Morphology and distribution of granulomatous inflammation in freshwater ornamental fish infected with mycobacteria

Mycobacteriosis in fish is a chronic progressive ubiquitous disease caused by Mycobacterium marinum, M. gordonae and M. fortuitum in most cases. The aim of this study was to describe the morphology and distribution of lesions in 322 freshwater ornamental fish across 36 species. Granulomatous inflammation was diagnosed by gross examination and histopathology testing in 188 fish (58.4%); acid‐fast rods (AFR) were determined in only 96 (51.1%) fish from 19 species after Ziehl–Neelsen staining. The most often affected organs with AFR were the kidney (81.2%), digestive tract (54.1%), liver (48.2%), spleen (45.9%) and skin (21.2%); sporadically, AFR were found in the branchiae (9.4%) and gonads (4.7%). In 14 randomly selected fish originating from four different fish tanks, the distribution of mycobacterial infection was studied by culture examination of the skin, gills, muscle tissue, digestive tract, liver, spleen and kidney. In 12 fish, the species M. marinum, M. gordonae, M. fortuitum, M. triviale, and M. avium subsp. hominissuis (serotypes 6 and 8 and genotype IS901? and IS1245+) were detected; mixed infection caused by different mycobacterial species was documented in five of them.     

Detection and identification of aquatic mycobacteria in formalin-fixed, paraffin-embedded fish tissues

The isolation of mycobacteria from field samples is problematic, and isolation of the bacterium is sometimes not even attempted. The detection of mycobacteria through traditional histology using formalin‐fixed, paraffin‐embedded (FFPE) tissues is neither sensitive nor specific. However, detection of mycobacterial DNA from FFPE specimens, suspected of being infected with mammalian mycobacteriosis, is a routine clinical procedure. In the present study, a polymerase chain reaction (PCR)‐based method was used to detect and identify mycobacteria in FFPE specimens sampled from fish suspected of being infected with fish mycobacteriosis. A total of 45 fish tissue samples, comprising of 12 tissue samples obtained from experimentally infected fish and the remainder from fish naturally infected with mycobacteria, were analysed using a PCR protocol which amplifies a fragment of the mycobacterial 65 kDa heat‐shock protein (hsp65) gene. PCR‐restriction enzyme analysis and/or sequencing were employed to further analyse the PCR amplicons. The PCR results were compared with those obtained by histology and culture. Mycobacterial DNA was detected in 34 of the 45 samples examined, of which 16 samples (47%) showed granulomatous reactions on histological examination. Using histology as the gold standard, no false‐negative PCR results were obtained. Also, considering the presence or absence of granulomas as a diagnostic criterion, the sensitivity and specificity of PCR in 42 of the FFPE tissues were 16/16 (100%) and 8/26 (～30.8%), respectively. Corresponding microbiological cultures were available for 15 cases, of which 13 were pure Mycobacterium cultures. Of these, 13 were PCR positive (100% sensitivity and 50% specificity). The PCR‐based methods used here proved sensitive, specific and rapid for the detection of mycobacteria in routinely processed paraffin wax‐embedded and formalin‐fixed histological samples, and the results of the study suggest that this method has potential use in retrospective epidemiological studies.     

A low prevalence of mycobacteria in freshwater fish from water reservoirs, ponds and farms

A survey of the occurrence of mycobacteria was conducted from 717 freshwater fish (25 species) in two water reservoirs, five ponds and two farms in the Czech Republic. A total of 2182 tissue samples from these fish were examined using the conventional culture method. Thirteen mycobacterial isolates were obtained from 12 (1.7%) fish belonging to nine species. Isolates were identified using sequence analysis of the 16SrRNA gene as: Mycobacterium algericum, M. fortuitum, M. gordonae, M. insubricum, M. kumamotonense, M. nonchromogenicum, two isolates of M. peregrinum, M. terrae and M. triplex. Mycobacteria were isolated more frequently from fish skin and gills than from internal organs or muscles.     

Mycobacteria in aquarium fish: results of a 3‐year survey indicate caution required in handling pet‐shop fish

Fish are commonly infected with non‐tuberculous mycobacteria (NTM), which should be regarded as potential pathogens when handling aquarium fish and equipment. This study examined 107 aquarium fish from pet shops. Cultivation of the fish samples using different selective media was conducted for identification of NTM. Isolates were identified using the GenoType Mycobacterium common mycobacteria and additional species assays, sequencing of the 16S rRNA and rpoB genes, and real‐time PCR assay for identification of Mycobacterium (M.) marinum. Among the investigated fish, 79.4% (85/107) were positive for mycobacteria, with 8.2% (7 of 85) having two mycobacterial species present. Among the positive fish, the common pathogens M. marinum, Mycobacterium fortuitum (M. fortuitum group) and Mycobacterium chelonae were identified in approx. 90% of fish and other NTM species in 10%, including Mycobacterium peregrinum/septicum, Mycobacterium gordonae, Mycobacterium arupense, Mycobacterium kansasii, Mycobacterium ulcerans and Mycobacterium setense. The well‐known human pathogen M. marinum was present in 10.6% of the positive fish (9 of 85). The species of mycobacteria identified in the study are not only recognized as aquarium fish pathogens, but can also cause pathology in humans. Microbiological and clinical communities should therefore be sensitized to the role of NTM in infections associated with exposure to aquarium fish.     

大水面网围精养对水环境的影响及其对策

大水面网围精养对水环境的影响及其对策吴庆龙，陈开宁，高光，范成新，季江，隋桂荣，周万平（中国科学院南京地理与湖泊研究所，２１０００８）关键词网围养鱼，水环境，对策ＥＦＦＥＣＴＳＯＦＰＥＮＦＩＳＨＣＵＬＴＵＲＥＯＮＷＡＴＥＲＥＮＶＩＲＯＮＭＥＮＴＡＮＤ...     

A comparison of swimming capacity and energy use in seven European freshwater fish species

Abstract –  Migrating fish species with different swimming capacities and energy use show different capacities for passing obstacles between habitats, such as culverts and fish ladders. Here, we present an integrated study on swimming capacity and energetic use in seven European freshwater fish species with different ranges of migration (brown trout Salmo trutta L., European perch Perca fluviatilis L., roach Rutilus rutilus L., common carp Cyprinus carpio L., gudgeon Gobio gobio L., bullhead Cottus gobio L. and stone loach Barbatula barbatula L.). Critical ( U _crit), optimal ( U _opt) and maximum ( U _max) swimming speed and oxygen consumption ( M O₂) were analysed and showed values correlated to migration capacity with highest swimming capacities in trout and roach and lowest in stone loach and bullhead. The resulting data can be used to make estimates of maximum passable water speeds in culverts. In conclusion, long-distance migrators show higher swimming capacities and can potentially clear obstacles easier than short distance migrators with lower swimming capacities. Even small obstacles (<25 cm) could be a barrier for genetic exchange between populations in short-distance migrators.     

Immunohistochemical and Taqman real-time PCR detection of mycobacterial infections in fish

Real‐time PCR and immunohistochemistry (IHC) assays were developed to detect fish mycobacterial infections at the genus level, based on the RNA polymerase β subunit (rpoB) gene and polyclonal anti‐Mycobacterium rabbit serum, respectively. The PCR assay positively identified a number of pathogenic mycobacteria including Mycobacterium abscessus, M. avium ssp. avium, M. bohemicum, M. chelonae ssp. chelonae, M. farcinogenes, M. flavescens, M. fortuitum ssp. fortuitum, M. gastri, M. gordonae, M. immunogenicum, M. malmoense, M. marinum, M. montefiorense, M. phlei, M. phocaicum, M. pseudoshottsii, M. salmoniphilum, M. senegalense, M. shottsii, M. smegmatis, M. szulgi and M. wolinskyi. A detection limit equivalent to 10² cfu g^?1 was registered for M. salmoniphilum‐infected fish tissue. The IHC precisely localized both free and intracellular mycobacteria in tissues and detected mycobacterial infections down to 10² cfu g^?1 tissue. Both assays were found to be more sensitive than Ziehl–Neelsen (ZN) staining, where the detection limit was below 8 × 10³ cfu g^?1 tissue. Although specificity testing of the real‐time PCR against a panel of non‐Mycobacterium spp. revealed a degree of cross‐reaction against pure DNA extracted from Nocardia seriolae and Rhodococcus erythropolis, no cross‐reactions were identified (by either real‐time PCR or IHC) on testing of formalin‐fixed paraffin‐embedded (FFPE) tissues confirmed to be infected with these bacteria. The broad applicability of both assays was confirmed by analysis of FFPE tissues from a range of fish species infected with diverse Mycobacterium spp. The results indicate that both assays, alone or in combination, constitute sensitive tools for initial, rapid diagnosis of mycobacteriosis in fish. This should in turn allow rapid application of more specific studies, i.e. culture based, to identify the specific Mycobacterium sp. involved.     

玉曲河流域裂腹鱼类游泳能力研究及其在过鱼设施设计中的应用

为探究玉曲河流域裂腹鱼类的游泳能力,给过鱼设施建设和鱼类游泳行为学提供基础参数,本研究以该流域优势物种怒江裂腹鱼(Schizothorax nukiangensis)、贡山裂腹鱼(S. gongshanensis)为研究对象,在野外现场采用游泳能力测试水槽对裂腹鱼的感应流速、临界游泳速度、突进游泳速度及持续游泳速度、耐久游泳速度进行测定。测试结果表明：怒江裂腹鱼和贡山裂腹鱼的绝对感应流速相近,分别为0.18±0.01 m/s、0.18±0.06 m/s。怒江裂腹鱼和贡山裂腹鱼的绝对临界游泳速度分别为1.05±0.12 m/s、1.11±0.14 m/s,绝对突进游泳速度分别为1.45±0.26 m/s、1.40±0.21 m/s。二者的相对临界游泳速度分别为8.47±0.95 BL/s、9.21±2.45 BL/s,相对突进游泳速度均突破了十倍体长,分别为11.90±2.31 BL/s、10.80±3.16 BL/s。测得的怒江裂腹鱼和贡山裂腹鱼最大持续游泳速度均为0.63 m/s,怒江裂腹鱼和贡山裂腹鱼最大耐久游泳速度分别为0.93 m/s、0.73 m/s。当以怒江裂腹鱼、贡山裂腹鱼为主要过鱼对象时,建议鱼道入口及竖缝处流速1.05～1.40 m/s,过鱼设施内部的整体平均流速设计范围为0.24～1.05 m/s,以及休息池主流流速为0.18～1.05 m/s。本研究结果可为西藏玉曲河流域过鱼设施的设计提供参考,对该流域的鱼类资源保护具有重要意义。     

浅析我国观赏鱼产业发展及鱼种变化(上)

目前我国水产业以养殖为主,捕捞为辅。在2022年的水产品产量中,淡水养殖占47.91%,海水养殖占33.15%,捕捞占18.94%。在水产养殖中,鱼类养殖产量占52.16%,呈现出举足轻重的地位。“良种良养良销”是水产业中三个重要的组成部分,“良种”是水产业的龙头,“良养”是水产业的保障,加工、销售等“良销”环节是拉动水产业发展的重要动力。20世纪50年代以来,我国水产业取得了长足发展,为广大人民提供了丰富的水产品,解决了“吃鱼难”的问题。然而,目前我国水产业中有种无业、有业无种等现象还比较严重,没有从根本上解决“吃好鱼”及“吃放心鱼”的问题,为了加快我国水产业的发展,作者认为很有必要建立和实施“良种良养良销”体系,以推动我国水产业高质量发展。

     

Common trends in recruitment dynamics of north‐east Atlantic fish stocks and their links to environment,ecology and management

Recruitment dynamics are challenging to assess or predict because of the many underlying drivers that vary in their relevance over time and space. Stock size, demographic and trait composition, condition and distribution of spawning fish and the spatio‐temporal dynamics of trophic and environmental interactions all influence recruitment processes. Exploring common patterns among stocks and linking them to potential drivers may therefore provide insights into key mechanisms of recruitment dynamics. Here, we analysed stock‐recruitment data of 64 stocks from the north‐east Atlantic Ocean for common trends in variation and synchrony among stocks using correlation, cluster and dynamic factor analyses. We tested common trends in recruitment success for relationships with large‐scale environmental processes as well as stock state indicators, and we explored links between recruitment success and demographic, environmental and ecological variables for a subset of individual stocks. The results revealed few statistically significant correlations between stocks but showed that underlying common trends in recruitment success are linked to environmental indices and management indicators. Statistical analyses confirmed previously suggested relationships of environmental–ecological factors such as the subpolar gyre and Norwegian coastal current with specific stocks, and indicated a large relevance of spawning stock biomass and demographics, as well as predation, whereas other suggested relationships were not supported by the data. Our study shows that despite persistent challenges in determining drivers of recruitment due to poor data quality and unclear mechanisms, combining different data analysis techniques can improve our understanding of recruitment dynamics in fish stocks.     

Submerged light increases swimming depth and reduces fish density of Atlantic salmon Salmo salar L. in production cages

Artificial photoperiods that postpone sexual maturation and increase growth are now widely used in the Atlantic salmon Salmo salar L. farming industry. Few studies have been carried out to examine the effect of this treatment on fish behaviour and welfare in production cages. In this study, echo‐integration was used to observe the swimming depth and fish density of salmon in 20‐m‐deep production cages illuminated by lamps mounted above the water surface (SURF) or submerged in the cage (SUBS). From January to May, SUBS swam at a greater depth (5–11 m) than SURF (1–3 m) at night. SURF descended and SUBS ascended at dawn, but SUBS were still swimming at greater depth than SURF during the day from January to March. The difference in swimming depth resulted in SURF swimming at a median fish density about twice as high as SUBS at night and up to five times the calculated fish density. SURF increased the utilization of the cage volume as the biomass increased, but fish swimming at the highest density did so at up to 20 times the calculated fish density. The results suggest that salmon position themselves in relation to the artificial light gradient to maintain schooling behaviour and that the use of submersible lights may be a precaution to secure the welfare of caged salmon.     

Detection and molecular characterization of infectious spleen and kidney necrosis virus from major ornamental fish breeding states in Peninsular Malaysia

‘Gold standard’ OIE reference PCR assay was utilized to detect the presence of infectious spleen and kidney necrosis virus (ISKNV) in freshwater ornamental fish from Malaysia. From total of 210 ornamental fish samples representing 14 species, ISKNV was detected in 36 samples representing 5 fish species. All positive cases did not show any clinical signs of ISKNV. Three restriction enzymes analyses showed that the fish were infected by identical strains of the same virus species within Megalocytivirus genus. Major capsid protein (MCP) genes of 10 ISKNV strains were sequenced and compared with 9 other reference nucleotide sequences acquired from GenBank. Sequence analysis of MCP gene showed that all strains detected in this study were closely related to the reference ISKNV with nucleotide sequence identity that was ranging from 99.8% to 100%. In addition, phylogenetic analysis of MCP gene revealed that viruses from genus Megalocytivirus can be divided into three genotypes: genotype 1 include reference ISKNV and all other strains that were detected in this study, genotype 2 include viruses closely related to red sea bream iridovirus (RSIV), and genotype 3 include viruses closely related turbot reddish body iridovirus (TRBIV).     

Growth and morphology of an endangered native freshwater fish,crucian carp Carassius carassius,in an English ornamental pond

• 1. Crucian carp Carassius carassius, which is native to south‐east England, has received little previous study in Britain but is threatened by introductions of goldfish Carassius auratus through hybridization and by frequent mis‐identification of brown goldfish as crucian carp. The present study provides the first data on back‐calculated growth, morphology and age‐at‐maturity of crucian carp in Britain.
• 2. The youngest mature crucian were female (age 2+), the smallest mature crucian were male (age 3+), with almost all fish mature at age 3+. Sex dimorphism in back‐calculated standard length at age was not observed except at age 2+ (juveniles longest). Shifts in morphological characters often associated with maturation (dorsal and ventral fin length, dorsal and anal fin depth, pre‐dorsal distance, body width) were observed between ages 4+ and 7+, which (in contrast to the relatively abrupt shift in maturity) suggests that age and size at maturation may not reflect the size and age of active (i.e. real) spawning.
• 3. Back‐calculated standard lengths at age for crucian carp were intermediate relative to other European populations, indicating that environmental conditions in England appear to be well suited to the species despite being at the westernmost extent of the species' native European range. The management of ornamental ponds specifically for crucian carp is discussed.

© Crown copyright 2007. Reproduced with the permission of Her Majesty's Stationery Office. Published by John Wiley & Sons, Ltd.     

Effects of within-patch habitat structure and variation on fish assemblage characteristics in the Bernecei stream, Hungary

Abstract –  Over a 3-year period we examined variability in physical habitat structure and species richness, abundance and assemblage composition of fishes in 13 habitat patches in the Bernecei stream, Hungary. Principal component analysis of habitat structure data from patches elucidated a riffle-run-pool habitat gradient across patches. Temporal habitat variability increased significantly from riffle to pool patches. Fish assemblage characteristics displayed relatively continuous change over the habitat gradient and were relatively stable within patches. Assemblage structure properties (e.g., species richness) displayed different responses to the habitat gradient and to within-patch habitat variability. In general, pool patches had more diverse assemblages and greater within-patch assemblage variability than riffle patches. However, within-patch dynamics were largely determined by the population dynamics of a habitat generalist (i.e., minnow). Broad scale environmental variability (i.e., a catastrophic 100-year flood) also appeared to affect within-patch fish assemblage characteristics. Our results demonstrate that fish assemblage structure is influenced by physical variability (i.e., both floods and spatio-temporal habitat variability) within the Bernecei stream.     

硝化细菌对淡水水族箱水质及养殖观赏鱼影响的初步研究

本试验设立硝化细菌、水生观赏植物对照组与空白对照组进行对比试验。结果表明硝化细菌对降低淡水水族箱中COD、NH4^ -N、NO^2 -N效果显。有利于提高蓝毛足鲈等的相对生长率，对孔雀鱼、七彩神仙鱼增色有益。实验中还发现硝化细菌有明显预防细菌性肠炎病发生的作用。硝化细菌在淡水水族箱中固着种群稳定需要25—28天，在施用硝化细菌3—4周后水体COD、NH4^ -N、NO^2 -N达到稳定水平。     

Dominant pathogenic species of mesophilic aeromonads isolated from diseased and healthy fish cultured in Poland

Aeromonas isolates were collected from cultured fish, characterized phenotypically and identified to species using 16S rDNA. The pathogenicity of all isolates was assayed on the basis of haemolytic and proteolytic activity and challenge tests were performed for isolates from healthy fish. A total of 131 Aeromonas isolates were obtained and identified as follows: A. hydrophila (13), A. bestiarum (23), A. salmonicida (motile biogroup) (19), A. caviae (2), A. sobria (18), A. veronii bt. sobria (42), A. jandaei (1), A. encheleia (11) and A. allosaccharophila (2). All isolates of A. hydrophila and A. bestiarum and most isolates of A. salmonicida and A. veronii were classified as pathogenic. Aeromonas hydrophila was isolated only from diseased trout except for one isolate obtained from carp fry. The other potentially pathogenic Aeromonas species were present in diseased as well as healthy fish. The pathogenicity of isolates from healthy fish was correlated with their enzymatic activity and was also tested by challenge experiments. The dominant pathogenic species were A. veronii bt. sobria, A. bestiarum and A. salmonicida in common carp and A. hydrophila in rainbow trout.     

Distribution and properties of lactate dehydrogenase isoenzymes in red and white muscle of freshwater fish

The distribution and kinetics of LDH isoenzymes in red and white muscles of 5 species of salmonids, 4 species of cyprinids and one coregonid species were studied. In all species the white muscles are characterized by the occurrence of only the most cathodic isoenzymes, or groups of isoenzymes. The red muscles contained either the full set of isoenzymes (cyprinids) or a selection in which the anodic forms dominated (salmonids, coregonid). The most striking difference between the two types of muscle was met inCoregonus sp. The temperature profiles of pyruvate affinity are similar in all species of fish studied. On the other hand, K_m(pyr) values and degree of pyruvate inhibition are closely related and vary greatly with temperature, with the taxonomic position (and thus biology) of the species, and with electrophoresic mobility of the isoenzyme. Highest affinity and strongest inhibition occurred in the anodic (H₄) isoenzymes of cyprinids at low temperature; lowest affinity and zero inhibition in the cathodic isoenzymes (M₄ M₄) of salmonids and coregonids at high temperature. In salmonids the more recently duplicated loci of the M-group of isoenzymes possess identical K_m values at all temperatures, whereas the two older M and H loci differ greatly in this respect. Thus the more recent duplication of LDH loci in salmonids and coregonids may be seen as a mechanism by which the tetramers required for LDH activity can be constructed from more closely related subunits than are provided by the older M and H loci.Some problems in connection with the determination of the kinetic constants of the lactate oxidase reaction are discussed and it is suggested that an alkaline, pyruvate trapping system provides conditions which are more realistic than those of other assay systems. The K_m(lactate) values found are in the biological range and, at 20°C, provide further circumstantial evidence that the red muscles of fish should be capable of oxidizing the lactate produced by the white muscles during strenuous exercise. At 4°C the K_m(lactate) values are abnormally high in all muscle preparations and thus are not correlated with the K_m(pyruvate) values which are lowest at this temperature.     

渤海鱼类群落功能群及其主要种类

根据2009年8月和10月对渤海渔业资源的底拖网调查,采用胃含物分析、聚类分析和功能群划分的方法研究了渤海鱼类群落的功能群组成及其主要种类。结果表明,渤海夏、秋季鱼类群落包括7个功能群,分别为浮游动物食性功能群、杂食性功能群、底栖动物食性功能群、虾食性功能群、虾/鱼食性功能群、鱼食性功能群和广食性功能群;其中主要功能群为浮游动物食性功能群、杂食性功能群和虾/鱼食性功能群;主要种类有小黄鱼、蓝点马鲛、斑、赤鼻棱鳀、银鲳和黄鲫。圆筛藻、中华哲水蚤、太平洋磷虾、长额刺糠虾、中国毛虾、甲壳类幼体、日本鼓虾、六丝矛尾虎鱼、小黄鱼、双壳类和腹足类是当前渤海夏、秋季鱼类群落的主要饵料种类。     

Distribution of infectious pancreatic necrosis virus (IPNV) in wild marine fish from Scottish waters with respect to clinically infected aquaculture sites producing Atlantic salmon, Salmo salar L

This study represents the first large-scale investigation of IPNV in Scottish wild marine fish. Kidney samples were taken from 30 627 fish comprising 37 species and 45 isolations were made from nine different species, illustrating these as reservoirs of IPNV in Scottish waters. The estimated prevalence of IPNV in the Scottish marine environment was low at 0.15% (90% confidence intervals, (CI) of 0.11-0.19%). This was significantly greater in fish caught less than 5.0 km from IPN-positive fish farms in Shetland, at 0.58% (90% CI of 0.45-0.77%). This prevalence persisted and did not significantly decrease over the 16-month period of study. The estimated prevalence of IPNV for each positive species was less than 1% with the statistically non-significant exceptions of flounder, Platichthys flesus (L.), at 12.5% (90% CI of 0.64-47.06%) and saithe, Pollachius virens (L.), at 1.11% (90% CI of 0.49-2.19%). The 45 isolates were titrated and all but two were below the detection limit of the test (<55 PFU g(-1)). Titres of 3.8 x 10(2) PFU g(-1) and 2.8 x 10(1) PFU g(-1) were calculated from common dab, Limanda limanda (L.), and saithe, respectively. This study provides evidence that clinical outbreaks of IPN in farmed Atlantic salmon may cause a localized small increase in the prevalence of IPNV in wild marine fish.