Testing Usability of Butylated Hydroxytoluene in Conservation of Goat Semen

Abstract

The objective of this study was to investigate whether butylated hydroxytoluene (BHT) could be used as a suitable supporter or alternative of egg yolk during preservation of goat spermatozoa. Three in vitro experiments and a fertility test were conducted to evaluate the effect of BHT on viability of chilled-stored semen as well as motility and kidding rate of frozen-thawed spermatozoa. In the first two experiments, ejaculates (n = 30/experiment) were collected from 10 bucks, split, diluted with egg yolk-based and egg yolk-free extenders supplemented with or without 0.3, 0.6, 2, 5 and 8 mM BHT and stored at 5 degrees C for 168 h. In the third experiment, 30 ejaculates were collected from the above-mentioned bucks, split and diluted with egg yolk-free extenders supplemented with or without 0.3, 0.6 and 0.9 mM BHT and egg yolk-based extenders supplemented with or without 5 mM BHT. Diluted semen was cooled to 5 degrees C over a period of 4 h, frozen and thawed in the form of 0.3-ml pellets. In the fertility test, 75 ejaculates were collected from two proven fertile bucks, split, diluted with egg yolk-free extenders containing 0.6 mm BHT and egg yolk-based extenders supplemented with or without 5 mM BHT, frozen and thawed as described above. An insemination volume of 0.6 ml containing 120-140 x 10(6) progressively motile spermatozoa was used for a single cervical insemination of cloprostenol-synchronized does (n = 230). The results showed that addition of 5 mM BHT to egg yolk-deficient (2.5%) extenders significantly improved viability of chilled-stored semen together with motility (48.5%) and fertility (62.5%) of frozen-thawed spermatozoa. Replacement of egg yolk in semen extenders by 0.6 mm BHT could sustain not only viability of chilled-stored semen but also post-thaw motility (47.5%) and fertility (53.75%) of frozen-thawed spermatozoa. In conclusion, supplementation of semen diluents with BHT can ameliorate preservability of goat sperm.

Optimal inclusion level of butylated hydroxytoluene in semen extender improves the quality of post-thawed canine sperm

“…However, owing to the paucity of literature regarding the use of antioxidants in canine species, the results need to be discussed in comparison to their use in other animals/ species. Similar improvements in semen quality by the use of BHT have been reported for turkey tom (Donoghue and Donoghue, 1997), cattle bull (Chaterjee and Gagnon, 2001;Shoae and Zamiri, 2008), ram (Watson and Anderson, 1983), goat (Khalifa et al, 2008), and boar (Roca et al, 2004) spermatozoa. The protective effect of antioxidant (BHT) is attributed to two mechanisms: firstly, the incorporation of the compound in the sperm membranes, hence, making them more fluidic and preventing them from the damage (Ijaz et al, 2009) and, secondly, declining the damaging potential of lipid peroxyl radicals by conversion into hydroperoxides (Thomassen et al, 2006).…”

“…The damage inflicted on the sperm membrane during cryopreservation renders the membrane more permeable to the BHT metabolites which, in turn, has a harmful effect (Shoae and Zamiri, 2008;Sahashi et al, 2011). Similarly, BHT was found to be toxic for cattle bull (Shoae and Zamiri, 2008), goat (Khalifa et al, 2008), human (Aitken and Clarkson, 1988) and boar (Roca et al, 2004) spermatozoa. This toxicity is due to counteracting the ROS-induced oxidative stress by higher antioxidant levels, hence hampering those functions of spermatozoa which are associated with ROS.…”

“…Butylated hydroxytoluene (BHT) is one such example being tested for its cryoprotective potentiality. BHT has been used successfully for preservation of liquid semen in turkey tom (Donoghue and Donoghue, 1997) and to minimize cryoinjury in ram (Watson and Anderson, 1983), boar (Roca et al, 2004), cattle bull (Shoae and Zamiri, 2008), and goat spermatozoa (Khalifa et al, 2008). However, there are scanty reports (Neagu et al, 2010;Sahashi et al, 2011) regarding its cryoprotective potentiality and optimal inclusion level in canine semen.…”

Effect of Butylated Hydroxytoluene on Dog Sperm Longevity in Chilling Storage and Cryopreservation

“…Supplementation of Uppsala extender and INRA-96 extender with 1 mM BHT did not improve post-thaw motility of dog sperm [16], which is 897 EFFECT OF BHT ON DOG SPERM STORAGE consistent with our findings. However, BHT has been demonstrated to maintain post-thaw sperm characteristics in many species [13,14,18,24]. Two mechanisms for protecting sperm by BHT have been suggested [10]: 1) BHT is incorporated into the sperm membrane, resulting in an increase of membrane fluidity, and 2) BHT prevents the activity of lipid peroxyl radicals by converting them to hydroperoxides.…”

“…Supplementation of semen extenders with antioxidant compounds could overcome this problem. Butylated hydroxytoluene (BHT), a synthetic phenolic antioxidant, is not only commonly used as a food additive [26], but is also widely used for low-temperature preservation of mammalian [2, 3,11,13,14,17,18,24,25] and avian sperm [6]. It was reported that BHT suppressed sperm mitochondrial metabolism and protected the cell membranes from cold shock by decreasing oxygen toxicity in the bull [12].…”