European eels, Anguilla anguilla (L.), infected with Anguillicola crassus exhibit a more pronounced stress response to severe hypoxia than uninfected eels

The parasite, Anguillicola crassus is a non-native species that infects naive European eels, Anguilla anguilla, and causes pathological damage to the swimbladder, potentially compromising their ability to cope with hypoxic conditions. This study aimed to elucidate whether anguillicolosis exacerbates the stress responses to exposure to hypoxic water, conditions that have been implicated in mass mortalities of wild infected European eels. Blood parameters in infected and uninfected eels were measured during exposure to severe hypoxia over an 8-h period. Infected fish showed significantly higher levels of plasma cortisol compared with uninfected eels after 4 h of hypoxia. Uninfected fish showed an almost twofold increase in plasma glucose after 8-h exposure to hypoxia but infected fish showed no significant change, so that the plasma glucose concentration was significantly higher in uninfected eels than in infected eels. Both groups showed similar elevations in blood haematocrit, suggesting a similar catecholamine response in infected and uninfected eels. The lack of a hyperglycaemic response in infected eels, despite indirect evidence of a catecholamine response to hypoxia, may reflect an increase in glucose turnover. The data suggest that anguillicolosis results in a significantly greater corticosteroid stress response to hypoxia accompanied by a higher metabolic cost.     

Infection by Anguillicoloides crassus in a riverine stock of European eel,Anguilla anguilla

Abstract Infection by the parasitic nematode Anguillicoloides crassus Kuwahara, Niimi & Itagaki, in a wild riverine stock of European eel, Anguilla anguilla (L.), in a near pristine river was investigated. Samples were collected on a seasonal basis in distinct habitats along the river gradient. This study highlighted the presence of the parasite and completion of the whole life cycle in eels from the Río Esva. Infection levels by A. crassus were high at three sites between the mid river to the estuary and also varied among seasons. Condition of eels was lower at upstream sites compared with downstream locations. Although high‐quality, environmental conditions in the Río Esva may buffer the effects of A. crassus on eels, potential impacts and limiting factors for the parasite are discussed.     

The effect of foraging and ontogeny on the prevalence and intensity of the invasive parasite Anguillicola crassus in the European eel Anguilla anguilla

Infection patterns of the invasive Anguillicola crassus nematode were investigated in a population of the European eel Anguilla anguilla where parasite invasion is very recent, Loch Lomond, Scotland. Intensity levels of the parasite were associated with differences in fish ontogeny and trophic ecology. Although eels foraged on both fish and invertebrates, individuals which were smaller and fed on invertebrates (>70% contribution to diet) were found to contain a greater number of swim bladder parasites compared to larger eel with a predominance of fish (>60% contribution) in their diet. Within affected fish, a significant negative relationship was found between fish length and parasite intensity, with smaller individuals having higher parasite intensity than larger individuals. This study indicates that food intake and infection risk are linked in this recently infected host–parasite system. From a management perspective increasing our understanding of how infection intensity and repeated exposure is linked to resource use in an ecosystem is important for the future management of this endangered species in Europe.     

The pathogenic helminth parasites of eels

Although 63 and 55 species of helminths have been reported from each species of Atlantic eel and from 29 to 19 for each species of Pacific eel only the monogeneans Pseudodactylogyrus bini and P. anguillae and the nematode Anguillicola crassus, originally specific to species of Pacific eels, can be considered serious pathogens. None of the three are normally pathogenic to their preferred natural eel host species in the wild. Pseudodactylogyrus spp. only cause serious local gill damage when present on a host in large numbers under optimal conditions that facilitate transmission. This is the case in eel aquaculture, where infections can be controlled by drugs. Anguillicola crassus is only pathogenic to Anguilla anguilla and A. rostrata when Atlantic eels are introduced to the far east or when the parasites have been introduced to Europe. Here the parasite life cycle differs in that A. crassus can infect a wide range of intermediate hosts, employ paratenic hosts and survive as larvae for months in the swimbladder wall. This makes it an excellent colonizer. Its major pathogenic effects on eels result from haemorrhaging in, and thickening of, the swimbladder wall. It reduces the oxygen concentration in the swimbladder, reducing its ability to function as a hydrostatic organ, and increases the stress response of eels. In shallow lakes at warm temperatures this can result in mass mortalities. It is also feared that the parasite affects the ability of eels to migrate to the Sargasso Sea and so contributes to the decline in eel populations. Control by drug treatment is possible in culture, but not in the wild.     

Comparative imaging of European eels (Anguilla anguilla) for the evaluation of swimbladder nematode (Anguillicoloides crassus) infestation

This study compares diagnostic imaging tools in detecting the parasitic swimbladder nematode Anguillicoloides crassus in Anguilla anguilla (L.) and focuses on ultrasound in an attempt to develop a non‐destructive, field diagnostic test. Ultrasound use could allow the parasite to be diagnosed without decreasing the number of critically endangered European eels through post‐mortem. In the preliminary study, eels were examined with computed radiography, computed tomography, magnetic resonance imaging, 14 MHz high‐end ultrasound and 5 MHz low‐end portable ultrasound, and the results were compared with post‐mortem findings. This ultrasound scanning technique did not produce any promising results. A second batch of eels was examined using the same high‐end and low‐end ultrasounds, but employing a different scanning technique and comparing the results with post‐mortem. This second study, scanning along the midline from below, allowed for the detection of anomalies associated with moderately infected animals. None of the eels used in this study were severely infected; thus, no conclusions can be made regarding the use of ultrasound in those animals. Overall, it was found that none of the techniques were useful in diagnosing mildly infected individuals; therefore, no single diagnostic imaging tool is sensitive enough to replace post‐mortem for definite diagnosis.     

An evaluation of indices of gross pathology associated with the nematode Anguillicoloides crassus in eels

This study compares two alternative indices for quantifying the gross pathology of the swimbladder of eels, Anguilla anguilla (L.), infected with the nematode Anguillicoloides crassus. Two observers recorded twice the scores obtained by the two indices on the same set of 71 wild caught eels (from elver to silver eels, French Mediterranean lagoons). The Length Ratio Index (LRI), performed better than the Swimbladder Degenerative Index (SDI), in three of four predefined criteria of decision. First, the LRI better correlated with an estimate of the swimbladder volume reduction, a functional consequence of the infection (representativeness). Also, the LRI was less prone to subjectivity (inter-observer variability) and more precise (intra-observer variability), although less easy to generate (time needed for measurement/assessment). Using a sub-sample of 32 unaffected eels (showing minor if any swimbladder damage and no living worms at autopsy), we ascertained a linear relationship between the swimbladder length and the total body length, a prerequisite of isometric growth, to definitively accept the new ratio index as a valid alternative to the SDI. Also, because the LRI can be recorded on live specimens with radio-imagery (non-invasive method), we recommend its use, and provide a graph of correspondence between the SDI scores, the LRI scores and the estimated proportion of gas loss in the swimbladder.     

Host‐specificity of the endangered thick‐shelled river mussel (Unio crassus,Philipsson 1788) and implications for conservation

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Preslaughter electrical stunning of eels

The current procedures for slaughtering European eels (Anguilla anguilla) for food are very slow and cause suffering. Although there is little legislation for protecting the welfare of fish at slaughter, the legislation covering farmed mammals and birds at slaughter is well defined, requiring that these animals be rendered insensible immediately or without fear or pain prior to being killed. For many mammals and birds this can be achieved using an electrical stun, which is then followed by a procedure that actually kills them, such as exsanguination. This paper reports the investigation of the possibility of using electricity to stun eels, rendering them insensible to pain. Using 1 s duration alternating currents at 50 Hz applied directly across the head of the fish, it was shown that it was possible to stun the fish with currents of 0.1 A and above. Increasing the applied current increased the length of the period of the stun. When the duration of the application of the current was increased to 30 s it was found that the fish could be killed using currents between 0.50 A and 0.95 A. These results show that it is possible to use electricity to instantly stun eels and also to kill them by using longer duration currents. The use of preslaughter electrical stunning at slaughter could allow the welfare of these fish at slaughter to be improved greatly.     

Local adaptation studies and conservation: Parasite–host interactions between the endangered freshwater mussel Unio crassus and its host fish

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The phylogeny of Anguillicola crassus and A. globiceps based on partial 18S ribosomal RNA sequences

The parasitic nematodes Anguillicola crassus and A. globiceps infect the swimbladder of eels. To investigate the relationship of these species at the molecular level, the small subunit ribosomal RNA (18S rRNA) genes of A. crassus and A. globiceps were cloned and their nucleotide sequences determined. The 18S rRNA gene of A. crassus and A. globiceps were found to have similar lengths (886 and 888 base pairs, respectively) and demonstrated extensive similarity in their nucleotide sequences, showing 98.8% homology. However, the sequence homology of the 18S rRNA gene of A. crassus with that of other nematodes was found to be in the range of 72.7–89.0% homology, whilst that of A. globiceps was in the range of 73.1–89.4% homology. A phylogenetic reconstruction showed A. crassus to be closely related to A. globiceps. The analysis of nucleotide sequences of the 18S rRNA gene of fish parasites has proven useful in supporting species status and phylogenetic relationships.     

Anthropogenic spreading of anguillid herpesvirus 1 by stocking of infected farmed European eels,Anguilla anguilla (L.), in the Schlei fjord in northern Germany

The Schlei fjord in northern Germany is the recipient water of a comprehensive eel, Anguilla anguilla (L.), stocking programme. Since 2015, stocked eels become alizarin red S marked, but to date no control mechanism is implemented in this stock enhancement measure to prevent anthropogenic spreading of diseases. Consequentially, it was possible that farmed stocking cohorts of 2015 and 2016 (in total ca. 1040 kg) were subsequently tested positive for anguillid herpesvirus 1 (AngHV 1). For this study, 100 eels [total length (TL) 24.3–72.9 cm, age ca. 1–6 years] were caught in 2016 and investigated with regard to AngHV 1 infection, parasite load (Anguillicoloides crassus) and body conditions. 68% of the eels were found to be virus positive while larger specimens were more often infected. In addition, a fitted generalized linear model (area under the curve = 0.741) demonstrated that an increase in individual TL is accompanied with an increased risk of clinically relevant virus loads. Anguillicoloides crassus turned out to be an important stressor for eels, because parasite and virus load revealed a significant positive correlation. The results of this study evidently show the urgent need of a disease containment strategy for eel stocking programmes.     

Genetic structure of New Zealand eels Anguilla dieffenbachii and A. australis with allozyme markers

Abstract – Glass eels and adults of Anguilla australis and A. dieffenbachii were collected from rivers in the North and South Islands of New Zealand and tested for nine and ten allozyme loci, respectively. There was no heterogeneity among glass eel samples, but there was heterogeneity among adult samples within species. The results indicate that there may be single panmictic populations subject to sweepstake events or different selective pressures in the adult environments.     

Evidence of a local spawning site of marine eels along northeastern Japan, based on the distribution of small leptocephali

Evidence of recent spawning by several species of marine eels of the families Congridae, Ophichthidae, and Nettastomatidae was found over the continental shelf along the coast of northeastern Japan in October 2003. Sixty‐three leptocephali of at least three taxa that ranged from 3.8 to 12.3 mm total length were collected in a relatively small area, just to the south of Cape Shioya, primarily over the 100‐and 200‐m‐depth contours. The leptocephali of Gnathophis nystromi nystromi were the most abundant, followed by those of G. nystromi ginanago. This spawning area is located in an area that appeared to include water originating from the Tsugaru Warm Current and is further south than the typical latitude of the cold water of the Oyashio First Intrusion. These findings suggest that some species of marine eels may not migrate past the slope to spawn, and that the oceanographic features of the region may provide favorable conditions for the growth and retention of leptocephali in the area to the south of Cape Shioya.     

人工诱导花鳗鲡的精巢发育成熟及其精子的生物学特性

用肌肉注射HCG的处理方式（剂量为500U/㎏•体重,每周注射1次,注射时间为6周）诱导雄性花鳗鲡性腺发育成熟,成熟率达80.0%。对人工催熟花鳗鲡精子的生物学特性研究结果表明：花鳗鲡精子头部长径为3.81±0.69µm,短径为1.24±0.15µm;尾部长度为24.83±3.05µm;精液pH为7.3~7.5,精子密度为1.02×1010尾/ mL。精子的适宜盐度为15~20,其中盐度为15时,精子激活比率最高,快速运动时间以及精子的寿命最长。精子的pH适宜范围为6.0~8.0,pH值过高或过低都会影响精子的活力与寿命。另外,4 种金属离子(Mg2+、Ca2+、Na+和K+)对花鳗鲡精子活力与寿命的影响趋势基本一致,金属离子浓度过高或过低都会抑制精子的活力、缩短快速运动时间和寿命。而MgCl2、CaCl2、KCl、NaCl溶液浓度为0.4~0.6g/mL时,精子活力最好,最高激活比率为3级（41.0%~60.0%）。     

中国东南沿海日本鳗幼体的发育时相及其迁徙路径分析

对2008年1—3月采自中国广东至江苏沿海9个河口的534尾日本鳗鲡幼体鉴定发育时相,分析了其中104尾玻璃鳗的日龄、柳叶鳗龄和抵岸龄。结果显示,中国大陆东南沿海玻璃鳗的柳叶鳗龄平均为102(90~112)d,并呈现出地域性的从南至北逐步增大的趋势。Scheffe′s多重比较可将9个群体按柳叶鳗龄的大小归为3个组,南部的新会和汕头归为第1组;中间的福清、宁德、温州、台州归为第2组;北面的慈溪、九段沙和大丰归为第3组。同样,群体的R-聚类和个体的Q-聚类分析也显示出与上述分组相一致的结果。研究表明,从产卵场随北赤道洋流迁徙至菲律宾东北部并进入黑潮接近中国东南沿海的柳叶鳗,分成3个群体迁徙。变态最早的进入了南方的水域,抵达广东沿海;变态较早的进入了中部的东海水域,抵达闽浙沿海;变态最晚的则被送到长江口及其以北的沿海河口。结果还显示,最北部大丰的平均柳叶鳗龄[(106.00±3.22)d]比最南部新会的平均柳叶鳗龄[(91.87±1.36)d]大14.13 d,其与黑潮流速(约96 km/d)的乘积(1 350 km)同两地的距离大致相等,研究表明,我国沿海的玻璃鳗在柳叶鳗阶段是随着黑潮暖流自南向北被动地迁移。     

The use of fyke nets as a quantitative capture technique for freshwater eels (Anguilla spp.) in rivers

Abstract  Fyke netting is a convenient and effective technique for capturing freshwater eels, and catch-per-unit-effort is usually assumed to be an index of eel abundance. The present study investigated the potential of depletion fishing using baited fyke nets to obtain population estimates of longfin eels Anguilla dieffenbachii Gray, in a river in the South Island of New Zealand. The probability of capture (capture efficiency) of a single night's fishing increased with increasing size of eel, and ranged from 0.2 to 0.9 for eels <400 mm, to 0.7–0.9 for eels ≥500 mm. The capture efficiency of baited vs unbaited nets was assessed in a small stream that was electric fished after netting trials were completed. Baited nets proved to be an effective method of assessing abundance of longfin eels (>400 mm) but not shortfins ( A. australis Richardson); capture efficiency of the population of longfins (the sum of fyke caught and electric fished eels) was 0.4 for a single night's fyke netting, increasing to 0.8 over four nights. Comparable efficiencies for shortfins were <0.1 and 0.3 respectively. Unbaited nets were markedly less efficient for both species.     

Haematological responses in the European eel, Anguilla anguilla L., to sublethal infestation by Anguillicola crassus in a thermal effluent of the Swedish Baltic

Abstract. Haematological variations were studied in a European eel population, Anguilla anguilla L., naturally infested by the sanguivorous nematode Anguillicola crassus. This occurred off the Swedish Baltic coast in an area receiving heated cooling water from a nuclear power station. The erythrocyte count, haematocrit, leucocrit, haemoglobin concentration, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentrations, the various cell types in peripheral blood, serum iron concentration, the amount of total serum protein, and scrum protein fractions α ₁ , β₁ , α ₂, β ₂ and γ were all measured. Each variable was correlated to: (1) the intensity of infestation, i.e. number of parasites per host; and (2) a parasitization index, i.e. the weight of parasites per somatic weight of the host. Most variables showed no or only minor reactions to the infestation. However, a marked increase in the γ -fraction of scrum proteins, reduced lymphocyte numbers and increased granulocyte numbers are indicative of a humoral and cellular immune response.