Effect of semen collection method (artificial vagina vs. electroejaculation), extender and centrifugation on post-thaw sperm quality of Blanca-Celtibérica buck ejaculates

The aim of this study was to evaluate the effect of semen collection method (artificial vagina compared to electroejaculation), season in which the semen was collected (breeding season compared to non-breeding season), freezing extender (Biladyl(?), Andromed(?) and skim milk based extender) and pre-treatment procedure (washing compared to non-washing) on post-thaw semen quality in buck. Ejaculates from seven bucks of the Blanca-Celtibérica breed were collected by artificial vagina and electroejaculation during the breeding (July to December) and non-breeding season (January to June). Samples were split in two aliquots and one of them was washed. Three freezing extenders were evaluated on washing and non-washing sperm samples. Ejaculates collected by artificial vagina had a greater sperm quality after thawing, with greater values (P≤0.05) for SM (sperm motility), NAR (acrosome intact), YO-PRO-1-/PI- (intact spermatozoa), and Mitotracker+/YO-PRO-1- (spermatozoa with active mitochondria) and lower % DFI (DNA fragmentation index). Thawed sperm samples which were collected during the breeding season had greater values (P≤0.05) for NAR, intact spermatozoa and spermatozoa with active mitochondria, than those semen samples obtained during the non-breeding season. Semen freezing with Biladyl(?) and Andromed(?) resulted in a greater sperm quality (P≤0.05) after thawing in relation to milk-based extender. Washing procedure had no effect on sperm parameters assessed at thawing. Results from the present study suggest that the success of semen cryopreservation in Blanca-Celtibérica goat depends on semen collection method and season, as well as on the extender used. Thus, the post-thaw sperm quality will be greater (P≤0.05) when samples are collected by artificial vagina during the breeding season and when Biladyl(?) or Andromed(?) are used as freezing extenders.     

Quality of stallion semen obtained by a new semen collection phantom (Equidame) versus a Missouri artificial vagina

A study was performed to test a new semen collection device (Equidame phantom) that fractionates the ejaculate by comparing the quality of semen obtained by the Equidame phantom with that obtained by a Missouri artificial vagina. Semen from 4 Finnhorse stallions was collected 4 times per stallion by both methods. Half of the ejaculate was frozen and the other half extended and loaded into 2 Equitainer transport containers (24- and 48-h samples). Motility parameters were determined by a Hamilton-Thorn motility analyzer after cooled storage for 24 and 48 h and again after freezing/thawing. Raw and chilled semen samples were cultured and the number of bacterial colonies counted after incubations of 24 and 48 h. After a 24-h incubation the number of colony-forming units (CFU) in raw semen was significantly higher (P<0.01) when collected by the Missouri artificial vagina than by the Equidame phantom. After cooled storage, 75% of the semen samples contained no bacteria after an incubation of 24 h, and 69% yielded no growth after 48 h. The sperm-rich fractions (Cup 2) collected by the Equidame phantom had lower mean volumes (22.1 +/- 2.3 mL [+/- SEM] versus 101.6 +/- 9.3 mL) and significantly higher mean sperm concentrations (218.0 +/- 25.8 x 10(6) vs 86.2 +/- 8.1 x 10(6) cells/mL; P<0.05) than the total ejaculates collected by the Missouri device. The total and progressive motility of chilled and frozen-thawed semen did not differ significantly between collection methods. The Equidame phantom yielded semen that was of a lower bacteriological colony counts, but had sperm motility similar to that of semen collected with the traditional method by the Missouri artificial vagina.     

青岛崂山青山湾人工鱼礁区及附近海域鱼卵仔稚鱼种类组成与数量分布

为研究人工鱼礁对产卵鱼类的诱集效果和庇护效应,2014—2015年对青岛崂山青山湾海域人工鱼礁区及附近海域鱼卵和仔稚鱼的种类组成和数量分布进行了水平和垂直拖网调查. 2014年春、夏、秋3个季节的7个航次调查中,共采集鱼卵7306粒、仔稚鱼52尾,隶属于4目9科11属12种;2015年同期进行的13个航次调查中,共采集鱼卵10373粒、仔稚鱼159尾,隶属于6目11科14属15种.2年间鱼卵和仔稚鱼样品均以鲈形目数量最多,鲽形目次之,其中鱼卵样品中仅有少鳞鱚和短吻红舌鳎构成优势种;仔稚鱼样品以少鳞鱚优势度最高,但不存在仔稚鱼优势种.鱼卵和稚鱼的高优势度物种的季节间演替明显.春、夏、秋季鱼类浮游生物群落Margalef丰富度指数、Shannon多样性指数和Pielou均匀度指数平均值均较低,说明该海域鱼类浮游生物群落结构稳定性较低.2年间所采集物种对应的成鱼平均营养级分别为3.71和3.78,均属第三营养级中的低级肉食性鱼类,绝大多数为暖温种,无冷温性种类,表现出明显的暖温带特点.综合分析认为,人工鱼礁区鱼类浮游生物群落的多样性相对较高,这与鱼礁区流速和流场特征及复杂的空间异质性有关.     

Effectiveness of an artificial substrate for sampling macroinvertebrates in small streams

SUMMARY. Factors affecting invertebrate colonization of samplers consisting of twenty-five 5-cm diameter porcelain balls were studied in small northern California streams. Peaks in diversity, in number of individuals and in taxa, occurred after 2—4 weeks in each of four 10-week colonization runs. Most variation in number of organisms was correlated with increases in organic detritus. Time was the second most important variable.
The average coefficient of variation in total numbers collected from the artificial sampler was 0.40, slightly less than the average 0.56 for the Surber sampler. Published coefficients of variation for artificial substrates range from 0.109 to 0.849, which are similar to values reported from work with the Surber sampler. We had several practical difficulties with the artificial substrates. Samplers were lost, became clogged or buried, supported vertebrate predators and were costly. The samplers also collected some organisms in greater proportion than their occurrences in the natural substrate. In sampling small streams these disadvantages (compared to a Surber sampler) outweigh the slight reduction in sampling variability.     

Studies on camel semen. I. Electroejaculation and some aspects of semen characteristics

A method for collection of camel semen by electroejaculation has been described for the first time. The volume of the ejaculates obtained was slightly lower than that obtained using an artificial vagina. The sperm count was considerably higher, but other parameters including pH, percentage of live sperm and sperm dimensions were comparable to those of ejaculates collected by the artificial vagina. The camel spermatozoon is generally smaller than those of other animals. The head is somewhat elliptical in shape and slightly tapering at the base. Mean sperm dimensions for the head, middle piece and tail in μm were 5.5, 6.9 and 35.6 respectively; total sperm length was 48.0 μm.     

Effect of the collection method on semen characteristics of Zebu and European type cattle in the tropics

The effect of the collection method on the characteristics of fresh semen and the recovery of spermatozoa after thawing was studied in 30 Zebu bulls (Bos indicus ) and in 30 Brown Swiss (Bos taurus ) bulls. Semen was collected by using an artificial vagina and by electroejaculation; the ejaculates were individually evaluated. Semen was diluted for freezing in skimmed milk and stored in 0.5-ml French straws, at a concentration of 30 x 10(6) spermatozoa. Data were evaluated by analysis of variance using a factorial model which included collection method, breed effect, and interaction between method and breed, with each bull as a block. Higher volume and pH of the semen was obtained following electroejaculation. Conversely, higher concentration prior to freezing and better progressive motility after thawing was observed in semen collected with an artificial vagina. No differences in motility were obtained in fresh semen between methods. Better post-thaw recovery occurred when the semen was collected by an artificial vagina, independently of the breed type.     

Characterization of semen collected from beagles and captive Japanese black bears (Ursus thibetanus japonicus)

This study characterized semen collected from the Japanese black bear, Ursus thibetanus aponicus, to provide information on semen cryopreservation for artificial breeding. Preliminary studies using a beagle dog as the model species showed that sperm concentration and total sperm count were lower in semen collected by electroejaculation than in semen collected by digital manipulation, but that sperm motility, viability and morphology were similar. Characterization of semen obtained from Japanese black bears by electroejaculation under general anesthesia revealed that semen volume and total number of spermatozoa collected were lower; but that sperm concentration, motility, viability and morphology were equivalent to those reported in other ursids. When semen was collected via a catheter inserted into the urethra during the stimulation for ejaculation, the sperm concentration, total sperm count and motility were relatively higher than when semen was collected directly in a test tube. Specific normal semen characteristics (mean +/- SEM) were pH, 7.6 +/- 0.0; volume, 0.212 +/- 0.038 mL; sperm concentration, 361 +/- 100 x 10(6)/mL; total sperm count, 84.0 +/- 32.2 x 106; +++ motility, 30 +/- 5%; motility, 77 +/- 3%; viability 77 +/- 2%; and abnormal morphology, 11+/- 2%. These results suggest that semen can be collected from Japanese black bears by electroejaculation.     

Seed germination for an annual form of Zostera marina from the sea of Cortez,Mexico

Seed of Zostera marina L. collected at Punta Chueca, Sonora, that was germinated in artificial seawater under growth chamber conditions was less affected by salinity than by temperature. Mean germination was higher for seed collected on reproductive shoots in situ (43%) than for seed collected in fresh beach debris (17%), but no germination was recorded for seed collected in dried beach debris. Mean germination for seed kept at either 15 or 35‰ salinity was approximately equal. Earliest germination was recorded in late April 1980, two weeks after collection, and germination continued through March 1981 at 18–20°C, conditions which are comparable to winter water temperatures in Canal del Infiernillo. Germination was inhibited at 28–32°C, temperatures that are near summer water conditions. The germination responses reflect the adaptive strategies of an annual population to habitat conditions near the southern limit of the species.     

Effects of liner composition, cooling profile and time of glycerolization on survival of bovine spermatozoa

Bovine spermatozoa from five bulls (158 ejaculates) were collected into one-piece combination liner-collection cones in an artificial vagina. Spermatozoal motility and differential staining were evaluated at each sequential step of semen processing. Linear 30-min and nonlinear 150-min cooling profiles resulted in equivalent postthaw spermatozoan survival rates. Preglycerolation holding times of 60 and 150 min, after 30-min linear cooling, were associated with increased, but nonsignificant, postthaw survival rates for bovine spermatozoa collected in polyethylene liner-collection cones. Highly significant differences (P<0.0001) were found to be associated with the composition of the liner-collection cones used. Motility estimates made at collection and after thawing from liquid nitrogen storage indicated that at least 50% of motile spermatozoa collected in polyethylene liner-collection cones survived; whereas, not more than 30% of the cells survived when they had been collected in rubber liner-collection cones.     

Functional parameters of chimpanzee (Pan troglodytes) sperm from ejaculates collected by rectal probe electrostimulation and by artificial vagina

This study compares functional parameters of sperm from ejaculates collected from 15 adult male chimpanzees using rectal probe electrostimulation (RPE) and from 10 adult male chimpanzees trained to use an artificial vagina (AV). Computer assisted motion analysis (CAMA) showed no significant differences in mean values for straight line velocity (VSL), linearity (LIN), curvilinear velocity (VCL), and lateral head movement (ALH) of sperm from ejaculates collected by RPE and by AV. There was, however, a significant difference (P < 0.01) in the population distribution for VSL and LIN, which indicates that sperm swim in a more convoluted manner in ejaculates collected by RPE than in ejaculates collected by AV. In the hamster zona-free ovum penetration assay (SPA), there were no significant differences in the percentages of hamster oocytes penetrated by sperm or in the number of sperm which penetrated each oocyte after 4 or 24 h incubation using sperm from ejaculates collected by RPE and by AV. Therefore, the lack of success using sperm from ejaculates collected by RPE to initiate pregnancy in the chimpanzee does not appear to result from abnormalities in sperm fertilizing capacity as measured in SPA. © 1996 Wiley-Liss, Inc.     

In vivo release of endogenous dopamine, 5-hydroxytryptamine and some of their metabolites from rat caudate nucleus by phenylethylamine

The in vivo release of endogenous 3,4-dihydroxyphenylethylamine (DA) and its metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 3-methoxytyramine (3-MT), and of 5-hydroxytryptamine (5-HT) and its metabolite, 5-hydroxyindoleacetic acid (5-HIAA), has been measured in the caudate nucleus of the anesthetized rat. A push-pull cannula was implanted into the brain, and the tissue perfused with artificial CSF or artificial CSF containing 5×10^–4 M phenylethylamine. The perfusate was collected and analyzed for DA, 5-HT and their metabolites by high performance liquid chromatography with electrochemical detection (HPLC-ECD). DA was released by phenylethylamine at rates significantly greater than its basal rate. 3-MT and 5-HT were undetectable in perfusates collected under basal conditions, but could be detected readlly during phenylethylamine stimulation. DOPAC, HVA and 5-HIAA concentrations were not significantly affected by phenylethylamine. The results suggest (1) that phenylethylamine may exert its behavioural effects through increased release of both DA and 5-HT, and (2) that in vivo measurements of the acid metabolites alone may not be indicative of the release of the amines.Special Issue Dedicated to Dr. Abel Lajtha.     

不同三尖瓣成形术治疗风湿性二尖瓣病变合并三尖瓣关闭不全临床研究

目的:比较行不同成形术治疗风湿性二尖瓣病变合并功能性三尖瓣关闭不全的外科疗效。方法:选取风湿性二尖瓣病变合并功能性三尖瓣关闭不全患者119例,按照治疗方法将患者分为对照组、三尖瓣人工环植入成形术组(成形环组)以及三尖瓣缝线成形术(缝线组),分别统计患者年龄、性别、手术方式、术前及术后心功能分级等指标,采用t检验对患者术前、术后2周以及术后6个月心脏各腔内径进行统计学分析。结果:患者行三尖瓣人工环植入成形术以及三尖瓣缝线成形术治疗后,心脏各腔内径均明显缩小,成形环组患者术后心脏内径缩小最显著,行三尖瓣缝线成形术患者次之。术前成形环组左心房、右心房以及右心室内径较对照组扩大明显(P0.05);术前缝线组左心房、右心房以及右心室内径较对照组扩大明显(P0.05);术前成形环组与缝线组右心房、右心室内径组间无明显差异;术后2周以及术后6个月三组间左心房内径无明显差异(P0.05)。术后2周成形环组以及缝线组右心房以及右心室内径仍大于对照组(P0.05),术前成形环组与缝线组组间无显著差异。术后6个月成形环组右心房以及右心室内径较缝线组显著缩小(P0.05),成形环组和对照组间无明显差异。结论:治疗风湿性二尖瓣病变合并功能性三尖瓣关闭不全的方法中,三尖瓣人工环植入成形术效果优于三尖瓣缝线环缩术。     

Estimation of 3D rotation of femur in 2D hip radiographs

Femoral radiographs are affected by the degree of rotation of the femur with respect to the plane of projection. We aimed to determine the 3D rotation of the proximal femur in 2D radiographs. A 3D Statistical Appearance Model (SAM), which was built from CT images of cadaver proximal femurs (n=33) was randomly sampled to form a training set of 500 bones. Nineteen clinical CT images were collected for testing. All CT images were rotated to ±20° in 2° division around the shaft axis, ±10° around medial-lateral axis, and by simultaneous rotation of both axes (±16° and ±8° around shaft and medial-lateral axes). In each orientation, a 2D projection was recorded for generating a 2D SAM. The outcome parameters of the 2D SAM were used as input for a linear regression model and an artificial neural network to predict the rotation. The artificial neural network estimated the rotation more accurately than the linear regression. For artificial neural networks the mean errors were 4.0° and 2.0° around the shaft and medial-lateral axes, respectively. For an individual radiograph, the confidence interval of estimation was still relatively large. However, this method has high potential to differentiate the amount of rotations in two image sets.     

Non-invasive assessment of endocrine differences in day-old chicks (Gallus domesticus) by analysis of the immunoreactive oestrogen excreted in the egg

Oestrone, oestradiol and an uncharacterized polar fraction of oestrogenic substances were monitored by radioimmunoassay in Day-0 domesticated chickens (Gallus domesticus) after routine 21.5-day artificial incubation. Paired adrenals and gonads, blood serum and in-ovo excrement (allantoic urates plus cloacal expressions) were collected from 9 female and 10 male chicks. All samples were homogenized, hydrolysed and extracted with ether. Component oestrogens were isolated by celite:ethylene glycol column co-chromatography before assay. All egg excretory oestrogen measurements were higher (P less than 0.001) in females than in males. Oestradiol was the major excretory oestrogen in both sexes; oestradiol:oestrone ratios (mean +/- s.e.m.) in females and males were 18.45 +/- 2.42 and 3.03 +/- 0.37, respectively. Females had higher (P less than 0.01) total oestrogens for gonadal and adrenal homogenates and for blood serum samples than did the males; oestradiol was generally the major component.     

An extraordinary degree of structural specificity is required in neural phospholipids for optimal brain function: n-6 docosapentaenoic acid substitution for docosahexaenoic acid leads to a loss in spatial task performance

This study was conducted to determine whether provision of preformed dietary docosapentaenoic acid (DPAn-6) can replace docosahexaenoic acid (DHA) for brain function as assessed by spatial task performance. A newly modified artificial rearing method was employed to generate n-3 fatty acid-deficient rats. Newborn pups were separated from their mothers at 2 days of age and given artificial rat milk containing linoleic acid (LA), or LA supplemented with 1% DHA (DHA), 1% DPAn-6 (DPA) or 1% DHA plus 0.4% DPAn-6 (DHA/DPA). The animals were then weaned onto similar pelleted diets. At adulthood, behavioural tasks were administered and then the brains were collected for fatty acid analysis. The LA and DPA groups showed a lower (63-65%) brain DHA than the dam-reared, DHA and DHA/DPA groups and this loss was largely compensated for by an increase in brain DPAn-6. The brain fatty acid composition in the DPA group was the same as that in the LA group at adulthood. In the Morris water maze, the LA and DPA groups exhibited a longer escape latency than the dam-reared and DHA groups and had a defect in spatial retention. In conclusion, DPAn-6 could not replace DHA for brain function, indicating a highly specific structural requirement for DHA.     

Cortisol response in heifers to artificial insemination, natural mating, and no mating at estrus

Eight mixed-breed dairy heifers were used in a random block design to study the stress-producing effect of various types of mating on heifers. Treatments consisted of artificial insemination, natural mating, or not mating at estrus. Stress was evaluated by measuring blood plasma cortisol. Blood samples were collected by indwelling jugular catheters at 60 and 30 min before treatment; immediately before treatment (0 min); and 5, 15, 30, 60, 120 and 180 min after treatment. Means for cortisol were not significant for treatment or treatment across time. Mean cortisol response to the artificial insemination, natural mating and no mating treatments were 5.36, 8.05 and 5.56 ng/ml, respectively. These results suggest that the use of artificial insemination does not impose and added stress at estrus.     

西双版纳地区六种林型地表蜘蛛多样性比较研究

为探索地表蜘蛛多样性及其变化与森林类型和管理方式的关系, 在西双版纳勐仑自然保护区选择热带季节雨林、石灰山季节雨林和山地常绿阔叶林, 在自然保护区附近选择人工纯林、胶茶群落和橡胶林, 共6种林型, 每种林型选择3块样带, 共设置研究样地18块, 分别于2006年12月上旬（雾凉季）、2007年3月下旬（干热季）和2007年7月上旬（雨季）, 以单位地表面积法收集地表蜘蛛的物种组成和数量数据, 并以蜘蛛种类和数量分布为属性进行典范对应分析（CCA）, 探讨不同类型植被与地表蜘蛛多样性的关系。共采集蜘蛛标本9 849头, 用于统计分析的成熟蜘蛛3 119头, 归属于30科, 其中幽灵蛛科、皿蛛科、球蛛科和小密蛛科是地表蜘蛛的优势类群。各林型科的数量为: 热带季节雨林24科, 石灰山季节雨林22科, 山地常绿阔叶林22科, 人工纯林20科, 胶茶群落21科, 橡胶林19科; 各林型特有科数量: 热带季节雨林2科, 山地常绿阔叶林2科, 橡胶林1科; 而仅在雨林中分布的科4个（占全部30个科的13.3%）, 仅在自然林中分布的科6个（20.0%）, 仅在人工林中分布的科1个（3.3%）。从蜘蛛的数量分布看, 个体密度在热带季节雨林显著高于其他5种林型; 橡胶林多样性指数和丰富度指数显著低于3种自然林, 而均匀度指数的最低值也同样在橡胶林出现。CCA分析和聚类分析的结果表明, 6种林型趋于分成2组, 即: 自然林和人工林; 在自然林中两种次生林的相似程度更高; 人工林中人工纯林（非橡胶林）与胶茶群落的相似程度更高。以上结果表明: （1）森林砍伐后种植人工林措施改变了该地区地表蜘蛛群落的物种分布格局; （2）蜘蛛多样性随着人为干扰程度增加有减少的趋势; （3）减少人为干扰和增加植被群落多样性（橡胶林中种植茶树）对保护和恢复物种多样性有重要意义。