Neosporosis is an infectious disease primarily of cattle and dogs, caused by intracellular parasite, Neospora caninum. Neosporosis appears to be a major cause of abortion in dairy cattle worldwide and causes to huge economic loss to dairy industry. Therefore its diagnosis is necessary. The aim of the present study was to develop an in-house enzyme-linked immunosorbent assay (ELISA) and compare it with its available commercial diagnostic kit. A total of 280 cattle sere were collected from Ahvaz slaughterhouse. Indirect haemagglutination (IHA) and modified agglutination tests (MAT) were used to screen positive and negative sera. After achieving 87 positive and negative sera, commercial ID. Vet ELISA kit was used. Based on whole tachyzoite antigens (Nc, Np1 and Np2), in- house ELISA was designed and used for 87 mentioned sera. Sensitivity, specificity, predictive values and cut- offs were calculated. Mc Nemar′s analyzing was used for detection the agreement of in- house and commercial ELISA methods. Kappa statistical analysis was considered. Based on the results out of the 280 samples, MAT method detected N. caninum infection in 87 (31.07%) cases. There was not significant difference between the results obtained by in- house based on Nc antigen and commercial ELISA kits. A proper agreement was between two kits.

Many infections can infect genitalia and can affect the quality of fertility. Neospora is one of the protozoans that can infect the male and female's reproduction system. But in available investigations, relation between Neospora infection and hormones was not obtained. Therefore, the present study was aimed to investigate relation of Neospora infection and testosterone and thyroid hormone values in acute and chronic neosporosis. To achieve bulls with neosporosis, N. caninum agglutination test (NAT), ELISA and PCR were performed. Testosterone, thyroxin (T4) and tri-iodothyronine (T3) levels were measured and compared between infected and non-infected bulls. Results of the present study showed that testosterone value was decreased significantly in bulls with acute neosporosis. Serum T4 of bulls with chronic neosporosis was decreased significantly but changes of serum T3 was not significant. The hypothalamic–pituitary–adrenal axis (HPA axis) impairment, direct effect of N. caninum and its antigens on testis function and occurrence of oxidative stress in testis due to the presence of Neospora may be the reasons of hormonal changes.

Keywords: Haemoproteus, Pigeon, nested PCR, Histopathology
Pigeons have been used for a long time as a food resource, pets or cultural and religious symbols. Pigeons act as reservoir or carrier and an important source of infection for other avian hosts. Haemoproteus is pathogenic protozoan that effecting blood circulatory system of pigeons.
The present study was undertaken to evaluate the presence of Haemoproteus prevalence in pigeons from Ahvaz and its histopathological changes. A total of 108 blood samples were taken from pigeons to investigate Haemoproteus presence by molecular and blood smear methods. For molecular detection of Haemoproteus semi-nested PCR targeting the cytochrome b gene was used. Also to evaluate histopathological changes 14 infected pigeons to Haemoproteus were sacrificed and studied. Results of histopathological study showed that focal lymphocytic aggregates, pigmentation and vacuolation in livers, multifocal non-suppurative interstitial nephritis in kidneys, pigmentation and white pulp hyperplasia in spleens were the main histopathological lesions in infected pigeons. Moderate lymphocytic myocarditis was finding in heart of one pigeon. No histopathological changes were seen in brain, cerebellum, intestine and pancreas.
In conclusion, Haemoproteus is prevalent in pigeons from Ahvaz. Based on the parasites histopathological effects, more attention should be considered to Haemoproteus.
 

  Polyamines such as putrescine, spermidine, and spermine are ubiquitous in all eukaryotic cells and play an essential role in cell division and differentiation. One way of Polyamine biosynthesis is done by ornithine decarboxylase (ODC) which catalyzes the transformation of ornithine to putrescine. The aim of present study was to evaluate the level of putrescine, spermidine and spermin in protoscolices, hydatid fluid and germinal layer and to evaluate indirect ODC activity. In the present study putrescine, spermidine and spermin levels were investigated in germinal layers, hydatid fluids and protoscolices incubated with or without ornithine. The samples were homogenized and liquid chromatographic (HPLC) was used for polyamines measurement. Based on the results putrescine was the lowest polyamine and since its level was not increased in protoscolices incubated with ornithine, ODC activity was not detected. Spermidin was the highest polyamine and the results showed that germinal layer contained the highest level of polyamines.Overall, the results showed that probably ODC activity was not detected or at the low level in hydatid cyst and since germinal layers contained rapidly proliferating cells level of polyamines were higher than other parts. To confirm the obtained results more investigations are necessary

Leishmaniasis is protozoan disease that considered as one of the problems in public health. Due to toxicity and resistance of available drugs, development of new compounds is important. The aim of the present study was to evaluate in vitro effect of Scrophularia striata essential oil on Leishmania. tropica and Leishmania. major. First, essential oil was prepared and analysed with GC/MS. After exposure of parasites to different concentrations of essential oil, viability of promastigotes and axenic amastigotes were investigated by counting and colorimetric assay and the obtained results were expressed as 50% inhibitory concentration. Essential oil analysis showed that nonane, linalool and α-terpineol were the most abundant compounds. In the present study, by counting IC50 of S. striata was achieved 6.5 and 1.5% for L. tropica promastigotes and axenic amastigotes, and 8 and 5% for L. major, respectively. In MTT assay IC50 was obtained 5.8 and 2.1% for promastigotes and axenic amastigotes of L. tropica and 6 and 4.5% for L. major, respectively. Overall, according to S. striata low cost, availability and acceptable leishmanicidal effects, further investigation can be done.

 Many infections can infect genitalia and can affect the quality of fertility. Neospora is one of the protozoans that can infect the male and female's reproduction system. But in searches, relation between Neospora infection and sperm parameters was not observed. Therefore, the present study with the aim of investigation on relation of Neospora infection and sperm parameters in acute and chronic neosporosis were performed. After achieving 15 bulls with acute neosporosis diagnosed by PCR, 15 bulls with chronic neosporosis diagnosed by modified agglutination test and 15 healthy bulls, some sperm specifications such as sperm count, viability, motility and morphology were studied and compared in different groups. Results of the present study showed that sperm count; viability and somewhat motility had significant relation with neosporosis. SOD activity in acute neosporosis was higher than other groups but the difference was not significant. Also, although malondialdehyde was higher in acute group but the difference was not significant. Overall, the present study showed that probably some of the factors related to sperm health could be affected with Neospora infection.

Equine theileriosis is one of the most important parasitic diseases among the equids that poses a serious threat to the horse industry and has important implications for the international movement of horses. Proper diagnosis of parasites in carriers is important in for prevention and effective control measures. Therefore, the aim of the present study was to investigate probable presence of Theileria equi infection in dogs from rural areas around Ahvaz.
A total of 100 blood samples from dogs were examined for the presence of T. equi infection using molecular methods. For molecular detection of T. equi, primers targeting the 18SrRNA gene were selected. After DNA extraction from the dog's whole blood, PCR was performed and the results were observed in agarose gel. Out of 100 samples, the PCR method detected T. equi infection in two dogs. In other words two percent of dogs were carriers for T. equi.
In conclusion, the present study revealed that dogs from rural areas around Ahvaz are infected with T. equi. So these animals probably can be considered as the carriers of T. equi. Result of this study propose that dog's infection with T. equi should be mentioned in equine theileriosis control programs.
 

 

Abstract
Surname: Azadmanesh Name: Somayeh
Title: Antioxidant status during the course of acute and chronic toxoplasmosis in rat
Supervisor/s: Dr. Somaye Bahrami, Dr. Ali shahriari
Advisor/s:: Dr. Mehdi Tavalla
Degree: Doctor of Veterinary Medicine University: Shahid Chamran University of Ahvaz, Iran
Faculty: Veterinary Medicine Department: Pathobiology
Keywords: Oxidative stress; Rat; Toxoplasma gondii .
Toxoplasmosis is a common parasitic infection in the world. During pregnancy, especially in the animal cases, toxoplasmosis cause severe fetal malformation. Since, increased free radicals and oxidative stress are reported in many parasitic diseases the purpose of the present study was to evaluate the oxidative stress in acute and chronic toxoplasmosis. RH strain of Toxoplasma tachyzoites were used in the present study. Twenty five rats were infected with the parasite while 25 other rats were as the control group that received normal saline. 0, 5, 7, 10 and 45 days post infection blood samples were taken. Some parameters related to oxidant and antioxidants such as malondialdehyde, catalase, total antioxidant capacity, glthatione and superoxide dismutase were measured. Results were analyzed using appropriate statistical methods. In seventh (p=0.019) and tenth day (p=0.027) the amount of glutathione was significantly different between the infected and non-infected group. Also, the amount of total antioxidant capacity was significantly reduced in seventh day. In other cases, there was no significant difference between the groups in different days. Overall, based on the results it seems that, in the seventh day, in infected rats responses to oxidative stress were triggered and led to decrease of total antioxidant capacity. Furthermore, glutathione was increased to cope with stress. It seems that antioxidant defense system entered the infection to the chronic phase and changed the parasites stage.

 

 Neospora caninum is a protozoan parasite that presents worldwide distribution and is mainly implicated as responsible for bovine abortion. There are indications that the presence of birds in cattle raising farms could be associated with the increase of seroprevalence and abortions in related to N. caninum. Although the presence of birds in cattle-raising properties is positively correlated to higher infection rates, little has been described about the role of these animals in the parasite's life cycle. In that sense, the aim of this study was to determine the frequency of N. caninum infection in sparrows using molecular tests. A total of 210 sparrows were included in this study. For molecular study, DNA was extracted using a genomic DNA purification kit and for detection of N. caninum, primers targeting Nc5 gene were selected and PCR was conducted. The results showed that 6 (2.8%) of brain samples were positive for N. caninum. Sequenced samples were found to be closest to the N. caninum Nc 5 gene with a similarity of ≥98%. The results that obtained in the present study indicated soil contamination due to N. caninum oocysts because sparrows feed from the ground, and suggested that the meat from the sparrows might be an important source for dog infection by N. caninum. The present findings could indicate a role for sparrows in the epidemiology of neosporosis. However detection and/or isolation of the parasite in tissues would be necessary to corroborate that sparrows are intermediate hosts of N. caninum

Recently, herbal medicines have increasingly been used to treat many diseases, including several infections. Since Lepidium sativum has been shown to have a number of medicinal properties, in this study the scolicidal effects of the essential oil (EO) from this plant was investigated. Lepidium EO was obtained by hydrodistillation method. Gas chromatography-mass spectrometry (GS-MS) were employed to determine the chemical composition of the EO. Protoscolices were exposed to various concentrations of EO (1, 3, 5, 10 and 15 mg/ml) for 10, 20, 30 and 60 min. Viability of protoscolices were confirmed by 0.1% eosin staining.
A total of 19 compounds representing 95.5% of the total oil, were identified. α-Thujene (88.86%), Myrcene (2.9%) and P-cymene (1.67%) were found to be the major EO constituents. The mortality rate of protoscolices at 1 mg/ml of Lepidium was 8.3, 9.4, 12.3 and 18.8% after 10, 20, 30 and 60 min, respectively. Scolicidal power of EO at 1 mg/ml was 24.2, 24.8, 25.7 and 37.1% after mentioned incubation times, respectively. The EO at concentration of 5 mg/ml killed 49.2, 56.1, 77.4 and 93.5% after 10, 20, 30 and 60 min, respectively. After 10 and 20 mins, 10 mg/ml of EO killed 88.64 and 96.2% of protoscolices. One hundred percent scolicidal activity was observed with Lepidium EO at concentration of 10mg/ml after 30 min of exposure.
The results of present study revealed that the EO of Lepidium is rich in α-Thujene and has high scolicidal power. This plant may be used as a natural scolicidal agent.
 

<p>&lt;p&gt;&amp;lt;p&amp;gt;Leishmaniasis is parasitic disease that is an important problem of public health worldwide. Intramuscularly administered glucantime and pentostam are the most common drugs used for treatment of the disease, but they have significant limitations due to toxicity and increasing resistance. Therefore, development of new chemotherapeutic agents are important for the control of disease. The aim of this study was to access the cytotoxic effect of amiodarone on Leishmaniamajor and L. tropica promastigotes and to observe the programmed cell death features. The colorimetric assay was used to find L. major and L. tropica viability and the obtained results were expressed as 50% inhibitory concentration. Annexin -V FLUOS staining was performed to study the cell death properties of amiodarone using FACS analysis. Qualitative analysis of the total genomic DNA fragmentation was performed by agarose gel electrophoresis. Furthermore, to observe changes in cell morphology, promastigotes were examined using light microscopy. The IC50 was achieved at 81 &amp;amp;micro;M and 55 &amp;amp;micro;M for L. major and L. tropica after 48 hr of incubation, respectively. In both strains of L. major and L. tropica, amiodarone induced death with features of apoptosis, including cell shrinkage, DNA fragmentation, and externalization of phosphatidylserine. Our results indicate that amiodarone induces apoptosis of the causative agent of cutaneous leishmaniasis.&amp;lt;/p&amp;gt;&lt;/p&gt;</p>

: Neospora caninum is a protozoan parasite that presents worldwide distribution and is mainly implicated as responsible for bovine abortion. There are indications that the presence of birds in cattle raising farms could be associated with the increase of seroprevalence and abortions in related to N. caninum. Although the presence of birds in cattle-raising properties is positively correlated to higher infection rates, little has been described about the role of these animals in the parasite's life cycle. In that sense, the aim of this study was to determine the frequency of N. caninum infection in pigeons using serologic and molecular tests. Materials and Methods: A total of 102 pigeons were included in this study. Samples of blood were collected and sera were separated. Brain tissues were used for DNA extraction. Neospora caninum agglutination test (NAT) was performed according to the method previously described for toxoplasmosis. For molecular study, DNA was extracted using a genomic DNA purification kit and for detection of N. caninum, primers targeting Nc5 gene were selected and PCR was conducted. Results: Antibodies to N. caninum were found in 30.39% (31/102) using agglutination test, with titers ranging from 1: 4 to 1: 128. Neospora DNA was detected in 10/102 (9.8%) pigeons. Discussion: The results that obtained in the present study indicated soil contamination due to N. caninum oocysts because pigeons feed from the ground, and suggested that the meat from the pigeons might be an important source for dog infection by N. caninum. The present findings could indicate a role for pigeons in the epidemiology of neosporosis. However detection and/or isolation of the parasite in tissues would be necessary to corroborate that pigeons are intermediate hosts of N. caninum.

In the history, piroplasmids were generally considered as specific parasites in different animals. The specificity of piroplasmids is probably lower than expected, which is supported by detection of non-specific species from Human and another animals. According to the importance of piroplasmids infection in raising industry and international movement of camels in different country specially Iran, the aim of the present study was to characterize species of camels piroplasmosis agents in the original regions of camel raising of Iran. Out of the 248 samples, the PCR method detected Theileria equi and Babesia caballi infection in 16 (6.45%) camels. Sequences of 18S rRNAs from all isolates had more than 99% similarity to each other and 96% to B. caballi and T. equi isolated from camels from other studies. Based on single nucleotide substitution in 18S rRNA gene of studied camels, 3 different genotypes of B. caballi and 7 different genotypes of T. equi were identified and submitted to GeneBank. Based on the homologies between 18S rRNA sequences of B. caballi and T. equi obtained from studied camels and those submitted to the GeneBank, 2 phylogenetic tree formed two and three distinct highly related clusters respectively. Age, gender and localities were not determined as risk factors for T. equi and B. caballi infection in camels in this study. In conclusion, this study demonstrated that camels of Iran are infected by T. equi and B. caballi.

<p>: Neospora caninum is protozoan parasite which can cause a serious disease in dogs and cattle. It has been shown that birds may be a permissive intermediate host for N. caninum since parasite DNA has been detected in tissues from birds. It is showed that embryonated chicken egg can be used as an animal model for experimental infection. The aim of present study was to compare experimental infection of Neospora in chicken and pigeons embryonated eggs.<br /> Methods: An infection with N. caninum Nc1 isolate was conducted in chicken and pigeons embryonated eggs to evaluate LD50. After calculation of LD50, 2LD50 of tachyzoites were injected to eggs. Macroscopic changes of each embryo were noticed and to investigate the parasite distribution in tissues immunohistochemistry (IHC) and molecular methods were used. In the present study, histopathological changes were considered and sections to those used for histopathological examination including heart, liver, brain and chorioallantoic (CA) membrane were subjected to IHC, too. For PCR procedure, primer pair Np21/Np6 was used for amplification of the Nc5 gene.<br /> Results: Pigeon's embryo showed more macroscopic changes than chicken embryo. A hemorrhage of the CA was the main grass lesion. All the infected tissues had histopathological changes. Microscopic examination of tissues revealed acute neosporosis due to hemorrhage, necrosis and infiltration of mononuclear inflammatory cells. Based on IHC and molecular results, the parasite aggregation in the heart was more predominant than in the other tissues. <br /> Discussion: These results reinforce that there is genetic susceptibility to N. caninum in pigeons embryonated eggs like chickens embryonated eggs and provide new insights to research an inexpensive and available animal model for N. caninum.<br /> &nbsp;</p>

 Introduction: Equine theileriosis poses a serious threat to the horse raising industry and international movement of horses. Since carrier animals are the major sources for spreading the infection, therefore their identification is necessary. Objective: The aim of the present study was to investigate the presence of T. equi in camels as the carrier animals in Yazd province where there is a considerable population of camels and horses and equine theileriosis is prevalent. Materials and Methods: In this study, blood samples were taken from 161 camels from different regions of Yazd province. For identification of T. equi PCR was used with amplification of 18 S rRNA gene. Positive samples were sequenced. Result: Out of the 161 samples, the PCR method detected T. equi infection in 7 (4.3%) camels. Sequences of 18S rRNAs from all isolates had more than 99% similarity to each other and to T. equi isolated from horse and camels from other studies. Based on single nucleotide substitution in 18SrRNA gene of studied camels, 3 different genotypes were identified and submitted to GeneBank. Discussion: Based on the homologies between 18SrRNA sequences of T. equi obtained from studied camels and horses and those submitted to the GeneBank the phylogenetic tree formed three distinct highly related clusters. Age, gender and localities were not determined as risk factors for T. equi infection in camels in this study. In conclusion, this study demonstrated that T. equi is present in the camels of Iran and camels should be considered as the carriers for T. equi

Introduction: Antileishmanial drugs that are traditionally used for treatment of Cutaneous leishmaniasis are mainly toxic, ineffective for some parasite isolates, and mostly expensive. Currently, more attention is focusing on natural active components. The increased awareness of the potentials and industrial value of chitosan biopolymer lead to its utilization in many applications such as in the medical and veterinary arenas. In this study, we aimed to clarify assessing the effectiveness of nanochitosan films in the treatment of cutaneous leishmaniasis caused by L. major (Iranian strain). Materials and methods 0.1 ml of the solution containing 2×106 L. major promastigotes (MRHO/IR/75/ER) was injected into the base of 36 mice tail subcutaneously. After 6wk of inoculation, when lesions appeared at the base of the tail, treatment was initiated. Animals were divided into 4 groups: Negative control group that parasites were injected without any treatment, Positive control group treated with daily peritoneal injection of glucantim, Treatment group 1 which treated with nanochitosan film, Treatment group 2 which treated with nanochitosan film and with daily peritoneal injection of glucantim. In each group mice were equally euthanized after seven, fourteen and twenty one day and the lesion size and parasite load were evaluated. Also, tissue samples were obtained from the wound area and the repair process was examined microscopically. Results: Both nanochitosan film and glucantime produced suppression and reduction effect in the size of lesions compare with control group. There was no significant difference between nanochitosan and glucantime in reduction of lesion size. But lesion size in the group that treated with combination of nanochitosan film and glucantime was significantly reduced from two weeks. Significant reduction in the parasite load was found in the group of mice treated with combination of nanochitosan film and glucantime. Wound healing process occurred in group which treated with nanochitosan film, glucantime and combination of nanochitosan film and daily peritoneal injection of glucantim. Conclusion: The injection of glucantim can increase the wound healing effect of nanochitosan film. Therefore, their combination probably can be used for treatment of cutaneous leishmaniasis.

Equine theileriosis (ET) is a febrile, tick-borne disease caused by Theileria equi (T. equi). The etiological agent of ET is specific for solipeds. The disease poses a serious threat to the horse raising industry and international movement of horses. Based on our knowledge, there is no previous study regarding the prevalence of T. equi infections in Iran. In this study, we aimed to determine the presence of parasite DNA in the equine population in Khuzestan province, southwest of Iran. A total of 165 blood samples from healthy horses from different stables in Khuzestan province, southwest of Iran, were examined for the presence of Theileria equi infection using parasitological and molecular methods. Out of the 165 samples, the parasitological method detected T. equi infection in no cases while the PCR method gave 47 (28.48%) positive results. Age, gender, kind of activity, contact with other animals and breed were not found as risk factors for T. equi infection in this study. Significant difference was identified in different geographic locations. Since blood samples were taken from healthy animals, this implies that 28.48% of horses had subclinical theileriosis in the current study. In conclusion, this study demonstrated that T. equi is present in horses in the southwest of Iran. Despite the healthy appearance of horses, these carrier animals can transmit the parasites to ticks and are a potential continuous source for maintaining and disseminating the organisms to the horse population. We concluded that it is important to make further studies on definitive host and vectors in the respective areas.