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。     

Origin and Development of Neutrophils in an Air Breathing Fish,Channa punctatus Bloch

Mahajan, C. L., Dheer, J. M. S. 1980. Origin and development of neutrophils in an air breathing fish, Channa punctatus Bloch. (Department of Zoology, University of Rajasthan, India.) — Acta zool. (Stockh.) 61(4): 221–224. An account of the study of the development of neutrophils based on detailed examinations of kidney and spleen imprints has been given in this paper. Each developmental stage has been investigated with respect to its morphology, morphometry and cytochemistry using certain special techniques developed in this laboratory for fishes such as differential haemoglobin staining, autoradiography etc. Five main stages (large lymphoid haemoblasts, promyelocytes, mesomyelocytes, metamyelocytes, mature neutrophils) have been identified in development and are described in detail.     

Parasite fauna of the snakehead Channa argus warpachowskii from Primorsky Krai

Parasite fauna of the snakehead Channa argus warpachowskii was studied in water bodies of Primorsky Krai. Thirty-one parasite species were recorded in this host from the studied area according to original and literary data. Most part of these species are host-specific parasites of snakehead originated from Chinese-Indian region or parasites having a wide host range and able to infest almost all freshwater fishes. Moreover, three brackishwater species of parasites were found in the fishes introduced into the rivers of the southern part of Primorsky Krai. The causes of this invasion are discussed.     

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".     

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。     

乌鳢三倍体诱导及其生长

采用热休克抑制第二极体排放的方法诱导乌鳢(Channa argus)三倍体,以探索人工诱导乌鳢三倍体的理想热休克条件。采用DNA含量测定法和红细胞核大小鉴定法对获得的鱼苗倍性进行鉴定,同时对普通二倍体乌鳢群体与三倍体群体的生长差异进行了比较研究。结果表明,(1)热休克法适宜诱导条件为,(28±0.5)℃培育水温授精后4 min,在水温42℃条件下持续处理3 min,三倍体诱导率最高,达到87.69%;(2)三倍体与二倍体DNA含量差异极显著(P0.01),其比值为1.50︰1.00;(3)三倍体和二倍体在红细胞核长径、红细胞核体积、红细胞核面积等6项指标上存在极显著差异(P0.01);与二倍体相比,三倍体红细胞体积和核体积分别是其1.69倍和1.60倍;(4)在4月龄与8月龄,三倍体的体长和体重比二倍体稍高,但两者差异不显著(P0.05)。本实验结果为进一步开展乌鳢倍性育种奠定了基础。     

Effect of meal size on the specific dynamic action of the juvenile snakehead (Channa argus)

The effect of meal size on the specific dynamic action (SDA) of the juvenile snakehead (Channa argus) was assessed at 25 °C. The fish were fed with test diets at meal sizes of 0.5, 1, 2, 3, 4, and 5% body mass and the postprandial oxygen consumption rate was determined at 1-h intervals until it returned to the pre-prandial level. The peak metabolic rate increased from 237.4 to 283.2 mg O₂ kg^− 1 h^− 1 as the relative meal size increased from 0.5% to 3% and leveled off at 4% and 5%. Factorial metabolic scope increased from 1.53 to 1.99 and SDA duration increased from 11.7 to 32.3 h as the relative meal size increased from 0.5% to 5%. The relationship between SDA duration (D) and relative meal size (M) was described as: D = 4.28 M + 10.62 (r² = 0.752, P < 0.05, n = 50). The energy expended on SDA increased while the SDA coefficient decreased with increasing meal size. The results of the present study suggest that the snakehead may adopt different feeding strategies when taking in different amounts of food.     

The Cerebral Cost of Breathing: An fMRI Case-Study in Congenital Central Hypoventilation Syndrome

Certain motor activities - like walking or breathing - present the interesting property of proceeding either automatically or under voluntary control. In the case of breathing, brainstem structures located in the medulla are in charge of the automatic mode, whereas cortico-subcortical brain networks - including various frontal lobe areas - subtend the voluntary mode. We speculated that the involvement of cortical activity during voluntary breathing could impact both on the “resting state” pattern of cortical-subcortical connectivity, and on the recruitment of executive functions mediated by the frontal lobe. In order to test this prediction we explored a patient suffering from central congenital hypoventilation syndrome (CCHS), a very rare developmental condition secondary to brainstem dysfunction. Typically, CCHS patients demonstrate efficient cortically-controlled breathing while awake, but require mechanically-assisted ventilation during sleep to overcome the inability of brainstem structures to mediate automatic breathing. We used simultaneous EEG-fMRI recordings to compare patterns of brain activity between these two types of ventilation during wakefulness. As compared with spontaneous breathing (SB), mechanical ventilation (MV) restored the default mode network (DMN) associated with self-consciousness, mind-wandering, creativity and introspection in healthy subjects. SB on the other hand resulted in a specific increase of functional connectivity between brainstem and frontal lobe. Behaviorally, the patient was more efficient in cognitive tasks requiring executive control during MV than during SB, in agreement with her subjective reports in everyday life. Taken together our results provide insight into the cognitive and neural costs of spontaneous breathing in one CCHS patient, and suggest that MV during waking periods may free up frontal lobe resources, and make them available for cognitive recruitment. More generally, this study reveals how the active maintenance of cortical control over a continuous motor activity impacts on brain functioning and cognition.     

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.     

Seasonal and Daily Variations in Hypothalamic Monoamine Levels and Monoamine Oxidase Activity in the Teleost Channa Punctatus (Bloch)

In Channa punctatus, a significant daily variation in hypothalamic S-HT level and monoamine oxidase (MAO) activity was noticed in preparatory phase (February), but not in prespawning (May) or postspawning (November) phases. Hypothalamic dopamine (DA) and noradrenaline (NA), on the other hand, showed marked daily variation in their levels during all the three seasons with peak values in the photophase. The overall activity of MAO (mean ± SEM on 24-hr period) increased from November to May through February, whereas the 5-HT content which was high in November decreased during February and May. The NA and DA levels were low in November and Februry and high in May. The catecholamine (CA) content and MAO activity increased with increasing photoperiod and temperature which is indicative of an enhanced CA metabolism.     

Seasonal and Daily Variations in Hypothalamic Monoamine Levels and Monoamine Oxidase Activity in the Teleost Channa Punctatus (Bloch)

In Channa punctatus, a significant daily variation in hypothalamic S-HT level and monoamine oxidase (MAO) activity was noticed in preparatory phase (February), but not in prespawning (May) or postspawning (November) phases. Hypothalamic dopamine (DA) and noradrenaline (NA), on the other hand, showed marked daily variation in their levels during all the three seasons with peak values in the photophase. The overall activity of MAO (mean ± SEM on 24-hr period) increased from November to May through February, whereas the 5-HT content which was high in November decreased during February and May. The NA and DA levels were low in November and Februry and high in May. The catecholamine (CA) content and MAO activity increased with increasing photoperiod and temperature which is indicative of an enhanced CA metabolism.     

Episodic Breathing in Frogs: Converging Hypotheses on Neural Control of Respiration in Air Breathing Vertebrates

SYNOPSIS. The episodic, or intermittent, breathing of frogsand many ectothermic vertebrates results in important fluctuationsof arterial blood gases. This pattern of breathing differs fromthe rhythmic and continuous alternation of inspiration observedin most homeotherms, which maintain O₂ and CO₂ levels withinnarrow ranges. These differences in pattern of breathing indicatethat the respiratory control systems of ectotherms and homeothermsdiffer substantially. The results of recent studies using invitro brainstemspinal cord preparations of adult frogs and premetamorphictadpoles (Rana catesbeiana and Rana pipiens) demonstrate, however,that the mechanisms for rhythm generation and pattern formationdescribed previously for mammals are also key features of therespiratory control system of frogs. These findings thereforesupport the hypothesis that the respiratory control system ishighly conserved amongst air breathing vertebrates, whetherthey breathe continuously or episodically.     

An Evolutionary Perspective for Bimodal Respiration: A Biological Synthesis of Fish Air Breathing

SYNOPSIS: The broad phyletic distribution of bimodal breathingamong the fishes, the diversity of this group's aerial-respiratoryspecializations, and the numerous ways that bimodality has permeatedthe natural history of many species provide a broad perspectiveon the evolution and biological significance of bimodal breathing.The long term evolutionary view of bimodal breathing emphasizesits importance in the evolutionary transition to terrestriality;a key role in this process was played by the fishes which werethe first air-breathing vertebrates. On the other hand, in mostextant fishes bimodality has not led to terrestriality. Rather,auxiliary air breathing enables a species to remain in or toexploit an aquatic habitat from which it would otherwise beexcluded.     

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.     

Respiratory Control in the Transition from Water to Air Breathing in Vertebrates

SYNOPSIS. Studies on extant bimodally breathing vertebratesoffer us a chance to gain insight into the changes in respiratorycontrol during the evolutionary transition from water to airbreathing. In primitive Actinopterygian air-breathingfishes(Lepisosteus and Amid), gill ventilation is driven by an endogenouslyactive central rhythm generator that is powerfully modulatedby afferent input from internally and externally oriented branchialchemoreceptors, as it is in water-breathing Actinopterygians.The effects of internal or external chemoreceptor stimulationon water and air breathing vary substantially in these aquaticair breathers, suggesting that their roles are evolutionarilymalleable. Air breathing in these bimodal breathers usuallyoccurs as single breaths taken at irregular intervals and isan on-demand phenomenon activated primarily by afferent inputfrom the branchial chemoreceptors. There is no evidence forcentral CO₂/pH sensitive chemoreceptors and air-breathing organmechanoreceptors have little influence over branchial- or air-breathingpatterns in Actinopterygian air breathers. In the Sarcopterygianlungfish Lepidosiren and Protopterus, ventilation of the highlyreduced gills is relatively unresponsive to chemoreceptor ormechanoreceptor input. The branchial chemoreceptors of the anteriorarches appear to monitor arterialized blood, while chemoreceptorsin the posterior arches may monitor venous blood. Lungfish respondvigorously to hypercapnia, but it is not known whether theseresponses are mediated by central or peripheral chemoreceptors.A major difference between the Sarcopterygian and Actinopterygianbimodal breathers is that lungfish can inflate their lungs usingrhythmic bouts of air breathing, and lung mechanoreceptors influencethe onset and termination of these lung inflation cycles. Thecontrol of breathing in amphibians appears similar to that oflungfish. Branchial ventilation may persist as rhythmic buccaloscillations in most adults, and stimulation of peripheral chemoreceptorsin the aortic arch or carotid labyrinths initiates short boutsof breathing. Ventilation is much more responsive to hypercapniain adult amphibians than in Actinopterygian fishes because ofcentral CO₂/pH sensitive chemoreceptors that act to convertperiodic to more continuous breathing patterns when stimulated.     

Structure and composition of teleost scales from snakehead Channa argus (Cantor) (Perciformes: Channidae)

The structure of teleost scales from snakehead Channa argus was investigated using thermogravimetric analysis (TG), scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy-dispersive analysis of X-rays (EDAX), Fourier transform infra-red spectroscopy (FTIR) and X-ray diffraction (XRD). Thermal treatment of fish scales indicates that the fibrillary plate is partially calcified. SEM shows two kinds of scale denticles, arranged along the circuli in the anterior field and the lateral fields, respectively. TEM indicates the stratum laxum with abundant fibrils, chromatophores and capillary blood vessels within the scale covering, and shows the fibrillary plate as an 'orthogonal plywood structure' of stratified lamellae, consisting of 80–100 nm diameter collagen fibres co-aligned in individual lamellae and alternated by c. 90° of the fibre alignment between adjacent lamellae. EDAX, FTIR and XRD show that the mineral phase of the scales is a carbonated hydroxyapatite with a Ca:P molar ratio of 1·85.     

Properties of Respiratory Pigments in Bimodal Breathing Animals: Air and Water Breathing by Fish and Crustaceans

SYNOPSIS. Oxygen is nearly 30 times more available in air comparedwith water, whereas the carbon dioxide capacity of water is{small tilde}28.5-fold greater than for oxygen, presenting bimodalbreathing species with two very different respiratory milieus.The respiratory pigment plays a variable role in animals switchingbetween the two media. In vertebrates the transition to airbreathing involves two main strategies: a decrease in oxygenaffinity and changes in other haematological parameters suchas haematocrit. When appropriately analyzed, data reveal a decreasein blood oxygen affinity during transition to air. This mayarise via differences in the intrinsic affinity as occurs insome amphibians, or be due to increasesin the organic phosphate:haemoglobin ratio when acclimating to air breathing. Adoptingair breathing often promotes increased haematocrit. It is difficultto discern trends in haemocyanin functioning. Many but not allbimodal and air breathingspecies of crab contain haemocyaninwith high affinity for oxygen. As with haemoglobin there issome tendency for blood haemocyanin concentration to increasewith air breathing but bimodal species are quite variable inthis regard. Different strategies for breathing air are employedby various bimodal crustaceans, some of which involve modulationof haemocyanin oxygenaffinity. The exact mechanisms are oftenspecies dependent and in all bimodal breathing organisms therole of the pigment is best appreciated when the demands ofthe local environment and the behaviour of the species are considered.