A rapid onset, post-capture muscle necrosis in the Norway lobster, Nephrops norvegicus (L.) from the West coast of Scotland

A post-capture, abdominal muscle necrosis of rapid onset has been identified in Norway lobsters, Nephrops norvegicus (L.), captured off the West coast of Scotland. Economic losses, as a result of the mortality of these animals in transport, were encountered by Scottish wholesalers during the summer and autumn of 1999. Affected animals show a characteristic whitening of individual muscle fibres and fibre bundles of the abdomen within hours of capture, with a progression towards complete opacity of the abdominal musculature within a number of days. The pathology causes a loss of the normal function of the abdomen; thus, preventing the normal 'tail flip' swimming. Electron microscopy failed to reveal any obvious causative agent but showed that affected tissue displayed a progressive disruption of sarcomere organization, loss of Z-line material, condensation of myofibrils and infiltration of necrotic regions by granulocytes. SDS–PAGE of affected muscle tissue showed that there was a great reduction of most of the major contractile proteins. The condition most closely resembles idiopathic or spontaneous muscle necrosis, a pathology previously reported from both wild and cultured crustaceans. Damage to the integument in conjunction with exposure to various stressors during and immediately after capture is the most likely cause of the pathology. The rapid onset of the pathology has implications for the post-capture handling procedure for N. norvegicus and their subsequent vivier transport to market. It may also be partially responsible for the high mortality rate of undersized N. norvegicus returned to the sea after capture and aerial emersion.     

氨和空气暴露对大鳞副泥鳅组织中谷氨酰胺含量的影响

为研究氨对大鳞副泥鳅(Paramisgurnus dabryanus)(体重18~25 g)生理指标的影响,将其暴露于30 mmol/L NH_4Cl溶液和空气中0 h,6 h,12 h,24 h,48 h,72 h,以研究体外和体内氨对其体组织中谷氨酰胺含量、谷氨酰胺合成酶和谷氨酸脱氢酶活性的影响。研究发现,氨和空气暴露下,随着暴露时间的延长,大鳞副泥鳅肝脏和肌肉组织中谷氨酰胺含量有明显累积的趋势,脑、肝脏和肠道组织中谷氨酰胺合成酶活性均显著上升。氨和空气暴露显著影响大鳞副泥鳅脑和肠道组织中谷氨酸脱氢酶活性,但对肝脏组织中谷氨酸脱氢酶活性并没有显著性影响。结果表明,大鳞副泥鳅可通过体组织中累积谷氨酰胺来应对体外或体内氨浓度的上升,氨暴露和空气暴露可刺激体内谷氨酰胺的合成,将氨转化为无毒性的谷氨酰胺。肠道中谷氨酸脱氢酶活性显著上升,可能在鱼类应对氨氮毒性中,肠道谷氨酸脱氢酶比谷氨酰胺合成酶有更重要的作用。而大鳞副泥鳅肝脏组织中谷氨酸脱氢酶活性并不受氨和空气暴露的影响,这可能是由于肝脏组织中转氨酶催化生成了足量的谷氨酸。     

Co‐Infection with Pseudomonas anguilliseptica and Delftia acidovorans in the European eel,Anguilla anguilla (L.): a case history of an illegally trafficked protected species

Inspections by customs agents at Barcelona airport discovered 420 kg of contraband glass eels prepared for shipment to Hong Kong. After confiscation of these animals by police, they were transported to holding facilities to be maintained until after a judicial hearing. Upon arrival, they were separated into two groups and held under ambient flow‐through conditions in fresh water. During their captivity period, several peaks in mortality occurred and multiple bacterial strains were isolated from moribund animals. Sequencing of 16S rDNA was used to determine specific identity of the isolates. An initial isolation of Pseudomonas anguilliseptica was treated with oxytetracycline. A subsequent isolation of Delftia acidovorans proved resistant to oxytetracycline and was treated with gentamicin in combination with sulphadiazine–trimethoprim. Once the health condition of the animals was stabilized, they were partitioned into groups and subsequently released as part of a restocking effort for the species following the guidelines of Regulation (EC) 1100/2007 (Anon 2007). This represents the first record for both bacterial species in the host Anguilla anguilla in the Spanish Mediterranean.     

Quantitative analysis of an experimental white spot syndrome virus (WSSV) infection in Penaeus monodon Fabricius using competitive polymerase chain reaction

White spot syndrome virus (WSSV) has been a major pathogen of cultured Penaeus monodon Fabricius in Malaysia since 1994. As quantitative study on the replication of WSSV is in its infancy, competitive polymerase chain reaction (PCR) was used for quantitative study of an experimental WSSV infection per os in growout P. monodon . Gills, abdominal integument and abdominal muscle were selected for viral quantification. Infection was detectable as early as 14 h postinfection (h p.i.) in both gills and integument, but the infection in muscle was only detected at 24 h p.i. Gill tissue had the highest viral load, followed by integument and muscle. Typical viral growth curves were obtained for all organs with distinct phases of eclipse (0–24 h p.i.), logarithmic (24–48 h p.i.) and the plateau (48–120 h p.i.). Cumulative mortality rapidly increased from 48 h p.i. and reached 100% at the end of the plateau phase at 120 h p.i. Gross signs of white spots and reddish discoloration were also obvious in moribund individuals from the plateau phase. Based on the three phases of viral growth, WSSV infection was classified into light, moderate and heavy infection stages.     

Evaluation of the link between gyrodactylosis and streptococcosis of Nile tilapia, Oreochromis niloticus (L.)

Streptococcus iniae and Gyrodactylus niloticus are two common pathogens of cultured Nile tilapia, Oreochromis niloticus. We studied concurrent infection of tilapia by G. niloticus and S. iniae and evaluated whether parasitism in tilapia with Gyrodactylus increased susceptibility and mortality following immersion infection with S. iniae. Results showed that death mainly occurred in fish with G. niloticus and challenged with S. iniae (G-S group). The accumulative mortality (42.2%) was significantly higher in the G-S group than in fish not infected by the parasite (6.7%), but exposed to S. iniae. Bacteriological examination revealed S. iniae from > or =92% of dead or moribund fish challenged with S. iniae. Gyrodactylus not only damaged fish epithelium and provided entry for invasive bacteria but also was found to harbour viable cells of S. iniae for 24 and 72 h. Streptococcus iniae was isolated from 60% and 40% of G. niloticus collected from fish infected by intraperitoneal injection or immersion, respectively, at 24 h post-challenge. The present study confirms that parasitism of tilapia by G. niloticus increased host mortality following exposure to the bacterial pathogen S. iniae.     

Routes of water loss in South African abalone (Haliotis midae) during aerial exposure

South African abalone, Haliotis midae, were exposed to air at 12 °C for 36 h to simulate the extent and rate mass loss experienced by animals during long distance live exports. Animals lost 15.1 ± 0.94% of their mass during the 36 h air exposure, an approximation of the highest mass losses sustained by industry.The total mass loss was attributed to water loss, as the contribution of dry mass to the total mass remained constant under all conditions. Water content decreased from 64.8% of the body mass (M_b) under control conditions to 58.8% M_b after 36 h in air. In real terms, however, animals had lost 22% of the body water pool.Abalone exhibited a typically high water turnover rate when in water (125 μL g^− 1 h^− 1), which decreased markedly during air exposure (2.2 μL g^− 1 h^− 1). Haemolymph volume decreased from 43% M_b in water to 14% M_b in air. The concomitant decrease in haemolymph pressure probably limited the first step in urine formation (ultra-filtration through the pericardium). Thus we observed that while urine flow represented about 26% of the total water loss when the animals were in water, urine flow ceased during air exposure.The decrease in haemolymph volume in air represents a redistribution of water to the tissues and not a bulk loss of haemolymph. This is supported by the concentration of haemolymph ions by a factor of 1.2 during aerial exposure, which was predicted based on the 22% decrease in water content. Under the same conditions, evaporation from water containers with similar surface to volume dimensions as abalone, accounted for only an 8.25% mass loss. As all other water loss routes were accounted for, we measured pedal mucus production rates of abalone in water and air. During 36 h aerial exposure, the pedal mucus production represented a loss of 6.8% M_b. We conclude that water loss during 36 h air exposure is attributable to evaporation (8.25% M_b) and pedal mucus production (6.8% M_b). This paves the way for directed research into mitigating water loss during the live export process.     

Evidence of enhanced bacterial invasion during Diplostomum spathaceum infection in European grayling, Thymallus thymallus (L.)

Farmed grayling, Thymallus thymallus (L.), are susceptible to atypical Aeromonas salmonicida (aAS) infections. Interactions between bacteria and parasites were studied using grayling subjected to concomitant exposure to aAS bacteria and the digenean parasite Diplostomum spathaceum. Atypical AS was detected from fish by a combination of bacterial cultivation and polymerase chain reaction techniques. A detection level of 17 aAS cells per 100 mg intestine tissue sample was obtained. Concomitant bacterial exposure did not enhance the severity of grayling eye rupture and nuclear extrusion induced by D. spathaceum, but D. spathaceum invasion into grayling increased the proportion of fish carrying aAS in their heart tissue. However, the number of aAS cells detected in heart tissue was low. Atypical AS did not cause acute disease or mortality during 15 days post-exposure. There was a higher prevalence of aAS in grayling heart samples than in intestinal samples, indicating that the intestine is not favoured by aAS. We suggest that heart tissue would be a good organ from which to isolate aAS when tracing latent carrier fish. We conclude that penetrating diplostomids can enhance bacterial infections in fish and that diplostomids can cause serious eye ruptures in grayling.     

Quantitative real time PCR for the measurement of white spot syndrome virus in shrimp

Quantitative real time PCR, recently developed in molecular biology, is applied in this paper to quantify the white spot syndrome virus (WSSV) in infected shrimp tissue. The WSSV content in moribund shrimp of all species tested ( Penaeus stylirostris, P. monodon, P. vannamei ) ranged from 2.0 × 10⁴ to 9.0 × 10¹⁰ WSSV copies μg^–1 of total DNA ( n =26). In whole moribund post-larvae, 4.3 × 10⁹ WSSV copies μg^–1 of DNA were detected which is equivalent to 5.7 × 10¹⁰ WSSV copies g^–1 of post-larvae. The comparison of WSSV content between different tissues showed that muscle and hepatopancreas tissues contained 10 times less virus than gills, pleopods and haemolymph. With inocula of known virus content, bioassays by immersion challenge showed that a minimum of five logs of WSSV copies was necessary to establish disease in the challenged shrimp. In contrast, five logs of WSSV copies injected into shrimp muscle produced a LT-50 of 52 h. This real time polymerase chain reaction (PCR) technique is sensitive (four copies), specific (negative with DNA from shrimp baculoviruses and parvoviruses), dynamic (seven logs) and easy to perform (96 tests in <4 h).     

The pathology of chronic erosive dermatopathy in Murray cod, Maccullochella peelii peelii (Mitchell)

Chronic erosive dermatopathy (CED) is a disease of intensively farmed Murray cod in Australia that has been reported in association with the use of groundwater (mechanically extracted from shallow boreholes) supplies. CED results in focal ulceration of the skin overlying sensory canals of the head and flanks. Trials were conducted at an affected fish farm to study the development of the condition, both in Murray cod and in goldfish, and also to assess the reported recovery of lesions when affected fish were transferred to river water. Grossly, lesions began after 2-3 weeks with degeneration of tissue at the periphery of pores communicating with the sensory canals. Widening of these pores along the axis of the canals resulted from a loss of tissue covering the canal. Histopathologically, hyperplasia of the canal epithelial lining was seen after 3 weeks in borehole water and subsequent necrosis and sloughing of this tissue resulted in the loss of the canal roof. Canal regeneration occurred when fish were transferred from borehole water into river water. The lack of lesions in other organs and the pattern of lesion development support exposure to waterborne factors as the most likely aetiology.     

A method for inducing muscle relaxation in the abalone, Haliotis iris

Four chemical and two non-chemical methods for induction of relaxation in the abalone, Haliotis iris, were tested for prospective use in pearl seeding. Requirements were for rapid onset of relaxation, full extension and softening of the foot and epipodium muscle for at least 10 min, and recovery with minimal stress and no mortality. Nembutal at 1 ml/l was most successful, producing relaxed and soft muscle with good accessibility to operation sites, and no mortality. Benzocaine and MS-222 made animals release from container walls within 30–60 min, but the animal's muscle was hard and contracted. Propylene phenoxytol caused muscle contraction and mortality at 2.5 ml/l. The effects of the chemical relaxants were similar in warm (21–22.5°C) and cool (15–16°C) water. Elevation of the water temperature from 18°C to 23°C (without chemicals) caused some softening of the muscle, but was much less effective than Nembutal. Air exposure for up to 4 h was ineffective. This work provides a simple method to relax live abalone for pearl nucleation or other manipulations.     

Uptake and Clearance of Edwardsiella ictaluri in the Peripheral Blood of Channel Catfish Ictalurus punctatus Fingerlings during Immersion Challenge

Uptake and clearance of Edwardsiella ictaluri in the peripheral blood of channel catfish Ictalurus punctatus fingerlings were monitored for 216 h after exposure to E. ictaluri for 4 h and 8 h under static conditions. Most fish exposed to E. ictaluri developed bacteriemia 24 h post-exposure, and by 72 h post-exposure E. ictaluri was recovered from all the blood of all exposed fish. The number of E. ictaluri colony forming units (CFU) in the blood of moribund fish ranged between 1.7 × 10³ to 1.6 × 10⁵ CFU/50 μL whole blood. Clearance of bacteria from the blood was observed by 216 h post-exposure and all fish surviving bacterial exposure developed agglutinating antibody against E. ictaluri . The pathogenesis of the infection was accompanied by the shedding of viable E. ictaluri into the water which may serve as a mechanism by which fish to fish transmission occurs.     

Susceptibility of channel catfish, Ictalurus punctatus (Rafinesque), to Edwardsiella ictaluri challenge following copper sulphate exposure

Channel catfish, Ictalurus punctatus (Rafinesque), with or without a preliminary 24 h exposure to 2 mg copper sulphate L(-1), were challenged with 7.5 x 10(6) colony forming units L(-1) of Edwardsiella ictaluri to determine the effect of copper sulphate on disease resistance. Catfish previously exposed to copper sulphate were significantly more resistant to the bacterial challenge than those not exposed. Catfish not exposed to copper sulphate suffered 35.5% mortality while catfish exposed to copper sulphate experienced 14.1% mortality. Copper concentrations were the same in tank waters of both exposed and control fish at the time of challenge, eliminating the possibility that copper in the water may have been toxic to bacteria. Copper concentrations in freeze dried and ground tissues of unexposed, exposed, and purged channel catfish were highest in fish before copper sulphate exposures suggesting that elevated tissue levels of copper were not responsible for the increased resistance to bacterial challenge. Competition for sites of bacterial attachment to gill or epithelial cells may account for the reduction in mortality; although this is not supported by the low copper content of fish tissue after copper exposure.     

养殖鲟鱼暴发病病原菌分离及药敏实验

从濒死的西伯利亚鲟(Acipenser baerii)心、肝、肾、脾等组织中分离到细菌性病原20株,并对分离的菌株进行了细菌生化编码鉴定管鉴定及药敏实验。结果表明,分离株分别包括致伤弧菌(Vibrio vulnificus)6株、豚鼠气单胞菌(Aeromonas caviac)5株、温和气单胞菌(Aeromonas sobria)5株、亲水气单胞菌(Aeromonas hydrophila)4株。16种常见药物的药敏实验显示,分离菌株均对多粘菌素B、妥布青霉素等6种药物较敏感;对红霉素、四环素等6种药物耐药,仅有部分菌株表现出不同程度的耐药性及敏感度的变化趋势。该研究对于合理使用抗菌药物,防治养殖鲟细菌性疾病及其他疾病引起的细菌继发性感染提供一定的数据基础。     

氨和空气暴露对大鳞副泥鳅肝脏抗氧化能力的影响

将大鳞副泥鳅(Paramisgurnus dabryanus)分别暴露于 30 mmol/L NH₄Cl 溶液和空气中, 以评价氨和空气暴露对其肝脏抗氧化能力的影响。结果显示, 氨暴露并未引起大鳞副泥鳅肝脏超氧化物歧化酶(superoxide dismutase, SOD)活性的显著变化, 仅空气暴露 12 h 后, 大鳞副泥鳅肝脏 SOD 活性显著高于对照组(P<0.05)。氨和空气暴露均显著降低了大鳞副泥鳅肝脏过氧化氢酶 (catalase, CAT) 活性 , 且在暴露 72 h 后升高 (P<0.05) 。丙二醛 (malondialdehyde, MDA)及脂质过氧化物(lipid peroxide, LPO)含量在氨和空气暴露组中均呈现先显著上升后下降的趋势。氨和空气暴露对大鳞副泥鳅肝脏谷胱甘肽-S 转移酶(glutathione S-transferase, GST)活性、谷胱甘肽(glutathione, GSH)含量和总抗氧化能力(total antioxidant capacity, T-AOC)无显著影响(P>0.05), 但谷胱甘肽过氧化物酶 (glutathione peroxidase, GSH-Px)活性均明显降低, 乙酰胆碱酯酶(acetylcholin esterase, AChE)活性均显著上升。综上所述, 在氨和空气暴露的初期, 大鳞副泥鳅体内出现了明显的氧化反应; 而在暴露一段时间后, 体内的氧化反应受到了明显的抑制, 大鳞副泥鳅肝脏中 SOD、CAT、GST 及 GSH-Px 等抗氧化酶系统并未被成功激活。     

Histopathological effects of cadmium on the gills of the freshwater fish, Macropsobrycon uruguayanae Eigenmann (Pisces, Atherinidae)

Evaluations of histopathological lesions in gill tissue were carried out in the freshwater fish Macropsobrycon uruguayanae following 30 and 60 days of exposure to 1.5 mg 1^-1 of cadmium. The study was conducted on both fed and starved animals in order to determine the influence of feeding condition on cadmium toxicity. The main lesions observed and quantified were: (1) hyperplasia of primary lamellar epithelium; (2) hyperplasia of secondary lamellar epithelium; (3) separation of respiratory epithelium; (4) shortening of secondary lamellae; (5) epithelial necrosis; (6) fusion of adjacent secondary lamellae; (7) hypertrophy of respiratory epithelium; (8) lamellar telangiectasis; (9) hyperplasia of chloride cells; (10) mucinous metaplasia; and (11) inflammatory infiltration. Lesions 6, 8, 9 and 11 were only induced by exposure to cadmium, while lesion 4 could be produced only by starvation. Starved fish also showed a reduction in total body weight and length. Lesion 2 was shown to be non-specific, and produced by either cadmium, starvation or even exposure time. The possible mode of action of the experimental factors are discussed in relation to the observed pathologies.     

干露胁迫对脊尾白虾(Exopalaemon carinicauda)呼吸代谢相关酶的影响

为了研究离水干露对脊尾白虾(Exopalaemon carinicauda)呼吸代谢能力的影响,以常温干露(22-24℃)、低温干露(4℃)2个实验组和1个对照组进行脊尾白虾干露胁迫实验,于胁迫后60、90、120、180 min取组织,测定琥珀酸脱氢酶(SDH)、细胞色素氧化酶(CCO)、延胡索酸还原酶(FRD)、乳酸脱氢酶(LDH)4种呼吸代谢酶活力和乳酸含量.结果显示,常温干露组肝胰腺、鳃、肌肉SDH和CCO活力随干露时间的延长而降低,且显著低于对照组(P＜0.05);FRD、LDH活力和肌肉乳酸含量随胁迫时间的延长呈先升高后降低趋势,且显著高于对照组(P＜0.05).低温干露组在胁迫60 min时,各组织呼吸代谢酶活力和肌肉乳酸含量与对照组无显著差异(P＞0.05),之后的变化趋势和常温干露组相同.常温干露组和低温干露组中肝胰腺和鳃SDH/FRD比值的变化幅度大于肌肉.研究表明,脊尾白虾通过提高无氧代谢来应对干露胁迫;低温干露下的脊尾白虾在胁迫早期可以维持有氧呼吸,肝胰腺和鳃比肌肉对干露胁迫更加敏感.     

Recovery of yellow-pigmented bacteria from dead and moribund fish during outbreaks of rainbow trout, Oncorhynchus mykiss (Walbaum), fry syndrome in England

Abstract. During Autumn 1990, the possible presence of microbial pathogens was investigated in two discrete populations of anaemic (rainbow trout fry syndrome; RTFS) rainbow trout fry. Dense pure culture growth of three types of yellow pigmentcd bacteria were recovered from moribund and dead animals on low nutrient media. The cultures were identified as coryneforms. Cytophaga columnaris and Janthinobacterium sp. All taxa induced clinical disease (with similarities to RTFS) in rainbow trout fry.     

Adaptations of amphibious fish for surviving life out of water

There are a small number of fish species, both marine and freshwater, that exhibit a truly amphibious habit that includes periods of aerial exposure. The duration of emersion is reflected in the level of physical and physiological adaptation to an amphibious lifestyle. Fish that are only briefly out of water retain predominantly aquatic attributes whereas there are semi‐terrestrial species that are highly adapted to prolonged periods in the aerial habitat. Desiccation is the main stressor for amphibious fish and it cannot be prevented by physiological means. Instead, amphibious fish resist excessive water loss by means of cutaneous modification and behavioural response. The more terrestrially adapted fish species can tolerate considerable water loss and may employ evaporation to aid thermoregulation. The amphibious habit is limited to fish species that can respire aerially. Aerial respiration is usually achieved through modification to existing aquatic pathways. Freshwater air‐breathers may respire via the skin or gills but some also have specialized branchial diverticula. Marine species utilize a range of adaptations that may include modified gills, specialized buccopharyngeal epithelia, the intestine and the skin. Areas of enhanced respiratory activity are typified by increased vascularization that permits enhanced perfusion during aerial exposure. As with other adaptations the mode of nitrogenous elimination is related to the typical durations of emersion experienced by the fish. Intertidal species exposed on a regular cycle, and which may retain some contact with water, tend to remain ammoniotelic while reducing excretion rates in order to prevent excessive water loss. Amphibious fish that inhabit environments where emersion is less predictable than the intertidal, can store nitrogen during the state of emersion with some conversion to ureotelism or have been shown to tolerate high ammonia levels in the blood. Finally, the more amphibious fish are more adapted to moving on land and seeing in air. Structural modifications to the pectoral, pelvic, dorsal and anal fins, combined with a well‐developed musculature permit effective support and movement on land. For vision in air, there is a general trend for fish to possess close‐set, moveable, protruberant eyes set high on the head with various physical adaptations to the structure of the eye to allow for accurate vision in both air and water.     

急性离水操作胁迫对四指马鲅(Eleutheronema tetradactylum)幼鱼组织结构和氧化应激的影响

为探究急性离水操作胁迫对四指马鲅(Eleutheronema tetradactylum)幼鱼组织结构和氧化应激的影响，本研究通过组织切片和酶活性测定来观察及检测鳃和肌肉显微结构的变化、抗氧化酶活性以及Na+-K+-ATP酶活性，以探究四指马鲅应激反应规律。结果显示，急性离水操作胁迫2 h后，鳃小片、扁平上皮细胞以及线粒体丰富细胞都出现不同程度的损伤；肌肉从肌纤维变性、肌纤维束之间的间隙增宽、空泡化至肌肉组织整体失去固有形态，并且呈逐渐分解的趋势；超氧化物歧化酶(SOD)和还原型谷胱甘肽(GSH)有相同的变化趋势，胁迫后的2 h显著下降(P<0.05)，但12 h时则升至最高值。过氧化氢酶(CAT)和总抗氧化能力(T-AOC)水平在处理后2 h无显著变化(P>0.05)，之后升高，直到12 h达到峰值。丙二醛(MDA)含量在胁迫后的2 h无显著差异(P>0.05)，直到6 h达到峰值。SOD和CAT活性在处理24 h后显著低于处理前，其他酶变化不显著。Na+-K+-ATP酶活性在处理后2 h显著升高(P<0.05)，12 h达到峰值，24 h恢复到处理前水平。本研究表明，操作胁迫会对鳃和肌肉的组织结构造成损伤，而且随时间持续，损伤呈现加重趋势；肌肉抗氧化酶系统在受到氧化压力2 h后才启动，处理后12 h肌肉受到的氧化压力最大，抗氧化酶活性最强；在受到外界刺激时，SOD和GSH之间可能存在协同作用；MDA可以作为四指马鲅氧化损伤的快速响应生物标记物。