Regeneration of the caudal and pectoral fins of a bony fish, Gambusia (Haplochilus) schoelleri

The study showed that excision of the caudal fin at basal level was followed by complete regeneration in one and a half months, whereas if it was amputated at mid-fin level, complete regeneration took two months. The findings confirm that the greater the extent of amputation, the faster the rate of regeneration. In the case of the pectoral fin, only part of which was removed, it was found that the fin completely regenerated, with recovery of its original pattern, within two months after amputation.     

Salinity tolerance of the mosquito fish, Gambusia affinis (Baird and Girard)

Experiments were conducted to determine the adaptability of mosquito fish to various salt concentrations. Mosquito fish, Gambusia affinis , (total length (T.L.) 27–37 mm) were subjected to abrupt and gradual changes from fresh water to various salinities (S). Low mortality (10%) occurred when the fish were transferred from fresh water (S = 0·5‰) to 50% sea water (S = 19·5‰ Through gradual adaptation 65·0% of the fish were able to tolerate 100% sea water for 7 days. After a 7 -day stay in sea water, fish were readapted to fresh water during a 3-h period. The results indicate that they were well able to tolerate the abrupt change. Through a gradual adaptation fish were also able to tolerate salinities ranging from 39·0‰ (100% sea water) to 58·8‰     

Genomics and mapping of teleostei (bony fish)

Until recently, the Human Genome Project held centre stage in the press releases concerning sequencing programmes. However, in October 2001, it was announced that the Japanese puffer fish (Takifugu rubripes, Fugu) was the second vertebrate organism to be sequenced to draft quality. Briefly, the spotlight was on fish genomes. There are currently two other fish species undergoing intensive sequencing, the green spotted puffer fish (Tetraodon nigroviridis) and the zebrafish (Danio rerio). But this trio are, in many ways, atypical representations of the current state of fish genomic research. The aim of this brief review is to demonstrate the complexity of fish as a group of vertebrates and to publicize the 'lesser-known' species, all of which have something to offer.     

Similarity in yolk-platelet structure of an ancient bony fish (Acipenser) and an ancient reptile (Sphenodon)

Lipovitellin-phosvitin crystals from oocytes of the sturgeon (Acipenser boeri L.) and the tuatara (Sphenodon punctatus [Gray]) have been shown to he orthorhombic like all such known crystals from vertebrates higher than cyclostomes. Lattice parameters and projected crystal views also resembled closely the hitherto known cases. The high evolutionary conservation of this crystal structure probably reflects a cell-biological significance of the tertiary structure of crystal constituents, especially of the lipovitellines. A comparative consideration of crystallographic data from cyclostomes and higher vertebrates suggests that cyclostomes started with one species and all higher vertebrates with two species of lipovitellin molecules.     

Molecular,cellular and histological changes in skin from a larval to an adult phenotype during bony fish metamorphosis

Developmental models for skin exist in terrestrial and amphibious vertebrates but there is a lack of information in aquatic vertebrates. We have analysed skin epidermal development of a bony fish (teleost), the most successful group of extant vertebrates. A specific epidermal type I keratin cDNA (hhKer1), which may be a bony-fish-specific adaptation associated with the divergence of skin development (scale formation) compared with other vertebrates, has been cloned and characterized. The expression of hhKer1 and collagen 1α1 in skin taken together with the presence or absence of keratin bundle-like structures have made it possible to distinguish between larval and adult epidermal cells during skin development. The use of a flatfish with a well-defined larval to juvenile transition as a model of skin development has revealed that epidermal larval basal cells differentiate directly to epidermal adult basal cells at the climax of metamorphosis. Moreover, hhKer1 expression is downregulated at the climax of metamorphosis and is inversely correlated with increasing thyroxin levels. We suggest that, whereas early mechanisms of skin development between aquatic and terrestrial vertebrates are conserved, later mechanisms diverge. This work was carried out within the project “Arrested development: The Molecular and Endocrine Basis of Flatfish Metamorphosis” (Q5RS-2002-01192) with financial support from the Commission of the European Communities. It does not necessarily reflect the Commission’s views and in no way anticipates its future policy in this area. This project was further supported by Pluriannual funding to CCMAR by the Portuguese Science and Technology Council. M.A. Campinho was sponsored by the Portuguese Ministry of Science (grant no. SFRH/BD/6133/2001).     

Development of a technique for quarantining Great Artesian Basin springs from colonisation by the invasive fish Eastern Gambusia (Gambusia holbrooki)

A species recovery project for the endangered fish Red‐finned Blue‐eye (Scaturiginichthys vermeilipinnis) commenced in 2009 at Edgbaston Reserve in central western Queensland. To date, the project has included relocations and control of alien Eastern Gambusia (henceforth Gambusia, Gambusia holbrooki), using the piscicide rotenone. Preventing Gambusia colonising springs where Red‐finned Blue‐eye occur or have been relocated is crucial to the success of the project. As such, this study tested the use of porous silt fences that were installed around five springs from 2012. Results indicate that silt fencing is a suitable barrier material and that provided installed silt fences are serviced regularly, the material is an effective method of quarantining populations of endangered species in isolated aquatic environments. Four of the five installed fences were effective, and maintenance consisted of small repairs at regular intervals (quarterly) followed by replacement after 2 years. As such, silt fencing may have wider application in the conservation of endangered fish and other organisms, particularly in areas where isolated waterbodies occur and where their size renders exclusion fencing a practical recovery action.     

Manual fish length measurement accuracy for adult river fish using an acoustic camera (DIDSON)

Total lengths (L_T) of 50 free-swimming fish in a tank, silver carp Hypophthalmichthys molitrix and rainbow trout Oncorhynchus mykiss, were measured using a DIDSON (Dual-frequency IDentification SONar) camera. Using Sound Metrics software, multiple measurements of each fish (L_T, side aspect angle and distance from the camera) at different times were analysed by two experienced operators while a subset of data was analysed by two inexperienced operators. The main result showed high variability in intra-fish L_T measurements. The number of measurements required to minimise errors and to obtain robust fish measurements (true L_T ± 3 cm) was estimated by a bootstrap method. Three to five measurements per fish were recommended for fish surveys in rivers. In this experimental study, aiming to reproduce river conditions, no evidence of fish position (side aspect angle and distance from the camera) effect was detected, but an operator effect (partially explained by training) was observed. General linear mixed models also showed that lengths of the smallest fish (L_T < 57 cm) were overestimated and lengths of the largest fish (L_T > 57 cm) were underestimated in comparison with their true lengths. In conclusion, we highlight that this technology, like any monitoring methods, returns imperfect observations. We advise DIDSON users to ensure that measurements are carried out correctly in order to draw accurate conclusion from this new technology.     

食蚊鱼（Gambusia spp.）入侵生态学研究进展

食蚊鱼(Gambusia spp.)是原产于北美洲的著名入侵物种,它对蚊类幼虫具有很强的捕食压力,被作为疟疾防治生物工具而在全世界温带和热带地区扩散.近20多年来的研究认为,食蚊鱼通过食物竞争和捕食等机制威胁引入地的无脊椎动物、鱼类、两栖类的生存,显著影响引入地的生物多样性.它们具有广泛的生境适应性、生长迅速、卵胎生、高生殖率、对生殖生态条件无特殊要求等特点.另外,它们的个体野外寿命不超过2a,种群更新速度快,种群内形成春季和夏季两个繁殖群体,并具有不同的繁殖生物学特点,因而形成复杂的世代结构.遗传上,雌鱼具有混交及能长期贮存精子的特点,能快速建立种群并克服奠基效应.这些种群通过快速适应性进化而形成一定规模的地理群体.当前,由于食蚊鱼在预防疟疾的工作中仍具有不可替代的作用,它会借助于人力的作用而继续扩散.为减轻它对非目标地区和非目标生物的影响,应进一步深入开展其入侵生态学研究.     

Parathyroid hormone-related protein (PTHrP) in cartilaginous and bony fish tissues.

Tissues from a range of fish were examined for the presence of parathyroid hormone-related protein (PTHrP) to investigate PTHrP protein distribution and PTHrP gene expression in jawless fish, cartilaginous fish, and bony fish. Immunoreactive PTHrP was localized using antisera to N-terminal and mid-molecule regions of human PTHrP and PTHrP gene expression examined using a digoxigenin labeled riboprobe to a conserved region of the mammalian PTHrP gene. In all of the fish studied, PTHrP protein and messenger RNA (mRNA) were localized to the skin, kidney, and skeletal muscle, following the pattern seen in higher vertebrates. Additional sites of localization for both protein and mRNA included gill, nerve cord, and pituitary, as well as developing dermal denticles and rectal gland in the elasmobranch species. The sites of PTHrP distribution indicate that PTHrP may have roles in ionoregulation as well as growth and differentiation in fish, as has been suggested in higher vertebrates. The results imply that the distribution of PTHrP is widespread in fish and that there is homology between the PTHrP molecules found in humans and fish. The conservation of localization and possible similarity of the PTHrP molecules between tetrapods and fish suggests that PTHrP has a number of fundamental roles in vertebrates. J. Exp. Zool. 284:541-548, 1999.     

Two divergent isotypes of the fourth complement component from a bony fish, the common carp (Cyprinus carpio)

Duplication and diversification of several complement components is a striking feature of bony fish complement systems. It gives an interesting insight into an evolutionary strategy for the possible enhancement of the repertoire of innate immunity. The present study is aimed at examining diversity in bony fish C4, a member of the thioester-containing complement components. Two diverged cDNA sequences sharing only approximately 32% identity at the amino acid level were isolated from the common carp and designated C4-1 and C4-2. C4-1 and C4-2 share a number of C4-like structural signatures, such as the thioester site and a disulfide-linked three-chain structure. Interestingly, they differ at the residue corresponding to the thioester-catalytic histidine, as seen in the human C4A and C4B isotypes, suggesting their distinct substrate specificities in the binding reaction of the thioester. Phylogenetic analysis indicates that the divergence of C4-1 and C4-2 predated the separation of the cartilaginous and bony fish lineages. Genomic Southern hybridization suggests the presence of single copy genes each encoding C4-1 and C4-2 in the carp genome. An activation fragment, C4a, was shown to be released from each isotype in carp serum activated via the classical and/or lectin pathways. Synthetic peptides representing a putative C2 binding site on C4-1 and C4-2 inhibited the classical pathway-mediated hemolytic activity of carp serum in a dose-dependent manner. The results suggest that C4-1 and C4-2 represent two major lineages of C4 that are present in carp serum, have distinct binding specificities, and are functional in the classical/lectin pathways of complement activation.     

The occipital region in the basal bony fish Erpetoichthys calabaricus (Actinopterygii: Cladistia)

Erpetoichthys calabaricus has unusual cranio‐vertebral anatomy, with an occipital centrum forming a component part of the compound basiexoccipital bone, and a ‘free‐floating’ occipital neural arch that differs from accessory arches found in some teleosts. The occipital neural arch bears autapomorphic lateral projections that articulate with small rod‐like bones resembling the spatial relationship of parapophyses and ribs, a feature normally restricted to vertebral centra. Based on analyses of cleared and stained specimens, computed tomography and histology, it is hypothesized that the lateral projections and associated rod‐shaped bones are structures that share developmental homologies to the unique ‘dorsal ribs’ of Polypteridae.     

Geographical and temporal variation of multiple paternity in invasive mosquitofish (Gambusia holbrooki,Gambusia affinis)

Multiple paternity (MP) increases offspring's genetic variability, which could be linked to invasive species' evolvability in novel distribution ranges. Shifts in MP can be adaptive, with greater MP in harsher/colder environments or towards the end of the reproductive season, but climate could also affect MP indirectly via its effect on reproductive life histories. We tested these hypotheses by genotyping N = 2,903 offspring from N = 306 broods of two closely related livebearing fishes, Gambusia holbrooki and Gambusia affinis. We sampled pregnant females across latitudinal gradients in their invasive ranges in Europe and China, and found more sires per brood and a greater reproductive skew towards northern sampling sites. Moreover, examining monthly sampling from two G. affinis populations, we found MP rates to vary across the reproductive season in a northern Chinese, but not in a southern Chinese population. While our results confirm an increase of MP in harsher/more unpredictable environments, path analysis indicated that, in both cases, the effects of climate are likely to be indirect, mediated by altered life histories. In both species, which rank amongst the 100 most invasive species worldwide, higher MP at the northern edge of their distribution probably increases their invasive potential and favours range expansions, especially in light of the predicted temperature increases due to global climate changes.     

Genetic variation at the Pgi locus in the mosquito fish Gambusia affinis (Poecilidae) and a possible effect on susceptibility to an insecticide

Although some studies have been completed on genetic variation in relation to tolerance to heavy metals, none have considered pesticide pollution. This study examines the relationship between genotypes at the glucose phosphate isomerase ( Pgi ) locus in Gambusia and susceptibility to the organophosphate insecticide chlorpyrifos. Laboratory toxicity tests show significant differences in genotype frequencies between susceptible and tolerant groups offish from one site but not at others. A field survey examined genotype frequencies and levels of pesticide in the fish at seven sites. No chlorpyrifos was found at any of the sites, but a number of organochlorines were recorded. Significant correlations were found between three organochlorines (DDT/DDD, DDE and lindane) and frequences of particular genotypes. There were also significant differences in tolerance between populations and between size classes. These results are discussed in relation to previous work with heavy metals.     

A new pseudophyllidean from a freshwater fish, Gambusia affinis, in China

Coelobothrium gambusiense n. sp. (Bothriocephalidae) was collected and described from the intestine of the freshwater fish Gambusia affinis (Baird and Girard) (Poeciliidae) in Fujian Province, Peoples' Republic of China. It is the first record of Coelobothrium in China. The parasite closely resembles Coelobothrium monodi Dollfus, 1970, from Capoeta damascina (Valenciennes, Cyprinidae) in Iran and Coelobothrium oitense Kugi and Matsuo, 1990, from Tribolodon hakonensis (GUnther, Cyprinidae) in Japan in general morphological characters, the scolex, and the incomplete proglottids. The third species of Coelobothrium is distinguished from its congeners by its much shorter strobila, presence of a neck, a bilobed ovary instead of a transversely elongated ovary, larger eggs, different final host and locality, and other morphological characters.     

Microsatellites from the world's heaviest bony fish,the giant Mola mola

We report the isolation and characterization of CA_n microsatellite loci from the Mola mola genome using a variation of the hybrid capture method. Five loci exhibit high levels of heterozygocity and two show significant sequence similarity to genes from the closely related Fugu rubripes. These microsatellite markers should be useful in multidisciplinary efforts to understand population characteristics of the giant ocean sunfish.     

Complement system of bony and cartilaginous fish

Accumulating evidence indicates that the complement system experienced a discontinuous development at an early stage of vertebrate evolution. Invertebrates such as echinoderms and ascidians, and the most primitive extant vertebrates, the cyclostomes, seem to have a primitive complement system equipped only with the alternative and lectin pathways. In contrast, cartilaginous fish and higher vertebrates seem to have a modern complement system which has two additional pathways, namely the classical and lytic pathways. Recent molecular analyses of the complement system of bony and cartilaginous fish have not only confirmed the above conclusion, but also revealed a unique characteristic of the complement system of fish, where certain key component genes are duplicated. The complement system seems to play a more pivotal role in body defence in fish, whose adaptive immunity is considered to be at a relatively undeveloped state.     

Investigating the morphology, function and genetics of cytotoxic cells in bony fish

Bony fish (teleosts) possess multiple cytotoxic cell lineages that recognize and destroy virally infected and transformed cells. In general, these lineages parallel their functional equivalents in mammals and include neutrophilic granulocytes, macrophages, cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. These four cell types have been morphologically identified in multiple fish species but only limited information is available about their function. In contrast, much work has gone into examining the function of a fifth cytotoxic cell lineage, termed nonspecific cytotoxic cells (NCC), that has been referred to as the bony fish equivalent of NK cells. However, evidence suggesting that NCC do not represent the NK lineage has come through the development of multiple cytotoxic catfish cell lines that are morphologically and functionally similar to human NK cells and are distinct from NCC. In addition to characterizing cytotoxic cells from fish, recent work has identified the novel immune-type receptors (NITR) and cichlid killer leukocyte receptors (cKLR) that are structurally related to mammalian NK receptors and likely play a role in cytotoxic function in fish. This review summarizes the morphological and functional evidence for cytotoxic cells within bony fish and discusses future directions for examining cytotoxicity through genomics and transgenics.