The morphology of the chondrocranium of Mastacembelus armatus (Cuv. and Val.).
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Vanadium is widely distributed, occurring in many types of minerals, coal, and petroleum. Anthropogenic sources of vanadium originate from the production, processing, and wastes of these materials. The aquatic toxicity of vanadium to fish species is not well characterized. This study focused on the three-spined stickleback, Gasterosteus aculeatus, a small and widely distributed euryhaline species of fish. The three-spined stickleback is used as an effluent-monitoring species in both Canada and the United States. Five 96-h static renewal acute toxicity tests were performed in moderately hard water with adult fish. The geometric mean and range of the five 96-h LC(50)s based on measured concentrations of total vanadium in the test solution were 3.17 and 2.35-4.07 mg V/L, respectively. A conservative estimation of a safe concentration of vanadium that would not affect survival of adult three-spined sticklebacks over a 96-h exposure period in moderately hard water is approximately 0.30 mg V/L. A comparison with other fish species previously tested suggests that the three-spined stickleback is intermediate in sensitivity to vanadium. Information reported from this study may be useful in effluent toxicity identification evaluations and ecological risk assessments related to vanadium.
This study focuses on characterizing the acute toxicity of copper in freshwater to the threespine stickleback, Gasterosteus aculeatus, a small and widely distributed euryhaline fish. The threespine stickleback is used as an effluent monitoring species in both Canada and the United States, yet in some locations natural populations are listed as threatened or endangered. Four 96-h static renewal acute toxicity tests were performed in moderately hard water using U.S. EPA methods with adult fish (mean wet weight = 0.41 g/fish). The geometric mean of the 24-, 48-, 72- and 96-h LC(50)s based on measured concentrations of total copper (estimated dissolved copper in parentheses) in the test solutions were 382.2 (366.9), 278.7 (267.6), 256.6 (246.3), and 227.2 (218.1) microg Cu/L, respectively. Conservative estimates of acute toxicity thresholds, made using LC1 values, for adult threespine sticklebacks over 24-, 48-, 72- or 96-h exposure periods in moderately hard water are approximately 114.3 (109.7), 78.3 (75.2), 67.0 (64.3), and 52.4 (50.3) microg Cu/L, respectively. Test results were normalized to a range of water hardness from very soft to very hard using two U.S. EPA methods, the water hardness and the Biotic Ligand Model normalization procedures. Subsequently, interspecies sensitivity comparisons were made with aquatic animal species used in both the current and proposed U.S. EPA copper water quality criteria documents. Information reported in this study may be useful in effluent toxicity identification evaluations, ecological risk assessments and criteria development where copper is a concern.
In this laboratory experiment, the effects on fish behavior caused by butyl benzyl phthalate (BBP) were of interest. We showed that shoaling behavior and bottom-dwelling behavior in threespine stickleback, Gasterosteus aculeatus, were altered as a result of exposure to 0.1 mg/L BBP. Threespine sticklebacks, collected from a freshwater population in central Norway, were exposed to BBP for 26 days. BBP was administered daily through the water. We found that exposed fish aggregated more into one single shoal than control fish. Further, the exposed fish spent more time at the bottom of the test aquarium than the control fish. From these results we conclude that the behavior traits aggregation and bottom-dwelling activity may be suitable and sensitive in detecting effects of BBP in threespine stickleback.
In past years we have witnessed decreased reproductive capacity in many wildlife species due to exposure to chemicals with endocrine-disrupting properties. In this laboratory experiment male threespine sticklebacks Gasterosteus aculeatus exposed to 17 beta-estradiol (2.0 micrograms/g) dispersed in peanut oil, on days 1, 7, 14, and 21, showed impaired paternal care compared to control fish that were exposed to peanut oil only. There were no differences between the two groups in number of males that built nests or in courtship displays. However, exposed males started nest building significantly later than control males. This study suggests that some but not all essential traits of male reproductive behavior may be altered as a result of exposure to 17 beta-estradiol. To reveal harmful effects of chemicals with suggested reproductive-disrupting properties it is thus important to take a wide variety of variables related to reproductive behavior into consideration.
Circulating 11-ketotestosterone (11KT) levels are higher during the courtship phase than during the later parental phase in a number of male teleosts. The present study describes the temporal changes in 11KT levels and their relationships to changes in courtship behavior, after different number of spawnings, over the nesting cycle of the male three-spined stickleback, Gasterosteus aculeatus, a small teleost showing pronounced paternal behavior. Plasma 11KT levels, measured by radioimmunoassay, were approximately 34 times higher during the initial courtship phase than at the end of the following parental phase in spawned males. In addition, males that spawned with three or more females on a single day showed an earlier decline in 11KT levels and in courtship behavior compared to males that were only allowed to spawn with a single female. Plasma 11KT levels remained high in males not allowed to spawn.
Courtship declines and ceases while parental care increases in the presence of developing eggs during the nesting cycle of the male three-spined stickleback, Gasterosteus aculeatus. Furthermore, circulating 11-ketotestosterone (11KT) levels are higher during the initial "courtship phase" than during the later "parental phase," similar to that found in other paternal fishes. This study aimed to investigate a possible functional relationship between changes in 11KT levels and changes in reproductive behavior during the nesting cycle. To this end, groups of nonspawned and spawned male sticklebacks were sham-operated, castrated, or castrated and treated with 11-ketoandrostenedione (11KA), and the effects of the treatments on courtship and parental care were studied. Castration removed circulating 11KT, while 11KA replacement prevented the natural decline in 11KT during the parental phase (11KA converts to 11KT extratesticularly), as assessed by radioimmunoassay. Regardless of treatment, parental care remained low and courtship was present in all nonspawned males, even at the end of the experiment. However, courtship did eventually decline in castrated nonspawned males compared to the other two nonspawned groups. In all treatments of spawned males there was a drastic decline in courtship and an increase in parental care. In castrated spawned males, however, the decline in courtship came earlier than in the other two spawned groups. 11KA treatment did not prevent the natural decline in courtship/increase in parental care in spawned males, indicating that the natural decline in 11KT is not responsible for the main portion of the rapid changes in these behaviors over the stickleback's nesting cycle. The limited effects of castration also exclude other gonadal hormones from being responsible for most of these changes.
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Vitellogenin (VTG) is a protein produced in the liver of oviparous animals in response to oestrogens. Abnormal production of VTG by males, therefore, is used as a biological indicator of exposure to xeno-oestrogens. In this study, a sandwich-ELISA for measuring VTG in Liza aurata (golden grey mullet) was developed and validated. Plasma VTG was purified from 17beta-oestradiol-injected immature individuals of mullet, by size-exclusion and ion-exchange chromatography. Polyclonal antibodies against VTG were raised in rabbits. A sensitive immunoassay was developed for measurement of vitellogenin in L. aurata serum, reaching a quantification limit of 0.01 microg mL(-1) and a dynamic range from 0.02 to 2 microg mL(-1). The assay is specific, because high levels (>100 microg mL(-1)) of carp (Cyprinus carpio), goldfish (Carassius auratus), tilapia (Oreochromis niloticus), tench (Tinca tinca), rainbow trout (Oncorhynchus mykiss), European eel (Anguilla anguilla) and frog (Rana perezi) purified VTG, give negligible responses. The assay was used to analyse plasma samples from wild mullet.
A quantitative real-time reverse transcription polymerase chain reaction (Q- RT-PCR) assay was developed for quantification of vitellogenin (Vtg) mRNA normalized to beta-actin in so-iuy mullet. Vtg mRNA in liver samples of so-iuy mullet was induced after a single injection of E2 (0.01, 0.1, 1.0 microg/g body) and a dose-response relationship was obtained. This method was applied to determine Vtg mRNA in so-iuy mullet collected from Liaodong Bay, Bohai Bay, NanDaiHe, and a control site in north China. Compared to the control, a high level of Vtg mRNA expression was detected in so-iuy mullets collected from NanDaiHe, whereas no obvious difference between Vtg mRNA expression from Liaodong Bay and Bohai Bay was found. Thus, this method is expected to be useful for further studying the potential of Vtg mRNA as a biomarker for assessing estrogenic activity in marine environments using the so-iuy mullet as a bioindicator species.
The mutational mechanism underlying the striking diversity in MHC (major histocompatibility complex) genes in vertebrates is still controversial. In order to evaluate the role of inter- and intragenic recombination in MHC gene diversification, we examined patterns of nucleotide polymorphism across an exon/intron boundary in a sample of 31 MHC class IIB sequences of three-spined stickleback (Gasterosteus aculeatus). MHC class IIB genes of G. aculeatus were previously shown to be under diversifying (positive) selection in mate choice and pathogen selection experiments. Based on recoding of alignment gaps, complete intron 2 sequences were grouped into three clusters using maximum-parsimony analysis. Two of these groups had >90% bootstrap support and were tentatively assigned single locus status. Intron nucleotide diversity within and among loci was low (p-distance within and among groups = 0.016 and 0.019, respectively) and fourfold lower than the rate of silent mutations in exon 2, suggesting that noncoding regions are homogenized by frequent interlocus recombination. A substitution analysis using GENECONV: revealed as many intergenic conversion events as intragenic ones. Recombination between loci may explain the occurrence of sequence variants that are particularly divergent, as is the case in three-spined stickleback, with nucleotide diversity attaining dN = 0.39 (peptide-binding residues only). For both MHC class II loci we also estimated the amount of intragenic recombination as population rate (4N(e)r) under the coalescent and found it to be approximately three times higher compared to point mutations (Watterson estimate per gene, 4N(e)mu). Nonindependence of molecular evolution across loci and frequent recombination suggest that MHC class II genes of bony fish may follow different evolutionary dynamics than those of mammals. Our finding of widespread recombination suggests that phylogenies of MHC genes should not be based on coding segments but rather on noncoding introns.