基于机器视觉的鱼类模式生物在线监测技术方法研究

水污染的防治问题是我国关注的重中之重,现有理化监测方法的实时性和综合性较差,特别是对于一些极端可变化的环境,更需要新的方法以辅助和解决。生物式水质监测方法被提出,通过利用生物对环境污染或变化所产生的反应来直接或间接体现水质的污染情况。然而,观测指标与量化标准是面临的一大难题。文章利用机器视觉的方法,以青鳉鱼为模式生物,并以青鳉鱼的生理特征以及运动特征(呼吸频率、胸鳍摆动频率、摆尾频率)为观测指标,两方面综合评定青鳉鱼应激状态,实时监测与分析。实验结果表明该方法能为生物式水质监测和预警的发展提供一定支持与参考。测得青鳉鱼呼吸频率为3.06 Hz,胸鳍摆动频率为4.83 Hz,尾鳍摆动频率为5.08 Hz,结果与实际指标一致。     

一种计量青鳉鱼胸鳍和尾鳍摆动频率和幅值的计算机视觉算法

青鳉鱼在饮用水安全领域具有很高的应用价值,其鱼鳍的活动状态可以反映水质的污染状况。但由于技术条件的限制,传统的方法无法实时而有效地提取到这些特征。针对这个问题,提出了一种可以高效识别并测量青鳉鱼胸、尾鳍摆动频率和幅值的计算机视觉算法。其中,为了快速地提取到这2个细节,自动阈值分割、帧差法、背景差分法、重心法、图像卷积和骨架细化等耗时极短的图像处理方法被应用到这项研究当中。初步试验显示这种算法在应用过程中是高效而可行的。它可以被广泛地应用到水环境监测领域,如研究有毒物质对青鳉鱼行为特性的影响和评估水环境的危险程度。     

水环境中重金属铜暴露下青鳉鱼的行为响应

采用生物行为传感器监测青鳉鱼在重金属铜暴露下的行为数据,分析不同暴露浓度(20、10、5、1和0.1 TU)下青鳉鱼的行为响应。20、10、5、1和0.1 TU的暴露浓度下青鳉鱼对重金属污染的行为反应模式符合环境胁迫阈值模型,且不同浓度梯度重金属对青鳉鱼产生不同的行为毒性效应。利用重金属作用下青鳉鱼的行为变化来研究重金属环境胁迫导致的生物行为响应机制,从而得到重金属暴露下生物行为的实时变化过程和趋势,可对水环境生态系统质量进行综合评价。     

水环境中重金属铜暴露下青鳉鱼的行为响应

本研究采用生物行为传感器监测青鳉鱼重金属铜暴露下的行为数据，分析不同暴露浓度(20、10、5、1和0.1 TU)下青鳉鱼的行为响应。20、10、5、1和0.1 TU的暴露浓度下青鳉鱼对重金属污染的行为反应模式符合环境胁迫阈值模型，且不同浓度梯度重金属对青鳉鱼产生不同的行为毒性效应。通过本研究，利用重金属作用下青鳉鱼的行为变化来研究重金属环境胁迫导致的生物行为响应机制，从而得到重金属暴露下生物行为的实时变化过程和趋势，可对水环境生态系统质量进行综合评价。     

溶解性有机碳的主要组成对青鳉鱼铜急性毒性的影响

为揭示溶解性有机碳(DOC)的组成对生物配体模型(BLM)预测铜的生物毒性准确性的影响,研究了不同浓度黄腐酸(FA)、腐殖酸(HA)和两者不同浓度比例混合物影响铜对青鳉急性毒性。结果表明:在相同水质条件下,黄腐酸或腐殖酸浓度增加时,铜对青鳉的LC50均增加;两者共存时,当腐殖酸质量百分比从10%增至90%时,铜对青鳉的LC50也增加。当天然水中DOC的组成不确定时,可假设HA和FA的组成比例为1∶1,此时BLM预测铜对青鳉的LC50的偏差最小。     

铜在日本青鳉体内的累积特征及其对肝脏组织结构的影响

为了解水体铜(Cu(Ⅱ))在日本青鳉(Oryzias latipes)脑、鳃、肝脏、性腺和肌肉组织中的累积特征,以及Cu(Ⅱ)对肝脏组织结构的影响,本文采用生态学单因子梯度实验,进行20 d的日本青鳉毒性暴露试验。结果显示,日本青鳉各个组织对Cu(Ⅱ)的累积量随着浓度的升高而增多,但不同组织对Cu(Ⅱ)的累积量有较大的差异,铜累积量由大到小依次为:肝脏脑鳃性腺肌肉。铜暴露后的日本青鳉肝细胞出现肿大、大小不一、排列混乱,静脉扩张淤血,部分肝细胞固缩和坏疽等现象。肝脏是日本青鳉蓄积铜的主要器官之一,实验发现高浓度的Cu(Ⅱ)对肝脏组织结构造成了明显的损伤,进而严重影响日本青鳉的存活率。     

2种除草剂联合胁迫下日本青鳉的逐级行为响应(The Stepwise Behavioral Responses of the Japanese Medaka under the Joint Stress of Two Herbicides)

探讨了日本青鳉在两种除草剂阿特拉津和百草枯联合暴露下的逐级行为响应,采用水质安全在线生物预警系统(BEWs)记录行为强度数据,分析不同暴露浓度、不同暴露时间日本青鳉的行为响应.10、5、1和0.1TU的暴露浓度下行为反应时间分别为:0.74、7.7、29.4和42.2h,而每个浓度不同配比之间行为反应时间差异明显.结果表明:在两种除草剂的暴露下,日本青鳉的逐级行为响应既受化合物浓度高低的影响,又受暴露时间的影响,而且每个浓度两种药物不同配比暴露下的青鳉鱼的逐级行为响应基本一致,每个浓度不同配比之间行为反应时间差异明显,即两种作用机制不同的除草剂对日本青鳉的行为毒性是协同作用.     

青鳉鱼的行为特征提取研究

采用生物行为传感器获取青鳉鱼的行为数据时,青鳉鱼个体差异会导致采集到的原始电信号在时空特性下完全不同。重要的行为特征往往被隐藏在原始信号中,传统的信号处理方法无法实时而有效地提取到这些特征。针对这个问题,观察并记录了暴露实验前后青鳉鱼的行为变化,提出了一种可以高效表征行为特征的直方图统计算法。实验结果表明,该方法能够准确对应人眼观测到的暴露实验前后鱼的行为变化趋势,同时也为后续异常行为识别提供一定的支持和参考。     

应用生物配体模型研究湘江水体中铜的生物有效性

为评估湘江水体中铜的生物有效性,通过铜对青鳉的急性毒性实验确定铜对青鳉的半致死浓度(LC₅₀),然后利用生物配体模型(Biotic Ligand Model, BLM)确定青鳉的累积半致死浓度(LA₅₀)。同时对湘江水样的预测结果表明,铜对青鳉的LA₅₀为12.43 nmol?gw^-1;BLM预测的与实验观测的LC₅₀差异倍数小于2,表明BLM预测结果非常准确。在此基础上预测铜对青鳉LC₅₀预测范围为344.70～761.96 μg?L^-1。湘江各点水质最终急性值(FAV)相差不大(11.04～14.25 μg?L^-1),但水中溶解态铜含量相差较大(0.29～10.48 μg?L^-1),同时毒性单位值(TU)相差较大(0.06～1.48 μg?L^-1),其中最小值在舂陵水入湘江下游采样断面,最大值在株洲断面。在全部调查断面中,只有株洲断面TU大于1(1.48),说明株洲断面水中铜含量超过了该断面水中BLM确定的标准最大浓度1.48倍。     

镉胁迫下黑点青鳉(Oryzias melastigma)肝脏金属硫蛋白mRNA性别差异性表达研究

鱼类金属硫蛋白(metallothionein,MT)mRNA作为生物标志物已越来越多地应用到水环境重金属污染的监测中,然而重金属胁迫下鱼类MT mRNA表达是否具有性别差异性特征目前尚无相关报道。以海洋模式鱼类—黑点青鳉(Oryzias melastigma)为实验鱼类,首次克隆了其MT基因全长c DNA序列,分析了黑点青鳉MT基因和氨基酸序列特征,然后进一步研究了水体中镉暴露后雌、雄性黑点青鳉肝脏中MT mRNA表达的差异性。结果显示,黑点青鳉MT基因c DNA全长为385 bp,编码60个氨基酸,预测编码的多肽分子量为5 990 Da,含有脊椎动物MT基因特征性序列结构cys-x-cys、cys-x-x-cys和cys-x-cys-cys。镉胁迫下雌、雄性黑点青鳉肝脏MT mRNA表达结果显示,在0.8,4.0和8.0μg·L-1环境浓度的镉暴露1~7 d条件下,与对照组相比,雄性黑点青鳉在所有的不同暴露浓度和不同暴露时间的实验组中肝脏MT mRNA几乎均被显著性诱导表达;而雌性黑点青鳉只在较高暴露浓度和较长暴露时间的实验组中才被显著性诱导表达,并且MT mRNA的表达水平比雄性低得多。上述研究结果表明,在相同的镉胁迫条件下,雄性黑点青鳉肝脏MT mRNA的表达比雌性更敏感;以黑点青鳉MT mRNA作为生物标志物监测海洋环境重金属污染生物效应状况时,应选择更敏感的雄性黑点青鳉作为监测鱼类。利用生物标志物监测水环境污染物生物效应时,监测动物的性别差异是一个应当考虑的重要因素。     

Consequences of paternal care on pectoral fin allometry in a desert-dwelling fish

Positive static allometry is a scaling relationship where the relative size of traits covaries with adult body size. Traditionally, positive allometry is thought to result from either altered physiological requirements at larger body size or from strongly condition-dependent allocation under sexual selection. Yet, there are no theoretical reasons why positive allometry cannot evolve in fitness-related traits that are solely under the influence of natural selection. We investigated scaling and sexual dimorphism of a naturally selected trait, pectoral fin size, in comparison to a trait important in male–male combat, head width in natural populations of a fish, the desert goby Chlamydogobius eremius. Male desert gobies provide uniparental care and use their pectoral fins to fan the brood (often under hypoxic conditions); hence, larger fins are expected to be more efficient. Male pectoral fins do not appear to fulfil a signalling function in this species. We found that, for both pectoral fin size and head width, males exhibited positive allometric slopes and greater relative trait size (allometric elevation) than females. However, for head width, females also showed positive allometry, albeit to a lesser degree than males. Because fin locomotory function typically does not result in positive allometry, our findings indicate that other naturally selected uses, such as paternal care, can exaggerate trait scaling relationships.     

Osteological development of the vertebral column and of the fins in Diplodus sargus (Teleostei: Perciformes: Sparidae)

The osteological development of vertebral column and fins in white sea bream, Diplodus sargus (Linnaeus, 1758) was studied. Vertebral ontogeny started at 5.7 mm total length (TL, post-fixation), with the formation of the first cartilaginous neural and haemal arches, and was completed by the full attainment of dorsal ribs (20.8 mm TL). The formation of vertebral centra occurred between 7.3 and 9.4 mm TL, but was fully completed only after the development of post- and pre-zygapophyses (26.0 mm TL). Pectoral supports were the first fin elements that started to develop (3.8 mm TL), followed by those of the caudal fin (5.2 mm TL), dorsal and anal fins (6.7 mm TL), and then by those of the pelvic fin (9.5 mm TL). The caudal fin of D. sargus was the first to develop fin rays and attain the full count of lepidotrichia (5.6-7.8 mm TL), but the last to attain the full count of dermatotrichia (7.9-16.0 mm TL). The next fins starting to present rays were the pectoral (6.5 mm TL) and the dorsal and anal fins (7.5 mm TL), while the pelvic fins were the last (11.5 mm TL). Following the caudal lepidotrichia (7.8 mm TL), the anal (10.5 mm TL), dorsal (12.0 mm TL), pectoral (11.4 mm TL) and pelvic fins (12.5 mm TL) were the next with fully completed ray counts. Finally, fin meristics were fully developed with the caudal dermatotrichia. The results are discussed in comparison with the fin and vertebral ontogeny of other teleosts.     

氯化镉、灭多威和硝基苯对日本青鳉(Oryzias latipes) 行为抑制的生物应急监测

研究了氯化镉、灭多威和硝基苯对孵化以后7d左右日本青鳉(Oryzias latipes)行为毒性。结果表明,在不同污染物中,随暴露浓度的增加,第1尾日本青鳉产生行为抑制的时间逐渐缩短,并且半数行为抑制时间(time ofhalf behavior inhibition,THBI,Y)随暴露浓度(X)呈现幂指数关系Y=aXb,其中20相似文献   

Osteological development of Dentex dentex (Osteichthyes: Sparidae): dorsal, anal, paired fins and squamation

The osteological development of dorsal, anal and paired fins and the squamation in common dentex, Dentexdentex (Linnaeus, 1758), was studied under extensive larval rearing conditions. The ontogeny of the fins was determined in three stages: (1) the yolk-sac and the early larval stage (up to 6.7 mm in vivo total length, TL), in which the majority of the elements of the pectoral fins and the predorsals, dorsal and anal proximal pterygiophores formed; (2) the late larval stage (up to 11.6 mm TL), in which the majority of the pectoral lepidotrichia, the lower extrascapulae, all the elements of the pelvic fins, as well as the lepidotrichia, spines and the distal radials of the rest fins developed; and (3) the metamorphosis stage (up to 24.0 mm TL), when the upper extrascapulae formed and the pectoral lepidotrichia attained their full count. The ontogeny of squamation was carried out during the metamorphosis stage; it started at 13.4 mm TL with laterally symmetric points on the middle of each side of the body, followed by one ventral centre (15.6 mm TL) which was located anterior to the pelvic fins, and two bilateral pairs of centres which appeared on the head (17.8 and 21.2 mm TL). The results are discussed from a functional viewpoint and in comparison with the fin and scale ontogeny of other teleosts. Received: 1 March 2000 / Accepted: 20 September 2000     

Migratory history of the Russian sturgeon Acipenser guldenstadti in the Caspian Sea, as revealed by pectoral fin spine Sr:Ca ratios

Ontogenic patterns of change in pectoral fin spine Sr:Ca ratios were examined in the Russian sturgeon Acipenser guldenstadti from the Caspian Sea. Pectoral fin spine Sr:Ca ratios of the sturgeon fluctuated strongly along the life history transect in accordance with the migration pattern from freshwater to sea (brackish) water habitats, i.e. all specimens exhibited a typical anadromous pattern in the ratio. Several specimens showed two transition points in pectoral fin spine Sr:Ca ratios from low ratios to high, indicating that those specimens had a flexible migration strategy in the ambient water, and could migrate downstream to the Caspian Sea multiple times after spawning.     

我国海水水质基准的构建:以三丁基锡为例

我国近岸海域污染形势严峻、污染物种类繁多、生态健康风险复杂,现有的海水水质标准难以满足当前海洋环境污染评价、管理和监控的需求,亟需适宜我国海洋区系特征的海水水质基准作为水质标准修订与制定的理论依据.借鉴美国、欧盟和荷兰等国家水质基准的制定方法,提出了构建我国海水水质基准的思路,并给出了数值型双值基准的数据要求、定值方法和相应的计算模式等.以三丁基锡化合物(TBT)为例,按照论文建立的方法,计算了保护我国海洋水生生物的TBT基准.结果表明:我国TBT化合物海水水质基准高值(HSWC)和海水水质基准低值(LSWC)分别为0.43μg·L-1和0.002μg·L-1,与美国,加拿大和英国等国家的推荐基准值略有不同.研究结果将为我国海水水质基准的制定与研究提供帮助.