ABSTRACT:The objective of this study was to assess the sperm membrane integrity in extended boar semen during storage time using specific spectrum laboratory methods. Boar semen was diluted with the long-term liquid commercial extenders Androhep (A), Androstar (AS), Androstar plus (AS + ), LD and M III and was stored up to 96 h. The sperm membrane integrity was evaluated by motility, viable spermatozoa, short hypoosmotic swelling test (sHOST) and by the activity of the enzyme aspartate aminotransferase (AST). Negative changes in the quality of sperm membrane in relation to storage time were observed after 48 h for sHOST, after 72 h for viable spermatozoa and after 72 h for motility. The percentage of viable spermatozoa was decreased by 0.27% each hour. A statistically significant difference between extenders A and LD was observed in sHOST after 72 h and 96 h storage (P < 0.05). The AST activity did not show any statistically significant differences in extenders and in storage time. In overall assessment Androhep was the best of the tested extenders, followed by AS, AS + , M III and LD in terms of motility, viable spermatozoa and sHOST. The correlations among laboratory methods were highly significant (P < 0.001). In conclusion, the results documented that the sperm membrane integrity was statistically significantly affected by extenders and storage time (P < 0.001). Boar semen quality was the best in extender A. sHOST is a very sensitive and relatively simple method for the assessment of sperm membrane integrity in diluted semen.
Some shape abnormalities of spermatozoa (AS) can be a result of pathological processes that affect testicles and the epididymis tract, others can be caused genetically. Some may be caused by unsuitable rearing conditions. Some authors of the papers published in this field consider the level of AS occurrence to be an important marker for the semen quality besides the sperm motility (Leidl et al., 1971;Lyczynski and Pawlak, 1974;Blom and Andersen, 1975;Gamčík et al., 1976;Blom, 1977;Stemmler et al., 1982;Wekerle, 1982;Falkenberg et al., 1984;Yoshida and Kojima, 1989;Waberski et al., 1990Waberski et al., , 1994Itoh and Toyama, 1995;Itoh et al., 1996;Věžník et al., 2000;Louda et al., 2001;Corcuera et al., 2002;Gadea, 2002). According to Waberski et al. (1990) two criteria are sufficient for the selection of boars for insemination or ejaculate: sperm motility and percentage of AS, especially when the semen is preserved and used for the insemination of sows after a longer time, i.e. three to five days. Larsson et al. (1988) considered the motility and morphology of spermatozoa to be the most sensitive indicators of the heat stress of boars. Malmgren (1989) The aim of the paper was to find out the level of changes in the sperm quality in two groups of boars in the insemination (A and B) with diametrically different contents of morphologically abnormal spermatozoa (AS) with an interval of 25 weeks between semen collection I and collection II. In the group A there were 22 boars with the AS content up to 10%, in the group B 16 of boars with the AS content above 40% in collection I. Both groups were comparable concerning the parameters of the performance test results and in quantitative parameters of the semen from collection I. They differed significantly in the AS content (P < 0.01) and in the age (P < 0.05). In collection II in both groups the semen volume increased significantly (A -P < 0.01; B -P < 0.05), in the group A the number of spermatozoa per ejaculate and per day also increased (P < 0.01). In the group B there was an insignificant clear decrease in the sperm concentration (P > 0.05). In comparison with the group B the group A can be characterized as a group with significantly higher dynamics in the sperm production per ejaculate. An opposite trend was noted in the total AS content. In the group A there was a significant increase (P < 0.001) and in the group B a significant decrease (P < 0.001) in collection II. In the group A there was a deterioration of the AS content in 7 boars (31.8%), in the group B an improvement in 7 boars (43.7%). Particular monitored AS forms are given. A significant difference in the total AS content between both groups was in favour of the group A (P < 0.001). While no boars from the group A exceeded the limit in the AS content for the applicability of semen for insemination (in the Czech Republic 25%), in the group B they remained above this limit without applicability possibility. The detected variations and prevailing stability in the AS occurrence in boars kept in the same condi...
The objective of this study was to analyze the influence of the year-season effect on semen production parameters in the fertile AI boars. The evaluation was especially focused on the morphologically abnormal spermatozoa (MAS) incidence. It was microscopically evaluated after making fresh semen smears and staining on microscopic slides. MAS incidence 19.46 % was lower at first half-year than at second half-year 25.00 % (P<0.01). Spermatozoa with distal protoplasmic droplet were furthest participated in total MAS incidence. Its rate was the highest at fourth quarter in comparison with annual period (P<0.001). We found that season has the negative effect on sperm morphology and significantly affects boar sperm quality and subsequently AI dose quality.Keywords: boar; morphologically abnormal spermatozoa; seasonal effect ABSTRAKTCílem této práce bylo zjistit vliv ročního období na kvalitu spermatu plemenných kanců se zvláštním zaměřením na výskyt a strukturu morfologických změn spermií. Abnormality spermií byly posuzovány mikroskopicky, prostřednictvím obarveného nátěru nativního spermatu na podložním sklíčku. Sezónní výskyt morfologicky abnormálních spermií lze komentovat nižším výskytem v prvém pololetí 19,46 % a vyšším výskytem ve druhém pololetí roku 25,00 % (P<0,01). Spermie s distální protoplazmatickou kapkou se podílely v největší míře na celkovém výskytu morfologicky abnormálních spermií, přičemž ve srovnání se zbývající částí roku byl jejich podíl nejvyšší ve čtvrtém čtvrtletí roku (P<0,001). Bylo shledáno, že vliv ročního období má negativní efekt na morfologii spermií a výrazně ovlivňuje kvalitu spermatu kanců a následně i kvalitu inseminační dávky. Jan LIPENSKÝ, Alena LUSTYKOVÁ, Josef ČEŘOVSKÝ DETAILED ABSTRACTCílem předložené studie bylo analyzovat vliv ročního období (sezóny roku) na dynamiku kvalitativních parametrů spermatu kanců používaných celoročně na ISK k produkci inseminačních dávek se zvláštním zaměřením na výskyt a strukturu morfologických změn spermií. U souboru kanců, chovaných v relativně optimálních podmínkách výživy a ošetřování, byly sledovány změny kvality spermatu v závislosti na sezóně roku, resp. podle čtvrtletí (I. leden -březen, II. duben -červen, III. červenec -září, IV. říjen -prosinec). Hodnocení výskytu morfologicky abnormálních spermií (MAS) probíhalo v období let 2006 až 2008, přičemž zpracováno a mikroskopicky vyhodnoceno bylo celkem 623 vzorků od 68 kanců různé plemenné příslušnosti. Posuzovány byly morfologicky abnormální změny na hlavičce spermií, bičíku, akrozómu, dále výskyt proximálních protoplazmatických kapek (PPK) a distálních protoplazmatických kapek (DPK) na bičíku a výše nezařazené jako vady ostatní. Tabulka 1 srovnává procento MAS dle sledovaných let a jejich čtvrtletí. Sezónní výskyt MAS lze komentovat nižším výskytem v prvém pololetí 19,46 % a vyšším výskytem ve druhém pololetí 25,00 % (P<0,01). Podrobná analýza podílu jednotlivých skupin vad spermií z celkového počtu MAS zjištěných v jednotlivých čtvrtletích za celé sledované období je prezentována v...
AbStrAct:The objective of this experiment was to test a hypothesis that L-carnitine supplemented diet would improve semen characteristics in 6 adult crossbred AI boars (Hampshire × Pietrain). The control and the tested diet were identical except the tested period (8 weeks) when the diet was supplemented with 2 g of l-carnitine per boar per day. The semen was collected regularly weekly by a gloved-hand technique. Semen volume, sperm motility and concentration, proportion of alive sperm cells and aspartate aminotransferase (AspAT) activity of semen were determined immediately after the semen collection once every two weeks. Sperm survival rate, morphologically abnormal spermatozoa, seminal plasma mineral components and free amino acid concentration, l-carnitine concentration in semen plasma and in sperm cells were determined after the sample storage (-20°C) at a later time. The differences ascertained between the average values of semen characteristics in the control vs. tested period did not prove a true and unambiguous positive effect on boar semen parameters by dietary supplementation of l-carnitine as our data show in our study: volume (239.11 vs. 250.50 ml; P 0.518), sperm concentration (301.67 vs. 350.83 × 10 3 /mm 3 ; P 0.309), sperm progressive motility (66.94 vs. 70.00%; P 0.409), morphologically abnormal spermatozoa (29.00 vs. 27.46%; P 0.802), daily sperm cells output (9.86 vs. 11.71 × 10 9 ; P 0.206), proportion of alive sperm cells (72.56 vs. 74.13%; P 0.484), survival spermatozoa ability maintenance (43.29 vs. 38.68%; P < 0.01), mineral components in the seminal plasma (Na-, K-, Ca-, Mg-, Zn-; P from 0.138 to 0.968), AspAT activity (in semen plasma -132.50 vs. 128.31 mU/10 9 spermatozoa; P 0.830, in sperm cells -147.37 vs. 119.01 mU/10 9 spermatozoa; P 0.146), semen plasma amino acid concentration -a significant positive effect of L-carnitine in lysine only (0.79 vs.1.17 µmol/100 ml; P < 0.01), l-carnitine concentration (in semen plasma 255.40 vs. 259.97 mg/l; P 0.884, in sperm cells -1 110.68 vs. 883.58 mg/l; P < 0.01). In conclusion, the studied indicators of semen quality were not significantly enhanced by dietary supplementation of l-carnitine in adult AI boars.
ABSTRACT:The objective of this study was to provide some data on concentrations of seminal plasma free amino acids (FAAs) in order to compare these values with different total contents of morphologically abnormal spermatozoa (MAS) in the semen of 37 experimental adult boars. The boars were kept in the same housing, feeding and breeding conditions. Immediately after collection, semen gel free volume, sperm concentration and total MAS were determined microscopically. The boars were divided into two groups (A -n = 24, B -n = 13) according to the significantly different total MAS content ( -x: A = 9.46 ± 4.95, B = 46.00 ± 14.54%, P < 0.01). Deproteinized seminal plasma samples (supernatant) were analysed for concentrations of 13 FAAs: Tau, Asp, Thr, Ser, Glu, Gly, Ala, Val, Met, Ile, Leu, Tyr and Lys by liquid chromatography (AAA 339 M analyser). A highly significant correlation was determined between total MAS and total FAA content in all experimental boars (n = 37, r = -0.60, P < 0.01). Glutamic acid was a predominant FAA in seminal plasma in both groups of boars ( -x: A = 28.49 vs. B = 17.60µM/100 ml) but its concentration was significantly higher in group A (P < 0.01). The proportion (%) of glutamic acid concentration in the total content of FAAs in both groups of boars was nearly equal (A = 38.1 vs. B = 38.9%, P > 0.05) as well as the proportion index of the individual FAAs calculated from glutamic acid (= 100%). The differences in the particular FAAs recorded between group A and group B were statistically significant for 11 out of the 13 FAAs observed (P < 0.05 and P < 0.01) in favour of group A as well as in the total calculated content of FAAs per boar ( -x: 74.70 vs. 45.23µM/100 ml, P < 0.01). A significantly negative relationship between the MAS content and the concentration of seminal plasma FAAs (r = -0.60, P < 0.01) is the main result of this study with regard to the markers for potential boar semen fertility estimation.
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