Repression of common bull sperm flora and in vitro impairment of sperm motility with Pseudomonas aeruginosa introduced by contaminated lubricant

Semen collected from clinically healthy bulls at an artificial insemination centre was examined for bacterial diversity. While bacteria that are normally present in the common flora of bovine semen were absent, such as Mycoplasma sp., Proteus sp. and Corynebacterium sp., all semen samples contained an unusually high number of Pseudomonas aeruginosa strains. Analysis via pulsed field gel electrophoresis demonstrated that one particular P. aeruginosa strain, present in a sealed bottle of lubricant, was widespread in bull semen. This strain was shown to secrete substances that inhibited both the growth of bacteria constituting the normal bull sperm flora and the motility of spermatozoa in vitro. This study demonstrated that commercially available lubricants might contain bacteria that can spread amongst breeding bulls and affect the quality of semen. Bacteriological controls and species' identification are necessary at several production levels, including lubricants and extenders, to ensure high semen quality and avoid the spread of pathogens.

Assessment of BoviPureTM for the In Vitro Production of Bovine Embryos

The objective of this study was to examine the potential utility of a commercially available sperm separation and purification product for the in vitro production of bovine embryos. Bovine oocytes were purchased from a commercial supplier, and matured oocytes were randomly allocated to one of two treatments. Oocytes were co-incubated with frozen-thawed semen washed twice with BoviPureTM (BoviPure group) or with modified Brackett-Oliphant medium (control group). After a 6-hour insemination period, oocytes were cultured in vitro for 8 days. Cleavage rate of embryos was determined 48 hours post-insemination, and blastocyst formation rate was assessed on day 8 of culture. The experiment was replicated three times, and data were analyzed using chi-square analysis. Washing of sperm in BoviPureTM had no effect (P>.05) on either cleavage rate (77.2%) or blastocyst development (21.6%) when compared with controls (71.9% and 17.1%, respectively). These results indicate that, under conditions of our study, the washing of sperm with BoviPureTM did not significantly enhance the ability to produce bovine embryos in vitro.

Importance of sperm transition proteins demonstrated in mice carrying Tnp1- and Tnp2-null alleles

Chromatin condensation within the nucleus of developing spermatids involves replacement of histones by transition proteins, which are in turn replaced by protamines. The importance of transition proteins in the complex process of spermiogenesis has, to date, been only speculative. This study sought to investigate the extent to which transition proteins are essential or have redundant functions by characterizing sperm produced in mice expressing all combinations of Tnp-null alleles. Results from breeding trials of 8 weeks duration revealed that, on average, wildtype males produced about 14 offspring whereas TP2 and TP1 single-knockout males produced about 8 and 1 offspring, respectively, demonstrating their subfertility. Genotypes with less than two Tnp wildtype alleles, as well as double-knockout mutants, were completely infertile. Sperm from males with impaired fertility had poor progressive motility, heterogeneous chromatin condensation, incompletely processed protamine 2 and head and tail abnormalities. Generally, as the number of Tnp-null alleles increased so did the severity of abnormalities. However, specific morphological abnormalities were associated with the absence of an individual TP. Studies which sought to identify possible root causes for abnormalities in thiol-rich sperm structures revealed no differences in thiol content or sulfhydryl oxidation status within the nucleus but nuclei and tails from single-knockout mutants were severely disrupted following thiol reduction. Binding of fluorescent dyes to DNA was normal in sperm recovered from caput but abnormal in cauda epididymal sperm from TP1 knockouts and infertile double mutants. Injection of cauda epididymal sperm from double knockouts into oocytes produced very few offspring; however, after injection with testicular sperm, the efficiency was no different from wildtype. These results suggest DNA structural alterations or degradation during epididymal transport of sperm resulting in a diminished capacity of the paternal DNA of these sperm to produce offspring. The overall importance of transition proteins for normal chromatin condensation and production of fertile sperm has been demonstrated. Furthermore, identification of specific morphological abnormalities associated with the absence of an individual transition protein provides new evidence that the proteins are not completely redundant and each fulfills some unique function. ^

Seawater carbonate chemistry and sperm swimming speed of the polychaete Galeolaria caespitosa in lab experiment

The rapidity of ocean acidification intensifies selection pressure for resilient phenotypes, particularly during sensitive early life stages. The scope for selection is greater in species with greater within-species variation in responses to changing environments, thus enhancing the potential for adaptation. We investigated among-male variation in sperm swimming responses (percent motility and swimming speeds) of the serpulid polychaete Galeolaria caespitosa to near- (delta pH 0.3) and far-future ocean acidification (delta pH 0.5). Responses of sperm swimming to acidification varied significantly among males and were overall negative. Robust sperm swimming behavior under near-future ocean acidification in some males may ameliorate climate change impacts, if traits associated with robustness are heritable, and thereby enhance the potential for adaptation to far-future conditions. Reduced sperm swimming in the majority of male G. caespitosa may decrease their fertilization success in a high CO2 future ocean. Resultant changes in offspring production could affect recruitment success and population fitness downstream.

Viabilility of Rams Epididymal Sperm after Preservation in Low Temperature (5oc)

The purpose of this research was to evaluate the viability of ram epididymal sperm collected from fresh caudal epididymis (H-0) or after storage in low temperature (5oC, in refrigerator) for one (H-1), two (H-2), and three (H-3) days.  Collected sperm were diluted in modified Tris extender and they were preserved in refrigerator up to four days.  The viability of diluted sperm was evaluated daily base on motility and sperm live.  Results indicated that mean sperm concentration after sperm diluted with 0.05 ml Tris extender of caudal epididymis was 2745 million/ml. Sperm motility and percentage of live for H-0 (71.25% and 82.83%) and H-1 (70.00% and 79.17%) were significantly higher (P<0.05) than H-2 (61.25% and 69.83%) and H-3 (51.67% and 66.17%).  Percentages of sperm motility and live of diluted sperm and preserved in refrigerator for H-0 were significantly higher (P<0.05) than H-1, H-2, and H-3.  These results showed that epididymal sperm collected from caudal epididymis up to three days of preservation (without further storage of the diluted sperm) could be used for artificial insemination or in vitro fertilization programs.  Diluted sperm of H-0 and H-1 could be preserved in refrigerator for two days and H-2 for one day. (Animal Production 6(1): 30-36 (2004) Key Words: Epididymal Sperm, Viability, Rams

The effect of Physiological NaCl Dilution levels and Storage Duration to Sperm Motility and Abnormality in Muscovy Duck

The aim of this research are to know the effects of physiological NaCl dilution levels and storage duration on sperm motility and abnormality of  Muscovy duck. Materials which used in this research were semen that collected of 1  - 1,5 years of four Muscovy duck. Semen were treated by physiological NaCl dilution levels nol , three, six and nine and storage duration ( t0 = 0 minute t1 =45minutes and t2=90 minute). Replication was three tapping periode . The method of this research was laboratory experimental with Randomized Complete Blok Design (RBD) in Split Plot Design (SPD)  pattern ,as main plot is Physiological NaCl dilation levels and as sub plot is storage duration. Result of the experiment showed that physiological NaCl dilution levels was very significantly influences on sperm motility (p<0,01) and significantly influences and sperm abnormality (p<0,05) . The storage duration was very significantly influences and sperm motility and abnormality (p<0,01) .Orthogonal duration polynomial assayed showed that the effect of physiological NaCl dilution levels to sperm motility  had a linier regression line with the a equation as Y =69.94 - 2.65 X,  r=0,49 R2=0.24 with Y presented to sperm motility and X represented to physiological NaCl dilution levels  sperm and effect of storage duration to sperm motility had a linier regression line with the equation as Y =74.24-0.35 X, r =0.73 R2=0.53 with Y represented to sperm motility and X represented to storage duration . The effect of physiological NaCl dilution levels to sperm abnormality had a linier regression line with the equation as Y =5.61+0.34X, r=0.52 R2=0.27 with Y represented to sperm to abnormality and X represented to regression line with the equation as Y =6.74+0.01X, r=0.24R2=0.060 with Y represented to sperm abnormality and X represented to storage duration. It is can be concluded that a higher physiological NaCl dilation levels and storage duration caused sperm to motility decreased and sperm abnormality increased. (Animal Production 3(2): 45-52 (2001)Key Word: sperm Muscovy duck.

Quality Of Pe Buck Sperm Stored In Different Packaging

AI for cattle has been develop in Indonesia in contrast, AI for small ruminants (sheep and goat) are less developed. Its due to the lack of facilities, processing and packaging during storage. This research aims to compare the effectivity and efficiency of two different packaging technique (test tube and straw). Test tube and 0.25 ml mini straw were tested . Semen was obtained from adult PE buck (3 yr) after collection by using  artificial vagina. The fresh semen was then evaluated and diluted 5 folds with 2.9 percent Na-citrate. Diluted semen then packed in test tube and mini straw, and stored in refrigerator (100C) for 7 days. Observations were done everyday on sperm motility, abnormality and percentage of  live sperm. Observation were made at 370C. Observation on fresh ejaculate showed that semen has 6x 109/ml concentration, 90 percent motility, 8 percent abnormality and  95 percent  live sperm. Five folds dilution reduced sperm concentration to 1.2 x 109/ml, but did not change sperm motility, abnormality and percent of live sperm. Sperm was then packed according  to the treatments. Storage in both packaging did significantly reduce  (P<0.01) sperm motility and percent live sperm, but not for sperm abnormality. All sperm stored in test tube were classified death at day 5 storage. However, sperm in the straw were remain live eventhough the percentages was low. It can be concluded that straw has a better result than test tube. Eventhough sperm motility was extremely low, it remains valuable for cervical insemination. (Animal Production 1(1) : 24-29 (1999).  Key Words : Sperm, PE Buck, Straw, Test Tube

The Effect of Level Testosteron e Addition in Diluents and Level Dilution on Speed Movement and Abnormality of Kedu Chicken Sperm

The research entitled “The Effect of Level Testosterone Addition in Diluents and Level Dilution on Speed Movement and Abnormality of Kedu Chicken Sperm” was conducted in Laboratory of Physiology and Reproduction, Faculty of Animal Science, UNSOED, started at August 15th to September 15th , 2001. The aims of  this research were to obtain influences of level testosterone and dilution on speed movement and secondary abnormality kedu chicken sperm and obtain interaction between the treatments. The tapped sperm from nine kedu chickens were used in this research. This Experiment was performed 4 x 3 Factorials with Randomized  Completely Block Design (RCBD) as the basic design. The treatment combinations were level testosterone 0, 300, 600 and 900 μg (t0, t1, t2 and t3) and level of dilution 4, 6 and 8 time (in a row p1, p2 and p3). The tapping period was replicated four times as a group (replicated) with two days interval. The result of this research showed that the interaction between level of testosterone addition and level of dilution gave a non-significant effect to speed movement sperm but significant to abnormality of kedu chicken sperm. The group gave a significant influence (P<0.05) to speed movement sperm and non significant to sperm abnormality. The interaction of level testosterone addition and level dilution of kedu chicken semen (T x P) has a quadratic regression to sperm abnormality with regression comparison is Y = 24.418 – 0.014 X +1.187E – 05 X², with peak point is (543.76: 20.23) of correlation coefficient (r) 0.55 and determination coefficient (R²) as 30.34%. The addition of 600 μg testosterone level with 6 level dilutions was the best to defend sperm abnormality. (Animal Production 4(2): 60-70 (2002) Key Words : Spermatozoa, Kedu Chicken, Testosterone, Dilution, Speed Movement and Abnormality