利用人皮肤模型评价杀菌类农药腐蚀/刺激性的研究

建立人皮肤模型检测杀菌类农药的腐蚀/刺激性,探讨体外皮肤模型替代农药的动物皮肤安全性评价实验的可行性和影响因素.使用人皮肤成纤维细胞与鼠尾胶原混合培养模拟真皮层,在真皮层上接种人角化细胞,经气液培养形成表皮层.通过细胞活性和细胞因子测定鉴别化合物的腐蚀/刺激性,并比较体外皮肤模型和动物测试结果的一致性.结果表明,皮肤模型对十种农药的腐蚀性测试中,与动物安全评价测试结果完全一致.在刺激性试验中,细胞因子白介素-1a(IL-1a)作为检测终点提高了体外皮肤模型的灵敏性和准确性,人皮肤模型和动物测试结果的一致性达到80％.研究表明,构建的人皮肤模型在对杀菌类农药腐蚀/刺激性测试中初步表现出较好的性能,作为动物替代方法进行农药品安全性评价具有良好的应用前景.     

低质量浓度除草剂2，4滴丁酯处理对土壤动物群落结构的影响

土壤动物是土壤污染的敏感指示生物,对土壤环境变化表现出比较灵敏的反映,为了解低质量浓度除草剂对土壤生态系统的影响,进一步推动除草剂使用的安全性评价研究,采用除草剂2,4滴丁酯对土壤动物群落进行室内染毒模拟试验,参照农田施用常规参考质量浓度,设置4个质量浓度梯度和一个不喷施除草剂的对照质量浓度,质量浓度最高限于参考喷施质量浓度的3~4倍,一次染毒后分别于12、24、48和72 h后对土壤动物进行分离并对群落结构特征进行分析。试验共获得土壤动物18655个,隶属于2门,4纲,12目,群落结构分析结果表明低质量浓度2,4滴丁酯导致土壤动物个体数和类群数有所减少,但减少的程度并不大。个体数量变化与质量浓度增高之间呈现较明显负相关（r=-0.856,P〈0.05）,种类数变化与质量浓度之间不具有明显的相关性,土壤动物种类数的减少主要取决于稀有类群和极稀有类群,数量的变化则主要是优势类群和常见类群的增减。2,4滴丁酯的施加使得土壤动物群落多样性指数和优势度指数降低,均匀度指数增高。土壤动物个体数量和类群数量变化与染毒后经历的时间之间并未表现出明显的规律性,染毒后不同时间土壤动物优势度指数相差较大,而多样性指数和均匀度指数在不同历时时间段内相差不大。综合各分析结果能够看出,低质量浓度2,4滴丁酯处理会对土壤动物群落造成一定影响,但影响程度并不大。     

阿维菌素对中华绒螯蟹肝胰腺氧化胁迫效应和组织结构的影响

采用半静态染毒法,研究不同浓度(1.6、8.0、16.0μg·L-1)阿维菌素在胁迫中华绒螯蟹12、24、48、96 h时肝胰腺超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和丙二醛(MDA)积累量等氧化胁迫相关指标的变化;并在胁迫60 d时,记录各实验组和对照组中华绒螯蟹肝胰腺颜色的变化并进行组织病理观察。3个阿维菌素浓度组为实验组,空白组和溶剂组为对照组。结果表明在整个实验过程中空白组和溶剂组各个时间点各项指标之间无显著差异(P>0.05);低浓度(1.6μg·L-1)组SOD和CAT活力变化不显著(P>0.05);中浓度(8.0μg·L-1)组SOD和CAT活力变化趋势一致,持续被诱导,SOD活力在24 h以后显著高于空白组(P<0.01),CAT活力在48 h以后显著高于空白组(P<0.01);高浓度(16.0μg·L-1)组SOD和CAT活力在12 h~24 h之间变化趋势一致,表现为显著升高(P<0.05),但是在48 h之后,SOD活力显著下降(P<0.05);96 h时,高浓度组SOD活力显著低于空白组(P<0.01),而CAT活力显著高于空白组(P<0.01);在整个实验过程中各实验组MDA积累量逐渐增加,高浓度组氧化胁迫指标的变化幅度大于低浓度组。胁迫60 d时,取样观察可见对照组中华绒螯蟹肝胰腺正常,各实验组中华绒螯蟹肝胰腺发生了不同程度的病变,肝胰腺颜色由最开始的橘黄色变成了淡黄色、黄白色甚至白色,高、中、低各浓度组肝胰腺白化所占的比例分别是66.6%、57.1%和25.0%。组织病理分析显示,淡黄色肝胰腺的病理变化主要是B细胞数量减少,肝胰腺上皮细胞部分肿大并出现空泡;黄白肝病理观察可见肝胰腺上皮细胞中空泡数量增多,体积增大,且空泡内出现内容物,细胞核固缩;白肝的病理表现主要是基膜增厚,上皮细胞大量脱落,散落在管腔中,空泡数量进一步增多,肝胰腺的生理功能受损。由此得出,阿维菌素会对中华绒螯蟹肝胰腺造成氧化胁迫和组织损伤,且阿维菌素和对肝胰腺的氧化胁迫效应和组织结构损伤程度有一定的剂量-效应关系。     

鸡肾脏对微囊藻毒素-LR氧化胁迫的响应

天然蓝藻中蛋白质含量高达40%,与大豆的蛋白质含量相当,是优质饲料蛋白的潜在来源。然而,天然蓝藻中主要含有一种称为微囊藻毒素(Microcystins,MCs)的生物毒素,对动物和人体的健康具有潜在威胁,且关于MCs对家禽氧化胁迫的研究几乎是一片空白。因此,急需开展蓝藻饲料化利用对家禽氧化胁迫的研究,为蓝藻饲料化利用提供理论依据。该文通过腹腔注射的方式研究了崇仁麻鸡肾脏对微囊藻毒素-LR(MC-LR)氧化胁迫的响应。试验设置了5μg·kg~(-1) MC-LR(低剂量组)、10μg·kg~(-1) MC-LR(中剂量组)和20μg·kg~(-1) MC-LR(高剂量组)3个染毒组,同时腹腔注射生理盐水作为对照组。在腹腔注射后的1、3、12、24、48 h检测了鸡肾脏中谷胱甘肽(GSH)含量以及谷胱甘肽转移酶(GST)、谷胱甘肽过氧化物酶(GPX)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性变化以及肾体比变化。结果表明:在MC-LR的作用下,鸡肾脏抗氧化酶(GST、SOD、CAT和GPX)和GSH含量均存在不同程度的变化,但除了3 h时中和高剂量染毒组肾脏中CAT酶活性(值分别为(96.66±1.13)和(74.01±11.20)U·mg~(-1))显著低于对照组(123.58±11.33)U·mg~(-1)(t=4.253;P=0.024和t=4.432;P=0.022)外,其它染毒组的各类抗氧化酶和GSH指标与对照组之间均无显著性差异(P0.05);鸡肾脏系数从1h就开始呈现毒素处理组高于对照组,并在12、24和48 h时,毒素处理组肾脏系数(0.79%-0.87%)显著高于对照组(0.58%-0.70%)的趋势(t=-6.687--3.495;P=0.018-0.036)。上述结果表明,在MC-LR作用下,鸡肾脏产生了氧化应激现象,但是其抗氧化酶系统可能难以抵御MC-LR危害,MC-LR对鸡肾脏的毒性较大。     

氧化乐果对小球藻的毒性研究

以小球藻(Chlorella vulgaris)为实验生物,通过研究不同浓度(0、1、5、10、15、20 mg·L-1)的氧化乐果对小球藻生长发育及EC50,硝酸还原酶活性,叶绿素含量的影响,初步探讨了氧化乐果对小球藻的毒性作用。实验结果表明:氧化乐果对小球藻96 hEC50值为4.88 mg·L-1;低浓度氧化乐果(1 mg·L-1)在前6天对小球藻细胞增长影响不大,高浓度氧化乐果(大于5 mg·L-1)则在初期就表现为抑制,氧化乐果浓度与细胞数量之间表现为负相关(R2=0.65755);在第8天,氧化乐果各浓度组的硝酸还原酶的相对酶活也表现"低促高抑"作用,但是仍然高于对照组的酶活,整体表现为促进作用;随着氧化乐果浓度的增加,小球藻叶绿素含量呈现明显降低的趋势。     

大兴安岭寒温带地区中小型土壤动物群落特征

对大兴安岭北部7个代表性森林群落的中小型土壤动物群落进行调查,揭示该地区中小型土壤动物的区系组成状况、生态分布及动态变化规律.研究区共捕获土壤动物61类, 12 841只,隶属于4门7纲18目44科.其中优势类群4类,即中气门亚目、甲螨亚目、前气门亚目和节跳虫科;常见类群3类,即棘跳虫科、摇蚊科和山跳虫科,两者共占总数量的92.38%.研究结果表明:不同森林群落土壤动物的个体数和类群数分布差异很大,水平分布表现为杨落Ⅱ＞白桦Ⅲ＞桦杜Ⅰ＞蒙古栎Ⅴ＞落沼Ⅳ＞樟子松Ⅵ＞柳草Ⅶ,垂直分布表聚性明显.土壤动物群落的多样性指数(H′)大小为柳草Ⅶ＞白桦Ⅲ＞落沼Ⅳ＞杨落Ⅱ＞桦杜Ⅰ＞樟子松Ⅵ＞蒙古栎Ⅴ.土壤动物个体数的动态变化是10月＞8月＞6月,类群数变化不如个体数变化明显,主要是8月和10月种类较多, 6月较少.多样性指数动态变化表现为8月＞6月＞10月.大多数群落土壤动物的个体数、类群数和多样性指数的动态变化与总体变化趋势相同.图1表5参18     

污染水体中鱼体内三聚氰胺的累积及其在清洁水体中的消除

以吉富罗非鱼（Oreochromis niloticus）为受试动物,在室内用水族箱进行模拟养殖试验,设定污染水体中三聚氰胺初始浓度为0.2 g·L-1,考察罗非鱼在其中饲养1周期间体内三聚氰胺的累积情况。再将鱼转移至不含三聚氰胺的清洁水体中饲养1周,考察鱼体内三聚氰胺在其中的消除情况。结果表明,三聚氰胺能在鱼体内迅速富集,鱼肉及内脏中三聚氰胺的含量均与鱼在污染水体中暴露的时间成正相关。其中,鱼肉中三聚氰胺的浓度几乎成线性增大,拟合所得方程为y=5.598 8x＋12.897,相关系数为0.995 9。内脏对三聚氰胺的富集速率先快后慢,且其中三聚氰胺的浓度为鱼肉的2~3倍。在试验后期,鱼肉及内脏中三聚氰胺的浓度分别高达51.55 mg·kg-1和93.89 mg·kg-1,此时罗非鱼表现出明显的中毒迹象。将罗非鱼转移至清洁水体中养殖后,鱼肉及内脏中三聚氰胺的残留量均迅速下降,尤其是后者的消除速度甚快,一周后两者的浓度分别降至0.13 mg·kg-1和0.21 mg·kg-1。此时,罗非鱼中毒症状逐渐消失,活动趋向正常。     

除草剂苯噻草胺对水稻田土壤微生物种群的影响

以浙江大学华家池校区水稻土为材料,采用室内培养法,研究了不同浓度的外源除草剂苯噻草胺对水稻田土壤可培养微生物种群数量的影响.结果表明:土壤中各微生物类群对不同浓度的苯噻草胺具有各自不同的反应.苯噻草胺能刺激好氧细菌数量增加,但不利于真菌和放线菌的生长.低浓度苯噻草胺刺激水解发酵性细菌(AFB)、产氢产乙酸细菌(HPAB)和产甲烷细菌(MPB)数量的增加,但高浓度苯噻草胺却具有抑制性.在苯噻草胺施用第4周时可强烈刺激反硝化细菌(DNB)数量的增加.苯噻草胺也能刺激厌氧固氮菌(ANFB)数量增加,但这种刺激作用在第2周才出现,而且在培养后期消失.苯噻草胺的短期影响即急性毒性对土壤中可培养微生物的影响较小,比上一代除草剂丁草胺相对安全.图2表1参15     

藻细胞膜电信号对重金属的快速反应研究

为探讨藻细胞膜电信号对重金属离子的响应特征,运用细胞外表面技术和电化学方法研究了淡水藻——大型轮藻(Nitellaflexilis)藻细胞膜电位和膜电阻对汞、镉、铅的快速反应.结果表明,细胞膜电位和膜电阻对汞、镉响应灵敏且快速,30min内即对1μmol·L-1Hg2+、Cd2+表现出超极化和膜电阻增大反应,而5、10μmol·L-1Hg2+、Cd2+则在15min内引起细胞去极化、膜电阻减小,且剂量效应显著.细胞膜电信号对Pb2+的响应浓度为100μmol·L-1,30min内细胞先去极化后超极化,膜电阻持续增大.重金属作用前后相比,高浓度Hg2+、Cd2+(5、10μmol·L-1)导致藻细胞不可逆损伤,而其低浓度所致的损伤可恢复.Pb2+致藻细胞不可逆损伤的最低浓度为500μmol·L-1.对比膜电信号对3种离子的响应特点,发现藻细胞膜电位和膜电阻对Hg2+和Cd2+的响应灵敏度大于Pb2+.     

红松人工林与天然次生林大型土壤动物功能类群

为了解不同森林恢复类型对生态系统物质循环和能量流动的影响,对红松人工林与天然次生林生态系统(硬阔叶林和蒙古栎林)大型土壤动物功能类群时空分布的差异进行分析.在以往研究的基础上将捕获的大型土壤动物划分为腐食性、植食性、捕食性和杂食性土壤动物4个功能类群,并对比红松人工林和天然次生林生态系统土壤动物功能类群.结果表明:(1)水平分布上,天然次生林与红松人工林中4种功能类群大型土壤动物均有分布,其中腐食性土壤动物密度和生物量占绝对优势,但类群组成略有不同;(2)垂直分布上,红松人工林各功能类群土壤动物个体密度及类群数在不同土层差异显著(P0.05),均表现出明显的表聚性;而天然次生林中除腐食性和捕食性土壤动物外,在垂直方向无显著差异(P0.05),0-10 cm层腐食性动物个体密度以及类群数最大;(3)时间变化上,红松人工林与天然次生林土壤动物功能类群表现出一致的变化规律,均以腐食性最高,各类群密度、类群数均为5月、10月高于其他各月,生物量以9月最高.红松人工林和天然次生林土壤动物功能类群时空规律比较研究可为森林经营提供科学依据.     

Metals concentration and bioaccumulation in the marine-coastal trophic web from Buenos Aires province southern coast,Argentina

Metals accumulated in the tissues of organisms inhabiting marine ecosystems generally reflect their bioavailable levels within that environment. The aim of this work was to study the presence and concentration of metals (Cd, Cr, Cu, Fe, Ni, Pb and Zn) and its bioaccumulation in liver-ink sac (n?=?12) and muscle (n?=?12) tissues of loliginid squids, Loligo sanpaulensis, in liver (n?=?15) and muscle (n?=?15) tissues of white croakers, Micropogonias furnieri, and in muscle (n?=?12) tissues of franciscana dolphins, Pontoporia blainvillei, as representatives of different trophic levels of an estuarial ecosystem from the southern coast of Buenos Aires. Except for Pb, all metals were detectable in the hepatic tissues. Bioaccumulation was observed for Fe, Cr and Ni in muscle of squids. Zn levels in muscle samples increased with the trophic level of the species analysed. Necochea-Quequén presents relatively high levels of Ni and Cr in the tissue samples of squids and fishes given other locations of southern hemisphere, indicating a possible anthropic enrichment. These outcomes provide the necessary basis for the environmental monitoring of an area with relevance for fishing and high anthropic activity.     

Role of bioinoculants and organic fertilizers in fodder production and quality of leguminous tree species

The comparative effect of dual inoculation of native N fixer (Rhizobium) and AM fungi consortia with different organic fertilizers (vermicompost and farm yard manure) on fodder production and quality of two leguminous tree species (Leucaena leucocephala (Lam) de. Wit. and Sesbania sesban (L.) Merr.) in silvopastoral system and their impact on the fodder production of un-inoculated Panicum maximum Jacq. under cut and carry system. After three years of plantation maximum tree survival was in L. leucocephala in all the treatments in comparison to S. sesban while fodder production was more in S. sesban for initial two years and in third year it accelerated in L. leucocephala. Dual inoculation with vermicompost significantly improved fodder production, fodder quality and rhizosphere microflora in L. leucocephala but in S. sesban dual inoculation was at par with single inoculation of N fixer, AM fungi and control (without inoculation). The grass production was higher with L. leucocephala for two years while in third year it was more with S. sesban. The association of Rhizobium with AM fungi in L. leucocephala was better than in S. sesban.     

Effect of dietary Jatropha curcas seed meal on rabbits infested by adult ticks of Hyalomma marginatum marginatum

The objective of this study was to investigate the effectiveness of the oral administration of Jatropha curcas seed meal (JCSM) on serum biochemical parameters and histopathology of skin, liver, and kidney of rabbits experimentally infested by Hyalomma marginatum marginatum. Thirty healthy mixed-breed rabbits were randomly divided into five equal groups. The first group was kept as a control and fed soya bean meal. The remaining groups were fed diets containing 2.5%, 5%, 7.5%, or 10% JCSM. At the sixth week, each group was divided into two subgroups. Ticks were introduced to all members of one subgroup, the other subgroup being kept as control. Blood samples were collected and analyzed for biochemical parameters before (zero time) and after the treatment, on the 6th and 8th weeks. Histopathological studies were conducted 8 weeks after the treatment. The results revealed that JCSM could be used in the treatment of ectoparasites at levels of less than 10% in the diet without any serious effects on liver and kidney functions.     

Effects of alcoholic extract of Momordica charantia (Linn.) whole fruit powder on the pancreatic islets of alloxan diabetic albino rats

Alcoholic extract of whole fruit of Momordica charantia was prepared. Adult healthy albino rats were divided into four groups and received a dose of 6 mg/l00 gm. body weight of alloxan monohydrate. Animals of group I served as diabetic control group. The animals of II, III, and IV groups received 25 mg, 50 mg and 75 mg doses of the extract respectively for different durations. 75 mg dose showed increase in body weight. All doses of alcoholic extract of M. charantia were able to decrease the blood sugar level significantly. Extract feeding showed definite improvement in the islets of Langerhans. No toxic effect was observed in the liver The significant features of the study have been blood glucose once lowered by the treatment with M. charantia fruit extract remained static even after discontinuation of drug for 15 days. Blood sugar never fell below normal values even with a high dose, in pancreatic islets, beta cells showed definite improvement.     

Toxicological studies of zinc oxide nanomaterials in rats

In this study, we have evaluated the ability of zinc oxide (ZnO) nanoparticles to induce pulmonary and extrapulmonary toxicities was examined in rats following intratracheal (IT) instillation. Lungs of rats were instilled IT with either phosphate-buffered saline (PBS)?+?1% Tween 80, ZnO nanoparticles, carbonyl iron or quartz particles at a dose of 1 or 5?mg?kg^?1 body weight. Following exposure, bronchoalveolar lavage (BAL) fluid, blood samples and organs including lung, liver, kidneys, heart, pancreas, and brain were collected at 24?h, 1 week, or 1 month of post instillation of nanoparticles and different parameters estimated to assess toxicity. BAL fluid was analyzed for lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) to assess pulmonary toxicity. Exposures to ZnO or quartz particles produced transient dose-dependant increase in BAL fluid LDH and ALP activities at all post exposure periods. Blood samples were analyzed for the tissue damage biomarkers to assess extrapulmonary toxicity. Histopathological examination of lung, liver and kidneys revealed dose-dependent degeneration and necrosis which worsened at 1 week post-instillation periods but recovered at 1 month post instillation. Histopathological examination of rat pancreas, heart, and brain exposed to quartz or ZnO particles showed no marked changes. Data suggest the instillation of ZnO nanoparticles produced a greater pulmonary toxicity in rats comparable with quartz; and extrapulmonary toxicities of these ZnO nanoparticles might be due to translocation into liver and kidney.     

Potential correlation between perfluorinated acids and liver morphology in East Greenland polar bears (Ursus maritimus)

In this study, residual concentrations of chlorpyrifos (CPF) were determined in feed (40) and fodder (25) samples collected from various locations of Tarai region of Uttarakhand. For extracting residues, liquid–liquid partition followed by alumina column clean up was used and the detection and quantification of residues was undertaken with the help of high-performance liquid chromatography using C18 column and diode array detector at 220?nm. Of the total 40 feed samples analyzed, 7 (17.5%) samples were found positive for CPF with the mean residual concentration of 0.058?µg?g^?1; while out of 25 fodder samples, CPF residues were detected in a single (4%) sample with residual concentration of 0.39?µg?g^?1. However, none of the feed or fodder samples contained CPF residues above the prescribed limit.     

Influence of temperature on muscle-fibre development in larvae of the herringClupea harengus

The development of swimming (myotomal) muscles was studied in herring larvae (Clupea harengus L.) caught in the Firth of Clyde, Scotland, in spring 1990 and reared at either 5, 10 or 15°C. Two muscle-fibre types can be distinguished in the myotomes of herring larvae using ultrastructural criteria. A single layer of small-diameter muscle fibres, packed with mitochondria, is present beneath the entire surface of the skin (superficial muscle fibres). The remaining bulk of the muscle is composed of larger diameter fibres (inner muscle fibres) containing significantly more myofibrils than the superficial fibres. In 1 d-old larvae, the number of inner muscle fibres in myotomes immediately posterior to the yolk-sac was 311±41 at 15°C, 257±22 at 10°C and 187±22 at 5°C (mean±SD,n=6). The average diameter of inner muscle fibres increased with decreasing temperature, so that the total cross-sectional area of muscle was similar at each temperature. After 6 to 7 d, the number of muscle fibres had significantly increased at 15°C (383±25), but not at 10°C (281±32) or 5°C (192±17). In contrast, the average cross-sectional area of inner muscle fibres had increased by 19% at 15°C, 34% at 10°C, and 26% at 5°C. Temperature also influenced the relative proportions and spatial distributions of muscle-fibre organelles. For example, in 1 d-old larvae, the fraction of muscle-fibre volume (volume density) occupied by mitochondria in the superficial fibres was significantly higher at 15°C (46.0%) than at either 5°C (37.6%) or 10°C (38.8%). In the inner muscle fibres, the volume density of mitochondria was 26.1% at 15°C, 20.5% at 10°C and 15.9% at 5°C, whereas the volume density of myofibrils was similar at the three temperatures (33 to 38%). Typically, inner muscle fibres from 10°C larvae, but not from 5 or 15°C larvae contained a large central mitochondrion.     

Evaluation of fodder quality of straw of different landraces of rice (Oryza sativa L.) under cultivation in the Kumaun Region of Indian Central Himalaya

Thirty landraces of rice collected from various locations, across an elevational gradient (1150–2400 m asl), in Kumaun, Central Himalaya and VL-206 (a high-yielding variety recommended for rainfed mountain area) have been evaluated, both in terms of quantity and nutritional quality of straw. Data were collected to estimate the percentage share of fodder in the overall biomass produced. The total biomass harvested at maturity was taken as productivity. In view of the severe scarcity of fodder in hill regions, particularly during winter months, straw was also examined for quality-linked biochemical parameters and dry matter digestibility (DMD) of fodder. The Crude Protein (CP) values ranged from 2.21% (Nandhani) to 6.33% (Dalbadal), and in saceo DMD (nylonbag technique) was highest in Kaladur (48.84%) and lowest in Kantolia (32.25%). While the CP in VL206 was within the range recorded for various landraces (3.61%), DMD value (27.29%) was lower than that of all the landraces tested. In saceo DMD data indicate that the fodder quality of landraces is better than that of the introduced variety. The values for neutral detergent fibre (NDF) ranged from 63.64% (Sabhawati) to 78.46% (Chhotiya) among the rainfed landraces, while in the irrigated landraces, Thapchini had more NDF (87.50 %), as compared to Kantolia (73.88%) and VL206 (74.91%). Values for the acid detergent fibre (ADF) varied from 42.70% (Danbasmati) to 54.58% (Nandhani) in the rainfed group; ADF was slightly higher in Thapchini (49.41%) in comparison to Kantolia (47.19) and VL206 had the highest (56.80%) ADF content. The observed differences with respect to straw productivity, chemical composition and in saceo DMD in various landraces, even when grown under similar conditions at one place, clearly indicate the richness of the gene pool of rice being maintained, through cultivation, in the Kumaun hills. It may be mentioned that agriculture in the hills is biomass based, and livestock form an integral component of subsistence farming.     

Influence of development and rearing temperature on the distribution,ultrastructure and myosin sub-unit composition of myotomal muscle-fibre types in the plaice Pleuronectes platessa

Eggs of the plaice Pleuronectes platessa L. were incubated at temperatures of 5, 8, 10, 12 and 15°C in March 1990, 1991 and 1992. The myotomes of yolk-sac larvae contain a single superficial layer of small-diameter muscle fibres which stain intensely for succinic dehydrogenase activity, surrounding 390 to 500 weakly staining inner-muscle fibres of larger diameter. Larvae reared at 15°C only survived for a few days and had significantly more inner-muscle fibres of larger average cross-sectional area than those hatching at 5 to 10°C. Myofibrils occupied 61% of the volume of inner-muscle fibres in 15°C larvae compared with 35 and 36% in larvae hatching at 5 and 10°C, respectively (P(0.01). Following metamorphosis, which occurs between 7 and 10 wk, the myotomes retain the single layer of superficial-muscle fibres characteristic of larvae. A thickening of the superficial-muscle layer is first evident in 4 to 5 mo-old laboratory-reared fish of 20 mm total length (TL) and in 0-group fish caught in June and July. On the basis of the histochemical staining reactions for myofibrillar ATPase and succinic dehydrogenase activities, the myotomes of 1-group (104 mm TL) and adult (280 mm TL) plaice were found to contain a minimum of six distinct muscle-fibre types. Two-dimensional gel electrophoresis and peptide mapping were used to investigate changes in myosin subunit composition during development. Myosin from the inner muscle of larvae contains two isoforms of the phosphorylatable light-chain 2 (LC2_L1 and LC2_L2). Following metamorphosis and during the first year, inner-muscle fibres co-express LC2 isoforms characteristic of the superficial fast-muscle fibres of adult plaice (LC2_F1 and LC2_F2) in addition to the larval isoforms. Fast-muscle fibres isolated from deep layers of the myotomes in adult plaice only contain LC2_F2. In contrast, myosin from larval muscle and adult fast muscle contain apparently identical alkali light chains (LC1 and LC3). Peptide maps of myosin heavy chains (MHCs) from 6 wk-old larvae and 10 wk-old fish that had completed metamorphosis are similar, but distinct from those of 1-group plaice. Further changes in white-muscle MHC composition are evident between 1-group fish of 104 mm TL and adults of 280 mm TL.     

Temperature and development in larvae of the turbot Scophthalmus maximus

Turbot (Scophthalmus maximus L.) were reared at 12 and 16°C until 26 d after hatching. At both temperatures, starting at the neural plate stage, somites were initially formed every 75 min. Expressed as a percentage of development time (DT, fertilisation to 90% larvae hatching) somite formation occurred relatively earlier during embryogenesis at 12°C (45% DT) than at 16°C (55% DT). At 12°C, after the 32-somite stage the rate of somite formation decreased to one every 300 min. The larvae hatched after 6 d at 12°C and 3 d at 16°C at a relatively primitive stage of development, prior to the opening of the mouth and anus, with unpigmented eyes, and a straight gut. Temperature altered the relative timing of organogenesis in the larval stages. At 12°C, the following characters appeared (in this order): swimbladder>loop in the gut (at the time of yolk exhaustion)>caudal fin. In contrast, at 16°C, the caudal fin appeared at the same time as the loop in the gut. At 16°C, spines formed on the head in the region of the otic capsule at the time the swimbladder formed and the yolk was exhausted, but were absent in 12°C larvae. At both temperatures, in 1 d-old larvae the myotomes just behind the yolk-sac contained 200 inner muscle fibres (presumptive white muscle). The initial growth of inner muscle was largely due to hypertrophy, but by 26 d at 12°C and 11 d at 16°C hyperplastic growth became important, as evidenced by a significant increase in the number of small fibres (<10 m²). By 26 d the average number of inner muscle fibres had increased to 341 at 12°C and 988 at 16°C. New muscle fibres were added in distinct germinal zones at the dorsal and ventral apices of the myotomes. Metamorphosis was associated with a thickening of the superficial (presumptive red) muscle layer and the appearance of tonic muscle fibres.