Skin biopsy of Mediterranean cetaceans for the investigation of interspecies susceptibility to xenobiotic contaminants.

Various studies on Mediterranean cetaceans have revealed bioaccumulation of contaminants such as organochlorines (OCs) and heavy metals. The susceptibility of these animals to organic pollutants and the relationship between bioaccumulation and population decline (as in the case of Delphinus delphis) are unexplored fields. In this study, we used a non-destructive approach (skin biopsy) to explore OC bioaccumulation processes and mixed-function oxidase activity (BPMO) in four species of cetaceans: striped dolphin (Stenella coeruleoalba), bottlenose dolphin (Tursiops truncatus), common dolphin (D. delphis) and fin whale (Balaenoptera physalus). Significant differences in BPMO induction and OC levels were found between odontocetes and mysticetes, the former having mixed-function oxidase activities four times higher than the latter, binding with levels of OCs one order of magnitude higher in odontocetes. A significant correlation (P < 0.05) between BPMO activities and OC levels was found in B. physalus. In an ongoing project, fibroblast cultures have been used as an alternative in vitro method of evaluating interspecies susceptibility to contaminants such as OCs and polycyclic aromatic hydrocarbons (PAHs). These results suggest that cetacean skin biopsies are a powerful non-invasive tool for assessing ecotoxicological risk to Mediterranean marine mammals species.     

Use of immunofluorescence technique in cultured fibroblasts from Mediterranean cetaceans as new "in vitro" tool to investigate effects of environmental contaminants

The aim of the present study was to propose the immunofluorescence technique in cultured fibroblasts from Mediterranean cetaceans as a new "in vitro" tool to explore the susceptibility of these marine mammals to different xenobiotic compounds. The cell lines were cultured from integument biopsies of free-ranging and stranded cetaceans (dead within 12h). Using the indirect immunofluorescence assay, we detected endogenous proteins induced by different contaminants. Here we present the method used for qualitative and quantitative evaluation of cytochromes P450 (CYP1A1 and CYP2B) induced by some POPs (DDTs and PCBs) and emerging contaminants (PBDEs) in fibroblast cell cultures of striped dolphin (Stenella coeruleoalba) and bottlenose dolphin (Tursiops truncatus). Immunofluorescence was quantified with a specially designed Olympus macro, DetectIntZ. A major result was the possibility of using this "in vitro" assay to quantify induction of endogenous proteins.     

First detection of CYP1A1 and CYP2B induction in Mediterranean cetacean skin biopsies and cultured fibroblasts by Western blot analysis

The need to develop powerful tools to detect exposure and effects of POPs and emerging contaminants in Mediterranean cetaceans led us to develop a suite of sensitive non-lethal biomarkers in integument biopsies of free-ranging animals. In order to propose induction of CYP1A1 and CYP2B, detected by Western blot analysis, as biomarkers of exposure to OCs, PAHs and PBDEs, a three-phase experimental protocol (in vitro experiments, calibration experiments and field applications) was followed using fibroblast cell cultures and biopsies of Mediterranean Stenella coeruleoalba and Tursiops truncatus. This methodology was confirmed to be sensitive and stable in comparison to previous methods used to detect CYP1A1 in biopsies, enabling analysis of several inducible proteins in non-lethal samples and analysis of material from stranded animals.     

Assessment of toxicological status of a SW Mediterranean segment population of striped dolphin (Stenella coeruleoalba) using skin biopsy

Various studies have revealed high concentrations of contaminants such as organochlorines (OCs) and heavy metals in Mediterranean cetaceans. A geographical trend of contamination (PCBs and DDTs) has been found for striped dolphin (Stenella coeruleoalba). In this study we used a non-lethal approach (skin biopsy) to investigate bioaccumulation of OCs, including polychlorobiphenyls (PCBs), DDTs, polychlorodibenzo-p-dioxins (PCDDs), polychlorodibenzofurans (PCDFs), trace elements (Hg, Cd, Pb) and CYP1A activity (BPMO) in nine striped dolphins sampled in the Aeolian area (Sicily - Italy) in summer 2002. The arithmetic mean value of BPMO activity in this group was 43.46 AUF/g tissue/h. This value is approximately 3 times and 5 times lower, respectively, than the value found in the Ionian and in the Ligurian groups. Skin biopsies of striped dolphins emerged as a suitable material for assessing the toxicological status of the various Mediterranean groups.     

Assessment of toxicological status of a SW Mediterranean segment population of striped dolphin (Stenella coeruleoalba) using skin biopsy

Various studies have revealed high concentrations of contaminants such as organochlorines (OCs) and heavy metals in Mediterranean cetaceans. A geographical trend of contamination (PCBs and DDTs) has been found for striped dolphin (Stenella coeruleoalba). In this study we used a non-lethal approach (skin biopsy) to investigate bioaccumulation of OCs, including polychlorobiphenyls (PCBs), DDTs, polychlorodibenzo-p-dioxins (PCDDs), polychlorodibenzofurans (PCDFs), trace elements (Hg, Cd, Pb) and CYP1A activity (BPMO) in nine striped dolphins sampled in the Aeolian area (Sicily – Italy) in summer 2002. The arithmetic mean value of BPMO activity in this group was 43.46 AUF/g tissue/h. This value is approximately 3 times and 5 times lower, respectively, than the value found in the Ionian and in the Ligurian groups. Skin biopsies of striped dolphins emerged as a suitable material for assessing the toxicological status of the various Mediterranean groups.     

Assessment of cytochrome P450 1A in harbour seals (Phoca vitulina) using a minimally-invasive biopsy approach

Biomarkers of organochlorine exposure, such as the induction of cytochrome P450 1A (CYP1A), can be used to assess the impact of environmental contaminants on the health of free-ranging marine mammal populations. The objective of the present study was to measure CYP1A in skin and liver biopsies obtained from live harbour seals (Phoca vitulina). Twelve harbour seal pups, aged three to five weeks, were captured from the Fraser River estuary, British Columbia, Canada, and temporarily held in captivity. Skin ( approximately 60 mg) and liver ( approximately 40 mg) biopsies, obtained while seals were under general anaesthesia, yielded sufficient tissue for the measurement of CYP1A by immunoblot analysis and ethoxyresorufin O-deethylase activity. A short-term exposure experiment, in which harbour seals (n=3) were treated orally with beta-naphthoflavone (BNF), resulted in increased hepatic and cutaneous CYP1A protein levels, consistent with observations in other mammals. This study is the first to measure CYP1A in skin and liver biopsies from live harbour seals and to report in vivo BNF-associated CYP1A induction in a marine mammal. The results demonstrate that microsamples collected using minimally-invasive techniques can provide toxicologically-relevant information form marine mammals.     

Ten guiding principles for the delineation of priority habitat for endangered small cetaceans

The adoption of endangered species laws in various nations has intensified efforts to better understand, and protect, at-risk species or populations, and their habitats. In many countries, delineating a portion of a species' habitat as particularly worthy of protection has become a mantra of these laws. Unfortunately, the laws themselves often provide scientists and managers with few, if any, guidelines for how to define such habitat. Conservationists and scientists may view protecting part of the habitat of an endangered species as an ineffectual compromise, while managers may be under pressure to allow a range of human activities within the species' habitat. In the case of small cetaceans, establishing boundaries for such areas can also be complicated by their mobility, the fluid nature of their environment, and the often ephemeral nature of their habitat features. The convergence of multiple human impacts in coastal waters around the world is impacting many small cetaceans (and other species) that rely on these areas for feeding, reproducing, and resting. The ten guiding principles presented here provide a means to characterize the habitat needs of small, at-risk cetaceans, and serve as a basis for the delineation of ‘priority habitat’ boundaries. This conceptual approach should facilitate a constructive discourse between scientists and managers engaged in efforts to recover endangered species. The degree to which the recovery of an at-risk species can be reconciled with sustainable economic activity will depend in part on how well these principles are incorporated into the delineation of priority habitat.     

Spatial and temporal assessment of potential risk to cetaceans from static fishing gears

Ecosystem Based Fisheries Management (EBFM) requires the consideration of potential impacts of a commercial fishery on all components of the ecosystem. Assessment of the impact of commercial fishing on marine mammals generally focuses on species at known risk from bycatch. For cetaceans in particular, inclusion under the Threatened, Endangered and Protected (TEP) species component of Ecological Risk Assessment for the Effects of Fishing (ERAEF) can seem redundant if a species is already known to be at risk or is not thought to interact with the fishery with consequences for its conservation. A spatially and temporally explicit Productivity Susceptibility Analysis (PSA) procedure was developed for inclusion under ERAEF to allow cetacean species to be screened for risk. The technique is demonstrated by assessing the potential risk to harbour porpoise and minke whales from a number of static gear fisheries. The results demonstrate that although a fishery might pose high risk to a species, low or moderate risk areas can exist within the range of the fishery, enabling management measures to focus on areas of greatest risk. Designed to complement and support existing methods of bycatch assessment, this approach is a repeatable and standardised assessment, the outputs of which can be used to systematically document the level of risk posed to different species in a transparent way to aid the inclusion of cetaceans in ERAEF and EBFM both now and in the future.     

Using trace elements in skin to discriminate the populations of minke whales in southern hemisphere

Concentrations of 12 trace elements (V, Cr, Mn, Cu, Zn, Se, Rb, Sr, Cd, Cs, Ba, and Hg) were determined in liver and skin tissues of minke whales from various regions within the Antarctic Ocean. Cd concentrations in livers of southern minke whale were apparently higher than those in cetaceans from other regions, while Hg concentrations were lower. There were significant positive correlations between body length and concentrations of Cd and Hg in the liver. The concentrations of all trace elements in the skin were lower than those in other cetaceans reported previously. Significant positive correlations between liver and skin were found for Cr, Mn, Cu, Zn, Rb, Cd, and Cs, implying that the concentrations of these trace elements in the skin reflect those of internal organs. Large interannual variation of the accumulation pattern of trace elements in the skin was observed for the southern minke whales from Area V. There were significant differences in the skin element concentrations among Areas III, IV, and V, especially for males. Also, discriminant analysis between geographically two different groups collected during 1995/1996 austral summer season, based on the concentrations of trace elements in the skin, allowed for a correct classification of 90% of these minke whales. These results suggest that measurement of trace elements in skin samples could provide valuable information on the status of contamination and possible geographic differences in the accumulation levels in southern minke whales.     

Forecasting the consequences of climate-driven shifts in human behavior on cetaceans

While climate change is expected to affect cetaceans primarily via loss of habitat and changes in prey availability, additional consequences may result from climate-driven shifts in human behaviors and economic activities. For example, increases in shipping, oil and gas exploration and fishing due to the loss of Arctic sea ice are highly likely to exacerbate acoustic disturbance, ship strikes, bycatch and prey depletion for Arctic cetaceans. In the tropics, climate change may result in increased hunting pressure on near-shore dolphins and whales off Asia, Latin America, Africa, and elsewhere as the availability of other marine resources diminishes. This study explores the range of potential consequences to cetaceans worldwide from predicted climate-driven shifts in human behavior, and evaluates the risks to particular species given their geographic ranges and habitat preferences. While concern about impacts of climate change on cetaceans has largely focused on polar species, the analysis presented here suggests tropical coastal and riverine cetaceans such as the Irawaddy dolphin, Indo-Pacific humpback dolphin, and finless porpoise are particularly vulnerable to those aspects of climate change that are mediated by changes in human behavior. Policy recommendations include the following: (1) information about cetacean populations should be incorporated into national, regional and international climate adaptation decisions wherever possible (for example, via GEF-sponsored adaptation initiatives); and (2) human-mediated impacts of climate change should be included in cetacean conservation and management plans, such as the management procedures of the International Whaling Commission (IWC), where possible. Because human responses to climate change are likely to evolve rapidly over the coming years and decades, it is important that local, regional and international cetacean conservation and management plans include regular reviews to allow them to adapt to new information.     

鲸豚声信号实时采集与分析系统

本文设计了一套基于FPGA的鲸豚声信号实时数据采集与分析系统,在采集存储水族馆内声信号的同时,应用改进的HHT算法处理鲸豚叫声中的click信号,实现实时的甄别瓶鼻海豚和白鲸功能,可用于针对大型哺乳动物的远洋实时跟踪科考。     

Short-term survival of severe propeller strike injuries and observations on wound progression in a bottlenose dolphin

Until recently, vessel collisions with small cetaceans were presumed rare, mainly as a consequence of limited reporting. Observations on dolphin wound healing from propeller strike injuries also remain scarce. We present an extreme case of a bottlenose dolphin (Tursiops truncatus) with multiple propeller wounds, including a penetration to the bone, where survival was possible for at least 23 days post injury. We used photographic records in conjunction with field observations to describe wound progression in the absence of treatment. Considering the severity of the wounds, it was surprising that the injuries were not immediately fatal. A practical solution remains to be found for the problem of odontocete vessel collisions, in particular for small highly mobile species using neritic waters. In view of the ongoing problem in the Hauraki Gulf and, indeed, worldwide, we recommend continued evaluation of cases and reporting of incidents, in addition to investigations into mechanisms that may reduce the risk of occurrence of vessel collisions with cetaceans.     

Towards best environmental practice for cetacean conservation in developing Scotland's marine renewable energy

Marine renewable energy is seen as an important component of the UK's future energy strategy and contribution to reducing the greenhouse gas emissions responsible for climate change. The UK aims to generate a total of 33 GW (gigawatts) of offshore wind energy. Its implementation strategy includes the development of ten offshore wind farms within Scottish territorial waters. In addition, between 1000 MW (megawatts) and 2600 MW of marine renewable energy generating capacity could be achieved in Scotland using wave and tidal power devices. However, there are negative environmental impacts associated with marine renewable energy. Intense noise is produced during pile driving, drilling and dredging operations with potential consequences for cetaceans. There are also increases in vessel activities during exploration, maintenance and construction with association risks of disturbance and collisions. Some underwater devices will be large and may be positioned in arrays across the habitats that cetaceans frequent. The consequences of encounters between cetaceans and such devices are as yet unknown. It is recommended that the Scottish Government complete full and transparent Marine Spatial Planning, including consideration of cumulative impacts, before moving to license appropriate sites.     

鲸的食性、摄食方式及其与渔业的关系

鲸具胎生、哺乳、用肺呼吸、体温恒定等典型哺乳动物的特征,由于终生生活在水中,体型庞大。要保持正常的体温和代谢活动。必须消耗大量食物来补充能量。因此,了解鲸的食性、摄食方式及其与渔业的关系就显得十分重要。按摄食方式的不同将须鲸划为过滤型,齿鲸为捕食型;按食物组成和摄食方法的不同,将齿鲸又分成三大类型：食乌贼型、食肉型和食鱼型;须鲸则为食浮游生物型。同时探讨了鲸类造成渔业的损失：直接摄食、毁坏渔具、妨碍作业、争食渔网内的渔获物、驱散鱼群造成鱼捕捞量降低等。渔业对鲸类的主要影响表现为：误捕、食物短缺和噪音等．     

Oceanographic influences on seabirds and cetaceans of the eastern tropical Pacific: A review

This paper is part of a comprehensive review of the oceanography of the eastern tropical Pacific, the oceanic region centered on the eastern Pacific warm pool, but also including the equatorial cold tongue and equatorial current system, and summarizes what is known about oceanographic influences on seabirds and cetaceans there. The eastern tropical Pacific supports on the order of 50 species of seabirds and 30 species of cetaceans as regular residents; these include four endemic species, the world’s largest populations for several others, three endemic sub-species, and a multi-species community that is relatively unique to this ecosystem. Three of the meso-scale physical features of the region are particularly significant to seabirds and cetaceans: the Costa Rica Dome for blue whales and short-beaked common dolphins, the Equatorial Front for planktivorous seabirds, and the countercurrent thermocline ridge for flocking seabirds that associate with mixed-species schools of spotted and spinner dolphins and yellowfin tuna. A few qualitative studies of meso- to macro-scale distribution patterns have indicated that some seabirds and cetaceans have species-specific preferences for surface currents. More common are associations with distinct water masses; these relationships have been quantified for a number of species using several different analytical methods. The mechanisms underlying tropical species–habitat relationships are not well understood, in contrast to a number of higher-latitude systems. This may be due to the fact that physical variables have been used as proxies for prey abundance and distribution in species–habitat research in the eastern tropical Pacific.Though seasonal and interannual patterns tend to be complex, species–habitat relationships appear to remain relatively stable over time, and distribution patterns co-vary with patterns of preferred habitat for a number of species. The interactions between seasonal and interannual variation in oceanographic conditions with seasonal patterns in the biology of seabirds and cetaceans may account for some of the complexity in species–habitat relationship patterns.Little work has been done to investigate effects of El Niño-Southern Oscillation cycles on cetaceans, and results of the few studies focusing on oceanic seabirds are complex and not easy to interpret. Although much has been made of the detrimental effects of El Niño events on apex predators, more research is needed to understand the magnitude, and even direction, of these effects on seabirds and cetaceans in space and time.     

Protecting small cetaceans from coastal development: Impact assessment and mitigation experience in Hong Kong

Since the early 1990s, there has been an active program in Hong Kong to manage and protect local populations of small cetaceans from the effects of massive development in the area. This paper reviews the progress that has been made. Only two species regularly occur there: the Indo-Pacific humpback dolphin and the finless porpoise. Because most development has occurred in the western waters of Hong Kong, where generally only the humpback dolphin occurs, most of the work has been conducted on that species. Development of large infrastructure projects (such as airports, bridges, expressways, power plants, fuel facilities, and container ports) in Hong Kong often results in land reclamation, dredging and dumping of spoils, pipe and cable laying, percussive and bored piling work, underwater blasting, large increases in vessel traffic, and other impacts. Several mitigation measures have been used with varying levels of success, including bubble curtains/jackets, exclusion zones, ramping up of piling hammers, acoustic decoupling of noisy equipment, vessel speed limits, no-dumping policies, and silt curtains. Baseline, construction-phase, and operational-phase cetacean monitoring is often conducted to evaluate the success of conservation measures put into place. The Environmental Impact Assessment process in Hong Kong has involved cetaceans to a degree perhaps higher than anywhere else in the world, and much can be learned from studying the successes and failures of this situation.     

Structure and mechanics of nonpiscine control surfaces

Animals display a variety of control surfaces that can be used for propulsion and maneuvering devises. For nonpiscine vertebrates, these control surfaces are primarily evolutionary modifications of the paired appendages (i.e., legs). The diversity of control surfaces can be classified with regard to the forces used for stability and maneuverability. For animals, the pertinent forces are pressure drag, acceleration reaction, and lift. These forces can be generated actively by motion of the control surfaces or passively from flows produced by movements of the body or external flow fields. Drag-based control surfaces are associated with paddling and rowing movements, where the limbs are oriented either in the vertical parasagittal plane or horizontal plane, respectively. The paddle is unstreamlined and has a triangular design with a broad distal end, thereby affecting a large mass of water. Appendages, which are used to generate lift-based forces, are relatively stiff hydrofoils. To maximize lift, the hydrofoil should have a crescent wing-like design with high aspect ratio. This shape provides the hydrofoil with a high lift-to-drag ratio and high propulsive efficiency. The tail flukes of cetaceans are streamlined control surfaces with a wing-like design. The flukes of cetaceans function in the hydrodynamic generation of forces for thrust, stability, and maneuverability. The three-dimensional geometry of flukes is associated with the production of lift and drag. Previous studies of fluke geometry have been limited in the number of species examined and the resolution of measurements.     

组织培养板在获取微藻种质中的应用

报道一种基于选择演替和及时显微跟踪的原理获取一次野外水样中多种微藻的方法。利用组织培养板方便的显微观测条件和适宜容积提供微藻存活的空间,在不同培养条件下,进行微藻群落演替、及时显微跟踪检测和优势种分离,可以使野外水样初期低生物量种类优势化,以利于纯系种质的获得。并通过不同演替条件获得其中出现的未鉴定或初期样品中未检出的种质,提供进一步研究的基础。该方法在获取微藻种质和种质库建设上不失为一可行技术手段。论文以一次野外采集海水样品在不同营养条件下的主要优势种类演替跟踪检测、低生物量种质优势化过程和纯系种质分离为例,说明该方法在获取微藻种质中的应用。     

免疫细胞化学法测定鱼类淋巴囊肿病毒(LCDV)滴度

利用免疫细胞化学法测定鱼类淋巴囊肿病毒(LCDV)滴度.以牙鲆鳃细胞系(FG)作为感染细胞,将生长旺盛的FG细胞接种于48孔培养板中培养至形成细胞单层,用2倍连续稀释的LCDV粗提液分别接种FG细胞.固定各稀释度LCDV感染后的FG细胞,孵育抗牙鲆LCDV单克隆抗体,其后再运用生物素-亲合素反应系统,以碱性磷酸酶底物APRed试剂盒发色.倒置显微镜观察,被病毒感染的FG细胞的细胞质呈现红色,未被感染细胞的细胞质呈无色.记录各稀释度病毒感染的阳性细胞孔数,按Reed-Muench法计算组织细胞培养半数感染量(TCID0).结果显示,免疫细胞化学法测得LCDV在FG的滴度为1.77×210 TCID50/mL.该法可以用来有效测定LCDV滴度,且结果直观、准确性较好,灵敏度较高.