Polymorphism and heterogeneity of transferrins in some species of the fish family Cyprinidae

In fish of the family Cyprinidae extensive polymorphism and heterogeneity of serum transferrins were found. Ten variants of transferrins were detected in barbel, 9 in dace and bleak, 7 in chub and bream, 6 in rudd, 5 in roach, white bream, silver carp, big head and crucian carp, 3 in tench, and 2 variants in nase, ide and goldfish. All the variants observed bound radioactive iron. The phenotype patterns indicated a simple genetic determination, by a set of codominant alleles at one locus, and this even in tetraploid species. Transferrins of some species were partly isolated. The transferrins of all 22 species analysed were heterogeneous and homozygous pheno-types had 1 to 6 zones of various intensities, binding ⁵⁹Fe. In barbel both single-zone and two-zone variants were present and transferrins of roach had variable heterogeneity. Sialic acid was found only in transferrins of silver carp, big head and in the two-zone variants of barbel. The occurrence of phenotypes with single and double zones of transferrins in barbel indicates the possibility of genetic determination of the inability of single-zone variants to bind sialic acid.     

Age-dependent changes in in vivo ethanol metabolism and in the activity of hepatic enzymes involved in ethanol oxidation and microsomal functions

The study of the influence of the age of the animals (13 to 53 weeks) on the rate of ethanol metabolism in vivo and the total activity of liver alcohol dehydrogenase and microsomal ethanol oxidizing system showed a progressive decline with age. These effects were observed concomitantly with a diminution in the content of cytochrome P-450 and microsomal functions related to oxidative and free-radical mediated reactions, namely, NADPH oxidase activity, NADPH-dependent oxygen uptake and NADPH-or t-butyl hydroperoxide-induced chemiluminescence. It is concluded that ageing is accompanied by a diminution in the total oxidative activity of the liver tissue, which would explain the depression in basal and ethanol-induced lipid peroxidation found in the oldest group of rats studied.     

松墨天牛成虫标本保存及其DNA提取质量比较

【目的】探讨适合DNA提取的天牛成虫标本保存方法。【方法】采用SDS-蛋白酶K消化法对液氮中冷冻保存、无水乙醇-20℃冷冻保存、无水乙醇室温保存和干标本室温保存且保存时间在2年以上的松墨天牛Monochamus alternates Hope成虫标本基因组DNA进行提取,并对不同保存方式提取的DNA样本进行了质量比较和分析。【结果】在上述常见的松墨天牛成虫标本4种保存方式中,以液氮中冷冻保存效果最佳,其次为无水乙醇-20℃冷冻保存,插针干标本室温保藏效果最差。利用昆虫线粒体基因CO I和CO II的通用引物从上述DNA中均能够成功扩增出目的片段,测序结果证实扩增片段符合预期。【结论】液氮和无水乙醇-20℃冷冻保存适合松墨天牛成虫标本长期保存,且不影响后续的PCR扩增和测序。     

Polymorphism of A, B and C loci of lactate dehydrogenase in European fish species of the Cyprinidae family

In 24 fish species of the Cyprinidae family, belonging to 21 genera, isoenzyme patterns of lactate dehydrogenase (LDH) were determined, which could be classified in the majority of cases into 3 main groups. Isoenzyme patterns in natural hybrids of roach and rudd, roach and bream, roach and bleak were also analysed. In bitterling, polymorphism was observed in B locus of LDH. In white bream polymorphism exists in the A locus. In bream, rudd, silver carp and barbel polymorphism was found in C loci. Isoenzyme patterns indicate that in each case the polymorphism is genetically controlled by two alleles at a single locus. The populations investigated were in Hardy-Weinberg equilibrium. No significant differences were found in the activity of liver LDH in various polymorphic types of C loci of bream and rudd.     

An alternative method for the preservation of tropical fish larvae

Preservation of newly hatched fish larvae in a 30% ethanol freshwater solution kept at −19° C allowed accurate measurement of morphology and otolith dimensions with minimal error due to shrinkage compared to other concentrations of ethanol or gluteraldehyde.     

五种淡水鱼类幼鱼游泳能力的比较

为了探讨栖息于不同生境中鱼类的游泳能力和偏好游泳速度及其生理机制,本研究以中华倒刺鲃(Spinibarbus sinensis)、异育银鲫(Carassius auratus gibelio)、岩原鲤(Procypris rabaudi)、青鱼(Mylopharyngodon piceus)和胭脂鱼(Myxocryprinus asiaticus) 5种鱼的幼鱼为对象,在(25±1)℃条件下测定了5种鱼类的标准代谢率(SMR)、最大代谢率(MMR)、有氧代谢范围(MS)、临界游泳速度(U_crit)、最大匀加速游泳速度(U_cat)和偏好游泳速度(U_pref)。结果发现:5种实验鱼中,中华倒刺鲃的游泳能力最强,游泳能力较差的为青鱼和胭脂鱼; 5种鱼之间的代谢和游泳能力差异显著,其偏好游泳速度主要集中在(10～24.5cm·s^-1)区域。研究表明,鱼类游泳能力的种间差异可能主要由心鳃系统相关的呼吸能力和体型相关的游泳效率所决定。本研究提供的有关鱼类游泳能力、偏好游泳速度等资料对于鱼道设计等有一定的参考价值...     

Sequence capture phylogenomics of historical ethanol‐preserved museum specimens: Unlocking the rest of the vault

Natural history collections play a crucial role in biodiversity research, and museum specimens are increasingly being incorporated into modern genetics‐based studies. Sequence capture methods have proven incredibly useful for phylogenomics, providing the additional ability to sequence historical museum specimens with highly degraded DNA, which until recently have been deemed less valuable for genetic work. The successful sequencing of ultraconserved elements (UCEs) from historical museum specimens has been demonstrated on multiple tissue types including dried bird skins, formalin‐fixed squamates and pinned insects. However, no study has thoroughly demonstrated this approach for historical ethanol‐preserved museum specimens. Alongside sequencing of “fresh” specimens preserved in >95% ethanol and stored at ?80°C, we used extraction techniques specifically designed for degraded DNA coupled with sequence capture protocols to sequence UCEs from historical museum specimens preserved in 70%–80% ethanol and stored at room temperature, the standard for such ethanol‐preserved museum collections. Across 35 fresh and 15 historical museum samples of the arachnid order Opiliones, an average of 345 UCE loci were included in phylogenomic matrices, with museum samples ranging from six to 495 loci. We successfully demonstrate the inclusion of historical ethanol‐preserved museum specimens in modern sequence capture phylogenomic studies, show a high frequency of variant bases at the species and population levels, and from off‐target reads successfully recover multiple loci traditionally sequenced in multilocus studies including mitochondrial loci and nuclear rRNA loci. The methods detailed in this study will allow researchers to potentially acquire genetic data from millions of ethanol‐preserved museum specimens held in collections worldwide.     

Modelling temperature-driven changes in species associations across freshwater communities

Due to global climate change–induced shifts in species distributions, estimating changes in community composition through the use of Species Distribution Models has become a key management tool. Being able to determine how species associations change along environmental gradients is likely to be pivotal in exploring the magnitude of future changes in species’ distributions. This is particularly important in connectivity-limited ecosystems, such as freshwater ecosystems, where increased human translocation is creating species associations over previously unseen environmental gradients. Here, we use a large-scale presence–absence dataset of freshwater fish from lakes across the Fennoscandian region in a Joint Species Distribution Model, to measure the effect of temperature on species associations. We identified a trend of negative associations between species tolerant of cold waters and those tolerant of warmer waters, as well as positive associations between several more warm-tolerant species, with these associations often shifting depending on local temperatures. Our results confirm that freshwater ecosystems can expect to see a large-scale shift towards communities dominated by more warm-tolerant species. While there remains much work to be done to predict exactly where and when local extinctions may take place, the model implemented provides a starting-point for the exploration of climate-driven community trends. This approach is especially informative in regards to determining which species associations are most central in shaping future community composition, and which areas are most vulnerable to local extinctions.     

Diagnostic PCR to identify five rare species of Cypriniformes in China

The major problem that fisheries managers face when trying to enforce the law is identifying fish species. This problem is even worse when it comes to identifying eggs, larvae, bloodstains, scales, mucous stains, and mixtures of these in markets, restaurants and fishing areas. To assist in the acquisition of urgently needed conservation and management data on rare and endangered fish catch and sale, we have developed and tested a highly streamlined molecular genetic approach to identify Cypriniformes fish species. We used species-specific primers and species-specific ladders in a seven-primer multiplex polymerase chain reaction format based on DNA sequence differences among species in the nuclear ribosomal second internal transcribed spacer (ITS2) to discriminate simultaneously among five rare and endangered Cypriniformes species in China. This technique distinguished samples with 100% accuracy. This genetic approach will be useful for the assessment, management and conservation of fish.     

Human exposures and potential health hazard assessment of Hg and Pb in some major imported frozen fish species in Nigeria

The levels of mercury and lead in three major species of imported frozen fish—Atlantic mackerel (Scomber scombrus), sardine (Sardinella sindesis), and scumbera (Scumbria spp)—were determined and potential health hazard resulting from their ingestion by Nigerians was assessed. Sample digestion was carried out as prescribed in the Food and Agricultural Organisation/Swedish International Development Agency wet digestion method. Quantitation of the Hg and Pb content of the digested samples and blanks was carried out spectrophotometrically using thiozone as the ligand. The ranges of level of Hg in the samples in mg/Kg were: 0.03–0.42 for Scomber scombrus and 0.03–0.37 and 0.020–0.31 for Scumbria and Sardinella sindesis, respectively. Also, Pb had value ranges in mg/Kg of 0.20–0.28, 0.24–0.32, and 0.15–0.32 in Scomber scombrus, Scumbria, and Sardinella sindesis, respectively. Potential health hazard assessment that was carried out showed no risk status for children and adults consuming Sardinella sindesis and scumbera and adults consuming Scomber scombrus (HQ < 1). The results further showed a risk (HQ = 1.022 and THI = 1.036) for children taking Scomber scombrus.     

五种丛生状棕榈植物叶热值的月变化研究

对5种丛生状棕榈植物一欧洲棕、细叶棕竹、小箬棕、玲珑椰子、缨络椰子的叶片热值和灰分含量的月变化进行了研究,结果表明,5种植物具有较高的灰分含量,但灰分含量的月变化趋势各不相同;欧洲棕、细叶棕竹、小箬棕3种植物干重热值的月变化趋势有些差异,同属的缨络椰子、玲珑椰子干重热值的月变化趋势基本相似;缨络椰子、小箬棕的干重热值与灰分含量具有显著的线性相关(P<0．05),而其余3种植物的干重热值与灰分含量相关性不显著(P>0．05);从去灰分热值的月变化来看,细叶棕竹与小箬棕,缨络椰子与玲珑椰子的月变化趋势相似。     

Metabolic changes during rooting in stem cuttings of five mangrove species

Vegetative propagation through rooting in stem cuttings in five tree mangroves namely Bruguiera parviflora, Cynometra iripa, Excoecaria agallocha, Heritiera fomes, and Thespesia populnea using IAA, IBA and NAA was reported. Spectacular increase in the root number was noted in the cuttings of H. fomes and C. iripa treated together with IBA (5000 ppm) and NAA (2500 ppm). The highest number of roots was obtained with IBA (2500 ppm) and NAA (500 ppm) in E. agallocha. B. parviflora and T. populnea responded better to IAA and IBA treatment. The species specific variation in the rooting response to exogenous application of auxins was reflected in the metabolic changes during initiation and development of roots in cuttings. Biochemical analysis showed increase of reducing sugar in the above-girdled tissues at initiation as well as subsequent development of roots which was further enhanced by the use of auxins. Decreases in the total sugar, total carbohydrate and polyphenols and increase in total nitrogen were recorded in the girdled tissues and the high C/N ratio at the initial stage helped in initiation of roots in all the species. Interaction of IBA and NAA promoted starch hydrolysis better than IAA and IBA during root development and subsequently reduced the C/N ratio and increased the protein-nitrogen activity during root development which suggest the auxin influenced mobilization of nitrogen to the rooting zone.Abbreviations IAA Indole-3-acetic acid - IBA Indole-butyric acid - NAA A-naphthalene acetic acid     

Maturation and hatching of eggs from silkworm ovaries preserved in liquid nitrogen

Ovarian imaginal discs prepared from fifth-instar larvae of the silkworm, Bombyx mori were treated with graded concentrations of glycerol, cooled at a rate of 1°C/min to ?35°C and preserved in liquid nitrogen for 2 days or more and then rapidly thawed (500°C/min). The frozen and thawed ovaries were transplanted into fifth-instar female larvae, in which more than 20% of the ovaries developed to produce mature eggs with a chorion according to the state of host development. By parthenogenetic activation, the mature eggs started embryogenesis and hatched to produce larvae. About 50% hatching occurred in the eggs developed in a C 108 × Cambodge host, and about 10% in a C 108 × Aojuku host. The hatched larvae completed post-embryonic development as did the normal larvae.     

Morphology and fertilizability of frozen human oocytes

Human oocytes were frozen and thawed by four methods previously used for cryopreser-vation of human embryos. Most of these oocytes were inseminated after thawing to assess their capacity to fertilize and form pronuclear ova. Their morphology was assessed by phase-contrast microscopy used in routine IVF. Twenty-three oocytes were examined by electron microscopy to critically evaluate the effects of cooling and cryopreservation and to confirm fertilization. Morphological survival was observed in more than 60% of the oocytes examined after freeze-thawing. The main features of cryoinjury were cracks in the zona pellucida, disruption of the plasma membrane and extensive disorganization of the ooplasm. Subtle changes in the cytosol of cumulus cells was also observed. Cooling to 0°C or ?6°C had little effect on cytoplasmic structure. Spindles were damaged in two frozen oocytes. Cumulus cell activity, sperm binding to the zona, sperm penetration of the zona seem to be largely unaffected by freeze-thawing. Fertilization was observed in eight oocytes after postthaw insemination and three embryos (8-cell to morula stages) were developed from pronuclear ova on further culture. Both monospermic and polyspermic fertilization were confirmed by electron microscopy and micronuclei were detected in three pronuclear ova. The genetic implications of these nuclear aberrations are discussed. These preliminary studies indicate that oocyte freezing needs to be integrated cautiously with clinical IVF by further assessment of embryos developed from frozen oocytes.     

Microsatellites from the vairone Leuciscus souffia (Pisces: Cyprinidae) and their application to closely related species

Eleven microsatellites were isolated from the vairone Leuciscus souffia (Risso 1826), an endangered fish that inhabits river systems in and around the Alps in Europe. The level of genetic diversity was assessed in 29 individuals of the subspecies L. s. souffia, and their variability was further estimated in seven individuals of a different subspecies, L. s. muticellus. Eight of these microsatellite loci were also applied to seven closely related cyprinid species. Availability of the reported microsatellite loci will facilitate the investigation of population genetic structure of these species with applications for the development of conservation strategies and phylogeographical approaches.     

Fruit-body production of test cultures ofFlammulina velutipes preserved for seven years by freezing at three different temperatures

Two strains ofFlammulina velutipes were cultured on PDA plates, and mycelial disks punched out using a cork borer were used for preservation. Five disks of a strain were put into a vial containing one of three cryoprotectants, 10% glycerol, 5% DMSO or 10% polyethylene glycol. Vials were then stored for 7 yr at −20°C, −85°C or liquid nitrogen temperature. The mycelial growth on PDA plates of the cryopreserved mycelial disks, as well as the usual subcultures, were tested two times. After the second test, spawns were prepared for fruit-body production tests by bottle cultivation from selected plates of the second growth tests. The yields of fruit-bodies varied among the cultures derived from the mycelial disks of the same strain preserved under different conditions. Variation in yields was observed even among the mycelial disks preserved at liquid nitrogen temperature, although the range of yield variation was narrower. The yield variation was obvious for the cultures which showed large retardation in the growth test. Four mycelial disks out of the six preserved at −20°C showed higher yields than those preserved at other temperatures. Among the cultures derived from strain FMC224, the control cultures preserved by subculture showed the lowest yield.