Study on some biological and feeding factors of spined loach Cobitis taenia from the Babolrud, Talar and Siahrud Rivers

Spined loach, Cobitis taenia, is a predominant fish in the river systems of the southern Caspian Sea basin. Although there is evidence of the geographical divergence of this taxon, but no information is available on morphological differences within the species populations. This study was designed to evaluate some biological factors including; morphometric and meristic characters, length-weight, age-growth, condition factor, diet, reproduction, variation and differentiation, in the Babolrud, Talar and Siahrud Rivers in south of the Caspian Sea basin. Age, sex ratio, fecundity, ova diameter and gonadosomatic index were estimated. Also, regression analyses was tested the relation between fecundity and fish length, weight, gonad weight, and also age. Totally 858 fish of which 721 were matures, were collected from these rivers by electrofishing. 37 morphometric characters, 9 meristic characters and 78 truss network system characters were estimated. Resulats of DFA analysis based on data of morphometric and meristic showd that these populations are highly (94.5%) varios from each other. In discriminate function analysis, the proportion of individuals correctly classified into their original groups was 61%, 65.4% and 86.5% for upstream and downstream, respectively. Clustering based on Euclidean distances among groups of centroids using an UPGMA and also principal component analysis’ results for morphometric data indicated that these populations from these three rivers were clearly distinct from each other. Regression equations between length and weight in these three populations were significantly different from Folton factor (b=3), that showed the fish has a negative Alometric growth process. Condition factor was estimated between 0.8912 to 1.2736 and 0.8131 to 1.4489 for males and females, respectively. Sex ratio (female: male) in these populations was 1.2816:1. The difference between the number of females and males was significant and females were more than males. The female and male specimens reach maturity by Tl more than 40 and 30 mm and at the age of 2+ and 1+, respectively. The mean of ova diameter was 0.5824±0.2882. The spawning took place from May to late July, at the water temperature from 18.7 to 24.0°C. The GSI values average at the beginning of the reproduction period was about 9%, with ranged from 2 to 26% in ripe mature females. The absolute and relative fecundity were 2109±792 and 579±208 respectively. The absolute fecundity was significantly related to body weight and gonads weight. Based on the pattern of gonado-somatic index, it was concluded that this fish has prolong active reproductive period, which is a type of adaptation by short-lived small fishes to environmental conditions. The macroscopic and histological results showed that the female and male have 5 and 4 stages in their maturation process, respectively. The RLG index was about 0.4732, which showed the fish is a carnivorous species. Significant difference was observed between fishes with different length and diet. The main foods of the fish were Trichoptera, Chironomidae larvae and Ephemeroptera which were their prefered food as well, however it was estimated that the food selection and diet are affected by environmental conditions.

Balitora nantingensis (Teleostei : Balitoridae), a new hillstream loach from Salween drainage in Yunnan, Southwestern China

A new species of hillstream loach genus Balitora Gray is discovered from Nanting River drainage, which drains to Salween, Yongde County, Yunnan, China. Balitora nantingensis can be distinguished from B. lancangjiangensis by a triangle-like blotch in nape absent vs. present; area from halfway of pelvicfin ray to anus covered by scales vs. no scales in this area; the first pectoral ray thickened vs. not; posterior end of pelvic-fin surpasses middle point from posterior end of pelvic-fin base to anal-fin origin vs. the end from posterior extremity of pelvic-fins to anus; fewer lateral line scales, 59-64 vs. 66-70; slightly longer head, SL 4.2-5.1 (4.6) times of HL vs. 5.0-5.9 (5.4); slightly larger eyes, HL 4.0-6.4 (5. 1) times of eye diameter vs. 5.5-7.5 (6.3). Balitora nantingensis can be distinguished from B. burmanica by body rounded vs. slender in dorsal view; pointed snout vs. blunt in dorsal view; a deeper body, body depth 12.5-16.9% of SL vs. 10-12%; a deeper caudal-peduncle, length of caudal-peduncle 2.0-2.8 times of its depth vs. 3.0-4.2; a shorter caudal-peduncle, 14.3-15.6% of SL vs. 16-21%; the average ratio of body width at pelvic-fin origin and distance between inner pelvic-fin base 2.0 vs. 3.4; body width 1.1-1.5 times of body depth vs. 1.5-1.6; posterior end of pelvic-fin almost one or slight more than one eye diameter from anus vs. closet to anus; outside rays of pelvic-fin extend more posteriorly than inside rays and make posterior outline of fin acclivitous and straight vs. the outside ones almost equal to the inside ones and make the fin slightly convex.

Schistura cryptofasciata, a new loach (Cypriniformes : Balitoridae) from Salween drainage in Yunnan, Southwestern China

Schistura cryptofasciata is distinguished from its congeners by following combination characters: lateral line complete; 8(1)/(2) branched dorsal-fin rays; 9+8 branched caudal-fin rays; no sexual dimorphism; 8-11 indistinct bars, wider than interspaces; caudal-peduncle length 44.1-16.5% standard length, 68.1-90.4% dorsal head length; anterior portion of body scaleless; air bladder entirely embedded in bony capsule; lateral head length 104-122% dorsal head length; lower jaw with a notch at symphysis; lower lip completely interrupted; interorbital width 1.9-2.2 times eye diameter; pelvic-fin origin ventral to dorsal-fin origin; anus 1.0-1.5 eye diameters in front of anal-fin origin; the widest part of black basal caudal bar is the midlateral, and then the upper and lower ends.

Triplophysa rosa sp nov.: a new blind loach from China

A new blind loach of Triplophysa Rendahl 1933 was collected from a subterranean pool in a cave at Wulong County, Chongqing City, China, in November 2002. The new species, named Triplophysa rosa sp. nov., can be distinguished from its congeners by the following unique characters: eyes vestigial; 9 branched dorsal-fin rays; 12 branched pectoral-fin rays; 7 branched pelvic-fin rays; 6 branched anal-fin rays; 7 + 7 branched caudal-fin rays; distal margin of dorsal-fin concave; tip of pelvic-fin surpasses vertical level of anus; caudal-fin deeply forked; whole body scaleless and colourless. (c) 2005 The Fisheries Society of the British Isles.

Paracobitis nanpanjiangensis, a new loach (Balitoridae: Nemacheilinae) from Yunnan, China

A new species of the genus Paracobitis, Paracobitis nanpanjiangensis is described from tributaries of the Nanpanjiang River drainage in China. It is distinguished from its congeners, except P. oligolepis and P. wujiangensis, by body scaleless or with rudimentary scales (caudal peduncle with several deeply embedded scales). It can be differentiated from P. wujiangensis by the complete lateral line (vs. incomplete), lower dorsal crest reaching the vertical of origin of anal fin (vs. shorter and higher dorsal crest not reaching the base of anal fin). It is distinguished from P. oligolepis by the following characters: branched dorsal fin with 81/2 (a few 91/2) rays (vs. 91/2), interspaces between bars in front of dorsal fin conspicuously thinner than those behind (vs. vermiform markings), dorsal head without vermiform markings or obscure (vs. clearly vermiform markings on dorsal head), vertebrae 4 + 36-38 (vs. 4 + 39-41).

Detection of hybridization between two loach species (Paramisgurnus dabryanus and Misgurnus anguillicaudatus) in wild populations

Artificial interspecific hybrids between large scale loach P. dabryanus and tetraploid pond loach M. anguillicaudatus (Cobitidae, Cypriniformes) are viable. To detect the occurrence of possible natural hybridization, genetic analyses by using microsatellite markers were performed for natural populations of large scale loach and pond loach, the reciprocal laboratory hybrids, and "supposed hybrids" with ambiguous morphology. The fertility of the artificial hybrids was also tested. At one diagnostic microsatellite (Mac50), one out of 20 "supposed hybrids" was identified to be F-1 hybrid between the two loach species because it had the same genotype as that of the laboratory hybrids. The triploid hybrids between the two species were confirmed to be female-sterile. The results show that rare hybridization has occurred between diploid large scale loach and tetraploid pond loach in nature although it may have little effect in genetic introgression. This study is helpful for fish conservation and encourages further investigation on natural hybridization and introgression of loaches.

Triplophysa waisihani, a new species of nemacheiline loach from Northwest China (Pisces: Balitoridae)

Triplophysa waisihani, a new species of nemacheiline loach, is described from the Kax River, a tributary of the Ili River drainage in Xinjiang-Uighur Autonomous Region of Northwest China. It can be assigned to the T. labiata species group characterized by having widely separated anterior and posterior nostrils and no breeding tubercles on the sides of the head. Triplophysa waisihani resembles T. labiata and T. herzensteini, but differs from both in cephalic sensory-canal pattern and the structure of the gas bladder; from T. labiata in the structure of the pelvic girdle and absence or presence of the fourth basibranchial; and from T. herzensteini in the shape of the anal fin.

Gynogenesis and sex determination in large-scale loach Paramisgurnus dabryanus (Sauvage)

Gynogenesis was induced using heterologous sperms in large-scale loach, Paramisgurnus dabryanus (Sauvage), in which a ZW/ZZ sex determination was previously proposed. Three microsatellite loci were used to monitor exclusive maternal inheritance of gynogenetic progenies. The results showed that high percentages of meiogynogens were produced at 4 min post-fertilization and mitogynogens were produced at 18 min post-fertilization by heat shocks, while meiotic gynogenesis was induced by cold shocks within a wide period and high heterozygosity was even observed in gynogens produced at 24 min post-fertilization. The sex ratios of the F, progenies in three gynogenetic families were significantly deviated from 1: 1 expectation with a female bias in two families and a male bias in one family (P < 0.05), and the other four gynogenetic families showed approximate 1:1 sex ratios. Moreover, the self-mating between gynogenetic F, progenies and mating between gynogenetic F, progenies and normal individuals produced all-female progenies or identical proportions of females and males. The data of sex ratios generally confirmed that the sex determination in large-scale loach was determined by the putative ZW/ZZ system, and the possible reasons causing the biased sex ratios are discussed.

Triplophysa lixianensis, a new nemacheiline loach species (Pisces : Balitoridae) from the upper Yangtze River drainage in Sichuan Province, South China

Triplophysa lixianensis, a new nemacheiline loach species, is described from the Min Jiang of the upper Yangtze River drainage in Sichuan Province, South China. It can be separated from all other species of Triplophysa by having a unique combination of the following characters: posterior chamber of gas bladder greatly reduced or absent; caudal peduncle columnar with a roughly round cross- section at its beginning; anterior edge of lower jaw completely exposed or uncovered by lower lip; intestine short, forming a zigzag loop below stomach; dorsal- fin origin closer to caudal- fin base than to snout tip; pelvic fin inserted anterior to dorsal- fin origin; snout length 50.6 - 57.5 % of head length; eye diameter 12.3 15.4 % of head length; caudal peduncle length 25.1 - 27.1 % of standard length; anal fin with five branched rays; lower lip greatly furrowed with two thick lateral lobes; and body smooth or scaleless.

Effects of cell-cycle arrest agents on cleavage and development of loach (Misgurnus anguillicaudatus) embryos

The aim of this study was to determine the lowest concentration of nocodazole and colchicine to arrest blastomere division during the cleavage stage of loach embryos and to assess the reversibility and toxicity of the treatments in the treated embryos. Eight-cell loach embryos were incubated for 4, 8, 12, or 16 h in 1/10x Holtfreter supplemented with either nocodazole, an inhibitor of tubulin polymerization, or colchicine, an inhibitor of tubulin assembly. Complete arrest of cell cycle was observed, at a colchicine concentration of 0.996 mM and at a nocodazole concentration of 0.275 muM, respectively (the lowest effective concentration). No major morphological alteration in chromatin was observed. Reversibility and toxicity of both agents were dose and exposure period dependent. For both agents, prolonging cleavage arrest for more than 4 h (at the effective concentrations) is detrimental to development of embryos. Nocodazole treatment was less cytotoxic, whereas the concentrations of colchicine which induce cleavage arrest were detrimental to development beyond the blastula stage. Toxic effects beyond the blastula stage could be minimized for both agents by reducing the period of treatment and concentration.

The toxic effects of microcystin-LR on embryo-larval and juvenile development of loach, Misguruns mizolepis Gunthe

Microcystin-LR, a specific and potent hepatotoxin, was tested for its effects oil loach embryo-larval and juvenile development, The results of this study showed that loach embryos were more sensitive when exposed to microcystin-LR at a later than at an earlier stage of development, Juveniles were far less sensitive to MC-LR than were embryos and larvae. Mortality and developmental abnormality were proven to be dose-dependent and to be stage-specific sensitive. Among the abnormal changes noted were: pericardial edema and tubular heart, bradycardia, homeostasis, poor yolk resumption. small head, curved body and tail, and abnormal hatching, Liver and heart were the main targets of microcystin-LR toxicity. Ultrastructural analysis documented a complex set of sublethal effects of microcystin-LR on loach hepatocytes, chiefly including morphological alteration in nuclear and RER of loach liver cells. fit addition, microcystin-LR was lethal to loach juvenile in the subacute (7 days) exposure (LC50) = 593.3 mug/l). (C) 2002 Elsevier Science Ltd. All rights reserved.

The effect of ions and cryoprotectants upon sperm motility and fertilization success in the loach Misgurnus anguillicaudatus

The solutions commonly used to dilute or cryopreserve sperm are commonly composed of salts, buffers and cryoprotectants, which may affect gametes and subsequent fertilization success. Here, we have evaluated the effects of several cryoprotectants (methanol; MeOH, dimethyl sulfoxide; DMSO and dimethyl acetamide; DMA at concentrations of 0.25, 0.5 and 1%) and different ions (potassium, calcium and magnesium at concentrations of 1.25, 2.5, 5.0 and 10 mM) as sperm diluents upon sperm motility and fertilization success in the loach Misgurnus anguillicaudatus sperm. Our results demonstrated that DMSO (at 1%) decreased sperm motility while calcium and magnesium ions (from 2.5 mM) induced sperm aggregation and reduced sperm motility. Reduced fertilization rates were observed with potassium (from 1.25 mM), calcium (at 10 mM), magnesium (at 10 mM), DMA (at 1%), and DMSO (at 1%). We conclude that specific ions and cryoprotectants, and their relative concentrations caused effect upon loach gametes. These data are important to consider for the preparation of sperm diluents and activating solutions in order to manage gamete quality for artificial propagation. (C) 2012 Elsevier B.V. All rights reserved.

Vitrification of primordial germ cells using whole embryos for gene-banking in loach, Misgurnus anguillicaudatus

In gene-banking, primordial germ cells (PGCs), which are embryonic precursor cells of germ cells, are useful for cryopreservation because PGCs have a potential to differentiate into both eggs and sperm via germ-line chimera. Here, we have established vitrification methods for PGCs cryopreservation using 12- to 17-somite stage embryos in loach, Misgurnus anguillicaudatus, which were dechorionated, removed their yolk and injected with green fluorescent protein (GFP) -nos1 3'UTR mRNA to visualize their PGCs. In order to optimize cryopreservation medium for vitrification, the toxicity of cryoprotectants was analyzed. Different concentrations (2, 3, 4, 5 m) of dimethyl sulfoxide (DMSO), methanol (MeOH), ethylene glycol (EG) and propylene glycol (PG) as cryoprotectants were tested. Then, 5 m DMSO showed significantly-high toxicity. Based on this information, combinations called DMP (2 m (14.2% [v/v]) DMSO, 2 m (8.1% [v/v]) MeOH and 2 m (14.4% [v/v]) PG), DP (2 m (14.2% [v/v]) DMSO and 4 m (28.7% [v/v]) PG) and DE (2.1 m (15% [v/v]) DMSO and 2.7 m (15% [v/v]) EG) were evaluated for their toxicities and efficacy of PGCs cryopreservation using two types of equilibration step: direct immersion of cryopreservation media (one-step) and serial exposure to half and full concentration of cryopreservation media (two-step). Viable PGCs were obtained from post-thaw embryos which were cryopreserved by DP and DE with both 1- and 2-step equilibrations. Despite DP showing the highest toxicity, it gave the highest survival rate of embryonic cells after cryopreservation. When PGCs recovered from vitrified embryos were transplanted into host embryos at the blastula stage, the transplanted PGCs were able to migrate to a host genital ridge similarly as endogenous PGCs. It suggests that our methods could be useful to create a germ-line chimera for the production of gametes from PGCs of cryopreserved embryos.

Ecological aspects of New Forest streams, draining one of Britain's unique areas

This article describes the streams of this unique area of Britain and reviews the published and some unpublished information that is currently available. None of the rivers in the New Forest are more than 30 km long. Many reaches have been artificially straightened, channelized and regraded since the 1840's. The stream waters are typically base-poor, with low nutrient concentrations. Primary productivity and standing crops of algae are predictably low when compared with other streams carrying higher concentrations of minerals and nutrients. The earliest records on the macroinvertebrate fauna go back to the late 19th Century. By 1940, over 20 species of Trichoptera and 10 species of Plecoptera had been recorded, but only four species of Ephemeroptera. Twenty species of fish occur in the streams of the New Forest of which the most common are brown trout, minnow, bullhead, stone loach, brook lamprey and eel.