Expanded geographical distribution of Myxobolus cerebralis: first detections from Alaska

The parasite responsible for salmonid whirling disease, Myxobolus cerebralis, was introduced to the USA in 1958. It has since spread across the country causing severe declines in wild trout populations, but has never been documented from Alaska. However, while assessing the risk of introduction of M. cerebralis into the state, we detected the parasite using a species-specific polymerase chain reaction (PCR) assay. Testing of 180 hatchery rainbow trout, Oncorhynchus mykiss (Walbaum), by pepsin trypsin digest (PTD) and quantitative PCR (QPCR) revealed 14 positive samples. Infection was confirmed by sequencing the parasite 18S rRNA gene and by a nested PCR assay based on the same gene. Sequence comparison of M. cerebralis from several locations demonstrated the Alaska isolates were genetically distinct and therefore not false-positives arising from contamination during processing. We were unable to visually identify myxospores, indicating that either infection was light or mature spores had not formed. A reference set of fish samples spiked with known numbers of myxospores verified the QPCR and PTD results. This paper presents DNA sequence data from the Alaska M. cerebralis isolates, provides a brief history of the fish and facility of origin, and discusses implications of different testing methods on asymptomatic fish populations.     

Characterization of glycans in the developmental stages of Myxobolus cerebralis (Myxozoa), the causative agent of whirling disease

Glycans and sugar-binding molecules (lectins) form an interactive recognition system, which may enable parasitic organisms to adhere to host cells and migrate into target tissues. The aim of the present study was to analyse surface-associated glycans in the developmental stages of Myxobolus cerebralis (Hofer), the causative agent of whirling disease. A panel of biotin-labelled plant lectins was used to detect a broad spectrum of glycan motifs with high specificity. Binding sites were detected histochemically in the tissue sections of infected rainbow trout, Oncorhynchus mykiss (Walbaum), and infected Tubifex tubifex (Müller), and were characterized by light, fluorescence and transmission electron microscopy. With mannose-specific lectins [Lens culinaris agglutinin, Pisum sativum agglutinin, Canavalia ensiformis agglutinin (LCA, PSA, CanA)] mannose-containing glycans were detected in all the developmental stages and host tissues. No binding sites for galactose-specific lectins were present in M. cerebralis spores but reactivity with host tissues occurred. Diversity in glycans was detected by N-acetyl-D-galactosamine-specific lectins in sporoplasm cells of M. cerebralis and triactinomyxon spores. In the group of lectins with monosaccharide-specificity for N-acetyl-D-glucosamine (GlcNAc), the reactivity of Datura stramonium agglutinin (DSA), Lycopersicon esculentum agglutinin (LEA) and Solanum tuberosum agglutinin (STA) was restricted to polar capsules whereas Griffonia simplicifolia agglutinin II (GSA II) also bound to sporoplasm cells of stages in the fish host but not in those present in infected T. tubifex. Moreover, Triticum vulgaris (wheat germ) agglutinin (WGA) and succinylated WGA indicated the presence of N-acetyl-D-glucosamine polymers in polar capsules. No specificity for spores was observed concerning 'bisected'N-glycans and no reactivity in parasitic stages was observed with the fucose-binding lectin Ulex europaeus agglutinin (UEA) I, Sambucus nigra agglutinin (SNA) (specific for alpha2,6-sialylated glycans) and Maackia amurensis agglutinin (MAAI) (specific for alpha2,3-sialylated glycans). Arachis hypogaea (peanut) agglutinin (PNA), Erythrina cristagalli agglutinin (ECA), GSA I, Sophora japonica agglutinin (SJA), Dolichos biflorus agglutinin (DBA) and GSA II detected reactive sites solely confined to the developmental stages of M. cerebralis and were not reactive in the fish host. These parasite-specific glycans may play a role in the adhesion process of the parasite to fish epidermis prior to infection, but may provide protection to the host by activating the complement system, or stimulating an adaptive immune response as putative antigens.     

Dietary taurine supplementation in plant protein based diets do not affect growth and reproductive performance of zebrafish

Taurine (Tau) has been regarded as a conditional essential nutrient for some fish species. Although its role has been extensively studied in higher vertebrates, limited results are reported with fish especially its role on reproductive performance and the ontogenic changes on Tau levels throughout the life cycle. Therefore, we designed a feeding trial using zebrafish as a model species to test whether Tau supplementation to plant protein diets would have a positive effect on growth and reproductive performance. Zebrafish were fed plant protein diets containing graded levels of Tau (0.2, 4.6, 5.9 and 13.7 g/kg diet) from 10 days post fertilization (dpf) to sexual maturity. An additional commercial diet was used as a positive control for performance. The trial followed a completely randomized design with five treatments (diets) and three replications. After 60 days of feeding, growth, Tau concentration in the body, redox status, lipid body composition, reproductive and offspring performances were analysed. Tau supplementation did not affect growth and/or reproductive performance; however, zebrafish seems to differently modulate Tau concentration according to the growth stage. Tau seemed to induce a hypolipidemic effect in zebrafish by reducing lipid accumulation in their bodies (p < .05). A trend to a more pro‐oxidant effect of Tau supplementation was observed by the decreased reduced glutathione levels. In sum, Tau does not affect growth and reproductive performance of zebrafish but it is important for normal lipid utilization and redox status.     

Anaemia and plasma lipid profile in common carp (Cyprinus carpio) exposed to ambient copper sulphate and nano‐scale copper oxide

The objective of this study was to determine the effects of copper sulphate (CS) and copper oxide nanoparticles (NCuO) on haematological characteristics and plasma lipid profile in Cyprinus carpio. Fish were exposed to CS (0.25 mg L^?1 Cu) and NCuO [0.25 (LNP) and 25 mg L^?1 (HNP) Cu] for 14 days. CS and LNP treatments showed hypochromic anaemia after 3 and 7 days, and macrocytosis and haemolytic anaemia after 14 days. HNP treatment showed hypochromic anaemia and polycythemia after 3 days, and macrocytosis and hypochromic anaemia after 7 days and haemolytic hypochromic anaemia after 14 days. All copper‐exposed fish showed leukopenia at all times, except for HNP after 7 days and CS after 3 days. CS treatment at all times, and NCuO treatments after 14 days showed elevated plasma bilirubin. CS (3rd and 7th days) and HNP (3rd day) treatments had significantly higher plasma triglyceride compared to the control. Significant increases in plasma cholesterol and lipoprotein levels were observed after 3 and 14 days in CS, and after 14 days in LNP treatment; whereas HNP treatment had significantly higher high density lipoprotein (HDL) and very low‐density lipoprotein (VLDL) levels after 3 days, and cholesterol, HDL and low‐density lipoprotein (LDL) after 14 days. In conclusion, CS and NCuO induce anaemia probably due to iron metabolism disorder and RBC destruction in carp. CS and NCuO (at high concentration) alter plasma lipid profile, which may be due to stress‐induced lipolysis and hepatobiliary damages. At similar concentrations, copper nanoparticles are less toxic to carp than copper ions.     

Purified deoxynivalenol or feed restriction reduces mortality in rainbow trout,Oncorhynchus mykiss (Walbaum), with experimental bacterial coldwater disease but biologically relevant concentrations of deoxynivalenol do not impair the growth of Flavobacterium psychrophilum

Diets containing deoxynivalenol (DON) were fed to rainbow trout Oncorhynchus mykiss (Walbaum) for 4 weeks followed by experimental infection (intraperitoneal) with Flavobacterium psychrophilum (4.1 × 10⁶ colony‐forming units [CFU] mL⁻¹). Mortality of rainbow trout fed either 6.4 mg kg⁻¹ DON or trout pair‐fed the control diet was significantly reduced (P < 0.05) in comparison with trout fed the control diet to apparent satiation (<0.1 mg kg⁻¹ DON). In a second experiment, trout were fed one of three experimental diets; a control diet, a diet produced with corn naturally contaminated with DON (3.3 mg kg⁻¹ DON) or a diet containing purified DON (3.8 mg kg⁻¹); however, these fish were not experimentally infected. The presence of DON resulted in significant reduction (P < 0.0001) in feed intake as well as weight gain after 4 weeks. Respiratory burst of head‐kidney leucocytes isolated from rainbow trout fed diets containing purified DON (3.8 mg kg⁻¹) was significantly higher (P < 0.05) at 35 day post‐exposure compared with controls. The antimicrobial activity of DON was examined by subjecting F. psychrophilum in vitro to serial dilutions of the chemical. Complete inhibition occurred at a concentration of 75 mg L⁻¹ DON, but no effect was observed below this concentration (0–30 mg L⁻¹).     

Effects of dietary inorganic copper on growth performance and immune responses of juvenile beluga,Huso huso

A 12‐week feeding trial was conducted to evaluate the optimum dietary inorganic copper (copper sulphate) in juvenile beluga, Huso huso. Eight semi‐purified diets containing 1.1 (Cu_1.0), 3.5 (Cu_4.0), 7.1 (Cu_7.0), 9.7 (Cu₁₀), 13.1 (Cu₁₃), 25.1 (Cu₂₅), 49.9 (Cu₅₀) and 195 (Cu₁₉₅) mg Cu kg^?1 diet in the form of CuSO₄.5H₂O were fed to fish of initial body weight 8.49 ± 0.32 g and length 11.85 ± 0.66 cm (mean ± SD) in triplicate groups in a flow‐through system. Weight gain (WG) of fish fed Cu₁₀ and Cu₁₃ diets was significantly higher than that of fish fed Cu_1.0, Cu_4.0, Cu₂₅, Cu₅₀ and Cu₁₉₅ diets (P < 0.05). Whole‐body and muscle crude protein increased with dietary Cu up to the supplementation level of 13.1 mg kg^?1 diet and then decreased. Whole‐body lipid content was negatively correlated, while whole‐body ash was positively correlated with dietary copper concentration. Hepatic copper–zinc superoxide dismutase activity of fish fed Cu₁₀ and Cu₁₃ diets was significantly higher than that of fish fed Cu_1.0, Cu_4.0 and Cu₁₉₅ diets. Hepatic thiobarbituric acid‐reactive substances of fish fed Cu₁₃ diet was significantly lower than those of fish fed the other diets except for that of fish fed Cu₁₀ diet. Aspartate aminotransferase, alanine aminotransferase and copper accumulation in tissues increased with dietary copper. Broken‐line analysis of WG suggested that the optimum dietary Cu level was 10.3 mg Cu kg^?1 diet. Therefore, these results may indicate that the optimum dietary Cu levels could be greater than 10.3 mg Cu kg^?1 diet but less than 13.1 mg Cu kg^?1 diet in juvenile beluga, when copper sulphate is used as the dietary source of inorganic copper.     

Evaluation of a 4‐h static copper sulphate treatment against experimental infection of Flavobacterium columnare in channel catfish (Ictalurus punctatus)

The efficacy of copper sulphate (CuSO₄) against the early stages of an experimental acute infection of Flavobacterium columnare in channel catfish (Ictalurus punctatus) was evaluated. Fish were challenged, by waterborne exposure to F. columnare in an ultra‐low flow‐through water delivery system, and treated with CuSO₄ at 4.2 and 2.1 mg L^?1 at 5.5 h post challenge for 4 h. Bacterial challenged non‐treated fish acted as a positive control and non‐challenged non‐treated fish acted as a negative control. Fish challenged with F. columnare and treated with CuSO₄ at 4.2 and 2.1 mg L^?1 post challenge had mortalities of 6.4% and 18.3%, respectively, compared with the positive control, which had 90.2% mortality. The mortality of challenged fish treated with CuSO₄ at 4.2 and 2.1 mg L^?1 was significantly different from the positive control. There was no significant difference between the mortalities of the 4.2 and 2.1 mg L^?1 treatments. The results suggest that CuSO₄ has clear therapeutic value against columnaris infections.     

Dose‐dependent protective effects of dietary selenium on abalone Haliotis discus hannai Ino against the toxicity of waterborne copper

This study was designed to assess the protective effects of dietary selenium (Se) on abalone Haliotis discus hannai Ino against the toxicity of waterborne copper (Cu). A 60‐day feeding trial was conducted in a static water system for abalone (initial weight: 3.17 ± 0.01 g) exposed to 0.02 mg L^?1 of waterborne Cu. The animals were fed one of the three experimental diets with 0.10, 1.31 and 4.20 mg kg^?1 of Se from Na₂SeO₃·5H₂O respectively. Results showed that the abalone fed 1.31 mg kg^?1 of dietary Se had the lowest Cu concentration in shell, muscle, mantle, gill, hepatopancreas and serum. Meanwhile, the significant lowest contents of malondiadehyde and protein carbonyl in hepatopancreas were also found in the treatment with 1.31 mg kg^?1 of dietary Se (P < 0.05). In addition, this treatment had significant higher glutathione content and thioredoxin reductase activity in hepatopancreas (P < 0.05). However, the activity of Se‐dependent glutathione peroxidase (Se‐GPx) was significantly decreased in the treatment with 4.20 mg kg^?1 of dietary Se (P < 0.05). In this treatment, the protein carbonyl content in hepatopancreas was significantly higher than that in the group with 1.31 mg kg^?1 of dietary Se (P < 0.05). In conclusion, in terms of anti‐oxidation and Cu accumulation, the protective effects of dietary Se on abalone against waterborne Cu were dose‐dependent. The 1.31 mg kg^?1 of dietary Se had this effect, but not 4.20 mg kg^?1 of dietary Se. Moreover, the latter increased the oxidative stress in abalone exposed to the waterborne Cu.     

Quantitative dietary copper requirement of juvenile Siberian sturgeon,Acipenser baerii,and effects on muscle composition and some enzymatic activities

An 8‐week feeding trial was conducted to quantify dietary copper (Cu) requirement of juvenile Siberian sturgeon, Acipenser baerii. Five isonitrogenous diets were formulated to provide actual dietary copper values of 1.8, 5.7, 10.1, 15.9 and 28.3 mg Cu per kg diet. Experimental diets were fed to the Siberian sturgeon (27.57 ± 0.24 g) in triplicate to apparent satiation for 8 weeks. At the end of experiment, weight gain (WG), specific growth rate (SGR) and protein efficiency ratio (PER) were significantly increased with increasing dietary Cu level up to 10.1 mg/kg and then decreased with further increases in dietary Cu level (p < .05). The Cu concentration in the liver and cartilage was positively correlated with the respective concentrations in the diet (p < .05), while muscle and serum Cu concentrations remained significantly unchanged (p > .05). Superoxide dismutase and glutathione peroxidase had the highest activities in serum of fish fed with 15.9 and 28.3 mg Cu per kg diet, respectively. Analysis by the broken‐line regression of SGR, crude protein content and superoxide activity demonstrated that the optimum dietary Cu requirements in juvenile Siberian sturgeon were 9.51, 9.58 and 16.10 mg/kg diet, respectively.     

Effects of various dietary levels of copper hydroxychloride on growth performance and tissue response in Pacific white shrimp Litopenaeus vannamei fed practical diets

Two trials were conducted to evaluate the performance of Pacific white shrimp Litopenaeus vannamei offered diets containing various copper (Cu) levels from Cu hydroxychloride (Cu₂(OH)₃Cl) containing 58.81% copper in the clear water recirculating system. In both trials, the basal diet (360 g kg^?1 protein, 80 g kg^?1 lipid) containing approximately 10 mg Cu kg^?1 was primarily comprised of fishmeal, soybean meal, corn protein concentrate and whole wheat. In trial 1, test diets were produced supplementing the basal diet with 5, 10, 20, 40 and 60 mg Cu kg^?1 from Cu hydroxychloride. Four replicate groups of 15 shrimp per tank (initial weight 0.28 g) were offered diets in slight excess over 8 weeks. In trial 2, the basal diet was supplemented with 30, 90, 150, 210 and 270 mg Cu kg^?1 from Cu hydroxychloride. Seven replicate groups of 15 shrimp per tank (initial weight 0.22 g) were offered feed in slight excess over 7 weeks. At the end of the two growth trials, no significant differences were observed in final biomass, final mean weight, percentage weight gain, feed conversion ratio (FCR) and survival. In trial 1, the Cu concentrations of the carapace, hepatopancreas and whole shrimp linearly increased with increasing dietary Cu supplements. In trial 2, polynomial regression of Cu concentrations of the carapace, hepatopancreas and whole shrimp against analysed dietary Cu content indicated that a plateau was reached at 215 mg analysed Cu kg^?1. Results of this study indicate that there was no negative effect of high levels of Cu supplement with regard to growth and survival. Tissue levels generally increased up to around 200 mg Cu kg^?1 diet and then decreased, possibly indicating a shift in physiology.     

Dietary copper requirement of fingerling Channa punctatus (Bloch) based on growth,feed conversion,blood parameters and whole body copper concentration

A 12‐week feeding trial was conducted to estimate the dietary copper requirement of fingerling Channa punctatus. Six casein?gelatin‐based test diets (450 g kg^?1 crude protein; 18.81 kJ g^?1 gross energy) with graded levels of copper as copper sulphate (3.7, 4.7, 5.7, 6.7, 7.7 and 8.7 mg copper equivalent kg^?1 diet) were formulated and fed to triplicate groups of fish (7.25 ± 0.81 cm; 5.21 ± 0.27 g) near to satiation. Fish fed diet with 6.7 mg kg^?1 copper had highest absolute weight gain (AWG; 51.63 g fish^?1), protein efficiency ratio (PER; 1.42 g fish^?1), protein gain (PG; 8.34 g fish^?1), haemoglobin (Hb; 9.68 g dL^?1), haematocrit (Hct; 31.18%) and RBCs (3.24 × 10⁶ × mm^?3). Feed conversion ratio (FCR) was found to be best (1.57) at above level of dietary copper. Whole body copper concentration was found to increase with the increasing levels of dietary copper. Hepatic thiobarbituric acid‐reactive substances concentration was found to decrease with increasing dietary concentrations of copper up to 6.7 mg kg^?1 beyond which a reverse trend in this parameter was noted. Broken‐line regression analysis of AWG, FCR and PG concentrations against varying levels of dietary copper yielded the requirement in the range of 6.66–6.78 mg kg^?1. Data generated during this study would be useful in formulating copper‐balanced commercial feeds for the intensive culture of this fish.     

Examination of a practical method for copper enrichment of euryhaline rotifers (Brachionus plicatilis) as diet of Eriocheir sinensis zoea larvae

In this study, an effective method to enrich the rotifer Brachionus plicatilis with copper was developed as a feed for the Chinese mitten crab Eriocheir sinensis zoea larvae. Changes in the concentrations of other minerals in rotifers were also examined when copper was added for rotifer enrichment. The ability of Chlorella to absorb waterborne copper is much higher than that of rotifers, and hence, copper was preaccumulated in Chlorella before its ingestion by rotifers. The copper content in rotifers was comparable to the dietary copper requirement of the crab larvae when the rotifers were enriched with 0.1 mg Cu g⁻¹ Chlorella for 12 h. Further enrichment in rotifers with Cu‐enriched Chlorella and lipid emulsions did not significantly change the profile of major fatty acids and mineral composition in the rotifers. Evidence shows the feasibility of copper enrichment in rotifers using microalgae that can accumulate copper. This study indicates that copper in rotifers can be enriched by feeding copper‐enriched algae at a concentration of 0.1–0.2 mg Cu g⁻¹ Chlorella. The developmental rates of E. sinensis can be improved by feeding zoea larvae with copper‐enriched rotifers, but survival rates were not affected by dietary copper enrichment.     

Dietary ratios of n‐3/n‐6 fatty acids do not affect growth of Nile tilapia at optimal temperatures (28°C) nor at temperatures that simulate the onset of winter (22°C)

Nile tilapia (Oreochromis niloticus) juveniles were fed diets containing 13 g/kg total polyunsaturated fatty acids (PUFAs) at different n‐3/n‐6 dietary ratios (0.2, 0.5, 0.8, 1.3 and 2.9) for 56 days, at 28°C. Subsequently, fish were submitted to a winter‐onset simulation (22°C) for 33 days. PUFA n‐3/n‐6 dietary ratios did not affect fish growth at either temperature. At 28°C, tilapia body fat composition increased with decreasing dietary PUFA n‐3/n‐6. Winter‐onset simulation significantly changed feed intake. The lowest dietary n‐3/n‐6 ratio resulted in the highest feed intake. At both temperatures, body concentrations of α‐linolenic acid, docosahexaenoic acid, eicosatrienoic acid and docosapentaenoic acid decreased as dietary n‐3/n‐6 decreased. Body concentrations of eicosapentaenoic acid (EPA, 20:5 n‐3) increased with decreasing concentrations of dietary EPA. The n‐6 fatty acids with the highest concentrations in tilapia bodies were linoleic acid and arachidonic acid (ARA, 20:4 n‐6). At 28°C, SREBP1 gene expression was upregulated in tilapia fed the lowest n‐3/n‐6 diet compared to tilapia fed the highest n‐3/n‐6 ratio diet. Our results demonstrate that a dietary PUFA of 13 g/kg, regardless of the n‐3/n‐6 ratio, can promote weight gains of 2.65 g/fish per day at 28°C and 2.35 g/fish per day at 22°C.     

Effect of repeated exposure to AQUI‐S® on the viability and growth of Neoparamoeba perurans

There have been recent efforts amongst immunologists to develop approaches for following individual fish during challenges with viral and bacterial pathogens. This study contributes to assessing the feasibility of using such approaches to study amoebic gill disease (AGD). Neoparamoeba perurans, agent of AGD, has been responsible for widespread economic and fish loss in salmonid aquaculture. With the emergence of AGD in Europe, research into infection dynamics and host response has increased. This study investigated the effect of repeat exposure to anaesthesia, a necessary requirement when following disease progression in individual fish, on N. perurans. In vitro cultures of N. perurans were exposed every 4 days over a 28‐day period to AQUI‐S^® (isoeugenol), a popular anaesthetic choice for AGD challenges, at a concentration and duration required to sedate post‐smolt salmonids. Population growth was measured by sequential counts of amoeba over the period, while viability of non‐attached amoeba in the culture was assessed with a vital stain. AQUI‐S^® was found to be a suitable choice for in vivo ectoparasitic challenges with N. perurans during which repetitive anaesthesia is required for analysis of disease progression.     

Effect of different culture conditions on the structural diversity of prokaryote communities in the sediment of earth ponds stocked with gilthead seabream Sparus aurata (Linnaeus, 1758)

In this study, we evaluate the use of polymerase chain reaction‐denaturing gradient gel electrophoresis (PCR‐DGGE) for monitoring the effect of different aquaculture practices on sediment prokaryote (Archaea and Bacteria) communities. The effect of initial fish (gilthead seabream Sparus aurata) stocking density on the structural diversity of prokaryote communities of earth ponds bottom sediments was evaluated using PCR‐DGGE after a 5 month grow‐out period. An identical approach was used to monitor the effect of supplying different fish feeds [commercial feed (CD) versus an ecofeed (ECO)]. One additional variable was the use of copper sulphate (CuSO₄) as an algicide in some of the experimental rearing tanks. The statistical analyses of prokaryote community profiles showed that the presence of fish in earth ponds significantly influenced the structure of sediment prokaryote communities, when compared with earth ponds without fish, independently of the stocking density. Our results also indicated that the structure of the prokaryote communities of earth ponds supplied with the ECO feed shared a strong similarity with that fed CD. Curiously, the use of CuSO₄ in ponds receiving the ECO feed promoted significant differences on the structural composition of the bacterial community, but not on the archaeal community. DGGE molecular fingerprints are suitable for fast evaluation of new management practices in food‐fish production on earth ponds by monitoring shifts on microbial communities in bottom sediments.     

Flavobacteria colonizing the early life stages of hatchery‐incubated Chinook salmon Oncorhynchus tshawytscha (Walbaum 1792) are markedly diverse

Flavobacterial diseases are significant impediments to hatchery‐based fishery conservation and aquaculture productivity worldwide. Recent studies revealed a multitude of novel flavobacteria within the reproductive fluids and unfertilized eggs of feral Chinook salmon Oncorhynchus tshawytscha broodstock, some of which were associated with systemic disease. Herein, embryonated eggs/fry from these broodstock were assayed for flavobacteria while in incubator stacks in three hatcheries over 2 years, as was the water entering hatchery incubators. Overall, >65% of sampled eggs and 38% of fry were colonized by flavobacteria. One hundred and ninety‐one egg and fry‐associated flavobacterial isolates were characterized phenotypically and via 16S rRNA gene sequencing and phylogenetic analyses, revealing that the majority fell into 22 clades (i.e., 15 Flavobacterium spp. groups and seven Chryseobacterium spp. groups) that varied in presence by facility. Although some matched previously described fish‐pathogenic species, the majority were distinct from all described flavobacteria and likely represent novel species. Of concern, iodophor disinfection at the commonly utilized dose/duration for egg‐surface disinfection did not eliminate flavobacteria. Results also implicated maternal routes of infection and source water for some flavobacteria. In total, study findings underscore the complexity of flavobacterial ecology within hatchery environments and highlight the need for improved hatchery biosecurity practices.     

Effects of dietary inclusion of various concentrations of Scutellaria baicalensis Georgi extract on growth,body composition,serum chemistry and challenge test of far eastern catfish (Silurus asotus)

Effects of various concentrations of Scutellaria baicalensis (SB) extract in diets on growth, body composition, serum chemistry and disease challenge test of far eastern catfish (Silurus asotus) were determined and compared with a commercially available immune enhancer. Eight experimental diets were prepared in triplicate: control (Con) diet without supplementation of SB and SB‐0.25, SB‐0.5, SB‐1, SB‐2, SB‐3 and SB‐5 diets containing SB at concentrations of 0.25, 0.5, 1, 2, 3 and 5%, respectively. In addition, 0.1% of a commercial immune enhancer product (CP) was also tested. No significant difference in weight gain of fish was found. Feed consumption, feed efficiency ratio and protein retention of fish were not affected by the experimental diets. At the end of the 8‐week feeding trial, 10 externally normal fish from each tank were infected by Vibrio anguillarum or Strepotococcus iniae. Cumulative mortality of fish fed the Con diet was higher than that of fish fed the all other diets in 10 and 25 days after V. anguillarum or S. iniae infection. Results of this study indicated that dietary inclusion of SB extract was effective in improving survival of eastern catfish after V. anguillarum and S. iniae infection, but the various concentrations of SB did not affect fish performance.     

Recovery of Bacillus mycoides,B. pseudomycoides and Aeromonas hydrophila from common carp (Cyprinus carpio) and rainbow trout (Oncorhynchus mykiss) with gill disease

During a 3‐month period from June to the end of August 2016, ~5% mortalities were observed in a farm with rainbow trout (Oncorhynchus mykiss Walbaum) and one farm of common carp (Cyprinus carpio L.) in Bulgaria. The disease was manifested by gill ulcers/rot, asphyxiation and bloody ascites. Aeromonas hydrophila was isolated from the internal organs of all the diseased fish. Bacillus mycoides or B. pseudomycoides were recovered from the gill lesions on diseased carp and rainbow trout, respectively, with identification achieved by conventional phenotyping and by sequencing of the 16S rRNA gene. In vivo experiments confirmed that all three organisms were pathogenic to rainbow trout.