Age,growth and reproduction of a population of spined loach,Cobitis taenia (L.)

751 specimens of spined loach, Cobitis taenia (L.), were collected and studied over the period October 1972–October 1974. The majority of the males were found to die at the end of the 2+ season and the majority of the females at the end of the 3+ season. Back calculations for length at age were carried out from otolith measurements and found to correspond closely to actual measurements. Males and females grew similarly up until the end of their 1+ year of life, with most rapid growth occurring in the first year. Females continued to grow rapidly up to 2+, the rate then being reduced; the males showed an increasingly reduced rate of growth after the 1+ year. Ford Walford plots gave L∞ results which corresponded closely in most cases with the largest observed specimens. Gonad development and maturation of males and females followed different patterns. Males matured during the 1 + year at a length of approximately 40 mm from which point their dry gonad weight remained at a steady 0.02–0.03% of the body weight. Females appeared to mature a year later and their ovary weight and oocyte diameter increased with fish age. A seasonal cycle in gonad development was evident in females. There was a greater proportion of females to males, especially in the older age group. Spawning time was estimated to be between June and July.     

Some factors influencing the microdistribution of a population of spined loach,Cobitis taenia (L.)

Monthly observations (October 1972–October 1974) of a natural population of spined loach, Cobitis taenia (L.), in the River Great Ouse at Newport Pagnell indicated a patchy microdistribution, which varied seasonally. The distribution was clearly linked to the areas of fine substrate, which altered seasonally in position.Flow rates measured at monthly intervals in areas where Cobitis taenia (L.) were present were found to be approximately half those of areas in which Cobitis taenia L. were not found (0.148 m/sec and 0.293 m/sec). Substrate samples from areas where Cobitis taenia L. were present had a fine organic component, in contrast to the remainder of the river bed, which consisted of hard-packed gravel.In the laboratory, choice chamber experiments demonstrated a positive selection for fine organic deposits.     

Banded karyotype of spined loach Cobitis taenia and triploid Cobitis from Poland

The present work provides new data on the banding pattern of diploid Cobitis taenia and its triploid hybrid females, which belong to the diploid–polyploid complex in the Vistula River tributary. C-banding, silver-staining (Ag), and fluorescent staining with chromomycin A₃ techniques were used to describe the diploid and triploid karyotype. The karyotype of Cobitis taenia of 2n=48 was characterised by one pair of NOR-bearing subtelocentric chromosomes and at least four chromosomes with CMA₃-positive sites. The C-positive heterochromatin was present in the centromeres of almost all chromosomes and the pericentromeric regions of several metacentric and submetacentric chromosomes. The triploid females of 3n=74 had two pairs of chromosomes with active NORs. The NORs-sites were located terminally on two biarmed and two uniarmed chromosomes. The CMA₃-staining revealed at least six A₃-positive sites. The C-banded and A₃-stained triploid karyotype was composed of haploid set of Cobitis taenia and diploid set of unidentified species, so heterochromatin pattern confirmed the possibility of their hybrid origin. The characteristics of banded diploid and triploid karyotype, and the hypothetical karyotype of an unknown species of 2n=50 is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Infection of the spined loach, Cobitis taenia (L.), by the digenean, Allocreadium transversale (Rud.)

The digenean Allocreadium transversale (Rud.) has been recorded once previously in Great Britain. During work on the feeding of Cobitis taenia (L.) it was found to be a frequently occurring gut parasite. Great variation occurred in its seasonal level of incidence and mean worm burden, indicating a rapid turnover and replacement of the parasite in the fish. No significant relationship could be shown between the level of infection and age of the fish; but a highly significant relationship occurred between incidence of infection and fish age.     

Replication banding patterns in the spined loach,Cobitis taenia L. (Pisces,Cobitidae)

The chromosomal complement of Cobitis taenia was analysed by replication banding techniques to determine whether there were specific patterns that could allow distinction of the different chromosomes. The diploid chromosome number of 2n = 48 is diagnostic of this species. In vivo 5-bromodeoxyuridine (5-BrdU) incorporation induced highly reproducible replication bands. Most of the chromosome pairs were distinguishable on the base of their banding patterns. The karyotype, consisting of five pairs of metacentrics, nine pairs of submetacentrics and 10 pairs of subtelocentrics and acrocentrics, was confirmed. C-banding and replication banding patterns were compared, and heterochromatin was both early and later replicating. C-positive heterochromatin in centromeric regions was mainly early replicating, but that located in pericentromeric regions was late replicating. Most of the late-replicating regions found interstitially were C-band negative. The results obtained so far for combined chromosomal staining methods of C. taenia and other Cobitis fish species are discussed.     

Comparative histology of the testes of the spined loach Cobitis taenia L. and natural allotetraploids of Cobitis (Pisces, Cobitidae)

The spined loach Cobitis taenia L. creates exclusively diploid and mixed diploid–polyploid populations. Allotriploid females, which co-exist with C. taenia or C. elongatoides and a few tetraploid males and females dominate in most Cobitis mixed populations. They reproduce gynogenetically and produce triploid eggs that are stimulated to development by sperm from Cobitis males. Some of these eggs are fertilized, which leads to the production of bisexual tetraploids. Males of C. taenia (2n = 48) from a diploid population in Lake Klawój, Northern Poland (46 individuals) and from a mixed Cobitis population in the Bug River, Eastern Poland (7 individuals), and three tetraploid males (4n = 98) from the same mixed population were examined. All the fish were analyzed karyologically and histologically. Tubules with cysts of the testes of C. taenia from both populations were filled with germ cells at various developmental stages. Among fishes from Lake Klawój sperm maturation in batches simultaneous with the batch spawning of C. taenia females was found. The testes of the loach C. taenia, from a mixed population in the Bug River, were filled with spermatozoa over the entire reproductive season. Sperm maturation in batches was not observed. Sperm maturation in batches seems to be only connected with a few diploid males in this population. So, a continuous process of spermatogenesis in their testes is required. Only in the testes of all tetraploid Cobitis males were cells characteristic of the early stages of spermatogenesis observed, i.e. without spermatids and spermatozoa. Furthermore, the histological sections of the testis of a male captured in August, revealed fragments with connective tissue between the germ cells. However the participation of tetraploid, infertile Cobitis males in the process of reproduction in the investigated mixed population remains controversial. The results obtained so far, reveal that even the infertile sperm of tetraploid males may induce gynogenesis in Cobitis triploid females.     

Biochemical genetic evidence for a hybrid origin of spined loach, Cobitis taenia taenia, in Japan

Starch gel electrophoresis and isoelectric focusing were performed on sarcoplasmic proteins from three taxa of cobitid fish in Japan, Cobitis biwae , C. taenia striata and C. l. taenia . These taxa are hardly distinguishable from each other by external appearance or morphological characters. Electrophoretic patterns of sarcoplasmic proteins from white muscle suggested fixed allelic differences between C. biwae and C. t. striata at two protein-coding loci (parvalbumin and creatine kinase). The third taxon, C. t. taenia , showed composite banding patterns with respect to the above two proteins, suggesting C. t, taenia originated from hybridization between C. biwae and C, t. striata . Previously, C. t, taenia and C. t. striata were believed to have shared a common ancestor subsequent to their divergence from C. biwae .     

Reproduction of spined loach, Cobitis taenia, (Cypriniformes; Cobitidae) under laboratory conditions

A simple and reliable method is described for breeding spined loach, Cobitis taenia Linnaeus, 1758, under laboratory conditions. Three 40 L aquaria were each provided with a thick tuft of moss placed on top of a gauze-covered plastic box to serve as a spawning site. Ovoposition always occurred in the most densely available vegetation. The nonadhesive eggs fell through the gauze into the box. The spined loach is a fractional spawner releasing eggs in 14–18 batches at intervals of 2–23 (median 6) days. The total number of eggs per female was 2905–4282 during the reproductive season of 101–120 days. Yolks measured 1.14 ± 0.07 mm and eggs were 2.54 ± 0.22 mm in diameter. Total larval length was 5.03 ± 0.34 mm at hatching and 6.77 ± 0.34 mm at the beginning of exogenous feeding. The method allowed the rearing of numerous young for conservation measures and experimental investigations.     

A comparison of ventilation rates and haemoglobin concentrations between two species of loach,Noemacheilus barbatulus (L.) and Cobitis taenia (L.)

The mean haemoglobin concentration of Noemacheilus barbatulus was found to be significantly higher than that of Cobitis taenia. Under conditions of reduced oxygen saturation, both species achieved maximum rates of ventilation at 10% 0₂. An alteration in opercular beat pattern accounted for the increased ventilation rates between 20% and 30%. A slightly higher tolerance for lowered oxygen concentrations was demonstrated by Cobitis taenia with disorientation and aerial respiration occurring at lower oxygen tensions than in Noemacheilus barbatulus.     

Spatial and ontogenetic variability in the microhabitat use of stream-dwelling spined loach (Cobitis taenia) and stone loach (Barbatula barbatula)

Ontogenetic and spatial variability in microhabitat use of spined loach Cobitis taenia (Linnaeus), considered as one species for the purposes of this study, and stone loach Barbatula barbatula (Linnaeus) were examined in the River Great Ouse basin, England, using multivariate and habitat suitability methods, including a technique for handling spatial variation in collections of preference curves. Distinct ordinations of spined age classes and stone loach developmental stages, respectively, in canonical correspondence analysis of species × variables × samples relationships suggest that the two species occupy completely different microhabitats; however, young‐of‐the‐year spined loach occurred more often than expected with all developmental stages of stone loach except young larvae. Water velocity and filamentous algae were the most influential microhabitat variables, the latter decreasing in importance with increasing age of both fish species. Preferred water velocities generally decreased with age in spined loach and increased in stone loach, with substratum size generally increasing with fish age in both species. Spatial variation in microhabitat preferences was great in both species but less so in the spined loach, suggesting that limited plasticity in habitat use could account, at least in part, for the latter species’ limited distribution and abundance in the catchment. Preference curves for a species, if generated and verified for all life intervals and all seasons, could be used as a management tool for a given stream or sector of river basin. But preference curves should be generated for each location to ensure that river management decisions with regard habitat and species conservation consider local‐level species requirements. Thus, a multi‐(eco)species and multi‐scale approach is required in habitat suitability assessments.     

An analysis of a specialized feeding mechanism of the spined loach, Cobitis taenia (L.), and a description of the related structures

Aquarium observations and gut analyses of the spined loach, Cobitis taenia , suggested the use of a technique by which small items of food could be removed from an almost constant stream of substrate, taken in through the mouth and passed out through the gills. The small amount of indigestible substrate present in the gut contents indicated a very high efficiency level for the process. Photographic investigation revealed a two phased feeding cycle based on the respiratory pumps, consisting of an induction/separation phase and an expulsion/swallowing phase. A morphological study showed large concentrations of mucus secreting goblet cells on the roof of the anterior buccopharyngeal cavity. Other morphological adaptations were noted which resulted in a narrowing of the anterior buccal cavity and a widening of the posterior part along with an increase in the distance between the gill rakers. In the light of this information a method of separation based on differential densities of particles being trapped in a constant stream of mucus is postulated.     

European colonization by the spined loach (Cobitis taenia) from Ponto-Caspian refugia based on mitochondrial DNA variation

In the last 20 years, new species, asexual reproduction, polyploidy and hybridization have all been reported within the genus Cobitis. An understanding of the current distribution and baseline phylogeographical history of 'true' nonhybrid Cobitis species is crucial in order to unravel these discoveries. In the present work, we investigated the phylogeography of the spined loach, Cobitis taenia, using 1126 bp of the mitochondrial cytochrome b gene from 174 individuals collected at 47 sites. In total, 51 haplotypes that differed at 49 positions (4.35%) were detected. We deduce that C. taenia survived European glaciations in at least three refugees in the Ponto-Caspian area. Two of these refugees each provided a major lineage that recolonized Europe in separate directions: one westward to England and the other spreading north into Russia before moving west. A third (minor) lineage that contributed little to the recolonization of Europe was also revealed--remaining near its Black Sea refuge. However, more recent history was difficult to resolve with colonization from a more western refugium during the last glacial maximum (LGM) a distinct possibility. Nested clade analysis indicates a pattern of restricted gene flow with isolation by distance at the first two levels and overall. Unlike many other European freshwater fish species, the Danube is not part of the current distribution of C. taenia, nor was it used as either a refuge or a source of colonization of Europe. Low genetic diversity within C. taenia suggests that its colonization of Europe is relatively recent. Demographic analyses revealed a history of recent expansion and isolation by distance.     

Genetic structure of a diploid-polyploid complex of the spined loach Cobitis taenia (Cypriniformes: Cobitidae) from the middle Dnieper bassin

Biochemical genetic typing and cytometry showed that polyploid females account for 87% of the spined loach Cobitis taenia population from the middle Dnieper basin. The polyploidy series included triploids, tetraploids, and, possibly, a few pentaploids. A characteristic feature of the genetic structure of polyploids was that their genetic variation was due to the clonal variation in the haploid portion of the genome originating from Cobitis sp. and to polymorphism of the diploid portion originating from C. taenia. The results are discussed with regard to comparative evolution of alloploid complexes in fish and terrestrial vertebrates.     

Reproductive and habitat isolation between two populations of the striated spined loach

Synopsis This study deals with ecological differences influencing reproductive isolation between two sympatric populations, the small and middle forms, of the striated spined loach. As natural hybrids are rare, isolation between these populations must be complete. There are some differences in their reproduction and early life history. Both reproduce in temporary waters flooded in summer, but their spawning sites are isolated from each other. The small form spawns in shallow areas of still water around irrigation creeks or fens. On the other hand, the middle form spawns in slow-flowing ditches. Spawning site segregation between them is an important isolating mechanism, and is a by-product of adaptation in their early life history.     

Diversity of European spined loaches (genus Cobitis L.): an update of the geographic distribution of the Cobitis taenia hybrid complex with a description of new molecular tools for species and hybrid determination

Although the unique features of asexual reproduction and hybridization among European spined loaches (genus Cobitis ) have recently attracted the attention of conservation biologists, faunists and evolutionary biologists, the research has suffered from uncertain identification of specimens and their genomes because of the extreme morphological similarity of all the species within the hybrid complex. In this article, a Europe-wide study is reported, which was performed on samples collected by several research teams. Several complementary methodologies, such as allozyme analysis, karyotyping, flow cytometry and DNA sequencing allowed us to confirm or reject the existence of all previously reported species and their hybrids as well as to uncover several new hybrid biotypes. The biogeography of all the known biotypes, that is, parental species and hybrid biotypes, has been summarized here and the taxonomic position of two undescribed putative species mentioned in previous publications has been established. New polymerase chain reaction restriction fragment length polymorphism markers for species determination have further been developed and applied, which would allow the unambiguous identification of parental species and their genomes in the known hybrid biotypes within the complex.     

Development of nine polymorphic microsatellite loci in the spined loach, Cobitis taenia, and cross-species amplification in the related species C. elongatoides, C. taurica and C. tanaitica

Nine polymorphic microsatellite loci were developed for the spined loach, Cobitis taenia (Teleostei: Cobitidae). The loci were validated using 50 individuals from a population in Belgium. Moderate to high levels of polymorphism were detected (two to 11 alleles). In addition, most markers amplified successfully in three closely related taxa that are known to hybridize with C. taenia: C. elongatoides, C. taurica and C. tanaitica. Some of the loci are most likely diagnostic among species. These markers will be valuable for the study of the historical and contemporary interactions within C. taenia and the Cobitis species complex.     

Genetic relationships among the Japanese and Korean striated spined loach complex (Cobitidae: Cobitis) and their phylogenetic positions

The striata complex, a group of spined loaches included in the genus Cobitis and characterized by a striped coloration pattern on the lateral midline, is distributed in rivers in northeastern Asia to western Japan. The complex comprises 2 continental species (Cobitis tetralineata and Cobitis lutheri) and 3 Japanese races of species rank (large race, middle race, and small race), the small race further comprising 6 local forms of subspecific rank (Tokai form, Biwa form, Yodo form, Sanyo form, San-in form, and Kyushu form). Previous karyological studies have revealed that the large race is an allotetraploid, the others being diploid. In this study, mitochondrial (mt) DNA analyses were conducted for 30 diploid populations of the Cobitis striata complex from Japan and Korea to examine: (1) their phylogenetic relationships and the position of the complex among the major lineages of Cobitis; and (2) the genetic relationships among the Japanese and Korean populations. The results, based on cytochrome b sequences (724 base pairs) analyzed with those of the main lineage of European and Japanese Cobitis, indicated that the striata complex should be considered as a monophyletic group, which evolved in northeastern Asia. Initially considered as a subspecies of Cobitis taenia, widely distributed from Europe to Asia, the striata complex does not have a sister-relationship with the former. Although the Korean species C. tetralineata was formerly believed to be closely related to the middle race in Japan, and a second continental species, C. lutheri, closely related to the Kyushu or San-in forms of the small race in Japan, the trees resulting from the present study revealed that the two Korean species were clustered with each other and separated from all Japanese races.