Three‐dimensional visualization of green fluorescence protein‐labelled Edwardsiella tarda in whole Medaka larvae

The invasive fish pathogen Edwardsiella tarda is common in aquatic environments and causes the environmentally and economically destructive emphysematous putrefactive disease called edwardsiellosis. In order to understand the organism's infection pathway, medaka larvae (Oryzias latipes) were immersion‐infected with E. tarda labelled with green fluorescence protein (GFP) and then visualized in three dimensions under confocal laser microscopy and light‐sheet fluorescence microscopy. Confocal microscopy revealed GFP‐labelled E. tarda in the mouth, head, gill bridges, gill cover, skin, membrane fin, gastrointestinal tract and air bladder, and in the caudal vein, somite veins, caudal artery and caudal capillaries. Light‐sheet microscopy additionally showed GFP‐labelled E. tarda in the pharyngeal cavity, muscle of the pectoral fin and cardiac atrium and ventricle. These findings suggest that during its infection of fish, E. tarda initially adheres to, and invades, the epithelial cells of the skin, gills and gastrointestinal tract (through the pharyngeal cavity); E. tarda then enters the blood vessels to access organs, including the air bladder and heart.     

WSSV envelope protein VP51B links structural protein complexes and may mediate virus infection

White spot syndrome virus (WSSV), an enveloped double‐stranded DNA virus, is the causative agent of a disease that has led to severe mortalities of cultured shrimps in Taiwan and many other countries. In the previous study, Penaeus monodon chitin‐binding protein (CBP) and glucose transporter 1 (Glut1), two cell membrane proteins, were found to at least interact with other 10 WSSV envelope proteins including VP51B. These envelope proteins might form a protein complex. According to the known information, VP51B was used to identify its role in the protein complex. Western blotting of the intact viral particles and fractionation of the viral components confirmed that VP51B is one of WSSV envelope proteins. In this study, the protein–protein interaction between VP51B and other WSSV envelope proteins was identified by far‐western blot experiment and VP51B was found to interact with VP24, VP31, VP32, VP39B and VP41A. Furthermore, the in vivo neutralization experiment using recombinant VP51B plus with VP39B showed the best inhibition. These data indicate that VP51B participates in the WSSV protein complex and plays an important role in WSSV infection.     

Mycobacteriosis caused by Mycobacterium marinum in reared mullets: first evidence from Sardinia (Italy)

Mycobacterium marinum is a slow‐growing non‐tuberculous mycobacterium, and it is considered the most common aetiologic agent of mycobacteriosis in wild and cultured fish. The diagnosis is principally made by histology when positive Ziehl–Neelsen stain granulomas are detected. The aim of this study was to investigate the occurrence of mycobacteriosis in extensively cultured Mugilidae of two lagoons (Cabras and San Teodoro) from Sardinia by the use of histology, microbiology, PCR and DNA sequencing. Nine of 106 mullets examined were affected by mycobacteriosis, and the spleen was the most affected organ. The histology detected higher rate (100%) of infection in spleen than the culture and PCR (75% and 62.5%, respectively). The sequencing of hsp65 gene identified M. marinum as the primary cause of mycobacteriosis in the mullets examined. Mullets affected by mycobacteriosis were mainly fished in the San Teodoro lagoon characterized by critical environmental conditions. Histology remains the most common method in detecting fish affected by mycobacteriosis, and PCR‐based methods are essential for species identification. Our finding are worthy of attention because mycobacteriosis caused by M. marinum in reared mullets was evidenced for the first time in Sardinia, suggesting that this disease may be underestimated also in other cultured fish species.     

Pansteatitis in polluted Olifants River impoundments: nutritional perspectives on fish in a eutrophic lake,Lake Loskop,South Africa

This study compares the aetiology of pansteatitis in Lake Loskop, relative to two other impoundments along the Olifants River. Macroscopic and microscopic pathology, age determination and analysis of stomach content, fatty acids and stable isotopes explain the high prevalence of pansteatitis in Oreochromis mossambicus (Peters) and several other species in Lake Loskop. All the dietary indicator comparisons between pansteatitis‐affected and healthy fish fail to support a systemic cause. Pansteatitis in Lake Loskop was linked to size and weight of O. mossambicus, but not to ontogenic age. Fish in Lake Loskop showed abnormally high omega‐3 to omega‐6 fatty acid ratios normally only found in marine fish with no significant difference in degree of assimilation of these fatty acids between pansteatitis‐affected and healthy fish. This explains the vulnerability to, but not the occurrence of, pansteatitis. As a cause for the pansteatitis, these results point towards sporadic vitamin E‐depleting trigger events, known sporadic fish die‐off occurrences that provide surviving fish with a rich source of rancid fats on which to scavenge. The mechanism ties pansteatitis to eutrophication and trophic cascade effects, the intrinsic drivers of the disease and suggests an adaptive management strategy that might be applied by relevant conservation authorities.     

Preservation causes shrinkage in seahorses: implications for biological studies and for managing sustainable trade with minimum size limits

• 1. The implications of shrinkage associated with desiccation and ethanol preservation for seahorses (genus Hippocampus) were investigated using Hippocampus guttulatus (European long‐snouted seahorse) as a model. Specifically, this research addressed the implications of preservation for taxonomy and life history studies and the application of minimum size limits (MSL) for managing seahorse trade.
• 2. In 2004, the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) listed all seahorse species on its Appendix II, and recommended a 10 cm MSL as an interim means of ensuring sustainable international trade. Inconsistencies in seahorse measurement methods and repeatability posed challenges for applying the MSL. Moreover, the shrinking effect of desiccation on body length observed in other fish was assumed to be negligible for seahorses because of their high degree of ossification.
• 3. Changes in seahorse sizes were measured following immersion in ethanol and desiccation. H. guttulatus shrank on average by 0.1–2.3% when preserved in ethanol and 3.0–6.4% when dried, depending on the trait measured. Similar trends were observed in a sample of H. kuda (yellow seahorse). Specimen posture during drying, and measurement methods also influenced estimates of size.
• 4. Based on the shrinkage observed, 14–44% of captured seahorses that are dried could shrink to below the recommended MSL, even if all seahorses were longer than the MSL at capture. This demonstrates that small changes in body lengths can have significant implications for trade of species managed with size limits.
• 5. Recommendations are to (1) standardize seahorse measurement methods, (2) consider the effects of preservation and measurement technique on body lengths, and apply appropriate corrections in comparative studies and when developing fisheries management strategies, and (3) adjust size limits at the point of capture to ensure retained seahorses comply with the CITES recommended MSL.

Copyright © 2008 John Wiley & Sons, Ltd.     

Status,ecology and conservation of an endemic fish,Oreochromis niloticus baringoensis,in Lake Baringo,Kenya

• 1. The tilapia Oreochromis niloticus baringoensis is a genetically and morphologically distinct sub‐species of Oreochromis niloticus endemic to Lake Baringo, Kenya. In recent years, concern has been expressed as to its status. Recent declines in catch returns suggest the population may be threatened, with conservation action required to safeguard it.
• 2. Catch returns from the Baringo fishery since 1964 have shown considerable fluctuations for all species, but especially O. n. baringoensis. From a peak of 712 t in 1970, their total catch was only 5 t in 2005, despite a 2 year period of closure in 2002 and 2003. Changes in fishery catch and relative abundance were independent of exploitation in the fishery but were significantly correlated with lake level.
• 3. Few individuals were captured at lengths >250 mm, with no fish sampled >284 mm. During periods of high lake level, individuals matured at smaller sizes and were capable of growing to larger ultimate sizes. With maturity at lengths <130 mm and fishery regulations preventing removal of fish <180 mm, there was a relatively large proportion of mature fish that was below exploitation size each year (19 to 44%). In most years, the proportion of fish available for exploitation was <10%.
• 4. Stable isotope analyses revealed O. n. baringoensis was reliant upon planktonic basal resources and zooplankton carbon. There was only minimal trophic overlap with other fish species in the lake, indicating little potential for competition for food resources.
• 5. These data suggest that the population status of O. n. baringoensis is not threatened per se, but subject to an unpredictable and unstable environment that strongly influences their life‐history traits and, ultimately, their population abundance, and should be managed accordingly.

Copyright © 2008 John Wiley & Sons, Ltd.     

Mixed stream channel morphologies: implications for fish community diversity

• 1. Stream classification systems are widely used in stream management and restoration. Whereas the principal morphological types of these classification systems are increasingly recognized for their ecological connections, the roles of intermediate and mixed morphologies are still poorly understood, yet may be biologically significant.
• 2. Twenty‐five stream reaches in north‐western Vermont were classified by channel morphology to determine whether fish community diversity differed among pool‐riffle, mixed (i.e. pool‐riffle/cascade, pool‐riffle/other) and forced pool‐riffle stream morphological groups. Stream reach surveys included cross‐sectional surveys, longitudinal profiles, bed substrate characterization, and fish surveys.
• 3. Three fish community diversity measures were calculated: (1) species richness (S); (2) Shannon–Weaver Index (H′); and (3) Simpson's Index (1/D). Multivariate analysis of covariance (MANCOVA) followed by analysis of variance (ANOVA) were used to explore potential differences in fish diversity among stream morphological groups. Fish diversity was significantly different for all three community diversity measures (P?0.05), with pool‐riffle/cascade morphology consistently exhibiting the greatest fish diversity and forced pool‐riffle the lowest.
• 4. These results suggest that fish community diversity is significantly associated with distinct channel morphologies. Generally, pool‐riffle/cascade and pool‐riffle/other stream morphological groups supported habitats that fostered greater species diversity than more homogeneous and uniform pool‐riffle reaches. The observed patterns of diversity are likely to be the result of habitat patches created by variations in flow and other physical characteristics in reaches of mixed morphologies.
• 5. These results support fish sampling schemes that incorporate morphological heterogeneity, such as proportional‐distance designation. Sampling strategies that focus on homogeneous reaches may underestimate diversity, and misrepresent stream condition when fish community data are used in indices of biological integrity (IBIs). Reaches of mixed stream morphologies should be recognized as areas of biological importance in stream and catchment management and in conservation efforts.

Copyright © 2008 John Wiley & Sons, Ltd.     

Habitat selection of freshwater‐dependent cetaceans and the potential effects of declining freshwater flows and sea‐level rise in waterways of the Sundarbans mangrove forest,Bangladesh

• 1. Generalized additive models of sighting data for cetaceans collected during two surveys of waterways in the Sundarbans mangrove forest of Bangladesh indicated that Ganges River dolphin Platanista gangetica gangetica distribution was conditionally dependent (P<0.05) on low salinity, high turbidity, and moderate depth during both low and high freshwater flow; and Irrawaddy dolphin Orcaella brevirostris distribution was conditionally dependent (P<0.05) on low salinity during high freshwater flow, high and moderate depths during low and high freshwater flow, respectively; low and high‐low extremes of turbidity during low and high freshwater flow, respectively; and high temperature and increasing numbers of large–small channel confluences during low freshwater flow.
• 2. According to sighting data collected over a 3‐year period by the captains of three nature tourism vessels, there were significant differences between the actual and expected frequencies of Ganges River dolphin sightings and individuals according to various channel types (chi‐square=64.22, P<0.0001 and chi‐square=134.14, P<0.0001, respectively, df=6) and of Irrawaddy dolphin sightings and individuals (chi‐square=15.28, P=0.0182, and chi‐square=29.42, P<0.0001, respectively, df=6), with shared preferences for wide sinuous channels with at least two small confluences or one large confluence.
• 3. The dependency exhibited by both species for environmental characteristics associated with abundant freshwater flow, including low salinity and the availability of confluences, make them particularly vulnerable to habitat loss due to upstream water abstraction and sea‐level rise.
• 4. Although the results of this study may not affect plans for construction in India of large‐scale, inter‐basin water transfer projects that will result in further declines in freshwater flows, or decisions within the international community about CO₂ emissions affecting global sea levels, they can be used to prioritize locations where protective measures could be employed to benefit the long‐term conservation of both species.

Copyright © 2008 John Wiley & Sons, Ltd.     

Geosmin depuration from European eel (Anguilla anguilla) is not affected by the water renewal rate of depuration tanks

This study established that geosmin depuration from European eel is not affected by the water renewal rate of depuration tanks. A general fish bioaccumulation model extended with terms that account for effects of tank water renewal rate and system losses of chemicals, predicted strong effects of the water renewal rate of depuration tanks on geosmin depuration from European eel. Model predictions were validated in a depuration experiment with geosmin‐loaded European eel (n = 95) with a mean (SD) individual weight of 134.4 (5.0) g and a mean (SD) lipid content of 33.7 (2.8) % (w/w). Fish were depurated for 24, 48 or 72 h at three different tank water renewal rates (0.3, 3.3 and 33 day^?1). Treatments were installed by three different mean (SD) water flow rates (13.8 (1.3), 143.5 (9.2) and 1511 (80) L kg fish^?1 day^?1) over 30‐L tanks. Eels eliminated geosmin from their bodies, but unlike the model predicted, this was independent of the water renewal rate of the depuration tanks. Although being eliminated from the fish, geosmin hardly appeared and certainly did not accumulate in the water of the depuration tanks as the model predicted. This observation may be explained by geosmin being eliminated from eel as metabolite rather than the parent compound. Geosmin elimination from eel seems not to occur according to the generally accepted passive diffusion mechanism for excretion of lipophilic chemicals, and geosmin biotransformation by the eel seems indicated. Clearly geosmin depuration from European eel cannot be enhanced by increasing water renewal rates of depuration tanks.