticks. Here, we examined 960 adult males of Hyalomma tick from 10 provinces of Iran using morphological and molecular methods. PCR was carried out on 60 samples to amplify an ITS2 fragment of nuclear and a COI fragment of mitochondrial genomes. Nine species, namely H. aegyptium, H. anatolicum, H. asiaticum, H. scupense, H. dromedarii, H. excavatum, H. marginatum, H. rufipes and H. schulzei were identified. The validity of H. rufipes and H. excavatum can be challenged. We concluded that these species should be regarded as H. marginatum and H. anatolicum complexes, respectively. Furthermore, the taxonomic status of the two closely related H. anatolicum and H. asiaticum was confirmed as distinct species.

Cette étude a été menée dans trois élevages bovins traditionnels de la région d’Ispahan (Iran) et avait pour objectifs l’évaluation de la diversité, dynamique saisonnière et abondance des tiques infestant les bovins. Au cours de cette étude, aucun acaricide n’a été administré et les tiques adultes ont été prélevées deux fois par mois à partir de différents endroits du corps des bovins. Les informations environnementales comme l’humidité et la température ambiante ainsi que la pluviométrie ont également été enregistrées lors de chaque prélèvement. Un total de 1206 tiques (56% de males et 44% de femelles) ont été prélevées des élevages sélectionnés. L’ Hyalomma anatolicumanatolicum représentait l’espèce la plus abondante et été suivie respectivement de l’ Hyalomma marginatummarginatum (8,7%) et de Rhipicephalus sanguinus (6,5%). L’abondance des tiques de la famille des Ixodidae montrait une nette augmentation, atteignant un pic important au printemps (Avril, Mai et Juin) en raison d’une activité accrue de l’H. anatolicumanatolicum. Cette espèce proliférait fortement en fin de saison printanière et au début de l’été. L’espèce R. sanguinusa été prélevée au mois de Mars (fin de l’hiver) ainsi qu’en juin (fin printemps). La plus forte concentration de tiques dures a été enregistrée au printemps alors qu’environ 27,1%, 3,5% et 1 % des tiques ont été respectivement collectées en été, automne et hiver. La majorité des tiques récoltées étaient localisées au niveau des mamelles (63,2%), suivies par la queue (36,1%) et les entrejambes (0,7%) alors qu’aucune tique n’a été détectée sur les autres parties du corps des bovins. Une corrélation significative a été observée entre l’abondance des tiques et la température ambiante moyenne(P

Ticks (Acari: Ixodidae) are obligatory blood feeders and important external parasites of wildlife, domestic animals and humans in tropical and subtropical areas. Ticks primarily transmit different pathogenic organisms to livestock and humans, and secondly harm their animal hosts by decreasing blood protein content and nutrients due to partial exsanguinations, induce allergic reaction and sometimes cause tick paralysis or toxicosis. In this study, ticks have been collected from different body parts of stable kept male and female camels (Camelus dromedarius) in monthly intervals from late April 2012 to late March 2013 in Masileh region of Qom province, Iran. All tick have been collected and identified according to male individual characters. Totally, 1265 ticks (42.5% and 57.4% from stable male and female camels respectively) have been collected. Tick identification based on male characters revealed Hyalomma dromedarii (40.7%), H. schulzei (40.3%) and H. marginatum (18.8%). Heavy camel infestation by H. schulzei was recorded for the first time from central part of Iran. Different tick infestation rates were recorded during June and January, from zero to 254 ticks. The highest mean number of collected ticks was 9.6 under tail of female camels among five body places during August. Statistical analysis showed a significant correlation between the number of collected ticks and sampling intervals, sex of camels and place of camel bodies. Out of five camel body places, ticks were collected regularly from four body parts. Periodical infestation by H. dromedarii under tail, belly, thigh and neck of both camel sexes was found to be longer than for H. schulzei and H. marginatum. It could be concluded that any control measure for camel tick infestation should be planned before March-April in Masileh region of Qom, Iran.

Biting midges of the genus Culicoides act as vectors for important diseases affecting humans and both wild and domestic animals. Collection of adult Culicoides specimens in the near vicinity of vertebrate hosts is the major part of any bluetongue surveillance plan. There are old records of Culicoides species dated from 1963, 1968 and 1975. Therefore, it was decided to collect different ceratopogonids members using a light trap.
One night catching using light traps with a suction fan was performed at representative sites (25 places) located in North Western Provinces (Ardebil, Eastern Azerbaijan, Western Azerbaijan and Zanjan) of Iran (suspected farms for clinical records of Bluetongue virus or serodiagnosis of the Bluetongue virus). Samples were detected and identified primarily and were sent to a reference center for final verification.
Seven Culicoides species including (Culicoides circumscriptus, C. flavidus, C. longipennis, C. pulicaris, C. puncatatus, C. nubeculosus, and three species from Culicoides (Oecacta) are under study in reference laboratory in Poland and C. puncticollis were confirmed from Iran.
Morphological and explanation of each species was regarded in this study. In comparison to old rec­ord, there are four new records of Culicoides species from Iran and one species is regarded suspected for viral trans­mission.

Hyalomma marginatum rufipes (Acari: Ixodidae) along with three closely related subspecies is considered as marginatum group. The subspecies had proven as main vector of Crimean Congo hemorrhagic fever virus, the cause of human death in Asia, Africa and Europe. This tick is also a vector of parasitic protozoan Theileria annulata, agent of tropical theileriosis in cattle. Nonetheless, taxonomical status of this tick not recognized or confirmed in tick’s fauna of Iran, then we decided to show most taxonomic characteristics and confirm the presence of this subspecies in Iran by molecular methods. Tick specimens were collected from cattle in Manujan township, Kerman province, southern Iran. Specimens were identified morphologically using suitable taxonomical identification keys. The morphologically identified specimens were subjected to molecular studies. Morphological and COI gene analysis clearly confirmed the occurrence of H. m. rufipes in Iran, however, according to ITS2 fragment H. m. rufipes can be the same H. m. marginatum. Thus, it seems that based on most popular molecular markers, H. m. rufipes and its relative H. m. marginatum really should be assigned as a polymorphic species H. marginatum.

This study was established to determine the lymnaea stagnalis snails’ infection with trematodes larval stage in one of the springs of the Shahrekord city in Chahar Mahal and Bakhtiari. To determine the snail infection to trematodes larval stages, the snails were caught from the field, and transferred to the Parasitology department of Razi Vaccine and Serum research Institute. Then stimulating of snails by light, tubing and squ ashing of them were used to detection and identification of the isolated cercariae. Of 400 collected Snails from the referred springs, 320 of them identified as lymnaea stagnalis. Observed cercariae were identified and classificated as order Plagi orchis, family plagiorchiidae and genus opisthioglyphe and plagiorchis. In Chahar Mahal and Bakhtiari province due to having more than 10% of water content of country, ecological conditions can play important role to developing sensitive snail especially L ymnaeidae and be considered as a critical and suitable habitat for them.

In order to study external parasites of laboratory reared animals at Razi institute, different methods including brushing of animal's surface body, cellophane tape of body surface, peri-anal cellophane tape test (CTT) and skin scrapings and digestive method were applied and collected samples were studied. In addition, field collected rats were tested using brushing method. One mouse had been infested by some mites. Rabbits, rats, mice and guinea pigs had not been infested with external parasites. Field collected rats had been highly infested with mites from the family Laelapidae. The, brushing method was confirmed to be a useful method for mite detection. According to the methods used in this study and these recommended by SOP from international animal breeding centers the CTT method was proposed to be useful for preparing health certificate of laboratory animals at the department of laboratory animal breading at Razi institute.

The poultry red mite, Dermanyssus gallinae is one of the most economically important ectoparasites in hens and some species of mammals worldwide. Cases of human infestation have been reported worldwide. In this study we report infestation in three members of a family referred with pruritus and allergic dermatitis rash. They have col­lected very small animals and carried them to the laboratory which later was confirmed as D. gallinae. They claimed that they had been bitten with this ectoparasite. This is the first case report of human infestation owing to D. gallinae from Iran.

Diagnostic study of vector ticks for different pathogens transmitted specifically have been done by Iranian old scientists working on the basis of biological transmission of pathogens. In this study we decided to con­firm natural infection of different collected ticks from three different provinces of Iran. Ticks were collected from livestock (sheep, goats and cattle) during favorable seasons (April to September 2007 and 2008). Slide preparations were stained by Giemsa and Feulgen and were studied searching for any trace of infection. Positive DNA from infected blood or tissue samples was provided and was used as positive control. First, PCR optimization for positive DNA was done, and then tick samples were subjected to specific PCR. Eleven pairs of primers were designed for detection of Theileria, Babesia and Anaplasma spp. Totally 21 tick samples were detected to be infected with protozoa. Hyalomma anatolicum anatolicum and Rhipicephalus turanicus from Fars Province were infected with T. lestoquardi at two different places. Hyalomma detritum was in­fected with T. lestoquardi in Lorestan Province and Rh. turanicus was infected to Ba. ovis from Fars Province. Totally 21 tick samples were detected to be infected with protozoa. Every sample is regarded with host-environment related factors. Since there are complex relations of vectors and their relevant protozoa, different proce­dures are presented for future studies.

The taxonomic status of ticks in the genus Hyalomma, as prominent vectors of domestic animal and human pathogen agents as well as hematophagous parasites of all terrestrial animals has a problematic history due to variability. The present paper is based on our observations on the Iranian Hyalomma species during 2009 to 2013. In this study, for nine Hyalomma species including H. aegyptium, H. anatolicum, H. asiaticum, H. detritum, H. dromedarii, H. excavatum, H. marginatum, H. rufipes and H. schulzei, morphologic characteristics and some notes on their variability, biology and distribution are presented. In this paper, diagnoses, host information, distribution data, illustrations of adult males and a taxonomic key to the native Hyalomma species of Iran are provided to facilitate their identification.

Theileriosis is caused by an intracellular protozoan that causes great economical losses to animal husbandry in different regions of Iran . The present study was done in order to identify of Theileriaovis in hard ticks in Lorestanprovince . Amongst five different regions in Lorestan, and during the April-to-july period of the year 2012, 265 cases of hard ticks were collected from the ear and the body surface and 100 blood smears from ear veins of anemic and feverish sheep. DNA extraction was done from the salivary glands of collected ticks and PCR test was performed using a pair of 520 bp specific primer of SSurRNA gene of T. ovis . The microscopic examinations of blood smears showed that 12 samples of blood smears ( 12 %) contained the piroplasmic forms of Theileriaspcies. The PCR revealed that 37 out of 152 Rhipicephalussanguineus (24/34 % ) were positive for T. ovisgenum including 21 female (13/81%) and 16 male (10/52%) ticks while the other ticks were not infected by this parasite. Out of total of 265 tick samples, R.sanguineus was highest (38.35%) and Haemaphysalispunctatahad the lowest (3/01%) frequencies. Regarding the vast distribution of R.sanguineus in the area, it seems that this tick may be the main vector of T.ovis in Lorestan province, Iran.

Tick paralysis is a disease of human and animals characterized by an acute ascending flaccid motor paralysis. The condition may terminate fatally unless the tick(s) are removed before respiratory paralysis occurs. Patient: A 48 years old female was referred to a clinic in Tehran with edema at the head and neck regions. She had fever and disequilibrium. She reported a travel to mountainous parts of Tehran in early spring. During physical examination a small tick was found attached firmly to the head. It has been detached and sent to Razi Research Institute and a female Dermacentor marginatus (Acari: Ixodidae) was verified. Following the tick removal the patient''s condition was improved. With respect to a rich fauna of Ixodid ticks in Iran, it is important to keep in mind the tick associated complications in this area.