Vibrio harveyi and other bacterial pathogens in cultured summer flounder, Paralichthys dentatus

A monitoring program for Vibrio harveyi and other potential bacterial pathogens of summer flounder was conducted at two facilities in the Northeast United States. Bacterial samples were collected from larval and juvenile fish and live-feed, and identified using API 20E biochemical profiles and 16S rDNA sequencing. Histopathological examinations were conducted in order to relate histological changes with the presence of potential bacterial pathogens. V. harveyi, Vibrio ichthyoenteri, and Photobacterium damselae subsp. damselae, three known pathogens of flatfish, were isolated from diseased summer flounder. Although a high proportion of juvenile summer flounder showed microscopic signs of disease, the presence of these potential bacterial pathogens in fish was not associated with large-scale mortalities. An outbreak of flounder infectious necrotizing enteritis with 30% cumulative mortality occurred when juveniles were transported to a new facility. Isolates of V. harveyi from the disease outbreak were genetically different from the isolates from the commercial hatchery. V. harveyi isolates from both facilities were pathogenic to summer flounder by intraperitoneal injection. P. damselae subsp. damselae, V. ichthyoenteri, and Vibrio scophthalmi, also found in fish with gross lesions, were not pathogenic to juvenile summer flounder by intraperitoneal injection. Our research shows that several potential bacterial pathogens are associated with morbidity and mortality in summer flounder larvae and juveniles, especially in situations of stress. Increased knowledge about the environmental conditions that lead to disease, as well as the interactions of V. harveyi with other microbial species, could lead to the development of management strategies in summer flounder farms.     

The effects of ozone and probiotics on the survival of black tiger shrimp (Penaeus monodon)

Total ozone production (TOP) from an ozonator, residual ozone concentration (ROC) in water, and the effects of ozone with or without probiotic supplemented feeds on bacterial growth, and shrimp (Penaeus monodon) survival were investigated. Minimal effective ROC to inhibit 3 log units of Vibrio harveyi D331 for 6 h and 2 log units of Bacillus S11 for 9 h was 0.38 g O₃/l of ROC from 5-min ozonation. Shrimp postlarvae exposed to 0.34–0.50 mg O₃/l ROC (8-h ozonation) caused loss of balance, immobility and destruction of gill lamellar epithelium. In vivo treatment of juvenile P. monodon reared on probiotic feed for 1 month revealed that 0.35 mg O₃/l ROC (30-min ozonation) effectively inhibited 3 log units of V. harveyi D331 for 24 h. At this ROC dosage, there was no effect either on shrimp or on intestinal probiotics (Bacillus S11). Shrimp survival from probiotic treatment, coupling with ozonation, increased significantly (P≤0.05) compared with controls.     

Effect of water temperature on the immune response of white shrimp Litopenaeus vannamei to Vibrio alginolyticus

White shrimp Litopenaeus vannamei held in 25‰ seawater at 27 °C or 28 °C were injected with TSB-grown Vibrio alginolyticus at 1 × 10⁴ colony-forming units (cfu) shrimp^− 1 or 1 × 10⁵ cfu shrimp^− 1, and then cultivated onward at water temperatures varying from 20 to 34 °C. Over 24–144 h, mortality of V. alginolyticus-injected shrimp held at 34 °C or 32 °C was significantly higher than that of shrimp held at lower temperatures. In a separate experiment, shrimp held in 25‰ seawater at 28 °C and then cultured onward at 20 to 32 °C were examined for immune parameters at 24–96 h. THC, phenoloxidase activity, respiratory burst, and SOD activity decreased significantly at 24 h after transfer to 32 °C. Shrimp held in 25‰ seawater at 27 °C and then cultured onward at 20 to 34 °C showed a significant reduction in phagocytic activity and clearance efficiency for V. alginolyticus at 24 h after transfer to 34 °C. It was concluded that transfer of shrimp from 27 or 28 °C to higher temperatures (32 and 34 °C) reduced their immune capability and decreased resistance to V. alginolyticus infection.     

Effects of varying Artemia enrichment on growth and survival of juvenile seahorses, Hippocampus abdominalis

Seahorses are the focus of recent aquaculture ventures in a variety of countries. Many of these ventures utilise enriched live Artemia (to varying degrees) to feed their seahorses. Given the range of commercially available enrichment products that can be used to enrich Artemia, it is imperative to determine which of these may promote the best growth and survival in seahorses, as well as being most cost-effective. This investigation tested the effects on juvenile seahorse (Hippocampus abdominalis Leeson, 1827) growth and survival through enriching Artemia with three commercial enrichment products: Super Selco®, DHA Protein Selco® and Algamac-3050. These were tested against a low-cost Artemia on-growing mixture used in this instance as an enrichment (90% Eyre Peninsula Aquaculture brine shrimp food (EPABSF)/10% spirulina).

After 3 months, there was a significant difference in juvenile length between the enrichment treatments, with juveniles in the DHA Protein Selco® and Algamac-3050 treatment longer than juveniles in the Super Selco® treatment, but not longer than juveniles in the EPABSF/spirulina treatment. There was also a significant difference in juvenile wet weights between the treatments, with juveniles in the Super Selco® treatment weighing significantly less than juveniles in the other three treatments.

In terms of condition factor (CF), there was a slight significant difference between the treatments, with juveniles in the DHA Protein Selco® and EPABSF/spirulina treatment having higher CFs than juveniles in the Super Selco® treatment, but not the Algamac-3050 treatment. Mean daily specific growth rate (SGR) for Super Selco® was lower than the other three treatments. There was no effect of enrichment treatment on seahorse survival with 100% survival in all four treatments.

Fatty acid analysis revealed marked differences in the Artemia enriched with the various enrichment products. Super Selco®- and Algamac-3050-enriched Artemia had the highest percentage of docosahexanoic acid (DHA), 22:6(n−3), while Super Selco®-enriched Artemia had the highest percentage of eicosapentaenoic acid (EPA), 20:5(n−3). EPABSF/spirulina-enriched Artemia had the highest percentages of C18 fatty acids. Percentage of polyunsaturated fatty acids (PUFA) and n−3 PUFAs were highest in Super Selco®- and Algamac-3050-enriched Artemia. Proximate analysis revealed little difference among all four enrichment treatments in terms of protein, fat and carbohydrate levels.

The results from this experiment demonstrate that when culturing H. abdominalis, all three commercial enrichment products produce good seahorse growth and excellent survival. However, on a cost/benefit basis, all three commercial products were outperformed by the cheaper Artemia on-growing mixture (EPABSF/spirulina).     

Furunculosis in Atlantic salmon (Salmo salar L.) is not readily controllable by bacteriophage therapy

The potential of bacteriophage therapy to control bacterial disease in farmed fish was tested using, as an example, furunculosis of Atlantic salmon, caused by Aeromonas salmonicida subsp. salmonicida.

In vivo testing with Atlantic salmon and rainbow trout (Oncorhynchus mykiss Walbaum) showed no adverse effects, with bacteriophage generally cleared within 96 h of administration by either intraperitoneal (i.p.) injection or oral in-feed.

Juvenile Atlantic salmon were administered a combination of bacteriophage O, R and B (1.9 × 10⁸ pfu fish^− 1) by i.p. injection, after they had been challenged with A. salmonicida subsp. salmonicida 78027, also by i.p. injection. The fish that were injected with bacteriophage immediately after challenge died at a significantly slower rate then those that were either not treated with bacteriophage, or treated 24 h post-challenge. However, the end result (100% mortality) was not affected.

In further experiments the effects of oral (1.88 × 10⁵ pfu g^− 1 fish^− 1 daily for 30 days), bath (1.04 × 10⁵ ml^− 1 daily for 30 days) and i.p. (6.25 × 10⁷ pfu fish^− 1) phage treatment to control furunculosis in experimentally infected Atlantic salmon were compared with antibiotherapy (treatment with 10 mg kg^− 1 bw^− 1 day^− 1 oxolinic acid for 10 days), using an indirect cohabitation challenge. No protection was offered by any of the bacteriophage treatments, compared to the positive challenge group, although significant protection was offered by the oxolinic acid treatment. Analysis of samples taken from the trials demonstrated that bacteriophage were correctly administered to the fish and, on occasion, were isolated from fish that had succumbed to furunculosis. It was also shown that bacteriophage resistant A. salmonicida subsp. salmonicida isolates could be recovered from mortalities in all the treatment groups.

The results suggest that, although there were no safety problems associated with the approach, furunculosis in Atlantic salmon is not readily controllable by application of bacteriophage.     

Enrichment of Artemia with free methionine

Different enrichment procedures of the free amino acid (FAA) methionine were tested for Artemia nauplii. A direct enrichment protocol (methionine dissolved in the culture water) was compared with liposome enrichment protocols that varied in their membrane composition. During 16 h of direct enrichment in 5.3 mM methionine, the nauplii increased their content of free methionine between 20 and 30 times compared to the unenriched control (43.1±1.2 and 68.4±3.8 pmol·nauplius⁻¹ in two separate experiments vs. 2.4±1.0 pmol·nauplius⁻¹ in control). However, by encapsulating the identical amount of methionine into liposomes made from pure egg yolk phosphatidylcholine (PC) (>99% PC) and cholesterol, the nauplii content of free methionine reached 148.8±27.6 pmol·nauplius⁻¹, which is approximately 60 times more than in the unenriched control. Another liposome composition tested, made from crude egg yolk PC (>60% PC) and cholesterol, resulted in 90.5±4.1 pmol·nauplius⁻¹. The enriched nauplii still retained 80% of the free methionine after 8 h of incubation at conditions simulating feeding for Atlantic halibut larvae (13°C, 33.5 g·l⁻¹).

In conclusion: (1) Artemia nauplii can successfully be enriched with free methionine, (2) the high retention of free methionine in the Artemia nauplii following transfer to fish tanks shows that it is possible to offer fish larvae a feed with a high level of FAA, based on enrichment of Artemia nauplii.     

Streptococcus iniae infection and tissue distribution in hybrid striped bass (Morone chrysops×Morone saxatilis) following inoculation of the gills

This study was designed to test the possibility that Streptococcus iniae enters through the gills and causes infection in hybrid striped bass. To determine the dose response, four groups of fish were inoculated with S. iniae via the gills with a dose of 5.0×10⁵, 2.6×10⁶, 5.0×10⁶, or 1.0×10⁸ CFU/fish. One group of fish was inoculated with tryptic soy broth (TSB) via the gills to serve as controls. The cumulative percent mortality was 13%, 27%, 100% and 100% for 5.0×10⁵, 2.6×10⁶, 5.0×10⁶ and 1.0×10⁸ CFU/fish, respectively. We also examined the tissue dissemination of S. iniae at 0.5, 4, 8, 12, 24, 48 and 72 h after experimental gill inoculation. Fish were inoculated with 2.6×10⁶ or 5.0×10⁶ CFU/fish, which caused low and high mortality, respectively. Within 48 h, fish inoculated with the 2.6×10⁶ dose were culture positive on the gill surface, blood of the first and second gill arches, blood of the third and fourth gill arches and the nares. However, for the dose of 5.0×10⁶ CFU/fish, S. iniae was also isolated from the olfactory, optic and cerebellum regions of the brain, eye, head kidney, trunk kidney, spleen and liver at 48 h. For the 2.6×10⁶ dose, S. iniae was not isolated until 48 h post-inoculation, but was isolated at 12 h for the 5.0×10⁶ dose. The results of this study indicate that S. iniae can enter hybrid striped bass through the gills. However, mortality at similar S. iniae doses was lower than we previously observed by inoculation of the nares.     

Pharmacokinetics and the active metabolite of enrofloxacin in Chinese mitten-handed crab (Eriocheir sinensis)

The pharmacokinetics and active metabolite of enrofloxacin were estimated after single intramuscular administration (10.0 or 20.0 mg/kg body weight) to the Chinese mitten-handed crab (Eriocheir sinensis) in fresh water at 25.0 ± 1.0 °C. Levels of enrofloxacin and its metabolite ciprofloxacin in the main tissues (hemolymph, hepatopancreas, muscle, ovary and spermary) were simultaneously detected by HPLC. Enrofloxacin concentration–time profiles for the hemolymph in both tests were described by a two-compartment open model with first-order absorption. Distribution half-time (T_1/2), elimination half-time (T_1/2β), body clearance (CL/F), mean residence time (MRT_0–∞), area under the concentration–time curve from 0 to ∞ h (AUC_0–∞) and apparent volume of distribution (Vd/F), which derived from the pharmacokinetic model, were 0.427 h, 21.3 h, 0.133 l/h/kg, 60.0 h, 96.9 μg/ml/h and 4.08 l/kg, respectively, at a dose of 10.0 mg/kg body weight, and 0.216 h, 12.3 h, 0.189 l/h/kg, 85.8 h, 187 μg/ml/h and 3.35 l/kg, respectively, at a dose of 20.0 mg/kg body weight. Similarities were found between the hemolymph concentration–time curves of the two tests; for example, instant absorption process followed by the distribution phrase, and a second absorption peak at 6 h post-treatment. After intramuscular administration of 10.0 mg/kg body weight, absorption of enrofloxacin was observed in the main edible tissues (hepatopancreas, muscle, ovary and spermary), and the drug residue was the highest in the hepatopancreas, where the ‘drug sink’ phenomenon occurred. Comparative pharmacokinetics showed fast absorption, broad distribution and fast elimination of enrofloxacin in E. sinensis after intramuscular dosing. Regarding ciprofloxacin, the main active metabolite of enrofloxacin, though relatively low levels were detected in all the main tissues of the crab, its kinetics in the hemolymph in the two tests were not described by a one- or two-compartment open model.     

Effect of docosahexaenoic acid enrichment in Artemia on growth of Pacific bluefin tuna Thunnus orientalis larvae

Feeding enriched Artemia induces growth failure in Pacific bluefin tuna (PBT) Thunnus orientalis larvae; however, feeding of yolk-sac larvae of marine fish promotes larval growth. It is considered that this growth failure partly results from dietary docosahexaenoic acid (22:6n-3, DHA) deficiency. Therefore, we examined the effect of DHA contents in enriched Artemia on the growth of PBT larvae. Artemia nauplii were enriched with graded levels of DHA ethyl ester, and fed to PBT larvae for 9 days. Yolk-sac larvae of Japanese parrotfish Oplegnathus fasciatus were used as a reference diet. The DHA contents in Artemia increased with the enrichment from 0 mg g^− 1 dry weight basis (DW) to 25 mg g^− 1 DW, while the content in the reference diet was 21 mg g^− 1 DW. Feeding of enriched Artemia significantly improved the growth of PBT larvae. However, this improvement was negligible when compared with the excellent growth of the larvae that were fed the reference diet. PBT larvae that were fed the reference diet accumulated approximately twice or much higher levels of DHA in the neutral and polar lipids in the body when compared with the larvae that were fed enriched Artemia. These results show that PBT larval growth cannot be promoted by feeding enriched Artemia even if the DHA contents in Artemia are elevated to the same levels as those of yolk-sac larvae. The incorporation of dietary DHA into phospholipids in the fish body may be desirable for the normal growth of PBT larvae.     

Use of formulated diets as replacements for Artemia in the rearing of juvenile American lobsters (Homarus americanus)

American lobsters, Homarus americanus, have been successfully reared in hatchery operations for over a century, yet formulated diets have never been commercially produced. In recent years, commercial Artemia replacement (CAR) diets have been developed and marketed for use in aquaculture production of marine shrimp. Three separate experiments assessed the utility of rearing American lobsters on these shrimp CAR diets. First, survival and growth of stage IV American lobsters fed one of three CAR diets (Artemac 5, CAR1; Economac 4, CAR2; and Progression 3, CAR3) were compared to those of animals fed frozen adult n-3 fatty acid enriched Artemia. Survival to 3 months was highest for animals fed CAR3 (85%), while animals fed Artemia had the greatest weight gain (> 6 % day^− 1). A cost / benefit ratio analysis showed that CAR2 was the most cost efficient for juvenile production because of its low overall purchase cost. Second, stage IV lobsters were fed either CAR2 or frozen adult n-3 fatty acid enriched Artemia exclusively, or in combination (2 : 5, and 5 : 2). Again, CAR2 was a cost effective feed to use, even as a partial replacement for Artemia. Survival was higher in diets that included CAR2, and feeding it two days per week compensated for low quality Artemia. Finally, 1.5 year old lobsters fed a gelatin-bound mix of 80% CAR2 and 20% frozen Artemia for five months survived and grew equally well compared to lobsters fed gelatin-bound frozen adult Artemia, and better than a custom formulated maintenance diet. The benefits of incorporating formulated feeds into American lobster rearing programs to increase the effectiveness of enhancement programs is discussed.     

Hemagglutinating activity and electrophoretic pattern of hemolymph serum proteins of Penaeus monodon and Macrobrachium rosenbergii to white spot syndrome virus injections

The giant fresh water prawn Macrobrachium rosenbergii is known to be highly tolerant to white spot syndrome virus (WSSV) infections when compared to the widely cultured marine tiger shrimp Penaeus monodon. At present, the exact mechanism of tolerance by M. rosenbergii to WSSV is not known. In this study, we attempt to study the effect of WSSV injections on the hemagglutination activity of the hemolymph serum of both P. monodon and M. rosenbergii and look for changes if any, on their hemolymph serum protein electrophoretic patterns. Our results show that M. rosenbergii had significantly (p < 0.05) higher hemagglutinating activity against mouse erythrocytes when compared to P. monodon. As the infection progressed to 48 h there was a further increase (p < 0.05) in the hemagglutination activity in M. rosenbergii, while it decreased in P. monodon. 12% SDS-PAGE analysis of the hemolymph serum of M. rosenbergii infected with WSSV did not show any new protein bands, whereas few bands with decreased intensity was observed in moribund P. monodon where the hemagglutinating activity was also observed to be decreased. The results indicate that hemolymph hemagglutinin levels are modulated in crustaceans as a response to viral infections.     

Substitution of Chaetoceros muelleri by Spirulina platensis meal in diets for Litopenaeus schmitti larvae

The nutritional response of Litopenaeus schmitti larvae to substitution of Chaetoceros muelleri by Spirulina platensis meal (SPM) was evaluated. The substitution levels (S) were 0%, 25%, 50%, 75% and 100%, dry weight basis. Final larval length (FL) ranged from 1.98 to 3.16 mm for the different substitution levels. There was a significant relationship between S and FL, described by the following quadratic equation: FL = 2.853 + 0.01598S − 0.000233S². The substitution level (S) yielding maximum FL was 34.2%. Development index (DI) values ranged from 2.84 to 3.93 and were dependent on substitution level. The corresponding equation was DI = 3.799 + 0.00945S − 0.000189S² (P < 0.01). Maximum DI was obtained at 25.0% substitution. Survival was high (82–87%) and no significant differences were found between treatments. Protein digestibility of either microalgae was high, with 92% for SPM and 94% for C. muelleri, with no significant differences between them. The results in this study indicate that an adequate balance of nutrients in relation to the requirements of the species is critical. To simultaneously improve FL and DI, a 30% substitution of C. muelleri by SPM is suggested. This is equivalent to feeding 0.15 mg larvae^− 1 day^− 1 dry weight basis of a 70% C. muelleri/30% SPM diet, representing 0.078 mg protein larvae^− 1 day^− 1, 0.026 mg lipids larvae^− 1 day^− 1 and 2.732 J larvae^− 1 day^− 1.     

Dual disease resistance in a selectively bred eastern oyster, Crassostrea virginica, strain tested in Chesapeake Bay

Selective breeding efforts have yielded oyster strains, Crassostrea virginica, with improved survival and resistance against Haplosporidium nelsoni (MSX); however, because of susceptibility to the oyster pathogen Perkinsus marinus (Dermo), their utility has been limited in areas where the two parasites co-occur. Dual resistance to H. nelsoni and P. marinus was achieved through four generations of artificial selection of wild Delaware Bay oyster progeny at a site in the lower York River, Virginia, USA where both diseases are enzootic. During 1993–1995, survival, growth, and disease susceptibility of third generation Delaware Bay (F₃-DEBY) oysters were evaluated at the York River site in comparison to that of similarly selected third generation James River, Virginia oysters (F₃-JR), and first generation Louisiana oysters (F₁-LA), whose parents were naturally selected in the wild for resistance to P. marinus. During 1997–1999, the performance of F₄-DEBY was evaluated at three sites in Virginia in comparison to two groups of first generation oysters whose parents originated from Mobjack Bay and Tangier Sound, Virginia where both P. marinus and H. nelsoni are enzootic. In the presence of high infection pressure from both H. nelsoni and P. marinus, the F₃-DEBY stock showed significantly higher survival and growth than either the F₃-JR or F₁-LA strain. After 15 months of deployment, 79% of F₃-DEBY, 11% of F₃-JR and 17% of F₁-LA oysters were market size (≥76.2 mm) and cumulative mortality was only 16% in F₃-DEBY as compared to 42% in F₃-JR and F₁-LA. At the termination of the study, F₃-DEBY oysters exhibited 22% lower mortality than the F₁-LA stock, which outperformed the F₃-JR stock. Relative performance in respect to disease varied considerably with sample date; however, average H. nelsoni weighted prevalence varied such that F₃-DEBY1-LA=F₃-JR and average P. marinus weighted prevalence varied such that F₁-LA3-DEBY3-JR. In the 1997–1999 trial, F₄-DEBY oysters experienced 34–61% lower mortality, greater growth rate, and consistently lower prevalence and intensity of P. marinus than either Mobjack Bay or Tangier Sound oysters. H. nelsoni prevalences were very low (<12%) in all three stocks. This is the first study to demonstrate that reduced susceptibility to both P. marinus and H. nelsoni can be achieved through selective breeding.     

Immunological and pathological status of gilthead sea bream (Sparus aurata L.) under different long-term feeding regimes

The possible influence of the feeding regime (FR) on the immune system and pathological status of gilthead sea bream was studied. Two growth trials were performed starting at different seasons (trial 1=March; trial 2=June) under controlled experimental conditions. In both trials, FR-1 groups received a restricted amount of food, whereas FR-2 groups were fed to visual satiety. The pathology study included parasitological and bacteriological examination, and the immunological traits analysed were respiratory burst activity of head kidney leucocytes, serum lysozyme and alternative pathway complement activity (ACH₅₀). The immunological status of gilthead sea bream not only was not impaired by the restricted feeding regime, but also seemed to be enhanced in some aspects, as the respiratory burst of FR-1 fish of trial 2 was significantly higher. No differences in the bacterial isolates were detected between the two feeding regimes, and Vibrio harveyi was the most prevalent species in both cases, especially in warm months. Also, fish under FR-1 regimes had significantly lower mortality, lower prevalence of infection of all the parasites except Cryptosporidium molnari, and less histopathological alterations in liver and intestine than those under FR-2 regimes.     

Feeding has the largest effect on the ammonia excretion rate of the southern rock lobster, Jasus edwardsii, and the western rock lobster, Panulirus cygnus

The total ammonia nitrogen (TAN) excretion of spiny lobsters Jasus edwardsii and Panulirus cygnus, was determined in relation to temperature, body weight, emersion, daily rhythm and feeding. Temperature and body weight had large influences on the rate of TAN excretion. Exponential relationships were found between temperature (T) and TAN excretion of both species. These were described by the following equations: J. edwardsii Log₁₀ TAN=0.041T−3.57 (r²=0.979, F=143.2, P=0.001), P. cygnus Log₁₀ TAN=0.057T−3.90 (r²=0.987, F=302.2, P<0.001). TAN excretions of both species were positively correlated to body weight (W), and the relationships were described by the following equations: J. edwardsii Log₁₀ TAN=0.473 log₁₀ W−1.704 (r²=0.42, F=14.05, P=0.001), P. cygnus Log₁₀ TAN=0.499 log₁₀ W−1.346 (r²=0.69, F=44.18, P<0.001). TAN excretion increased significantly when lobsters were re-immersed after a 30 min period of emersion. However, it returned to pre-emersion levels by the second hour of re-immersion. Daily rhythm resulted in a significantly higher nocturnal TAN excretion rate for J. edwardsii; no daily rhythm was observed for P. cygnus. Feeding had the largest influence on TAN excretion, with maximum increases of 6.28 (J. edwardsii) and 5.60 (P. cygnus) times the pre-feeding level. TAN excretion rates remained significantly higher than the pre-feeding levels for an extended period (26 h, J. edwardsii; 30 h, P. cygnus). Implications for the use of purging tanks in lobster holding facilities and for the design of biofiltration systems are discussed.     

感染溶藻弧菌对日本蟳肝胰腺免疫功能的影响

摘 要：为探讨日本蟳感染溶藻弧菌的致病机理,采用溶藻弧菌人工感染健康日本蟳,分析了感染前后日本蟳肝胰腺 PO、SOD和LSZ活性变化及注射免疫多糖后的免疫保护率,并研究了细胞超微结构的病理组织学。结果表明：日本蟳感染溶藻弧菌后,肝胰腺中PO、SOD和 LSZ的活性均受到不同程度的影响,感染6~12 h,3种酶的活性均有一个上升的过程,但之后随时间的延长呈现下降趋势;免疫感染组因感染弧菌前注射了免疫多糖,日本蟳肝胰腺中PO、SOD和LSZ酶活性均显著高于感染组的酶活性,感染7 d后的免疫保护率高达72.73％,表明免疫多糖可提高酶的活性,使机体的抗病菌能力增强。超微结构显示：对照组日本蟳肝胰腺细胞结构形态正常,上皮细胞表面的微绒毛排列整齐,各细胞器结构完整;免疫组内质网、糖原颗粒和线粒体丰富,微绒毛排列致密;感染组肝胰腺组织受到明显的损伤,细胞和部分微绒毛脱落、受损;线粒体和粗面内质网变形、水肿或仅剩一空泡;核高度异染色质化,核膜破裂。与感染组相比,免疫后感染组具有形态结构较正常的细胞核、整齐致密的微绒毛和丰富的溶酶体,但依然可见水肿的内质网和狭长畸变的线粒体。     

Artemia enriched with high n-3 HUFA may give a large improvement in performance of Atlantic halibut (Hippoglossus hippoglossus L.) larvae

Atlantic halibut larvae were fed Artemia enriched with two different oil emulsions (cod liver oil and 2050TG) from first feeding to 70 days after first-feeding (dpff). Larvae fed 2050TG enriched Artemia had better growth, survival and eye migration than larvae fed the cod liver oil enriched Artemia, while pigmentation rate was similar in the two groups. In addition to the difference in fatty acids, the two emulsions differed in lipid class composition, since 2050TG is a synthetic oil and a mixture of mono-, di- and tri-acylglycerol, while cod liver oil is a tri-acylglycerol. Total lipid level, estimated as fatty acid methyl esters (FAME) was similar in the two Artemia types, but sum of n-6 and n-3 fatty acids, arachidonic acid (20:4n-6, ARA), docosahexaenoic acid (22:6n-3, DHA) and eicosapentaenoic acid (20:5n-3, EPA) were higher in Artemia enriched with 2050TG than in the cod liver oil enriched Artemia. However, the main difference in fatty acid composition in the larvae, was a higher DHA (% of total fatty acids) in 2050TG larvae than in cod liver oil larvae. The lipid level measured as FAME was up to four times higher in the 2050TG larvae than in the cod liver oil larvae, and the reason for this may have been a better bioavailability of the partly digested lipid in the 2050TG emulsion. The correlation between a high level of lipid in the larval tissues (e.g. high energy status) and improved eye migration in larvae fed the 2050TG enriched Artemia supports the hypothesis that energy limitation on the larval stage may be a cause of the impaired eye migration commonly observed in farmed Atlantic halibut juveniles.     

Loma salmonae (Protozoa: Microspora) infections in seawater reared coho salmon Oncorhynchus kisutch

Loma salmonae (Putz et al., 1965) infections were observed in five groups of coho salmon, Oncorhynchus kisutch, reared in seawater net-pens in Washington State, U.S.A. in 1984–1986. Ultrastructural characteristics, size of spores, tissues and host infected, and geographical location identified the microsporidium as Loma salmonae. Preserved spores measured 4.4×2.3 (4–5.6×2–2.4) μm and exhibited 14–17 turns of the polar filament. Infections were evident in the gills of some fish before seawater entry, but few parasites were observed and they caused little tissue damage. Infections observed in fish after transfer to seawater were associated with significant pathological changes in the gills. A mixed inflammatory infiltrate was associated with ruptured microsporidian xenomas within the vessels and interstitium of the primary lamellae. Microsporidian spores were dispersed throughout the lesions and were often seen inside phagocytes. The parasite was also observed in the heart, spleen, kidney and pseudobranchs; however, the inflammatory lesions were common only in the heart.

Monthly examination of fish after transfer to seawater showed peak prevalences (33–65%) of gill infections during the summer. Although moribund fish were often infected with other pathogens, the high prevalence of L. salmonae infections and the severity of the lesions it caused, suggested that this parasite significantly contributed to the recurrent summer mortalities observed at this net-pen site.     

The utilization of a filamentous green alga (Cladophora glomerata (L) Kutzin) as a protein source in pelleted feeds for Sarotherodon (Tilapia) niloticus fingerlings

Duplicate groups of S. niloticus fingerlings were fed each of five different diets for 8 weeks. The diets contained 30% crude protein supplied by varying proportions of fishmeal and C. glomerata meal. The five diets were formulated to supply fishmeal protein: C. glomerata meal protein ratios of 30 : 0 (diet 1), 25 : 5 (diet 2), 20 : 10 (diet 3), 15 : 5 (diet 4) and 10 : 20 (diet 5) respectively. A sixth diet (diet 6) containing 25% crude protein supplied entirely by C. glomerata meal was also fed.

Diets 1 and 2 produced significantly the highest growth rates and best protein utilization. Highest values for protein digestibility were found in diets 2 and 3 (94.5 and 93.9% respectively) although the values were not significantly different from other treatments. Weight gain and protein utilization decreased as the level of algal protein increased in diets 2 to 6. Carcass analysis revealed a decrease in lipid and protein contents with increasing levels of algae in the diet. It is postulated that algal meals are viable partial dietary protein sources for tilapia.     

Efficacy of formalin, hydrogen peroxide, and sodium chloride on fungal-infected rainbow trout eggs

Antifungal agents are essential for the maintenance of healthy stocks of fish and their eggs in intensive aquaculture operations. In the USA, formalin is the only fungicide approved for use in fish culture. However, hydrogen peroxide and sodium chloride have been granted low regulatory priority drug status by the United States Food and Drug Administration (FDA) and their use is allowed. We evaluated the efficacy of these fungicides for controlling fungal infections on rainbow trout eggs. A pilot study was conducted to determine the minimum water flow rate required to administer test chemicals accurately in Heath incubators. A minimum water flow rate of 7.6 1 min⁻¹ was necessary to maintain treatment concentrations during flow-through chemical exposures. The antifungal activity of formalin, hydrogen peroxide, and sodium chloride was evaluated by treating uninfected and 10% fungal-infected (Saprolegnia parasitica) rainbow trout eggs (Oncorhynchus mykiss) for 15 min every other day until hatch. There were no significant differences among treatments in percent hatch or final infection for uninfected eggs receiving prophylactic chemical treatments. Eggs of the negative control group (uninfected and untreated) had a mean hatch exceeding 86%. All chemical treatments conducted on the infected egg groups controlled the spread of fungus and improved hatching success compared with the positive control groups (infected and untreated). Formalin treatments of 1000 and 1500 μl l⁻¹ and hydrogen peroxide treatments of 500 and 1000 μl l⁻¹ were the most effective. Sodium chloride treatments of 30 000 mg l⁻¹ improved fry hatch, but the compound was less effective at inhibiting fungal growths compared with hydrogen peroxide and formalin treatments.