Purification, characterization and cDNA cloning of a trypsin from the hepatopancreas of snakehead (Channa argus)

A trypsin was purified from the hepatopancreas of snakehead (Channa argus) by ammonium sulfate fractionation and a series of column chromatographies including DEAE-Sepharose, Sephacryl S-200 HR and Hi-Trap Capto-Q. The molecular mass of the purified trypsin was about 22 kDa, as estimated by SDS-PAGE. The optimum pH and temperature of the purified trypsin were 9.0 and 40 °C, respectively. The trypsin was stable in the pH range of 7.5-9.5 and below 45 °C. The enzymatic activity was strongly inhibited by serine proteinase inhibitors, such as MBTI, Pefabloc SC, PMSF, LBTI and benzamidine. Peptide mass fingerprinting (PMF) of the purified protein obtained 2 peptide fragments with 25 amino acid residues and were 100% identical to the trypsinogen from pufferfish (Takifugu rubripes). The activation energy (Ea) of this enzyme was 24.65 kJ·M^− 1. Apparent K_m was 1.02 μM and k_cat was 148 S^− 1 for fluorogenic substrate Boc-Phe-Ser-Arg-MCA. A trypsinogen gene encoding 247 amino acid residues was further cloned on the basis of the sequence obtained from PMF and the conserved site peptide of trypsinogen together with 5′-RACE and 3′-RACE. The deduced amino acid sequence contains a signal peptide of 15 residues and an activation peptide of 9 amino acid residues with a mature protein of 223 residues. The catalytic triad His-64, Asp-107, Ser-201 and 12 Cys residues which may form 6 disulfide bonds were conserved. Compared with the PMF data, only 2 amino acid residues difference were identified, suggesting the cloned trypsinogen is quite possibly the precursor of the purified trypsin.     

The complete mitogenome of the snakehead Channa argus (Perciformes: Channoidei): genome characterization and phylogenetic implications

To better understand the phylogenetic status of the snakehead, Channa argus, we determined its complete mitogenome sequence using long-polymerase chain reaction and the direct sequencing method. The complete mitogenome sequence was 16,559?bp in length and contained 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, and 1 control region (D-loop), the gene composition/order of which was identical to that observed in most other vertebrates. This was the first report of the mitogenome sequence in suborder Channoidei. Phylogenetic relationships of 14 perciform suborders based on mitogenome sequences were reconstructed using Bayesian inference and maximum likelihood methods. The results strongly supported the monophyly of Perciformes and the snakehead, as a representative species of suborder Channoidei, formed the most basal branch having sister relationship with the clade containing all other analyzed perciform fishes. The further phylogenetic analyses of six channid species, based on cytochrome b gene, suggested that two channid genera constituted reciprocally monophyletic clades. In addition, the relaxed molecular clock method was used to estimate divergence dates among major suborders of Perciformes and major species in Channoidei.     

Ecotoxicological assessment of several estuarine zones in the south of Primorsky Krai

A complex ecotoxicological assessment of several estuarine zones of the Razdol’naya, Barabashevka, Ryazanovka, and Gladkaya rivers in Amur Bay, and the Artyomovka and Sukhodol rivers in Ussyry Bay was carried out in 2007 and 2008. Bottom sediments in the estuary of the Kievka river were collected in 2009. The content of petroleum hydrocarbons was determined in the water and bottom sediments of riverine and marine sectors of estuaries and the total toxicity of bottom sediments was tested with the use of mysids. Bottom sediments from a small river on Reineke Island, in which the concentrations of primary pollutants corresponded to the background values for the entire Peter the Great Bay, were used as a control. The content of organochlorine pesticides in the muscles and liver of the Redeye Eastern Redfin (Tribolodon brandtii) was determined and molecular biomarkers of oxidative stress in organs of the Redlip Mullet (Liza haematocheila) analyzed. The amount of wastewater and pollutants flowing into Amur Bay and Ussyry Bay with some of the rivers was estimated. The correlation coefficient between the toxicity of bottom sediments and the petroleum hydrocarbons content in them was found. The higher sediment toxicity in the riverine zone, as compared to the marine zone, proves the concept of the “river-sea” biogeochemical barrier.     

Mitochondrial DNA diversity in wild boar from the Primorsky Krai Region (East Russia)

We have studied the cytochrome B gene and control region DNA variability in 14 wild boars from the Primorsky Region, in the far east corner of Russia. Variability was low (π = 0.00248 overall) compared with the usual estimates in these loci, indicating that this is a rather closed population. Seven haplotypes were found, and one was identical to a Chinese wild boar. Phylogeographically, the sequences clustered among several Asian clades, primarily Chinese domestic pigs and Japanese and Chinese wild boars, and are positioned within the D2 clade reported by Larson et al. [ Science 307 , 2005; 1618 ]. Although North Korean pigs should be studied, our data suggest that the Primorsky mtDNA signature is absent from domestic pigs. Sequences are available through GenBank identifiers HM010461 – HM010488 .     

Haematological and serum biochemical characterization and comparison of wild and cultured northern snakehead (Channa argus Cantor, 1842)

The objective of this study was to compare haematological and serum biochemical parameters of cultured and wild specimens of the northern snakehead, Channa argus, to establish baseline values. Thirty sexually immature and disease‐free wild fish (37.70 ± 13.68 cm total length, 555.3 ± 449.0 g weight) and 30 cultured fish (36.82 ± 1.72 cm total length, 450.5 ± 58.8 g weight) were examined. In cultured northern snakehead, the average values of alanine aminotransferases (370.1 IU L^?1), aspartate amino transferases (1145.3 IU L^?1), albumin (15.84 g L^?1), direct billuribin (6.15 μmol L^?1), urea (1.40 mmol L^?1), glucose (21.54 mmol L^?1) and cholesterol levels (6.60 mmol L^?1) were significantly higher (P < 0.05) than in the wild fish. In wild specimens the corresponding values were 9.81 IU L^?1, 394.1 U L^?1, 12.90 g L^?1, 2.57 μmol L^?1, 0.97, 2.36 and 4.38 mmol L^?1, respectively. No significant difference (P > 0.05) was found for total protein, globulin, total bilirubin, chromium, sodium, chloride or triglyceride levels between wild and cultured populations. The mean values of the red blood cell (RBC) counts, hematocrit, haemoglobin, and mean corpuscular volume (MCV) were significantly higher (P < 0.05) in the cultured population, while the values of the white blood cell (WBC) counts, erythrocyte sedimentation rate (ESR), mean corpuscular haemoglobin (MCH), and mean corpuscular haemoglobin concentration (MCHC) were significantly higher (P < 0.05) in the wild population. The study showed that the environmental conditions significantly impacted the status of the fish. It is suggested that these physiological parameters can be conveniently employed as health monitoring tools in fish culture practices.     

Resting metabolism and heat increment of feeding in mandarin fish (Siniperca chuatsi) and Chinese snakehead (Channa argus)

Resting metabolism was measured in immature mandarin fish Siniperca chuatsi weighing 42.1-510.2 g and Chinese snakehead Channa argus weighing 41.5-510.3 g at 10, 15, 20, 25, 30 and 35 degrees C. Heat increment of feeding was measured in mandarin fish weighing 202.0 (+/-14.0) g and snakehead weighing 200.8 (+/-19.3) g fed swamp loach Misgurnus anguillicaudatus at 1% body weight per day at 28 degrees C. In both species, weight exponent in the power relationship between resting metabolism and body weight was not affected by temperature. The relationship between resting metabolism and temperature could be described by a power function. The temperature exponent was 1.39 in mandarin fish and 2.10 in snakehead (P<0.05), indicating that resting metabolism in snakehead increased with temperature at a faster rate than in mandarin fish. Multiple regression models were used to describe the effects of body weight (W, g) and temperature (T, degrees C) on the resting metabolism (R(s), mg O(2)/h): lnR(s)=-5.343+0.772 lnW+1.387 lnT for the mandarin fish and lnR(s)=-7.863+0.801 lnW+2.104 lnT for the Chinese snakehead. The proportion of food energy channelled to heat increment was 8.7% in mandarin fish and 6.8% in snakehead.     

Food selection and growth of young snakehead Channa striatus

Food selection and growth of young snakehead Channa striatus were studied in the laboratory and in a field trial. In the laboratory, first-feeding snakehead larvae of 6–7 mm total length (TL) with a mouth opening of 0.55 mm selected for Artemia nauplii, and against formulated feed. Fish began feeding on formulated feed at 12mm TL when their mouth width reached 1.0mm. In both laboratory and field trials, snakehead diets changed as fish size increased. For fish 15–20 mm TL, cladocerans and copepods were 96.5% of their diet. With fish 30–40 mm TL, zooplankton consumption was greatly reduced while benthic organism consumption increased. Fish 45–50 mm TL fed exclusively on benthic invertebrates. Diet shift from zoo-plankton to benthic invertebrates was not due to reduced zoo-plankton availability, but was instead related to changes in gill raker structure. Low density of benthic invertebrates in the field trial caused reduced fish growth rates when fish switched diets from zooplankton to benthos. Our results indicate that snake-head can take Artemia nauplii as a larval starter food, then accept formulated feed at ≥12 mm TL. Zooplankton can serve as food for snakehead < 40 mm, but formulated feed should be provided for larger fish which are unable to catch zoo-plankton.     

Distribution of Triassic Foraminifers in the Limestone Massifs of the Dalnegorsk District,Primorsky Krai

Paleontological Journal - Foraminifer assemblages from five massifs of the Triassic limestones of the Dalnegorsk District have been described for the first time, and images of these foraminifers...     

Parasite fauna in salmonidean fishes from the Paanaiarvi-Olanga lake-river system

The data on parasite fauna in the brown trout Salmo trutta, white fish Coregonus lavaretus, and grayling Thymall thymallus from the Paanajarvi-Olanga lake-river system (White Sea basin) are presented. As like-river systems in this area remain in natural conditions, without any anthropogenic pressure, the parasite fauna in salmonoideans from these water bodies had not been changed and has an archetypical state. The parasite fauna core is established in fishes aged 3+. Parasite species forming the core of the parasite fauna infected the brown trout, grayling and whitefish in different degree.     

斑鳢、乌鳢及其杂交种遗传差异的AFLP分析

本文采用AFLP分子标记技术对斑鳢、乌鳢和杂交鳢(斑鳢(母本)与乌鳢(父本))共85个个体(其中斑鳢、乌鳢各30个,杂交鳢25个)进行了遗传差异分析。结果表明11对引物组合共检出了459个不同的扩增片段,扩增出的多态谱带数350条,多态性比例为76.25%,平均每对引物组合扩增出31.8条多态条带。乌鳢与斑鳢种群间存在稳定的、可以简单地借以进行群体鉴别的标记条带169条,其中父本(乌鳢)特异性条带78条,72条能够稳定地遗传给杂交鳢;母本(斑鳢)特异性条带89条,71条能够稳定地遗传给杂交鳢。杂交鳢另外出现了3条非双亲的条带。遗传差异的分子方差分析结果发现,斑鳢与乌鳢种群间的遗传相似度为0.5161,杂交鳢与斑鳢和乌鳢种群间的遗传相似度相近,分别为0.7189和0.7476,斑鳢与乌鳢之间以及杂交鳢与斑鳢和乌鳢之间的种群间遗传距离分别为0.6615、0.3300和0.2909,即AMOVA分析显示斑鳢、乌鳢和杂交鳢间存在极显著的遗传分化。UPGMA聚类分析显示,在个体间,斑鳢与乌鳢能区分成两大类,杂交鳢则分散于斑鳢和乌鳢种群中;在群体间,杂交鳢首先与乌鳢聚类,然后与斑鳢相聚,表明杂交鳢种群总体上更偏向于父本乌鳢。研究结果表明,杂交鳢与斑鳢和乌鳢发生混杂的可能性很大,应该对杂交鳢进行隔离养殖。本文结果将为斑鳢、乌鳢和杂交鳢的遗传分析提供实验依据,也为其种质的合理利用提供参考。     

Hornbeam-broad-leaved-spruce-Korean pine forest in the period from 1926 to 2003 (Ussuriysk Natural Reserve, Southern Primorsky Krai)

Long monitoring (1926–2003) on the permanent test plot laid by B. A. Ivashkevich in the overmature virgin hornbeam-Korean pine forest with spruce shows that the Korean pine is subject to dying-off and has lost its prevailing role in the tree stand. At the same time the tree species dominating the lower layer of tree stand that have completed their life cycle fall off. The dead trees of these species are quickly replaced by young growth, whereas the young generation of Korean pine able to shape a new tree stand has not formed. All this indicates that the cyclicity of the alternation of the Korean pine generations is disturbed. The phase of dominance of deciduous species begins.     

鰤鱼诺卡氏菌对乌鳢血液指标的影响

应用浓度为106cfu/mL的鰤鱼诺卡氏菌(Nocardia seriolea)菌悬液腹腔注射乌鳢,人工感染诺卡氏菌病。在感染后3d、6d、9d、12d、18d和24d时抽取乌鳢血液,检测乌鳢的血细胞数、血细胞脆性、溶菌酶、血清总蛋白、碱性磷酸酶、乳酸脱氢酶和尿素氮等指标。结果表明,与对照组相比,感染鰤鱼诺卡氏菌后乌鳢血细胞数呈先升高后降低的趋势,而血细胞脆性在各实验时间点均高于对照组;血清碱性磷酸酶呈先降低后升高趋势,血清溶菌酶、血清总蛋白和血清尿素氮等均呈先降低后增高再降低的趋势,血清乳酸脱氢酶活力显著增加。说明鰤鱼诺卡氏菌感染会引起乌鳢血液指标明显变化。     

Genetic diversity of a rare species Aristolochia contorta bungae (aristolochiaceae) in Primorsky Krai

The herbaceous vine, twisted birthwort Aristolochia contorta Bunge, is a rare species listed in the Red Data Book of Primorsky krai (2008). On the northern boundary of its range (south of the Russian Far East), the species is represented by small isolated populations confined to the river drainages. Using allozyme analysis, genetic variation of nine natural populations ofA. contorta (247 accessions), which represented the main part of the species range in Russia, was examined. The values of genetic variation indices (P = 22.7%; A = 1.28; Ho = 0.129; He = 0.101) were low and comparable with the data obtained for other rare plants. The proportion of unique genotypes (G/N) and Simpson's genotypic diversity index (D) ranged from 0.32 to 0.64 and from 0.60 to 0.98, respectively. This means that A. contorta is characterized by sexual and asexual reproduction. Moreover, the ratio between these types of reproduction varied among the populations. Complete absence of inbreeding and excess of heterozygotes (F(IS) = -0.282), which was low probable in case of free mating, was observed. Evidently, clonal growth and (or) apomixis enables the species to maintain certain level of heterozygosity despite of small population sizes and non-regular gene exchange.     

Autonomic nervous control of the circulatory system in the air-breathing fish Channa argus

The control of the cardiovascular system with particular emphasis on the regulation of blood distribution in the gills and air-breathing organ was studied in the air-breathing teleost Channa argus. Perfused head preparations were used in addition to experiments with isolated strip preparations of arteries and heart chambers. The distribution of adrenergic nerves was investigated using Falck-Hillarp fluorescence histochemistry. This preliminary study shows an adrenergic control system composed of chromaffin cells and adrenergic nerves similar to that found in other teleosts investigated, although the systemic arteries (coeliac artery, dorsal aorta and the vasculature of the air-breathing organ) appear to lack an adrenergic innervation. The reactions of isolated artery strip preparations to acetylcholine and adrenaline resemble those seen in other teleosts, and there is a prominent inhibitory effect of L-isoprenaline suggestive of arterial beta-adrenoceptors. The general vascular resistance of the gill apparatus-air-breathing organ increases in response to acetylcholine or adrenaline, and there is a redistribution of perfusion flow from the air-breathing organ circuit (anterior venous outflow from the first and second pair of gills and the air-breathing organ) to the general systemic circuit (dorsal aortic outflow from the third and fourth pair of gills). Stimulation of the vagal branch entering the air-breathing organ mimics the effects of acetylcholine or adrenaline. This innervation is probably non-adrenergic since no adrenergic nerve fibres could be demonstrated in the vasculature of the air-breathing organ using the histochemical technique. An adrenergic control of the vasculature of the air-breathing organ is not likely, since the concentration of adrenaline needed to affect the vasculature is not reached in the plasma even during "stress".     

Mitochondrial DNA variation in Olga Bay larch (Larix olgensis A. Henry) from Primorsky Krai of Russia

Two mitochondrial DNA fragments, nad4(3c-4r) and nad5(1-2r), were sequenced in 58 larch accessions from the range of Larix olgensis A. Henry. Combinations of the nad4 polymorphic sites formed four haplotypes, two of which (H3 and H4) were unique and two (H1, H2) were common. Haplotype H1 was found only in pure L. olgensis from the vicinity of Olga Bay and in a number of accessions from the southern part of the range. Haplotype H2 was detected in the other samples from the range of Olga Bay larch, as well as in hybrid forms. Similarly to the nad4(3c-4r) fragment, the mtDNA fragment UBC460 was able to differentiate larch populations from the range of L. olgensis examined.     

Electron microscopic studies on the retinal photoreceptors of the spotted snakehead, Channa punctatus

The retinal photoreceptors of Channa punctatus include rods, single cones and the double cones. The double cones have the outer segments having lamellae which are distorted into tubules and vesicles, and the inner segments with hypertrophied mitochondria.     

A new record ofAchlya klebsiana from snakehead,Channa striatus,with fungal infection in Myanmar

This is the first record of the discovery in Myanmar ofAchlya klebsiana, belonging to the family Saprolegniaceae.     

An Angiocardiographic Analysis of the Central Circulation in the Air Breathing Teleost Channa argus

The unique anatomy of the double ventral aorta outflow system in the air breathing teleost Channa argus (Ophiocephalus) showing an anterior and posterior ventral aorta is described. The marked trabeculation of the ventricle and bulbus arteriosus and the arrangement of central veins are used as a basis for the hypothesis that Channa may selectively channel the well oxygenated blood draining the air breathing organs via the anterior cardinal vein to the posterior ventral aorta, which forms the systemic arterial circulation. An angiocardiographic technique was used to test this hypothesis, as well as to delineate the functional role of the heart chambers in the cardiac cycle. No reflux of contrast to the sinus venosus during atrial filling and no ventricular filling before atrial contraction were apparent, which makes the atrium the main determinant of the ventricular end-diastolic volume. Ventricular contraction left a small or no residual volume. The ventricular ejectate was initially nearly completely absorbed by the very elastic bulbus arteriosus, acting as a pressure chamber (Windkessel) stabilizing and prolonging ventral aortic blood flow. Contrast medium was not selectively passed from the anterior cardinal vein to the posterior ventral aorta. However, the diameter of this vessel and its density of contrast were greater than in the anterior aorta, suggesting a preference for a greater blood flow from the air breathing organ through the heart to the posterior aorta.     

Invertebrate drift in the piedmont part of the Kedrovaya River (Primorsky Krai,Russia) in warm season

Diurnal dynamics of invertebrate drift in the Kedrovaya River (Primorsky Krai, Russia), which flows in the zone of influence of the monsoon climate, has been studied. It has been shown that drifting invertebrates tend to shift from a predominantly daytime drift pattern to a distinguished nocturnal drift pattern during the frost-free period. The ratio of the total number of nighttime migrants to the total number of daytime migrants increased with every subsequent month. It has been proposed that the degree of light contrast between day and night can be significant in the regulation of nocturnal drift intensity.     

Parasite fauna of the amur sickleback Pungitius sinensis ( Gasterosteidae) from the Primor'e region

An ecological analysis of parasite fauna occurring in the Amur sickleback in different water basins of the Maritime Territory is carried out. Possible pathways of distribution of this fish within the Maritime Territory is discussed.