The effect of apricot tree gum adding to tris-base diluents on liquid semen storage and ewe pregnancy rate

Introduction: Successful artificial insemination requires successful collection,sperm evaluation and the addition of preservatives to sperm shelf life.Despite many efforts and addition of protective and antioxidant substances ,the quality of frozen-thawed sperm is low and the resulting fertility is not acceptable.For this reason,in this research,the cooling method has been used insted of freezing.The aim of this investigation was to assess the effect of apricot gum adding to tris-base diluents on liquid ram semen storage and ewe pregnancy rate. Materials and methods:This study was conducted in the khalatposhan research station of Tabriz University,EastAzarbyjanProvince,Iran.semen was collected by artificial vagina twice a week during the breeding season. A total of 25 ejaculates were collected from 5 Ghezel rams. Average age of rams was 3 years.Immediately after collection,each ejaculate was immersed into a water bath maintained at 37 ℃ prior to evaluation.The semen samples were evaluated for volume,wavemotion,spermconcentration,membraneintegrity,Ph,totalmotility,progressive motility and viability by routine examinations. The volume of ejaculate by graduated tube (shamsuddin,2000). To evaluate the wave motion,a drop of undiluted semen was placed on a pre-warmed slide 37℃ without a corerslip and examined underphase contrast microscope(100×)(Nikon,Eelpse,E200,Japan). Fresh semens have been studied in terms of volume,concentration,total motility,viability and morphology and only samples with over 2.5 billion sperm and a progressive motility of over 70% were used for dilution.Semen samples was diluted with tris(2.71g),citric acid(1g),fructose(1.4g),penicillin(100,000IU) and streptomycin(100mg) in 100 ml distilled water.Then 73 ml of this solution was mixed egg yolk(20ml). One gram of solid apricot gum was dissolved in 10ml sterile distilled water to prepare the liquid apricot tree gum. After diluting the semen samples, it was divided into four equal parts and three parts were added to 100,150 and 200 µl/ml of apricot tree gum diluting liquid,respectively,and one item(control group) no material was added.Diluted semen samples were poured into 2ml microtubes and placed in a container containing 37Ċ water in the refrigerator for 90 min to slowly reach 5Ċ.Semen samples were evaluated every three days for 36 days. In this study,12 ewes were artificial inseminated by cervical insemination and all three ewes were ineminated with one treatment.Sperm used for artificial insemination were stored for 24 hours.The artificial inseminated ewes in this experiment,were 4 years old.Data were analysed using the Glm and mixed procedures of SAS 9.2 software.
Results: In this investigation, it was observed that adding apricot gum at all levels of 100,150 and 200µl did not have a significant effect on sperm quality until the third day after cooling to tris-base diluents in all three concentrations of 100,150 and 200µl after the third day after cooling(P>0/05).but after the third day of cooling significantly improved sperm quality parameters such as viability,total motility,progressive motility,morphology and membrane health(P<0/05).but between there was not any significant difference between treatments with apricot gum(P>0/05).In this study,12 ewes were artificial inseminated by cervical insemination and all three ewes were ineminated with one treatment.Sheep inoculated with control treatment had 33.33% fertility,100 and150µl apricot gum treatment had 66.67% fertility and 200µl apricot gum treatment had 100% fertility

Mammalian sperm cells present highly specific lipidiccomposition,high content of polyunsaturated fatyacids,plasmalogenes and sphingomyelines.This unusual structure of sperm membrane is responsible for its flexibility and the functional ability of sperm cells (Sanocka and Kurpisz,2004).However,spermatozoa,s lipids are the main substrates for peroxidation ,what may provoke server functional disorder of sperm (Aitken,1989).A reason for higher,pathological lipid peroxidation of sperm membranes can be un balanced oxidative stress (Sanocka and Kurpisz,2004).The spermatozoon like all cells living under aerobic conditions,constantlv faces the oxygen paradox;oxygen is required for life,but oxidative metabolism of biological molecules can be potentially toxic because of the formation of highly reactive oxygen species(ROS)that can modify cell functions or viability (Aitken and Clarkson,1987).Reactive oxygen species leak from mitochondria in to the cytoplasm were they cause cellular damage by oxidizing a variety of biologically important molecules including DNA,proteins,lipids and carbohydrates (Przekwas,2003).Also oxidative stress by an imbalance of oxidants and antioxidants in favour of the former and is capable of inflicting injury on membrane lipids,proteins and nucleic acid (Toyokuni,1999). Endegenous antioxidant are sufficient to prevent free radicals that are produced in the normal state in the cells but an increase in free radical production leads to oxidative damage (Bunker,1992).Natural polysaccharides and their conjugates have been widely used in food and medicine fore long time.Numerous biological and pharmalogical favourable effects of natural polysaccharides have been extensively studies in a vitro and in animal models in vivo and more kinds of natural polysaccharides have been tested and even applied in therapies (Wang and Fong,2004;Liu,2016).It was demonstrated that some natural polysaccharides were effective at preventing oxidative damage in living organism this could be a potential resource of novel antioxidants (Tsiapali,2001).It is well described that the reducing capacities of polysaccharides is related to their sulfation rate molecular weight,glycosidiclinkages,hydroxyl groups but also carboxylic groups of uronic acids (Hentati,2018).It was previously reported that adirect correlation between the antioxidant activity and the reducing power of polysaccharides (El-shaurbagy GA,2014).Indeed,this antioxidant activity is explained by the presence of many free hydroxyl groups in the structureChemical analysis of apricot gum was performed in (2012) by the LIuveras-Tenoria.They found total sugars(60%),galactose(43%),mannose(4%),arabinose(44%),xylose(7%) and ramnose(1%) in apricot gum (LIuveras,2012). of polysaccharides. In addition, some papers reported that the presence of arabinos in the structure of polysaccharides could reduce the production of hydroxyl radicals by chelation of pro-oxidant ions(Lefish,2018).In a study conducted to investigate the effect of adding xanthan gum to horse sperm diluents,the researchers reported that adding xanthan gum to the diluents reduces sperm motility compared to the control group,but in other sperm quality parameters,they did not see a significant difference with the control group,which is inconsistent with the findings of our research(Gheller,2019).In another experiment to investigate the effect of substituting Gum diluents on teacling increased the storage time of semen better than the control grup and in Arabic for egg yolk in ram sperm diluents,researchers reported that substituting Gum Arabic for egg yolk increased the belief of artificial inseminated ewes which is consistent with the results of artificial insemination of sheep in our study(Zeitoun,2017). The study found that adding apricot tree gum to the tris-base treatments with apricot tree gum sperm quality is was better than in the control grup.So far,no research has been conducted to investigate the effect of adding apricot tree gum to semen diluents and this study was conducted for the first time.
Conclusion: In general,the results of this study showed that the addition of Apricot Gum to tris-base diluents due to regenerative reviews increases the quality and longevity of ram liquid semen and increases its fertility.