In vitro and in vivo antifungal activity of Satureja cuneifolia ten essential oil on Saprolegnia parasitica strains isolated from rainbow trout (Oncorhynchus mykiss,Walbaum) eggs

In the present study, the chemical composition and the antifungal properties against Saprolegnia parasitica (in vitro and in vivo) of the essential oils of thyme (Satureja cuneifolia) from Mediterranean region of Turkey were evaluated for the first time. The composition of oils was analysed using gas chromatography/mass spectrometry (GC/MS). The major constituents of oil of S. cuneifolia were cavracrol (46,84%) and cymene (16.90%). Antifungal effects of S. cuneifolia essential oil against S. parasitica strains (A1 and E1) were detected by disc diffusion and tube dilution assays. The antifungal effect of S. cuneifolia was determined to be stronger against S. parasitica E1 isolate (MIC 50 μL mL^?1, MLC 250 μL mL^?1) compared with S. parasitica A1 isolate (MIC 50 μL mL^?1, MLC 500 μL mL^?1). Following in vitro assays, effective doses of S. cuneifolia for disease control in rainbow trout eggs experimentally infected with S. parasitica were investigated. For this aim, infected eggs were treated with the essential oil (0, 5, 10, 20 and 50 ppm) during incubation period (21 days) after fertilization. Formalin (5 mL L^?1) was used as positive control. Hatching rate of eggs at the end of incubation period were calculated. The highest hatching rates were recorded in S. parasitica E1 strain at 5 and 10 ppm concentrations of S. cuneifolia and in S. parasitica A1 strain at 10 and 20 ppm (P < 0.05).     

Effect of oregano (Origanum onites L.) essential oil on growth,lysozyme and antioxidant activity and resistance against Lactococcus garvieae in rainbow trout,Oncorhynchus mykiss (Walbaum)

The aim of this study was to investigate the effects of different levels of Origanum onites L. essential oil as feed additives on the growth performance, antioxidant activity and disease resistance of rainbow trout. Fish (26.05 ± 0.15 g) were fed the experimental diets supplemented with four different concentrations (0.125, 1.5, 2.5 and 3.0 mL kg^?1) of O. onites essential oil for 90 days. Fish fed diets containing essential oil of O. onites had significantly higher final weight than the control group. Feed conversion ratio in fish fed diets containing 1.5 and 3.0 mL kg^?1 essential oil of O. onites was improved than other treatments (P < 0.05). The lowest feed conversion efficiency ratio was recorded in the 0.125 mL kg^?1 group of O. onites. Antioxidant status of fish was assayed for levels of plasma superoxide dismutase (SOD) and plasma catalase (CAT) activity. Lysozyme activity in plasma was significantly higher in fish fed diet containing 3.0 mL kg^?1 essential oil of O. onites (P < 0.05). After 8 weeks of feeding, fish were challenged with Lactococcus garvieae and cumulative mortality was recorded over 15 days. Dietary administration of 0.125, 1.5 and 2.5 mL kg^?1 O. onites significantly reduced fish mortality (P < 0.05). The 3.0 mL kg^?1 diet showed no mortality after challenged with L. garvieae. These results suggested that the essential oil of O. onites could be applied as growth promoter and also improved disease resistance when added to rainbow trout feed.     

Use of essential oils extracted from three Origanum species for disinfection of cultured rotifers (Brachionus plicatilis)

Essential oils extracted from three plant species of the genus Origanum, Origanum vulgare subsp. hirtum, Origanum onites L. and Origanum marjorana L., were assayed as potential antibacterial agents for disinfection of rotifers Brachionus plicatilis. Survival of rotifers in seawater with added oregano essential oils was significantly higher than in the control treatment (P < 0.05). When rotifers were incubated in seawater with added oregano essential oils at a final concentration of 10 mg L^?1 for 4 h, the numbers of total culturable bacteria were not significantly lower than in the control treatment (P > 0.05), while the numbers of presumptive Vibrio in rotifers were significantly reduced compared with the control treatment (P < 0.05).     

Chemical composition of Lippia spp. essential oil and antimicrobial activity against Aeromonas hydrophila

Phytotherapy can replace antibiotic administration as an alternative to control Aeromonas hydrophila, one of the main bacteria involved in the aetiology of farmed fish diseases. Given that plants of the Lippia spp. genus show biological potential for antimicrobial activity, this study evaluated the chemical composition of essential oils extracted from Lippia alba, Lippia origanoides and Lippia sidoides and their activity against A. hydrophila. The oils were obtained by steam distillation in a Clevenger‐type apparatus and their composition determined by gas chromatography and mass spectrometry (CG/MS). Antibacterial activity was assessed by calculating the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using broth microdilution method. The main compounds identified were geranial (25.4%) and neral (16.6%) in L. alba oil, carvacrol (40.4%) and p‐cymene (11.4%) in L. origanoides oil and thymol (76.6%) and ortho‐cymene (6.3%) in L. sidoides oil. The three Lippia species showed bacteriostatic and bactericidal action against A. hydrophila, with MICs and MBCs ranging from 1250 to 5000 μg mL^?1. Of the species tested, the best performance was obtained with essential oil of L. sidoides.     

Essential oils to control ichthyophthiriasis in pacu,Piaractus mesopotamicus (Holmberg): special emphasis on treatment with Melaleuca alternifolia

In vitro effect of the Melaleuca alternifolia, Lavandula angustifolia and Mentha piperita essential oils (EOs) against Ichthyophthirius multifiliis and in vivo effect of M. alternifolia for treating ichthyophthiriasis in one of the most important South American fish, Piaractus mesopotamicus (Holmberg), were evaluated. The in vitro test consisted of three EOs, each at concentrations of 57 μL L^?1, 114 μL L ^?1, 227 μL L^?1 and 455 μL L ^?1, which were assessed once an hour for 4 h in microtitre plates (96 wells). The in vitro results demonstrated that all tested EOs showed a cytotoxic effect against I. multifiliis compared to control groups (P < 0.05). The in vivo treatment for white spot disease was performed in a bath for 2 h day^?1 for 5 days using the M. alternifolia EO (50 μL L ^?1). In this study, 53.33% of the fish severely infected by I. multifiliis survived after the treatment with M. alternifolia (50 μL L ^?1) and the parasitological analysis has shown an efficacy of nearly 100% in the skin and gills, while all the fish in the control group died. Furthermore, the potential positive effect of M. alternifolia EO against two emergent opportunistic bacteria in South America Edwardsiella tarda and Citrobacter freundii was discussed.     

Prevention of Saprolegniasis in rainbow trout (Oncorhynchus mykiss) eggs using oregano (Origanum onites) and laurel (Laurus nobilis) essential oils

The present study investigated the antifungal effects of essential oils of oregano (Origanum onites) and laurel (Laurus nobilis) on Saprolegniasis, a disease that occurs in rainbow trout eggs during the incubation period. Oregano and laurel were ground after drying, and essential oils were obtained by water distillation method using a Clevenger apparatus. The essential oils were added to potato dextrose agar (PDA) at the rates of 1–1000 ppm, and the minimum inhibitory concentration (MIC) was determined as 250 ppm whereas the minimum lethal concentration (MLC) was determined to be 500 ppm for both plants. In the in vivo trials, fertilized eggs were treated with predetermined doses either by bathing during water hardening and incubation period or only during incubation period, and death rates were monitored during embryological development. The best larvae hatching rate was determined in 500 ppm oregano and 500 ppm laurel groups treated during water hardening plus daily as 82.11% and 79.87%, respectively. According to the results, it was determined that oregano and laurel essential oils exhibited better results in all doses compared with the negative control group, and 500 ppm dose had a better effect than the positive control group treated with formalin.     

Antifungal effect of Origanum onites essential oil as an alternative to formalin in the artificial incubation of narrow‐clawed crayfish (Astacus leptodactylus Eschscholtz, 1823)

As alternative to formalin, the antifungal effect of a plant product [Origanum onites L. (Lamiaceae) oil] was investigated for use in the artificial incubation of narrow‐clawed crayfish (Astacus leptodactylus Eschscholtz) eggs. For this purpose, this study was conducted as two experiments. In experiment I, the eggs were artificially incubated for 40 days. In experiment II, juveniles were cultured to determine effects of O. onites oil on juveniles for 30 days. The experimental groups were as follows: formalin (3500 ppm for 15 min), O. onites oil (300 ppm for 15 min, 700 ppm for 2 min and 1000 ppm as a dip treatment 15 split‐second) and a control (no treatment). In the experiment I, the highest hatching rate (86%) and survival rate of stage II juveniles (80%) were observed in 1000 ppm dip group. These results were similar to that of formalin group (85% and 79%) respectively. The control group exhibited the lowest hatching rate (49%) and stage II rate (42%) compared with the 1000 ppm dip group and 3500 ppm formalin treatments. However, other concentrations (300 and 700 ppm) of O. onites showed toxic effects on the eggs and there was no hatching. In the experiment II, the survival rate and growth performance of the crayfish juveniles were similar in all groups. This study indicated that the 1000 ppm O. onites dip treatment could be a good alternative to formalin for improved egg hatchability in the artificial incubation of crayfish eggs.     

In vitro antibacterial activity of four nano‐encapsulated herbal essential oils against three bacterial fish pathogens

Bacterial pathogens causing diseases in fishes have broad host ranges, and may cause high mortality or ongoing chronic infections. Pathogen treatments using antibiotics are becoming limited due to increasing concerns for the development of resistance and environmental dissemination of bacteria harboring resistance genes. This study aimed to determine whether nano‐emulsions of selected plant‐derived essential oils (Origanum vulgare, Eucalyptus globulus, Melaleuca alternifolia and Lavendula angustifolia) were bacteriostatic or bactericidal to Aeromonas hydrophila, Streptococcus iniae and Photobacteriumdamselae subspecies damselae. All treatments showed antibacterial activity, and in almost all cases the activity of the nano‐emulsions was superior to their essential oil counterparts. Origanum vulgare (oregano) nano‐emulsion had the most effective antibacterial activity, with a minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of 3.12 µg/ml against all three species; substantially better than tetracycline. The present study highlights the potential of new formulations of essential oils to enhance bacterial disease control in cultured fish.     

Effects of flow rate on growth performance and welfare of juvenile turbot (Scophthalmus maximus L.) in recirculating aquaculture systems

The effects of flow rate on growth and welfare of juvenile turbot (Scophthalmus maximus L.) were investigated in the present study. Fish with same initial weight (102.5 ± 10.6 g) were subjected to four flow rates, equalling to 0.5, 1, 1.5 and 2 tank volumes per hour in twelve 392 L tanks during 80 days. Results showed that specific growth rate of turbots increased (0.40–0.58% day^?1) significantly with promoted flow rate (P < 0.05). Total ammonia nitrogen, nitrite nitrogen, unionized ammonia nitrogen, chemical oxygen demand, total bacteria and total vibrio in tanks were affected significantly by flow rate and accumulations were found in low rate (200 L h^?1) (P < 0.05). Free carbon dioxide increased significantly with the increased flow rate and ranged between 4.5 and 13.5 mg L^?1 (P < 0.05). Both superoxide dismutase activity and lysozyme activity increased significantly with flow rate (P < 0.05), with ranges of 108.51–131.57 U mL^?1 and 551.81–869.28 U mL^?1. Serum cortisol showed reversed tendency and ranged between 7.39–19.26 ng mL^?1. The principal components analysis suggested that increased flow rate promoted fish welfare. It was concluded that increased flow rate promoted the growth of juvenile turbot, possibly explained by fish welfare differences in combination of health, water quality and serum parameters.     

The anaesthetic effect of camphor (Cinnamomum camphora), clove (Syzygium aromaticum) and mint (Mentha arvensis) essential oils on clown anemonefish,Amphiprion ocellaris (Cuvier 1830)

The aim of this study was to evaluate the use of clove (Syzygium aromaticum), camphor (Cinnamomum camphora) and mint (Mentha arvensis) essential oils as anaesthetics during the management of clown anemonefish (Amphiprion ocellaris). For 15 min, the animals were subjected to concentrations of 5, 10, 20, 27 and 35 μL L^?1 of clove oil, 17, 35, 50, 70 and 100 μL L^?1 of mint oil, and 200, 400, 500, 550 and 600 μL L^?1 of camphor oil (tested in 10 animals per concentration). A control group (without anaesthetic) and a complementary group, which was exposed to ethanol, were also evaluated. After exposure to the anaesthetic, the fish were transferred to clean water to assess recovery. The mortality and feeding behaviour of the fish were then observed for 48 h after exposure to the oils. All of the essential oils produced an anaesthetic effect on A. ocellaris. The 27, 70 and 500 μL L^?1 concentrations of clove, mint, and camphor oils promoted surgical anaesthesia after 310.5, 312.0, and 535.0 s (medians) respectively. The recovery times of fish exposed to these same concentrations were 396, 329.5 and 229 s respectively. The decision of which oil to use is dependent on the management situation and the consideration of the induction and recovery times of each essential oil.     

Inhibitory activity of essential oils from medicinal plants against Pseudomonas sp. isolated from aquatic environments

This study focuses on the evaluation of the antimicrobial activity of essential oils (EOs) from medicinal plants against Pseudomonas sp isolated from aquatic environments, and proves its controlling efficacy on Pseudomonas aeruginosa‐Dahp1 using Confocal Laser scanning Microscopy. Twenty‐seven Pseudomonas spp. (Ps ₁–Ps ₂₇) were isolated using King's B medium and the selected P. aeruginosa‐Dahp1 was confirmed using 16S r‐DNA methods. Antimicrobial activity of 10 EOs was determined using agar well and disc diffusion methods. Ten EOs were extracted with six solvents and the crude extracts were tested against the Pseudomonas spp. through disc diffusion method. Among 10 EOs tested, maximum inhibitory activity was noted in EOs of Wringtia tinctoria against the P. aeruginosa‐Dahp1 in the disc diffusion method. The MIC values (concentrations were expressed in Weight/Volume) of the EOs range from 0.5 to 9.054 mg mL^?1. The EO of W. tinctoria showed the maximum activity at the concentration (w/v) of 2.060 mg mL^?1. The EO of W. tinctoria with chloroform extract showed the maximum inhibition against P. aeruginosa‐Dahp1. The CLSM proves the control and viability of 1 × 10⁵ CFU mL^?1 of P. aeruginosa‐Dahp1 at a lower concentration (2.720 mg) of EOs with chloroform extracts of W. tinctoria. This study pivots for designing of new drugs using EOs of W. tinctoria against Pseudomonades in the aquaculture sites.     

Effect of miconazole nitrate on immunological response and its preventive efficacy in Labeo rohita fingerlings against oomycetes Saprolegnia parasitica

This study evaluated the effect of sublethal doses of antifungal drug miconazole nitrate (MCZ) on immunological responses and its role as a prophylactic drug against S. parasitica in Labeo rohita fingerlings. Fish were fed with sublethal doses of MCZ, that is, T1—6.30 mgMCZ kgBW^?1, T2—12.61 mgMCZ kgBW^?1 and T3—25.22 mgMCZ kgBW^?1, and sampling was done at different time intervals for 240 hr. Immunological parameters viz. lysozyme activity, oxygen radical production and plasma antiprotease activity showed significant enhancement (p < 0.05) in fish fed with T2 and T3 doses. Expression of immune‐relevant genes such as TLR‐22 and β2‐M showed significantly higher expression at 6 hr and 24 hr of sampling in both liver and head kidney. However, these genes showed a downregulation after 120 hr of sampling in both the tissues. Preventive efficacy study showed that single dose of MCZ provides protection against oomycetes up to the fourth day of infection. Significantly higher mortality was observed in control diet‐fed fish as compared to fish fed with MCZ medicated diet. Thus, it can be concluded that the MCZ can act as a potent antifungal agent for preventing oomycetes infection as well as to enhance the immune response.     

Effects of Dietary Savory and Myrtle Essential Oils on Growth,Survival, Nutritional Indices,Serum Biochemistry,and Hematology of Farmed Rainbow Trout,Oncorhynchus mykiss,Fry

The effects of diets containing different levels of essential oils from savory Satureja khuzestanica Jamzad and myrtle Myrtus communis L and an immunity complement were investigated on growth, survival, nutritional indices, serum biochemistry, and hematology of farmed rainbow trout, Oncorhynchus mykiss, fry (n = 4500; 5 ± 2 g). The essential oils were dosed as 300 and 500 mg/kg diet. After 60 d, the fish fed 300 mg/kg of the essential oils showed the highest counts of white blood cells (P < 0.05). The same treatment revealed the greatest levels of hematocrit, total protein, and albumin, but glucose and cholesterol values significantly decreased compared with the control (P < 0.05). Feeding fish with 300 mg/kg of the essential oils led to highest records of specific growth rate and survival rate but the lowest feed conversion ratio (FCR), and those fed with immunity complement attained the lowest values of growth parameters. The fry fed with immunity complement and 300 mg/kg of myrtle essential oil achieved the highest and lowest FCR values, respectively. The results indicate that dietary supplementation of 300 mg/kg of the above essential oils exert positive impacts on the growth performance, feeding efficiency, and hematological factors in O. mykiss.     

Fertilization by refrigerator stored sperm of the Indian major carp, Labeo calbasu (Hamilton, 1822)

This study reports on the spermatological properties, and on the development of a protocol for refrigerator storage (4°C) of Labeo calbasu (Hamilton, 1822) sperm for artificial breeding. Volume, motility, concentration and pH of the freshly collected sperm were 2.21 ± 0.53 (μL g^?1 of fish weight) (mean ± SD), 95 ± 1 (%), 1.93 ± 0.44 × 10⁹ (cells mL^?1) and 7.56 ± 0.17 respectively. Sperm activation was evaluated at different osmolalities of NaCl solution, and motility ceased completely when osmolality of the extender was ≥287 mOsmol kg^?1. Sperm retained motility for 24, 72 and 108 h, after refrigerator storage when sperm were undiluted, suspended in Alsever's solution and suspended in Alsever's solution containing 5% methanol respectively. Fertilization rate of fresh eggs with sperm stored at 4°C in Alsever's solution and Alsever's solution containing 5% methanol was 77% and 60% with a hatching rate of 60% and 43% respectively. The fertilization and hatching success of the stored sperm suggests potential to use refrigeration for transporting genetic material to hatcheries for artificial breeding of L. calbasu in Bangladesh.     

Histological alterations in gills of shrimp Litopenaeus vannamei in low‐salinity waters under different stocking densities: Potential relationship with nitrogen compounds

Two experimental modules with different stocking densities (M1 = 70 and M2 = 120 shrimp /m²) were examined weekly over a culture cycle in tanks with low‐salinity water (1.9 g/L) and zero water exchange. Results showed survival rates of 87.7 and 11.9% in M1 and M2, respectively. Water temperature, pH, dissolved oxygen, electrical conductivity and chlorophyll a were not significantly (p > .05) different between modules. In contrast, the concentrations of nitrogen compounds were significantly (p < .05) different between modules, except nitrite‐N (M2 were 2.31 ± 1.38 mg/L N‐TAN, 0.18 ± 0.49 mg/L N‐NO₂^? and 6.83 ± 6.52 mg/L N‐NO₃^?; in M1: 0.97 ± 0.73 mg/L N‐TAN, 0.05 ± 0.21 mg/L N‐NO₂^? and 0.63 ± 0.70 mg/L N‐NO₃^?). When waters of both modules reached higher levels of ammonia and nitrite, histological alterations were observed in gills. The histological alterations index (HAI) was higher in M2 (5‐112) than in M1 (2‐22).     

Protective effects of a synbiotic against experimental Saprolegnia parasitica infection in rainbow trout (Oncorhynchus mykiss)

A combination of probiotics and prebiotics as synbiotics allows assessing their synergistic effects. This study evaluated the effects of a synbiotic supplement on growth performance, haematological parameters and resistance to Saprolegnia parasitica in rainbow trout, Oncorhynchus mykiss (Walbaum) fingerlings. Fish fed a dietary synbiotic in three levels of 0.5, 1.0 and 1.5 g kg^?1 thrice a day. The fingerlings were challenged with Saprolegnia parasitica after 60 days post feeding and their mortalities recorded up to 15 days. The fingerlings at all three experimental treatments showed significant (P < 0.05) increases in final mean weights and specific growth rates (SGR). The best feed conversion ratio (FCR), feed conversion efficiency (FCE) and maximum survival rate were also obtained by the fish fed 1.0 g synbiotic kg^?1 diet. Furthermore, supplementation with synbiotic significantly increased blood factors at all treatments. After challenges with Saprolegnia parasitica, the synbiotic‐fed groups showed significantly higher survival rates compared with the control group. These results reveal that a dietary synbiotic of 1.0 g kg^?1 fed for 60 days leads to increased growth performance and survival rate as well as improved feeding efficiency in rainbow trout fingerling, rendering them more resistant against infection by Saprolegnia parasitica.     

Growth parameters,innate immune response and resistance to Listonella (Vibrio) anguillarum of Dicentrarchus labrax fed carvacrol supplemented diets

The research was aimed to assess the effect of dietary carvacrol (0.025% and 0.05%) on sea bass (Dicentrarchus labrax) growth, immune response and resistance to Listonella anguillarum. Fish (69.2 ± 0.22 g) were fed the experimental diets for 9 weeks. Dietary carvacrol did not negatively affect fish survival, growth performance, feed intake and feed conversion ratio (P > 0.05) nor carcass yield and viscerosomatic, hepatosomatic and mesenteric fat index (P > 0.05). Serum and head kidney leucocytes were collected after 1, 4 and 8 weeks of feeding. Carvacrol significantly reduced serum proteins, immunoglobulins and lysozyme activity (P < 0.01) and moderately increased phagocytosis and pinocytosis of head kidney macrophages. The release of reactive oxygen species by leucocytes was reduced in carvacrol‐fed fish, even if significantly (P < 0.05) only in those fed 0.05% carvacrol for 1 week. Dietary carvacrol did not significantly affect the aspecific immune response, although a potential antioxidant activity might be speculated. Moreover, feeding carvacrol provided an appreciable resistance to a challenge with L. anguillarum, when a bacterial dose lower than the Lethal Dose₅₀ was used. Cumulative mortality in fish fed 0.025% carvacrol was significantly lower than that of untreated controls (75% Relative Per cent Survival).     

Differential pathogenicity of five Streptococcus agalactiae isolates of diverse geographic origin in Nile tilapia (Oreochromis niloticus L.)

Streptococcus agalactiae is an emerging pathogen of fish and has caused significant morbidity and mortality worldwide. The main objective of this study is to assess whether pathogenic differences exist among isolates from different geographic locations. Nile tilapia (Oreochromis niloticus L.) were administered an intraperitoneal injection of suspension containing USA, Brazil, Honduras, Israel, or Kuwait S. agalactiae isolates at concentrations ranging from 10² to 10⁷ cfu mL^?1. The LD₅₀ values 7 days after challenge were as follows: USA (1.0 × 10² cfu mL^?1), Brazil (1.5 × 10³ cfu mL^?1), Honduras (6.8 × 10³ cfu mL^?1), Israel (1.0 × 10⁴ cfu mL^?1) and Kuwait (7.2 × 10⁵ cfu mL^?1). Fish from all groups exhibited lethargy, anorexia, exophthalmia, corneal opacity, erratic swimming, petechiae and mortality. Opercular clearing and ascites were only found after infection with certain geographic isolates. The findings in this study indicate that S. agalactiae isolates of different geographic origin can cause significant mortalities after experimental challenge and can have different pathogenic capacities. Isolates from the Americas (USA, Brazil and Honduras) were more pathogenic to Nile tilapia than isolates from the Middle East/Asia (Israel and Kuwait).     

Efficacy of garlic (Allium sativum) extract applied as a therapeutic immersion treatment for Neobenedenia sp. management in aquaculture

Garlic, Allium sativum L., extract administered as a therapeutic bath was shown to have antiparasitic properties towards Neobenedenia sp. (MacCallum) (Platyhelminthes: Monogenea) infecting farmed barramundi, Lates calcarifer (Bloch). The effect of garlic extract (active component allicin) immersion on Neobenedenia sp. egg development, hatching success, oncomiracidia (larvae) longevity, infection success and juvenile Neobenedenia survival was examined and compared with freshwater and formalin immersion. Garlic extract was found to significantly impede hatching success (5% ± 5%) and oncomiracidia longevity (<2 h) at allicin concentrations of 15.2 μL L^?1, while eggs in the seawater control had >95% hatching success and mean oncomiracidia longevity of 37 ± 3 h. At much lower allicin concentrations (0.76 and 1.52 μL L^?1), garlic extract also significantly reduced Neobenedenia infection success of L. calcarifer to 25% ± 4% and 11% ± 4%, respectively, compared with 55% ± 7% in the seawater control. Juvenile Neobenedenia attached to host fish proved to be highly resistant to allicin with 96% surviving 1‐h immersion in 10 mL L^?1 (15.2 μL L^?1 allicin) of garlic extract. Allicin‐containing garlic extracts show potential for development as a therapy to manage monogenean infections in intensive aquaculture with the greatest impact at the egg and larval stages.     

A3α‐peptidoglycan extracted from Bifidobacterium sp. could enhance immunological ability of Apostichopus japonicus

To assess the effects of A3α‐peptidoglycan (A3α‐PG) extracted from Bifidobacterium sp. on the immune response and disease resistance of sea cucumber, different concentrations (0, 0.5, 5 and 50 mg mL^?1) of A3α‐PG suspensions were used to perform hypodermic injection on Apostichopus japonicus, followed by a Vibrio splendidus challenge. Total coelomocyte count (TCC), phagocytosis activity and activities of four immunological enzymes in both cell‐free coelomic fluid (extra‐cellular, EC) and coelomocyte lysate supernatant (intracellular, IC), including acid phosphatase (ACP), alkaline phosphatase (ALP), superoxide dismutase (SOD) and peroxidase (POD), were measured at 2, 6, 14 and 24 h post injection (hpi). The TCC was not significantly affected (P > 0.05) by A3α‐PG, ranging from 1.84 × 10⁶ to 3.53 × 10⁶ cells mL^?1. The coelomocyte phagocytosis activity was significantly activated (P < 0.05) in all the A3α‐PG treatments, whereas no significant difference was observed between them except 24 hpi (P > 0.05). The EC‐ACP activity in the 5.0 mg mL^?1 treatment increased significantly (P < 0.05) at all sampling times, while the IC‐ACP activity in the 50 mg mL^?1 treatment increased significantly (P < 0.05) at 2 hpi. Also, the 5.0 mg mL^?1 treatment had significant (P < 0.05) increase in the EC‐ALP activity within 14 hpi and the EC‐POD activity at 2 hpi, respectively, while significantly (P < 0.05) enhanced IC‐ALP and IC‐POD activities were observed in the 50 mg mL^?1 treatment within 6 hpi and at 2 hpi, respectively. Only the 5.0 mg mL^?1 treatment showed significant (P < 0.05) increase in the EC‐SOD activity at 2 hpi and IC‐SOD activity within 14 hpi, respectively. The challenge test showed that the animals treated with 50 mg mL^?1 of A3α‐PG had notably lower cumulative mortality after 14 days following V. splendidus exposure. All together, these results suggest that A3α‐PG could positively enhance immune response that effectively promotes the health status of A. japonicus against V. splendidus infection.