BACKGROUND: Semen cryopreservation process with increased reactive oxygen species levels (ROS) plays a decisive role on the sperm cellular organelles (cell membrane, mitochondria and DNA). OBJECTIVES: The aim of this study was to determine the effects of adding different levels of superoxide dismutase (SOD) and reduced glutathione (GSH) antioxidants before freezing on motility, total progressive motility, viability and positive response to hypo-osmotic swelling test (HOST) parameters after Bull semen was frozen. METHODS:  In this study, four bulls aged 4-5 years were used and 20 times the semen collection (5 ejaculates per bull) was performed. SOD (100 and 150 IU/ml) and GSH (5 and 7.5 mM) antioxidants were added to Tris - egg yolk extender. Diluted semen samples with or without additives were manually filled with straw and the free end of the straws were sealed by sealing machines; then, semen were frozen by semi-automatic freezing machine, using liquid nitrogen. After thawing the samples, the total and progressive motility, sperm viability and positive response to the HOST parameters were determined at zero and 2 h of incubation. RESULTS: Adding SOD and GSH antioxidant levels did not reveal significant differences between treatments in any of the semen parameters in the initial hour of incubation after thawing. Two hours incubation of post-thawed semen samples containing SOD (100 IU/ml) significantly improved total motility, viability and membrane integrity of the sperm (p<0.05). Also, in samples containing SOD (150 IU/ml), sperm membrane integrity was maintained (p<0.05). In the semen samples with antioxidant GSH (5 mM), total motility and membrane integrity of the sperm are considerably improved after thawing (p<0.05). CONCLUSIONS: This study showed that using SOD and GSH antioxidants as freezing semen extender additives can  improve post-thawed bull semen quality.

Background: Haemoproteus is a parasitic protozoa that over 120 species of it has been reported from wild aquatic birds, sparrows and other birds orders. So far, no study has been performed to determine the species of this protozoa in the north of Iran.Objectives: The aim of this study was to investigate the molecular and structural properties of Heamoproteus protozoa in the blood of infected pigeons in Mazandaran province.Methods: In the present study molecular investigation of Haemoproteus infection was carried out in domestic pigeons of Mazandaran province. For this purpose, samples obtained randomly from 150 pigeons in different regions of Mazandaran. At first, blood samples were stained with Gimsa stain and examined for presence of Haemoproteus gametocytes. Then, positive samples were used for PCR by Cytochrome b genes.. Results: Obtained results after morphological survey showed that 17 samples were positive indicating infection rate of 11.33%. Molecular investigation and analysis of PCR products showed that all of the samples belonged to Haemoproteus columbae species.Conclusion: So, being precisely familiar with this kind of protozoan and its species can limit many mistakes and be helpful in differential diagnosis of different species. This study has revealed that the most common species of Haemoproteus in Mazanadran province is Haemoproteus columbae.

AbstractBACKGROUND: Beluga (Huso huso) is one of the sturgeons and currently is cultured artificially in Iran with different goals. The cardiovascular system is one of the important systems of the body, and heart is the most important organ in this system. So far, few studies have been done on the heart of sturgeons.OBJECTIVES: This study was done to determine the exact structure of beluga different parts of the heart septum using echocardiographic and histologic techniques. METHODS: Six immature male belugas (2.5 years old) were investigated in this study. For echocardiography, probe was placed on the ventral surface of body, between pectoral fins. At first, macroscopic morphologic studies of different parts of heart were done at the topographic place of them. For histological studies, usual methods of fixation and hematoxylin and eosin (H&amp;E) coloring were used. Prepared slides were studied by light microscope.RESULTS: All parts of the heart including the sinus venosus, atrium, ventricle, conus arteriosus and pericardium were clearly visible in different probe moves. The pericardium were covered surround heart`s structures having homogenous echo pattern. Sinus venosus had a very thin septum without any expansion and contraction. Atrium had a completed muscular septum with expansion and contraction. Ventricle muscular septum had two layers: an external compact layer and an internal spongy layer. Conus arteriosus was composed of obvious muscular septum and connective tissue. Histological studies showed three layers in beluga heart including epicardium, myocardium and endocardium. In overall, the histological findings were matched with the results of echocardiography. CONCLUSIONS: This study is the first investigation on echocardiographic feature of different parts of beluga heart. These results were coordinated with histological findings. Normal information and findings of this study can be used for investigation of various anatomic or pathologic changes in beluga heart in different research projects.

BACHGROUND: Epilepsy is a chronic neurological disorder. Despite discovery of effective antiepileptic drugs (AEDs), more than 30% of patients are still resistant to AEDs. OBJECTIVES: Evaluation the effect of acute administration of Docosahexaenoic acid (DHA) in mice resistant to antiepileptic drugs in 6Hz model of epilepsy. METHODS: at first we evaluted intracerebroventricularly (i.c.v) injection and oral consumption of DHA alone and intrapritoneal (i.p.) injection of drugs in separate groups. In test groups LTG 25mg/kg or PHT 25mg/kg were injected i.p. 105 min after injection of PHT, 45 min after injection of LTG, DHA (1mM) was injected i.c.v. In control groups drugs solvent or DHA solvent was injected DHA. 15 min after injection of DHA or DHA solvent, in all groups 6 Hz stimulation was exerted and occurrence of limbic seizures was registered. In oral test groups PHT 25mg/kg or LTG 25mg/kg were injected i.p. 60 min after injection of PHT and simultaneous injection of LTG, DHA (0.1 ml) was gavaged. 60 minutes after injection of DHA 6 Hz stimulation was exerted. RESULTS: Acute administration of DHA alone via i.c.v injection or oral gavage had no protective effect on inhibiting seizures. Administration of DHA with LTG or PHT also couldn’t inhibit drug resistance. 6-Hz seizures when administered chronically. However, chronically administered DHA inhibited limbic seizures resistant to LTG and PHT. CONCLUSIONS: Acute administration of DHA couldn’t inhibits resistance to LTG and phenytoin in 6-Hz model of epilepsy. Also consumption of single dose of DHA with anticonvulsant drugs don’t have any effect on prevention of drug resistance in epileptic patients.

BACKGROUND: PRP (Platelet-Rich Plasma) is a biological product that has shown significant therapeutic results compared to synthetic drugs and some traditional methods. The advantages of this method include: PRP, compared to other biological materials, is easily prepared in clinic and the safety is high as well. OBJECTIVES: We aimed to investigate the evaluation of histomorphometric properties after injection of plasma. METHODS: Thirty rats were divided randomly into three groups: control group (normal saline), PRP and PPP (Platelet-Poor Plasma) groups. In order to prepare PRP and PPP, blood samples (1cc) were collected through cardiac puncture and were centrifuged. The upper layer consisted of PPP and lower layer consisted of PRP. The 2% calcium gluconate solution (100 µl) was added to platelet-rich plasma (200 µl) layer to activate platelet. After seven days, the dorsal skin samples were collected. They are cut into 5 to 6 μm thickness sections using paraffin embedding method and were stained by hematoxylin, eosin and Masson’s trichrome. The epidermis, dermis and hypodermis layers of skin, the epithelial thickness of hair root sheath, maximum depth of hair follicles, the thickness of the whole skin, the number of sebaceous glands and hair follicles in the specified scale, finally the number of fibroblast cells and the percent distribution of connective tissue fibers were investigated in histometric structure of the skin and skin appendages. RESULTS: The results of this study indicated that systematic and controlled PRP may have beneficial effects on the reconstruction and rehabilitation of the skin and its appendages. CONCLUSIONS: The present study showed that growth factors in plasma especially PRP, could be stimulated connective tissue fibroblast of dermal layer in the skin.

BACKGROUND: The most common metal used in the body is iron and since it may produce ROS, it could potentially be a dangerous substance. OBJECTIVES: Regarding the side effects of nanomaterials such as nano iron particles and possibility of environmental contamination including air pollution with nanoscale iron particle, this study was designed to compare the effects of conventional iron oxide with iron oxide nanoparticles, on certain indexes of ovarian tissue in a rat experimental model. METHODS: This animal model was carried out in 5 groups of female rats, including control, iron oxide (15 mg) and iron oxide nanoparticles (5, 15 and 30 mg). The drugs were intraperitoneally injected daily for 16 days. On the seventeenth day the rats were euthanized by chloroform. Ovarian tissue was removed, and histological changes and iron accumulation were assessed by special staining and light microscopy. RESULTS: According to our findings, folliculogenesis was decreased in all groups receiving iron. The number of corpus luteum in the groups receiving different doses of nanoparticle was reduced and the number of atretic follicles was significantly increased in all groups compared to the control group. CONCLUSIONS: In conclusion, probably iron nanoparticles with impaired cellular oxidative pathways, reduces the number of follicles and corpus luteum and increase atretic follicles by producing oxygen free radicals and destructing microfilaments. This can cause a negative effect on the fertility of female rats.

BACKGROUND: Diazinon is an organophosphate pesticide which is widely used in paddy fields located in northern parts of Iran, though it is prohibited to be used in many countries. OBJECTIVES: The purpose of the present study was to investigate sublethal effects of diazinon on blood parameters of common bream Abramis brama. METHODS: In sublethal toxicity test, fish were exposed to diazinon concentrations of 0.04, 0.36, 0.73 and 1.46 mg/l for 14 days and their hematological parameters including WBCs, white cell differential count, RBCs, Hct, Hb, MCV, MCH and MCHC were studied.  RESULTS: The results of sublethal toxicity indicated that by increase of toxicant concentration a significant decrease was appeared in WBCs, RBCs, Hct, Hb and MCH both after 7 and 14 days (p<0.05). After 7 days a significant decrease was observed in lymphocytes count which accounted for about %81 of the control group. The neutrophils count was also face the same reduction so that after 14 days of diazinon exposure, the neutrophils counted decreased to about %85 of control group. CONCLUSIONS: Regarding the toxicity of diazinon on various hematological parameters and the diazinon concentration in Iranian waters and considering the fish habitat in the waters which are connected to the Caspian Sea it suggests that the pesticide diazinon affect the fish survival.

BACKGROUND: This study was conducted to evaluate the effect of melatonin on semen quality and quantity changes and increase of reproductive efficiency in Lori-Bakhtiari ram in the non-reproductive season. OBJECTIVES: The purpose of this study was to investigate the effect of melatonin on testicular size and semen qualitative and quantitative changes in the non-breeding season is Lori-Bakhtiari rams. METHODS: This study was designed to evaluate the effects of melatonin on the quantity and quality of semen and testicular diameter in 8 Lori- Bakhtiari rams (4 control rams and 4 treatment rams) for 10 weeks during spring 2013. Each ram in the treatment group was received three tablets of 18 mg implantable melatonin subcutaneously at the base of ear after 5 weeks from the start of the trial. RESULTS: Semen parameters and testicular diameter measurements were evaluated on a weekly basis. No significant differences were observed between 2 groups before melatonin implanting. A significant difference was observed after melatonin implanting between 2 groups in the mean of testicular diameter (treatment: 33.5±0.25 and control: 30.67±0.07 cm), in semen volume (treatment: 1.34±0.03 and control: 0.87± 0.04 ml) and in sperm concentration (treatment: 1.76±0.05 and control: 1.37±0.04 109/ml) (p<0.05). However, other parameters including average of lives sperm percentage, total sperm motility, sperm progressive movement and percentage of abnormal sperm were not significant (p>0.05). CONCLUSIONS: The results of this experiment showed that the use of melatonin in the non-reproductive season resulted in increase of testicular size, semen volume and sperm concentration in Lori-Bakhtiari ram. However, the rest of the semen parameters have not been affected.

BACKGROUND: Yersinia ruckeri is the etiological agent of enteric red mouth (ERM) or yersinioisis disease, one of the important bacterial diseases in the cultured salmonids. OBJECTIVES: The purpose of present study was detection of gyrA gene (quinolone resistance) in the Y. ruckeri bacterium. METHODS: In this study fish were evaluated in average size 8-12 cm from six rainbow trout farms in Chahar Mahal va Bakhtiyari province (Iran). In each farm 10 fish (totally 60) suspected to yersinioisis were randomly selected; sampling was done from lower part of intestine and cultured on Trpticase Soy Agar (TSA). The mediums were transferred to incubator and kept at 22 °C for 48 hours. Pure colonies which are grown on the mediums were tested by catalase, oxidase and gram staining, then those of gram-negative, catalase positive and  oxidase negative were diagnosed, and cultured on Waltman- Shots medium (as specific medium for Y. ruckeri). These mediums were incubated at 22 °C for 48 h. Colonies that were grown were tested by PCR method for Y.ruckeri detection. Then, in the identified strains of Y.ruckeri gyrA gene were detected by PCR test. RESULTS: The results of bacteriological, biochemical and molecular tests showed that three cases out of total isolates were identified as Y. ruckeri. In all isolates of Y. ruckeri, gyrA gene was identified by molecular test. CONCLUSIONS: Identification of quinolone resistance gene in Y. ruckeri isolates can be the reason of low efficacy of these classes of antibiotics in the aquaculture. ِTherefore, the policy of treatment should be changed specially in enteric red mouth disease.

BACKGROUND: In recent years, use of herbs as dietary irritant growth factors and replacing it with artificial materials in aquaculture is discussed. Rosemary (Rosmarinus officinalis) oil is one of herbal medicines which its antioxidant and antibacterial properties have been proved in humans and animals. So it can be an alternative option in order to achieve artificial growth stimulants properties. OBJECTIVES: The purpose of this study was to investigate the effect of different levels of rosemary oil as a medicinal herb on growth parameters and gut morphology of beluga juveniles (Huso huso). METHODS: Six groups (three replicate/group) of 126 fish with initial mean body weight of 130.94 ± 5.28g were fed with diets containing 0 (control), 0/01, 0/1, 1 and 2 % of rosemary oil and a group of oxytetracyclin (30mg/kg) for 8 week. At the end of the trial, the growth parameters (final weight, specific growth rate and feed conversion ratio) were calculated. Then, beluga intestinal biopsy was performed and the mean villi length, width and surface were measured in the treatments. RESULTS: Significant increase were observed in villi length (2/25±0/11mm), width (0/52±0/01mm) and surface (1/17±0/07mm) in fish fed with diets containing 1ml/kg rosemary oil (p<0/05). There were no significant differences in growth parameters between treatments. CONCLUSIONS: According to the significant increase of beluga intestinal absorption area and slight improvement of growth parameters in fish fed with diets containing rosemary extract compared to the control group, it seems that rosemary oil can act as a growth stimulant. So it can be considered as an alternative to artificial growth stimulants in aquaculture industry.