BACKGROUND: Storing semen at a temperature of 4 degrees Celsius reduces its motility and quality.OBJECTIVES: This study aimed to determine the effect of adding calcium compounds, including calcimaphor (CMP), calcium gluconate (CG), and calcium chloride (CaCl2), on the motility and progressive motility of rooster sperm kept at refrigerator temperature.METHODS: This research used five pieces of 45-week-old Lari breed roosters. The liquid diluent added different calcium compounds at 0.56, 0.056, and 0.0056 mM concentrations. After treatment, the diluted seminal samples were cooled at the storage temperature to avoid thermal doubt and then transferred to the refrigerator at 4 degrees Celsius. Parametric factors that were more important such as the percentage of laterality and progressive mobility, were measured visually using the lens of a 40-light microscope, survival was checked using the eosin-nigrosin staining method, acrosome health percentage by formalin citrate method, cell membrane health with hypoosmotic test, lipid peroxidation level, fertility was evaluated using perivitelline membrane sperm reaction 72 hours after storing at 4 degrees Celsius.RESULTS: Based on the results, different calcium compounds in most concentrations could significantly affect the parameters of survival, mobility, and progressive mobility (P<0.05).CONCLUSIONS: Most of the sperm-related parameters in the control group decreased after this period of storage in the refrigerator, but the addition of calcium gluconate (0.56, 0.056, and 0.0056), Calcimaphor (0.56, 0.056, and 0.0056) and calcium chloride (0.56, 0.056 and 0.0056) to the semen thinner maintained the quality indicators of rooster sperm.

BACKGROUND: Bovine Herpes Virus-1 (BHV-1) belongs to the  Alpha herpesviral family. The virus is the cause of Infectious Bovine Rhinotracheitis (IBR) and Bovine Abortion. In the initial infection, the virus proliferates excessively. Moreover, shedding the virus leads to conditions in the latent phase of the disease. Infectious Bovine Vulvovaginit (IPV ) is the genital form of the disease that represents a genital infection and transmits via pustules and mucopurulent secretions. Exposure to the virus in genital mucosa leads to IPV infection through mating or artificial insemination and the diseases that can be transmitted to healthy livestock by frozen sperm during artificial insemination.OBJECTIVES: Viral contamination of the semen is one of the routes to spread the disease among dairy cattle. Therefore, we investigated the presence of the virus in domestic and frozen imported semen consumed in industrial dairy cattle farms.METHODS: In the present study, 140 frozen straws were collected. After melting each straw, 200 µl of obtained semen was used for DNA extraction, which was done directly on the semen samples and via a Genome Extraction Kit. Subsequently, to ensure the accuracy of the extraction, the PCR technique was done using PRM-1 gene primer. Tracking the viral genome was done using the PCR technique and known primers.RESULTS: In total, one out of 140 samples was found to be virally contaminated, and IBR contamination was confirmed by repeating all the steps and determining the gene sequence.CONCLUSIONS: It is necessary to further investigate the possibility that contamination can be transmitted via frozen semen, given that even one out of 140 samples is contaminated, and the importance of the disease.

BACKGROUND: In traditional medicine in some Asian countries, including Iran, there is a belief that camphor is a suppressor of sexual activity. Not only has the validity of this hypothesis not been established, but also studies in this field are very limited.  OBJECTIVES: In this study, we investigated the effects of camphor on sperm quality in mice, and to protect sperm damage vitamin E as an antioxidant was used. METHODS: This study was conducted on 30 adult male mice (balb/c) with weight range 20-25 gr in 5 groups. First group was control (CO) and treated with normal saline,  groups  2 and 3 were sham groups treated respectively with Olive oil (OL) and the combination of olive oil and vitamin E (OL+E), and finally, two experimental groups were treated using camphor (CA) and camphor with vitamin E (CA+E). Camphor at doses of 30 mg/kg/day and vitamin E at doses of 100 mg/kg/day were prepared. All materials were administered orally (gavage). After 35 days semen were collected from tail of epididymis, and then total count, motility, viability, nuclear maturity, and DNA damage were examined. RESULTS: Results showed significant reduction in sperm total count, percentage of viability, increase in  the number of immature sperms and  no significant difference in rate of motile sperms and sperms with damaged DNA in groups that received Camphor was observed. Vitamin E as a strong antioxidant administered lightly was able to reduce the effects of  Camphor on viable and mature sperms (p<0.05). CONCLUSIONS: It can be concluded that Camphor could affect on mice sperm quality and vitamin E as an antioxidant,  was able to slightly reduce Camphor effects in sperm quality.

The Dag defect is one of the primary morphological defects in sperm correlating with reduced fertility. This defect is found in the spermatozoa of many livestock species. The aim of the study was to assess the morphometry of the heads of normal sperm and sperm with the Dag defect in the semen of Duroc breeding boars. Sperm morphology was examined in ten ejaculates each from 12 Duroc boars. In total, 3,600 morphologically normal sperm and 838 sperm with the Dag defect were evaluated. The area, perimeter, length and width of the sperm head were measured and these basic morphometric parameters were used to calculate four additional shape indices characterising the sperm head, i.e. ellipticity, elongation, roughness and regularity. Sperm with this defect had markedly smaller heads, 0.32 μm shorter and 0.19 μm narrower than the heads of sperm with normal morphological structure. The heads of sperm with the Dag defect also had a 1.1μm smaller perimeter and a 2.5 μm² smaller surface area than the heads of morphologically normal sperm. The Dag defect is found in boar sperm irrespective of the age of the individual. It affects the morphology of the sperm head.

The Dag defect is one of the primary morphological defects in sperm correlating with reduced fertility. This defect is found in the spermatozoa of many livestock species. The aim of the study was to assess the morphometry of the heads of normal sperm and sperm with the Dag defect in the semen of Duroc breeding boars.

Introduction: The effect of two smear staining methods on the dimensions and shape of sperm cells in the semen of domestic pigs was evaluated. Material and Methods: The studies were carried out on 30 ejaculates collected from 15 boars, which included five Duroc boars, five Pietrain boars, and five hybrid Duroc × Pietrain boars. Each ejaculate was next sampled to make two microscopic slides, of which one was stained with eosin-nigrosin and the other with eosin-gentian dye. In total, 600 measurements of sperm cells were made. Each sperm was measured for the following morphometric parameters: head length, head width, head area, head perimeter, tail length, and the total sperm length. Results: Sperms measured on slides stained with eosin-nigrosin showed lower dimensions as compared with those stained with the eosin-gentian dye method. Sperm stained with eosin-nigrosin had shorter and narrower heads than sperm stained with eosin-gentian dye. The method of staining, therefore, affected not only the dimensions of the sperm, but also the proportions of the dimensions defining the shape of the sperm. Conclusions: The size and shape parameters in porcine sperm may take on different values depending on the method of semen staining. Sperm cells stained with eosin-nigrosin are smaller than the sperm stained with eosin-gentian dye. The sensitivity of the sperm to the type of dye used for the fixation may be associated with genetic factors.

Introduction: The aim of this study was to evaluate the effect of lipoprotein fraction isolated from ostrich egg yolk (LPFo) on the metabolic activity of boar spermatozoa following liquid semen storage in different extenders and temperatures. Material and Methods: Boar ejaculates were extended in Androhep, Beltsville thawing solution (BTS), and Martín-Rillo and Alias (MR-A) without (control) or with the addition of LPFo and stored for three days at either 5°C or 16°C. The analysed sperm parameters included total motility (TMOT), plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), oxygen consumption, and adenosine triphosphate (ATP) production. Results: The sperm metabolic activity seemed to be higher in the LPFo-based extenders following storage for three days, irrespective of the storage temperature. Compared with the LPFo-free extenders, significantly higher (P < 0.05) sperm PMI and MMP were observed in BTS and MR-A extenders supplemented with LPFo during storage for three days at 5°C. Spermatozoa stored in the BTS-LPFo extender exhibited higher (P < 0.05) TMOT and oxygen consumption, whereas higher (P < 0.05) PMI was observed in spermatozoa stored in Androhep-LPFo and MR-A-LPFo for three days at 16°C. No significant differences (P > 0.05) in ATP content were observed between the LPFo-free and LPFo-based extenders during storage. Conclusions: Supplementation of LPFo to semen extenders had varying effects on the metabolic activity of boar spermatozoa stored at different temperatures. It can be suggested that the interactions of various components of the extenders and seminal plasma with LPFo exert beneficial effects on the sperm metabolic activity during liquid storage of boar semen.

Introduction: The effect of two smear staining methods on the dimensions and shape of sperm cells in the semen of domestic pigs was evaluated. Material and Methods: The studies were carried out on 30 ejaculates collected from 15 boars, which included five Duroc boars, five Pietrain boars, and five hybrid Duroc × Pietrain boars. Each ejaculate was next sampled to make two microscopic slides, of which one was stained with eosin-nigrosin and the other with eosin-gentian dye. In total, 600 measurements of sperm cells were made. Each sperm was measured for the following morphometric parameters: head length, head width, head area, head perimeter, tail length, and the total sperm length. Results: Sperms measured on slides stained with eosin-nigrosin showed lower dimensions as compared with those stained with the eosin-gentian dye method. Sperm stained with eosin-nigrosin had shorter and narrower heads than sperm stained with eosin-gentian dye. The method of staining, therefore, affected not only the dimensions of the sperm, but also the proportions of the dimensions defining the shape of the sperm. Conclusions: The size and shape parameters in porcine sperm may take on different values depending on the method of semen staining. Sperm cells stained with eosin-nigrosin are smaller than the sperm stained with eosin-gentian dye. The sensitivity of the sperm to the type of dye used for the fixation may be associated with genetic factors.

Introduction: The aim of this study was to evaluate the effect of lipoprotein fraction isolated from ostrich egg yolk (LPFo) on the metabolic activity of boar spermatozoa following liquid semen storage in different extenders and temperatures.

This study was performed to investigate the reproductive performanceand spontaneous malformations of female Sprague Dawley (SD) rats obtained from the Animal Resource Unit, Institute for Medical Research for their use in reproductive toxicity assessment. Onehundred and thirty-five virgin female rats with a body weight ranging between 190 g and 210 g were randomly caged overnight with 60 fertile male rats in 1:1 basis. Vaginal smear was performed the next morning and the day of sperm positive was considered as estational day (GD) 0. On GD21 caesarean hysterectomy was performed to examine the outcome of each pregnancy. The female fertility index (sperm positive) was 90.5% and the pregnancy index was 100%. Thepercentage of pre and post-implantation loss were 1.96 and 7.48 respectively with the number of early resorption out numbered the late resorption (0.56 ± 1.0 and 0.36 ± 1.5 respectively). Out of1,234 foetuses examined, only 5 foetuses (0.41%) presented with gross congenital malformation. The female SD rats obtained from the Institute demonstrated high fertility and pregnancy indexes with lowincidence of resorption and malformation, therefore suitable to use for reproductive toxicity assessment.