低氧分压通过低氧诱导因子(HIF)影响卵母细胞的体外成熟和早期胚胎的发育

卵母细胞和早期胚胎的体外培养(IVC)是哺乳动物胚胎工程的一项关键技术,是生殖生物学和转基因与克隆技术等生物技术研究的基础.影响卵母细胞和早期胚胎体外培养的因素包括培养液组成成分、离子浓度和渗透压,培养环境气相组成和温度等.最近研究表明,培养系统中的氧分压可以显著影响胚胎的发育,而这种影响主要由低氧诱导因子(hypoxia-inducible factor,HIF)进行调控.作者综述了氧分压及HIF对卵母细胞和早期胚胎体外发育的影响,并对HIF的结构及调控机制进行讨论,为研究低氧培养技术在卵母细胞和早期胚胎体外培养中的应用提供依据.     

Effects of Low Oxygen Tension for in vitro Oocyte Maturation and Early Embryo Development through Hypoxia-inducible Factor (HIF)

Oocytes and early embryo in vitro culture (IVC) was a key technology of mammalian embryo engineering and was the basis of biotechnology such as genetically modified (GM) and cloning research.The factors,which affect the efficiency of in vitro culture for oocytes and early embryo,include the composition,ion concentration and osmotic pressure of the culture medium,and the gas composition and temperature of the culture environment. Recent studies have shown that the oxygen tension in the culture systems can significantly affect the development of embryos,and this effect is mainly regulated by the hypoxia-inducible factor (HIF). This paper reviewed the effects of the oxygen tension on oocyte maturation and early embryo development,and the structure and regulation mechanism of HIF were discussed,in order to provide a theoretical basis for further explore the application of hypoxia method in the in vitro culture of oocytes and early embryo.     

Effects of different diets on viability, and gut morphology and bacteriology in captive willow ptarmigan chicks (Lagopus L. lagopus)

Young willow ptarmigan chicks were fed an artificial chick diet (ACD), and different supplements. One group got ascorbic acid supplements, another neomycin and Oxytetracyclin in the water, and a third was given ascorbic acid and sprouts of Vaccinium myrtillus. Chicks that were fed ACD only, showed low weight gain and high mortality rate. The numbers of bacteria in the gut were high and the incidence of villous epithelium erosion and bacteria colonizing the gut epithelium were high.Admixture of ascorbic acid to the diet improved the viability of the chicks and reduced the number of coliforms in the gut.Antibiotics in the water reduced the number of C. perfringens, enterococci and coliforms in the gut, and improved the gut epithelium conditions.Combined supplements of ascorbic acid and Vaccinium myrtillus resulted in strongly viable chicks with healthy gut epithelium. C. perfringens was not demonstrated in the gut, and the numbers of enterococci and coliforms were low.     

The effect of bpV(HOpic) on in vitro activation of primordial follicles in cultured swine ovarian cortical strips

The vanadate‐derivative dipotassium bisperoxo (5‐hydroxy‐pyridine‐2‐carboxylic) oxovanadate (V) (bpV(HOpic)), a pharmacological inhibitor of phosphatase and tensin homolog (PTEN), has been used in ovarian follicle culture systems for activation of follicular growth in vitro and suggested to be responsible for primordial follicle survival through indirect Akt activation. For pig ovarian tissue, it is still not clear which culture medium needs to be used, as well as which factors and hormones could influence follicular development; this also applies to bpV(HOpic) exposure. Therefore, ovarian cortical strips from pigs were cultured in 1 µM bpV(HOpic) (N = 24) or control medium (N = 24) for 48 hr. Media were then replaced with control medium and all tissue pieces incubated for additional 4 days. The strips were embedded in paraffin for histological determination of follicle proportions at the end of the culture period and compared to histological sections from tissue pieces without cultivation, which had been embedded right after preparation; comparison of healthy follicles for each developmental stage was performed to quantify follicle survival and activation. After 6‐day culture, follicle activation occurred in tissue samples from both cultured groups but significantly more follicles showed progression of follicular development in the presence of 1 µM bpV(HOpic). The amount of non‐vital follicles was not significantly increased during cultivation. BpV(HOpic) affects pig ovarian follicle development by promoting the initiation of follicle growth and development, similar as in rodent species and humans.     

Oocyte holding in the Iberian red deer (Cervus elaphus hispanicus): Effect of initial oocyte quality and epidermal growth factor addition on in vitro maturation

Current in vitro embryo production protocols in the Iberian red deer (Cervus elaphus hispanicus) need to be optimized; oocyte harvesting in situ followed by overnight holding could reduce the human effort and shipping costs. In our work, post‐mortem ovaries were retrieved, and the oocytes were harvested and allocated to G1 group (good quality) or G2 + G3 group (low quality). The oocytes were separately subjected to immediate in vitro maturation (IVM) or held overnight in a holding medium composed of 40% of TCM 199 with Earle's salts, 40% TCM 199 with Hanks' salts and 20% fetal bovine serum (FBS), at room temperature (16 hr). In vitro maturation was carried out in a basal medium supplemented or not with 50 ng/ml of epidermal growth factor (EGF). Our data showed that addition of EGF to the maturation medium increases the percentage of G1 oocytes reaching metaphase II (3.9% vs. 50%, basal vs. EGF; p < .001) and decreased their degeneration rate (69.9% vs. 22.2%, basal vs. EGF; p < .01) when oocytes were immediately matured. Overnight holding increased the meiotic competence of G1 oocytes (37.5% matured in basal medium) and EGF increased prophase arrest in G2 + G3 oocytes (16.1% vs. 38.8% in germinal vesicle [GV] stage in basal medium vs. EGF added medium; p < .05). Our data demonstrate that oocyte holding can be used in Iberian red deer oocytes. Interestingly, EGF addition increases the oocytes' meiotic competence in immediately matured oocytes but not after oocyte holding depending upon initial oocyte quality.     

Ontogenetic dietary shift and morphological correlates for Diplodus capensis (Teleostei: Sparidae) in southern Angola

Allometric growth patterns of functional morphological variables that reflect resource use in fish were correlated with the diet of Diplodus capensis in southern Angola. A total of 114 individuals (76–336 mm FL) were collected for stomach content analysis and a further 35 individuals (80–320 mm FL) for morphometric analysis. The major dietary components belonged to the groups Chlorophyta, Cirripedia, Rhodophyta and Bivalvia, and made up 71.7% of the diet in terms of a computed ranking index. There was a clear ontogenetic dietary shift with small fish feeding predominantly on algae and large fish on barnacles and mussels. This increasingly durophagous diet corresponded with a positive allometric growth pattern in the gape dimensions (width, height and surface area), adductor mandibulae muscle process and incisor width. Conversely, the relative gut length followed an isometric growth pattern, suggesting a decreased absorptive capacity. This isometric growth pattern is attributed to the better digestibility of the durophagous prey items when compared with algae and further corroborates a link between the morphology and an ontogenetic dietary shift observed in this species.     

The diet,food selectivity and niche of Rhabdosargus sarba (Teleostei: Sparidae) in Natal estuaries

The juveniles and subadults of Rhabdosargus sarba are one of the most common components of the benthic fish community of Natal estuaries. The diet consists mainly of aquatic macrophytes and filamentous algae although a significant proportion is made up of bivalves and the amphipod Grandidierella lignorum. The last two taxa are positively selected when little plant material is consumed. R. sarba feeds during daylight with a peak in the early afternoon. The relationship between R. sarba and the very similar sympatric R. holubi and R. auriventris are discussed in relation to their diet, divergence and distribution. Resource partitioning in Natal estuaries between R. sarba, the Gerreidae and Pomadasys commersonii is also discussed.     

NaCl胁迫对苜蓿幼苗生长及不同器官中盐离子分布的影响

研究了不同浓度NaCl胁迫下(盐浓度处理为0,0.1%,0.3%,0.5%,0.7%,0.9%)苜蓿不同器官的生长情况(存活率、根长、茎长、叶片数),以及不同器官中Na⁺、K⁺、Ca²⁺、Mg²⁺的含量,并分析了K/Na、Ca/Na、Mg/Na值的变化。结果表明,存活率、根长、茎长与盐浓度呈负相关,且当NaCl≤0.1%时,有利于苜蓿的生长。除根茎叶中Na⁺含量、根中Ca²⁺与盐浓度呈正相关,茎叶中的Ca²⁺与盐浓度相关性不明显外,根茎叶中K⁺、Mg²⁺、K/Na、Ca/Na、Mg/Na均随盐浓度增加而显著下降,且盐处理与对照差异显著(P＜0.05)。另外从离子角度讨论了盐对苜蓿的毒害以及耐盐的原因。     

Rumen microbial variation and nutrient utilisation in mithun (Bos frontalis) under different feeding regimes

The aim of the study was to investigate the effect of feeding different diets on fermentation, enzyme activities and microbial population in the rumen fluid of mithun (Bos frontalis). In a randomized block design, 20 male mithun (6–8 months of age, 152 ± 12.6 kg body weight) were randomly divided into four experimental groups (n = 5/group) and fed experimental diets ad libitum for 180 days. The diet R₁ contained tree foliages (TF), R₂ comprised of 50% concentrate mixture (CM) and 50% TF, R₃ contained 50% CM and 50% rice straw, and R₄ contained 50% CM, 25% TF and 25% rice straw. Rumen liquor was collected at 0 and 180 days of the experiment for estimation of different ruminal parameters and a digestion trial was conducted at the end of the experiment. Rumen fluid was analysed for pH, ammonia nitrogen (NH₃‐N), total‐N, ruminal enzymes, short chain fatty acid (SCFA) and microbial profile. The relative quantification of ruminal microbes was carried out with real‐time PCR using bacteria as the house keeping gene. The dry matter intake, nutrients digestibility, body weight gain, NH₃‐N, total‐N, carboxymethyl cellulase, avicelase, xylanase, amylase, protease and molar proportion of butyrate were (p < 0.05) higher in mithun fed R₂, R₃ and R₄ compared to those fed R₁ diet. In contrast, increased (p < 0.05) ruminal pH, molar proportion of acetate and, acetate to propionate ratio was recorded in mithun fed only TF than those fed concentrate supplemented diets. Similarly, an increase (p < 0.05) in the population of Fibrobacter succinogenes, Ruminococcus flavefaciens and total bacteria were evident in mithun fed R₂, R₃ and R₄ compared to those fed R₁. Therefore, it is concluded that TF 25% and/or rice straw 25% along with CM 50% may be fed to the growing mithun for improved rumen ecology, nutrient utilization and thus better performance under stall fed system.     

不同水分环境下玉米叶面积QTL定位及候选基因分析

玉米叶面积的大小及分布特征不仅影响其光合效率、蒸腾速率,而且与其耐旱性、耐密性、抗倒伏性及产量形成紧密相关。深入剖析不同水旱环境下玉米不同生育时期不同叶位叶面积的分子遗传机理对玉米耐旱高产新品种的选育具有重要意义。本研究以构建的2套F_2∶3群体为试材,在8种水分环境下,采用复合区间作图法（CIM）和基于混合线性模型的复合区间作图法（MCIM）对玉米相应叶（V₁₈时期第10片叶、R₁时期穗三叶）叶面积进行单环境和多环境联合QTL分析;参考玉米基因组B73 RefGen_v3挖掘稳定表达的QTLs （sQTLs）区间内的候选基因,并对其进行功能分析。结果表明,采用CIM法,单环境下2个生育时期2套F_2∶3群体间总共定位到了7个玉米相应叶叶面积QTLs,主要受显性（81.0%）、部分显性（14.3%）和超显性（4.7%）等遗传效应的调控,其中在干旱环境下定位到了5个QTLs。采用MCIM法,在2套F_2∶3群体间总共检测到6个相应叶叶面积的联合QTLs,其中1个表现为显著的QTL与环境的互作（QTL×E, Bin 2.08~2.09）,1对QTLs （Bin 1.08~1.10与 Bin 2.08~2.09）参与了显著的加性与加性（AA）上位性互作。结合CIM和MCIM法进一步分析在2套F_2∶3群体间检测到了6个sQTLs,其分别位于Bin 1.08~1.10、Bin 2.08~2.09、Bin 4.08~4.09、Bin 6.05、Bin 8.03和Bin 10.03处,并在这些sQTLs区间内确定了12个玉米叶发育相关候选基因。采用生物信息学,总共收集了75个玉米叶发育相关候选基因,通过系统进化树分析表明,这些候选基因划分为3大进化分支,且上述检测到的12个候选基因分布于这3大进化分支上。这些结果为系统地解析玉米不同生育时期不同水旱环境下相应叶叶面积的分子遗传机理提供理论依据,检测到的sQTLs可作为叶面积改良的重要染色体区段,检测到的候选基因为其进一步克隆、功能分析及育种应用提供了信息参考。     

盐胁迫下外源Si对甘草种子萌发及幼苗K~+、Na~+含量的影响

以乌拉尔甘草(Glycyrrhiza uralensis)种子为试验材料,研究在NaCl(100和200mmol/L)胁迫下,不同浓度Si(0、0.4、0.8、1.2、1.6和2.0mmol/L)对种子萌发及幼苗根冠比和K~+、Na~+含量的影响。结果表明:外源Si对盐胁迫下种子的发芽率、发芽势和发芽指数均具有显著的促进作用,且Si浓度在0.4和2.0mmol/L下作用最佳;Si可显著增加幼苗的根冠比,幼苗可通过增加K~+吸收,减少Na~+吸收来维持根系内K~+、Na~+的平衡。     

Water intake in domestic rabbits (Oryctolagus cuniculus) from open dishes and nipple drinkers under different water and feeding regimes

Rabbits (Oryctolagus cuniculus) are often presented suffering from urolithiasis. A high water intake is important in the prophylaxis of uroliths. We investigated the influence factors for water intake using 12 rabbits subjected to different feed and water regimes with practical relevance: Hay, fresh parsley, a seed mix and two different pelleted feed were offered in diverse combinations. Water was provided either by open dish or nipple drinker. Water was accessible ad libitum except for four treatments with 6 h or 12 h water access. Under the different feeding regimes, the drinker had no influence on water intake, but faecal dry matter content was significantly higher with nipple drinkers [60.0 ± 2.1 vs. 57.2 ± 2.1% of wet weight (mean ± 95% confidence interval), p = 0.003]. Dry food led to a higher drinking water intake but total water intake was still lower than with addition of 'fresh' food. With restricted water access, rabbits exhibited a significantly higher water intake with open dishes compared with nipple drinkers (54.9 ± 9.8 vs. 48.1 ± 8.2 g/kg(0.75) /day (mean ± 95% confidence interval), p = 0.04). High proportions of fresh parsley or hay in the diet enhanced total water intake and urine output, and led to lower urinary dry matter content and lower urinary calcium concentrations. Restricted access to drinkers led to a decreased total daily water intake and increased dry matter content of urine and faeces. For optimal water provision and urolith prophylaxis, we recommend a diet with a high 'fresh food' proportion as well as additionally hay ad libitum with free water access, offered in an open bowl.     

Anatomy of the vertebral column,ribs and sternum in orange rumped agouti (Dasyprocta leporina Linnaeus, 1758): Structural and Functional perspectives

The axial skeleton of orange rumped agouti, Dasyprocta leporina, was studied for better understanding of its locomotor behaviour. The bones from eight adult agoutis of both sexes were observed for their anatomical features and functional significance. The vertebral formula was found to be C₇T₁₂L₇S₅Cy_5–6. The well-developed occipital crest, caudally oriented prominent axis spine and well-developed transverse processes from C₃–C₇ indicated a highly flexible neck with greater sagittal mobility. Articular facets were horizontal in anterior series while oblique in the posterior series, which enabled them to perform both lateral and sagittal movements during locomotion. The caudally directed thoracic spines, T₁₂ as anticlinal vertebra and prominent mamillary process in the posterior series were suggestive of strong dorso-ventral flexion/extension and rotation. The robust lumbar vertebrae, well-developed transverse processes with cranio-ventral extension, were the feature for powerful sagittal/dorsoventral movement. The presence of spinous processes and well-developed transverse processes in all caudal vertebrae was an indication of a highly movable tail. The ribs were 13 pairs with first seven as sternal and six as asternal. They were laterally compressed in the anterior series as a cursorial adaptation. A strong muscular attachment to vertebrae provides this rodent speed, agility, dexterity and strength suitable for survival in food chain.     

Morphological characteristics of the forebrain in the donkey (Equus asinus): A compared atlas of magnetic resonance imaging and cross-sectional anatomy

The brain is the most essential part of the central nervous system which regulates and coordinates all body activities. Based on its phylogenetic development from the neural tube, the brain is divided into rhombencephalon (hindbrain), mesencephalon (midbrain) and prosencephalon (forebrain). The present study is achieved to describe the morphological characteristics of the normal forebrain in the donkey using the matched magnetic resonance imaging (MRI) and cross-sectional anatomy. Ten cadaveric heads of healthy adult donkeys of both sexes were used. Two heads were examined using a 1.5 Tesla MRI scanner, and the brains of the other heads were gently extracted; six brains were sectioned into transverse, dorsal and sagittal slices, and two brains were grossly inspected. MR images were selected in correlation to their closely corresponding gross sections. Both cross-sectional anatomy and MRI scans showed extensive gyration of the neocortex. The forebrain structures appeared with variable intensities on three sequences, Flair, T1-weighted and T2-weighted MRI, enabling comprehensive evaluation of the relevant neuroanatomical structures. The present study provided a precise neuroanatomical atlas of the forebrain in the donkey which could help in the quick and efficient interpretation of clinical diseases of the forebrain, localization of the forebrain functions and evolutionary neurobiology.     

A new chelonibiid from the Miocene of Zanzibar (Eastern Africa) sheds light on the evolution of shell architecture in turtle and whale barnacles (Cirripedia: Coronuloidea)

The fossil history of turtle and whale barnacles (Coronuloidea: Chelonibiidae, Platylepadidae, Coronulidae and †Emersoniidae) is fragmentary and has only been investigated in part. Morphological inferences and molecular phylogenetic analyses on extant specimens suggest that the roots of whale barnacles (Coronulidae) are to be found among the chelonibiid turtle barnacles, but the hard-part modifications that enabled early coronuloids to attach to the cetacean skin are still largely to be perceived. Here, we reappraise a fossil chelonibiid specimen from the Miocene of insular Tanzania that was previously referred to the living species Chelonibia caretta. This largely forgotten specimen is here described as the holotype of the new species †Chelonibia zanzibarensis. While similar to C. caretta, †C. zanzibarensis exhibits obvious external longitudinal parietal canals occurring in-between external longitudinal parietal septa that abut outwards to form T-shaped flanges, a character so far regarded as proper of the seemingly more derived Coronulidae and Platylepadidae. Along with these features, the presence of a substrate imprint on the shell exterior indicates that †C. zanzibarensis grasped its host's integument in much the same way as coronulids and platylepadids, albeit without the development of macroscopic parietal buttresses and bolsters. Thin section analyses of the inner parietal architecture of some extant and extinct coronuloids conclusively demonstrate that vestiges of comparable external parietal microstructures are present in some living members of Chelonibiidae. This observation strengthens the unity of Coronuloidea while significantly contributing to our understanding of the evolution of the coronuloid shell structure in adapting to a diverse spectrum of hosts.     

The effects of lactated Ringer's solution (LRS) or LRS and 6% hetastarch on the colloid osmotic pressure, total protein and osmolality in healthy horses under general anesthesia

ObjectiveTo investigate changes in colloid osmotic pressure (COP), total protein (TP) and osmolality (OSM) during anesthesia in horses given intravenous lactated Ringer’s solution (LRS) or LRS and hetastarch (HES).Study designProspective, clinical trial.AnimalsFourteen horses presented for surgery. Mean age 8.3 ± 1.9 years; mean weight 452 ± 25 kg.MethodsHorses were premedicated with xylazine intravenously (IV); anesthesia was induced with ketamine and diazepam IV, and maintained with sevoflurane. Butorphanol was administered IV with pre-medications or immediately after induction. Xylazine was administered IV for recovery if necessary. LRS was administered IV to all horses with a target rate of 5–10 mL kg^?1 hour^?1. Half of the horses also received 6% HES, 2.5 mL kg^?1 over 1 hour in addition to LRS. Horses that received LRS only were considered the LRS group. Horses that received both LRS and HES were considered the LRS/HES group. Blood was drawn pre- and post-anesthesia, immediately following induction, and every 30 minutes throughout anesthesia. COP, TP and OSM were measured.ResultsCOP and TP significantly decreased at similar rates for both treatment groups from pre-anesthetic values. Pre-anesthetic COP was significantly greater in the LRS group when compared to the LRS/HES group pre-, post- and throughout anesthesia. In the LRS group post-anesthetic OSM was significantly different than the pre-anesthesia value and that for the LRS/HES group.Conclusions and clinical relevanceAdministration of IV HES (2.5 mL kg^?1, over 1 hour) in combination with LRS does not attenuate the decrease in COP typically seen during anesthesia with crystalloid administration alone. Based on these results, administration of HES at this rate and total volume would not be expected to prevent fluid shifts into the interstitium through its effects on COP.     

柳枝稷和白羊草苗期水分利用与根冠比的比较

室内生长箱内柳枝稷和白羊草苗期水分利用与根冠比的试验结果表明，低水下柳枝稷光合速率降低，蒸腾速率增大，叶片水分利用效率减小，根长增加，整体水分利用率降低，但生产力的根冠分配与高水相差较小。低水也造成白羊草光合降低，但蒸腾速率减小幅度更大，叶片水分利用效率反而提高，主要原因是白羊草叶片气孔具有较强的随水分条件变化的调节作用。白羊草的根系在低水下伸长速率减慢，根长／株高降低，水分条件好时根系也能够迅速生长，特别是加强根系的生产力分配。不同水分下白羊草消耗单位水分形成的生产力均大于柳枝稷，说明白羊草在苗期对土壤水分变化的适应能力比柳枝稷要强。     

Morphological changes of physeal cartilage and secondary ossification centres in the developing femur of the house mouse (Mus musculus): A micro‐CT based study

In mammals, long bones are formed by ossification of a cartilaginous mould during early stages of development, through the formation of structures called the primary ossification centre, the secondary ossification centres (SOCs) and the physeal cartilages (PCs). The PC is responsible for long bone growth. The morphology of the PC and the SOCs varies during different stages of femoral growth. In this respect, several details involving the process of murine femoral development are lacking. In the present study, a morphological characterization of femur development from the embryonic period to adulthood in mice was studied using micro‐computed tomography (micro‐CT). To achieve this aim, femora were collected at embryonic day (E) 14.5, E16.5 and E18.5 and at postnatal day (P)1, P7, P14, P35, P46 and P52. CT images were obtained using a micro‐CT scanner (X‐SkyScan 1172; Micro Photonics) and analysed using the micro‐CT 3D visualization software Mimics (Materialise NV, Leuven, Belgium) and NRecon (Micro Photonics). The results of the present study revealed that the femur and its PCs and SOCs undergo morphological changes during different stages of development, including changes in their shape as well as position and thickness. These changes may be due to the response of the femur to mechanical loads imposed by muscle surrounding the bone during these stages of development. The result of the present study is important to improve our knowledge related to ossification and growth patterns of mouse femur during development.     

不同盐分条件下硅对两个高羊茅品种生物量分配和营养元素氮、磷、钾吸收利用的影响

频繁的灌溉,使人工草地的高羊茅生长在盐渍化或潜在盐渍化环境中。硅可以抑制植物对Na⁺的吸收,增加对N、P和K⁺的吸收,从而提高植物的耐盐性,且这种影响因植物种类和品种的不同而不同。本研究采用盆栽试验,研究了不同程度盐生境下施硅对两个高羊茅品种（抗性弱的K31和抗性强的XD）生物量和植株N、P、K⁺、Na⁺含量的影响。结果表明,随着盐浓度的增加,两个高羊茅品种地上和地下生物量、N、P、K⁺含量降低,Na⁺含量增加。施硅对地上和地下生物量、N、P、K⁺、Na⁺含量的影响与盐浓度有关,在中低盐浓度下,施硅增加了高羊茅的地上、地下生物量、根冠比、N、P、K⁺ 含量、K⁺/ Na⁺ 值,降低了Na⁺含量。高盐浓度时,施硅对高羊茅的生物量和N、P、K⁺、Na⁺含量没有影响。施硅对两个高羊茅品种地上和地下生物量、Na⁺、P含量的影响相似,对K⁺、N含量的影响不同,硅对耐盐性较强的XD地上部K⁺及N含量的有利影响优于耐盐性较弱的K31。结果表明,在低中盐浓度条件下,施硅可以促进高羊茅的生长,并且对耐盐性较强的高羊茅品种XD更为有利。