A bacterium with close genetic identity to Pseudomonas mandelii associated with spring fish kills in wild bluegill Lepomis macrochirus Rafinesque and pumpkinseed sunfish Lepomis gibbosus (Linnaeus)

Pseudomonas fluorescens are known bacterial pathogens in fish. The P. fluorescens group contains at least nine different bacterial species, although species from fish have rarely been differentiated. Two isolated fish kills affecting wild bluegills, Lepomis macrochirus Rafinesque, and pumpkinseed sunfish, Lepomis gibbosus (Linnaeus), occurred in the spring of 2015 during cool water temperatures (12.5°C–15.5°C). Disease signs included severe bacteraemia with rare gross external signs. Pure bacterial cultures isolated from kidneys of all affected fish were identified as P. fluorescens using the API 20NE system, while no bacteria were isolated from asymptomatic fish. To further identify the species of bacterium within the P. fluorescens complex, genetic analysis of the 16S rRNA, rpoD and gyrB genes was conducted. DNA sequences of bacterial isolates from both mortality events were identical and had close identity (≥99.7%) to Pseudomonas mandelii. Although likely widespread in the aquatic environment, this is the first report of a bacterium closely resembling P. mandelii infecting and causing disease in fish. The bacterium grew at temperatures between 5°C and 30°C, but not at 37°C. It is possible that infections in fish were a result of immunosuppression associated with spring conditions combined with the psychrotrophic nature of the bacterium.     

Coccidiosis due to Goussia lomi (Protista: Apicomplexa) in aquarium-reared Murray cod, Maccullochella peeli (Mitchell), (Percichthyidae)

Abstract. Intestinal parasitism by the coccidian Goussia lomi was associated with morbidity and histological lesions in aquarium-reared Murray cod fry, Maccullochella peeli (Mitchell), at a hatchery near Narrandera, New South Wales, Australia. Coccidial infection was not found in pond-reared Murray cod of the same age, nor in trout cod, M. macquariensis (Cuvier & Valenciennes), reared under similar conditions at the same hatchery. Overcrowding may have predisposed Murray cod in aquaria to heavy infection with G. lomi. Clinical coccidiosis was expressed in a small proportion of infected fish. Large numbers of coccidial developmental forms were detected histologically in the intestinal mucosa of affected fish in association with a necrotic enteropathy. Using a method of intestinal trypsin digestion and flotation, 2000 coccidial oocysts were counted in one 0-05g fish.     

Evaluating potential competitive bottlenecks between invasive common carp and native bluegill and yellow perch

Processes influencing fish recruitment are often highly complex and inherently difficult to understand. Invasive species may complicate recruitment through habitat and food web modifications resulting in competitive bottlenecks. Common carp Cyprinus carpio have been distributed worldwide, and their introductions have resulted in destructive effects on aquatic ecosystems and food web dynamics. Common carp are highly fecund, and high densities of age‐0 carp may occur in some years that may reduce invertebrate prey resources and adversely affect native age‐0 fishes. We used enclosures and field observations to examine potential effects of age‐0 common carp on growth and survival of age‐0 yellow perch Perca flavescens and bluegill Lepomis macrochirus. Yellow perch and bluegill were stocked into enclosures with and without common carp (31 fish/m³) using a substitution experimental design, and fish growth and survival and invertebrate prey resources were assessed. Common carp reduced growth of yellow perch but not bluegill and did not affect survival of either species in mesocosms. Next, we used patterns of common carp, bluegill, and yellow perch abundance and total length across 38 lake‐years to evaluate potential interspecific interactions in natural systems. Age‐0 common carp abundance was not negatively related to size or abundance of bluegill or yellow perch. However, adult common carp and age‐0 yellow perch abundance were inversely related, suggesting a potential competitive bottleneck. Thus, age‐0 common carp may suppress growth of yellow perch when prey is limited, but adult common carp may have larger effects than early life stages on native juvenile fishes.     

Aeromonas salmonicida infection levels in pre‐ and post‐stocked cleaner fish assessed by culture and an amended qPCR assay

Due to increasing resistance to chemical therapeutants, the use of ‘cleaner fish’ (primarily wrasse, Labridae, species) has become popular in European salmon farming for biocontrol of the salmon louse, Lepeophtheirus salmonis (Krøyer). While being efficient de‐licers, cleaner fish mortality levels in salmon cages are commonly high, and systemic bacterial infections constitute a major problem. Atypical furunculosis, caused by Aeromonas salmonicida A‐layer types V and VI, is among the most common diagnoses reached in clinical investigations. A previously described real‐time PCR (qPCR), targeting the A. salmonicida A‐layer gene (vapA), was modified and validated for specific and sensitive detection of all presently recognized A‐layer types of this bacterium. Before stocking and during episodes of increased mortality in salmon cages, cleaner fish (primarily wild‐caught wrasse) were sampled and screened for A. salmonicida by qPCR and culture. Culture indicated that systemic bacterial infections are mainly contracted after salmon farm stocking, and qPCR revealed A. salmonicida prevalences of approximately 4% and 68% in pre‐ and post‐stocked cleaner fish, respectively. This underpins A. salmonicida's relevance as a contributing factor to cleaner fish mortality and emphasizes the need for implementation of preventive measures (e.g. vaccination) if current levels of cleaner fish use are to be continued or expanded.     

Energy allocation patterns in a multiple spawning sunfish: evidence for an income‐based reproductive strategy

Timing of reproduction is an important life‐history trait that varies among species as a function of energy allocation strategy, particularly as individuals are influenced by seasonal variations prior to and during the reproductive season. This study investigated size‐ and sex‐based seasonal patterns of energy allocation in bluegill, Lepomis macrochirus (Rafinesque), a species that spawns repeatedly throughout the summer, by quantifying gonad, liver and mesenteric fat masses. Results indicate that bluegill delay production of gonad tissue until the onset of the spawning season and use an ‘income spawning’ approach. Comparisons of energetic trends with crappies, Pomoxis annularis (Rafinesque) and P. nigromaculatus (Lesueur), a confamilial species of relatively similar size and morphology that spawns in the spring, highlight the flexibility of energy allocation patterns even among relatively similar fishes and shed light on bluegill life‐history strategies.     

Systemic infection in European perch with thermoadapted virulent Aeromonas salmonicida (Perca fluviatilis)

In non‐salmonid fish, Aeromonas salmonicidacan cause local infections with severe skin ulcerations, known as atypical furunculosis. In this study, we present a systemic infection by a virulent A. salmonicidain European perch (Perca fluviatilis).This infection was diagnosed in a Swiss warm water recirculation aquaculture system. The isolate of A.  salmonicida encodes a type three secretion system (TTSS) most likely located on a plasmid similar to pAsa5/pASvirA, which is known to specify one of the main virulence attributes of the species A. salmonicida. However, the genes specifying the TTSS of the perch isolate show a higher temperature tolerance than strains isolated from cold‐water fish. The function of the TTSS in virulence was verified in a cytotoxicity test using bluegill fry and epithelioma papulosum cyprinid cells.     

Co‐infection of rainbow trout (Oncorhynchus mykiss) with infectious hematopoietic necrosis virus and Flavobacterium psychrophilum

Co‐infection of rainbow trout with infections haematopoietic necrosis virus (IHNV) and Flavobacterium psychrophilum is known to occur, and it has been speculated that a combined infection can result in dramatic losses. Both pathogens can persist in fish in an asymptomatic carrier state, but the impact of co‐infection has not been well characterized or documented. In this study, it was hypothesized that fish co‐infected with F. psychrophilum and IHNV would exhibit greater mortality than fish infected with either pathogen alone. To test this, juvenile rainbow trout were co‐infected with low doses of either IHNV or F. psychrophilum, and at 2 days post‐initial challenge, they were given a low dose of the reciprocal pathogen. This combined infection caused high mortality (76.2%–100%), while mortality from a single pathogen infection with the same respective dose was low (5%–20%). The onset of mortality was earlier in the co‐infected group (3–4 days) when compared with fish infected with F. psychrophilum alone (6 days) or IHNV (5 days), confirming the synergistic interaction between both pathogens. Co‐infection led to a significant increase in the number of F. psychrophilum colony‐forming units and IHNV plaque‐forming units within tissues. This finding confirms that when present together in co‐infected fish, both pathogens are more efficiently recovered from tissues. Furthermore, pathogen genes were significantly increased in co‐infected groups, which parallel the findings of increased systemic pathogen load. Extensive tissue necrosis and abundant pathogen present intracellularly and extracellularly in haematopoietic tissue. This was pronounced in co‐infected fish and likely contributed to the exacerbated clinical signs and higher mortality. This study provides novel insight into host–pathogen interactions related to co‐infection by aquatic bacterial and viral pathogens and supports our hypothesis. Such findings confirm that mortality in fish exposed to both pathogens is greatly elevated compared to a single pathogen infection.     

Bluegill Picornavirus isolated from a mortality event involving Bluegill (Lepomis macrochirus) in the upper Mississippi River

A mortality event involving an estimated 1,000 adult bluegills (Lepomis macrochirus) was observed in an ice‐covered backwater lake of the upper Mississippi River near Alma, Wisconsin, in December of 2017. Macroscopic signs of disease included abdominal distension due to fluid accumulation within the internal organs as well as external and internal haemorrhaging. Histological evaluation revealed chronic peritonitis with peritoneal adhesions in all fish examined. Kidney, spleen and ascites fluid samples were collected from diseased bluegills and examined for the presence of pathogens. Bluegill picornavirus (BGPV) was isolated using tissue cell culture methods utilizing a recently developed, uncharacterized bluegill fry cell line (BF‐4), and the presence of this virus was confirmed through molecular identification. The current geographic range, known susceptible hosts as well as historical epizootics associated with BPGV is discussed. The ability of BGPV to cause significant mortality in wild fish further emphasizes the importance of monitoring both wild and hatchery populations for this pathogen.     

Comparative study of the intestinal microbial community of wild and cultured kaluga sturgeon,Huso dauricus

The kaluga, Huso dauricus (Georgi 1775), is a large‐sized semi‐anadromous sturgeon native to the Amur River basin. Gut microbes play an important role in the growth and development of fish. Because wild samples of this species are difficult to obtain, very few studies have analysed the microbial community of the kaluga gut. Here, we used high‐throughput sequencing to determine differences in the intestinal microbial communities of wild and cultured kaluga, to lay the foundation for development of micro‐ecological preparations that might ultimately assist with conserving wild kaluga by benefitting sturgeon aquaculture. Two 2‐year‐old (wild) fish collected from the Heilongjiang River basin were size‐ and age‐matched with two 2‐year‐old (cultured) fish obtained from a breeding base in Yunnan Province, for molecular analysis of gut samples. In total, 483,008 16S rRNA sequences were obtained. The intestinal microbial diversity was greater in the wild kaluga than in the cultured fish. The dominant phyla in the gut community of wild kaluga were Proteobacteria (47.9%), Fusobacteria (30.5%) and Firmicutes (8.85%), whereas samples from the cultured kaluga were dominated by Fusobacteria (78%). This result may be attributed to differences in environmental conditions, particularly water quality and temperature, as well as diet between the wild and cultured animals. The findings provide basic data to assist further research and the development of feed as a part of artificial breeding technology.     

Swimming performance of brown trout and grayling show species‐specific responses to changes in temperature

Fishways have historically been constructed to restore and preserve the ecological connectivity for fish in fragmented rivers. However, the fishways are often selective on species due to different size and swimming capacity. As the proportion of dammed rivers is still increasing, there is a growing need for more information on wild fish and their migration potential. In this study, we compared the swimming capacity of wild caught brown trout (Salmo trutta) and European grayling (Thymallus thymallus) until the fish were exhausted in a critical swimming speed (U_crit) test, under three different naturally occurring stream temperatures in Norway: 1.7, 5.5 and 10°C. The results indicated that trout swim better at the warmer temperatures than at colder temperatures. The grayling showed consistent swimming patterns with little variation across all tested temperatures. The results therefore signify the need to have operational fishways already early in the spring when the grayling migration starts and highlight the need for more studies on fish migration abilities across a wider range of species and seasons.     

Effects of fish species composition on Diphyllobothrium spp. infections in brown trout – is three‐spined stickleback a key species?

Subarctic populations of brown trout (Salmo trutta) are often heavily infected with cestodes of the genus Diphyllobothrium, assumedly because of their piscivorous behaviour. This study explores possible associations between availability of fish prey and Diphyllobothrium spp. infections in lacustrine trout populations. Trout in (i) allopatry (group T); (ii) sympatry with Arctic charr (Salvelinus alpinus) (group TC); and (iii) sympatry with charr and three‐spined stickleback (Gasterosteus aculeatus) (group TCS) were contrasted. Mean abundance and intensity of Diphyllobothrium spp. were higher in group TCS compared to groups TC and T. Prevalence, however, was similarly higher in groups TCS and TC compared to group T. Zero‐altered negative binomial modelling identified the lowest probability of infection in group T and similar probabilities of infection in groups TC and TCS, whereas the highest intensity was predicted in group TCS. The most infected trout were from the group co‐occurring with stickleback (TCS), possibly due to a higher availability of fish prey. In conclusion, our study demonstrates elevated Diphyllobothrium spp. infections in lacustrine trout populations where fish prey are available and suggests that highly available and easily caught stickleback prey may play a key role in the transmission of Diphyllobothrium spp. parasite larvae.     

The Three‐spined Stickleback,Gasterosteus aculeatus Linnaeus 1758, plays a minor role as a host of Lepeophtheirus salmonis (Krøyer 1837) in the Gulf of Maine

The sea louse, Lepeophtheirus salmonis (Krøyer 1837), is a significant parasite of farmed salmon throughout the Northern Hemisphere. Management of on‐farm louse populations can be improved by understanding the role that wild fish play in sustaining and providing refuge for the local population of sea lice. In this study, 1,064 sticklebacks were captured. Of these animals, 176 individuals were carrying a total of 238 sea lice, yielding a prevalence and intensity of 16.5% and 1.4 lice per fish, respectively. Detailed examination of the sea lice on the three‐spined sticklebacks captured in Cobscook Bay found two L. salmonis individuals using three‐spined sticklebacks as hosts. A 2012 survey of wild fish in Cobscook Bay, Maine, found multiple wild hosts for Caligus elongatus (von Nordmann 1832), including three‐spined sticklebacks (Gasterosteus aculeatus L.), but no L. salmonis were found in this earlier study.     

In vitro cultivation model for Heterosporis saurida (Microsporidia) isolated from lizardfish,Saurida undosquamis (Richardson)

Heterosporis saurida is a microsporidian that infects lizardfish, Saurida undosquamis (Richardson, 1848), in the Arabian Sea. Spores were isolated from infected lizardfish and used to infect derived fish cell lines: common carp brain (CCB), epithelioma papulosum cyprinid (EPC), fathead minnow epithelial (FHM), rainbow trout gonad (RTG), bluegill fry (BF‐2) and chinook salmon embryo (CHSE). Non‐fish cell lines were also tested that include: insect (SF‐9), rabbit (RK‐13) and African green monkey (Vero E6). No growth of H. saurida was observed in any fish cell line, SF‐9 or Vero E6 cell lines. H. saurida spores grew only in RK‐13 cell line and were detected by immunofluorescence. Developmental stages of H. saurida were seen in RK‐13 cells by light and transmission electron microscopy, and species identification was confirmed by sequencing. This study demonstrated that H. saurida was able to proliferate in the mammalian RK‐13 cell line, which thus represents an in vitro model for conducting molecular genetics and cell–pathogen interaction studies of Heterosporis.     

Seasonal patterns in the community of gill monogeneans on wild versus cultured orange‐spotted grouper,Epinephelus coioides Hamilton, 1822 in Daya Bay,South China Sea

Epinephelus is among the most important fish genera in the Southeast Asian Pacific coastal systems. In Chinese coast, the orange‐spotted grouper Epinephelus coioides is one of the most significant species based on culture volume, and it is consumed all over the world. Very little information exists on this species’ parasitofauna. Composition and temporal variation in the community structure of the gill monogeneans on wild versus cultured E. coioides from Daya Bay (23°25′N, 117°2′E), South China Sea were determined using seasonal samples taken between April 2008 and January 2009. Eight species of monogeneans of three families and four genera on the gills of E. coioides were found: Neobenedenia melleni, Haliotrema cromileptis, Diplectanum grouperi, Pseudorhabdosynochus justinei, P. lantauensis, P. coioidesis, P. serrani and P. shenzhenensis. Eight of the monogenean species exhibited seasonal variation in their infection dynamics associated with environmental changes during seasons. The variations in the infection dynamics generated changes in the community structure during the sampling periods. Moreover, in the three different host living environments (wild, monocultured and polycultured), the monogenean communities exhibited the different seasonal patterns.     

Ca. Branchiomonas cysticola,Ca. Piscichlamydia salmonis and Salmon Gill Pox Virus transmit horizontally in Atlantic salmon held in fresh water

Elucidation of the role of infectious agents putatively involved in gill disease is commonly hampered by the lack of culture systems for these organisms. In this study, a farmed population of Atlantic salmon pre‐smolts, displaying proliferative gill disease with associated Candidatus Branchiomonas cysticola, Ca. Piscichlamydia salmonis and Atlantic salmon gill pox virus (SGPV) infections, was identified. A subpopulation of the diseased fish was used as a source of waterborne infection towards a population of naïve Atlantic salmon pre‐smolts. Ca. B. cysticola infection became established in exposed naïve fish at high prevalence within the first month of exposure and the bacterial load increased over the study period. Ca. P. salmonis and SGPV infections were identified only at low prevalence in exposed fish during the trial. Although clinically healthy, at termination of the trial the exposed, naïve fish displayed histologically visible pathological changes typified by epithelial hyperplasia and subepithelial inflammation with associated bacterial inclusions, confirmed by fluorescent in situ hybridization to contain Ca. B. cysticola. The results strongly suggest that Ca. B. cysticola infections transmit directly from fish to fish and that the bacterium is directly associated with the pathological changes observed in the exposed, previously naïve fish.     

Genomic comparison of virulent and non‐virulent Streptococcus agalactiae in fish

Streptococcus agalactiae infections in fish are predominantly caused by beta‐haemolytic strains of clonal complex (CC) 7, notably its namesake sequence type (ST) 7, or by non‐haemolytic strains of CC552, including the globally distributed ST260. In contrast, CC23, including its namesake ST23, has been associated with a wide homeothermic and poikilothermic host range, but never with fish. The aim of this study was to determine whether ST23 is virulent in fish and to identify genomic markers of fish adaptation of S. agalactiae. Intraperitoneal challenge of Nile tilapia, Oreochromis niloticus (Linnaeus), showed that ST260 is lethal at doses down to 10² cfu per fish, whereas ST23 does not cause disease at 10⁷ cfu per fish. Comparison of the genome sequence of ST260 and ST23 with those of strains derived from fish, cattle and humans revealed the presence of genomic elements that are unique to subpopulations of S. agalactiae that have the ability to infect fish (CC7 and CC552). These loci occurred in clusters exhibiting typical signatures of mobile genetic elements. PCR‐based screening of a collection of isolates from multiple host species confirmed the association of selected genes with fish‐derived strains. Several fish‐associated genes encode proteins that potentially provide fitness in the aquatic environment.     

Infection and pathology in Queensland grouper,Epinephelus lanceolatus, (Bloch), caused by exposure to Streptococcus agalactiae via different routes

Since 2007, 96 wild Queensland groupers, Epinephelus lanceolatus, (Bloch), have been found dead in NE Australia. In some cases, Streptococcus agalactiae (Group B Streptococcus, GBS) was isolated. At present, a GBS isolate from a wild grouper case was employed in experimental challenge trials in hatchery‐reared Queensland grouper by different routes of exposure. Injection resulted in rapid development of clinical signs including bilateral exophthalmia, hyperaemic skin or fins and abnormal swimming. Death occurred in, and GBS was re‐isolated from, 98% fish injected and was detected by PCR in brain, head kidney and spleen from all fish, regardless of challenge dose. Challenge by immersion resulted in lower morbidity with a clear dose response. Whilst infection was established via oral challenge by admixture with feed, no mortality occurred. Histology showed pathology consistent with GBS infection in organs examined from all injected fish, from fish challenged with medium and high doses by immersion, and from high‐dose oral challenge. These experimental challenges demonstrated that GBS isolated from wild Queensland grouper reproduced disease in experimentally challenged fish and resulted in pathology that was consistent with that seen in wild Queensland grouper infected with S. agalactiae.     

Weaning methods using formulated feeds for snakehead (Channa striata and Channa micropeltes) larvae

The culture of snakehead fish (Channa striata and Channa micropeltes) in Vietnam is limited, and snakehead culture has been banned in Cambodia, because traditional practices include capture of fingerlings from the wild as seed, as well as capture of small‐size (also known as trash fish or low‐value) fish. As hatchery breeding technology has improved, we investigated the optimal weaning practices for these two species. Both laboratory experiments and farm trials were conducted. For C. striata, the optimal weaning procedure is to begin at 17 days after hatch (dah) and wean the fish at 10% replacement of live feed with formulated feed per day. However, for C. micropeltes, the optimal procedure is to wait until 40 dah to begin weaning and then to wean the fish with a 10% replacement of live feed with formulated feed every 3 days. These results should enable farmers to domesticate snakehead culture in Vietnam and Cambodia and eliminate reliance on fish captured from the wild as both seed and feed.     

Effects of release timing on migration behaviour and survival of hatchery‐reared Atlantic salmon smolts in a regulated river

In this study, 221 two‐year‐old hatchery‐reared salmon, Salmo salar, smolts were tagged with radio transmitters over a period of three consecutive years and released in the river in groups of 20–21 fish in various dates between late April and early June. Tagged smolts were tracked during their downstream migration in the lower 36‐km stretch of the regulated River Oulujoki, with the focus on the effects of release date, water temperature and river flow on migration behaviour and survival. The results indicate that release timing and river temperature have profound effects on the initiation of migration, swimming speed and survival of released S. salar smolts. Smolts released early in the spring in cold waters ceased migration after brief downstream movement and were vulnerable to predation, whereas the migration speed and survival rates increased markedly for smolts released later in the spring.