Establishment of primary cystic echinococcosis in laboratory mice: our results in the Balb/c strain.

Background: Establishment of experimental cystic echinococcosis (CE) in laboratory mice is required for the study of CE. Experimental CE may be established in primary or secondary forms; however, the primary form is more reliable for the study of CE. Aims: The aim of the study was to assess the possibility of establishment of primary CE in Balb/c mice via the oral administration of Echinococcus granulosus s.l. eggs. Methods: E. granulosus s.s. eggs were obtained from the gravid segments of the adult worms collected from an experimentally infected dog. Thirty-four female Balb/c mice were allocated into 4 groups and were orally infected with E. granulosus s.s. eggs as follows: 1) the mice with normal immunity, a single dose of 1000 eggs, 2) the mice with normal immunity, 3 doses of 500 eggs, 3) immunosuppressed mice, a single dose of 1000 eggs, and 4) immunosuppressed mice, 3 doses of 500 eggs. After 6.5 months, all the mice were opened and their internal organs were examined for the presence of CE cysts. The livers of infected mice were also examined for the presence of E. granulosus s.l. compartments by the PCR method. Results: There was no developed or developing CE cyst in the abdominal cavities or on the internal organs of all the mice in four groups. In addition, in the molecular study, all the examined liver samples were negative for the parasite material. Conclusion: The results of the present study revealed that Balb/c mice are not a suitable host for establishment of primary CE following the oral administration of E. granulosus s.s. eggs.

Morphological and molecular analyses of the spiruroid nematode, Falcaustra araxiana Massino, 1924 (= Spironoura araxiana) from the European pond turtle (Emys orbicularis).

There is little information on the phylogenetic position and life cycle of family Kathlaniidae. Falcaustra araxiana is a member of this family which infects the large intestine of the European pond turtle (Emys orbicularis). In the present study, morphological data and molecular analyses based on the 18S rDNA were performed on different types of F. araxiana originating from the large intestine and gastric nodules in the turtle. Morphological data revealed both larvae and adult stages in the gastric nodules. In addition, all nematodes recovered in the large intestine were adult worms. GenBank accession numbers KM200715 and KM200716 were provided for adult F. araxiana located in the intestine and stomach, respectively, of E. orbicularis. The results of sequencing proved that these two types are completely similar. Accordingly, it can be hypothesized that nodule formation is a part of the life cycle of the parasite or a survival strategy. Furthermore, F. araxiana develops to the adult stage in the gastric mucosa prior to migrating to the large intestine. Phylogenetic analysis revealed that F. araxiana unexpectedly branched away from other members of the superfamily Seuratoidea (Truttaedacnitis truttae, Cucullanus robustus and C. baylisi) and showed a closer relationship with Paraquimperia africana, a member of the Ascaridoidea. It seems that phylogenetic reconstruction for the present parasite needs more detailed morphology, life cycle and molecular studies.

Dynamics of the enzymatic antioxidants during experimental caprine coccidiosis.

Caprine coccidiosis, caused by coccidian parasites from genus Eimeria, is considered as one of the most common and significant diseases in goats worldwide. The aim of this experiment was to investigate the responses of the enzymatic antioxidant systems during experimental coccidiosis. For this purpose, 20 newborn kids were selected. Ten were infected with sporulated oocysts of the most pathogenic species of Eimeria, and the remainder served as the control. Blood samples were taken at 0 (before inoculation), 3, 7, 14, 21, 28, and 35 days post-infection (dpi), and antioxidant-oxidant-related parameters were measured. Our data showed that the activities of the main erythrocyte antioxidant enzymes revealed significant declines at 7 dpi. These decreases were more evident at 14 to 21 dpi and then gradually enhanced to the normal values until 35 dpi; however, total antioxidant capacity revealed a remarkable decrease at 7 dpi and remained on the same level toward the end of the study. By contrast, serum levels of malondialdehyde (a biomarker of lipid peroxidation) and total homocysteine significantly increased at 21 and 14, 21, and 28 dpi, respectively. These observations suggest that caprine coccidiosis can impair the major antioxidant systems leading to remarkable oxidative damages during the infection. Furthermore, oxidative injuries could have a considerable linkage to the pathogenesis of Eimeria parasites.

Acute phase response in cattle infected with Anaplasma marginale.

This study was undertaken to evaluate the acute phase responses via the assessment of the concentration of serum sialic acids (total, lipid bound and protein bound), inflammatory mediators (IFN-γ and TNF-α) and acute phase proteins (Hp and SAA) in 20 adult crossbred cattle naturally infected by Anaplasma marginale. The infected animals were divided into 2 subgroups on the basis of parasitemia rate (<20% and >20%). Also, as a control group, 10 clinically healthy cattle from the same farms were sampled. Our data revealed significant decreases in red blood cell count (RBC), hematocrite (PCV) and hemoglobine (Hb) in infected cattle compared to healthy ones. Conversely, the concentrations of Hp, SAA, ceruloplasmin, fibrinogen, serum sialic acids and the circulatory IFN-γ and TNF-α were increased in the diseased cattle (P<0.05). In addition, it was evident that the progression of parasitemia in infected cattle did not induce any significant alterations in the hematological indices (RBCs, PCV and Hb) and the concentrations of Hp, SAA, ceruloplasmin and fibrinogen. SAA was the most sensitive factor to change in the diseased cattle. Therefore, increase in SAA concentration may be a good indicator of inflammatory process in cattle naturally infected with Anaplasma marginale.

Alterations of erythrocyte antioxidant mechanisms: antioxidant enzymes, lipid peroxidation and serum trace elements associated with anemia in bovine tropical theileriosis.

In order to investigate the alterations of erythrocyte protective antioxidant mechanisms, lipid peroxidation and trace elements associated with anemia in bovine tropical theileriosis, an infected group comprised of 50 crossbred Holstein cattle, about 1-2 years old, naturally infected with Theileria annulata, were divided into 4 subgroups according to their parasitemia rates (<1%, 1-3%, 3-5%, >5%) and also 10 healthy cattle as control were selected. Blood samples were taken and hematological parameters, the activities of antioxidant enzymes including superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase and serum concentrations of some antioxidant trace elements (copper, iron, zinc, manganese and selenium) were measured. As an index of lipid peroxidation, the level of Malondialdehyde (MDA) was also determined. The results showed a conspicuous decrease in the activities of SOD, GPX and catalase (P<0.01), and a significant decrease in the serum concentrations of Cu, Zn, Mn and Se in cattle with higher than 1% parasitemia (P<0.05) compared to the control. In addition, remarkable elevations in the MDA level (P<0.01) and serum concentration of iron (P<0.05) were observed in the infected animals. These findings pointed to the occurrence of exacerbating oxidative injuries to erythrocytes during parasitemia. Furthermore, it can be concluded that infection with T. annulata can interfere with protective antioxidant mechanisms of RBCs against oxidative damages, which promote the development of anemia.

Evaluation of erythrocyte antioxidant mechanisms: antioxidant enzymes, lipid peroxidation, and serum trace elements associated with progressive anemia in ovine malignant theileriosis.

Ovine malignant theileriosis is a fatal disease that is characterized by severe progressive anemia. In order to elucidate the underlying mechanisms involved in anemia, this study was designed to assess the antioxidant status and erythrocyte oxidative injuries in Iranian fat-tailed sheep that suffered from malignant theileriosis. The infected animals (infected group), composed of 50 Iranian sheep about 1-2 years old, naturally infected with Theileria sp., were divided into three subgroups according to parasitemia rates (<1%, 1-3%, 3-5%), and ten non-infected animals were also selected as the control group. Blood samples were taken and hematological parameters, the activities of antioxidant enzymes including superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase, erythrocyte osmotic fragility, and serum concentrations of some trace elements (copper, iron, zinc, manganese, and selenium), were measured. As an index of lipid peroxidation, the level of malondialdehyde (MDA) was also determined. According to the results, a significant decrease in red blood cell (RBC) count, packed cell volume, the activities of SOD, GPX, and catalase (P <0.001), and also serum concentrations of Cu, Zn, Mn, and Se (P < 0.05) were evident in the infected sheep. In contrast, significantly increased levels of MDA and erythrocyte osmotic fragility (P < 0.001) as well as serum concentration of iron (P < 0.05) were recorded in the infected animals. The significant decrease in antioxidant enzyme activities and substantial elevated levels of lipid peroxidation and erythrocyte osmotic fragility associated with the increase in parasitemia indicate increased exposure of RBCs to oxidative damage. Also, it appears that disturbed antioxidant defense mechanisms can promote the development of anemia in ovine theileriosis.

Eimeria species in wild rabbits (Oryctolagus cuniculus) in Fars province, Iran.

This investigation was accomplished during February to November 2008. A total of 71 wild rabbits (Oryctolagus cuniculus) of about 5 to 12 months age were collected alive from different parts of Fars province, south of Iran. Faecal sampling was carried out directly from recti and the oocysts were isolated using sedimentation and floatation techniques and the sporulated oocyst were identified based on morphological and biological characteristics. All the rabbits were apparently healthy and showed no clinical symptoms. Twenty two rabbits (31.0%) were positive for infection with Eimeria and six species including Eimeria perforans (18.3%), Eimeria magna (16.9%), Eimeria media (14.1), Eimeria irresidua (11.2%), Eimeria flavescens (4.2%), and Eimeria coecicola (2.8%) were identified. Eighty six percent of the infected rabbits showed mixed infections with two or three Eimeria species. Lack of clinical signs could be due to the agro ecological and environmental conditions of rabbit habitats specifically dry and hot climatic features in recent years. In addition, immunity induced by long term exposure to low doses of oocysts shedded by the carrier animals probably have pivotal role in impairing parasitic developmental cycles and preventing acute coccidiosis.