温度、光照和保存液对大鲵成熟精子存活状况的影响

本文对人工催产得到的大鲵精子,在不同温度、光照和保存液中的存活状况进行了观察研究。结果表明:15℃条件下体外保存大鲵精子原液,有光照时精子存活5h,无光照时存活10h;相同温度下精子原液经0.9%生理盐水、鱼用任氏液、蒸馏水、山泉水稀释后,精子在鱼用任氏液中存活时间最长,但与原液中保存精子的存活时间相比较有明显的缩短;通过对大鲵精子活力级别的判定,认为大鲵精子在水温20～25℃时受精能力最强。     

Skeletal development and deformities in cultured larval and juvenile seven-band grouper, Epinephelus septemfasciatus (Thunberg)

The seven‐band grouper, Epinephelus septemfasciatus (Thunberg), is currently recognized as a potential new species for aquaculture in Japan. This study describes normal and abnormal skeletal development of the jaw and vertebrae in cultured larvae and early juveniles of E. septemfasciatus. The ontogenetic stages at which skeletal deformities of jaw and vertebra developed were also described for this species. Osteological observations were made using a clearing and staining method for larvae and soft X‐ray photographs for juveniles. A high incidence of skeletal deformities was observed in the jaws and vertebral column during the larval and juvenile stages. Most of the jaw deformities were explained by an abnormal maxilla curvature. Jaw deformities were visually evident from flexion stage after ossification of the deformed elements. Deformities in the vertebral column (mostly lordosis) were observed from the post‐larval stage and became more evident as growth proceeded. The lordosis generally occurred on the positions of the 8–11th vertebra. These types of deformities are compared with those of other species, and possible causative factors of the skeletal deformities are discussed.
     

Histopathological studies on viral nervous necrosis of sevenband grouper, Epinephelus septemfasciatus Thunberg, at the grow-out stage

Abstract Viral nervous necrosis caused by sevenband grouper nervous necrosis virus (SGNNV) has occurred in grow-out stages (0-3 years old) of sevenband grouper, Epinephelus septemfasciatus, since the 1980s. In the present study, based on histopathological features of the central nervous system (CNS) in naturally diseased fish, pernasal infection experiments using grow-out fish were performed and pernasal infection was established as a putative invasion route of SGNNV. The definite SGNNV-targeted cells were determined by histopathological studies including indirect fluorescent antibody test and electron microscopy. Nerve cells in the olfactory lobe were most extensively necrotized with vacuolation followed by infiltration of microglia and macrophages. Purkinje cells and Golgi cells were extensively infected in the cerebellum. Megalocells and small nerve cell nuclei were also infected in the preoptic area, thalamus, medulla oblongata and spinal cord. Only a few small nerve cells were infected in the olfactory bulb and optic tectum. The retina of some diseased fish displayed vacuolated bipolar cells of the inner nuclear layer and in the ganglion cell layer. These SGNNV-infected nerve cells displayed viroplasmic inclusions containing virions, vacuoles and myelin-like structures. Based on observed histopathological changes, the lesion of the CNS was characterized by encephalitis but not encephalopathy.     

Histological observations of seasonal reproductive and lunar-related spawning cycles in the female honeycomb grouper Epinephelus merra in Okinawan waters

Seasonal reproductive and lunar-related spawning cycles of the female honeycomb grouper Epinephelus merra inhabiting Okinawan waters were examined by histological observation of ovaries. Gonadosomatic index (GSI) increased beginning in May and peaked in June. Histological observations revealed that many oocytes laden with yolk were present in the ovaries from May to August. From September to October, ovaries were occupied by immature oocytes. These results suggest that the reproductive season of E.   merra lasts for 4 months from May through August in Okinawan waters. When the fish were collected according to the lunar cycle, GSI increased with the approach of the full moon. Oocytes at various development stages were observed from the first quarter to the full moon. Fresh ovulatory follicles were present in the ovaries around the last quarter moon. These results suggest that E.   merra has a lunar spawning cycle and spawns between the full moon and the last quarter moon. Ovarian features of the fish collected around the last quarter moon were different among individuals; some fish had many oocytes at the tertiary yolk stage in the ovaries, while the ovaries of the others were occupied by the oocytes at the peri-nucleolus and the oil droplet stages. This observation suggests that a minor release of eggs occurs in this species before or after a major spawning lunar day. Oocytes at the migratory nucleus and the maturation stages were not observed in any ovarian samples. This may mean that maturation of oocytes is related to the spawning behavior of this species and makes rapid progress in the process of aggregation at the spawning sites.     

Genetic diversity of wild mekong giant catfish Pangasianodon gigas collected from Thailand and Cambodia

The Mekong giant catfish Pangasianodon gigas is endemic to the Mekong River and is a critically endangered species. The genotypes of the microsatellite DNA (msDNA) and mitochondrial DNA (mtDNA) markers (right domain of the control region) were detected to evaluate the present status of genetic divergence of this species from the Mekong River in Thailand and Cambodia. The observed and expected heterozygosity values of Mekong giant catfish in Thailand and Cambodia were relatively low in comparison with those of other nonendangered freshwater fish species. These two populations from Thailand and Cambodia showed similar levels of genetic diversity, as evaluated by the 384 nucleotides of the mtDNA control region with 13 haplotypes. The pairwise F _ST value between the two populations based on the genotype frequencies of msDNA and mtDNA markers suggested a close genetic relationship between the populations in Thailand and Cambodia. The results of this study support the conclusion that the Mekong giant catfish is critically endangered. Care should be taken to sustain the genetic diversity of this species, as the level of genetic variability has already decreased in the wild population. This species is a target species for an ongoing stock enhancement program in the Mekong River in Thailand. It is proposed to apply these markers for proper broodstock management, such as for minimal kinship selective breeding in the hatchery.     

不同种植年限皇竹草产量与生物学性状关系分析

摘要：为研究不同种植年限皇竹草（Pennisetum hydridum）产量与生物学性状之间的关系,对生长1~3年的皇竹草一年内不同时期的产量及生物指标进行跟踪观测。结果表明,皇竹草植株株高、分蘖数、有效茎数和产量等均随种植年限的增加而明显提高,其相互间关系极显著正相关。 通过相关性分析和通径分析发现,有效茎数、节数和分蘖数对皇竹草产量的直接和间接作用最大。 同时,一元回归分析显示,以节数为变量的拟合程度最高,而五元线性回归模型Y=-23.742+0.031X1+0.261X2+0.187X3+0.025X4+0.825X5,R2=0.822（P<0.01）对不同种植年限的田间实际产量拟合最佳。综合分析认为,植株分蘖数、有效茎数和节数为皇竹草产量的最重要影响指标。     

日本血吸虫幼虫巨大致死基因片段的克隆、表达及免疫效果分析

为研究日本血吸虫(Schistosoma japonnicum)幼虫巨大致死基因(Lethal giant larvae)的生物学功能,本研究应用荧光定量RT-PCR分析SjLgL基因在S.japonnicum不同发育阶段的表达情况,构建重组表达质粒pET-SjLgL进行诱导表达,对重组蛋白进行western blot鉴定;重组蛋白刺激免疫动物进行免疫保护试验,检测IL-4、IL-10、TNF-α和IFN-γ的变化.结果表明,SjLgL基因在S.japonnicum不同发育时期均有表达,成虫期略高于童虫,雄虫高于雌虫.Western blot鉴定结果显示,该重组蛋白具有良好的抗原性.免疫保护试验结果表明,免疫小鼠分别获得了22.34％的减虫率和55.49％的肝脏减卵率.血清学检测结果显示重组蛋白免疫小鼠后产生了特异性IgG抗体.细胞因子检测结果表明,重组蛋白刺激免疫动物主要诱发产生Th1型的免疫应答.本实验结果表明该重组蛋白在小鼠体内可诱导产生部分免疫保护效果.     

“石棉巨型”核桃主要经济性状研究

在资源调查的基础上,针对“石棉巨型”核桃及“川早1号”、“川早2号”、“川早3号”、“双早”、“蜀玲”核桃的腹径、缝径、果高、壳厚、单果重、仁重、出仁率、粗脂肪含量、粗蛋白含量9个主要经济性状指标,采用相关性分析、主成分分析及聚类分析等方法,对绵竹县核桃资源进行综合评判.结果表明:“石棉巨型”核桃坚果大小明显高于其它5个杂交早实核桃良种,表现出巨型果的优势,在出仁率、粗脂肪含量等方面表现良好;壳厚、单果重、仁重两两呈极显著正相关.主成分分析结果表明,6个核桃样本主成分综合得分依次为:“石棉巨型”＞“川早3号”＞“川早2号”＞“川早1号”＞“双早”＞“蜀玲”.聚类分析得出,“石棉巨型”核桃单独聚为一类,其它5个品种聚为另一类,表明巨型核桃与其它5个品种表现出较远的亲缘关系.     

Possible application of mibolerone for induced sex inversion of grouper Epinephelus coioides

SUMMARY: Thirty immature juvenile grouper Epinephelus coioides (19–168 g bodyweight, BW) were randomly stocked in four units 6 t tanks to determine if mibolerone can be used to induce sex inversion in groupers. After acclimatization and weaning to artificial feed, the feed given daily (4% BW/day) was supplemented with 0, 50, 100, and 200 μg mibolerone/kg feed for about 18 weeks. Thereafter, the hormone treatment was withdrawn and the experiment was terminated at Week 24. Ten fish were killed for gonad histology at stocking to serve as an initial control while about three to five fish were killed every 8 weeks. In general, ovaries of initial controls showed the presence of moderate stromal cells and gonia and few primary oocytes. At Weeks 8 and 16, ovaries of the control fish (0 μg/kg) were similar to that of the initial control except that primary oocytes increased at Week 24. Gonads of fish fed diets containing 100 and 200 μg/kg had none to moderate spermatocytes and few spermatids at Week 8 and 16, although spermatozoa were not observed, indicating that the fish were undergoing spermatogenesis. Spermatogenesis at 50 μg/kg was not as advanced since only few spermatocytes occurred at Weeks 8 followed by moderate gonia and no spermatocytes and spermatids at Week 16. However, the presence of few primary oocytes was observed when mibolerone was withdrawn suggesting that sex-inversed fish reverted back to a female condition. These results show that sex inversion in juvenile grouper can be induced by oral administration of mibolerone and may have possible application on mature females to produce functional males.     

Adjuvant effects of recombinant giant panda (Ailuropoda melanoleuca) IL-18 on the canine distemper disease vaccine in mice

Canine distemper virus (CDV) is a morbillivirus known to cause morbidity and mortality in a broad range of animals. Giant pandas (Ailuropoda melanoleuca), especially captive ones, are susceptible to natural infection with CDV. Interleukin-18 (IL-18) is a powerful adjuvant molecule that can enhance the development of antigen-specific immunity and vaccine efficacy. In this study, a giant panda IL-18 gene eukaryotic expression plasmid (pcAmIL-18) was constructed. Female BALB/c mice were muscularly inoculated with the plasmids pcAmIL-18, pcDNA3.1 and PBS, respectively. They were subsequently injected with an attenuated CDV vaccine for dogs, and the induced humoral and cellular responses were evaluated. The results showed that pcAmIL-18 remarkably improved the level of specific antibody, IFN-γ and IL-2 in mice sera, the T lymphocyte proliferation index and the percentage of CD4⁺ and CD8⁺ cells. These data indicated that pcAmIL-18 is a potential adjuvant that promotes specific immunity.