Molekularna i serološka istraživanja invazije vrstom Neospora caninum u golubova u jugozapadnom Iranu.

Neospora caninum is a protozoan parasite with worldwide distribution, mainly implicated as responsible for bovine abortion. There are indications that the presence of birds on cattle farms could be associated with the increase in seroprevalence and abortions related to N. caninum. The present study reports the serological (Neospora agglutination test) and molecular (PCR) presence of N. caninum in pigeons. From the 102 samples analyzed, 31 samples (30.39%) were seropositive for N. caninum and the overall molecular prevalence of N. caninum in the brains of the same pigeons was 9.8% (10/102). This is the first report of detection of N. caninum in Iranian pigeons. The results indicate soil contamination due to N. caninum oocysts, because pigeons feed from the ground, and suggest that the meat from the pigeons may be an important source for infection of dogs.Neospora caninum praživotinja je proširena diljem svijeta koja pretežito uzrokuje pobačaj u goveda. Upozorava se da prisutnost ptica na goveđim farmama može biti povezana s povećanjem serološke prevalencije i broja pobačaja uzrokovanih tom vrstom. U ovom je istraživanju N. caninum dokazana u golubova serološki testom aglutinacije i molekularno lančanom reakcijom polimerazom. Od 102 pretražena uzorka 31 (30,39%) je bio serološki pozitivan na prisutnost N. caninum, a molekularna prevalencija u mozgu istih golubova iznosila je 9,8% (10/102). To je prvo izvješće o dokazu N. caninum u iranskih golubova. Rezultati pokazuju da je onečišćenje tla oocistama N. caninum izvor uzročnika za golubove jer se oni hrane na tlu. Također se upozorava na činjenicu da meso golubova može biti važan izvor zaraze za pse

Molekularna i serološka istraživanja invazije vrstom Neospora caninum u golubova u jugozapadnom Iranu.

Neospora caninum is a protozoan parasite with worldwide distribution, mainly implicated as responsible for bovine abortion. There are indications that the presence of birds on cattle farms could be associated with the increase in seroprevalence and abortions related to N. caninum. The present study reports the serological (Neospora agglutination test) and molecular (PCR) presence of N. caninum in pigeons. From the 102 samples analyzed, 31 samples (30.39%) were seropositive for N. caninum and the overall molecular prevalence of N. caninum in the brains of the same pigeons was 9.8% (10/102). This is the first report of detection of N. caninum in Iranian pigeons. The results indicate soil contamination due to N. caninum oocysts, because pigeons feed from the ground, and suggest that the meat from the pigeons may be an important source for infection of dogs.Neospora caninum praživotinja je proširena diljem svijeta koja pretežito uzrokuje pobačaj u goveda. Upozorava se da prisutnost ptica na goveđim farmama može biti povezana s povećanjem serološke prevalencije i broja pobačaja uzrokovanih tom vrstom. U ovom je istraživanju N. caninum dokazana u golubova serološki testom aglutinacije i molekularno lančanom reakcijom polimerazom. Od 102 pretražena uzorka 31 (30,39%) je bio serološki pozitivan na prisutnost N. caninum, a molekularna prevalencija u mozgu istih golubova iznosila je 9,8% (10/102). To je prvo izvješće o dokazu N. caninum u iranskih golubova. Rezultati pokazuju da je onečišćenje tla oocistama N. caninum izvor uzročnika za golubove jer se oni hrane na tlu. Također se upozorava na činjenicu da meso golubova može biti važan izvor zaraze za pse

Development of an Indirect ELISA Using Different Fragments of Recombinant Ncgra7 for Detection of Neospora caninum Infection in Cattle and Water Buffalo

Background: Dense granules are immunodominant proteins for the standardi-zation of immunodiagnostic procedures to detect neosporosis. In the presented study different fragment of a dense-granule protein was evaluated for serodiagno-sis of Neospora caninum in cattle and water buffalo. Methods:NcGRA7, from N. caninum tachyzoites was amplified. PCR product and pMAL-c2X plasmid were digested with EcoR1 restriction enzyme and ex-pressed in Escherichia coli to evaluate its competence for detection of anti- N. cani-num antibodies with ELISA in comparison with commercial IDEXX ELISA. Fur-thermore, 230 sera of presumably healthy cattle and water buffaloes (108 cattle and 122 water buffaloes) were analyzed by both tests to determine the agreement of these two procedures. Results:Sensitivities and specificities of NcGRA7-based ELISA were 94.64% and 90.38% respectively using sera of cattle, but were 98.57% and 86.54% in the case of buffaloes respectively. A good correlation between the results of IDEXX ELISA and ELISA based on recombinant NcGRA7 for detecting N. caninum an-tibodies was appeared. Analyzing by Mc Nemar´s showed that NcGRA7-based ELISA has acceptable capability to differentiate the positive results in comparison with IDEXX ELISA. Conclusion:NcGRA7-based ELISA considering utilized new fragment of ge-nomic DNA is a good tool for serodiagnosis of anti- N. caninum antibodies for screening and epidemiological purposes on cattle herd and water buffaloes as well

The Effects of Seaweed Gracilaria arcuata Extract on the Stimulation of Apoptosis in Colorectal Cancer Cell Lines

Introduction: The most common problems in the medical sciences is resistance of cancer cells to anticancer drugs. Thus, finding new anti-cancer agents with minimal side effects seem necessary. In this regard, different studies on different marine algae are reported. This study aimed to investigate the effects of the algae Gracilaria arcuata extract on the stimulation of apoptosis in colorectal cancer cell lines and the effects on cell DNA fragmentation. This study was conducted as an In vitro study. Materials and Methods: The anticancer effects of aqueous and organic extracts of algae were tested by the MTT Assay, Trypan blue staining, apoptosis by flow cytometry and DNA fragmentation assay methods. Results:  The results of this study imply that the anti-cancer properties tested by MTT and Trypan blue assay, showed that methanol extract of Gracilaria arcuata with 44.45 ± 0.91 µg/mL concentration had the maximum effect on cancer cell death and the minimum effect was seen in the hexane (70.22 ± 1.22 µg/mL) and ethyl acetate extracts (70.55 ± 2.67 µg/mL), respectively (p <0.05). Results of trypan blue test confirmed the results of the MTT test, but the survival rate were different. To determine the effect of the algae methanol extract, cell apoptosis was assayed by two methods of flow cytometry and DNA fragmentation. Algae extracts of Gracilaria arcuata have LC50 in the concentration of 1010.16 µg/mL and apoptosis of 7.5 percent, which showed little apoptotic effect. Conclusion: The results show that the algae Gracilaria arcuata extract had anticancer effects against colorectal cancer cell lines, but additional studies of the chemical structure of bioactive compounds are needed

Development and Standardization of Dot - ELISA for Detection of Neospora caninum Antibodies in Cattle and Comparison with Standard Indirect ELISA and Direct Agglutination Test (DAT)

Background:Neospora caninum is a protozoan parasite from phylum apicomplexa and an important agent causing abortion in cattle which produce notable economic loss all around the world. Methods:Dot-Elisa was set up performing checker board procedure and then 178 sera of cattle examined with commercial indirect ELISA and direct agglutination test (DAT) were also evaluated by dot-ELISA afterwards.  Results:Kappa statistical analysis revealed that Dot-ELISA has good agreements with ELISA as well as the DAT and also, Mc Nemar´s analyzing showed that this procedure has acceptable ability to discriminate positive results. Relative sensitivity and specificity of Dot-ELISA were respectively 92.63% and 89.16% and 93.4% and 90.8% in comparison with ELISA and DAT.  Conclusion:Since the dot-ELISA is easy, inexpensive and not needed high experience to interpret the results, it is superior to ELISA and DAT when we aim to screen the cattle on the farm and slaughterhouses or when the laboratory equipment is not available.

The Cataleptic, Asymmetric, Analgesic, and Brain Biochemical Effects of Parkinson’s Disease Can Be Affected by Toxoplasma gondii Infection

Purpose. Parkinson’s disease (PD) is a neurodegenerative disorder with progressive motor defects. Therefore, the aim of the present investigation was to examine whether catalepsy, asymmetry, and nociceptive behaviors; the Nissl-body and neuron distribution; brain-derived neurotrophic factor (BDNF); malondialdehyde (MDA); total antioxidant capacity (TAC) levels; and the percentage of dopamine depletion of striatal neurons in the rat model of Parkinson’s disease (PD) can be affected by Toxoplasma gondii (TG) infection. Methods. Fifty rats were divided into five groups: control (intact rats), sham (rats which received an intrastriatal injection of artificial cerebrospinal fluid (ACSF)), PD control (induction of PD without TG infection), TG control (rats infected by TG without PD induction), and PD infected (third week after PD induction, infection by TG was done). PD was induced by the unilateral intrastriatal microinjection of 6-hydroxydopamine (6-OHDA) and ELISA quantified dopamine, BDNF, MDA, and TAC in the striatum tissue. Cataleptic, asymmetrical, nociceptive, and histological alterations were determined by bar test, elevated body swing test, formalin test, and Nissl-body and neuron counting in the striatal neurons. Results. The results demonstrated that PD could significantly increase the number of biased swings, descent latency time, and nociceptive behavior and decrease the distribution of Nissl-stained neurons compared to the control and sham groups. TG infection significantly improved biased swing, descent latency time, nociceptive behavior, and the Nissl-body distribution in striatal neurons in comparison to the PD control group. The striatal level of BDNF in the PD-infected and TG control groups significantly increased relative to the PD control group. The striatal MDA was significantly higher in the PD control than other groups, while striatal TAC was significantly lower in the PD control than other groups. Conclusions. The current study indicates that TG infection could improve the cataleptic, asymmetric, nociceptive and behaviors; the level of striatal dopamine release; BDNF levels; TAC; and MDA in PD rats

Neospora caninum and Toxoplasma gondii infections in one-humped camels (Camelus dromedarius) in central desert of Iran.

The protozoan parasite Neospora caninum infects carnivores as definitive and a wide range of mammals as intermediate hosts. This parasite is regarded as an important cause of abortion in cattle worldwide, causing significant economic losses. Although there is serological evidence of infection in Old World camelids, the significance of N. caninum in these animal species is still poorly understood. The aim of this study was to use molecular and histological methods to detect N. caninum in the blood and tissues of 100 slaughtered one-humped camels (Camelus dromedarius) in Iran. For this, genomic DNA was extracted from blood, brain, portal lymph node and liver of the camels, and nested-PCR assay followed by sequencing were performed. Besides, paraffin-embedded tissue sections were stained with hematoxylin and eosin (H&E) and studied microscopically. In addition, immunohistochemical staining for N. caninum was attempted on brain samples with positive PCR results. All animals were tested for antibodies against N. caninum and Toxoplasma gondii by whole tachyzoite-agglutination tests. N. caninum DNA was detected in blood, brain, and portal lymph node, but not in the liver of two (2%) camels. Histopathological examination revealed cysts resembling N. caninum in brain samples of one of these camels; however, immunohistochemical staining for N. caninum and T. gondii did not allow a morphological identification. IgG antibodies to N. caninum and T. gondii were detected in 36% and 35% of the camels, respectively. This study provides the first insight into direct detection of N. caninum in C. dromedarius in Iran. Further molecular studies on aborted fetuses, stillborn animals and cases of perinatal mortality are needed to understand the possible involvement of N. caninum in cases of reproductive failure. As the definitive hosts of N. caninum are domestic and wild canids, producers should be advised to monitor and limit exposure of their camelids to these species and their feces

Study on Theileria lestoquardiAntigens as Potential Vaccine Candidates

Theileria lestoquardi is the causative agent of malignant theileriosis of sheep and goats, causing morbidity and mortality in these animals worldwide. Western blot analysis based on T. lestoquardi schizont antigens was carried out using sera collected from Iranian sheep, which had been immunized with T. lestoquardi schizont-infected cells. The results of Western blot analysis demonstrated that schizont-immunized animals produced antibodies reacting with protein bands at 73, 42, 20, 14, and 12 kDa. Comparison of the results of the current Western blotting test with earlier studies of Theileria spp. revealed two immunogenic schizont proteins with molecular weights of 73 and 42 kDa shared between T. annulata and T. lestoquardi. Two other proteins with molecular weights of 14 and 12 kDa have not been previously found in other Theileria species. Our results suggest that the 73-kDa protein could be a potential vaccine candidate and that the 14- and 12-kDa proteins could be considered as diagnostic antigens