基于物理模型实验的光倒刺鲃生态行为学研究

建立了基于天然河道的物理模型,通过控制实验研究水环境因子对鱼类行为的影响。文中选取了金沙江下游2.6 km河段建立物理模型,以南方水系的经济鱼类光倒刺鲃为研究对象,进行了鱼的行为对底质和流速的响应实验。分析得出,光倒刺鲃对砂卵石底质的选择明显大于其它底质且差异极其显著(P<0.01),光倒刺鲃2龄幼鱼期的喜好流速范围为0.3-0.6 m/s;研究同时发现水流紊动强度对光倒刺鲃行为具有重要影响。该研究结果可以为光倒刺鲃栖息地模型提供参考。     

Expression profiles of seven glutathione S-transferase (GST) genes in cadmium-exposed river pufferfish (Takifugu obscurus)

Glutathione S-transferase (GST; EC 2.5.1.18) plays a critical role in detoxification pathways. In this study, we report cloning and expression of seven genes of the GST family of the pufferfish Takifugu obscurus together with mRNA tissue distribution pattern and time-course of expression in response to exposure to cadmium. At basal levels of tissue expression, GST-Mu is highly expressed in liver compared with other tissues. When fish were exposed to cadmium (5 mg/L for 96 h), expression of GST-MAPEG, GST-Mu, GST-Omega, and GST-Zeta was greatly increased, whereas GST-Alpha and GST-Kappa genes showed no significant response. These findings suggest that gene expression of a number of GST isoforms in T. obscurus is modulated in response to exposure to cadmium. We propose GST-Mu, GST-Theta, and GST-Zeta as candidate biomarkers for heavy metal exposure in this fish.     

Chemical and physical defenses against predators in Cystodytes (Ascidiacea)

Ascidians utilize both physical (spicules, tunic toughness) and chemical defenses (secondary metabolites, acidity) and suffer relatively little predation by generalist predators. The genus Cystodytes (Polycitoridae) is distributed widely in both tropical and temperate waters. Secondary metabolite composition, calcareous spicules and tunic acidity (pH < 1) may act as redundant defense mechanisms against predation in this genus. To assess the relative importance of chemical and physical defenses against predation in ascidians, we studied purple and blue morphs of Cystodytes from the western Mediterranean (formerly assigned to Cystodytes dellechiajei, but recently shown to belong to two different species), and a purple morph from Guam (USA), identified as Cystodytes violatinctus. Crude extracts, spicules, ascididemin (the major alkaloid of the blue morph) and acidity were used in feeding trials to evaluate chemical and physical defense mechanisms in Cystodytes spp. We performed feeding experiments in the field with a guild of generalist fish (mostly damselfish), and in the laboratory with a sea urchin and a puffer fish. Our results showed that all crude extracts and ascididemin significantly deterred fish predation, but not sea urchin predation. However, neither acidity alone nor spicules at natural concentrations deterred feeding. These results and other studies on sponges and gorgonians suggest that secondary metabolites are the primary means of defense against fish predators. Spicules and tunic acidity may perform other ecological roles and/or target certain specialist predators.     

Philomena oncorhynchi: Effect of hormones on maturation in anadromous sockeye, Oncorhynchus nerka

Adult sockeye salmon, Oncorhynchus nerka (Walbaum), treated with salmon pituitary extract in July survived for up to 47 days and developed nuptial colours in both male and female fish during this period, whereas no such characteristics appeared in untreated control fish. Furthermore, results showed that 20 of 50 adult female Philomena oncorhynchi recovered from the 15 treated fish contained fully developed tailed larvae in the uterus as compared with only one of 44 female P. oncorhynchi from 31 untreated control fish. None of 51 worms from 13 stilboestrol treated adult sockeye showed development in utero beyond an elongated embryo, nor did the host fish develop nuptial colours. P. oncorhynchi and its immature sockeye host (approx. 1 year prior to spawning) remained unaffected by any of three separate hormone treatment experiments using injection of salmon pituitary extract, injection of 17-β-estradiol, and stilboestrol mixed in the food.     

Micro-organisms in Trypanosoma cobitis

Bacteria-like micro-organisms have been found in the epimastigote and metacyclic trypomastigote stages of a fish trypanosome, Trypanosoma cobitis (Mitrophanow 1883), in both the leech vector, Hemiclepsis marginata and in culture, and also in the trypomastigote stage from the blood of infected fish.     

Identification of zoonotic Giardia genotypes in fish

Apart from a single record in a shark, there have been no published studies conducted on Giardia genotypes in fish. The present study investigated the prevalence of Giardia in cultured fingerlings (n = 227), wild freshwater (n = 227) and wild marine/estuarine species (n = 255) of fish in Western Australia by PCR amplification at the 18S rRNA, glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi) and beta-giardin (bg) loci. Results revealed a low prevalence of Giardia, 3.8% (27/709), in fish hosts. The zoonotic Giardia species, Giardia duodenalis assemblages A, B as well as G. duodenalis assemblage E and Giardia microti were detected. The identification of zoonotic species of Giardia highlights the public health importance of investigating parasites within fish host species.     

Preference for structured environment in zebrafish (Danio rerio) and checker barbs (Puntius oligolepis)

Information about the welfare and husbandry of pet and laboratory fish is scarce although millions of fish are sold in pet shops and used in laboratory research every year. Inadequate housing conditions can cause behavioural problems also in fish since they are complex animals with sophisticated behaviour. In this study, we investigated the influence of environmental complexity on compartment preference and behaviour in zebrafish (Danio rerio) and checker barbs (Puntius oligolepis). For the preference test, large aquaria were divided by two semi-transparent walls of Plexiglas into an empty compartment, a structured compartment enriched with plants and clay pots, and a smaller compartment in-between, where food was provided. For observation, the empty and structured compartments were divided into six zones of similar size by defining three vertical layers and two horizontal areas (back vs. front area). Seven groups of six to nine zebrafish and seven groups of seven or eight checker barbs were observed on four days each (within a time period of ten days) to assess compartment use and activity, and to assess behavioural diversity and use of zones within compartments. Both zebrafish and checker barbs showed a significant preference for the structured compartment. Nevertheless, in neither species did behavioural diversity differ between the empty and structured compartment. Zebrafish used all zones in both compartments to the same extent. Checker barbs, however, used the structured compartment more evenly than the empty compartment, where they mainly used the lower and middle zones. These results suggest that zebrafish and checker barbs have a preference for complex environments. Furthermore, they indicate that the behavioural and ecological needs of fish may vary depending on species, and recommendations for husbandry should be specified at species level.     

Expression of gonadotropin subunits in roach (Rutilus rutilus, Cyprinidae) infected with plerocercoids of the tapeworm Ligula intestinalis (Cestoda)

Plerocercoids of the tapeworm Ligula intestinalis (Cestoda: Bothriocephalidea) have been reported to inhibit gametogenesis of their intermediate fish hosts. However, mechanistic studies are rare and the proximate cues leading to impaired reproduction still remain unknown. In the present study we investigated the effects of infection by L. intestinalis on reproductive parameters of roach (Rutilus rutilus, Cyprinidae), a common fish host of this parasite. Field studies on roach demonstrated that in both genders infection prevented gonad development. As revealed by quantitative PCR, infection was accompanied by essentially lower pituitary expression of follicle-stimulating hormone β-subunit (FSHβ) and luteinizing hormone β-subunit (LHβ) mRNA compared with uninfected roach, providing clear evidence for gonadotropin-insufficiency as the cause of arrested gametogenesis. Under controlled laboratory conditions infected roach showed lower mRNA levels of FSHβ but not of LHβ, despite histology revealing similar gonad stages as in uninfected conspecifics. These findings indicate the involvement of FSH rather than LH in mediating effects of infection early during gonad development in roach. Moreover, the impact of L. intestinalis on reproductive parameters of roach appeared to be independent of the parasite burden. Together, these data provide valuable information on the role of FSH and LH as mediators of parasite-induced sterilization in a vertebrate and implicate the selective inhibition of host reproduction by L. intestinalis as a natural source of endocrine disruption in fish.     

Effect of dietary fish meal replacement by red algae, Gracilaria arcuata, on growth performance and body composition of Nile tilapia Oreochromis niloticus

A 12-week long feeding experiment was initiated to evaluate the effect of dietary supplementation of red algae, Gracilaria arcuata, on the growth performance, feed utilization and body composition of Nile tilapia Oreochromis niloticus (Linnaeus, 1758). The fish were fed with an algae-free control diet (C) and three experimental diets which replaced conventional fish meal with varying levels of dried G. arcuata (20%, 40% and 60%, represented as G20, G40 and G60, respectively). The growth parameters of final weight (FW), weight gain (WG), percentage of weight gain (WG%), daily growth rate (DGR) and specific growth rate (SGR) were significantly reduced (P < 0.05) at all levels of algae incorporation compared to the control diet. Moreover, the negative impact of Gracilaria meal on the growth performance of Nile tilapia increased as the proportion of algae in the diet increased, with fish on diet G20 exhibiting a significantly higher growth performance than the fish on either of the G40 and G60 diets. On the other hand, the feed utilization parameters feed conversion ratio (FCR) and protein efficiency ratio (PER) did not show significant differences between the fish in the control group and those on diet G20, although poorer FCR and PER outcomes were achieved in the case of fish on diet G60. The content of moisture, protein and ash in muscle and carcass increased as the proportion of Gracilaria meal in the diets increased, but the reverse was true for lipid level. These results indicate that incorporation of less than 20% red algae, Gracilaria arcuata, could be feasible in the diet of Nile tilapia and further studies are recommended to optimize the level of algae to improve growth performance.     

Effects of Quillaja saponins on growth,metabolism, egg production and muscle cholesterol in individually reared Nile tilapia (Oreochromis niloticus)

The effects of supplementation of a Quillaja saponin (QS) mixture in the diets of tilapia have been studied using a respirometer system that allowed feeding and continuous measurement of oxygen consumption of individual fish. Five fish each were given control diet (C group) and control diet supplemented with 150 mg kg^?1 (S150 group) or 300 mg kg^?1 (S300 group) QS. At the end of 14 weeks the weight gain of the S300 group was significantly higher than control (P<0.05) whereas that of the S150 group had an intermediate value. The S150 group had a higher growth rate (P=0.05) after the first 3 weeks of feeding with the experimental diets, compared to the other two groups. At the end of the experiment the S300 group had significantly higher (P<0.05) average values for energy retention, apparent lipid conversion, carcass fat, energy and significantly lower (P<0.05) average values for apparently unutilised energy and carcass ash content compared to the C group. The corresponding values of the S150 group were intermediate between the C and S300 groups. One out of two female fish in the S150 group and both female fish in the S300 group never produced eggs during the entire 14-week experimental period. Contrarily, all three female fish in the control group and one out of the two female fish in the S150 group regularly produced eggs, at a rate of approximately once in every 14 days. The muscle cholesterol level in the S300 group was significantly higher than that of the C group. Possible mechanisms of action of the dietary saponins are discussed.     

Novel Myxobolus and Ellipsomyxa species (Cnidaria: Myxozoa) parasiting Brachyplatystoma rousseauxii (Siluriformes: Pimelodidae) in the Amazon basin,Brazil

We describe two novel myxosporean parasites from Brachyplatystoma rousseauxii, an economically important freshwater catfish from the Amazon basin, Brazil. Myxobolus tapajosi n. sp., was found in the gill filaments of 23.5% of 17 fish, with myxospores round to oval in frontal view and biconvex in lateral view: length 15 (13.5–17) μm and width 10.7 (9.6–11.4) μm; polar capsules equal, length 5.8 (4.6–7.1) μm and width 3 (2.3–3.8) μm containing polar tubules with 6–7 turns. Ellipsomyxa amazonensis n. sp. myxospores were found floating freely or inside plasmodia in the gall bladder of 23.5% of fish. The myxospores were ellipsoidal with rounded extremities: length 12.8 (12.3–13.6) μm and width 7.6 (6.7–8.7) μm; with two equal, slightly pyriform polar capsules, length 3.8 (3.8–4.0) μm and width 3.1 (2.5–3.4) μm, containing polar tubules with 2–3 turns. We combined spore morphometry, small-subunit ribosomal DNA data, specific host, and phylogenetic analyses, to identify both of these parasites as new myxozoan species. Maximum likelihood and Bayesian analyses showed that Myxobolus tapajosi n. sp. clustered in a basal branch in a subclade of parasites from exclusively South American pimelodid fishes. Ellipsomyxa amazonensis n. sp. clustered within the marine Ellipsomyxa lineage, but we suspect that although the parasite was collected in freshwater, its hosts perform a large migration throughout the Amazon basin and may have become infected from a brackish/marine polychaete host during the estuary phase of its life.     

Disruption of pituitary-ovarian axis by carbofuran in catfish, Heteropneustes fossilis (Bloch)

We investigated whether carbofuran (CF), a carbamate pesticide, at sub-lethal concentration had any adverse effects on reproductive function of the Indian catfish, Heteropneustes fossilis. 17β-Estradiol content of serum and ovary of pre-spawning (P) and spawning (S) fish was reduced after sub-lethal concentration of carbofuran treatment (0.5–2 mg/ml, 30 days). After 30 days of CF treatment, the serum and ovarian vitellogenin levels of fish at the P stage were also reduced but remained unaltered in the S stage. The staining intensity of the pituitary gonadotrophs of the pre-spawning fish was significantly higher in CF-treated fish compared to controls suggesting the inability of the pituitary gonadotrophs to release gonadotropin following CF treatment. CF thus acts as an antiestrogenic, endocrine-disrupting agent in fish, possibly targeting the pituitary-gonad axis.     

Disparate infection patterns of Ceratomyxa shasta (Myxozoa) in rainbow trout (Oncorhynchus mykiss) and Chinook salmon (Oncorhynchus tshawytscha) correlate with internal transcribed spacer-1 sequence variation in the parasite

Ceratomyxa shasta is a virulent myxosporean parasite of salmon and trout in the Pacific Northwest of North America. The parasite is endemic in the Klamath River, Oregon/California, where a series of dams prevent movement of fish hosts between the upper and lower parts of the basin. Ceratomyxa shasta exhibits a range of infection patterns in different fish species above and below the dams. We hypothesised that the variations in infection and disease are indicators that different strains of the parasite exist, each with distinct host associations. Accordingly, we sought to identify strain-specific genetic markers in the ssrRNA and internal transcribed spacer region 1 (ITS-1). We examined 46 C. shasta isolates from water samples and two fish hosts, from June 2007 field exposures at upper and lower Klamath River sites with similarly high parasite densities. We found 100% of non-native rainbow trout became infected and died at both locations. In contrast, mortality in native Chinook salmon was <10% in the upper basin, compared with up to 40% in the lower basin. Parasite ssrRNA sequences were identical from all fish. However, ITS-1 sequences contained multiple polymorphic loci and a trinucleotide repeat (ATC)_0-3 from which we defined four genotypes: 0, I, II and III. Non-native rainbow trout at both sites were infected with genotype II and with a low level of genotype III. Chinook salmon in the upper basin had genotypes II and III, whereas in the lower basin genotype I predominated. Genotype I was not detected in water from the upper basin, a finding consistent with the lack of anadromous Chinook salmon there. Genotype O was only detected in water from the upper basin. Resolution of C. shasta into sympatric, host-specific genotypes has implications for taxonomy, monitoring and management of this significant parasite.     

Effects of the eyefluke, Diplostomum spathaceum, on the behaviour of dace (Leuciscus leuciscus)

The behaviour of dace infected with Diplostomum spathaceum was investigated in the laboratory. As the number of parasites present in the eye increased, the efficiency with which the fish fed on Gammarus pulex declined. The loss of efficiency was compensated for by an increase in the time devoted to feeding. Heavily infected fish spent more time in the surface layers of the water. This may increase the likelihood that a gull might eat the fish and thereby continue the life-cycle of the parasite. The parasite modifies the behaviour of its host in a way that increases the parasite's chances of survival at the expense of the host.     

Identification of Mediterranean Diplodus spp. and Dentex dentex (Sparidae) by means of DNA Inter-Simple Sequence Repeat (ISSR) markers

The aim of this paper was to test the effectiveness of Inter-Simple Sequence Repeat (ISSR) technique in detecting specific molecular markers to genetically discriminate some species of teleost fish. As model species, we used four Mediterranean sea bream species (i.e. Diplodus annularis, D. puntazzo, D. sargus sargus, and D. vulgaris), and the common Mediterranean dentex Dentex dentex. Furthermore, the occurrence in our samples of two partially albino sparids (suspected, on a morphological basis, to probably belong to D. sargus sargus and to D. dentex, respectively) allowed ISSRs to be tested on doubtful specimens. Eight ISSR primers produced 95 out of 97 scorable polymorphic loci with a large number (63) of species-private bands. Maximum parsimony analysis, ΔK statistics, and assignment test were consistent in separating the five species and solving the taxonomic position of specimens of dubious attribution. The results obtained evidence the usefulness of ISSR markers in specimen and species identification, particularly when the morphological traits of fish make the classification problematic.     

Predator-prey interaction between hatchery-reared Japanese flounder juvenile, Paralichthysolivaceus, and sandy shore crab, Matutalunaris: daily rhythms, anti-predator conditioning and starvation

Predator-prey interaction between sandy shore crab, Matuta lunaris (Forskål, 1775), and juvenile Japanese flounder, Paralichthys olivaceus (Temminck et Schlegel), was investigated under controlled laboratory conditions. Possibility of training and conditioning hatchery-reared flounder to avoid predators was also examined. Crabs took over 75% of their daily ration at night when they were given access to prey 24 h a day. Large (64.8±5.4 g)- and medium (30.68±3.33 g)-sized crabs ate ca. 5.5±1.45 and 3.9±1.99 individuals of flounder (TL=4.96±0.23 cm) a day, respectively. When flounder juveniles that have experienced predation pressure by crabs encountered predators again, they exhibited better survival compared to the naive fish. Flounder juveniles were also conditioned either using small and, thus, benign predators, or large crabs over fence. The conditioned fish with either method were better able to avoid capture by crabs than naive fish, revealing that learning process should play an important role in their predator avoidance. Anti-predator performance was also compared between starved and fed flounder juveniles. Fed fish were rarely eaten by predators after 3 h of exposure, whereas starved fish continued to be eaten. Our results suggest that stock-enhancement program of Japanese flounder can be improved by applying proper feeding protocol and conditioning to avoid predators prior to release. Present research supports the idea that behavioural and ecological consideration for the target species is indispensable for the success of stock enhancement.     

Parameters of rewards on choice behavior in Siamese fighting fish (Betta splendens)

Five experiments were conducted with Siamese fighting fish (Betta splendens) to investigate how choices in a T-maze were affected by parameters of a social reward (aggression display to another male): presence or absence, amount, delay and distance traveled. Bettas showed a preference for the side associated with the presence of another male rather than the side associated with nothing (Exp 1), a greater length of time of the reward (Exp 2) and shorter delay (Exp 3). The animals were indifferent when one side offered a longer delay to a longer reward time compared with a shorter delay to a shorter reward time (Exp 4). What was most surprising, however, was that fish preferred to choose the side that was associated with swimming a greater distance to reach an opponent male (Exp 5). These experiments demonstrate that, while some parameters of a visual reward affect behavior in predicable ways (greater amount, shorter delay), the complex motivations underlying inter-male aggression can produce what appear to be paradoxical results.