Prevalence of Neospora caninum as an etiologic agent of animal abortion in Kurdistan Region of Iraq.

Neospora caninum ( N. caninum) is the etiologic agent of neosporosis, a potential cause of severe reproductive disorders in cattle, small ruminants, equines, wild animals and canids across the world. The current study is performed to estimate molecular prevalence of N. caninum in small ruminants and equines that had abortion in Kurdistan region of Iraq. A total of 64 tissue samples (brain, placenta, heart, lung and liver) were taken from aborted foetuses, with a total of 122 dam blood samples taken from 63 sheep, 39 goats, 12 mares and 8 jennies in local breed fields. Besides, a risk factor analysis for N. caninum positive animals was performed. The observed prevalence of N. caninum DNA in the blood of sheep, goats, horses and donkeys were 20.6%, 17.9%, 21.4% and 25.0%, respectively, and 19.3%, 17.6%, 18.1 and 20.0% in the aborted foetuses of the animals, respectively. Moreover, occurrence of N. caninum was 20.3% in the blood of aborted dams, while it was 18.7% in their aborted foetuses. Confirmatory analysis was also done through constructing a phylogenetic tree to compare the partial sequences of the Nc-5 gene in our isolates (OP771519, OP771520, OP771521 and OP771522) with the GenBank sequences. This showed 98-100% sequence identity with other N. caninum strains in the GenBank database. Older small ruminants and equines had a higher risk of being positive for N. caninum and exposure to dogs were considered as significant risk factors for N. caninum infection in the studied animals (p<0.05). Thus, the results of this study suggest that N. caninum is one of the microbial abortive agents in small ruminants and equines in Kurdistan region of Iraq. It is hoped that the results of this study will help to control animal abortion in livestock and reduce the economic losses.

Phylogenetic Diversity of Dermanyssus gallinae (Dermanyssidae) based on Mitochondrial Cytochrome Oxidase-1 Gene Sequence Collected from Different Bird Species in Iran.

A wide range of hosts, especially birds, can be infested with Dermanyssus gallinae (D. gallinae), as an obligate hematophagous mite. In this study, cytochrome oxidase 1 (CO1) gene sequences were employed to perform molecular and phylogenetic analyses of D. gallinae collected from different bird species in Iran. Adult mites were collected from the body surface and cage material of ornamental and wild birds in industrial farms located in the Western and Northwestern regions of Iran. The infestation was identified in layer poultry farming by inspecting the eggs and the whole surfaces of the birds' bodies. The holding area and body surface of the ornamental and wild birds were also thoroughly examined. The D. gallinae samples were assigned to two subgroups of haplogroup A (i.e., A1 and A2). The phylogenetic tree suggested that the D. gallinae samples collected from wild birds in the A1 sub-haplogroup should be placed beside Japanese, Norwegian, Italian, and French samples isolated from wild birds in the A2 sub-haplogroup. Additionally, the highest phylogenetic similarity in the A2 sub-group was observed between mites isolated from ornamental and industrial birds in Australia. The findings of the present study suggest that crows and sparrows may play an important role in the transmission of D. gallinae infestation to other species of wild birds due to their high population, as well as their presence in most areas.

Toxicities of the copper and zinc oxide nanoparticles on Marshallagia marshalli (Nematoda: Trichostrongylidae): evidence on oxidative/nitrosative stress biomarkers, DNA damage and egg hatchability.

This study investigated the in vitro anthelmintic activity of copper oxide (CuO) and zinc oxide (ZnO) nanoparticles (NPs) against Marshallagia marshalli. The in vitro study was based on an egg hatch assay, adult and larvae motility inhibition assays, DNA damage, intensity protein profile along with several oxidative/nitrosative stress biomarkers including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), protein carbonylation (PCO), malondialdehyde (MDA), total antioxidant status (TAS) and nitric oxide (NO) content. Different concentrations of CuO-NPs and ZnO-NPs (1, 4, 8, 12 and 16 ppm) were used to assess anthelmintic effects on three stages of M. marshalli life cycle - that is, eggs, larvae and adult parasites for 24 h. The results indicated that CuO-NPs and ZnO-NPs played a significant role as anthelminthics, and the effect was dependent on time and concentration. The concentrations of 12 and 16 ppm of CuO-NPs and 16 ppm of ZnO-NPs resulted in the induction of oxidative/nitrosative stress (decreased SOD, GSH-Px and CAT, and increased MDA, PCO and NO), increased DNA damage, inhibition of adult and larval motility, egg hatch and low intensity of protein bands following sodium dodecyl sulphate-polyacrylamide gel electrophoresis, compared to control. It was concluded that CuO-NPs and ZnO-NPs could be utilized as novel and potential agents for the control and treatment of M. marshalli infection, and they have the pharmacological potential to be studied in vivo for further utilization in treating parasitic infections.

Molecular Characterization and Phylogenetic Analysis of Pathogenic Theileria spp. Isolated from Cattle and Sheep based on Cytochrome b Gene in Iran.

The present study investigated the phylogenetic relationship based on cytochrome b gene sequences among pathogenic Theileria species (spp.) in Iran, including Theileria annulata and Theileria lestoquardi, along with other data available in GenBank. A total of 136 (cattle) and 80 (sheep) blood samples suspected of piroplasm infection were obtained from six different provinces of Iran. Both microscopic and molecular methods using species-specific primers were used for screening T. annulata and T. lestoquardi positive samples. Finally, the partial cytochrome b gene of 30 T. annulata and 5 T .lestoquardi were amplified, sequenced, and deposited in GenBank. The results indicated that there were 12 different genotypes among T. annulata isolates, while only one genotype was observed among T. lestoquardi isolates. T. lestoquardi infection in cattle was detected in one sample, and no T. annulata and T. lestoquardi coinfection were detected in sheep and cattle. In the phylogenetic tree, different Theileria spp. were placed in separate clades, and the reliability of depicted tree and monophyly of T. annulata and T. lestoquardi ingroups were supported by the bootstrap value of 94% which significantly indicated that these two species evolved from a common ancestor. The tree also showed that these two pathogenic spp. shared a more recent common ancestor, compared to another species of Theileria parasites. To the best of our knowledge, this study is the first phylogenic analysis of pathogenic Theileria spp. in Iran based on the cytochrome b gene sequences. In addition, the first T. lestoquardi cytochrome b gene was sequenced and deposited in GenBank.

Assessment of oxidative/nitrosative stress biomarkers and DNA damage in Teladorsagia circumcincta following exposure to zinc oxide nanoparticles.

Drug resistance to helminth parasites is one of the most serious problems to threaten the livestock industry. The problem also poses a major threat to public health. Therefore, novel and safe agents should urgently be investigated to control parasitic infections. The current study was conducted to evaluate the possible antiparasitic effects of zinc oxide nanoparticles (ZnO-NPs) on one of the most prevalent gastrointestinal nematodes, Teladorsagia circumcincta. The worms were incubated with various concentrations of ZnO-NPs: 1, 4, 8, 12 and 16 ppm for 24 hours. Mobility and mortality of the parasites were recorded at four-hour intervals. At the endpoint, several biomarkers of oxidative/nitrosative stress, including superoxide dismutase, glutathione peroxidase and catalase, as well as lipid peroxidation, protein carbonylation, total antioxidant status, nitric oxide contents and DNA damage, were measured in the homogenized samples. ZnO-NPs showed significant anthelminthic effects, depending on time and concentration. Furthermore, the nanoparticle induced severe oxidative/nitrosative stress and DNA damage. ZnO-NPs could be considered as a novel and potent anthelminthic agent.

Histopathological changes and oxidative damage in hepatic tissue of rats experimentally infected with Babesia bigemina.

The present study was aimed to investigate oxidative stress, DNA damage, and histopatholog- ical alterations in hepatic tissues of splenectomized Wistar rats experimentally infected with Ba- besia bigemina. Rats were challenged with 5x106 infected erythrocytes. Babesia infection was con- firmed both with Giemsa's staining blood smears and nested-PCR amplified region of apical membrane antigen-1 (AMA-1) gene. Parasitemia reached approximately 10 % at day 5 post-in- fection. Livers of infected rats were enlarged and darker in color, became extremely brittle with marked congestion. Microscopic evaluation showed cytoplasmic clearing of hepatocytes and se- vere hydropic changes with significantly dilated sinusoids containing macrophages and also intra- sinosoidal parasitized erythrocytes. Severe infiltration of lymphoplasma cells was also present throughout the liver parenchyma. Furthermore, Kupffer cells were enlarged and, occasionally, containing Babesia-parasitized erythrocytes. The activity of Glutathione (GSH) and catalase (CAT), and total antioxidant capacity (TAC) were also significantly decreased (p ⟨ 0.05) after infection of rats with B. bigemina. B. bigemina infection also induced a significant increase (p ⟨ 0.05) in hepatic malondialdehyde (MDA) and nitric oxide-derived products (NOx) concentra- tions as well as amount of endogenous hepatocytes DNA damage. Hepatic damage was also re- flected through the measurement of lactic acid dehydrogenase (LDH) and protein carbonyl con- tent (PCO) in liver cells. These two indices of liver injury were also significantly elevated (p ⟨ 0.5) during B. bigemina infection. Evaluation of correlation between assayed variables in infected rats revealed that MDA levels were positively correlated with PCO, NOx, LDH and DNA damage in the infected group and negatively correlated with GSH, CAT and TAC. There was also an inverse relationship between the antioxidant enzymes activities of GSH, CAT and TAC with PCO, NOx and DNA damage in infected rats. However, NOx showed positive correlation with PCO and DNA damage in infected rats. On the basis of the above results it can be concluded that the Ba- besia infection increases oxidative stress markers, protein carbonyl content and DNA damage and decreases antioxidant enzymes activities in the liver. These results suggest that B. bigemina infec- tion could alter the liver histopathology and causes DNA damage following oxidative stress in hepatic tissue. Further studies are needed to precisely define how hepatic tissue damage takes place in B. bigemina infection.

Seroprevalence and Risk Factors of Neospora caninum and Toxoplasma gondii in Small Ruminants in Southwest of Iran.

Toxoplasma gondii and Neospora caninum are Apicomplexan intracellular protozoa with global distribution. Small ruminants play an important role as intermediate hosts for N. caninum and T. gondii, parasites of great public health concern. The main goal of the current survey was to evaluate N. caninum and T. gondii infection rate in sheep and goats of Khuzestan Province, southwest of Iran, using enzyme-linked immunosorbent assay (ELISA). In this cross-sectional study during February-April 2016, whole blood samples were taken randomly from 735 animals from 37 herds. The animals were reared under the traditional husbandry system in different parts of the province. Among 550 sheep and 185 goats, 37 (6.8%) sheep and 20 (10.8%) goats were seropositive for N. caninum and 59 (10.8%) sheep and 37 (20%) goats were seropositive for T. gondii. The incidence rates of mixed infection with N. caninum and T. gondii were 3.2% and 5.4% in sheep and goats, respectively. Seroprevalence rate of N. caninum was significantly higher in goats at T. gondii and abortion (18.2%). Also, a significant correlation was detected between seroprevalence of N. caninum and T. gondii and mixed infection in goats with a history of abortion. This is the first report of IgG antibody production against N. caninumand T. gondii co-infection in small ruminants in Iran. Our findings indicated that neosporosis and toxoplasmosis may be responsible for abortion in small ruminants in this region. Therefore, further investigations are needed to improve sanitary strategies in animals&rsquo; husbandry and launching control programs.