Outbreak of mass mortality of yearling groupers of Epinephelus (Perciformes,Serranidae) associated with the infection of a suspected new enteric Sphaerospora (Myxozoa: Myxosporea) species in South China Sea

A suspected new enteric Sphaerospora species was believed to be directly associated with the mass mortality of yearling groupers of Epinephelus spp. in South China. The epizootic generally emerged from late September to late April of the following year. The infection prevalence and mortality rate were significantly negatively correlated with fish size. Clinical signs included anorexia, cachexia and extrusion of white pulp‐like substance from anus after gentle pressure on the abdomen. Upon necropsy, severe intestinal oedema, thin and transparent intestinal wall, swollen spleen, kidney and gall bladder could be observed. Wet preparation of the infected samples showed large amount of typical disporous plasmodia of the genus Sphaerospora, but no mature spores were observed. Epidemiological investigation showed that this parasite exclusively infected Epinephelus groupers. Histopathologically, this species mainly infected the epithelium of intestine and kidney tubules and caused severe epithelia sloughing and the collapse of intestinal villus. Interestingly, this enteric myxosporidiosis did not cause severe emaciation of infected fish for mass mortality usually emerged within 2–3 days after appearance of clinical signs. The species was most genetically related to Sphaerospora fugu (89% sequence identity) and phylogenetically positioned within marine Sphaerospora lineage. This is the first report of enteric sphaerosporosis of groupers.     

Abnormal mortalities of the carpet shell clam Ruditapes decussatus (Linnaeus 1756) in natural bed populations: a practical approach

In situ controlled systems based on PVC cages surrounded by a net mesh and buried in sand were designed to determine the causes of important mortalities in producing locations of carpet shell clam, Ruditapes decussatus, in two important production areas of Galicia (NW Spain). The pathogenic status of the clam populations (focusing mainly on Perkinsus sp. and clam haplosporidian) as well as changes in water temperature and salinity were recorded. Sampling was conducted during a short period of time when high mortalities were present in the two locations and in a 1‐year study when no important mortalities were registered. We found that the environmental conditions and not the presence of pathogens seemed to be related to mortalities in the study. Our results suggest that mortality rates, especially in natural beds, can emerge from a complex synergy of different factors. Moreover, we propose in situ controlled systems based on PVC cages as a useful method to control infaunal bivalve mortalities in natural beds.     

Behaviour,escapement and mortality of female European silver eels within a regulated lowland river draining into the Baltic Sea

Migration patterns of 146 female silver eels on their way through fresh and brackish water stretches of the Warnow River into the Baltic Sea were studied using acoustic telemetry. The majority of eels (77%) regained their expected downstream migration behaviour after a median rehabilitation time of 2 days. Positive rheotactic behaviour was observed for 23% of tagged eels after release, probably influenced by experimental procedures. The observed silver eel escapement rate was 63%. Two migratory groups were classified by significantly different migratory speed, time of residence in brackish waters and biometric silvering characteristics. Although the classification of silvering stages is based on these measures, the proportions of these stages did not reflect the observed differences in migratory behaviour. In the context of a permanent silver eel descent in the freshwater part of the Warnow River, a permanent escapement was likewise occurring into the Baltic Sea.     

Detection and characterization of a rhabdovirus causing mortality in black bullhead catfish,Ameiurus melas

This study fully describes a severe disease outbreak occurred in 2016 in black bullhead catfish farmed in Italy. Affected fish showed nervous clinical signs as well as emaciations and haemorrhagic petechiae on the skin at the fin bases, abdomen and gills. Viral isolation in cell culture allowed the subsequent identification of a rhabdovirus, tentatively named ictalurid rhabdovirus (IcRV), through electron microscopy, immunofluorescence and whole genome sequencing (WGS). The newly isolated virus, together with 14 additional viral strains stored in our repository and detected during similar mortality episodes in the period 1993–2016, was phylogenetically analysed on the basis of the nucleoprotein and the glycoprotein nucleotide and amino acid sequences. The genetic distances among Italian IcRV strains were also estimated. Our results show that all the IcRV strains belong to the genus Sprivivirus and are closely related to the tench rhabdovirus (TenRV). Italian catfish production is constantly decreasing, mainly due to viral infections, which include the newly characterized IcRV. Data presented in this work will assist to investigate the molecular epidemiology and the diffusive dynamics of this virus and to develop adequate surveillance activities.     

Lethal effect of filamentous algal blooms on Atlantic herring (Clupea harengus) eggs in the Baltic Sea

1. During this study, the effects of epiphytic filamentous algae on the survival of demersal spawned fish eggs were investigated in one of the most important spawning grounds of herring in the western Baltic Sea, which is subject to intense and ongoing eutrophication.
2. In coastal marine ecosystems all over the world, eutrophication is a primary water quality issue, often resulting in mass developments of bloom‐forming algae. Macro‐algal blooms have immense ecological effects, as they alter the structure and the function of an ecosystem. Numerous fish species are affected, as they depend on those coastal areas for spawning and as juvenile habitats.
3. A comparison of the survival of herring eggs on two natural spawning beds re‐vealed an immense impact of a filamentous algal bloom. The mortality rate reached nearly 100% in the area with massive occurrence of filamentous brown algae.
4. Hypothesizing that the presence of filamentous algae facilitates herring egg mortality, field and laboratory experiments were conducted and revealed particular effects of distinct algal species. The survival rate of artificially spawned eggs on algal substrates and control substrates was compared. Whereas filamentous forms of the green alga Ulva intestinalis induced no immediate effect, significantly higher egg mortality was documented in experiments with the filamentous brown alga Pylaiella littoralis.
5. Considering the ecological and economical importance of herring and other coastal spawners on the one hand and the persisting and increasing effects of eutrophication and climate change on the other hand, the results of our study clearly underpin the necessity to increase global efforts to reduce nutrient loads in coastal waters.

     

Nematode parasites infecting the starry batfish Halieutaea stellata (Vahl) (Lophiiformes: Ogcocephalidae) from the East and South China Sea

The starry batfish Halieutaea stellata (Vahl) is a small, benthic fish found in Indo‐West Pacific Oceans. However, our present knowledge of the helminth parasites of this fish is still fragmentary. In this study, a total of 29 fish collected from the East and South China Sea were examined to determine the prevalence, intensity and species composition of helminth parasites in H. stellata. Using morphological and molecular approaches, four species of nematodes were found parasitic in this fish host, including the adults and fourth‐stage larvae of Raphidascaroides nipponensis Yamaguti 1941; adults and third‐stage larvae of Raphidascaris lophii (Wu 1949), third‐ and fourth‐stage larvae of Hysterothylacium larval type IV‐A of Shamsi, Gasser & Beveridge 2013 and third‐stage larvae of Hysterothylacium amoyense (Hsü 1993). Halieutaea stellata represents a new host record for the three last‐named nematodes. Raphidascaroides nipponensis with the highest prevalence (82.5%) and intensity (mean = 13.5) of infection was considered as the dominant parasite species in H. stellata. The detailed morphology of the different developmental stages of the four nematode species was studied using light and scanning electron microscopy. All nematode species were also genetically characterized by sequencing and analysing the internal transcribed spacer (ITS) of the ribosomal DNA. This study provides further data on the occurrence of nematode parasites in H. stellata and also contributes to facilitate an accurate and rapid diagnosis of the infection by these little‐known nematodes.     

Observation and quantification of two incidents of mass fish kill of Icelandic summer spawning herring (Clupea harengus) in the winter 2012/2013

The Icelandic summer‐spawning herring (Clupea harengus) stock overwinters in large, dense schools like other herring stocks. In the winter of 2012/2013 around 300 thousand tonnes, or ~70% of the spawning stock, overwintered in a fjord west of Iceland. The inner part of the fjord, where the herring was located, is separated from the outer part with a natural barrier and a built‐up road with a 210 m long bridge. This creates strong tidal currents under the bridge. On 14 December and again on 1 February mass mortalities of herring took place in this location, and the sea floor and the shores were covered with dead herring. Fieldwork, including camera and video recordings on the shore and on a small boat, was conducted 3 and 4 days after the incidents. Results from this indicated that a total of 55 thousand tonnes had died during these two incidents, an amount nearly equal to the total catch from the stock that year. Measurements of environmental conditions in the days following the incidents showed that the oxygen saturation was generally 20%–40% but was as low as 15% (1.1 ml/L). The most likely explanation for the mortalities was oxygen depletion resulting from respiration of the large herring biomass, limited atmospheric‐water gas exchange due to calm and cold weather prior to both incidents and sea ice on part of the fjord, and limited renewal of water coming in and out via tidal currents. Aerobic decomposition of dead herring came additionally in the latter incident.     

Impaired larval development in broods of eelpout (<Emphasis Type="Italic">Zoarces viviparus</Emphasis>) in Danish coastal waters

Eelpouts (Zoarces viviparus), a viviparous fish, were sampled in Danish coastal waters during October and November 2001 and 2002, in ten different areas, which are receiving effluents from cities and industry to more or minor degree. The presence of gross abnormalities in eelpout broods has been suggested to be a useful biomarker of the impact of hazardous substances on fish reproduction in the marine environment as chronic exposure to various substances has the potential to induce severe developmental defects in fish embryos and larvae. Relatively high frequencies of female eelpouts (20–53) with elevated levels (>5) of larvae with developmental defects in the broods were found in four shallow fjords with effluents from larger cities and industry compared to areas. Deformations like spiral or bend shapes of the spinal axis, cranio-facial defects, eye lesions or loss of eyes were the dominating types. In some ofthe areas with the highest incidences of developmental defects, the adults had significantly enlarged livers, which may also be a possible effect caused by contaminant exposure. In two of the areas, relatively high frequencies of the broods contained larvae, which had died late but without any visible developmental defects. These two areas were affected by severe oxygen depletion prior to sampling time suggesting that examination of broods in the eelpout may include not only impact of hazardous substances but also effect of eutrophication-related problems on fish reproduction in the marine environment.     

An investigation on the gamete quality of Black Sea trout (Salmo trutta labrax) broodstock fed with mealworm (Tenebrio molitor)

In this study, the effect of combined feeding Black Sea trout (Salmo trutta labrax) broodstock with commercial feed and mealworm larvae (Tenebrio molitor) for 50 days before the breeding period in order to evaluate the gamete quality. While the control group was fed with commercial feed, mealworm larvae were given as an additional protein source 2 days a week and 3 days a week to the experimental groups formed from female and male individuals. In addition to the growth parameters, the number of eggs (number/individual) and the egg diameter (mm), sperm volume (ml), density of spermatozoa (×10⁹ cell/ml), total motility (%), progressive motility (%) and average curvilinear velocity (VCL, µm/sec) values were determined. At the end of the study, the highest live weight gain was found as 46.2 g in the control group of male. The egg diameters were 4.3 ± 1.8 mm and 4.5 ± 1.4 mm in the worm treatment groups fed twice and three times with mealworm respectively. As a result, it was determined that feeding fish with mealworm larvae as an addition to the commercial diet in female individuals did not affect the amount of eggs (p > .05), but the egg diameters were significantly smaller in the control group than the mealworm groups (p < .05). In spermatological characteristics, only the amount of sperm was different between the groups (p < .05); all other parameters were found to be similar to each other with no statistical differences (p > .05).     

Morphological and genetic characterization of Hysterothylacium Ward & Magath, 1917 (Nematoda: Raphidascarididae) larvae in horse mackerel,blue whiting and anchovy from Spanish Atlantic and Mediterranean waters

The presence of zoonotic Hysterothylacium larvae in fish from Spanish Atlantic and Mediterranean waters, which can cause economic losses for commercial fisheries, has been reported in several studies; however, little is known about species identity in this region. The aim of this study was to identify at species level the Hysterothylacium morphotypes detected in three commonly consumed fish: horse mackerel (Trachurus trachurus), blue whiting (Micromesistius poutassou) and anchovy (Engraulis encrasicolus). Third‐ and fourth‐stage Hysterothylacium larvae, as well as adults obtained from larval in vitro culture, were morphologically and molecularly identified by ITS1/ITS2 rDNA sequencing. Four Hysterothylacium morphotypes were detected. Genetic analysis showed that morphotypes VIII and IX were different larval stages of Hysterothylacium aduncum, which was supported by cultured adult species identification. Morphotypes III and IV were found to correspond to different developmental stages of another species of Hysterothylacium. As all larval types detected were morphologically indistinguishable from others previously reported yet showed clear genetic differences, they are referred here as new genotypes. This is the first time that ITS‐sequence data of various developmental stages of the same species, including adults, have been studied and compared, providing crucial knowledge for future studies on Hysterothylacium identification and biology.     

Evidence of fish and human pathogens associated with doctor fish (Garra rufa,Heckel, 1843) used for cosmetic treatment

Doctor fish (Garra rufa, Heckel, 1843) are increasingly used for cosmetic treatment raising particular concerns regarding the potential transmission of infections to clients. Investigations of microbial causes undertaken in two outbreaks of mortality among G. rufa used for cosmetic treatment revealed the presence of multiple bacteria, including both fish and human pathogens such as Aeromonas veronii, A. hydrophila, Vibrio cholerae, Shewanella putrefaciens, Mycobacterium marinum and M. goodii. This range of bacteria indicates an intense microbial proliferation involving multiple pathogens, most likely induced by the poor health condition of the fish. Most of the detected pathogens are well‐known agents of zoonosis. Indeed, M. goodii is an emerging nosocomial human pathogen that has never been detected in fish to date, nor in other animals. This first detection of M. goodii associated with fish infection points out a new zoonotic potential for this pathogen. These findings point out that handling, poor environmental conditions and the presence of fish pathogens, that can compromise the immune system of fish, can result in a mixed microbial proliferation and increase the spread of waterborne bacteria, including zoonosis agents. Accordingly, the microbiological surveillance of fish used for cosmetic treatment is extremely important, particularly in association with mortality outbreaks.     

Outbreaks of Streptococcosis associated with Streptococcus iniae in Siberian sturgeon (Acipenser baerii) in China

Streptococcus iniae has emerged as an important fish pathogen over the past few decades causing high losses in aquaculture farms all over the world. At least 27 species of fish have been documented to be infected by S. iniae, including cultured and wild populations. In August and October 2013, a serious infectious disease characterized by body ulcer, internal organs haemorrhages and nodules showing on epicardium occurred on the Acipenser baerii farms in Ya'an country, China. Histological examination revealed a multisystemic, necrotising inflammatory response that was particularly marked in liver, kidney, heart and brain. Mass mortality (>40%) was observed in infected fish and two Gram‐positive cocci (Ab130920 and Ab131025) were obtained from kidneys and livers of diseased fish. Experimental infections with these two isolates resulted in marked symptoms in the sturgeons similar to those observed in natural outbreaks, and the LD₅₀ values of the two isolates were 5.1 × 10⁵ and 6.4 × 10⁵ cfu per fish respectively. The two microorganisms were identified as S. iniae through physiological and biochemical tests, 16S rRNA and lctO gene sequence analysis. Both two isolates showed a similar antibiotic susceptibility, which were sensitive to ampicillin, amoxicillin, cefazolin, amikacin, deoxycycline, florfenicol, azithromycin, ciprofloxacin, vancomycin and resistant to streptomycin, gentamicin, kanamycin, norfloxacin and sinomin (SMZ/TMP). Multiplex PCR assay for virulence genes showed both isolates possessed six main virulence genes: simA, scpI, pdi, pgm, cpsD and sagA genes. These results indicated that S. iniae could act as a pathogen of farmed A. baerii. This is the first report of S. iniae infection associated with mass mortality in A. baerii.