Efficacy of tricaine methanesulphonate and clove oil as anaesthetics for juvenile cobia Rachycentron canadum

Six experiments were designed to determine the optimal anaesthetic dosage of tricaine methanesulphonate (TMS) and clove oil that could be used safely on juvenile cobia Rachycentron canadum of two sizes [G1=4.9±0.8 g; G2=13.9±3.1 g]. We documented the stage of anaesthesia and the acute toxicity as 96 h LC₅₀ (lethal concentration 50% population) at various exposure times of the two anaesthetics. At 10 min induction time, the TMS 96 h LC₅₀ was 93.9 mg L⁻¹ in G1 and 97.0 mg L⁻¹ in G2. Compared with clove oil, the 96 h LC₅₀ was 60.0 mg L⁻¹ in G1 and 69.8 mg L⁻¹ in G2. The difference between the two groups (G1, G2) did not influence anaesthesia safety ( P >0.05). Rachycentron canadum achieved stage 3 anaesthesia more rapidly at a lower clove oil concentration level (40 mg L⁻¹, 10 min) than TMS (60 mg L⁻¹, 10 min), but the recovery period of clove oil, was significantly longer. Clove oil was the most effective in reducing the short-term stress induced by routine biometry (20 mg L⁻¹, 10 min) and also by transporting (1 mg L⁻¹, 8 h). Whereas, for long-term exposure, 40 mg L⁻¹ TMS was found to be safe.     

Olfactory nerve response of masu salmon (Oncorhynchus masou Brevoort) and rainbow trout (O. mykiss Walbaum) to clove oil and MS-222

Two anaesthetics, clove oil and methane sulphonate (MS-222), were examined for their effects on the olfactory nerve response of masu salmon ( Oncorhynchus masou Brevoort) and rainbow trout ( O. mykiss Walbaum). Exposing both species to clove oil for 3 min at concentrations of 50 and 100 mg L⁻¹, or for 10 min at 50 mg L⁻¹, did not significantly reduce their olfactory response. Directly applying clove oil anaesthesia to the olfactory epithelium significantly reduced olfactory response though after 20 min, olfactory response recovered to 70% and 52% of pre-treatment levels in masu salmon and rainbow trout respectively. Compared with the post-anaesthetic recovery of responses after clove oil (50 mg L⁻¹), buffered MS-222 (100 mg L⁻¹) with NaHCO₃ (100 mg L⁻¹), and unbuffered MS-222 (100 mg L⁻¹) treatment for 3 min, the response after MS-222 treatment declined gradually and significantly, but not after clove oil and MS-222+NaHCO₃ treatments. Clove oil appears to be an effective and relatively safe anaesthetic for salmonids with little long-term impact on their olfactory response, which plays a crucial role in their life history.     

Effect of combined photoperiod, water calcium concentration and pH on survival, growth, and moulting of juvenile crayfish (Procambarus clarkii ) cultured under laboratory conditions

The red swamp crayfish, Procambarus clarkii (Girard), is one of the most commonly farmed freshwater species in inland China due to its high market value and consumer demand. The aim of this study was to determine the optimum combinations of photoperiod, water calcium concentration and pH for juvenile survival, growth and moulting. In our orthogonal experiment, the three environmental factors were varied at three levels (photoperiod: 16L:8D, 12L:12D and 8L:16D; calcium concentration: 45.5, 65.5 and 85.5 mg L⁻¹; and pH: 6.8, 7.8 and 8.8). Range analysis showed that the maximum survival of juvenile crayfish occurred at photoperiods of 16L:8D or 8L:16D, water calcium concentration of 45.5 mg L⁻¹ and pH of 7.8; maximum weight gain at photoperiod 16L:8D, water calcium concentration 65.5 mg L⁻¹ and pH 7.8; maximum length increase at photoperiod 16L:8D, water calcium concentration 65.5 mg L⁻¹ and pH 7.8; and the highest moult frequency at photoperiod 12L:12D, water calcium concentration 65.5 mg L⁻¹ and pH 7.8. Analysis of variance indicated that photoperiod, water calcium concentration and pH significantly influenced only the weight gain of juvenile crayfish ( P <0.05). Taking growth into consideration, we suggest that a photoperiod of 16L:8D, calcium concentration of 65.5 mg L⁻¹ and pH 7.8 might be optimal conditions for rearing juvenile P. clarkii .     

Anaesthetic efficacy and physiological responses to clove oil-anaesthetized kelp grouper Epinephelus bruneus

The efficacy of clove oil as an anaesthetic and at producing a physiological response (plasma cortisol and glucose) was evaluated in the kelp grouper, Epinephelus bruneus . To acquire complete anaesthesia in less than 3 min and recovery in <10 min, three doses of clove oil were tested at 18, 22 and 26 °C. Although higher anaesthetic doses resulted in shorter induction times and longer recovery times, and a lower temperature resulted in longer anaesthesia induction and slower recovery, we found the optimal dose and administering temperature of clove oil to be 250–300 mg L⁻¹ at water temperature of 18 °C, 150–200 mg L⁻¹ at water temperature of 22 °C and 50–100 mg L⁻¹ at water temperature of 26 °C respectively. Following the administration of 150 mg L⁻¹ of clove oil at 22 °C, the plasma cortisol level was highest (4.24 ± 1.571 μg dL⁻¹) after 12 h and the plasma glucose was highest (92.7 ± 9.61 mg dL⁻¹) after 2 h. These results should be useful to the aquaculture industry, where anaesthesia is necessary for a range of activities.     

A method to determine protein digestibility of microdiets for larval and early juvenile fish

A method to evaluate protein quality using in vivo methods was developed for larval fish. FluoSpheres^® fluorescent microspheres (10 μm) were incorporated into two test diets, our standard zein microdiet (ZMD) and a microdiet with identical ingredients except for the replacement of high quality fish meal with the same product cooked for 24 h at 80 °C (ZMD-CF). Several trials were performed to design a reliable method to test digestibility using FluoSpheres^® as a marker. The developed in vivo technique was tested on 35 days posthatch (dph) larval Atlantic cod ( Gadus morhua L.) and two tropical fish species in the early juvenile stage. The method took into account loss of total protein to the faecal pellet and water column. Apparent digestibility of protein in larval cod fed ZMD was significantly higher than that of larvae fed ZMD-CF ( P  < 0.05). A growth study to validate differences between the two diets showed significant differences in growth and survival of larvae fed ZMD versus ZMD-CF ( P  < 0.05). Further validation of our results was indicated through the use of a pH-stat method using enzymes extracted from 35 dph larval cod guts. This novel technique will be advantageous for researchers to evaluate feed ingredients for larval marine fish and is adaptable to many different areas of larval fish nutrition.     

Comparison of the effects of four anaesthetics on biochemical blood profiles of perch

This study investigated the feasibility of using clove oil, 2-phenoxyethanol, or Propiscin as an alternative to tricaine methane sulphonate (MS 222) as a fish anaesthetic, particularly in regard to reducing fish stress. The biochemical blood profiles of perch Perca fluviatilis L. anaesthetized with either MS 222 (100 mg L⁻¹), clove oil (33 mg L⁻¹), 2-phenoxyethanol (0.40 mL L⁻¹) or Propiscin (1.0 mL L⁻¹), and a non-anaesthetized control group were compared. Biochemical profiles were determined from blood samples collected before treatment in controls. For each anaesthetic tested, fish were divided into two groups, one sampled immediately after 10-min anaesthesia and a second, sampled 24 h after 10-min anaesthesia. The values determined in the present study suggested that internal organs and tissues of perch were slightly altered by MS 222, clove oil and 2-phenoxyethanol anaesthesia, but not by Propiscin anaesthesia.     

Nutrient values of dietary ascorbic acid (l-ascorbyl-2-polyphosphate) on growth, survival and stress tolerance of larval shrimp, Litopenaeus vannamei

l -ascorbyl-2-polyphosphate (ApP) was used as a vitamin C source to investigate the ascorbic acid (AsA) requirements on growth performance and stress resistance of the larval white shrimp, Litopenaeus vannamei . Five isoenergetic and isonitrogenous fish meal-fish protein hydrolysate-based diets with five levels of ApP, AsA equivalent to 91.8, 188, 271, 360 and 436 mg kg⁻¹ diet were fed to triplicate groups of L. vannamei (mean initial wet weight 1 mg) for 32 days. The diet with AsA 91.8 mg kg⁻¹ showed high cumulative mortality after 10 days of feeding. After the 32-day trial, the shrimp that fed the diet had significantly lower survival and weight gain (WG, %) than those that fed 188, 271, 360 and 436 mg AsA kg⁻¹ diets. Specific growth rate (SGR, % day⁻¹) and final body wet weight (FBW, mg) showed the same pattern as WG (%). There were no significant differences in growth performance (FBW, WG and SGR) among the groups that fed 188, 271, 360 and 436 mg kg⁻¹ of AsA at the termination of feeding trial. Broken-line regression analysis on WG indicated that 191 mg AsA kg⁻¹ in the diet was the optimum for larval L. vannamei . On the contrary, dietary level of more than 360 mg AsA kg⁻¹ was needed to ensure high resistance to stressful conditions such as low dissolved oxygen stressors.     

Bioencapsulation of erythromycin using adult brine shrimp, Artemia franciscana (Latreille)

Live adult brine shrimp, Artemia franciscana (Latreille), were enriched with erythromycin to determine if Artemia could accumulate therapeutic levels for subsequent feeding to young fish. Three trials were conducted to determine the erythromycin incorporation and survival rates of enriched Artemia when fed either liposomes containing erythromycin or various erythromycin suspensions. Erythromycin concentration in Artemia fed a liposome suspension was low (∼ 5 μg mL⁻¹) relative to Artemia fed the direct suspension (> 100 μg mL⁻¹) over the same time period. When enriched with suspensions up to 1 g erythromycin L⁻¹ sea water for 14 h, Artemia survival was not significantly affected ( P > 0.05) relative to controls. Using a suspension of 1 g L⁻¹, tissue erythromycin concentrations of 109 ± 16 μg erythromycin mL⁻¹  Artemia homogenate (mean ± SEM) were achieved after 12 h. Concentrations above 170 μg mL⁻¹ were obtained using suspensions of 2–5 g L⁻¹, but Artemia survival significantly ( P < 0.05) decreased.     

Use of salt to control ichthyophthiriosis and prevent saprolegniosis in silver perch, Bidyanus bidyanus

The ciliate protozoan, Ichthyophthirius multifiliis , and the fungus, Saprolegnia parasitica , cause the diseases ichthyophthiriosis and saprolegniosis respectively. Both diseases are difficult to control and can cause high mortalities of freshwater fish, including the Australian silver perch, Bidyanus bidyanus (Mitchell). The efficacy of salt (NaCl) in controlling and preventing these diseases in silver perch was evaluated in aquaria and tanks. Low pH levels were also evaluated as a control for ichthyophthiriosis. Concentrations of 2 or 3 g L⁻¹ salt controlled infestations of I. multifiliis , and fish were free of both theronts and trophonts by day 8 at temperatures of 17.3–21.3 °C and by day 6 at 19.2–23.5 °C. Fish treated with 1 g L⁻¹ salt remained infested and all fish in a control treatment (0 g L⁻¹ salt) died. Although the mean survival rates of infested fish at pH levels of 5 or 6 were only 13.9% and 7.6%, respectively, there were no theronts or trophonts on surviving fish after 12 days. Silver perch harvested from a pond and treated with 2 or 3 g L⁻¹ salt did not become infected with S. parasitica and survival was 100%, whereas 16.6% of untreated (0 g L⁻¹ salt) fish became infected and survival was only 66.7%. A concentration of 2 g L⁻¹ NaCl is recommended for the control of ichthyophthiriosis and the prevention of saprolegniosis in silver perch held in tanks, aquaria and re-circulating aquaculture systems.     

Continuous exposure to infectious pancreatic necrosis virus during early life stages of rainbow trout, Oncorhynchus mykiss (Walbaum)

Rainbow trout, Oncorhynchus mykiss (Walbaum), were exposed continuously to infectious pancreatic necrosis virus (IPNV) at 0, 10¹, 10³ or 10⁵ plaque forming units (pfu) L⁻¹ of water to estimate the effects of chronic IPNV exposure on early life stages. Fish density averaged 35 fish L⁻¹ (low density) or 140 fish L⁻¹ (high density), and the tank flow rate was 250 mL⁻¹ min. Virus exposure began at 6 days before hatch and continued until fish were 44 days old. Cumulative per cent mortality, analysis of survival and hazard functions, and discrete-time event analysis were used to explore the patterns of survival and mortality. In eggs and fish exposed to IPNV, mortality significantly greater than in the 0 pfu L⁻¹ exposure did not occur until IPNV concentration was 10⁵ pfu L⁻¹ at low fish density and 10³ pfu IPNV L⁻¹ at high fish density. These results suggest that in the natural aquatic environment, where rainbow trout densities are likely to be considerably lower than in this study, mortality resulting from infection with IPNV will very likely not occur when ambient concentrations of virus are ≤10³ pfu IPNV L⁻¹. In aquaculture rearing units, trout density is likely to be as high or higher than the densities used in this study. Therefore, continuous inputs of virus at concentrations greater than 10¹ pfu L⁻¹ may result in IPN epidemics in aquaculture facilities.     

The potential of liposomes as a nutrient supplement in first-feeding marine fish larvae

The ingestion rate (ng liposome larva^–1 h^–1) of extruded [1–¹⁴C] palmitic acid-labelled liposomes containing physiological saline (PHS) or cod fish extract (CFE), was tested in 5-day-old gilthead seabream Sparus aurata and white grouper Epinephelus aeneus larvae. A follow-up study compared the assimilation of radioactive free fatty acid (FFA) label of these two liposome treatments into six phospholipid and neutral lipid fractions as well as the nonlipid fraction in 5-day-old seabream. In seabream larvae, there was a 50-fold ( P  < 0.05) increase in the net consumption rate when fed CFE liposomes (2305.8 ng liposome larva^–1 h^–1) compared with liposomes containing physiological saline (42.7 ng liposome larva^–1 h^–1). A similarly significant ( P  < 0.05) but less marked pattern was also observed in the grouper larvae where the CFE treatment larvae ingested 238.5 ng liposome larva^–1 h^–1 compared with 54.3 ng liposome larva^–1 h^–1 in larvae fed the PHS liposomes. In seabream larvae ingesting CFE and PHS liposomes, radioactivity was found in all larval fractions analysed. However, marked treatment differences ( P  > 0.05) in assimilation were found only in the triacylglycerol fraction (3.4 and 0.6 dpm larva^–1 h^–1, respectively) and nonlipid fraction (11.2 and 15 dpm larva^–1 h^–1, respectively).     

Comparative efficacy of MS-222 and benzocaine as anaesthetics under simulated transport conditions of a tropical ornamental fish Puntius filamentosus (Valenciennes)

There is a growing commercial interest in the fish, Puntius filamentosus , in the ornamental fish trade in India and elsewhere. The trade is, however, hampered by severe mortalities during transport of the fish owing to insufficient data available on the use of anaesthetics. To resolve this problem, we evaluated the efficacy of two anaesthetics, MS-222 and benzocaine, in sedating P. filamentosus in simulated transportation experiments and used stress response parameters such as cortisol and blood glucose levels to perform assessments. We observed that MS-222 at 40 mg L⁻¹ and benzocaine at 20 mg L⁻¹ were sufficient to induce sedation for 48 h. Above these concentrations, both the anaesthetics adversely affected the fish and resulted in mortalities. Both anaesthetics significantly lowered the blood cortisol and glucose levels compared with the unsedated controls. Importantly, the anaesthetics treatment significantly lowered the post-transport mortality in the fish. The results of the study show that MS-222 and benzocaine could be used as sedatives to alleviate transport-related stress in P. filamentosus to improve their post-transport survival and hence reduce economic loss.     

Size-related changes in habitat selection by larval grayling (Thymallus thymallus L.)

Abstract  – In a medium-sized river in northern Finland, larval grayling shifted with growth from shallow habitats with slow velocities, fine substrata and abundant vegetation cover to deeper sites with swifter velocities, coarse substrata and sparse vegetation cover within 3 weeks. Small (17–21 mm) larvae preferred water depths 10–30 cm, substrata dominated by mud or sand (<2 mm), 10–70% vegetation cover and water velocities <10 cm · s⁻¹. Middle-sized (22–25 mm) larvae preferred 30–90 cm depths, sandy substrata, <40% vegetation cover and velocities <10 cm · s⁻¹. Large (26–31 mm) larvae preferred >50 cm depths, substrata dominated by sand or boulders, <20% vegetation cover and 10–50 cm · s⁻¹ velocities. The strict habitat requirements of the smallest larval group suggest that these habitats, in particular, are important to the early survival of grayling.     

Pharmacokinetics of oxolinic acid in gilthead sea bream, Sparus aurata L.

This is the first study on the pharmacokinetic parameters of oxolinic acid (OA) in gilthead sea bream, Sparus aurata L. The kinetic profile of OA was studied after a single intravascular injection (20 mg kg⁻¹) in 100 g fish at 20 °C. The distribution half-life ( t _1/2α) and the elimination half-life ( t _1/2β) of the drug were found to be short (0.51 and 12.60 h, respectively). The drug penetration from the plasma to the tissues was adequate as the apparent volume of distribution of the drug at steady-state ( V _d(ss)) was found to be 2.11 L kg⁻¹. The mean residence time ( MRT ) of OA was short (14.25 h) and the total clearance rate ( Cl _T) of the drug was low (0.15 L kg⁻¹ h⁻¹). The bioavailability ( F %) of OA following oral administration (30 mg kg⁻¹) was also low (14%). Maximum values were observed for muscle at 0.5 h after injection, with levels declining as with subsequent sampling. At the first two time points (0.5 and 1 h) plasma levels of OA were higher than muscle, however, the reverse was evident for subsequent samples. Following oral administration, highest muscle levels were found at 16 h and, with the exception of the 24-h sampling, muscle OA concentrations were higher than plasma at all time points. The fast elimination of OA suggests short withdrawal times with reference to human consumption of treated fish.     

Feeding selectivity of the marine ascidian Styela clava

The sessile suspension feeding ascidian, Styela clava , plays an important role in the functioning of estuarine systems. The feeding selectivity of S. clava was examined using standard physiological techniques in terms of particle sizes and diet concentrations. Styela clava increased its retention efficiency (the proportion of available particles retained in ascidian) with increasing particle size at a lower diet concentration [expressed as particulate organic matter (POM)] from 2.11 to 10.06 mg POM L⁻¹, and the highest retention was reached for 5–20 μm particles. With increasing diet concentration to 20.12 mg POM L⁻¹, S. clava retained small particles (<3.5 μm) more efficiently and large ones (>15 μm) less efficiently. The selectivity of ascidian with different sizes was similar at the same concentration. At every diet concentration, larger ascidians retained more 5–20 μm cells than the smaller ones. All these results indicate that S. clava possesses a particle selection mechanism that may affect the zooplankton communities to a certain degree.     

Growth and survival of larval and juvenile cobia Rachycentron canadum in a recirculating raceway system

Cobia Rachycentron canadum is a fast-growing, pelagic marine species that has recently attracted aquaculturists in both the research and commercial sectors. The typical method of grow-out for this species is in outdoor systems where production is limited to locations and seasons conducive for adequate growth and survival. Expanding the culture of cobia to indoor recirculating aquaculture systems (RAS) would allow for the production of fingerlings throughout the year and extend production to cooler regions. Two rearing trials were conducted to examine the growth and survival of cobia from hatching through 4 (trial 1, T1) or 35 (trial 2, T2) g in RAS. Cobia larvae were reared in circular tanks placed in a raceway to control water temperature and quality. During early juvenile grow-out, fish were transferred without grading to a second raceway on 29 dph (T1) or over a period of grading from 29–43 dph (T2). Larval growth (1–22 dph) measured as standard length was similar for both trials ranging from  3.9 to 14.7 mm. However, larval growth measured as wet weight (0.033 g, T1; 0.026 g, T2) or dry weight (5.7 mg, T1; 3.9 mg, T2) was significantly greater on 22 dph during T1 as was the ratio between myotome height and standard length. These differences may have resulted from an increase in initial densities from 8.7 larvae l^− 1 (T1) to 14.7 larvae l^− 1 (T2) which apparently caused an increase in food competition and overall aggression. During juvenile grow-out, cobia reached 4.0 g on 43 dph in T1 and 35.4 g on 71 dph in T2 matching weights achieved during grow-out in outdoor ponds. Over the course of both trials, survival was similar to that reported in outdoor ponds. Mean survival (± S.D.) during the early rearing phase (hatching through 29 or 43 dph) averaged 13.2 ± 3.2 % and 10.4 ± 3.2 % corresponding to final densities of 0.9 ± 0.2 and 1.2 ± 0.4 fish/l for T1 and T2, respectively. During the first grow-out phase (29–43 dph), survival of fish moved into the open raceway was 64.5% in T1 and 88.7 % in T2. Survival of cobia during the second grow-out phase (43–71 dph) for T2 was 92.5%. The results of this study indicate that cobia can be successfully cultured in indoor systems from hatching through at least 35 g without negatively affecting growth or survival.     

Vertebral deformity in cultured Atlantic cod larvae: ontogeny and effects on mortality

The effects of different egg incubation densities on the incidences of vertebral deformities in Atlantic cod larvae were investigated. Cod eggs were incubated at four different densities, 3, 6, 12 and 48 mL eggs L⁻¹, of water.  When all the eggs hatched, larvae were reared in 30 L glass aquaria. Larval samples were taken at 0, 14, 42 and 56 days post hatch (dph) for deformity analysis. Larval samples were stained using bone and cartilage staining methods to determine vertebral deformity. Incubation densities did not have any significant effects on vertebral deformities in Atlantic cod larvae. However, the incidence of larval vertebral deformity was high at hatch and decreased as the larvae grew older until 42 dph, indicating selective mortality of deformed larvae during this period. Larvae at 56 dph, however, showed an increase in the incidence of vertebral deformity, indicating a possible nutritional or prey-type effect. To our knowledge, no studies have documented the occurrence of variable patterns in vertebral deformities in cod at various developmental stages. Overall, our results suggest that broodstock husbandry, genetics and/or nutrition could play a major role in causing vertebral deformities in Atlantic cod at hatch; however, nutrition and prey type may play a major role during metamorphosis.     

Comparative growth performances of diploid and triploid Chinese shrimp Fenneropenaeus chinensis (Osbeck, 1765) under different salinities

This study aimed at evaluating the ploidy effects on growth performances of Chinese shrimp ( Fenneropenaeus chinensis Osbeck, 1765) reared in different salinities under laboratory conditions. In the acute salinity experiment, there was no difference ( P >0.05) in tolerance observed in triploid and diploid shrimp due to abrupt salinity changes. The lethal salinity for 50% of the individuals in 96 h at 23–25 °C was about 2 g L⁻¹ in both triploids and diploids. While for the chronic salinity experiment, statistical analyses confirmed that the differences in growth performances including the specific growth rate (SGR), the feeding rate (FR), feed conversion efficiency (FCE) and intermoult period (IP) between triploid and diploid were related to salinity. Diploid shrimp reared in 20 g L⁻¹ exhibited highest SGR ( P <0.05), while triploids performed well in 20 and 30 g L⁻¹ salinities ( P <0.05). Based on the survival and growth data, the optimal salinity for the culture of diploid F. chinensis should be 20 g L⁻¹ and for triploids it should be between 20 and 30 g L⁻¹.     

Evaluation of lipid spray beads for the delivery of water-soluble materials to a marine suspension-feeder, the Manila clam Tapes philippinarum (Deshayes 1853)

We describe the development and evaluation of a new microparticle for delivering low-molecular weight, water-soluble materials to suspension feeders. Spray beads successfully incorporated materials dissolved in an aqueous phase or as dry particulate, within a triacylglyceride bead composed of tripalmitin, 600 mg g⁻¹ tripalmitin/400 mg g⁻¹ triolein, or 600 mg g⁻¹ tripalmitin/400 mg g⁻¹ fish oil.
Riboflavin was successfully incorporated (up to 44 mg g⁻¹ lipid) and retained (up to 98% over 24 h in seawater) as dry particles in all three mixtures of lipid. Aqueous oxytetracycline hydrochloride or polymeric dye were incorporated (45.6 mg g⁻¹ lipid and 18.1 mg g⁻¹ lipid, respectively) and retained best (99% and 94%, respectively) in spray beads composed of tripalmitin. The addition of triolein or fish oil to the lipid bead reduced incorporation and retention efficiencies for aqueous core materials by up to 75%.
Manila clam seed readily ingested and digested lipid microparticles, spray beads and lipid-walled microcapsules. Microparticles composed of tripalmitin were excreted with their payloads intact. Intact microparticles composed of 600 mg g⁻¹ tripalmitin/400 mg g⁻¹ fish oil were largely absent in faecal strands suggesting successful release and delivery of microparticle contents to clams.
Spray beads composed of tripalmitin softened with 400 mg g⁻¹ fish oil represent an effective microparticle type for delivering low-molecular weight, water-soluble materials to aquatic suspension feeders.     

Effect of plant active compounds on immune response and disease resistance in Cirrhina mrigala infected with fungal fish pathogen, Aphanomyces invadans

The antibacterial activity of individual and mixed medicinal plant compounds, azadirachtin (Az), camphor (Ca) and curcumin (Cu), was tested at 100, 200, 300, 400, 500, 600 and 700 ppm (mg L⁻¹) against fungal fish pathogen, Aphanomyces invadans, in vitro . At the lower concentrations between 100 and 300 ppm, the mixture of the tri-herbal (Az+Ca+Cu) compound yielded a higher ( P >0.05) zone of inhibition (ZI) of 7 mm than the positive control; the maximum ZI values (8–15 mm) were realized between 400 and 700 ppm ( P <0.05). At the lowest minimum inhibitory concentration (MIC) the tri-herbal compound (100 ppm) yielded 13 colony-forming units; hence, this dose can be effectively used at the lowest concentration of 100 mg L⁻¹ to ward off the growth of A. invadans in vitro . In Cirrhina mrigala , intramuscular administration (100 μL) of the selected doses of 100, 400 and 700 ppm (mg L⁻¹) significantly enhanced ( P <0.05) the serum lysozyme activity (Ly), production of reactive oxygen species and reactive nitrogen species (RNS or NO) by peripheral blood leucocytes on the 10th, 20th and 30th day. A priori administration of the compound in the fish (100 ppm on 30th day) decreased the percentage mortality when challenged with the pathogen while in the untreated group the mortality increased ( P <0.05). This study indicates that intramuscular administration of the tri-herbal compound Az+Ca+Cu at a concentration of 100 ppm could augment the immune response in C. mrigala against A. invadans .