Snakehead Fishes Research Articles

Zoonotic human liver flukes, a type 1 biocarcinogen, in freshwater fishes: genetic analysis and confirmation of molluscan vectors and reservoir hosts in Bangladesh

BackgroundOpisthorchiid flukes, particularly Opisthorchis viverrini, Opisthorchis felineus, Clonorchis sinensis, and Metorchis spp. are the most common fish-borne zoonotic human liver flukes (hLFs). Liver fluke infections are more prevalent in resource-deprived and underprivileged areas. We herein estimated the prevalence of the metacercariae (MC) of major hLFs in common large freshwater fishes (lFWF) marketed for human consumption from some selected areas of Bangladesh along with detection of their molluscan vectors and reservoirs.MethodsThe current status of fish-borne zoonotic hLF infections in lFWF was investigated along with their molluscan vectors and mammalian reservoir hosts in Mymensingh and Kishoreganj in Bangladesh from July 2018–June 2022 using conventional and multiple molecular techniques, such as PCR, PCR-restriction fragment length polymorphism (RFLP), sequencing, and bioinformatic analyses. The infection rate of fishes was analyzed using the Z-test and the loads of MC were compared using the chi-squared (χ2) test.ResultsThe MC of C. sinensis, Opisthorchis spp., and Metorchis spp. were detected in 11 species of common and popular lFWF. In lFWF, the estimated prevalence was 18.7% and the mean load was 137.4 ± 149.8 MC per 100 g of fish. The prevalence was the highest (P < 0.05) in spotted snakehead fishes (Channa punctata, 63.6%). The highest rate of infection (P < 0.05) was observed with the MC of C. sinensis (11.8%). Metacercariae were almost equally (P > 0.05) distributed between the head and body of fishes. The infection rate was slightly higher in cultured (19.6%) fishes. The MC of C. sinensis, O. felineus, O. viverrini, and Metorchis orientalis in fishes were confirmed using PCR, PCR-RFLP and bioinformatics. The cercariae of opisthorchiid (Pleurolophocercus cercariae) flukes were only recovered from Bithynia spp. (3.9%, 42 out of 1089). The ova of hLFs from dogs (4.3%, 5 out of 116) and cats (6.0%, 6 out of 100), and adult flukes (M. orientalis) from ducks (41.1% 113 out of 275) were detected.ConclusionsThe MC of hLFs are highly prevalent in fresh water fishes in Bangladesh. Reservoir hosts, such as street dogs, cats, and ducks carried the patent infection, and residents of Bangladesh are at risk.Graphical

Chromosome complements of Channalucius and C.striata from Phu Quoc Island and karyotypic evolution in snakehead fishes (Actinopterygii, Channidae).

Snakehead fishes of the family Channidae are obligatory air-breathers freshwater predators, the vast majority of which belong to the genus Channa Scopoli, 1777. Channa species are characterized by high karyotypic diversity due to various types of chromosomal rearrangements. It is assumed that, in addition to the lifestyle, fragmentation and isolation of snakehead populations contribute to an increase in karyotypic diversity. However, the chromosome complements of many isolated populations of widespread Channa species remain unknown, and the direction of karyotype transformations is poorly understood. This paper describes the previously unstudied karyotypes of Channalucius (Cuvier, 1831) and C.striata (Bloch, 1793) from Phu Quoc Island and analyzes the trends of karyotypic evolution in the genus Channa. In C.lucius, the karyotypes are differed in the number of chromosome arms (2n = 48, NF = 50 and 51), while in C.striata, the karyotypes are differed in the diploid chromosome number (2n = 44 and 43, NF = 48). A comparative cytogenetic analysis showed that the main trend of karyotypic evolution of Channa species is associated with a decrease in the number of chromosomes and an increase in the number of chromosome arms, mainly due to fusions and pericentric inversions. The data obtained support the assumption that fragmentation and isolation of populations, especially of continental islands, contribute to the karyotypic diversification of snakeheads and are of interest for further cytogenetic studies of Channidae.

Cryptic species diversity and molecular diagnosis of Channa orientalis; an endemic freshwater fish of Sri Lanka

Fish genetic resources and diversity are very important aspects of environmental management and fisheries and are vital for making decisions on their commercial exploitation as well as conservation. The snakehead fishes in the world have significant economic importance as food and ornamental fish. A clear understanding of species’ taxonomic status and genetic diversity is important for the utilization and implementation of conservation and management practices. Channa orientalis is a snakehead endemic to Sri Lanka that is heavily utilized in the ornamental fish export trade. Its genetic diversity has not yet been fully understood and it is difficult to distinguish it from closely resembling species. Therefore, we examined the genetic diversity of C. orientalis and developed a DNA-based marker that permits accurate, low cost, and reliable identification of C. orientalis. Determination of genetic diversity was mainly carried out through genetic analysis of the mitochondrial cytochrome c oxidase subunit 1 (MT-CO1) gene. The development of the DNA-based marker for the identification of C. orientalis was done through Polymerase Chain Reaction and Restriction Fragment Length Polymorphism (PCR-RFLP) analysis. Our analyses confirmed the presence of two distinct genetically divergent and geographically separated lineages of C. orientalis in Sri Lanka. The fast cost-effective gel-based PCR-RFLP marker method developed by us was successful in diagnosing C. orientalis from its closely resembling species. Thus, we believe our findings on the cryptic diversity and diagnostic methods will have important implications for the conservation and management of this endemic species.

Aenigmachannidae, a new family of snakehead fishes (Teleostei: Channoidei) from subterranean waters of South India

Pronounced organism-wide morphological stasis in evolution has resulted in taxa with unusually high numbers of primitive characters. These ‘living fossils’ hold a prominent role for our understanding of the diversification of the group in question. Here we provide the first detailed osteological analysis of Aenigmachanna gollum based on high-resolution nano-CT scans and one cleared and stained specimen of this recently described snakehead fish from subterranean waters of Kerala in South India. In addition to a number of derived and unique features, Aenigmachanna has several characters that exhibit putatively primitive conditions not encountered in the family Channidae. Our morphological analysis provides evidence for the phylogenetic position of Aenigmachanna as the sister group to Channidae. Molecular analyses further emphasize the uniqueness of Aenigmachanna and indicate that it is a separate lineage of snakeheads, estimated to have split from its sister group at least 34 or 109 million years ago depending on the fossil calibration employed. This may indicate that Aenigmachanna is a Gondwanan lineage, which has survived break-up of the supercontinent, with India separating from Africa at around 120 mya. The surprising morphological disparity of Aenigmachanna from members of the Channidae lead us to erect a new family of snakehead fishes, Aenigmachannidae, sister group to Channidae, to accommodate these unique snakehead fishes.

Into Africa via docked India: a fossil climbing perch from the Oligocene of Tibet helps solve the anabantid biogeographical puzzle

The northward drift of the Indian Plate and its collision with Eurasia have profoundly impacted the evolutionary history of the terrestrial organisms, especially the ones along the Indian Ocean rim. Climbing perches (Anabantidae) are primary freshwater fishes showing a disjunct south Asian-African distribution, but with an elusive paleobiogeographic history due to the lack of fossil evidence. Here, based on an updated time-calibrated anabantiform phylogeny integrating a number of relevant fossils, the divergence between Asian and African climbing perches is estimated to have occurred in the middle Eocene (ca. 40 Ma, Ma: million years ago), a time when India had already joined with Eurasia. The key fossil lineage is †Eoanabas, the oldest anabantid known so far, from the upper Oligocene of the Tibetan Plateau. Ancestral range reconstructions suggest a Southeast Asian origin in the early Eocene (ca. 48 Ma) and subsequent dispersals to Tibet and then India for this group. Thereby we propose their westbound dispersal to Africa via the biotic bridge between India and Africa. If so, climbing perch precursors had probably followed the paleobiogeographical route of snakehead fishes, which have a slightly older divergence between African and Asian taxa. As such, our study echoes some recent molecular analyses in rejecting the previously held “Gondwana continental drift vicariance” or late Mesozoic dispersal scenarios for the climbing perches, but provides a unique biogeographical model to highlight the role of the pre-uplift Tibet and the docked India in shaping the disjunct distribution of some air-breathing freshwater fishes around the Indian Ocean.

Molecular identification and genetic diversity of Gnathostoma spinigerum larvae in freshwater fishes in southern Lao PDR, Cambodia, and Myanmar.

Gnathostomiasis, an emerging food-borne parasitic zoonosis in Asia, is mainly caused by Gnathostoma spinigerum (Nematoda: Gnathostomatidae). Consumption of raw meat or freshwater fishes in endemic areas is the major risk factor. Throughout Southeast Asia, including Thailand, Lao PDR, Cambodia, and Myanmar, freshwater fish are often consumed raw or undercooked. The risk of this practice for gnathostomiasis infection in Lao PDR, Cambodia, and Myanmar has never been evaluated. Here, we identified larvae of Gnathostoma species contaminating freshwater fishes sold at local markets in these three countries. Public health authorities should advise people living in, or travelling to, these areas to avoid eating raw or undercooked freshwater fishes. Identification of larvae was done using molecular methods: DNA was sequenced from Gnathostoma advanced third-stage larvae recovered from snakehead fishes (Channa striata) and freshwater swamp eels (Monopterus albus). Phylogenetic analysis of a portion of the mitochondrial cytochrome c oxidase subunit I gene showed that the G. spinigerum sequences recovered from southern Lao PDR, Cambodia, and Myanmar samples had high similarity to those of G. spinigerum from China. Sequences of the nuclear ribosomal DNA internal transcribed spacer 2 region closely resembled sequences of G. spinigerum from Thailand, Indonesia, the USA, and central Lao PDR. This is the first molecular evidence of G. spinigerum from freshwater fishes in southern Lao PDR, Cambodia, and Myanmar.

Cytochrome-C Oxidase I Gene-Based Genetic Divergence and Molecular Phylogeny Among the Species of Fish Genus Channa

The freshwater snakehead fishes are members of the family Channidae and order Perciform, which are native to parts of Africa and Asia with widespread distributions. The present study was aimed to assess the genetic variability and molecular phylogeny among 22 species of genus Channa, viz. C. argus, C. asiatica, C. aurantimaculata, C. bankanensis, C. barca, C. bleheri, C. burmanica, C. diplogramma, C. gachua, C. lucius, C. maculata, C. marulius, C. melasoma, C. micropeltis, C. orientalis, C. ornatipinnis, C. panaw, C. pleurophthlama, C. pulchra, C. punctata, C. stewartii and C. striata, using partial 513 bp long consensus sequence of cytochrome-c oxidase subunit I mitochondrial DNA retrieved from NCBI (March, 2018). The study showed 281 haplotypes with 267 variable sites (i.e., 33 singletons and 234 parsimoniously informative) in nucleotide positions. Most of the sequences were A + T rich, except in C. argus which is G + C rich (50.25%). The NJ and MP trees showed similar topology with discrete branches, each as a distinct species of the Channa with slight differences in bootstrap values. Maximum K2P distance was found between the C. ornatipinnis and the C. melasoma. The maximum numbers of polymorphic loci were present in C. gachua.

Barcoding snakeheads (Teleostei, Channidae) revisited: Discovering greater species diversity and resolving perpetuated taxonomic confusions.

Snakehead fishes of the family Channidae are predatory freshwater teleosts from Africa and Asia comprising 38 valid species. Snakeheads are important food fishes (aquaculture, live food trade) and have been introduced widely with several species becoming highly invasive. A channid barcode library was recently assembled by Serrao and co-workers to better detect and identify potential and established invasive snakehead species outside their native range. Comparing our own recent phylogenetic results of this taxonomically confusing group with those previously reported revealed several inconsistencies that prompted us to expand and improve on previous studies. By generating 343 novel snakehead coxI sequences and combining them with an additional 434 coxI sequences from GenBank we highlight several problems with previous efforts towards the assembly of a snakehead reference barcode library. We found that 16.3% of the channid coxI sequences deposited in GenBank are based on misidentifications. With the inclusion of our own data we were, however, able to solve these cases of perpetuated taxonomic confusion. Different species delimitation approaches we employed (BIN, GMYC, and PTP) were congruent in suggesting a potentially much higher species diversity within snakeheads than currently recognized. In total, 90 BINs were recovered and within a total of 15 currently recognized species multiple BINs were identified. This higher species diversity is mostly due to either the incorporation of undescribed, narrow range, endemics from the Eastern Himalaya biodiversity hotspot or the incorporation of several widespread species characterized by deep genetic splits between geographically well-defined lineages. In the latter case, over-lumping in the past has deflated the actual species numbers. Further integrative approaches are clearly needed for providing a better taxonomic understanding of snakehead diversity, new species descriptions and taxonomic revisions of the group.

Genomic Organization of Repetitive DNA Elements and Its Implications for the Chromosomal Evolution of Channid Fishes (Actinopterygii, Perciformes).

Channid fishes, commonly referred to as “snakeheads”, are currently very important in Asian fishery and aquaculture due to the substantial decline in natural populations because of overexploitation. A large degree of chromosomal variation has been found in this family, mainly through the use of conventional cytogenetic investigations. In this study, we analyzed the karyotype structure and the distribution of 7 repetitive DNA sequences in several Channa species from different Thailand river basins. The aim of this study was to investigate the chromosomal differentiation among species and populations to improve upon the knowledge of its biodiversity and evolutionary history. Rearrangements, such as pericentric inversions, fusions and polyploidization, appear to be important events during the karyotypic evolution of this genus, resulting in the chromosomal diversity observed among the distinct species and even among populations of the same species. In addition, such variability is also increased by the genomic dynamism of repetitive elements, particularly by the differential distribution and accumulation of rDNA sequences on chromosomes. This marked diversity is likely linked to the lifestyle of the snakehead fishes and their population fragmentation, as already identified for other fish species. The karyotypic features highlight the biodiversity of the channid fishes and justify a taxonomic revision of the genus Channa, as well as of the Channidae family as a whole, as some nominal species may actually constitute species complexes.

Calibrating snakehead diversity with DNA barcodes: expanding taxonomic coverage to enable identification of potential and established invasive species.

Detecting and documenting the occurrence of invasive species outside their native range requires tools to support their identification. This can be challenging for taxa with diverse life stages and/or problematic or unresolved morphological taxonomies. DNA barcoding provides a potent method for identifying invasive species, as it allows for species identification at all life stages, including fragmentary remains. It also provides an efficient interim taxonomic framework for quantifying cryptic genetic diversity by parsing barcode sequences into discontinuous haplogroup clusters (typical of reproductively isolated species) and labelling them with unique alphanumeric identifiers. Snakehead fishes are a diverse group of opportunistic predators endemic to Asia and Africa that may potentially pose significant threats as aquatic invasive species. At least three snakehead species (Channa argus, C. maculata, and C. marulius) are thought to have entered North America through the aquarium and live-food fish markets, and have established populations, yet their origins remain unclear. The objectives of this study were to assemble a library of DNA barcode sequences derived from expert identified reference specimens in order to determine the identity and aid invasion pathway analysis of the non-indigenous species found in North America using DNA barcodes. Sequences were obtained from 121 tissue samples representing 25 species and combined with public records from GenBank for a total of 36 putative species, which then partitioned into 49 discrete haplogroups. Multiple divergent clusters were observed within C. gachua, C. marulius, C. punctata and C. striata suggesting the potential presence of cryptic species diversity within these lineages. Our findings demonstrate that DNA barcoding is a valuable tool for species identification in challenging and under-studied taxonomic groups such as snakeheads, and provides a useful framework for inferring invasion pathway analysis.

Length-weight and length-length relationship of three species of snakehead fish, Channa diplogramma, C. marulius and C. striata from the riverine reaches of Lake Vembanad, Kerala, India

The length-weight relationship (LWR) and length-length relationships (LLR) of three snakehead fishes, Channa diplogramma, C. marulius and C. striata, exploited by small-scale fishers in the riverine reaches of Lake Vembanad, Kerala were studied using the allometric growth equation Y = aXb. Our analysis shows that the LWR of C. diplogramma and C. marulius is nonisometric with exponents much smaller than the cubic value (b = 3), while that of C. striata is isometric. Channa marulius showed a definite change in LWR with size, with smaller fish growing with positive allometric exponents (b greater than 3) and larger individuals having negative allometric relationship (b less than 3), indicating a possible age-related change in growth pattern. In the case of LLR, all three snakehead species showed non-isometric growth patterns. The caudal fin did not grow substantially with increasing fish length.

Relationships and biogeography of the fossil and living African snakehead fishes (Percomorpha, Channidae, Parachanna)

ABSTRACT The ability to determine relationships of Cenozoic fossil fishes relies heavily on having osteological information from their extant relatives with which to compare the fossil remains. For many higher teleost fishes, these osteological data do not exist. For example, †Parachanna fayumensis, from Eocene and Oligocene deposits of Egypt, was placed in the Recent snakehead genus Parachanna, but in the absence of data from extant members of the genus, this hypothesis could not be tested. Three other fossil channids were each given their own new genera, without an analysis of their relationships to one another or to the living genera. In order to properly test the relationships of these fossil species, the osteological information for living species is needed. This paper documents the osteology of the African snakehead, Parachanna obscura, for the first time. The documentation of the osteology forms the basis of a phylogenetic study of the relationships of the African fossil, †Parachanna fayumensis, with living African snakeheads. This study supports the generic placement of †P. fayumensis as being correct, as well as reciprocal monophyly of the two living snakehead genera and monophyly of the family. Monophyly of the Asian genus Channa and the relationships of the other fossil species need to be further assessed using a broader range of species, but this preliminary study is an essential first step. Apomorphic osteological characters are given for the family and genera, as well as a discussion of the biogeographical relationships of species of Parachanna.

A reappraisal of the evolution of Asian snakehead fishes (Pisces, Channidae) using molecular data from multiple genes and fossil calibration

Freshwater snakehead fishes (Channidae) provide an interesting target for phylogenetic analysis for the following reasons, their unusual biology, potential for cryptic diversity and availability of a good fossil record. Here, a multi-locus molecular phylogeny was constructed and calibrated using two fossil dates to estimate divergence times within the family. Sampling aimed to explore interspecific divergence of Channa species across Southeast Asia and intra-specific variation where species possessed natural geographical ranges that were extensive. Results contradict divergence times estimated previously independently from single locus mitochondrial data or the fossil record and suggest that after divergence from African taxa 40–50 Ma, evolution of Asian snakeheads has been heavily influenced by multiple broad scale dispersal events across India and Southeast Asia. A similar pattern of divergence within multiple clades suggests that west–east dispersal was limited for many taxa during the Miocene. Deep intra-specific divergence was inferred for C. striata, indicating that long historical periods of isolation (∼8 Ma) have not resulted in the evolution of reproductive isolation within this species. Results support suggestions that C. marulia like fishes in northern Cambodia may constitute an undescribed species, and that Indian C. diplogramma warrants taxonomic recognition as being distinct from Southeast Asian C. micropeltes, with the two taxa last sharing a common ancestor in the mid- to late-Miocene.

Migration history of air-breathing fishes reveals Neogene atmospheric circulation patterns

The migration history of an air-breathing fish group (Channidae; snakehead fishes) is used for reconstructing Neogene Eurasian precipitation and atmospheric circulation patterns. The study shows that snakeheads are sensitive indicators of summer precipitation maxima in subtropical and temperate regions, and are present regularly if the wettest month exceeds 150 mm precipitation and 20 8C mean temperature. The analysis of 515 fossil freshwater fish deposits of the past 50 m.y. from Africa and Eurasia shows two continental-scale migration events from the snakeheads’ center of origin in the south Himalayan region, events that can be related to changes in the Northern Hemisphere circulation pattern. The first migration, ca. 17.5 Ma, into western and central Eurasia may have been caused by a northward shift of the Intertropical Convergence Zone that brought western Eurasia under the influence of trade winds that produced a zonal and meridional precipitation gradient in Europe. During the second migration, between 8 and 4 Ma, into Africa and East Asia, snakeheads reached their present-day distribution. This migration could have been related to the intensification of the Asian monsoon that brought summer precipitation to their migratory pathways in East Africa‐Arabia and East Asia.