Veterinary Research Forum

Abstract Nosema disease is one of the most important diseases of adult honey bees worldwide. It is known as silent killer because there are no characteristic symptoms. The aim of the present study was to determine prevalence of Nosema species in various towns of Kurdistan province in Iran. A multiplex polymerase chain reaction (multiplex-PCR) was performed for identification of Nosema species infecting European honeybee, Apis mellifera. A total of 100 samples were collected from apiaries (870 hives) in 10 counties of Kurdistan province, located in the west of Iran. Samples were examined using light microscope and PCR. The light microscope was used to determine the presence of Nosema spores in all of the collected samples. Multiplex-PCR based on 16S ribosomal RNA was used to differentiate N. apis from N. ceranae. Overall prevalence of the microscopic evaluation and PCR method were 29.00% and 32.00%, respectively. The analysis of Nosema isolates from interrogation of DNA databank entries of Kurdistan apiaries (based on rRNA sequence data) indicated that only N. ceranae was widespread in these apiaries, and it had already been found in high percentages (50.00%) in Marivan and Kamiaran counties of Kurdistan province. It was shown that only N. ceranae was found by PCR assay in the region.

Abstract Monogenea is one of the most species-rich groups of parasitic flatworms worldwide with many species described from African freshwater fish. Little is known about the diversity and geographic distribution of monogenean parasites infesting the Red Sea fishes in Egypt. In the present study, a total of 45 specimens of the brushtooth lizardfish Saurida undosquamis (family: Synodontidae) and 35 specimens of the red porgy seabream Pagrus pagrus (family: Sparidae) was examined for monogenean infestation. Samples were collected from water locations at Hurghada coasts along the Red Sea in Egypt. Two different species were recovered. The first recorded parasite was Diclidophora merlangi infesting the lizardfish. This parasite was morphologically similar to the original description for the general body shape, size, shape and arrangement of the clamps and reproduction organs and the number of spines in the lateral groups of the genital atrium, but is distinguished in the host fish which is of a different genus. The second species was Loxuroides pricei. The morphological and quantitative data of the isolated specimens and the potential reproductive consequences supported their assignment to L. pricei than to the other congeneric species.This parasite can be separated from the morphologically similar L. sasikala through having a shorter distance from the anterior extremity to genital atrium or vaginal region, fewer testes and a slightly greater number of spines on cirrus and genital atrium. The two species represented new host and locality records from the Red Sea in Egypt.

Abstract Giardia duodenalis is one of the most prevalent intestinal protozoa infecting humans and domestic animals. The aim of this study was to identify subspecies of G. duodenalis by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method from fecal samples of naturally infected cattle in the Urmia, West Azerbaijan province, Iran. Overall, 246 fecal specimens were collected from the cattle (diarrheic and healthy) and microscopically examined for G. duodenalis. The PCR-RFLP analysis of glutamate dehydrogenase (gdh) locus was used to identify the genotypes found in cattle. In this method, 432 bp expected size was amplified and then specific restriction NlaIV enzyme was used for subspecies detection. Totally, 23 (9.34%) specimens were microscopically positive for giardiacyst out of 246 examined samples. The PCR-RFLP analysis revealed that 19 samples (82.60%) have the genotype E and 4 samples (17.39%) belong to the subgroup AI. Our findings indicated that G. duodenalis infection is prevalent in cattle of Urmia and the non-zoonotic genotype E predominates in cattle in this region.

Abstract A total number of 450 blood samples were collected from 45 different randomly selected cattle herds. Light microscopic examination of blood smears revealed Babesia spp. infection in 4.2%, while 8.9% of blood samples were positive using PCR. Upon multiplex-PCR (mPCR), B. bigemina and B. bovis infections were detected in 37/40 (92.5%) and 3/40 (7.5%) samples, respectively. 530 ticks of 10 Ixodid species were collected from the same cattle. Hyalomma anatolicum was the most prevalent tick species (19.9%). An expected 520 bp fragment of Babesia spp. was generated in 22 (48.8%) of Rhpicephalus annulatus, 18 (40.0%) of R. bursa and 12 (30.0%) R. sanguineus sensu lato. The mPCR findings revealed that all infected ticks including R. annulatus, R. bursa and R. sanguineus were totally infected with B. bigemina. The DNA amplification of B. bovis and B. bigemina in egg samples showed that only B. bigemina was detected in two specimens of R. annulatus. It could be concluded that B. bigemina was the dominant causative agent in this region but the evidence of B. bovis infection of cattle in a few cases was noted, as well. The results suggested that B. bigemina and B. bovis could be detected in the DNA extracted from R. annulatus, R. bursa and R. sanguineus sensu lato confirming previous reports. Since B. bigemina is transmitted transovarially by R. annulatus, it might act as an important vector for B. bigemina.

Abstract Ehrlichiosis is a zoonotic disease which has been reported from some regions of Iran. This study was aimed to determine the presence and prevalence of ehrlichiosis in suspected dogs referred to the Faculty of Veterinary Medicine, Shiraz University, Shiraz, Iran using polymerase chain reaction(PCR). Blood samples were collected from 98 suspected dogs with at least one of the five following findings: thrombocytopenia, anemia (hematocrit < 37.00%), gastrointestinal signs and respiratory and/or central nervous system diseases. Complete blood count was performed for each sample. After genomic DNA extraction, PCR assay was carried out using a commercial PCR kit. The results showed that only three out of 98 samples (3.06%) were positive for ehrlichiosis. There was no significant difference in hematological parameters between infected and non-infected cases. These results emphasize that ehrlichiosis has a low prevalence among examined cases in southern Iran. Further serological and molecular studies are needed to clarify the epidemiological feature of this infection in different areas of Iran.

Abstract There is no prospective study from Iran to estimate the direct risk of Neospora caninum for pregnancy loss or reproductive factors. In addition, there is no report in the literature concerning the association of N. caninum with dystocia and sex of calves. Therefore, this study was conducted on a group of dairy cows in a large intensive production system during 2011 to 2013 in southern Iran to evaluate the impact of neosporosis on reproductive performance. A total of 253 cows which were diagnosed as pregnant during the first six months of the study were followed until calving or abortion. Reproductive data were collected and N. caninum serostatus was determined using ELISA. To investigate the association of abortion with N. caninum,survival analysis was performed using Cox proportional hazard model. The N. caninum seroprevalence in the study group was 30.40% (95% CI: 27.40, 36.10). The overall abortion rate was 12.25%, significantly higher in seropositive animals (20.80%) than seronegative ones (8.50%). Results of Cox model showed that serostatus of animal for N. caninum and season had significant associations with abortion (p < 0.01). Neospora caninum did not show significant association with other factors such as dystocia and sex of calves. In conclusion, neosporosis is responsible for 12.00% excess abortion risk in infected group and more than 30.00% of abortions could be preventable by control of Neospora in study population. Therefore, control of N. caninum would reduce the economic losses caused by parasite mainly due to pregnancy loss.

Abstract This study was aimed to determine the infection rate and vectors of Theileria lestoquardi in goats from West Azerbaijan province, Iran. A total of 400 blood samples were collected from 40 randomly selected flocks in the study area from June to September, 2014. Out of 400 blood samples examined using microscopic examination, a number of 14 goats (3.50%) were positive for Theileria spp., whereas 25 goats (6.25%) yielded a specific T. lestoquardi SSU-rRNA fragment (235 bp). The prevalence of theileriosis in goats estimated by semi-nested PCR was significantly higher than the prevalence estimated by microscopic examination of the blood smears. The prevalence of Theileria infection in different age and sex groups of goats was not significantly different. The highest and lowest prevalence of Theileria infection was in July (12.00%) and September (2.00%), respectively. A number of 315 adult Ixodid ticks were also collected from naturally infested goats and they were characterized. Out of 315 examined ticks, a number of 37 ticks including Hyalomma marginatum (65.20%), Rhipicephalus turanicus (44.00%), and Dermacentor marginatus (68.70%) were infected by T. lestoquardi. Based on the obtained results, it was concluded that the semi-nested PCR assay based on SSU-rRNA gene is a valuable method for epidemiological investigation of caprine theileriosis. The results showed that H. marginatum, R. turanicus and D. marginatus can be considered as risk factor in the epidemiology of T. lestoquardi.

Abstract Dirofilaria immitis is an important filarial nematode in dogs. In this study, age and sex distribution of this zoonotic nematode among dogs were investigated in northwest of Iran in Meshkin-Shahr city. Molecular characteristics of the isolates, based on cytochrome oxidase subunit 1 (COX1) gene were compared to the isolates from other areas of the world.Blood samples were collected from 91 dogs which were selected by simple classified accidental sampling. Thin and thick blood smear examinations were used to find out infectivity with D. immitis. DNA extraction was performed from adult D. immitis recovered from heart of infected dogs. The COX1 gene was amplified and sequenced. Phylogenetic analysis was carried out using sequences obtained in this study along with relevant sequences deposited in the GenBank. Phylogenetic analysis and sequence variation was performed using MEGA software in comparison with those COX1 sequences deposited in GenBank. Out of 91 dogs, 19 (20.87%) were found positive for infection with D. immitis. There was no statistically significant difference between males and females of dogs in terms of D. immitis infection. However, the rate of infection in dogs more than 2 years old was significantly higher than those with lower age. Both sequences analyzed in this study showed 100% homology to each other. Intra-species variation of these isolates with those from other areas of the world amounted to 0 to 0.50%. Phylogenetic analysis of the COX1 gene suggested that it is conserved, and can be used for study on genetic diversity and classification of filarial nematodes.

Abstract Toxocara canis (Nematoda: Ascaridae) is an intestinal nematode parasite of dogs, which can also cause disease in humans. Transmission to humans usually occurs because of direct contact with T. canis eggs present in soil contaminated with the feces of infected dogs. This nematode has extraordinary abilities to survive for many years in different tissues of vertebrates, and develop to maturity in the intestinal tract of its definitive host. Survival of parasitic nematodes within a host requires immune evasion using complicated pathways. Morphine-like substance, as well as opioids, which are known as down regulating agents, can modulate both innate and acquired immune responses, and let the parasite survives in their hosts. In the present study, we aimed to find evidences of morphine-like substance and µ-opiate receptor expression in T. canis, using high performance liquid chromatography (HPLC) and reverse transcription polymerase chain reaction (RT-PCR). The results indicated that T. canis produced morphine-like substances at the level of 2.31± 0.26 ng g^-1 wet weight, and expressed µ-opiate receptor as in expected size of 441 bp. According to our findings, it was concluded that T. canis, benefits using morphine-like substance to modulate host immunity.

Abstract This study was carried out to determine the presence and frequency of Anaplasma ovis and Anaplasma marginale in sheep and dairy cattle in West-Azerbaijan province, Iran. A total number of 200 blood samples were randomly collected via the jugular vein from apparently healthy cattle (100) and sheep (100). The extracted DNA from blood cells was screened using genus-specific (Anaplasma spp.) nested-polymerase chain reaction (PCR) based on 16S rRNA gene primer sets. Species-specific PCR was set up using major surface protein 4 (MSP4) gene primer set. None of cattle blood samples were positive for Anaplasma spp. by the first nested PCR. Five samples among the 100 sheep blood samples were both positive in the first nested PCR and A. ovis -specific PCR, based on MSP4 gene. In total, 5.00% of animals were A. ovis positive. This study identified a low prevalence of A. ovis in the blood of apparently healthy sheep in West Azerbaijan province.

Abstract Dog is the main reservoir host of visceral leishmaniasis in Iran. The present study was carried out to investigate visceral leishmaniasis in owned dogs with dermal lesions in Mashhad, Khorasan Razavi province. Thirty- nine owned dogs with dermal lesions were selected. During study, four of dogs were euthanized. The dermal smears, blood and tissue samples were collected and examined using parasitological, serological and molecular methods. A total of 39 examined dogs, leishman bodies were microscopically detected in 33.30% (13/39) of dermal smears. The sera samples were tested by indirect immunofluorescent antibody test (IFAT). Antibody against Leishmania infantum was detected in 26.00% (10/39) dogs. According to semi-nested PCR, DNA of Leishmania infantum was detected in 2.50% (1/39) of blood samples and in 75.00 % (3/4) of different tissues of euthanized dogs. BLAST analysis of the sequenced samples indicated a 99.00% similarity with kDNA of Leishmania infantum. Based on the results, it is concluded that visceral leishmaniasis due to L. infantum is distributed among household dogs of this area and it needs more surveillance to control the disease by public health and veterinary authorities.

Abstract The present study was conducted to evaluate the status of the parasite fauna in Acipenser persicus at different development stages, in order to find prevention protocols for parasitic diseases in this valuable species. For this purpose, sampling from each sex breeder, 10 egg samples, 5-day-old larvae (n = 20), 20-day-old larvae (n = 80) and fingerling of A. persicus (n = 60) released in earthen ponds were done. After the bioassay and preparing wet mount from the internal and external organs, identification was done according to the keys. According to the results, no fauna parasites were isolated from egg samples and 5-day-old larvae; but Trichodina spp. was isolated from 20-day-old larvae. Also, the same protozoan was isolated from fingerling released in earthen ponds, the mean intensity, prevalence and range of contamination by fingerling were higher with compared to 20-day-old larvae. Trichodina sp. and Diplostomum spathaceum were isolated from skin and eyes of females, respectively. However, Trichodina sp. and Ichthyophthirius multifiliis were isolated from skin of male breeders. In this study, no parasites were isolated from internal organs of larves and fingerling but four intestinal parasites included: Cucullanus sphaerocephlaus, Anisakis sp., Skyrjabinopsilus semiarmatus, and Lepto-rhynchoides plagicephalu were isolated from internal organs of breeder. Based on a wide range of parasitic infection observed in various life stages of A. persicus, it seems necessary to consider hygienic and management measures.

Abstract From April 2009 to December 2011, 44 dead hedgehogs (Erinaceus europaeus) were collected incidentally from areas of Urmia, Iran. The overall prevalence of helminth infections was 95.0%. Specific parasites and their prevalences were: Physalopteraclausa(93.0%), Crenosoma striatum (61.0%), Capillariaaerophila(9.0%), Capillariasspp.(4.0%), Brachylaemuserinacei(2.0%) and Hymenolepiserinacei(16.0%). There were no significant differences in helminth occurrence between hedgehog sexes, either in single or in mixed infections (p > 0.05). The mixed infection involving Crenosoma striatum and P. clausaoccurred significantly more frequently than other mixed infection (p < 0.05).There were significant differences in prevalence among seasons, with the highest prevalence in summer and spring especially among P. clausaand C. striatum (p < 0.05).

Abstract Forty-one wild sheep (Ovis ammon orintalis) from Kabodan Island of National Park of Urmia Lake (North-West of Iran), were examined during a period of six months from October 2002 to March 2003, for helminthes and coccidian infection. The numbers of oocyst and eggs per gram of faeces (OPG & EPG) were determined by the centrifuge flotation technique using saturated sugar solution. The rate of infection for Strongylid form, Marshalagia, Trichuris eggs, and lung worm larvae were 8 (19.5%), 12 (29.5%), 17 (41.5%) and 14 (34.1%), respectively. Thirty-three (80.48%) of the examined wild animals were infected to one or more Eimeria species including E. parva, E. ahsata, E. ovinoidalis and E. faurei. This study suggested that the rate of parasitic infection in wild sheep were very low but it would seem that in unsuitable condition such as drought and starvation, parasitic infection can be cause a serious problem in wild sheep population.