Fertility of cryopreserved sperm in three bulls with different Robertsonian translocations

The fertility of three bulls carrying different Robertsonian translocations (rob(1;29), rob(14;17) and rob(26;29)) was evaluated. Oocytes-cumulus complexes obtained from slaughterhouse-derived ovaries were matured and then fertilised in vitro with frozen/thawed seminal material from the above mentioned subjects, and from control bulls with normal karyotype. An assessment was first made of the concentration, vitality and acrosome integrity of the seminal material to be sure that possible differences in the results of the in vitro fertilisation experiments were not due to seminal material quality. The results of the experiments, evaluated by the percentage of cleaved embryos and blastocysts per cleaved embryo, indicated that the three bulls carrying Robertsonian translocations had similar fertilising power and semen qualitative parameters to the controls. These data suggest that neither gametogenesys impairment nor decreased spermatozoa fertilising capacity is responsible for the reduced fertility in bulls with Robertsonian translocations. What the data do confirm is that the observed in vivo hypofertility for karyologically abnormal bulls is mainly due to early embryonic mortality.     

Herapin-binding proteins of canine seminal plasma

Heparin-binding proteins (HBP) from seminal plasma have been expected to participate in modulation of the acrosomal reaction, and have been correlated with fertility in some species. However, they have not been described in the dog. The aim of this study was to document the HBPs of canine seminal plasma. Six pooled samples of seminal plasma from three crossbred dogs were used. The HBPs were isolated by heparin affinity chromatography and the fractions recovered were pooled. One-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was carried out on 12 and 18% vertical minigels. The stained gels were scanned and the molecular weight (kDa) values for each band within a lane were calculated by image analysis software. The electrophoresis analysis of the pooled eluded fractions identified 19 bands, with molecular weights varying from 61.5 to 5.2 kDa. Previous studies, using one-dimensional SDS-PAGE, identified two bands (67 and 58.6 kDa), which were positively correlated with some semen parameters (sperm motility, sperm vigor, percentage of morphologically normal sperm and plasma membrane integrity). The 61.5 kDa band detected in the present study apparently corresponded to the 58.6 kDa band identified previously. Canine seminal plasma contained HBP; since HBP modulate the acrosome reaction in other species, they may have the same function in the dog. Further studies are necessary to better characterize this protein and determine if it is associated with fertility in the dog.     

Altitude, migration, and fertility in the Andes

In order to examine the relationship between hypoxia and reduced fertility of high Andean populations, a sample of 241 females living in the low-altitude Tambo Valley of Peru was studied. 63 of the subjects were born in the low-altitude valley, 121 were migrants from high altitudes, and 57 were migrants born in low altitudes. The rate of abortion was low among high-altitude subjects before they migrated, but became greater after migrating. It was found that the high-altitude populations had almost twice as long parity intervals than the low-altitude populations. Compared to migrants born at low altitudes, the high-altitude-born subjects who migrated to low altitudes had higher fertility rates. The results of the study are consistent with the hypothesis that high altitudes, through anoxia, have a lowering effect on fertility. Of the several possible explanations which might account for the increase in fertility of downward migrants on migration from high to low altitude (migration, socioeconomic factors, acculturation, seasonal male emigration from high altitude, and removal of hypoxia stress), altitude appears to be the most significant.     

Description and validation of a novel technique to study the bacterial flora of the teat duct of ewes

We describe a new technique that may be used for studying the bacterial flora of the teat duct in live ewes, and we present evidence regarding its accuracy, safety and repeatability. A sterile, plastic, 20 G catheter is used for sampling. The catheter stylet is taken out and the plastic catheter cut with a sterile blade to a length of 2 mm. In order to ensure accurate and consistent cutting of the catheter at the desired length, a sterilized ruler is placed beside the catheter. The whole procedure is carried out under aseptic conditions, on sterilized surgical screens. Before sampling, a thorough disinfection with povidone iodine scrub solution of the teat apex and the lower (1 cm) part of the teat skin is carried out. The catheter is held by the investigator from the cannula hub; then, it is inserted into the teat, rolled around the internal teat wall, in order to sample the mucosa and then withdrawn. Material collected on the tip of the catheter is plated onto bacteriological media. The sensitivity of the technique compared to a method that detects the bacterial flora of the teat after slaughter of the animals (scratching of the teat mucosa by means of a scalpel blade) was 99.1% for field samples and 100% for experimental samples; specificity of the technique was 90.7 and 100%, respectively, kappa coefficient between results of the two methods was 0.9045. The safety of the technique was assessed in an investigation, where ewes were repeatedly (eight times) sampled by means of this technique, throughout their lactation; in no case was clinical or subclinical mastitis recorded. Finally, the repeatability of the method was assessed in 37 paired samples, obtained 30–40 min apart; a 100% match was recorded between bacteriological results of the two samplings of each pair. We conclude that the method can be useful in research studies, in order to obtain information regarding bacterial ecology and dynamics inside the teat of ewes.     

Hemoglobin genotype, fertility, and the malaria hypothesis

Epidemiological and clinical studies (Fleming et al. 1985; Perrin et al. 1982) indicate that hemoglobin (Hb) AS individuals have a selective advantage in malarial environments. Thus the high frequency of Hb S in human populations has been attributed to the decreased malarial morbidity and mortality experienced by Hb AS heterozygotes. It has also been suggested that Hb AS women have a higher fertility than that of Hb AA women, thus contributing to the elevated frequency of Hb S in malarial environments (Livingstone 1957). Firschein (1961) demonstrated a significantly greater fertility among Hb AS females, whereas Custodio and Huntsman (1984) documented no fertility differential between Hb AS and Hb AA women. Here I examine the reproductive careers of Hb AA and Hb AS subjects 40 years of age and older from Limon, Costa Rica. The purpose is to determine whether normal homozygotes and heterozygotes have significantly different fertilities. The research shows that these groups do not have significantly different completed family sizes (t = 0.38, ns) or significantly different numbers of pregnancies (t = 0.34, ns), live births (t = 0.36, ns), or abortions (t = 0.20, ns). My results support previous suggestions that differential fertility does not contribute to the maintenance of the Hb S polymorphism.     

37种杜鹃花属植物在迁地保育下的自然授粉研究

该文对主要栽培在四川都江堰龙池基地(海拔1 700 m)及玉堂基地(海拔700 m)和峨眉山生物试验站(海拔805 m)的5亚属15亚组37种杜鹃花属植物的自然授粉开展了为期4 a的数据采集与研究。以绿苗率(Gs)为主要指标,绿苗系数(Gc)、坐果率(St)和单位可育种子数(Sf)为辅助指标,初步揭示了迁地保育条件下杜鹃花属植物自然授粉的育性适应及其特征。结果表明:(1)除黄花杜鹃(Rhododendron lutescens)未形成种子外,受试的其他36个杜鹃花种类均能在其保育地点上不同程度地完成从种子(幼苗)到种子的生命循环,其中高可育型24种、可育型11种、弱育型1种。(2)上述4项指标,尤其是保育条件与原生地条件下同种的绿苗率、绿苗系数和单位能育种子数比较,均能不同程度地反映育性适合度的差异情况。(3)在上述具有不同程度可育性的36种杜鹃花中,有24种有不同程度的败育现象,其成因可能为不同程度的自交和花期重叠的同亚组到不同亚组异种间自然交配所引起的遗传不适,由于遗传选择的限制或胁迫,这种现象在一些开花个体有限的种类中尤为突出,因此保证最小存活种群(the minimum viable population,MVP)对于该属植物迁地保育至关重要。(4)杜鹃花属的特定种类可能存在一个受种性制约的单位种子数量幅度和上限,即不同的类群与种类的单位种子数量存在差异。(5)为探索自交和种间杂交提供了认识起点和参照,并提出了有关杜鹃花属植物可育性综合评价的指标体系和方法。     

The removal of the seminal vesicles from the boar and the effects on the semen characteristics.

A technique is described for the removal of the seminal vesicles from the boar. The operation was carried out on twelve animals and six of the animals were subsequently trained for semen collection. The seminal plasma from the boars after surgery compared with normal litter mates had a more watery consistency and did not form the characteristic gelduring ejaculation. The sperm concentration was 49% lower while the total reduction of sperm number ejaculate was 78% in the experimental animals, but the ratio of living to dead spermatozoa remained unchanged. The concentrations of citrate and protein were significantly depressed in the seminal plasma of the animals after surgery and the pH increased; the osmolarity remained unchanged. In semination of gilts with the semen from experimental boars revealed no significant loss of fertility compared with the normal controls. Animals slaughtered up to 17 months after surgery showed no regeneration of the seminal vesicles.     

Seminal plasma adipokines: involvement in human reproductive functions

Infertility, which increased worldwide over the past few decades, has recently been linked to obesity prevalence. Adipokines, produced by adipose tissue, could be the link between obesity and infertility. The association between circulating adipokines and female infertility has been extensively studied in the last ten years. However, the male aspect has been less investigated, although some adipokines are present in seminal plasma. We have attempted to analyze published studies that measured seminal plasma adipokines and their relationships with semen parameters. Apart from leptin, other seminal adipokines have rarely been studied. Indeed, leptin seems to have a differential role depending on its concentration in the seminal plasma. Thus, it could have a beneficial effect at lower concentrations but a deleterious effect at higher seminal levels. Although some studies are currently available, the roles of leptin and other adipokines in seminal plasma on sperm parameters and their consequences on male fertility remain to be clarified.     

Heparin-binding proteins of human seminal plasma: purification and characterization

Human seminal plasma (HuSP) contains several proteins that bind heparin and related glycosaminoglycans. Heparin binding proteins (HBPs) from seminal plasma have been shown to participate in modulation of capacitation or acrosome reaction and thus have been correlated with fertility in some species. However, these have not been studied in detail in human. The objective of this study was to purify major HBPs from HuSP in order to characterize these proteins. HBPs were isolated by affinity-chromatography on Heparin-Sepharose column, purified by reverse-phase high-performance liquid chromatography (RP-HPLC) and Size-exclusion chromatography and checked for purity on sodium-dodecyl PAGE (SDS-PAGE). Identification of HBPs was done by matrix-assisted laser desorption-ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS). Here we report the purification and identification of seven HBPs in seminal fluid. The major HBPs are lactoferrin and its fragments, semenogelin I fragments, semenogelin II, prostate specific antigen, homolog of bovine seminal plasma-proteins (BSP), zinc finger protein (Znf 169) and fibronectin fragments. In this study we are reporting for the first time the purification and identification of BSP-homolog and Znf 169 from HuSP and classified them as HBPs. Here we report the purification of seven clinically important proteins from human seminal fluid through heparin affinity chromatography and RP-HPLC, in limited steps with higher yield.     

Selenium status of idiopathic infertile Nigerian males

Selenium concentration in the sera and seminal plasma of 60 infertile males (40 oligospermia and 20 azoospermia) and 40 males with proven evidence of fertility (normospermia; control group) were estimated using atomic absorption spectrophotometry. Results were correlated with spermatogram and hormonal levels in order to determine their relationship and significance in male infertility. The mean serum concentrations of selenium was found to be significantly increased in oligospermic compared to azoospermic subjects and controls (p<0.01), whereas the seminal plasma level was significantly higher in azoospermic compared to oligospermic subjects and controls (p<0.001). Thus, the ratio of serum selenium to seminal plasma selenium was 1∶1 in controls, 4∶1 in oligospermia, and 1∶2 in azoospermic subject. A significant inverse correlation was observed between serum selenium level and sperm count (p<0.01). Similarly, seminal plasma selenium correlated with spermatozoa motility, viability, and morphology. Serum selenium level shows positive correlation with the serum testosterone level (p<0.01). In conclusion, there appears to be a physiological balance in the distribution of selenium in serum and seminal plasma compartment of control males. A disturbance in this balance has a significant influence on spermatogenesis. Selenium appears to have a positive influence on Leydig cells, thus influencing the secretion of testosterone.     

Influence of seminal plasma on the fecundity of chicken spermatozoa

This study was undertaken to investigate the influence of seminal plasma on the fecundity of chicken sperm. Sperm diluted with either incubated seminal plasma (5 or 37 degrees C for 24 h) or seminal plasma from incubated whole semen (5 or 37 degrees C for 24 h) had lower fertility levels and motility scores than sperm diluted in either fresh seminal plasma or a synthetic diluent. The number of sperm with damaged membranes increased with seminal plasma derived from 37 degrees C incubation. The depressive effect of incubated seminal plasma on semen fertility was eliminated by microfiltering .(0.22 mum) the seminal plasma either before or after incubation. Filtration of seminal plasma was only effective in eliminating the depressive effect on sperm motility when filtering was done after incubation. Filtration of seminal plasma reduced the percentage of damaged sperm in all treatments. It can be concluded that there are factors in seminal plasma that are deleterious to the fecundity of chicken spermatozoa and they may be derived from degenerating sperm and/or various fluids, cells and debris collected with the semen during manual semen collection.     

The impact of blood and seminal plasma zinc and copper concentrations on spermogram and hormonal changes in infertile Nigerian men

Zinc (Zn) and copper (Cu) concentrations in sera and seminal plasma of 60 infertile males (40 oligozoospermic and 20 azoospermic) and 40 males with evidence of fertility (normozoospermic; controls) were estimated using atomic absorption spectrophotometry. The results were correlated with the subject's spermogram and hormonal levels in order to determine their relationship and significance in male infertility. The mean serum concentration of zinc was significantly (p<0.01) higher in oligozoospermic males when compared to azoospermic subjects and controls. The ratios of serum Zn to seminal plasma Zn were 1:1, 1:3 or 1:4 in oligozoospermic, normozoospermic or azoospermic subjects, respectively. While the mean Cu concentration was significantly higher in serum than seminal plasma in all groups, the Zn concentration was significantly (p<0.05) higher in seminal plasma than serum. The Cu/Zn ratio in seminal plasma was significantly (p<0.01) higher in controls compared with other groups. A significant (p<0.01) inverse correlation was observed between serum Zn and sperm counts. Similarly, seminal plasma Zn negatively correlated with spermatozoa viability. In conclusion, the measurement of serum Zn level, apart from being a good index of the assessment of prostatic secretion and function, may be considered a useful tool in addition to other parameters in assessing male infertility. Also, a lower Cu/Zn ratio in seminal plasma may serve as a supportive tools in assessing male infertility.     

Comparison of two skin antiseptics in blood transfusion. Comparison with the results of the bacteriologic control of labile blood products

The comparison of two skin antiseptics (70 degrees alcohol and 0,5% alcoholic chlorhexidine solution) was carried out by the method of in vivo impressions. The study used 45 healthy volonteers from whom a bacteriological sample was taken from both bends of the elbow, without use of an antiseptic, and after the application of one according to usual sample taking methods. The subjects were divided into two sets according to the antiseptic being tested. The results of the cultures after 24 and 48 hours are expressed in the number of germs per cm2, which thus permits us to calculate a reduction in percentage or log 10 form, and to appreciate the efficiency of the antiseptic studied. Routine bacteriological inspection of labile blood products was carried out on products picked at random every week during 18 months of exclusive use of 70 degrees alcohol by the sample taking services. The results show a comparable in vivo effectiveness of the two antiseptics on the superficial aerobic flora of the bend of the elbow, and the ease of use of 70 degrees alcohol has led to its choice. This choice is confronted with the results of the inspection of blood products: out of 1 293 inspections, only one slight contamination (cocci gram+) was found on a unit of whole blood. These data as a whole can be compared with the work of various authors.     

Assessing the quality of raw semen: a review

An analysis of semen characteristics can provide a reasonable basis upon which to develop a strategy for maximizing the fertility of a stallion. However, the repeatability of semen characteristics between ejaculates within stallions is low to moderate. Factors such as season, collection technique, frequency of collection and disturbances in spermatogenesis contribute to this variation. Fertility can, however, be influenced by a host of other factors besides semen characteristics, including management, semen handling procedures and the number of mares being bred. Parameters usually included in a conventional evaluation of raw semen quality are volume, sperm concentration, total number of spermatozoa in the ejaculate, percentages of motile spermatozoa, sperm morphology, seminal pH, longevity of sperm motility and bacteriological status. Although, these evaluations provide a lot of information, their correlations with fertility are somewhat conflicting. However, it seems likely that the prediction of male fertility could be improved if additional parameters based on the functional characteristics of spermatozoa were to be used. Several functional tests have been investigated, such as the use of fluorescent stains as a marker for cell membrane integrity, sperm-oocyte binding tests and the hypoosmotic swelling test. In this study, emphasis is placed on sperm motility and sperm morphology, but in addition, some functional tests are also discussed.     

Sterile microenvironment in prevention of wound infection.

A prospective controlled trial was carried out to assess the effect of using a wound isolator on reducing postoperative infection. A total of 291 patients undergoing hip pinning for fractures of the neck of femur entered the trial and were allocated at random to have their wound contained in a wound isolator (study group) or dressed with a standard gamma-irradiated adhesive dressing (control group). The bacteriological flora of the patient was monitored before, during, and after operation and that of the ward before and after. No significant difference was found in the flora of the wards in which the patients were nursed. On several occasions the source of the infective organism was traced to the ward but never to the theatre. The isolator prevented direct contamination and airborne cross-infection of the wound and appreciably reduced the rate of infection.     

Effect of trace elements on the seminal oxidative status and correlation to sperm motility in infertile Saudi males

The impact of trace elements, especially zinc, selenium, copper, and magnesium, on male fertility has gained great interest and significance. Increased oxidative stress and altered trace element levels are probable etiological factors underlying male reproductive dysfunction and infertility. The present study focused on the evaluation of seminal oxidative markers, such as reactive oxygen species (ROS), malondialdehyde (MDA), and total antioxidant capacity (TAC), and trace element levels in the normozoospermic fertile control group (n = 40) and asthenozoospermic infertile group (n = 30). Semen from infertile men exhibited significantly higher ROS and MDA levels accompanied with significant decline in TAC and trace element (zinc and magnesium) levels. Furthermore, a significant correlation was observed between trace elements and oxidative markers with sperm motility. The current study revealed increased lipid peroxidation and oxidant-reductant imbalance that leads to deterioration of semen quality and male infertility. Thus, oxidative stress and trace elements can be considered important biomarkers of male infertility. Measurement of seminal oxidative stress with conventional seminological parameters must be integrated in fertility assessment from early stages to ensure healthy semen characteristics and fertility in men.     

The Atlantic elements in the Swiss flora: distribution, diversity, and conservation status

The purpose of this study was to examine the composition, distribution, ecology, and conservation status of the Atlantic elements of the Swiss flora. About 195 Atlantic and 80 Mediterranean–Atlantic vascular plant species of the European flora have been used as the basis for our analysis. The complete list of 3,143 taxa has been used as the reference for the Swiss flora. The distributions of the species are illustrated in coincidence maps based on the computer database of the Data Centre of the Swiss Flora in Geneva, Switzerland. Our study demonstrates clearly that the Atlantic flora of Europe requires a new biogeographical appraisal. The Swiss flora comprises 66 Atlantic and Mediterranean–Atlantic taxa, which are taxonomically and ecologically highly diverse. Switzerland contains 44% of all European Sub-Atlantic plants. This confirms the Sub-Atlantic geographical position of Switzerland. Only one Eu-Atlantic species growing in Switzerland, Vicia orobus, can be classified as native with certainty. This species is critically endangered and merits the highest conservation priority. Although a very alpine country, Switzerland has a relatively large number of Mediterranean–Atlantic species. The Atlantic and Mediterranean–Atlantic plants are a very threatened group in Switzerland, with wetland plants the most imperilled ecological group. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Role of fluid from seminal vesicles and coagulating glands in sperm transport into the uterus and fertility in rats.

The relationship between the quantity of seminal vesicle secretion in the ejaculate, the percentage of spermatozoa reaching the uterus and fertility was studied in rats. Different portions of seminal vesicles were removed from male rats; 15 min after coitus (day 0), the numbers of spermatozoa in the uterus and vagina were counted and the vaginal plug characteristics were noted. Fertility was evaluated by the number of fetuses on day 14. A gradual decrease in the percentage of spermatozoa in the uterus was positively related to the reduction in seminal vesicle secretion, estimated by plug weight. This decline was not caused by a delay in sperm transport to the uterine lumen and the results suggested that the spermatozoa that fail to enter the uterus in the first minutes after coitus never enter. The vaginal plug weight, which is related to the seminal vesicle weight, and the position of the plug, which must be firmly lodged into the cervical opening, seem to be the most important conditions for promoting the rapid passage of spermatozoa into the uterus. When the seminal vesicles were partially removed, the plug was not tightly lodged and formed a 'cup' filled with spermatozoa. The number of fetuses did not show a close correlation with the quantity of seminal vesicle secretion. Studies of males in which the seminal vesicles had been removed indicated that a normal number of fetuses can be obtained despite low numbers of spermatozoa reaching the uterus. Ablation of the coagulating glands showed that, when there is no vaginal plug, no spermatozoa reach the uterus and fertility is suppressed. Nevertheless, the complete removal of coagulating glands is difficult; when small portions of these glands remain, the vaginal plug is formed and then fertility is achieved.     

Effect of season on bovine seminal plasma proteins in Thailand

Although season has been shown to affect bull sperm quality and fertility in some studies, the effect of season on seminal plasma proteins has not been examined. In the present study, seminal plasma proteins were analysed by Fast Protein Liquid Chromatography (FPLC), to separate the phosphorylcholine-binding proteins and heparin-binding proteins from the other proteins. Semen samples were collected from bulls in three seasons: winter, summer and the rainy season. Sperm quality was analysed by flow cytometry and computer assisted sperm analysis, and further aliquots of semen were used to prepare the seminal plasma for FPLC. Meteorological data were available from a location close to the bull station. There were slight differences in sperm kinematics between seasons, but other parameters of sperm quality were not different. Minor differences in the phosphorylcholine-binding proteins were detected according to season, being lower in summer than in winter or in the rainy season, although there were no changes in the heparin-binding proteins. Temperature, humidity and rainfall differed between winter and the rainy season, but no differences were observed between summer and the rainy season except in the temperature humidity index (THI). However, the THI was above the threshold indicative of heat stress in all seasons, which could explain why few seasonal differences in protein composition were detected in this study. Alternatively, the bulls could have been well-adapted to heat stress. In conclusion, there were only slight differences in bull sperm quality and seminal plasma proteins between seasons during this study.