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  1. Lee PT, Yamamoto FY, Low CF, Loh JY, Chong CM
    Front Immunol, 2021;12:773193.
    PMID: 34975860 DOI: 10.3389/fimmu.2021.773193
    The gastrointestinal immune system plays an important role in immune homeostasis regulation. It regulates the symbiotic host-microbiome interactions by training and developing the host's innate and adaptive immunity. This interaction plays a vital role in host defence mechanisms and at the same time, balancing the endogenous perturbations of the host immune homeostasis. The fish gastrointestinal immune system is armed with intricate diffused gut-associated lymphoid tissues (GALTs) that establish tolerance toward the enormous commensal gut microbiome while preserving immune responses against the intrusion of enteric pathogens. A comprehensive understanding of the intestinal immune system is a prerequisite for developing an oral vaccine and immunostimulants in aquaculture, particularly in cultured fish species. In this review, we outline the remarkable features of gut immunity and the essential components of gut-associated lymphoid tissue. The mechanistic principles underlying the antigen absorption and uptake through the intestinal epithelial, and the subsequent immune activation through a series of molecular events are reviewed. The emphasis is on the significance of gut immunity in oral administration of immunoprophylactics, and the different potential adjuvants that circumvent intestinal immune tolerance. Comprehension of the intestinal immune system is pivotal for developing effective fish vaccines that can be delivered orally, which is less labour-intensive and could improve fish health and facilitate disease management in the aquaculture industry.
  2. Liu Y, Andin VC, Chor WK, Gunasekaran B, Chong CM, Lee PT, et al.
    J Fish Biol, 2024 Aug 22.
    PMID: 39175254 DOI: 10.1111/jfb.15908
    This study aims to develop an alternative aquafeed derived from insect meal for Lates calcarifer juveniles, with the objectives of exploring the physiological performance, biological parameters, and economic analysis of substituting fishmeal (FM) with defatted black soldier fly (Hermetia illucens) larvae (BSFL) as part of the diet of L. calcarifer juveniles. Five practical diets were formulated to include 0% (BSFL0, serves as control group), 5% (BSFL5), 10% (BSFL10), 15% (BSFL15), and 20% (BSFL20) of BSFL meal, partially or fully replacing FM, respectively. Each diet was randomly assigned to triplicate groups of 30 fish (10.70 ± 0.07 g) per tank (300 L). The fish were fed twice daily to apparent satiation. A 56-day feeding trial was conducted to evaluate the impacts of defatted BSFL meal replacing FM on the growth performance, feed efficiency, composition analysis of fish muscle, cumulative mortality rate challenged with Vibrio parahaemolyticus, and economic returns of L. calcarifer. These results show that differences in weight gain and specific growth rate among the different treatments were statistically significant (p  BSFL0 > BSFL5 > BSFL15 > BSFL20. However, the feed conversion ratio and protein efficiency ratio showed the opposite trend as above. Although the diets experienced a decline in crude protein content and an increase in crude fat content with the increasing proportion of BSFL substituting FM, the crude protein and fat content of fish muscle were only slightly influenced. It is worth mentioning that levels of nonessential amino acids, delicious amino acids, saturated fatty acids, omega-6, omega-9 in BSFL10 group all showed an increase compared with the control group. After a 7-day challenge test with V. parahaemolyticus, the cumulative mortality rates of the BSFL5 and BSFL10 groups, respectively, dropped to 5.20%, 5.28% compared to the control group's 16.88%; however, the mortality rates of BSFL15 (34.67%) and BSFL20 (41.77%) groups were found to be significantly (p 15%) leads to a negative effect. From an economic point of view, 10% inclusion of BSFL in practical diets is recommended for L. calcarifer juveniles.
  3. Trung NB, Nan FH, Lee MC, Loh JY, Gong HY, Lu MW, et al.
    Fish Shellfish Immunol, 2021 Dec;119:587-601.
    PMID: 34743023 DOI: 10.1016/j.fsi.2021.11.001
    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.
  4. Vakaloloma U, Ho TH, Loh JY, Chong CM, Wangkahart E, Lee MC, et al.
    Vet Res Commun, 2023 Dec;47(4):1973-1990.
    PMID: 37349590 DOI: 10.1007/s11259-023-10152-8
    Rachycentron canadum (cobia) is a marine fish species of high economic value in aquaculture due to its fast growth rate and good feed conversion efficacy. Regrettably, the industry has been affected by significant setbacks from high mortality due to diseases. Consequently, an improved perception of innate immunity correlated to each mucosal-associated lymphoid tissue (MALT) in teleost fish is necessary to understand hosts' response towards infections better. The utilization of polysaccharides in seaweed to stimulate the immune system has gathered unprecedented attention. The present study examined the immunostimulatory effects of Sarcodia suae water extracts (SSWE) on in vivo gill-, gut- and skin-associated lymphoid tissues (GIALT, GALT, and SALT) via immersion and oral ingestions. The GIALT genes (TNF-α, Cox2, IL-1β, IL-6, IL-8, IL-17 A/F1-3, IL-11, IL-12, IL-15, IL-18, MHCIa, IgM, and IgT) except IL-10 recorded positive upregulations in a dose-dependent manner post 24 h immersion in SSWE, indicating the algae extract contained bioactive compounds that could stimulate the immune genes. The upregulation of IL-12, IL-15, and IL-18 in the gills and hindgut post-SSWE immersion indicated that the extract could promote Th1-related responses in the MALTs. The modulation of immune gene expressions in the feeding trial was less potent than in the SSWE immersion. These findings indicated that the SSWE stimulated robust immune responses in both the GIALT and GALT of cobia. This suggests that the SSWE could be further explored as an effective immersive stimulant for fish, enhancing their immune system against pathogens.
  5. Tran HTQ, Ho TH, Nan FH, Liu CH, Hu YF, Chong CM, et al.
    Fish Shellfish Immunol, 2024 May 01;150:109597.
    PMID: 38697373 DOI: 10.1016/j.fsi.2024.109597
    This study investigated the effects of fish protein hydrolysate derived from barramundi on growth performance, muscle composition, immune response, disease resistance, histology and gene expression in white shrimp (Penaeus vannamei). In vitro studies demonstrated FPH enhanced mRNA expressions of key immune-related genes and stimulated reactive oxygen species (ROS) production and phagocytic activity in shrimp hemocytes. To evaluate the effects of substituting fish meal with FPH in vivo, four isoproteic (43 %), isolipidic (6 %), and isoenergetic diets (489 kcal/100 g) were formulated with fish meal substitution levels of 0 % (control), 30 % (FPH30), 65 % (FPH65), and 100 % (FPH100). After 8-week feeding, the growth performance of FPH65 and FPH100 were significantly lower than that of control and FPH30 (p  0.05). The expressions of immune-related genes were analyzed at week 4 before and after challenge. In control group, V. parahaemolyticus significantly elevated SOD in hepatopancreas and Muc 19, trypsin, Midline-fas, and GPx in foregut (p  0.05). These findings suggest that FPH30 holds promise as a feed without adverse impacts on growth performance while enhancing the immunological response of white shrimp.
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