Estimation of endogenous levels of osteopontin,total antioxidant capacity and malondialdehyde in seminal plasma: Application for fertility assessment in buffalo (Bubalus bubalis) bulls

This study was attempted to identify subfertile bulls by quantifying the endogenous levels of osteopontin (OPN), total antioxidant capacity (TAC) and malondialdehyde (MDA) in seminal plasma of buffalo bulls. On the basis of conception rate, buffalo bulls were classified into two groups: high‐fertile (conception rate >50%) and subfertile bulls (conception rate <40%). A total of 100 ejaculates (10 ejaculates from each bull) were collected through artificial vagina method. The concentration of OPN, TAC and catalase (CAT) of high‐fertile bulls was found to be higher (p < .05) than that of subfertile bulls. Further, MDA level in seminal plasma was found to be lower (p < .05) in high‐fertile bulls compared with subfertile bulls. The fertility status had no effect on the superoxide dismutase (SOD) concentration in seminal plasma of both the groups. The levels of OPN (r = .678, p = 0.013) and TAC (r = .648, p = .042) were found to be positively correlated with bull fertility and the level of MDA (r = ?.718, p = .019) was found to be negatively correlated with bull fertility. However, the fertility of bulls was not found to be significantly correlated with SOD, CAT and sperm motility. In conclusion, seminal OPN, TAC and MDA tended to be more realistic in identification of subfertile bulls from breeding herds.     

Die Bedeutung von Hoden- und Nebenhodenanom alien,insbesondere von Hodendrehungen bei Höhenfleckvieh- und Braunviehstieren für die Fruchtbarkeit1

Contents 1. 206 bulls, 13–24 months old, were tested from the A. I. centre (Spotted Mountain Cattle and German Brown Cattle). The measurements of the testes, the semen quality on the basis of 10 ejaculates of each bull, and the fertility quotient by means of the 60 to 90-Days-Non-Return results of a three months utilization (about 800 to 1000 first inseminations) were determined beside a general clinical investigation of the sexual organs. 2. From the tested bulls 12% showed a rotation of the testes about the longitudinal axis, 4,35% comparatively small testes, 2,9% an asymmetry of the testes, and 3,4% an underdevelopment of the cauda epididymidis. 3. A decrease of the semen quality and of the fertility quotient was determined in bulls with small testes, with asymmetrical testes and with an underdevelopment of the cauda epididymidis. The rotation of the testes had no influence on the fertility and quality of the sperm. 4. The results of the clinical tests, especially the size of the gonades must be given a greater consideration in the selection of stud bulls to ensure an optimal fertility.     

Effect of pre-freeze addition of cysteine hydrochloride and reduced glutathione in semen of crossbred Jersey bulls on sperm parameters and conception rates

This study was aimed to assess the effect of pre-freeze addition of cysteine hydrochloride and glutathione (GSH) on post-thaw sperm functional parameters and field fertility. The experimental bulls (n = 6) aged 4-6 years were used for this study. A total of 36 ejaculates (six ejaculates per bull) were collected and divided into three groups, group I (control), group II (5 mm cysteine hydrochloride) and group III (5 mm GSH). The extended semen samples with and without additives were filled in mini straw using automatic filling and sealing machine and cryopreserved in liquid nitrogen. Post-freeze seminal traits were recorded after thawing at 37°C for 30 s. The curvilinear velocity (VCL) and amplitude of lateral head displacement values were significantly (p < 0.05) higher in GSH than the cysteine and control groups. Post-thaw plasmalemma integrity was significantly (p < 0.01) higher in GSH group when compared to cysteine and control groups. The loss of acrosomal integrity was significantly (p < 0.01) lower in GSH than in cysteine and control groups. Addition of GSH non-significantly (p = 0.1) improved mitochondrial membrane potential (MMP) (per cent) than control group. This study indicates that the addition of cysteine and GSH suggestive (p = 0.08) of reducing lipid peroxide levels. The conception rate (%) in glutathione group was significantly (p < 0.05) higher than that in cysteine (68 vs 58) and control (49) groups. The post-thaw sperm progressive forward motility (r = 0.4) had moderate, but no significant correlation with conception rate. However, post-thaw straight-line velocity (r = 0.7), loss of acrosomal integrity (r = -0.8) and MMP (r = 0.9) had significant (p < 0.05) correlation with field fertility. This study indicates that the use of glutathione as semen additive may be used for improving post-thaw semen quality and overall augmentation of pregnancy in cows.     

Effect of dihydrostreptomycin or oxytetracycline on reproductive capacity of bulls

Young beef bulls (n = 27) were used in a trial to study the effect of dihydrostreptomycin sulfate (DHS) or oxytetracycline (OTC) hydrochloride on spermatogenesis, epididymal sperm maturation, and freezability of sperm. Nine of the bulls were given a 22 mg/kg dose of DHS twice, 12 hours between doses. Nine other bulls were treated with OTC--1 dose of 26.4 mg/kg of body weight, and then 6 more doses each of 17.6 mg/kg, ca 12 hours between doses. The remaining 9 bulls were nontreated controls. The treatment regimens with the 2 antibiotics were without effect on spermatogenesis. These treatments also were without effect on seminal pH, ejaculate volume, percentage of motile spermatozoa, rate of spermatozoal motility, or concentration of spermatozoa in ejaculates harvested on day 3 or 7 of the study (day 0 = 1st day of treatment). There was a treatment-by-day effect on spermatozoal concentration; the number of sperm per milliliter was markedly increased on day 3 for OTC-treated bulls. The increased spermatozoal concentration in the OTC-treated group was associated with an influence of the antibiotic on ejaculation. All bulls given this antibiotic ejaculated without palpable penile engorgement or erection on day 3. On day 7 the rate of spermatozoal motility was increased in the 2 treatment groups compared with the rate in the control bulls. Also on day 7, the percentage of motile spermatozoa was greater in the OTC-treated bulls than in the control or DHS-treated bulls.(ABSTRACT TRUNCATED AT 250 WORDS)     

饲养管理方式对甘南牦牛繁殖力及杂交后代影响的研究

[目的]为了研究加强种公牛和母牦牛的饲养管理对母牦牛的繁殖效果和杂交后代生长发育的影响.[方法]本试验用大通牦牛和其冻精杂交甘南牦牛,通过对试验组的种公牛、母牦牛和杂交后代进行补饲、延长放牧时间和提前断奶等措施,研究了203头母牦牛的繁殖效果和其杂交后代的生长发育情况.[结果]表明,自然交配试验组发情率、受胎率、繁殖成活率分别比对照组提高9.19%、11.03%、14.10%,流产率和犊牛死亡率降低3.3%、3.87%;人工授精组发情率、受胎率、繁殖成活率分别提高6.74%、5.48%、10.52%,流产率和犊牛死亡率降低4.07%、4.62%.加强母牦牛的饲养管理,所产犊牛的初生重比传统饲养管理条件下母牦牛所产的犊牛初生重公母分别增加2.92 kg和2.07 kg,提高19.80%和14.22%.6月龄体重分别增加12.82 kg和12.23 kg,提高17.45%和16.88%.[结论] 用大通牦牛改良甘南牦牛,采用人工授精和自然交配都有良好的效果,加强种公牛、母牦牛饲养管理是提高牛群繁殖力的有效方法,对后代生长发育影响明显.     

The Effect of Season on Semen Characteristics and Freezability in Bos indicus and Bos taurus Bulls in the Southeastern Region of Brazil

The objectives of this study were to evaluate the effects of season in southeast of Brazil comparing genotypes on semen characteristics, freezability and peripheral plasma concentrations of testosterone. Ejaculates of five Bos indicus bulls and six Bos taurus bulls were evaluated over a period of 27 months, which was divided into winter (July, August, September), spring (October, November, December), summer (January, February, March) and autumn (April, May, June). Semen was evaluated according to standard procedures for ejaculate volume, sperm concentration, gross-motility, progressive motility and sperm morphology. After preparing and freezing the ejaculates according to commercial procedures, the straws were stored in liquid N₂ until post-thaw evaluation. Ejaculate volume, sperm concentration, gross-motility, progressive sperm motility, vigor and morphological sperm defects were significantly influenced by season and genotype (p < 0.05). Heat tolerance was better in B. indicus bulls than in B. taurus bulls characterized by lower values of sperm abnormalities throughout the observation period. The highest values were recorded for abnormal heads followed by cytoplasmatic droplets in B. taurus bulls. The proportion of ejaculates which were eliminated before freezing for reasons of bad quality was lower in the B. indicus bulls. Temporal changes in peripheral plasma testosterone concentrations were higher in B. indicus bulls than in B. taurus bulls not revealing seasonal influences. The results of this study show clear genotype differences regarding semen quality. Freezability of B. taurus semen varies considerably throughout the year, leading to a high proportion of eliminated ejaculates. Collecting semen from B. taurus bulls during the summer in an artificial insemination centre may not be profitable.     

Relationship between serving capacity of beef bulls as predicted by the yard test and their fertility during paddock mating

Six experiments were conducted to determine the relationship between the serving capacity of bulls as predicted by a 40-min yard test and their fertility during paddock mating, measured by the conception rate at first oestrus and the pregnancy rate at the end of 10 weeks of mating. Twenty bulls varying in serving capacity from 1 to 11 were mated to 40 heifers each. As serving capacity of the bulls increased from 1 to 7, conception rate increased from 18 to 70%. Average conception rates achieved by 4 bulls with low serving capacity (25.3%), 8 bulls with medium serving capacity (61.4%) and 7 bulls with high serving capacity (72.3%) were all significantly different from one another. Bulls of low serving capacity (1 or 2 services) impregnated a significantly lower proportion of their heifers (40.3%) than bulls with medium (91.2%) or high (95.3%) serving capacity. It was concluded that bulls of serving capacity 1 or 2 (in 40 min) should be considered unsound for breeding. An explanation for the results and their implication in beef production is discussed.     

Isolation of Chlamydia psittaci from bull ejaculate

During the repeated serological examination (RVK) in five breeding bulls the positive levels of antibodies to Chlamydia psittaci in titre 1 : 128 were found. In the isolation experiments the pelleted ejaculates deposited in liquid nitrogen were used. The isolation of Chlamydia psittaci on yolk sacs of chicken embryos was positive in two breeding bulls. The isolated strains are labelled GN-33 and OK-107. The serological examination of blood samples was in all five breeding bulls negative on brucellosis (BAB), infectious bovine rhinotracheitis and coxiellosis and positive on PI-3. Bacteriological examination of spermatic fluid proved only sporadic contamination with moulds and saprophytic bacteria.     

Changes in sperm ultrastructure of bulls with decreased sperm activity and freezability]

A decrease of sperm freezability occurred at the K. breeding station, and this situation lasted longer than a year. Out of the 2550 ejaculates taken from 42 breeding bulls within 12 months, 685, i.e. 26.7%, were unfit for use immediately after sperm collection, mostly owing to a low activity of spermatozoa and pathological forms of their motility, and another 469 ejaculates, i. e. 18.3%, were unfit for use after sperm freezing; on the whole, 1154 (i. e. 45.2%) ejaculates had to be excluded. It was revealed by the vital-lethal primuline test that the spermatozoa died quickly after collection. The findings obtained during an electron-microscopic examination of the spermatozoa at the beginning of the process included visible changes in the ultrastructure of the flagellum, particularly its middle piece (deformed shape, incomplete set of axial filaments, vacuolization of the flagellum, abnormal arrangement of the mitochondrial spiral), numerous abnormities of the external cytoplasmic membrane and invagination, vacuolization, and abnormal density of nucleoplasm. The primary changes on the flagella and in the nucleus give evidence that the testicular tissues were altered. The etiological factors behind these processes are believed to include a reduction in the resistance of bulls due to long-lasting consumption of feeds contaminated with the fungus Aspergillus fumigatus, insufficient movement and bad zoo-hygienic practices, all this combined with the secondary action of the infectious germs of Mycoplasma bovigenitalium, which were revealed by cultivation tests in 50% of the ejaculates of the bulls; a positive antibody titre was demonstrated in all bulls.     

Bestimmung der osmotischen Resistenz von Eberspermien und deren Beziehungen zur Konservierungsfähigkeit von Samenproben

Contents Determination of the osmotic resistance of boar spermatozoa and the relations to sperm freezing and long storage The osmotic behaviour of fresh boar spermatozoa (482 ejaculates from 34 A.I. boars) was examined in a standard medium but with different osmotic pressures ranging from 150 mosm/kg till 500 mosm/kg. The percentage of spermatozoa with normal acrosomal ridge (NAR) was determined after incubation of 0.2 ml fresh semen in 3 ml of the osmotic test solutions at 39° C for 15 and 120 min. In the non-isotonic media % NAR decreased and could be quantified depending to incubation time and to the different osmolalities; NAR decreased strongest at 150 or 500 mosm/kg and after 120 min incubation time. Differences in % NAR were large between ejaculates and between boars but low within boars and statistically significant correlations had been found to % NAR and motility in froren/thawed semen samples or stored samples. Still higher correlations were found by calculation of an osmotic resistance test-value (ORT-value) which is defined as follows: ORT = 1/2 % NAR in isotonic medium/(after 15 min) +% NAR in 150 or 500 mosm/kg/(after 15 or 120 min) ORT-values can be calculated for hypotonic (150 mosm/kg) as well as for hypertonic (500 mosm/kg) test solutions and for 15 and 120 minutes incubation time. The hypotonic ORT-value for 120 min. is the most accurate; the values varied from 34 till 83 between ejaculates and from 44 till 76 between boars with a low variability within boars. Grouping of boars by their ORT-values in “good”, “moderate” and “poor” corresponded to a similar grouping of these boars by their semen quality (% NAR and motility) in preserved semen samples. Also a grouping of single ejaculates by their ORT-values corresponded to a similar qualitative classification of these ejaculates. ORT seems to become a good and practicable assessment test for the selection of boars suitable dor deep freezing or long storage of their spermatozoa. On the other hand preservation techniques too can be examined by ORT. But more investigations with this test must be done especially its relation to the fertilization capacity of semen samples with different ORT-values.     

Vergleichende Betrachtung über den Einsatz tines mobilen Phantoms (holländisches Modell, Hersteller: J.P. Tasche u. Zoon, Fleringen, Oldenzaalsweg 103 / Holland) und von "Standbullen" bei der Samengewinnung in einer Rinderbesamungsstation.

Inhalt Es wird über Erfahrungen bei der Samengewinnung beim Bullen mit Hilfe eines mobilen Phantoms über einen Zeitraum von 4 Jahren berichtet. Von 84 eingesetzten Bullen nahmen 59 das Phantom regelmäßig an. In dem genannten Zeitraum wurden 2 563 Ejakulate am Phantom und 1 064 Ejakulate unter Verwendung eines Bullen als Sprungpartner gewonnen. Der Vergleich einiger Ejakulatsmerkmale (Volumen, Dichte) ließ keine wesentlichen Unterschiede zwischen den beiden Bullengruppen erkennen. Auch bezüglich der NR-Rate bestanden keine erheblichen Differenzen. Die Vorteile des Phantoms beruhen auf seiner Mobilität, der veränderbaren Höheneinstellung and in hygienischer Hinsicht. Weiterhin ist das Verletzungsrisiko, das beim Einsatz von “Standbullen” besteht, ausgeschaltet. Contents (Comparative studies on the use of a mobile phantom (Dutch model, manufacturer: J.P. Tasche & Son, Fleringen, Oldenzaalsweg 103, Holland) and “standing bulls” for semen collection in an artificial insemination centre for cattle). The report records experience of semen collection in bulls by means of a mobile phantom over a period of 4 years. Of 84 bulls, 59 used the phantom regularly. During the period studied 2 563 ejaculates were collected with the phantom and 1 064 with the use of another bull as partner. A comparison of the ejaculates collected by the two methods showed no significant differences in volume or density. There were also no differences in the NR-Ratio. The advantages of the phantom are its mobility, adjustable height and greater hygiene. In addition the risk of injury, which exists with standing bulls, is eliminated.     

Variability in plasma membrane integrity, mitochondrial membrane potential and acrosomal status before and after cryopreservation of bull sperm

In the present study variability of bull sperm concerning percentages of sperm with intact plasma membranes (PMI), high mitochondrial membrane potential (HMMP) and positive acrosomal status (PAS) before and after cryopreservation (vKK; nKK) between bulls and between ejaculates within bulls was examined. Studies were performed on 4 semen samples each of 20 Deutsche Fleckvieh bulls. VKK-Values were 76.5% +/- 9.6% (PMI) 68.3% +/- 8.9% (HMMP) and 9.8 +/- 5.1% (PAS) and nKK-values were 38.1 +/- 14.0% (PMI), 38.2 +/- 14.0% (HMMP) and 30.9 +/- 12.1% (PAS). After freezing, variabilities in sperm parameter values between bulls (nKK: PMI: 49.8%, HMMP 52.1% and PAS: 56.6%) were nearly quite as high or higher than variabilities between ejaculates (nKK: 50.2%, 47.9% and 43.4%). VKK-values of PMI, HMMP and PAS were only fairly to moderately related (0.36 < r < 0.53; P < 0.05) to nKK-values. The results show that PMI, HMMP and PAS did not only vary between bulls, but also between ejaculates within bulls. As there are no high relationships in these sperm parameters between times before and after cryopreservation, each ejaculate should be examined after cryopreservation in order to receive a reliable information about the quality of cryopreserved sperm.     

Seminal shedding of bluetongue virus in experimentally infected mature bulls

Sixteen bulls ranging in age from 2 to 11 years were experimentally inoculated with bluetongue virus to investigate the frequency and duration of seminal shedding of the virus. All bulls developed typical viremia, lasting 21 to 58 days, and they seroconverted 2 to 3 weeks after inoculation. Virus isolation was attempted from a total of 232 ejaculates, 163 (70%) of which were collected during the period of viremia. Bluetongue virus was not isolated from any of the ejaculates collected from 11 of the 16 bulls. Virus was isolated from 9 of 52 ejaculates collected from the other 5 bulls during the period of viremia. In no instance was virus isolated from semen without concurrent isolation from blood.     

Evaluation of two seminal collection regimens for mature Holstein bulls

Twenty mature Holstein bulls (3 to 10 yr old) were used to test the effect of two semen collection regimens on spermatozoal output, post-thaw percentage spermatozoal motility, and time needed to make the collections/week. For both regimens, six ejaculates/wk were collected using either three ejaculates/d, 2 d/wk, or two ejaculates/d, 3 d/wk. A three-period switchback experimental design was used. Each collection period for which measurements were taken was 3 wk and was preceded by a 2 wk period of acclimation. The total number of spermatozoa harvested per week was not significantly different (P greater than .05): 33.2 X 10(9) when the bulls were collected two ejaculates 3 d/wk, compared with 33.9 X 10(9) three ejaculates 2 d/wk. Post-thaw progressive spermatozoal motility was 50.3 and 52.1% (P greater than .05), respectively. The average time per week to collect each bull was 73.6 and 83.7 min (P less than .05), respectively.     

Vergleich einiger Methoden zur Untersuchung von Bullensperma***

Inhalt Im Rahmen eines Versuches mit experimenteller Hitzebelastung wurden insgesamt 555 Ejakulate von 29 Jungbullen mit verschiedenen spermatologischen Methoden untersucht. Zwischen fast allen Untersuchungsergebnissen wurden hochsignifikante Korrelationen festgestellt. Für die zusammenfassende Beurteilung verschiedener Bullengruppen konnten bereits folgende Parameter als Grundlage dienen: Prozentsatz vorwärtsbeweglicher Samenzellen, Farbstoffabsorptionstest und Anteil der Spermien mit einem normalen apikalen Rand (NAR) der Kopfkappe. Weniger aussagefähig waren u. a. pH-Wert und intrazelluläre Glutamat-Oxalacetat Transaminase(GOT)-Aktivität. Contents Comparison of some methods of investigation of bull semen In a thermal loading experiment 555 ejaculates from 29 young bulls were investigated by means of different spermatological methods. Highly significant correlations could be ascertained between most of the results. The following parameter could be used for evaluating the different groups of bulls. Percentage of progressive motility, supravital stainings and number of spermatozoa with a normal apical ridge of the acrosome (NAR). Certain methods, for example, pH-Value and intracellular activity of glutamicoxalacetic-transaminase (GOT) proved less useful.     

GnRH和LHR基因多态性和牛精液品质性状的相关分析

本试验采用PCR—RFLP方法分析了GnRH基因外显子2和LHR基因内含子9在144头中国荷斯坦牛和79头河南地方肉牛品种中的多态性,利用最小二乘法分析多态位点不同基因型与精液品质性状的关系。研究结果表明,2～3岁荷斯坦牛的鲜精活力显著高于4岁以上的牛,而畸形率显著低于7岁以上的牛。对2～4岁荷斯坦牛不同精液品质性状的简单相关分析表明,畸形率与顶体完整率和冻精活力呈显著的负相关（相关系数r分别为-0．736和-0．500）。不同基因型与精液品质性状的关联分析结果表明,144头中国荷斯坦牛所研究位点不同基因型对精液品质性状没有显著影响。而河南地方肉牛GnRH基因外显子2的A883G位点GG基因型的精子密度显著低于AA和AG基因型．LHR基因G51656T位点的TT基因型精子密度显著高于GT基因型,未检测到GG基因型。并且发现随着年龄的增长,种公牛的精液品质逐渐变差。GnRH和LHR基因可作为影u向肉牛精液品质性状的候选基因。     

Replication requirements and number of ejaculates needed for assessing treatment effects on sperm output and seminal characteristics of electroejaculated Holstein bulls

Two ejaculates were harvested by electroejaculation on each of 3 d per week for 14 wk from 14, 12- to 24-mo-old Holstein bulls. Ejaculates during the first 2 wk served to stabilize sperm output. Data for the remaining ejaculates were used in a components of variance approach to determine the number of bulls per treatment and ejaculates per bull that would be needed to provide adequate sensitivity and precision for assessing treatment effects on seminal characteristics. The latter included the number of sperm (total and motile), volume and sperm concentrations of the sperm-rich fraction, and the percentage of progressively motile sperm in first and second ejaculates or per day. Replication requirements declined as the number of ejaculates per bull was increased to about 15 to 25 but declined minimally thereafter. The number of bulls needed per treatment varied in relation to the size of the treatment response to be detected but was much greater than the replication used in typical experiments. Replication for assessing sperm output in first ejaculates or per day was approximately one-half as great for experiments in which pretreatment information was used to adjust post-treatment data. Tables should be useful as a guide for designing efficient, cost-effective studies of known sensitivity and precision.     

Elektronenmikroskopische Untersuchungen zum Vergleich von gut and mangelhaft einfrierbaren Bullenejakulaten

Inhalt Die Spermatozoen von Bullen, deren Ejakulate gut, mangelhafl and nicht einfrierbar waren, wurden vor and nach dem Einfrieren elektronenmikroskopisch untersucht. Als Ursache der mangelha f ten Fin f rierbarkeit der Ejakulate von zwei Bullen wurden Veränderungen der Spermatozoen im Bereich des Akrosoms festgestellt, die bereits beim Frischsamen, vermehrt aber nach dem Einfrieren nachzuweisen sind. Diese Veranderungen (Schwellungen, Ablösungen, Brüche) entsprechen denen, die im Frostsperma von Tierarten gefunden werden, deren Ejakulate generell schlecht einfrierbar sind (Eber, Schafbock, Chinchilla). Bei einem Bullen, dessen Ejakulate trotz guter Beweglichkeit nicht einfrierbar waren, konnten Hinweise für feinste Schädigungen des Membransystems als Ursache gefunden werden. Contents Spermatozoa from bulls whose ejaculates were well frozen, partly frozen and unfreezable were examined in the electron microscope before and after freezing. The ejaculates of two bulls which were imperfectly freezable had spermatozoa which showed consistent damage to the acrosome complex. The alterations (swelling, detachment, rupture) were seen in fresh semen and to a greater extent in semen frozen in liquid nitrogen and then thawed. They correspond to the changes found in the frozen semen of these species of animals, whose ejaculates are in general poorly freezable (rain, boar, chinchilla). In the ejaculate of one bull which was not freezable in spite of good motility the cause was found to be slight damage to the membranous system.     

SEMEN WASTAGE AT A BOVINE ARTIFICIAL BREEDING CENTRE

The consequences of the use of swirl assessment at a bovine artificial breeding centre were studied. Data relating to the amount of semen discarded, fertility of accepted semen, the relationship between discard rates and age of bull and ejaculate number have been presented. The overall differences in fertility between semen with swirl ratings, 5, 6 and 7 were very small. About 50% of all ejaculates collected in the period considered were discarded at the first inspection. The discard rate tended to be greater in young and in older bulls. The discard rate was significantly greater (P less than or greater to 0.001) for second ejaculates. It was considered that a proportion of rejected semen may have been of good fertility. Semen wastage after freezing was low.