Study of vitellogenin in Boophilus annulatus tick larvae and its immunological aspects.

Boophilus annulatus is an important one-host tick in the Mediterranean regions and Iran. It can transmit the Babesia bigemina, Babesia bovis and Anaplasma marginale to cattle. Nowadays, immunization programs by tick proteins is one of the potential methods for the control and prevention of tick infestations. Therefore, the characterization and identification of various tick proteins are necessary. Vitellogenin is a precursor of vitellin that is produced in mid gut cells and fat bodies in ticks. In this study, we characterized vitellogenin protein of B. annulatus unfed larvae using one- and two-dimensional electrophoresis and immunoblotting. In one-dimensional immunoblotting, 48, 70, 100, 130 and >250 kDa protein bands positively reacted with immune sera. In two-dimensional immunoblotting many protein spots positively reacted with immune sera. Six of them were analyzed by MALDI-TOF and MALDI-TOF- TOF mass spectrometry. The results showed that amino acid sequences of four immunogenic proteins with molecular weights of 38, 43, 85 and 97 kDa had identity to tick vitellogenin and its homologues (GP80), based on the Mascot search results. It seems that more knowledge on tick proteins including vitellogenin and their characterization could be useful for the development of anti-tick vaccines.

Evaluating the resistance of eimeria spp. Field isolates to anticoccidial drugs using three different indices.

BACKGROUND: In this study, the presence of resistance to diclazuril, amprolium+ethopabate and salinomycin, representing some of the commonest anticoccidials in Iran's poultry industry, against three mixed Eimeria field isolates were investigated. METHODS: Three Eimeria field isolates, collected from typical broiler farms in Iran, were propagated once, inoculated to 480 broilers, comprising 30 chicks in each treatment. The non-medicated or medicated diets containing one of the above mentioned anticoccidials were provided ad-lib. Drug efficacy was determined using the Global index (GI), Anticoccidial Sensitivity Test (AST) and Optimum Anticoccidial Activity (OAA). RESULTS: None of the field isolates were fully sensitive to the selected anticoccidials. All isolates showed reduced sensitivity/partial resistance to salinomycin. Resistance to amprolium+ethopabate was evident and partial to complete resistance was recorded for diclazuril. CONCLUSION: Limited efficacy of the selected anticoccidials is obvious. Considering the cost of continuous use of anticoccidials in the field, altering the prevention strategy and rotation of the anticoccidials with better efficacy, would prevent further economic losses induced by coccidiosis.

Identification of Tropomyosin and Its Immunological Properties from Larvae of Cattle Tick, Boophilus annulatus.

BACKGROUND: Boophilus annulatus is an obligate blood feeder tick that can cause great losses in animals due to anemia and its ability to injure its host skin directly. The aim of this study was identification of cattle humoral immune response to some tick proteins during experimental infestation. METHODS: Immune sera against tick were collected from experimentally infested cattle with ticks. One and two-dimensional electrophoresis and Western blotting methods were used for the detection of immunogenic proteins in larval tick extract and eight of these proteins were identified by MALDITOF and MALDI-TOF-TOF mass spectrometry. RESULTS: In non-reducing one-dimensional SDS-PAGE, some bounds between 12 to more than 250-kDa appeared. In two-dimensional SDS-PAGE, numerous spot appeared and the identified immunogenic proteins by parallel immunoblotting weighted between 14 and 97 kDa. Amino acid sequences of protein spot with 37-kDa molecular weight had identity to tropomyosin based on Mascot search in NCBI. CONCLUSION: Anti tropomyosin antibodies can be induced in experimentally infested hosts with ticks and it seems that tropomyosin can be useful for the development of anti tick vaccines.

Analysis of Immunogenic Relevant Proteins in Rhipicephalus (Boophilus) annulatus Tick.

BACKGROUND: Considering the importance of ticks as a main group transmitting pathogen organisms, this study designed to recognize immunogenic proteins in different tissues of Rhipicephalus (Boophilus) annulatus tick and to find out if there are common proteins in these tissues. METHODS: Seven cattle were experimentally infested with about 10000 R. annulatus larvae and their humoral immune response to extracts of salivary gland and ovary of adult ticks and larval extracts during infestation were determined by ELISA and Western blot analysis. Measurements of serum antibodies level recorded weekly, from week 0 to week 9. RESULTS: Using Western blot analysis, 15 fractions from soluble antigens extracted from salivary gland and larvae, and 14 fractions in the larval extracts were recognized. These findings illustrate the recognition of common antigens with molecular weight of 170, 117, 100, 70, 37, 33 and 30 kDa from different antigens by resistant cattle sera. CONCLUSION: Common antigens are present in different tissues of Rhipicephalus (Boophilus) annulatus, which can be used as a target in immunization against ticks.

First Detection of Nosema ceranae, a Microsporidian Protozoa of European Honeybees (Apis mellifera) In Iran.

BACKGROUND: Nosemosis of European honey bee (Apis mellifera) is present in bee colonies worldwide. Until recently, Nosema apis had been regarded as the causative agent of the disease, that causes heavy economic losses in apicultures. Nosema ceranae is an emerging microsporidian parasite of European honeybees, A. mellifera, but its distribution is not well known. Previously, nosemosis in honeybees in Iran was attributed exclusively to N. apis. METHODS: Six Nosema positive samples (determined from light microscopy of spores) of adult worker bees from one province of Iran (Savadkouh- Mazandaran, northern Iran) were tested to determine Nosema species using previously- developed PCR primers of the 16 S rRNA gene. As it is difficult to distinguish N. ceranae and N. apis morphologically, a PCR assay based on 16 S ribosomal RNA has been used to differentiate N. apis and N. ceranae. RESULTS: Only N. ceranae was found in all samples, indicating that this species present in Iran apiaries. CONCLUSION: This is the first report of N. ceranae in colonies of A. mellifera in Iran. It seems that intensive surveys are needed to determine the distribution and prevalence of N. ceranae in different regions of Iran.

Biopathologic Characterization of Three Mixed Poultry Eimeria spp. Isolates.

BACKGROUND: Coccidiosis of domestic fowl, caused by species of the Genus Eimeria, is responsible for important economic losses in poultry production. Because different species and/or strains can vary in pathogenicity and other biological parameters, their precise characterization is important for epizootiological studies. METHODS: Fifty samples from litter, whole intestinal tract and feces were collected from poultry houses located in different provinces of Iran. One hundred twenty male day-old broiler chicks were challenged with three selected isolates. Data on weight gain, Food Conversion Ratio (FCR), food intake, lesion scoring and shedding of oocysts per gram of feces were recorded and analyzed by the Duncan's test. RESULTS: In all treatments, the challenged groups had statistically significant lower weight gain than that of unchallenged control group. Isolate three caused the lowest weight gain and food intake and the worst lesion score as well as FCR. Despite originating from close geographical regions for isolates 1 and 2, the difference in biopathologic factors may be either due to different proportion of identified species or the different pathogenicity of the species present in the isolates. CONCLUSION: The results highlight the importance of considering various species of Eimeria in designing the preventive, control and treatment strategies to prevent coccidiosis in different regions of Iran. Further characterization of each isolate would be the next step to provide a basis for coccidiosis research with well-characterized local isolates.

Molecular and biomorphometrical identification of ovine babesiosis in iran.

BACKGROUND: Ovine babesiosis is the most important haemoparasitic tick-borne disease of small ruminants in Iran caused by Babesia ovis, B. motasi, and B. crassa. The aim of this study was to characterize the species of ovine Babesia species isolated from different geographical region of Iran. METHODS: One hundred fifty four blood samples collected from animals, which demonstrated the pale mucous membranes or hyperthermia. The specimens were transferred to the laboratory and the blood smears stained with Geimsa, the morphological and biometrical data of parasite in any infected erythrocyte have been considered. Extracted DNA from each blood samples were used in PCR and semi nested- PCR in order to confirm the presence of the species. RESULTS: Microscopical observation on 154 blood smears determined 38 (24.67%) and 40 (26%) samples were infected by Babesia and Theileria respectively. The mixed infections occurred in four (2.6%) samples. The results of the PCR assays showed nine (5.85%), 81 (53%) and 18 (11.7%) were distinguished as Babesia, Theileria and mixed infection, respectively. Semi nested- PCR did not confirm the presence of B. motasi. CONCLUSION: The causative organism of many cases of haemoprotozoal diseases, which recorded in previous studies, could be B. ovis or Theileria lestoquardi. The result confirmed that B. ovis was only species which causes babesiosis in the study areas. It seems that the biometrical polymorphisms could exist in B. ovis in Iran. This polymorphism could be a main problem in differentiation between B. ovis and B. motasi and it could be dissolved by specific PCR analysis.

One-Humped Camel (Camelus dromedarius) Infestation with Linguatula serrata in Tabriz, Iran.

BACKGROUND: Linguatula serrata is one of well known members of Pentastomida which infects both human and animals. The aim of this study was to evaluate the prevalence of L. serrata in mesenteric lymph nodes, livers and lungs of camels slaughtered in Tabriz area, Iran. METHODS: Mesenteric lymph nodes (MLNs), livers and lungs of 140 one-humped camels slaughtered in Tabriz, north-west of Iran were investigated for nymphs of L. serrata from July 2007 to June 2008. The organs were examined macroscopically and then a tissue digestion method was also done for investigation of liver and lung of the camels that had infected MLN. The liver and lung samples were mostly taken from condemned and rejected part of organs. RESULTS: The infection rate of L. serrata nymphs in MLNs, livers and lungs was 13.5%, 1.4% and 1.4% respectively. The number of isolated nymph in infected lymph nodes varied from 2 to 18 with a mean of 4.78. Only one nymph was isolated from each infected livers and lungs. The infection rate increased with age (P< 0.05). No significant difference in different sex groups and seasons was observed (P> 0.05). CONCLUSION: Considering this fact that consumption of undercooked camel liver was not common in the studied area, the zoonotic importance of this infection should be concluded.

The distribution of Hyalomma spp. ticks from domestic ruminants in Iran.

The distribution of Hyalomma species on domestic animals was studied in four zoogeoghraphical zones. Nine hundred and ninety-two Hyalomma ticks were collected from sheep, goats, cattle and camels. A total of seven tick species consisting of: Hyalomma anatolicum Koch, 1844 (28.93%), H. excavatum Koch, 1844 (12.5%), H. asiaticum Schulze & Schlottke, 1930 (13.5%), H. marginatum Koch, 1844 (27.01%), H. detritum Schulze, 1919 (9.67%), H. schulzei Olenev, 1931 (4.03%) and H. dromedary Koch, 1844 (4.33%) were recorded. The results indicated that H. anatolicum, H. asiaticum, H. marginatum and H. detritum were present in all zones whereas H. excavatum was absent in Zone I. The result also showed that H. dromedari was presented in Zones III and IV whereas H. schulzei was present in Zones II and III. During this study, the tick species collected are candidates for investigation as vectors of pathogenic organisms in Iran.

Some observations on sheep sarcoptic mange in Tehran province, Iran.

Sarcoptes scabiei infestation was diagnosed in 278 sheep from 12 fatling flocks. The sheep presented crusted skin lesions initially appear on the lips or nostrils, the lesion on nostrils also extend towards around the eyes, the supraorbital fossae and in some cases over entire face. The infested male lambs by mating behavior in fatling flocks caused transmitting the infection to fat tail area and scrotum. Histopathological study of lesions demonstrated marked acanthosis, hyperkeratosis, parakeratosis. Tunnels could be observed in the hyperkeratotic stratum corneum and mite segments were located mainly in the stratum corneum and also in the stratum granulosum. Attempts to eliminate S. scabiei var. ovis were made in 3 naturally infested sheep herds, by two dippings with two weeks interval with Amitraz, Cypermetrin and Prompetamphos. The results of this study indicate that acaricidal treatment of S. scabiei var. ovis in 3 naturally infested herds was successful, but a few cases of reinfestation were found in each treated group.

Haematophagus mites in poultry farms of iran.

BACKGROUND: Blood sucking mites are important avian ectoparasites which being found on bird species worldwide. Their presence are problematic for the producers either through potential direct effects on weight gain, egg production, sperm production in roosters or as nuisance pests on worker handle hens and eggs. The aim of this study was pointing out of the status of haematophagus mites. METHODS: Eight caged layer and four breeder flocks were visited, monitoring for the presence of chicken mites performed by removing and examining debris from poultry house, infested nesting material collected into zip lock plastic bags and at least 20 birds were also randomly selected to examine the presence of chicken mites. Mites obtained from each population were mounted in Hoyer's medium on microscope slides and identified. All eight caged layer and four breeder flocks were inspected, which were infested with chicken blood feeding mites. RESULTS: Massive infestations of Dermanyssus gallinae were common with huge numbers of parasites on birds, cages and the conveyor belts for egg. Only one farm from Mazandaran Province was infested to Ornithonyssus bursa. CONCLUSION: Dermanyssus gallinae was the most prevalent blood feeder mite in the breeder and caged layer flocks in Iran, while O. bursa was reported as a first record, which found only in a breeder flock in Mazanderan Province. It seems that its presence is limited into the area which affected by both warm and humid environmental conditions.