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.     

Prey selection and diets of bluegill Lepomis macrochirus with differing population characteristics in two Nebraska natural lakes

Abstract  Environmental prey samples and stomach contents of bluegill Lepomis macrochirus (Rafinesque) were collected in spring and summer 2000 from two Nebraska Sandhill lakes. Watts Lake contained a low-density bluegill population, whereas Cozad Lake contained a high-density bluegill population. Bluegill diets from both lakes were compared to determine if bluegill prey preference differed between the two populations. The highest median per cent (by calories) of zooplankton in the diet was 1.3%; the remainder was macroinvertebrates. Watts Lake bluegills preferred (based on Manly's alpha) amphipods in spring and chironomids in summer. Cozad Lake bluegills did not show a strong preference (compared with Watts Lake) for any macroinvertebrates, but still utilized amphipods and chironomids during both seasons. Larger bluegills in Watts Lake preferred chironomids in summer, but Cozad Lake bluegills did not exhibit this relationship. The higher density Cozad Lake bluegill population appeared to be more opportunistic than the lower density Watts Lake population, but both preferred macroinvertebrates.     

Replacing fish meal with increasing levels of meat and bone meal,soybean meal and corn gluten meal,in diets of juvenile bluegill,Lepomis macrochirus

A 60‐day study was conducted to determine the response of juvenile bluegill Lepomis macrochirus to seven experimental diets, formulated using a blend of alternative protein sources as a replacement for fish meal. Adequate levels (digestible basis) of energy, protein and amino acids were maintained in diets 1–6, whereas slightly lower protein and energy levels were provided in diet 7. Feed cost per tonne ranged from $ 798.9 (diet 1, 550 g Kg^?1 fish meal) to $ 515.8 (diet 6, 0 g kg^?1 fish meal), or to $ 507.2 (diet 7, 0 g Kg^?1 fish meal). Three commercial diets were included in the study as reference diets: a high‐energy and a low‐energy trout diet, as well as a catfish diet. Quintuplicate bluegill groups (~22 g, n = 10 fish group^?1) were fed the experimental diets twice daily to apparent satiation. No major differences in feed consumption, feed efficiency and growth rates were detected among the bluegill groups fed the experimental diets. Trout diets generally produced higher fish fat deposition, whereas the catfish diet produced a poorer fish growth rate relative to the experimental diets. Under the reported conditions, results indicate diet 6, comprising predominantly soybean meal and porcine meat and bone meal, to be the most economical diet for juvenile bluegill.     

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.     

Analysis of bacterial community and bacterial nutritional enzyme activity associated with the digestive tract of wild Chilean octopus (Octopus mimus Gould, 1852)

Information on the bacterial community associated with octopus is very scarce, unlike fish and other molluscs. This study revealed the bacterial community associated with digestive tract of wild Chilean octopus Octopus mimus using a culture‐dependent method and 16S rDNA clone library. Moreover, we analysed the bacterial nutritional enzyme activity of culturable bacteria. A culture‐dependent method showed that the composition of the culturable bacterial community was substantially different between female and male octopus. The predominant species in female octopus were Vibrionaceae and Streptococcaceae, whereas only Vibrionaceae was dominated in male octopus. Bacterial nutritional enzyme activities of culturable bacteria from male octopus were much higher than female octopus. The 16S rDNA clone library analysis showed that the bacterial community of male octopus exhibited a higher diversity than that of female octopus. The genus Mycoplasma was the predominant bacteria in the digestive tract of all octopus samples. The results obtained in this study raise the possibility that each octopus has different food consumption due to different bacterial community and nutritional enzyme activity, although Mycoplasma sp. is one of the predominant bacteria in the digestive tract. Moreover, our results are useful for the future of microbiological investigation associated with the octopus and for probiotics in the octopus aquaculture.     

Melanotic visceral fibrosis associated with larval infections of Posthodiplostomum minimum and Proteocephalus sp. in bluegill, Lepomis macrochirus Rafinesque, in central Iowa, U.S.A.

Abstract. Severe melanotic liver and visceral fibroses in bluegill associated with infections of plerocercoids of Proteocephalus sp. and metacercariae of the digenean Posthodiplostomum minimum , were studied by light and transmission electron microscopy. Fibrosis was most evident in bluegill carrying heavy worm burdens. Tubular and multilocular melanotic and non-melanotic cysts were formed of concentric layers of non-cellular eosinophilic connective tissue surrounding central spaces containing basophilic material. Moribund and deteriorating plerocercoids and metacercariae were surrounded by, and sometimes invested with, fibrous tissue. Living plerocercoids in the liver were surrounded by necrotic and compressed hepatocyte zones. Fibro-blasts, epithelioid cells and eosinophilic granulocytes were common in compressed cell zones. Intrahepatic fibrotic cysts were surrounded by granulomatous zones. Parenchyma in fibrotic livers showed increased numbers of macrophages, melano-macrophage centres and engorged blood vessels with thickened walls. Histochemical tests for lipofuscin and haemosiderin were strongly positive in melanotic liver tissue. Ultrastructural changes in hepatocytes adjacent to fibrotic cysts and epithelioid zones included intranuclear lipid and glycogen inclusions, chromatin alterations and mitochrondrial degeneration. Bile canaliculi, sinusoids and perisinusoidal spaces showed reduced micro villarsurfacesand a decrease in luminar diameter. Degenerating hepatocytes contained phagolysosomes and myelin bodies.     

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.     

Beef tallow is suitable as a primary lipid source in juvenile Florida pompano feeds

It is assumed that Florida pompano have dietary EPA (20:5n‐3) and DHA (22:6n‐3) requirements. However, it is unclear whether both are equally important in meeting demand for n‐3 long‐chain polyunsaturated fatty acids (LC‐PUFAs) or whether the requirement(s) can be influenced by other fatty acids. Accordingly, we assessed production performance and tissue composition of juvenile Florida pompano (41.0 ± 0.5 g) fed diets containing fish oil; beef tallow; or beef tallow partially or fully supplemented with EPA, DHA or both. After 8 weeks, no signs of fatty acid deficiency were observed. Although fish performance did not vary significantly among the dietary treatments, fish fed the DHA‐supplemented feeds exhibited numerically superior growth than those fed the other diets. Fillets of fish fed the beef tallow‐based diets contained reduced levels of n‐3 fatty acids and LC‐PUFAs and elevated levels of MUFAs and n‐6 fatty acids, although dietary supplementation with EPA and/or DHA attenuated these effects somewhat. Our results suggest that beef tallow is suitable as a primary lipid source in Florida pompano feeds and n‐3 LC‐PUFA requirements may be met by as little as 4 g/kg EPA and 4 g/kg DHA. However, there may be value in supplementing tallow‐based diets with DHA to enhance tissue levels and possibly growth.     

A study on bacteria associated with the intestinal tract of farmed yellow seahorse,Hippocampus kuda (Bleeker, 1852): characterization and extracellular enzymes

The microenvironment of bacteria associated with the gastrointestinal tract of an animal influences the host in many ways, including the metabolism of several nutrients. Isolation, molecular characterization and enzymatic activities of culturable bacteria associated with the gastrointestinal tract of hatchery‐reared oceanic yellow seahorse, Hippocampus kuda, were investigated in the present study. 16S rRNA sequencing of bacterial isolates yielded 10 different bacterial genera belonging to two major groups: proteobacteria (62.9%) and firmicutes (37.03%). The predominant microbial flora belonged to the genera Vibrio spp., Enterovibrio sp. and Bacillus spp. Burkholderia cenocepacia, hitherto unreported from the gastrointestinal tract of marine fish, was retrieved from the seahorse gut, albeit in small numbers. Enzymatic studies indicated that the bacterial isolates possess the ability to degrade lipids, cellulose, xylan, starch and proteins. Lipase activity was exhibited by a majority of the isolates (70.4%), indicating that the bacterial flora associated with the intestinal tract of seahorses is capable of digesting foods rich in lipids. The possible implications for the role of these bacteria in the digestion and physiological processes are discussed. To our knowledge, this is the first time that the characterization and enzymatic activities of bacteria associated with the intestinal tract of farmed seahorses have been reported.     

A spatiotemporal study of bacterial community profiles associated with Atlantic bluefin tuna larvae,Thunnus thynnus L., in three Mediterranean hatcheries

During the first 2 years of larval rearing trials with Atlantic bluefin tuna, survival was a challenging issue. As bacterial colonization of the gut has been shown to play a key role for other species, we studied the profiles of the microbiota associated with individual larvae at different stages in three distant hatcheries. The Bacterial Community Profile (BCP) was quantified based on PCR‐DGGE analyses of partial amplicons from 16S rDNA. Considerable individual variability in BCP was observed before onset of feeding, and the BCP did not show regular fluctuation during ontogenesis. Microalgae were added to the rearing tanks in two of the three hatcheries, but it was not possible to distinguish the effect of location from the effect of algal addition on BCP. In one hatchery, the larvae were reared either with algal addition or in mesocosm, but due to high individual variability, no significant difference in BCP was detected between the two groups. It was hypothesized that this variability was caused by differences in health, physiological status and developmental stage of the larvae. A practical conclusion from the study is the need to analyse a considerable number of individuals to reflect statistically significant differences between the microbial communities associated with rearing groups.     

First isolation of Flavobacterium psychrophilum associated with reports of moribund wild European eel (Anguilla anguilla) in Scotland

In late April 2015, the River Dee Trust informed Marine Scotland Science, Fish Health Inspectorate (FHI), that there had been observations of dead and moribund European eels on the River Dee. Later in May, the Spey Fishery Board also reported a number of moribund European eels in a rotary screw smolt trap on the River Spey. In total, 10 cases involving moribund eels were investigated in 2015 and one case in 2016. In addition, a health screen was conducted to investigate the potential presence of Flavobacterium psychrophilum in healthy eels and Atlantic salmon from the River Dee in 2015. Externally, the diseased eels demonstrated white patches in different locations of the body. In all cases, F. psychrophilum was detected by bacterial isolation and/or molecular methods. Three isolates were further characterized by whole‐genome sequencing (WGS) as belonging to sequence type 15 (ST15). Histological examination of diseased European eels revealed lesions at the level of the integument. The pathogen screen for F. psychrophilum in wild healthy fish tested negative by PCR. Further investigation is required to understand the pathogenicity of this bacterium on the health of eels and the potential impact on the wild salmonid population.     

Potential virulence factors of bacteria associated with tail fan necrosis in the spiny lobster,Jasus edwardsii

Tail fan necrosis (TFN) is a common condition found in commercially exploited spiny lobsters that greatly diminishes their commercial value. Bacteria possessing proteolytic, chitinolytic and lipolytic capabilities were associated with TFN in spiny lobsters, Jasus edwardsii. In this study, 69 bacterial isolates exhibiting all the three enzymatic capabilities from the haemolymph and tail fans of J. edwardsii with and without TFN were further characterized and compared, including morphology, biofilm formation, antimicrobial activity, antimicrobial resistance, and production of siderophores, melanin and ammonia. The genomic patterns of the most common Vibrio crassostreae isolates were also compared between TFN‐affected and unaffected lobsters. Biofilm formation was stronger in bacterial isolates from both haemolymph and tail fans of TFN‐affected lobsters compared to those from the unaffected lobsters, while melanin production and siderophore production were stronger in the isolates from tail fans of lobsters with TFN. By contrast, the other characteristics of isolates were similar in lobsters with and without TFN. The Vib. crassostreae isolates from the affected lobsters had similar genomic patterns. Overall, the results indicate that in addition to proteolytic, chitinolytic and lipolytic activities, the bacteria associated with TFN commonly have enhanced activity of important virulence factors, including biofilm formation, melanin production and siderophore production.     

Water quality and nitrogen budget in turbot Scophthalmus maximus culture system supplemented with lactic acid bacteria

To understand the nutrient utilization efficiency and distribution in indoor fish culture system under treatment of probiotics, this study examined water quality and nitrogen budget in turbot Scophthalmus maximus culture system supplemented with four strains of lactic acid bacteria (LAB) isolated from fishery products. The results showed that the concentration of ammonia nitrogen (NH?‐N) and nitrite nitrogen (NO?‐N) in LAB treatments were all significantly lower (p < 0.05) than in the control group. The nitrate nitrogen (NO?‐N) in LAB treatments showed no significant differences (p > 0.05) compared with control group. The fish feed and water exchange accounted for 79.07% and 17.02% of total N input respectively. For N output, the drainage and residual diets in LAB treatment accounted for 24.50%–25.80% and 1.33%–1.60% respectively, and they were significantly lower than in the control group (27.60% and 2.20% respectively). Fish growth and lost N in LAB treatments accounted for 27.10%–30.50% and 11.00%–18.50% respectively, and they were both significantly higher (p < 0.05) than in the control group (22.30% and 5.30% respectively). The results indicated that the indigenous LAB strains were capable of improving fish growth, and reducing NH?‐N and NO?‐N level (at concentration of 10⁵ cfu/ml) by directly adding in S. maximus culture water. Moreover, specific strains of LAB may increase nitrogen loss by promoting denitrification process in culture system.