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Search for Bovine Herpes Virus I in Iranian Frozen Semen
2022
Arabkhalegh, Fateme | Mirshokraei, Pezhman | Seifi, Hesamoddin
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.
Show more [+] Less [-]Evaluation of sperm quality in mice exposed to camphor and protective role of Vitamin E
2016
adibmoradi, masood | kalantari hesari, ali | morovvati, Hassan | asadi, farzad | Moradi, Hamid Reza
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.
Show more [+] Less [-]Supplementation of Lake Extender with Calcium Compounds to Maintain the Quality Parameters of Rooster Sperm for 72 Hours
2022
Farzam, Parand | Farzinpour, Amjad | Vaziry, Asaad | Naderi, Sina
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.
Show more [+] Less [-]Assessment of the morphometry of heads of normal sperm and sperm with the Dag defect in the semen of Duroc boars
2021
Banaszewska Dorota | Andraszek Katarzyna
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.
Show more [+] Less [-]Use of alternative extender added of fructose aiming the cryopreservation of boar semen
2018
Ludymila Furtado Cantanhêde | Eduardo Nunes de Freitas | Tatyane Bandeira Barros | Daianny Barboza Guimarães | Aline Viana Dias | Ricardo Toniolli
This study aimed to analyze skimmed milk powder (SMP) and fructose in a new cooling curve to freeze boar semen. A total of 49 semen samples from seven boars were cryopreserved using the new curve with addition of glucose and fructose to the refrigerating diluents: Beltsville Thawing Solution (BTS + G; BTS + F) and Skimmed milk powder (SMP + G; SMP + F), totaling four experimental groups for analysis. To finish the curve, aliquots of semen were packaged in 0.5 ml straws and kept in liquid nitrogen. During the cooling curve, SMP mean spermatic vigor and motility were greater than the BTS (p < 0.05). After thawing, a decrease of spermatic force and motility in both extenders was observed, where the BTS presented spermatic vigor (2.1 ± 0.55) and motility (38 ± 21.8), presenting better results (p < 0.05). There was no statistical difference between sugars added to the BTS and SMP in spermatic force and motility (p > 0.05), although the use of fructose allowed an equalization of motility between the SMP and BTS (p > 0.05). Functionality of membrane was better preserved with the addition of fructose, in both extenders. The rate of sperm viability was significantly higher in extender containing glucose and SMP (71.8 ± 12.5). The percentage of intact acrosome was higher on the treatment containing glucose, independent of the extender (BTS + G: 81.8 ± 7.2, SMP + G: 81.4 ± 14.2). To conclude, the results suggest that the BTS is still the best option to cryopreserve and fructose could be used in boar semen cryopreservation in new cooling curve.
Show more [+] Less [-]Assessment of the morphometry of heads of normal sperm and sperm with the Dag defect in the semen of Duroc boars
2021
Banaszewska, Dorota | Andraszek, Katarzyna
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.
Show more [+] Less [-]Application of two staining methods for sperm morphometric evaluation in domestic pigs
2017
Kondracki, Stanisław | Wysokińska, Anna | Kania, Magdalena | Górski, Krzysztof
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.
Show more [+] Less [-]Metabolic activity of boar semen stored in different extenders supplemented with ostrich egg yolk lipoproteins
2017
Dziekońska, Anna | Kinder, Marek | Fraser, Leyland | Strzeżek, Jerzy | Kordan, Władysław
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.
Show more [+] Less [-]Effects of tris (hydroxymethyl) aminomethane and egg yolk on the cryopreservation of buck semen
2022
Md. Mostofa Kamal | Md. Emtiaj Alam | Md. Akhtarul Islam | Md. Royhan Gofur | Aurangazeb Kabir
Objectives: This study was designed to examine the effects of various concentrations of tris (hydroxymethyl) aminomethane (tris) and egg yolk on the quality of cryopreserved buck sperm. Materials and Methods: The collected semen samples were pooled, washed, and diluted into five different freezing extender groups, viz., extender I (tris 0% + egg yolk 0%), extender II (tris 1.41% + egg yolk 4%), extender III (tris 2.41% + egg yolk 8%), extender IV (tris 3.41% + egg yolk 16%), and extender V (tris 4.41% + egg yolk 24%). The sperm parameter of the five groups of extenders was evaluated after equilibration and cryopreservation. Results: The results showed that extenders II–V provided significantly higher semen progressive motility and total motility percentages than extender I after equilibration (p < 0.05). The higher percentages of semen progressive motility, total motility, viability, and plasma membrane integ¬rity (by both HOST under light microscopy and stain after HOST under light microscopy) were found in the sperm cryopreserved with extender IV than extender I, extender II, and extender III groups after thawing (p < 0.05). In addition, semen progressive motility, total motility, and viability were not further increased, or plasma membrane integrity (by both HOST tests) was decreased by the addition of tris and egg yolk (extender V) after cryopreservation (p < 0.05). Conclusion: In conclusion, our result indicates that the following washing, the supplementation of tris (3.41% + egg yolk 16%) on the freezing extender are suitable for improving the semen quality of buck after freezing and thawing. [J Adv Vet Anim Res 2022; 9(4.000): 676-683]
Show more [+] Less [-]Single and mixed interaction of experimental Trypanosoma brucei brucei and Trypanosoma evansi on the semen collection reaction time and spermatozoa morphology of Yankasa rams
2016
Yunusa A. Wada | Sonnie J. Oniye | Peter I. Rekwot | Oluyinka O. Okubanjo
Objective: This study was conducted to evaluate the pathological effects of experimental trypanosomosis on the semen collection reaction time and spermatozoa morphology of Yankasa rams. Materials and Methods: Twelve apparently healthy Yankasa rams aging 24-30 months and weighing 22-25 Kg were randomly selected and were distributed into four (4) groups. Groups I and II were challenged with experimental Trypanosoma brucei brucei (Federe strain) and T. evansi (Sokoto strain) respectively, while group III was challenged with both T. brucei brucei and T. evansi parasites. Group IV was left as uninfected control. Each infected ram received 2 mL of the infected blood containing 2x106 trypomastigotes via the jugular vein. The animals were examined for clinical observations, reaction time for semen collection and abnormalities in the morphology of the spermatozoa.Results: Infection of rams with trypanosomes showed scrotal edema, scrotal atrophy, loss of libido, increased semen collection reaction time, and significant increase of spermatozoa morphological abnormalities in all the infected rams. The rams especially in groups I and III were all deemed unfit for breeding by the end of the 98 days post infection, while the uninfected rams remained as healthy and had normal values of sperm morphology throughout the study period. Conclusion: Single or mixed interaction with T. brucei brucei or T. evansi is capable of causing infertility and reproductive failure in Yankasa rams. [J Adv Vet Anim Res 2016; 3(4.000): 360-367]
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