Hosts and transmission of the crayfish plague pathogen Aphanomyces astaci: a review

The crayfish plague pathogen, Aphanomyces astaci Schikora, has become one of the most well‐studied pathogens of invertebrates. Since its introduction to Europe in the mid‐19th century, it has caused mass crayfish mortalities, resulting in drastic declines of local populations. In contrast, North American crayfish usually serve as latent carriers, although they may also be negatively affected by A. astaci infections under some circumstances. Recent research benefiting from molecular tools has improved our knowledge about various aspects of A. astaci biology. In this review, we summarize these advances, particularly with respect to the host range and transmission. We highlight several aspects that have recently received particular attention, in particular newly confirmed or suspected A. astaci hosts, latent A. astaci infections in populations of European crayfish, and the relationship between A. astaci genotype groups and host taxa.     

Procambarus clarkii Girard as a vector for the crayfish plague fungus, Aphanomyces astaci Schikora

Abstract. Procambarus clarkii Girard, a native freshwater crayfish species of Lousiana, USA, was found to harbour the crayfish plague fungus, Aphanomyces astaci Schikora, in its cuticle as a benign infection. Under certain conditions, P. clarkii dies as a result of this parasite, and the A. astaci infection then becomes acute and can be transmitted to Astacus astacus (L.).
Therefore, it is now shown that at least three different species of North American crayfish ( Pacifastacus leniusculus Dana, Orconectes limosus Raff, and Procambarus clarkii Girard) can carry the crayfish plague fungus, A. astaci , can transfer the disease to other crayfish species, and under certain circumstances can die of its own infection.     

Variation in the hyphal growth rate and the virulence of two genotypes of the crayfish plague organism Aphanomyces astaci

Crayfish plague, a devastating disease of freshwater crayfish, is caused by an oomycete organism, Aphanomyces astaci. Currently five genotypes of A. astaci are known, but variable features between the strains or genotypes have not been studied extensively. This study analysed 28 isolates of the As genotype and 25 isolates of the Ps1 genotype and reveals that the radial growth rate is significantly (P < 0.001) different between these two genotypes, although highly variable inside the genotype As. Two Ps1 genotype isolates and two As genotype isolates with different radial growth rates were tested in an infection trial. Clear differences were detected in the development of mortality in the test groups. The representatives of the Ps1 genotype caused total mortality within a short time span. The As genotype isolates were much less virulent. The slow‐growing As isolate showed higher virulence than the As isolate with a high growth capacity. Although slow growth could be one survival strategy of the pathogen, several other mechanisms are involved in the pathogenicity and warrant further studies.     

黄海绿潮浒苔(Enteromorpha prolifera)生活史的初步研究

实验室条件下对2008年6月黄海中南部海域暴发的绿潮浒苔(Enteromorpha prolifera)样品进行培养,观察记录绿潮浒苔生活史不同世代的生长发育情况.结果显示,成熟藻体释放出的雌、雄配子结合后固着,随后发育形成新个体;刚释放出的孢子具有聚集生长的趋势,随后发育形成具有假根和叶状体分化的新个体;在生长衰败期,部分藻体体细胞发生明显变化,细胞膨胀后发生分裂,发育形成新个体并能在死去的藻体上附着生长;培养的叶状体片段两端迅速愈合,并显示生长极性,两端分别形成叶状体和假根.后两种单性生殖方式目前尚未见公开报道.     

Growth and maturation of Pacific saury Cololabis saira under laboratory conditions

This study details the growth and maturation processes of Pacific saury Cololabis saira from eggs to first spawning under laboratory conditions. They were reared at 20°C, and fed almost to satiation every day. There was no significant difference in the knob length (KnL) between males and females, and so data for both sexes were combined in the following Gompertz growth formula: KnL _t  = 277.1 exp[?exp(?0.015 (t ? 83.8))]. The first spawning was observed 243 days after hatching (DAH). At the beginning of spawning, when the mean KnL was about 250 mm, only several hundred eggs were spawned at most. The spawning continued, and the number of spawned eggs increased notably after 260 DAH. Correlation and stepwise multiple regression analysis of the gonad somatic index (GSI) of Pacific saury versus KnL, CF, and DAH revealed that only DAH (R = 0.88 and 0.72 for males and females, respectively) was significantly correlated with GSI of Pacific saury (P < 0.001). This result suggests that DAH is one of the most influential factors for maturation in this species.     

Artemia salina as pabulum for growing Penaeids under laboratory conditions

Growing interest in Artemia salina Leach is supported by its high protein content, the relative ease of its mass production, and its incompletely utilized potential biomass which is found in saltpans all over the world.Experiments involving the catching of adult brine shrimps by means of a 500 μm nylon net located on the main outflow canal were carried out in Margherita di Savoia saltworks in southeastern Italy preliminarily in August 1980. Replicates during May–July 1981 showed it was possible to collect about 250 g min^?1 of adult Artemia. According to the figures of the samples taken, it is reasonable to assume a brine shrimp standing crop of at least 50 tonnes (wet weight) over a 1000 ha surface area. The Artemia collected in this way were tested for feeding Penaeids under laboratory conditions, as sole food and as supplement to a dry selfmade diet. Firstly Penaeid growth supported by mussels was tested in comparison with frozen Artemia diet. The latter gave the best results with 30% daily supply of shrimp body (wet weight). Moreover Artemia were tested as a balancing component of a dry diet to correct a deficit of aminoacids and supply missing highly unsaturated fatty acids (HUFA).With this mixed diet it was enough to supply A. salina 2 days per week to achieve promising results which are considered a good compromise between economics and the growth rate of Penaeus japonicus Bate. Consequently, it is suggested that intensive and/or extensive Penaeid cultures be planted near saltpans, utilizing brine shrimp biomass as a food source which could otherwise be wasted.     

The influence of haemocyte number on the resistance of the freshwater crayfish, Pacifastacus leniusculus Dana, to the parasitic fungus Aphanomyces astaci

Abstract. The number of circulating haemocytes in crayfish, Pacifastacus leniusculus Dana, was reduced after injection of yeast cell walls (Zymosan) into the animals. Crayfish harbouring the parasitic fungus A. astaci as a 'latent' infection in melanized areas in the cuticle died due to unrestricted growth of unmelanized hyphae when they were injected with Zymosan. The β–1,3–gIucan laminaran, introduced into the haemocoel of P. leniusculus harbouring A. astaci hyphae in the cuticle, also caused a decrease in the total haemocyte counts (THC), but these animals survived the treatment and no unmelanized hyphae of A, astaci were found. The recovery of THC was found to be more rapid after laminaran injection than after Zymosan injection, which could explain why laminaran treated crayfish retained their resistance and could combat the A. astaci hyphae in the cuticle. The importance of the number of haemocytes in defence and well-being of crayfish is discussed.     

Diel timing of molting and metamorphosis of Panulirus japonicus phyllosoma larvae under laboratory conditions

ABSTRACT:   The timing of molting and metamorphosis was monitored for Panulirus japonicus phyllosoma larvae cultured in the laboratory. Larvae cultured under natural light–dark cycle molted synchronously within approximately 1.0 h before and after sunrise, except for a short period after hatching; the timing of molting changed with time of sunrise. When three artificial light–dark cycles were introduced, larvae molted around the start of lighting, irrespective of the light–dark cycles. In the event of a sudden change in the start or end of lighting, molting was regulated by an endogenous rhythm. The start of lighting had a greater impact on the timing of molting than the end of lighting, suggesting that sunrise is probably the critical signal for phase-setting in molting rhythm. Metamorphosis to the puerulus stage occurred within 0.4 h before and 1.2 h after sunset under a natural light–dark cycle, and the timing of metamorphosis was also changed artificially by regulating the end of lighting.     

Larval molting and growth of the Japanese spiny lobster Panulirus japonicus under laboratory conditions

ABSTRACT:   Ten newly hatched phyllosoma of Panulirus japonicus were cultured individually to monitor body length ( BL ) and intermolt period, and 2000 were cultured in groups to sample specimens for measurement of body weight. Phyllosoma were fed with Artemia and mussel gonad; the culture seawater temperature was 24–26°C. The individually cultured phyllosoma showed an increment in body length by the first molt of approximately 0.5 mm, and the molt increment increased to approximately 1 mm at 5 mm BL ; it was constant to 15 mm BL . Thereafter, the molt increment increased exponentially. The duration of the first instar was 6–7 days. Instar duration increased with development up to approximately 2 weeks at the 20th instar (∼16 mm BL ) and then became constant. Of the 10 larvae reared individually, five metamorphosed to the puerulus stage. The entire phyllosoma life ranged from 245–326 days (mean 289.0 days), and the number of instars ranged from 22–29 (mean 26.2). Body length in the final instar ranged from 28.50–33.10 mm (mean 30.280 mm). For the phyllosoma cultured in groups, relationships between BL and wet/dry body weights ( WW / DW , mg) were expressed as exponential equations: WW  = 0.0686 BL ^2.2023 and DW  = 0.0209 BL ^2.1905.     

Mating and spawning of freshwater crayfish, Austropotamobius pallipes Lereboullet, under laboratory conditions

Abstract. The study includes mating and spawning data of freshwater crayfish, Austropotamobius pallipes Lereboullet, obtained in four consecutive years (1987–1990). A total number of 586 wild-caught female A. pallipes in the 21·65 mm to 45·60mm carapace length (CL) size were held under laboratory conditions at different dates in September–October. Initial densities at mating time were 21 and 24 crayfish/m² with a 1:2 male:female ratio. High percentages of mating were obtained (97·4–100%) except in 1989 (57·5%). In all years, most of the matings (90%) were concentrated in a period of 8–12 days with mean water temperatures between 12°C and 13·5°C. Spawning took place a few days after mating (minimum 2 days, maximum 14 days) at mean water temperatures between 8·2°C and 10·8°C. In the first 3 years, almost 100% of mated females spawned. However, in the fourth year (1990), when the mean size of crayfish was smaller, only 72·4% of mated females spawned. The mean number of pleopodal eggs was 64 (maximum 220, minimum 18). Egg diameter ranged between 2·30 and 3·25 mm (mean value 2·78 mm). Pleopodal egg number was positively correlated with carapace length ( r = 0·72).     

Observations on the development of Bunodera luciopercae (Müller, 1776) (Trematoda: Allocreadiidae) under field and laboratory conditions

Abstract. The intramolluscan development of Bunodera luciopercae was studied in laboratory infected pisidia maintained at three thermal regimes and compared with the seasonal development within a population of naturally infected first intermediate hosts at Llyn Tegid, Wales. At 4–8°C B. luciopercae did not develop beyond the sporocyst stage, although the development at 16–21°C W e s similar to that reported by previous workers. The early stages of the intramolluscan development of B. luciopercae at Llyn Tegid were similar to the observed development at 16–21°C. However, the results suggest that falling autumn-winter temperatures may initiate the production of immature cercariae within first generation rediae. Cercariae matured and were shed at Llyn Tegid during a short period between March and May. Live, juvenile B. luciopercae were artificially transferred from recently killed perch Perca fluviatilis , gill netted from Llyn Tegid, into live uninfected, laboratory maintained perch. The effect of temperature on the development of B. luciopercae in its definitive host was investigated at four thermal regimes. At 16–20°C development did not proceed normally and the infection had disappeared after 130 days. At 4–8°C and at natural environmental temperatures, gravid flukes were produced after 158–168 days. An increase in temperature on infected fish maintained at 4–8°C for 43 days did not stimulate parasite maturation. It is suggested that B. luciopercae may require an extended period of vernalization to elicit maturation in its definitive host and that temperature has a major influence on the life history of this parasite.     

Productivity of Sargassum linearifolium in potassium fortified inland saline water under laboratory conditions

Growing aquatic species in inland saline water (ISW) is one way to reduce the adverse impact of ISW to agriculture farms. This 84‐day laboratory‐trial was conducted to study the growth of Sargassum linearifolium cultured in ocean water (OW), ISW, ISW fortified with potassium equivalent to 100% (ISW100), 66% (ISW66) and 33% (ISW33) of potassium in OW at 35 g/L. The biomass and cumulative specific growth rate (SGR) of S. linearifolium increased significantly (p < .05) with increased potassium in ISW until 56 days and then declined. The ISW100 and OW resulted in similar growth patterns and yielded peak biomass at day 42, proving static biomass for the next 28 days before declining. The biomass of S. linearifolium cultured in ISW and ISW33 significantly (p < .05) decreased and was lower than in ISW100 and died after day 56. The SGR of S. linearifolium in OW, ISW100 and ISW66 levelled off and showed no difference during the first 56 days. The S. linearifolium biomass and SGR negatively and significantly (p < .05) correlated with the concentrations of nitrate, phosphate in all waters. The increased potassium concentration in ISW similar to its concentration in SW brought the growth of S. linearifolium cultured to a level that was similar in OW.     

实验室条件下鱼类视角与个体邻近距离关系的探究

在麦穗鱼（Pseudorasbora parva）鱼群中,随着邻居鱼的靠拢,其影像在本鱼眼睛中占的视角越大。为了进一步探究其他的鱼群是否也存在相同的规律,本实验选用个体体长较小的喜成群的红鼻剪刀鱼（Hemigrammus rbodostomus）[由30尾组成,平均体长：（20.3±0.2）mm]作为对比研究对象。将其放置在水深5cm的水族缸（78.0cm×52.0cm×65.0cm）中,使用摄像机从正上方拍摄鱼群影像。逐幅提取鱼群影像中各个体的头部、质心（近头部1/3处）、尾部的二维数据,经过数据处理,分析视角与NND间的关系。结果表明：1）红鼻剪刀鱼与麦穗鱼的视角均集中在80o内,即本鱼观察邻居鱼的视角不大于80度。2）在体长差异较大的两种鱼群中,邻近个体占据本鱼的视野范围不会因为鱼体的增大而增加。3）邻近个体占据本鱼视野的最大视角与两者NND间满足以下关系：xy=A,y为视角（用弧度表示）,x为NND（单位:BL）,A为常数。该规律适用于麦穗鱼与红鼻鱼群体。4）该关系式表示,当邻居鱼靠近本鱼时,为了避免碰撞,鱼类个体用视角作为控制的主要参数,调整相互距离;此表明视觉在起主导作用,而侧线,嗅觉、电感觉、磁感觉等即使起作用,也处于次要的地位。     

Predation by scavenging amphipods to injured hatchery-raised juvenile Japanese flounder Paralichthys olivaceus under laboratory conditions

ABSTRACT:   The attacking potential of the scavenging amphipod Scopelocheirus onagawae on artificially injured hatchery-raised Japanese flounder Paralichthys olivaceus juveniles was investigated in relation to the degree of injury on the fish. All injured flounder juveniles were attacked by amphipods regardless of the degree of injury, while non-injured juveniles were not attacked. The attack by amphipods on the juveniles generally depended on the amount of glycine, a main feeding stimulant for the amphipod, released from the injury opening. The swimming ability of flounder juveniles was important to avoid the attack of amphipods. Furthermore, an area of injury allowing the amphipods to cling to the fish affects to the vulnerability of juveniles against the predation of amphipods. This study suggests that scavenging amphipods are potentially involved in the rapid reduction of the number of hatchery-raised juveniles.     

Feeding selectivity of the seahorse,Hippocampus kuda (Bleeker), juveniles under laboratory conditions

This study examined the feeding selectivity of Hippocampus kuda juveniles under captive conditions and evaluates different food organisms that could be used to improve hatchery‐rearing of this species. Newly born H. kuda were reared for 10 days in 60‐L capacity tanks and fed rotifers (Brachionus rotundiformis), zooplankton (mostly Pseudodiaptomus annandalei and Acartia tsuensis) alone or both food sources. The size and amount of food ingested increased as seahorses grew. Selective feeding of seahorses appeared to change as they develop, preferring copepod adults over nauplii and rotifers. A. tsuensis was highly selected by juveniles over P. annandalei. Specific growth rate in terms of body weight (SGR‐BW, 15% day^–1) was the highest and mortality rate (9% at day 10) the lowest in seahorses fed a mixed food sources. Slowest growth rate (0.3% day^–1) and highest mortality rate (60% at day 7) were observed in seahorses fed rotifers alone. These results indicate that copepods are suitable food for seahorse juveniles, but a mixture of food organisms in the rearing tank environment enhances survivorship and growth of H. kuda, thus potentially providing a source of cultured rather than wild specimens for characterizing the life history of this threatened species.     

实验室条件下麦穗鱼群体动态结构的表达

为了探究麦穗鱼群体动态结构的特征参数,以及这些参数可能反映的行为机理,本实验使用2台摄像机从俯视和侧视2个方向同时拍摄由13尾麦穗鱼组成的群体,获取连续时间内麦穗鱼群体中各个体的三维位置数据,对个体间最近邻近距离、视角、转角变化量、个体游泳速度等参数进行分析。结果表明:麦穗鱼个体最近邻近距离多数处于0.5~2 BL,偏好的最近邻近距离为0.6~0.8 BL;麦穗鱼总是将最邻近的个体保持在本鱼80°视野范围内;避免碰撞时,个体鱼转角改变量为0~30°;在无人为干扰的自然环境下,个体鱼以0.75 BL/s左右的速度配合其他个体保持运动速度一致性。实验观察证实,视觉对麦穗鱼个体间的分布起着关键性作用,此外麦穗鱼群体动态结构还受到个体状态(饱食与饥饿)的影响。     

Effect of salinity and temperature on marine leech,Zeylanicobdella arugamensis (De Silva) under laboratory conditions

The high prevalence (80–100%) of the marine leech Zeylanicobdella arugamensis (De Silva) on cage‐cultured Asian sea bass Lates calcarifer (Bloch) led us to investigate the percentage of juvenile leeches hatched from deposited cocoons, survival of juvenile and adult marine leeches at different salinity and temperature. The results showed that the hatching percentage of juvenile leeches was highest at salinity of 30 ppt (32.5 ± 2.8%) followed by 20 ppt (18.0 ± 4.3%) and 10 ppt (12.1 ± 1.4%), respectively. It was found that the adult and juvenile leeches could live up to an average range of 4–7 days at salinity ranging from 10 to 40 ppt. The juvenile leeches were able to hatch at temperature ranging from 25 to 35 °C but unable to hatch at 40 °C. The survival period of adult and juvenile leeches ranged from 11 to 16 days at 25 °C, which was comparatively longer than 5–13 days and 10 h – 5 days at 27–30 °C and 35–40 °C, respectively. The study provided the information on the physical parameters of salinity and temperature which are most optimal for the marine leech Z. arugamensis to propagate.