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This study investigated the synergistic effects of oxolinic acid (OA) combined with oxytetracycline (OTC) on white shrimp (Penaeus vannamei). Disk diffusion susceptibility testing was performed to analyze the sensitivity of Vibrio alginolyticus and Vibrio parahaemolyticus to different concentrations of OA and OTC. The results revealed that 50 mg OA/L combined with 50 mg OTC/L exhibited stronger antibacterial effects on V. alginolyticus and V. parahaemolyticus. The results of in vitro tests indicated that cotreatment with OA and OTC significantly reduced superoxide anion production and phenoloxidase activity, but not phagocytic activity. Subsequently, feeding trials were performed to investigate the immunomodulatory effects and bioaccumulation of dietary OA combined with OTC on shrimp. The healthy shrimp (15.13 ± 1.02 g) were divided into four groups: control, 100 mg OA/kg combined with 50 mg OTC/kg, 50 mg OA/kg combined with 100 mg OTC/kg, and 50 mg OA/kg combined with 50 mg OTC/kg. The shrimp were sampled to determine innate immunity parameters and residual OA and OTC levels in the muscle during a 28-day feeding regimen; the shrimp were fed the experimental diet from day 1 to day 5 and a commercial diet from day 6 to day 28. Residual OA levels were considerably higher in the group fed 100 mg OA/kg combined with 50 mg OTC/kg compared with the other groups and peaked on day 4. The residual OA levels of all the groups were below the detection limit after without providing OA. The residual OTC levels of the group fed 50 mg OA/kg combined with 100 mg OTC/kg were considerably higher from day 1 to day 4. The residual OTC levels in all the groups decreased rapidly and could not be detected on day 28. The administration of 50 mg OA/kg combined with 100 mg OTC/kg exerted the least effect on the white shrimp. Moreover, the survival rates of the treatment groups after the V. parahaemolyticus challenge were higher than those of the control group, especially the group fed 50 mg OA/kg combined with 100 mg OTC/kg. This result indicated that the synergistic effects of dietary OA and OTC are safe and effective. Combination therapy is a new method of antibiotic use in aquaculture.

This study investigated the effect of the moringa (Moringa oleifera) leaf extract and Lactobacillus acidophilus individually or combined on growth performance, enzyme activity, intestinal and hepatopancreatic histology, intestinal microbiota, immune response, and resistance against Vibrio alginolyticus and Vibrio parahaemolyticus in whiteleg shrimp (Penaeus vannamei). Six diets were formulated: three diets without L. acidophilus containining 0 (control, ME0), 2.5 (ME2.5), and 5.0 g/kg of moringa (ME5.0) and the same three diets containing L. acidophilus at 1 × 10

Nile tilapia (Oreochromis niloticus) is an important food fish species that is mainly cultivated in tropical and subtropical countries. However, microbial diseases have created various difficulties for this industry. The fundamental prerequisite for tackling disease outbreak prevention and disease resistance is to know how hosts' immune responses against invading microbes are initiated. Toll-like receptors (TLRs) are vital pattern recognition receptors and play pivotal roles in the cellular innate immunity defense that is able to recognize pathogen-associated molecular patterns (PAMPs). In this study, Oreochromis niloticus TLR23 (OnTLR23) was cloned and bioinformatic analyses revealed that OnTLR23 is not an ortholog of mammalian TLR13 as previously suggested. The basal transcript level of OnTLR23 was found to be higher in the immune-related organs and was upregulated in the spleen and/or head kidney following Aeromonas hydrophila, Streptococcus agalactiae or poly I:C injections, and increased in the melanomacrophage-like tilapia head kidney (THK) cell line after LPS and zymosan stimulation. Furthermore, we demonstrated for the first time that OnTLR23 locates mainly in the intracellular region in fish cells and the constitutively active form of OnTLR23 promotes the expression of molecules related to antigen presentation, proinflammatory cytokines, antimicrobial peptides and type I interferon in THK cells. A co-immunoprecipitation assay revealed that OnTLR23 can interact with both OnMyD88 and OnTRIF, but not with OnTIRAP. A luciferase assay showed that the NF-κB activity was not elevated in the OnTLR23 overexpressed THK cells after treatment with ligand for TLR13 as well as other known purified bacterial-derived ligands of TLRs. Taken together, OnTLR23 is likely to recruit OnMyD88 and OnTRIF as adaptors to induce the expression of various effectors in melanomacrophages, but its corresponding ligand is an issue awaiting further investigation.

Toll-like receptors (TLRs) are evolutionarily conserved proteins of pattern recognition receptors (PRRs) and play a crucial role in innate immune systems recognition of conserved pathogen-related molecular samples (PAMPs). We identified and characterized TLR18 from Nile tilapia (Oreochromis niloticus), OnTLR18, to elucidate its role in tissue expression patterns, modulation of gene expression after microbial challenge and TLR ligands, subcellular localization in fish and human cells, and the possible effectors TLR18 induces in a melanomacrophage-like cell line (tilapia head kidney (THK) cells). OnTLR18 expression was detected in all tissues examined, with the highest levels in the intestine and the lowest in the liver. OnTLR18 transcript was up-regulated in immune-related organs after bacterial and polyinosinic-polycytidylic acid (poly I:C) challenges and in the THK cells after lipopolysaccharide (LPS) stimulation. In transfected THK and human embryonic kidney (HEK) 293 cells, OnTLR18 localizes in the intracellular compartment. OnMyD88 and OnTRIF, but not OnTIRAP, were co-immunoprecipitated with OnTLR18, suggesting that the former two molecules are recruited by OnTLR18 as adaptors. The constitutively active form of OnTLR18 induced the production of pro-inflammatory cytokines, type I interferon (IFN), and antimicrobial peptides such as tumor necrosis factor α, interferon (IFN) d2.13, tilapia piscidin (TP)2, TP3, TP4, and hepcidin in THK cells. Our results suggest that OnTLR18 plays an important role in innate immunity through initiating nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and IFN signaling pathways via OnMyD88 and OnTRIF and induces the production of various effectors in melanomacrophages.

Anthropogenic climate change is known to be an increased stress that affects aquatic animal behavior and physiological alternations, which can induce the animal's death. In order to known whether the extracted acetyl-xylogalactan function on the regulation of the external high temperature induced death, we first selected the mammalian cell line "RAW 264.7" used in the previous experiment to evaluate the extracted acetyl-xylogalactan function. We aimed to evaluate the effects of the acetyl-xylogalactan on the RAW 264.7 macrophages and Nile Tilapia stress factor expression under the heat environment. In the in vitro cell observation, we assessed the cell survival, phagocytic activity, intracellular Ca

The water environment plays an important role in animal physiology. In this study, we sought to evaluate the effect of the acid environment on the

This study investigated the effects of two probiotics, namely Lactobacillus paracasei and Bifidobacterium longum, as feed additives on growth performance, nonspecific immunity, immune-related gene expression, and disease resistance against Vibrio parahaemolyticus in Penaeus vannamei. The experimental diets were prepared using L. paracasei and B. longum at concentrations of 10

This study investigates the effects of Phyllanthus amarus extract (PAE) on immune responses, growth, and resistance to Vibrio alginolyticus in white shrimp (Litopenaeus vannamei). In vitro PAE treatment did not alter the cell viability of haemocytes and significantly enhanced immune parameters such as phenoloxidase (PO) activity, phagocytic activity, and superoxide anion (O2−) production. We conducted two feeding trials to examine the effects of PAE on the growth, disease resistance, and innate immune parameters of white shrimp. In the first in vivo trial, shrimps (4.01 ± 0.03 g) were fed a diet containing 0 g (control), 10 g (PAE10), 20 g (PAE20), or 40 g (PAE40) of PAE per kilogram of feed for 56 days. After the feeding period, the PAE20 group showed a significantly higher weight gain and specific growth rate than shrimp fed the control diet. Furthermore, after challenge with V. alginolyticus, shrimp fed a diet containing PAE showed significantly higher survival than those fed the control diet. The second in vivo trial (28 days) was performed to identify the mechanisms of enhanced immunity in PAE-fed shrimp. Shrimp fed the PAE20 diet generally had the highest total haemocyte count, PO activity, phagocytic activity, and O2− production, followed by the PAE40 and PAE10 groups. Thus, our results suggest that administration of 20 g of PAE per kilogram of feed can enhance immunity, growth, and resistance to V. alginolyticus in white shrimp.

In this study, water extracts of the red seaweed Sarcodia suiae were obtained using solid-liquid extraction (SLE) or pressurized liquid extraction (PLE) methods. The extracts were used to investigate immunostimulatory activity by measuring the phagocytic activity of Nile tilapia (Oreochromis niloticus) hepatic and splenic macrophages and the tilapia head kidney (THK) cell line, and modulation of immune-related genes in primary head kidney (HK) cells and THK cells. At 10 μg/ml, both extracts promoted the proliferation of hepatic and splenic macrophages. Expression levels of proinflammatory cytokines (IL-1β and IL-8), antimicrobial peptides (TP2 and TP4), and pattern recognition receptors (TLR5) were elevated in SLE extracts-treated primary HK leukocytes. Similarly, IL-1β, IL-8, and TNFα expression was also induced by SLE extract in THK cells. Phagocytic activity in primary HK cells and THK cells was induced by SLE extract 12 h and 24 h post-stimulation, while PLE extract only induced phagocytic activity in THK cells at early time points. SLE extract (100 μg/g body weight) increased the expression of IL-1β, IL-8, TNFα, TP2, TP4, TLR2 and TLR5 in the spleen and immunoprotective efficiency against Streptococcus agalactiae infection. Taken together, these results show that S. suiae can differentially stimulate the immune response of tilapia in vitro and in vivo and could potentially be used as an immunomodulator in tilapia culture.

The white shrimp Litopenaeus vannamei is one of the most commercially important aquatic species. However, intensive farming to meet economic demands has been linked to animal stress and subsequent disease outbreaks. In this study, we explore the immunomodulatory effects of Sargassum horneri hot-water extract (SHE) on the immune parameters of L. vannamei. Hemocytes incubated in vitro with 10 mg/ml of SHE showed the highest response in phenoloxidase (PO) activity and reactive oxygen species production. In the in vivo trials, shrimp were fed diets containing 0 (control), 2.5 (SHE 2.5), 5.0 g (SHE 5.0), and 10 g (SHE10.0) of lyophilized SHE per kg feed for four weeks, after which immune parameters were measured. Group SHE10.0 had the highest total hemocyte count, reactive oxygen species production, and phagocytic rate during the 4-week feeding period, followed by groups SHE5.0 and SHE2.5. However, group SHE5.0 showed a better growth performance among all examined groups after four weeks of rearing. Furthermore, gene expression analysis revealed that L. vannamei fed diets containing 0.5% SHE over 28 days showed significant modulation of 11 immune-related genes, including prophenoloxidase I, prophenoloxidase II, peroxinectin, α2macroglobulin, clotting protein, lysozyme, superoxide dismutase, glutathione peroxidase, penaiedin2-4, and crustin. Combined, these results show that SHE is a functional feed additive that can be applied orally to enhance innate immunity and growth performance of white shrimps.

This study discussed the effects of two types of lactic acid bacteria, Lactobacillus reuteri (L. reuteri) and Pediococcus acidilactici (P. acidilactici), on the growth and nonspecific immunity of Penaeus vannamei (P. vannamei) and developed probiotic applications for shrimp cultivation. This study incorporated two types of lactic acid bacteria in shrimp feed through spraying. The shrimps were grouped according to the type and concentration of effective bacteria incorporated into their feed. This research was separated into 3 individual feeding treatment as control, L. reuteri (Lr groups) and P. acidilactici (Pa groups). The shrimp was feeding with 10

Mislabelling the geographic origin of same-species aquaculture products is difficult to identify. This study applied untargeted small-molecule fingerprinting to discriminating between Atlantic salmon originating from Chile and Norway. The acquired liquid chromatography-high-resolution mass spectrometry data from Chilean (n = 32) and Norwegian (n = 29) salmon were chemometrically processed. The partial least squares discriminant analysis (PLS-DA) models successfully discriminated between Chilean and Norwegian salmon at both positive and negative ionisation modes (R

Trivalent arsenic (As (III)) contamination in the marine environment can produce adverse effects in crustaceans. The present study investigated the chronic toxicity of As (III) in white shrimp (Penaeus vannamei) by analyzing the tissue bioaccumulation and non-specific immune responses. Shrimps were exposed to 0 (control), 50, 500, and 2500 μg/L of As (III) for 21 days. The results showed that the hepatopancreas was the main tissue of arsenic accumulation in white shrimp. The cumulative concentration of total arsenic and inorganic arsenic but not arsenobetaine was positively correlated with the exposure concentration. In vitro As (III) treatment (0-2500 μg/L) with haemocytes isolated from healthy shrimp did not cause the cytotoxicity, but this arsenic treatments inhibited the phagocytic rate and O

This study investigated the effects of guava leaf extract (GLE) on immune responses, growth performance, and resistance to Vibrio parahaemolyticus in white shrimp (Penaeus vannamei). To examine the effect of GLE on the immune response of white shrimps, they were treated with various concentrations of GLE on hemocyte (in vitro) and were orally administered (in vivo) feed containing various concentrations of 0, 1, 5, and 10 g kg−1 GLE (control, GLE1, GLE5, and GLE10, respectively) for 28 days. Furthermore, their growth performance was evaluated for 56 days. In a separate experiment, the shrimps were challenged with V. parahaemolyticus injection after 7 days of culture. In vitro experiments indicated that GLE is nontoxic and can activate immune response. In vivo experiments revealed that the GLE5 led to the highest total hemocyte count, phenoloxidase activity, phagocytic activity, and superoxide anion production and the highest upregulation of lipopolysaccharide, β-1,3-glucan-binding protein, peroxinectin, lysozyme, crustin, penaeidin 2, penaeidin 3, clotting protein, superoxide dismutase, and glutathione peroxidase. Moreover, better growth performance was observed in the GLE groups, with GLE5 exhibiting the highest specific growth rate, weight gain, and feed conversion rate. In addition, GLE5 enhanced resistance to V. parahaemolyticus, with a survival rate of 72.27%. In conclusion, GLE was found to be effective in enhancing nonspecific immune response and growth performance and in reducing V. parahaemolyticus infection in white shrimp.

In this study, the effects of acetyl-xylogalactan extracted from Sarcodia suieae on RAW 264.7 macrophage polarisation were evaluated. This extracted acetyl-xylogalactan had a monosaccharide composition of 91% galactose and 9% xylose, with polysaccharide and acetyl contents of 80.6% and 19.3%, respectively. MALDI–TOF mass spectrometry and NMR spectroscopy revealed the molecular weight of the acetyl-xylogalactan to be 88.5 kDa. After acetyl-xylogalactan treatment, RAW 264.7 macrophage polarisation was noted, along with enhanced phagocytic ability. Furthermore, the Cell Counting Kit-8 (CCK-8) assay was performed and the results demonstrated non-significant alteration in lactate dehydrogenase levels in the treated cells. Next, interleukin (IL) 1β, TNF, and Malt-1 expression in RAW 264.7 macrophages treated with the S. suieae acetyl-xylogalactan was investigated through real-time quantitative polymerase chain reaction, and the results demonstrated that S. suieae acetyl-xylogalactan induced IL-1β and Malt-1 expression. RNA sequencing analysis results indicated the S. suieae acetyl-xylogalactan positively regulated cytokine production and secretion, protein secretion, and response to IL-1 activation, based on the observed GO terms. The predicted target genes in the GO enrichment analysis were found to upregulate NF-κB signalling and M0 to M1 macrophage conversion through the observed cytokine production. Thus, acetyl-xylogalactan can positively regulate RAW 264.7 macrophage polarisation.

This study investigates the effects of three medicinal herbal extracts, namely Bidens pilosa (BPE), Lonicera japonica (LJE), and Cyathula officinalis (COE), on nonspecific immune parameters of cobia (Rachycentron canadum) in vitro and in vivo. During in vitro tests, BPE treatment increased reactive oxygen species (ROS) production in a dose-dependent manner in primary head kidney leukocytes. Similarly, ROS production rates were enhanced by LJE (50 and 100 mg/ml) and COE (100 mg/ml). This suggests that these three herbal extracts possess immunostimulating properties. We then conducted two feeding trials to examine the effects of these three herbal extracts on growth and innate immune parameters of cobia, and sought an optimal dietary supplementation proportion required for activating the non-specific immune responses. In the first trial, we supplemented the diet with 1, 5, or 10% of the individual extracts. After a ten-week feeding trial, no negative impacts on weight gain, feed conversion rate, and survival rate were observed in fish offered experimental diets. Further, ROS production, phagocytic capacity of the head kidney leukocytes, and serum lysozyme activity were enhanced by differing degrees in fish fed the herbal extracts compared to fish in the control group. A similar albumin/globulin ratio was seen between each experimental group and the control group regardless of the type and dose of herbal extract used, indicating these medicinal herbal extracts are safe for cobia. We then performed a 30-day feeding trial with lower extract concentrations (1, 3, and 5% of the diet) to identify dose responses in cobia at various time points so that we could establish a cost-effective manner of administering the three extracts for cobia. All BPE fed fish had higher ROS production compared to the control group, while phagocytosis rate and index were simultaneously raised in only the BPE30 group (3% BPE). Immune parameters such as ROS production, phagocytic rate, and serum lysozyme activity were triggered when fish received 30 g LJE per kg of feed. However, ROS production only increased in the LJE10 group (1% LJE) on day 30 and was not enhanced in the LJE50 group (5% LJE). Additionally, although the phagocytic rate and phagocytic index were induced in the LJE50 group, serum lysozyme activity was not elevated in this group (LJE50) at any time point examined. ROS production was greatly improved in all COE fed groups, but only the COE30 group (3% COE) showed prolonged enhanced phagocytic rate over the 30-day feeding trial.

We investigated the antimicrobial properties and the effects of Rheum officinale extract (ROE) on nonspecific immune parameters of orange-spotted grouper (Epinephelus coioides) in vitro and in vivo. The in vitro analysis was conducted by treating grouper primary head kidney leukocytes with various concentrations of ROE. The phagocytic rate of the leukocytes was elevated in a dose-dependent manner from 0.01 to 0.1 mg/ml, but decreased with higher concentrations of ROE (0.5 and 1.0 mg/ml). The production of reactive oxygen species (ROS) was strongly enhanced in a dose-dependent manner by treatment with ROE doses of 0.1–10.0 mg/ml. However, morphological changes (e.g., rounding and shrinkage of cells, chromatin condensation, fragmentation, and appearance of apoptotic bodies) were observed in the leukocytes after incubation with higher concentrations of ROE (1.0 and 10.0 mg/ml). A 28-day feeding trial was performed to assess the impact of dietary administration of ROE on grouper innate immunity parameters. Fish were fed with feed supplemented with 0, 0.1, 1.0, or 5.0 g ROE per kg of feed. The phagocytic activity of the animals' leukocytes was significantly elevated in all ROE-fed groups on day 1 and in groups fed with ROE at 0.1 or 1.0 g/kg on day 14. Production of ROS was substantially increased on day 1 in fish fed with ROE at 1.0 and 5.0 g/kg, but decreased steadily later on. The ability to generate ROS increased steadily until day 7 in fish fed the lowest concentration of ROE (0.1 mg/ml), but decreased thereafter. ROE showed excellent antibacterial activity against six pathogens of aquatic animals: Vibrio parahaemolyticus, V. vulnificus, V. alginolyticus, V. carchariae, Aeromonas hydrophila, and Edwardsiella tarda. The minimum inhibitory and bactericidal concentrations of measured ROE-derived anthraquinones were 10.57–84.53 μg/ml and 10.57–169.05 μg/ml, respectively.

This study was conducted to examine the combinatory effects of β-glucan and oxytetracycline (OTC) on hybrid giant tiger groupers (Epinephelus fuscoguttatus × Epinephelus lanceolatus). In vitro tests, OTC significantly reduced superoxide anion production and phagocytic activity in primary head kidney leukocytes. However, this suppressive effect was alleviated by co-treatment with β-glucan. Subsequently, feeding trials were performed to investigate the potential immunomodulatory effects of dietary β-glucan alone or in combination with OTC on groupers. A total of 210 healthy groupers (368.00 ± 51.03 g) were divided into six groups. Group 1 was the control group, group 2 (BG) received 5 g β-glucan per kg feed weight, groups 3–5 received 5 g/kg β-glucan in combination with 10, 30, or 50 mg OTC/kg fish weight/day (groups M1, M2, and M3, respectively), and group 6 (O) received 50 mg OTC/kg fish weight/day. Fish were sampled to determine the innate immunity parameters and residual OTC levels in the muscle tissue during a 28-day feeding regimen. Residual OTC levels were considerably higher in groups M3 and O compared with the other groups, and peaked on day 14. This was followed by a slight decrease on day 28, despite a continuous supply of OTC. Notably, fish fed with OTC alone had significantly decreased phagocytic rates and superoxide anion production observed in head kidney leukocytes, as well as poorer protection against Vibrio alginolyticus infection. These immunosuppressive effects were not observed in the fish fed with β-glucan in combination with a lower dose of OTC (group M2). Thus, these data suggest that the combination of dietary β-glucan and OTC exerts synergistic immunostimulating effects that protect groupers from bacterial infection.

This paper examines the health risks of exposure to methylmercury (MeHg) through the consumption of mercury-contaminated seafood in Taiwan, based on the total diet study (TDS) method. Samples of seafood (n = 140) were purchased at fishing harbors or supermarkets and classified into seven categories (pelagic fish, inshore fish, farmed fish, shellfish, cephalopods, crustaceans, and algae). For each sample, we analyzed raw and cooked versions and compared the concentration difference. Total mercury (THg) was detected at the highest rate and in the highest concentrations in pelagic fish, followed by inshore fish and other farmed fish. The average concentration of THg was higher after cooking. In a 75th percentile scenario, the hazard indices for children aged 1 to 3 years and children aged 4 to 6 years were higher than 100% of the provisional tolerable weekly intake. Taking into consideration the risk assessment results, MeHg concentrations, and the nutritional composition of fish, we have provided weekly consumption advisories for children aged 1 to 3 years, children aged 4 to 6 years, and childbearing women aged 19 to 49 years. The weekly consumption advisories for childbearing women are 35 g/week of pelagic fish and 245 g/week of inshore fish based on the risk results from MeHg and the potential benefits from eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) intake.

In this study, we investigated organ accumulation and nonspecific immune response in white shrimp ( Litopenaeus vannamei ) that were exposed to various concentrations of lead (Pb) solution. The concentrations of Pb in the hepatopancreas, haemolymph, and muscle were measured moreover the total heamocyte count, phenoloxidase activity, O 2 − and physiological factors such as glutamic oxaloacetate transaminase (GOT), glutamic pyruvic transaminase (GPT), and haemolymph glucose were detected. The results showed that the hepatopancreas was the main organ of accumulation of Pb in white shrimp and the cumulative concentration of each organ was positively correlated with the experimental Pb concentration and immersion time. By observing GOT and GPT, Pb was found to inhibit phenoloxidase and O 2 − activity and to induce organ injury. Thus, the heavy metal Pb accumulates in the hepatopancreas and haemolymph and affects the crustacean metabolic organ injury (rising of GOT and GPT) further to inhibit nonspecific immune responses.

Toll/interleukin 1 receptor domain-containing adaptor protein (TIRAP) and toll/interleukin 1 receptor-domain-containing adapter-inducing interferon-β (TRIF) are crucial adaptors of signal transduction for the signaling pathways of toll-like receptors (TLRs). TIRAP and TRIF perform an essential function in an antimicrobial immune response; however, their function in Nile tilapia remains unknown. Herein, TIRAP and TRIF from Nile tilapia were identified and functionally characterized. Phylogenetic analysis showed that OnTIRAP and OnTRIF clustered with corresponding homologs from other fish species, with comparable gene structures to those of select vertebrate TIRAP and TRIF genes, respectively. The expression profiles of OnTIRAP and OnTRIF were broadly distributed in the ten tissues investigated, with high transcript levels noticed in immune organs. The transcription levels of OnTIRAP and OnTRIF were upregulated in response to bacterial and poly (I:C) challenges. GFP signals were only detected in the cytoplasmic region of fish cells transfected with OnTIRAP-GFP and OnTRIF-GFP expression plasmids. Moreover, overexpression of OnTIRAP and OnTRIF activated interferon-β (IFN-β) and activator protein 1 (AP1) reporters in HEK 293 cells. Activation of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) reporter was only observed in OnTRIF-overexpressing HEK 293 cells. Furthermore, the results of the co-immunoprecipitation analysis showed that OnTRIF, but not OnTIRAP, was recruited as an adaptor protein by OnTLR25. This study provides the first evidence on the functions of OnTIRAP and OnTRIF in the immune system of Nile tilapia against pathogens and may serve as the basis for further investigations on TLR signaling in fish.

This study investigated the effects of orally administered Plantago asiatica , Houttuynia cordata , and Mentha haplocalyx on the growth and nonspecific immune responses of cobia ( Rachycentron canadum ). The nonspecific immune parameters assessed were weight gain, feed conversion ratio, superoxide anion (O 2 − ) production, superoxide dismutase (SOD) activity, phagocytic rate, phagocytic index, lysozyme activity, serum albumin and globulin, and albumin:globulin (A/G) ratio. The growth experiment indicated that 6-week dietary treatments did not significantly affect on the growth of cobia. Nonspecific immune responses showed that O 2 − production, SOD and lysozyme activity, and phagocytosis were significantly increased after the oral administration of P. asiatica and H. cordata , and the serum albumin:globulin ratio (A/G) gradually decreased. In this study, treatment of the Mentha haplocalyx on the cobia didn't present with the inducing of the phagocytosis ability compared with the treatment of P. asiatica and H. cordata. We suggest that oral administration of the 10 g/kg or 20 g/kg of the P. asiatica and H. cordata is exactly inducing the phagocytosis, ROS production, lysozyme activity and SOD production in the cobia.

This study mainly evaluated the effects of orally administered beta 1,3/1,6-glucan and vitamin C on the nonspecific immune responses of white shrimp ( Litopenaeus vannamei ). In this study, we found that the white shrimp oral administration with 1 g/kg of beta 1,3/1,6-glucan effectively enhanced O 2 − production and phenoloxidase and superoxide dismutase activity. Shrimp were oral administration with 0.2 g/kg of vitamin C presented beneficial nonspecific immune responses and enzyme activity and also observed in the beta 1,3/1,6-glucan treatment groups. Consequently, we compared the alterations in the immune activity between the beta 1,3/1,6-glucan and vitamin C groups and the evidence illustrated that combination of beta 1,3/1,6-glucan and vitamin C presented an additive effect on inducing the nonspecific immune responses of white shrimp.

This study investigated the effect of two steroid hormones on phenoloxidase activity, O2(-) production in the haemocytes, total haemocyte count (THC), superoxide dismutase (SOD) activity, glutamate oxaloacetate transaminase (GOT) activity, glutamate pyruvate transaminase (GPT) activity, and plasma cholesterol concentrations in white shrimps (Litopenaeus vannamei). Phenoloxidase activity, THC and plasma cholesterol concentration in shrimps treated with cortisone and 20-hydroxyecdysone were found to be lower when compared with the control groups. In the observation of O2(-) production, treatment of cortisone and hydroxyecdysone were reducing the activity in the 1st day, but to be undiversified with the elapsed time. By contrast, SOD activity in the hepatopancreas, plasma GOT activity, and GPT activity were significantly increased when compared with the control groups.

This research aims to investigate the non-specific immune response of Taiwan abalone (Haliotis diversicolor supertexta) which was treated with the beta-1,3-1,6-glucan to be observed in the survival impact after the Vibrio alginolyticus infection. The non-specific immune and physiological response of superoxide anion radical (O2(-)), phenoloxidase (PO), phagocytic index (PI), phagocytic rate (PR) and lucigenin-chemiluminescence for reactive oxygen intermediates (ROIs) were enhanced via in-vitro experiment. In the in-vivo experiment, the observed data presented that the haemolymph lysate supernatant (HLS), superoxide dismutase (SOD), glutamate oxalacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) were not significant enhanced, but the total haemocyte count (THC), O2(-), PO, phagocytic index (PI), phagocytic ratio (PR) and other parameters of immune were significantly promoted after treated with beta-1,3-1,6-glucan. In the challenge experiment, the survival rates of abalone in the 40 and 80 μl/ml groups of beta-1,3-1,6-glucan were observed from 6.67% up to 33.33% and 36.67% after injection with Vibrio alginolyticus, respectively.

Marbled eels, Anguilla marmorata (Quoy & Gaimard), cultured in Taiwan exhibited haemorrhage and mortality in January 2012. The severely diseased eels bled from the gills and showed congestion of the central venous sinus of the gill filaments and haemorrhage throughout the body similar to viral endothelial cell necrosis of eel. In this study, a novel polyomavirus (AmPyV) was isolated from the diseased eels using the AMPF cell line established from the pectoral fin of healthy marbled eels. AmPyV was found to encode a long T-antigen orthologous gene. Phylogenetic analysis showed that AmPyV was closely related to Japanese eel endothelial cell-infecting virus. PCR assays revealed AmPyV infection throughout the systemic organs. AmPyV proliferated in the AMPF, EK-1 and EO-2 cells at temperatures 25-30 °C, and the progeny virus yields were 10(7.0) , 10(7.4) and 10(7.7) TCID50 mL(-1) , respectively. The purified virions were icosahedral particles, 70-80 nm in diameter. No clinical signs or mortality was observed among the eels injected with the virus; however, the virus was reisolated from the brain, eyes, kidneys, fins and gills of infected eels 2 month after injection. Our results suggest that AmPyV exhibits a latent infection. Pathogen of the disease needs to study further.

Background Binding of beta 1,3/1,6 glucan of Ganoderma lucidum (G. lucidum) with the receptor results in a series of signal transfers (signalling cascades), which activates the transcription factors for regulating inflammation. Excess cholesterol intake leads to an increase in the distance between fat cells and capillaries, which may cause hypoxia in the fat tissue of obese mice. This hypoxia induces the death of fat cells, resulting in the inflammation of adipose tissue or an increase in the inflammatory gene expression associated with obesity. Methods The current study examined the immunomodulation effect of G. lucidum beta 1,3/1,6 glucan according to immunoglobulin, poly-Ig receptor expression, Nature Killer cell (NK cell) activity, lymphocytes proliferation and cytokines expression. Results Our present study shows that feeding G. lucidum beta 1,3/1,6 glucan to mice induces IgA or IgG expression in the serum and small intestine washing fluid and enhances poly-Ig receptor expression in the small intestine moreover, the observation of the IL-2 and Nature killer cell activity were exchanged. Conclusions The effect of a high-cholesterol diet in the inflammatory response was observed in heart, liver, kidney, spleen, and colon tissues through histopathological evaluations. The presented evidence demonstrates that the inflammation response in the high-cholesterol diet group was much higher than in the other groups and the beta 1,3/1,6 glucan reduces inflammation in obese mice fed a high-cholesterol diet.

Rapid, sensitive, and automatic detection platforms are among the major approaches of controlling viral diseases in aquaculture. An efficient detection platform permits the monitoring of pathogen spread and helps to enhance the economic benefits of commercial aquaculture. Nervous necrosis virus (NNV), the cause of viral encephalopathy and retinopathy, is among the most devastating aquaculture viruses that infect marine fish species worldwide. In the present study, a highly sensitive magnetoreduction assay was developed for detecting target biomolecules with a primary focus on NNV antigens. A standard curve of the different NNV concentrations that were isolated from infected Malabar grouper ( Epinephelus malabaricus) was established before experiments were conducted. The test solution was prepared by homogeneous dispersion of magnetic nanoparticles coated with rabbit anti-NNV antibody. The magnetic nanoparticles in the solution were oscillated by magnetic interaction with multiple externally applied, alternating current magnetic fields. The assay’s limit of detection was approximately 2 × 101 TCID50/ml for NNV. Moreover, the immunomagnetic reduction readings for other aquatic viruses (i.e., 1 × 107 TCID50/ml for Infectious pancreatic necrosis virus and 1 × 106.5 TCID50/ml for grouper iridovirus) were below the background noise in the NNV solution, demonstrating the specificity of the new detection platform.

The lysozyme activity, alternative complement activity (ACH50), respiratory burst, SOD (superoxide dismutase) activity and phagocytic activity of orange-spotted grouper Epinephelus coicoides were examined when the fish were injected intraperitoneally with sodium alginate at 10, 20, 30mgkg-1 and iota-carrageenan at 10, 20, 30mgkg-1, respectively after 24, 72 and 120 h. Serum ACH50 increased directly with dose after 24 and 72 h for both sodium alginate and iota-carrageenan treatments. The fish that received sodium alginate at 20mgkg-1 after 24 and 72 h, and the fish that received iota-carrageenan after 72 and 120 h showed significantly increased respiratory burst, SOD activity and phagocytic activity, respectively. In another experiment, E. coicoides which had been injected individually with sodium alginate and iota-carrageenan at 10, 20, 30mgkg-1, were challenged with Vibrio alginolyticus at 1.8x10(9) colony-forming units (cfu)fish-1 and then placed in seawater of 33 per thousand. The survival of fish that received sodium alginate at 20mgkg-1, and the fish that received iota-carrageenan at 30mgkg-1 was significantly higher than that of fish which received saline and the control fish after 48 h as well as at the termination of the experiment (120 h after the challenge). It is therefore concluded that E. coicoides which received sodium alginate at 20mgkg-1 or iota-carrageenan at 30mgkg-1 increased the non-specific immune response and resistance from V. alginolyticus infection.

Wound healing is a complex process involving soluble mediators, blood cells, extracellular matrix, and parenchymal cells in a response that occurs after surgical procedure or traumatic injury. The present study aims to investigate the ROS producing from the injury that involved in the wound healing using the ZFL (zebrafish liver cell) and tilapia partial hepatectomy model. In the ZFL, we observed that while over-inhibition of NADPH activity leading to reduce the wound healing moreover, experiment of the oxidative stress by the extracellular hydrogen peroxide exactly presented to increase the PCNA, BrdU and Ki-67 histopathological repair response of tilapia liver follow partial hepatectomy. We conclude that over inhibition of the NADPH oxidase by DPI may reduce the cell even the tissue in the progress of healing after the injury.

Reactive oxygen species (ROS) may act as signaling molecules in the physiology responses and the present study aims to investigate the effect of extracellular hydrogen peroxide on macrophages cellular response. The results obtained in the present study showed that the extracellular hydrogen peroxide affectively alter the membrane potential of the cell membrane and ion exchange channels in the cell membrane through intracellular NAD turnover that may lead to an intracellular calcium ion concentration alteration and subsequently induce the downstream signal activation.

This study was conducted to investigate the effect of katuk (Sauropus androgynus L. Merr) on growth, non-specific immune and diseases resistance against Vibrio alginolyticus in grouper (Epinephelus coioides). Grouper juveniles (mean weight 10.97 ± 1.99 g, and length 9.67 ± 0.33 cm) were separated into four groups and cultivated in 100-L tank. Each group was fed with diets containing 0, 1.0, 2.5 and 5.0 g/kg herbs diet twice daily. Fish were sampled for non-specific immune parameters at 0, 1, 2, 4, 7, 14 and 30 days. Results showed that fish received S. androgynus at 1.0 and 2.5 g/kg diets affected the growth and non-specific immune responses. Weight gain, specific growth rate, respiratory burst activity, phagocytosis and reactive oxygen species significantly increased in fish administered with 1.0 and 2.5 g/kg S. androgynus diets. The mortality rate after V. alginolyticus challenge decreased in fish fed with 1.0 g/kg S. androgynus extract. Thus, this study indicated that administration of grouper with S. androgynus supplemented diets can affect the growth performances, diseases resistance and enhances non-specific immune responses.

Ependymal radial glial cells, also called tanycytes, are the predominant glial fibrillary acidic protein (GFAP)- and vimentin (VIM)-expressing cells in fish ependyma. Radial glial cells have been proposed to be neural stem cells but their molecular expression is not well understood. Previous studies revealed that fish neural progenitor and neural stem cells have A2B5, a marker for oligodendrocyte progenitor cells (OPCs). In this study, an A2B5(+) cell line, SPB, was isolated from the brain of the teleost Trachinotus blochii and characterized. SPB cells usually grew as polygonal epithelial cells, but at high density, long processes were commonly observed. Using immunocytochemistry, SPB cells were shown to exhibit oligodendrocyte markers such as galactocerebroside and Olig2, and radial glial cell markers such as brain lipid-binding protein, GFAP, Sox2, and VIM. SPB cells were also observed to have DARPP-32, a marker for tanycytes in mammals, and primary cilia. RT-PCR additionally revealed expression of bone morphogenetic protein 4, connexin35, Noggin2, and proteolipid protein in SPB cells. Results of this study suggest that SPB cells are OPCs that can display tanycyte characteristics. Fish tanycytes can be neural stem cells suggesting that SPB cells are neural stem cells. SPB is the first fish cell line showing primary cilia and markers for both OPCs and tanycytes.