Department of Aquaculture Technology

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    The Current Status of Antibiotic-Resistant Bacteria and Resistance Genes in African Aquaculture
    (Springer, 2023-02-28) Samwel Mchele Limbu
    Aquaculture is currently the fastest-growing food production sector, supplying half of the fish consumed by humans globally. Nevertheless, aquaculture has been indicated as a medium for production and gateway for the transfer of antibiotic resistance bacteria (ARB) and their corresponding antibiotic resistance genes (ARGs). However, the precise sources, factors responsible, types and consequences of ARB and ARGs in African aquaculture are currently not well known. This chapter fills such as a knowledge gap by discussing the sources of ARB and ARGs in African aquaculture, the possible factors contributing to their increase, the types of ARB and ARGs found in the industry and the socioeconomic consequences to humans. The chapter found that, African aquaculture represents a hotspot reservoir of ARB and ARGs. However, the ARB and ARGs in African aquaculture are complex and not solely derived from the industry. They originate from multiple sources, including antibiotics application in aquaculture, livestock manure and integrated aquaculture systems, agriculture and human health. The ARB and ARGs also originate from wastewater treatment plants, hospitals and pharmaceutical industries. The African aquaculture sector is currently dominated by multiple ARB and their genes, resistant to antibiotics commonly used to treat human diseases. The ARB and ARGs in aquaculture are caused by overuse and misuse of antibiotics in the sector, lack of regulations, ineffective enforcement, loose rules and inadequate surveillance and monitoring systems, which increase the availability and accessibility of antibiotics. Moreover, incorrect prescription, easy affordability and prolonged use of the same antibiotics also contribute to the increased ARB and ARGs in the African aquaculture industry. The ARB and ARGs existing in aquaculture are transferred to humans via aquaculture product contamination during handling and consumption of raw or undercooked food, horizontal and vertical gene transfers and direct contact with water or drinking contaminated water. The consequences of ARB and ARGs from aquaculture include increased infections, frequency of treatment failures, severe infections, prolonged duration of illness and increased frequency of bloodstream infections and hospitalisation. These make primary healthcare redundant for previously curable diseases, thereby increasing costs to society on treating cultured species and humans, leading to animal and human deaths. The information generated in the chapter calls for a “One Health” approach to combat ARB and ARGs while formulating stringent measures, legislations and regulations for antibiotics use in aquaculture, agriculture and sick patients to safeguard human health.
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    Dietary docosahexaenoic acid reduces fat deposition and alleviates liver damage induced by D-galactosamine and lipopolysaccharides in Nile tilapia (Oreochromis niloticus)
    (Elsevier, 2023-06) Yi-Chan Liu; Samwel Mchele Limbu; Jin-Gang Wang; Mai Wang; Li-Qiao Chen; Fang Qiao; Yuan Luo; Mei-Ling Zhang; Zhen-Yu Du
    Liver health is important to maintain survival and growth of fish. Currently, the role of dietary docosahexaenoic acid (DHA) in improving fish liver health is largely unknown. This study investigated the role of DHA supplementation in fat deposition and liver damage caused by D-galactosamine (D-GalN) and lipopolysaccharides (LPS) in Nile tilapia (Oreochromis niloticus). Four diets were formulated as control diet (Con), Con supplemented with 1 % DHA, 2 % DHA and 4 % DHA diets, respectively. The diets were fed to 25 Nile tilapia (2.0 ± 0.1 g, average initial weight) in triplicates for four weeks. After the four weeks, 20 fish in each treatment were randomly selected and injected with a mixture of 500 mg D-GalN and 10 μL LPS per mL to induce acute liver injury. The results showed that the Nile tilapia fed on DHA diets decreased visceral somatic index, liver lipid content and serum and liver triglyceride concentrations than those fed on the Con diet. Moreover, after D-GalN/LPS injection, the fish fed on DHA diets decreased alanine aminotransferase and aspartate transaminase activities in the serum. The results of liver qPCR and transcriptomics assays together showed that the DHA diets feeding improved liver health by downregulating the expression of the genes related to toll-like receptor 4 (TLR4) signaling pathway, inflammation and apoptosis. This study indicates that DHA supplementation in Nile tilapia alleviates the liver damage caused by D-GalN/LPS through increasing lipid catabolism, decreasing lipogenesis, TLR4 signaling pathway, inflammation, and apoptosis. Our study provides novel knowledge on the role of DHA in improving liver health in cultured aquatic animals for sustainable aquaculture.
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    High cholesterol intake remodels cholesterol turnover and energy homeostasis in Nile tilapia (Oreochromis niloticus)
    (2023-02-16) Rui-Xin Li; Ling-Yun Chen; Samwel Mchele Limbu; Yu-Cheng Qian; Wen-Hao Zhou; Li-Qiao Chen; Yuan Luo; Fang Qiao; Mei-Ling Zhang
    · · ·The roles of dietary cholesterol in fish physiology are currently contradictory. The issue reflects the limited studies on the metabolic consequences of cholesterol intake in fish. The present study investigated the metabolic responses to high cholesterol intake in Nile tilapia (Oreochromis niloticus), which were fed with four cholesterol-contained diets (0.8, 1.6, 2.4 and 3.2%) and a control diet for eight weeks. All fish-fed cholesterol diets showed increased body weight, but accumulated cholesterol (the peak level was in the 1.6% cholesterol group). Then, we selected 1.6% cholesterol and control diets for further analysis. The high cholesterol diet impaired liver function and reduced mitochondria number in fish. Furthermore, high cholesterol intake triggered protective adaptation via (1) inhibiting endogenous cholesterol synthesis, (2) elevating the expression of genes related to cholesterol esterification and efflux, and (3) promoting chenodeoxycholic acid synthesis and efflux. Accordingly, high cholesterol intake reshaped the fish gut microbiome by increasing the abundance of Lactobacillus spp. and Mycobacterium spp., both of which are involved in cholesterol and/or bile acids catabolism. Moreover, high cholesterol intake inhibited lipid catabolic activities through mitochondrial β-oxidation, and lysosome-mediated lipophagy, and depressed insulin signaling sensitivity. Protein catabolism was elevated as a compulsory response to maintain energy homeostasis. Therefore, although high cholesterol intake promoted growth, it led to metabolic disorders in fish. For the first time, this study provides evidence for the systemic metabolic response to high cholesterol intake in fish. This knowledge contributes to an understanding of the metabolic syndromes caused by high cholesterol intake or deposition in fish.
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    Vitellogenin 1 is essential for fish reproduction by transporting DHA-contained phosphatidylcholine from liver to ovary
    (Elsevier, 2023-04) Sheng-Xiang Sun; Yi-Chan Liu; Samwel Mchele Limbu; Dong-Liang Li; Li-Qiao Chen; Mei-Ling Zhang; Zhan Yin; Zhen-Yu Du
    Vitellogenins (Vtgs) are essential for female reproduction in oviparous animals, yet the exact roles and mechanisms remain unknown. In the present study, we knocked out vtg1, which is the most abundant Vtg in zebrafish, Danio rerio via the CRISPR/Cas 9 technology. We aimed to identify the roles of Vtg1 and related mechanisms in reproduction and development. We found that, the Vtg1-deficient female zebrafish reduced gonadosomatic index, egg production, yolk granules and mature follicles in ovary compared to the wide type (WT). Moreover, the Vtg1-deficient zebrafish diminished hatching rates, cumulative survival rate, swimming capacity and food intake, but increased malformation rate, and delayed swim bladder development during embryo and early-larval phases. The Vtg1-deficiency in female broodstock inhibited docosahexaenoic acid-enriched phosphatidylcholine (DHA-PC) transportation from liver to ovary, which lowered DHA-PC content in ovary and offspring during larval stage. However, the Vtg1-deficient zebrafish increased gradually the total DHA-PC content via exogeneous food intake, and the differences in swimming capacity and food intake returned to normal as they matured. Furthermore, supplementing Vtg1-deficient zebrafish with dietary PC and DHA partly ameliorated the impaired female reproductive capacity and larval development during early phases. This study indicates that, DHA and PC carried by Vtg1 are crucial for female fecundity, and affect embryo and larval development through maternal-nutrition effects. This is the first study elucidating the nutrient and physiological functions of Vtg1 and the underlying biochemical mechanisms in fish reproduction and development.
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    Dietary β-conglycinin induces intestinal enteritis and affects glycerophospholipid and arginine metabolism in mirror carp (Cyprinus carpio)
    (Elsevier, 2023-03-30) Qiaohua Luo; Zuliang Zhou; Jianhua Zhao; Hong Xu; Samwel Mchele Limbu; Qiyou Xu
    The use of soybean meal in aquafeed industry is partially limited by anti-nutritional factors such as β-conglycinin (β-CG). However, the effects of β-CG on mirror carp (Cyprinus carpio) growth performance and intestinal health has not been elucidated. This study investigated the effects of β-CG levels on growth performance, intestine immunity, and intestinal lipid and protein metabolism in mirror carp. Four hundred and fifty mirror carp (4.22 ± 0.03) g were fed a control (CON) diet containing fish meal as the protein source and four experimental diets containing β-CG at 40 (β-CG1), 80 (β-CG2), 120 (β-CG3), and 160 g/kg (β-CG4) replacing the fishmeal, respectively. Each treatment had 30 fish replicated three times and the feeding was done for six weeks. The results showed that weight gain, specific growth rate, survival rate, protein efficiency, feed conversion ratio, and condition factor were not affected by dietary β-CG (P > 0.05). Interestingly, the fish fed on the β-CG diets upregulated significantly the expression of tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β) and eIF4E-binding protein (4E-BP) in the proximal intestine (PI) than those fed on the CON diet (P < 0.05). The fish fed on the β-CG2 diet decreased significantly the expression of transforming growth factor beta 1 (TGFβ1) in the PI compared to those fed on the CON diet. The fish fed on the β-CG diets downregulated significantly the expression of target of rapamycin (TOR) in the PI than those fed on the CON diet. The fish fed on β-CG diets downregulated significantly the expression of AMP-activated protein kinase alpha (AMPK), acetyl-CoA carboxylase (ACC), TOR in the mid intestine (MI) while, claudin11 gene was significantly increased (P < 0.05). In the distal intestine (DI), feeding the fish on the β-CG diets downregulated significantly the expression of AMPK and ACC genes, while increased the expression TNF-α, IL-1β and claudin11 (P < 0.05). Feeding the fish with the β-CG diets mostly altered glycerophospholipid metabolism and arginine biosynthesis in intestinal mucosa. These results suggest that dietary β-CG induces enteritis in different intestinal segments and changes glycerophospholipid metabolism and arginine biosynthesis pathways. Fish farmers using soybean meal in the diets should process the ingredients to reduce β-CG concentration before use.
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    Dietary sodium acetate improves high-fat diet utilization through promoting differential nutrients metabolism between liver and muscle in Nile tilapia (Oreochromis niloticus)
    (Elsevier, 2023-02-25) Wen-Hao Zhou; Samwel Mchele Limbu; Rui-Xin Li; Yuan Luo; Jiong Ren; Fang Qiao; Mei-Ling Zhang; Zhen-Yu Du
    igh-fat diet (HFD) often causes many negative effects and impairs fish growth. Short-chain fatty acids (SCFAs) such as acetates modify metabolic disorders and regulate body homeostasis. However, the effects of sodium acetate on alleviating HFD in fish is currently unknown. This study investigated the role of using dietary sodium acetate to alleviate adverse effects caused by HFD in fish. Three replicates (thirty fish each) of 4.8 ± 0.30 g Nile tilapia (Oreochromis niloticus) were fed with control diet (Con), high-fat diet (HFD) or HFD containing sodium acetate diet (HFD + NaAc) for eight weeks. After the feeding trial, Nile tilapia fed with HFD increased significantly tissue lipid deposition, reduced insulin sensitivity and suppressed glucose and lipid metabolism in both liver and muscle, accompanied with signs of metabolic disorders and liver damage. Moreover, HFD feeding inhibited muscle protein synthesis and impaired fish growth performance. However, the fish fed on HFD + NaAc improved significantly oxidative stress, inflammation, apoptosis and injury in liver compared to those fed on HFD. More importantly, dietary sodium acetate supplementation enhanced insulin sensitivity, promoted glucose catabolic utilization and protein synthesis in muscle through activation of insulin and mTOR signaling pathways, respectively, and markedly improved the growth performance. In contrast, dietary sodium acetate promoted hepatic pentose phosphate pathway, hepatic glycogen accumulation, and activated lipid catabolism to alleviate hepatic lipid deposition. Our study illustrates that sodium acetate alleviates differentially the adverse effects induced by feeding Nile tilapia with HFD in muscle and liver. SCFAs such as acetate may be used for improving HFD utilization in fish nutrition.
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    Atorvastatin remodels lipid distribution between liver and adipose tissues through blocking lipoprotein efflux in fish
    (American Physiological Society, 2023-03-01) Rui-Xin Li; Ling-Yun Chen; Samwel Mchele Limbu; Bing Yao; Yi-Fan Qian; Wen-Hao Zhou; Li-Qiao Chen; Fang Qiao; Mei-Ling Zhang; Zhen-Yu Du; Yuan Luo
    The regulation of cholesterol metabolism in fish is still unclear. Statins play important roles in promoting cholesterol metabolism development in mammals. However, studies on the role of statins in cholesterol metabolism in fish are currently limited. The present study evaluated the effects of statins on cholesterol metabolism in fish. Nile tilapia (Oreochromis niloticus) were fed on control diets supplemented with three atorvastatin levels (0, 12, and 24 mg/kg diet, ATV0, ATV12, and ATV24, respectively) for 4 wk. Intriguingly, the results showed that both atorvastatin treatments increased hepatic cholesterol and triglyceride contents mainly through inhibiting bile acid synthesis and efflux, and compensatorily enhancing cholesterol synthesis in fish liver (P < 0.05). Moreover, atorvastatin treatment significantly inhibited hepatic very-low-density lipoprotein (VLDL) assembly and thus decreased serum VLDL content (P < 0.05). However, fish treated with atorvastatin significantly reduced cholesterol and triglycerides contents in adipose tissue (P < 0.05). Further molecular analysis showed that atorvastatin treatment promoted cholesterol synthesis and lipogenesis pathways, but inhibited lipid catabolism and low-density lipoprotein (LDL) uptake in the adipose tissue of fish (P < 0.05). In general, atorvastatin induced the remodeling of lipid distribution between liver and adipose tissues through blocking VLDL efflux from the liver to adipose tissue of fish. Our results provide a novel regulatory pattern of cholesterol metabolism response caused by atorvastatin in fish, which is distinct from mammals: cholesterol inhibition by atorvastatin activates hepatic cholesterol synthesis and inhibits its efflux to maintain cholesterol homeostasis, consequently reduces cholesterol storage in fish adipose tissue.
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    Dietary magnesium improves growth, antioxidant capacity, ammonia tolerance and intestinal health of juvenile freshwater prawn, Macrobrachium nipponense
    (Elsevier, 2022-11-15) Youqin Kong; Zhili Ding; Shanshan Wei; Changle Qi; Yan Liu; Yixiang Zhang; Samwel Mchele Limbu; Jinyun Ye
    The present study evaluated the effects of magnesium (Mg) on growth, antioxidant capacity, intestinal morphology and ammonia nitrogen resistance in juvenile oriental river prawn, Macrobrachium nipponense. Six semipurified diets (crude protein 401.1 g kg−1 diet; crude lipid 88.7 g kg−1 diet) containing Mg levels of 1.1, 1.4, 1.6, 2.1, 2.8 or 4.5 g kg−1 were formulated. A total of 1080 healthy juvenile prawns (0.151 ± 0.003 g) were randomly stocked into eighteen 300 L tanks (100 × 80 × 60 cm), each tank with 60 prawns. Each experimental diet was fed to prawns in triplicate twice daily (08:00 and 17:00) for eight weeks. After the feeding trial, 20 prawns from each tank were challenged with 37 mg/L ammonia nitrogen for 96 h, and the hemolymph, hepatopancreas and muscle of the rest of the prawns were collected. Final biomass was higher in prawns fed diets containing 1.6 and 2.1 g Mg kg−1 compared with those fed the diet with 1.1 g kg−1. Higher alkaline phosphatase (ALP) activity was obtained in the serum of prawns fed the 2.1 and 2.8 g Mg kg−1 diets. The highest activities of hepatopancreas superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and total antioxidant capacity (T-AOC) were observed in prawns fed 1.6–2.8 g Mg kg−1 of feed, and the lowest hepatopancreas malondialdehyde (MDA) concentrations and serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities were found in these treatment groups. Similarly, the prawns fed diets with 1.6 and 2.1 g Mg kg−1 had significantly lower MDA concentrations and cumulative mortality and higher antioxidant enzyme activities after ammonia nitrogen exposure. The intestinal epithelial cells were closely attached to the membrane in prawns fed the 2.1 g Mg kg−1 diet, but they were partly detached in those fed the 1.1 and 4.5 g Mg kg−1 diets. The minimum dietary Mg supplementations of juvenile M. nipponense were determined to be 1.6, 1.9, 2.0 and 2.2 g kg−1 feed, respectively, using broken-line regression analysis based on the final biomass, SOD, ALT and ALP.
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    Inhibition of pyruvate dehydrogenase kinase increases carbohydrate utilization in Nile tilapia by regulating PDK2/4-PDHE1α axis and insulin sensitivity
    (Elsevier, 2022-06-24) Yuan Luo; Wen-Hao Zhou; Rui-Xin Li; Samwel Mchele Limbu; Fang Qiao; Li-Qiao Chen; Mei-Ling Zhang; Zhen-Yu Du
    Pyruvate dehydrogenase kinases (PDKs)-pyruvate dehydrogenase E1α subunit (PDHE1α) axis plays an important role in regulating glucose metabolism in mammals. However, the regulatory function of PDKs-PDHE1α axis in the glucose metabolism of fish is not well known. This study determined whether PDKs inhibition could enhance PDHE1α activity, and improve glucose catabolism in fish. Nile tilapia fingerlings (1.90 ± 0.11 g) were randomly divided into 4 treatments in triplicate (30 fish each) and fed with control diet without dichloroacetate (DCA0) (38% protein, 7% lipid and 45% corn starch) and the control diet supplemented with DCA, which inhibits PDKs through binding the allosteric sites, at 3.75 (DCA3.75), 7.50 (DCA7.50) and 11.25 g/kg (DCA11.25), for 6 wk. The results showed that DCA3.75, DCA7.50 and DCA11.25 significantly increased weight gain, carcass ratio and protein efficiency ratio (P < 0.05) and reduced feed efficiency (P < 0.05) of Nile tilapia. To investigate the effects of DCA on growth performance of Nile tilapia, we selected the lowest doseDCA3.75 for subsequent analysis. Nile tilapia fed on DCA3.75 significantly reduced the mesenteric fat index, serum and liver triglyceride concentration and total lipid content in whole fish, and down-regulated the expressions of genes related to lipogenesis (P < 0.05) compared to the control. The DCA3.75 treatment significantly improved glucose oxidative catabolism and glycogen synthesis in the liver, but significantly reduced the conversion of glucose to lipid (P < 0.05). Furthermore, the DCA3.75 treatment significantly decreased the PDK2/4 gene and protein expressions (P < 0.05), accordingly stimulated PDHE1α activity by decreasing the phosphorylated PDHE1α protein level. In addition, DCA3.75 treatment significantly increased the phosphorylated levels of key proteins involved in insulin signaling pathway and glycogen synthase kinase 3β (P < 0.05). Taken together, the present study demonstrates that PDK2/4 inhibition by using DCA promotes glucose utilization in Nile tilapia by activating PDHE1α, and improving insulin sensitivity. Our study helps to understand the regulatory mechanism of glucose metabolism for improving dietary carbohydrate utilization in farmed fish.
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    Black soldier fly (Hermetia illucens, L.) larvae improves growth performance, feed efficiency and economic returns of Nile tilapia (Oreochromis niloticus, L.) fry
    (Wiley, 2022-05-23) Samwel Mchele Limbu; Amon Paul Shoko; Eusebia Ernest Ulotu; Siwema Amran Luvanga; Fridah Mukiri Munyi; John Obedy John; Mary Adhiambo Opiyo
    Insects have been proposed as potential alternative animal protein sources to replace fishmeal (FM), which is expensive and has limited availability for fish feed formulation. However, studies on the effects of replacing FM with black soldier fly (Hermetia illucens) larvae (BSF-L) on growth performance of Nile tilapia (Oreochromis niloticus) fry, water quality and economic benefits are currently limited. This study determined the effects of replacing 100% FM, 75% FM, 50% FM, 25% FM and 0% FM with cheap BSF-L meal, hereafter referred to as BSF-L0, BSF-L25, BSF-L50, BSF-L75 and BSF-L100 diets, respectively for rearing all-male Nile tilapia fry produced by YY technology. The study further determined the optimum percentage of BSF-L meal for maximum growth of Nile tilapia fry. A total of 2400 visually healthy Nile tilapia fry weighing 0.001 g were randomly stocked into 20 plastic tanks (120 fry per tank, four replicates per treatment) and fed the BSF-L diets for 12 weeks. The BSF-L75 diet increased significantly specific growth rate, total weight gain, Zihler's index of fry and nitrate in the culture water but reduced feed conversion ratio of diet and total suspended solids in the culture water (p < 0.05). Feeding the Nile tilapia fry with BSF-L50 diet increased significantly hepatosomatic index (p < 0.05). The diets with BSF-L75 and BSF-L100 reduced significantly the incidence cost by 31.97% and 28.77% (p < 0.05), and increased profit index by 3.97 and 3.44%, respectively. The optimum percentage of BSF-L inclusion required for maximum growth performance of Nile tilapia fry was estimated as 81% to 84% based on polynomial analysis. Taken together, feeding Nile tilapia fry with diets containing 81% to 84% BSF-L meal improved growth performance. The BSF-L75 diet enhanced feed efficiency and had no deleterious effect on the liver and intestines. Meanwhile, it improved nitrate concentration for increased natural productivity. Incorporating 75% BSF-L meal in diets for Nile tilapia fry reduced 30% of feed cost leading to 4% higher economic returns.
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    The transforming growth factor beta (TGF-β/Smads) pathway regulates collagen synthesis and deposition in swim bladder of Chu's croaker (Nibea coibor) stimulated by proline
    (2022-09-15) Hua Rong; Fan Lin; Lijun Ning; Kun Wu; Samwel Mchele Limbu; Qingchao Shi; Chuanjie Qin; Xiaobo Wen
    The swim bladder performs various functions in fish, is consumed by humans and can be processed into high-grade fish glue (traditional high-value tonic). However, little is known on the optimum amount of proline (Pro) required for promoting collagen synthesis and deposition in fish swim bladder and the regulatory mechanism. This study explored the effects and optimum amount of dietary Pro supplementation on collagen synthesis and deposition in Chu's croaker (Nibea coibor) swim bladder and its possible molecular mechanism. A total of 450 juvenile fish (8.64 ± 0.14 g) were randomly stocked into 18 cages (25 fish per cage) in triplicate and fed Pro supplemented diets (0, 5, 10, 15, 20 and 25 g kg−1) for eight weeks. Another 225 fish (11.62 ± 0.15 g) were distributed randomly into control, Pro and Pro diet plus injected with specific inhibitor of Smad3 (SIS3) in order to inhibit the transforming growth factor beta (TGF-β)/Smads pathway. Pro supplementation up-regulated significantly the expression of genes involved in TGF-β/Smads pathway such as collagen, type I, alpha 1 (col1a1) and 2 (col1a2), TGF-β and Smad2/3, and increased collagen content in the Chu's croaker swim bladder (P < 0.05). The optimum dietary Pro supplementation for deposition of collagen in Chu's croaker swim bladder was 19.36 g kg−1. The collagen content in the swim bladder positively correlated significantly with the mRNA expression of col1a1, col1a2, TGF-β and Smad2/3 genes (P < 0.05). The optimum dietary Pro supplementation for maximum synthesis of collagen in Chu's croaker swim bladder was 13.25, 13.32 and 15.94 g kg−1 based on mRNA expression of Smads2, Smads3 and col1a2 genes, respectively. SIS3 down-regulated the expression of col1a1, col1a2, TGF-β and Smad2/3 genes, subsequently decreased collagen deposition in the swim bladder (P < 0.05). These results suggest that 13.25 to 19.36 g kg−1 Pro supplementation improve collagen synthesis and deposition in Chu's croaker swim bladder. The TGF-β/Smad signaling pathway regulates collagen synthesis and deposition in Chu's croaker swim bladder. Our results provide an understanding on the molecular mechanism of collagen synthesis and deposition in fish for producing collagen required for human food and fish glue.
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    Dietary L-carnitine alleviates the adverse effects caused by reducing protein and increasing fat contents in juvenile largemouth bass (Micropterus salmoides)
    (2022-05-06) Yi-Chan Liu; Samwel Mchele Limbu; Jin-Gang Wang; Jiong Ren; Fang Qiao; Mei-Ling Zhang; Zhen-Yu Du
    Protein ingredients for formulation of fish feeds are expensive and have limited availability. Therefore, reducing dietary protein while increasing dietary fat content is a common practice in rearing carnivorous fish species. However, the ability of dietary L-carnitine to alleviate adverse effects in such diets is currently unknown. This study investigated the role of L-carnitine supplementation in alleviating adverse effects on growth performance, energy metabolism, antioxidant capacity, and inflammation response in juvenile largemouth bass (Micropterus salmoides) fed on a low protein and high fat diet. Three diets were formulated to contain low protein and high fat (LPHF: 420 g kg-1 protein and 150 g kg-1 lipid), LPHF supplemented with L-carnitine (LPHFC: 420 g kg-1 protein and 150 g kg-1 lipid), and a control diet (CON: 480 g kg-1 protein and 130 g kg-1 lipid). The diets were fed to 30 largemouth bass (g) juveniles in triplicates for eight weeks. The results showed that the fish feed on LPHF diet increased hepatosomatic index, visceral somatic index, mesenteric fat index, whole-body crude fat content, serum and liver triglyceride concentrations, and serum non-esterified fatty acid level than those fed on CON diet. Moreover, the fish fed on LPHF diet increased serum alanine aminotransferase activity and liver malondialdehyde content and reduced superoxide dismutase (SOD) activities in the serum and liver. Furthermore, the fish fed on LPHF diet reduced the whole-body crude protein content. Interestingly, feeding the fish on the LPHFC diet decreased fat deposition and liver damage by downregulating the expression of genes related to lipogenesis, inflammation, and increased SOD activity. This study indicates that L-carnitine supplementation in largemouth bass alleviates the adverse effects caused by LPHF diet by decreasing lipogenesis and increasing lipid catabolism. Our study provides novel knowledge on strategies to improve utilization of LPHF diet in cultured aquatic animals.
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    Cloning, tissue distribution and mRNA expression analysis of typeIcollagen alpha 1 gene from Chu's croaker (Nibea coibor)
    (Elsevier, 2022-05-25) Hua Rong; Fan Lin; Lijun Ning; Kun Wu; Baojia Chen; Jia Zheng; Samwel Mchele Limbu; Xiaobo Wen
    The demand for collagen has been increasing over years due to its wide application in food, cosmetics and biomedicine industries. The synthesis of collagen protein in fish depends on instructions provided by collagen, type I, alpha 1 (COL1A1) gene. However, cloning, tissue distribution and mRNA expression of COL1A1 gene in a gel-producing Chu's croaker (Nibea coibor) is currently unknown. This study cloned the cDNA of COL1A1 gene (GenBank accession number: MK641512) from six N. coibor fish. The distribution and mRNA expression pattern of COL1A1 was analyzed in eight tissues of N. coibor. The COL1A1 cDNA had a full length of 6130 bp and contained a 4344 bp open reading frame (ORF) encoding a polypeptide of 1448 amino acids. The homology of N. coibor COL1A1 amino acid had 98% similarity with Larimichthys crocea, indicating conservatism with other members in same family (Sciaenidae). The deduced polypeptide contained the same signal peptides, C-propeptide and N-propeptide domains, and triple helix domains, which are the characteristics of type I collagen in vertebrates. The mRNA of COL1A1 gene was expressed significantly higher in the spine of N. coibor than in all other tissues (P < 0.05), followed by swim bladder, skin and scales. The swim bladder had higher collagen and hydroxyproline contents than other tissues, followed by spine >, scales > and > skin (P < 0.05). Our study successfully cloned the COL1A1 gene from N. coibor for the first time. The COL1A1 gene contained all the features of collagen pro-α1(I) chain proteins, and shared high homology with other marine teleost. COL1A1 gene in N. coibor is highly expressed in spine and swim bladder, consistent with collagen distribution. Our study contributes to better understanding on collagen biosynthesis in N. coibor tissues for various industrial uses.
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    Aflatoxin B1 variations in animal feeds along the supply chain in Tanzania and its possible reduction by heat treatment
    (Taylor & Francis, 2022-03-04) Anjelina W. Mwakosya; Samwel Mchele Limbu; Nuria Majaliwa; Xiaobo Zou; Jiyong Shi; Oscar Kibazohi
    This study evaluated the presence of aflatoxin B1 in five different animal feeds collected from manufacturers, suppliers and consumers and its possible reduction by heating at 100°C for 180 min. A total of 160 animal feed samples were collected and analyzed by using lateral flow immunoassay method. The results revealed that all animal feeds analyzed were positive for aflatoxin B1 with 91% samples containing high concentrations ranging from 24.00 to 76.23 ng/g above the international allowable standard for animal feeds (20 ng/g). Maize bran (76 ng/g) and sunflower cake (63 ng/g) had higher aflatoxin B1 concentrations, correlating with higher moisture content. Upon heating the feeds, aflatoxin B1 was reduced to a concentration ranging from 2.24 to 9.78 ng/g (<20 ng/g). Our study suggests high potential health problems to animals and humans from aflatoxins requiring proper heating and frequent monitoring of the animal feeds for aflatoxin B1.
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    Dietary L-carnitine supplementation recovers the increased pH and hardness in fillets caused by high-fat diet in Nile tilapia (Oreochromis niloticus)
    (Elsevier, 2022-07-15) Zhi-Yong Zhang; Samwel Mchele Limbu; Si-Han Zhao; Li-Qiao Chen; Yuan Luo; Mei-Ling Zhang; Fang Qiao; Zhen-Yu Du
    The wide use of high-fat diet (HFD) causes negative effects on flesh quality in farmed fish. l-carnitine, a lipid-lowering additive, enhances mitochondrial fatty acid β-oxidation. However its roles in alleviating the effects of HFD on flesh quality in fish are unknown. We fed Nile tilapia with medium-fat diet (MFD, 6% dietary lipid), high-fat diet (HFD, 12% dietary lipid) and HFCD supplemented with l-carnitine (HFCD + 400 mg/kg l-carnitine) for 10 weeks. The HFD-fed fish had higher fat deposition, pH value, myofiber density and flesh hardness than those fed on MFD. However, feeding the fish with the HFCD improved lipid catabolism, which increased significantly lactic acid content and myofiber diameter in muscle, thus reduced pH and hardness values. HFCD also reduced endoplasmic reticulum stress and myofiber apoptosis caused by HFD in the fish. Our study suggests that dietary l-carnitine supplementation alleviates the negative effects of HFD on flesh quality of farmed fish.
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    Arginine supplementation in plant-rich diets affects growth, feed utilization, body composition, blood biochemical indices and gene expressions of the TOR signaling pathway in juvenile Asian red-tailed catfish (Hemibagrus wyckoiides)
    (Wiley, 2022-02) Dahai Gu; Jingying Zhao; Samwel Mchele Limbu; Yunan Liang; Junming Deng; Baoliang Bi; Lingfu Kong; Hui Yan; Xiaowen Wang; Qing Hu; Yu Gao; Dan Jia; Hua Rong
    Arginine (Arg) is an important essential amino acid required for optimum fish growth. However, its role in plant-rich diets on growth and the required level in juvenile Hemibagrus wyckioides are currently unknown. This study evaluated the effects of Arg supplementation in plant-rich diets on growth, feed utilization, whole body composition, blood biochemical indicators and gene expressions of the target of rapamycin (TOR) signaling pathway in juvenile H. wyckioides. A total of 450 fish (6.40 ± 0.06 g) were randomized into six equal groups and fed on diets with graded Arg levels: 0 (control), 3, 6, 9, 12 and 15 g/kg of dry feed for eight weeks. The results revealed that specific growth rate (SGR) and feed conversion ratio of juvenile H. wyckioides were significantly improved and reduced, respectively in fish fed on 9 g/kg of Arg supplementation (p < .05). The optimum amount of dietary Arg content in plant-rich diets for H. wyckioides was 63.77 g/kg diet based on SGR. Feeding fish with dietary Arg supplementation ranging from 9 to 12 g/kg improved significantly (p < .05) whole body protein contents and decreased lipid deposition. The mRNA levels of TOR, eukaryotic translation initiation factor 4E (4EBP) and ribosomal protein S6 kinase 1 (S6K1) in muscle and liver were significantly affected by increasing dietary Arg levels. Meanwhile, the mRNA relative expression levels of TOR and S6K1 in the liver and muscle increased significantly as Arg supplementation increased. On the other hand, the mRNA relative expression levels of 4EBP was significantly higher in fish fed on the 9 g/kg Arg diet compared to those fed on the control and 15 g/kg Arg diets. In conclusion, our current results indicate that dietary Arg supplementation in plant-rich diets, might activate the TOR signaling pathway, which promotes protein synthesis and reduces lipid accumulation of H. wyckioides, resulting in improved growth.
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    Modulation of growth, antioxidant status, hepatopancreas morphology, and carbohydrate metabolism mediated by alpha-lipoic acid in juvenile freshwater prawns Macrobrachium nipponense under two dietary carbohydrate levels.
    (Elsevier, 2022-01-15) Ding Z; Xiong Y; Zheng J; Zhou D; Kong Y; Qi C; Liu Y; Ye J; Samwel Mchele Limbu
    Carbohydrate is commonly used in aquatic organism as the cheapest energy source. However, most aquatic animals are low efficient in utilizing dietary carbohydrate for energy provision. Therefore, improving the utilization efficiency of high carbohydrate by aquatic organisms is important for sustainable aquaculture development. Alpha-lipoic acid (α-LA) is a cofactor of key enzymes involved in glucose metabolism. However, its role in regulation of carbohydrate metabolism in aquatic animals is currently unknown. This study investigated the effects of α-LA on growth, antioxidant status, hepatopancreas morphology, and carbohydrate metabolism of freshwater prawns Macrobrachium nipponense under two dietary carbohydrate levels. Six diets were formulated using casein and fish meal as the protein sources, fish oil and soybean oil as the lipid sources, and corn starch as the carbohydrate source, at two carbohydrate levels (15% and 30%) and three concentrations of α-LA supplementation (0, 700, and 1400 mg/kg). Each experimental diet was fed to juvenile prawns (mean weight 0.050 ± 0.003 g), twice daily to apparent satiation, in five replicates. In the 15% (low)-carbohydrate groups, weight gain was significantly reduced in prawns fed 1400 mg/kg α-LA compared with 0 or 700 mg/kg α-LA, but there was no significant effect in the 30% (high)-carbohydrate groups. Survival was significantly higher among prawns fed 700 mg/kg α-LA compared with 0 or 1400 mg/kg α-LA regardless of the carbohydrate level. The levels of dietary carbohydrate and α-LA also significantly affected the activities of key enzymes involved in glycolysis and the citric acid cycle. The activities of hexokinase and pyruvate kinase activities were highest in prawns fed l400 mg/kg α-LA, in both the low- and high-carbohydrate groups. Supplementation with 700 or 1400 mg/kg α-LA significantly increased the activities of 6-phosphofructokinase, pyruvate dehydrogenase, and succinate dehydrogenase regardless of the carbohydrate level. The level of malondialdehyde decreased significantly with increased α-LA supplementation; in contrast, the levels of superoxide dismutase, glutathione peroxidase, and glutathione were significantly higher in prawns fed 700 or 1400 mg/kg α-LA when compare with those fed no α-LA supplement. Dietary α-LA did not affect the overall morphology of the hepatopancreas, but 1400 mg/kg α-LA reduced the number of B (blister-like) cells. Thus, 700 mg/kg α-LA supplementation under either dietary carbohydrate level could improve the antioxidant status, carbohydrate metabolism, and morphology of the hepatopancreas in juveniles of M. nipponense. We conclude that α-LA supplementation can be used to improve the efficiency of carbohydrate utilization when a 30% carbohydrate diet is used for this prawn.
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    More simple more worse: Simple carbohydrate diets cause alterations in glucose and lipid metabolism in Nile tilapia (Oreochromis niloticus).
    (Elsevier, 2022-03-15) Wen-Hao Zhou; Chen-Chen Wu; Samwel Mchele Limbu; Rui-Xin Li; Li-Qiao Chen; Fang Qiao; Yuan Luo; Mei-Ling Zhang; Tao Han; Zhen-Yu Du
    Dietary carbohydrates are widely used in aquafeeds as cheap energy source and improve quality of feeds. However, comprehensive assessment of the effects of dietary carbohydrate complexity on fish metabolism and liver health is currently lacking. This study investigated the effect of replacing 50% of starch by using sucrose, fructose and glucose. Four isonitrogenic (400 g/kg protein) and isolipidic (60 g/kg fat) feeds containing starch (polysaccharide), sucrose (disaccharide) and fructose and glucose (monosaccharides) were fed to three replicates of 4.5 ± 0.30 g Nile tilapia (Oreochromis niloticus) for eight weeks. Afterwards, growth performance, feed utilization efficiency, glucose and lipid metabolism and liver health were evaluated. The results showed that the Nile tilapia fed on monosaccharide diets had significantly lower growth and feed utilization efficiency than those fed on disaccharide and polysaccharide diets. The Nile tilapia fed on monosaccharide diets reduced significantly lipid deposition, hepatosomatic index (HSI), serum alanine transaminase (ALT) and aspartate aminotransferase (AST) and expression of lipid metabolism genes than those fed on polysaccharide diet. The Nile tilapia fed on disaccharide diet increased significantly the serum insulin, muscle glycogen, and muscle glycogen synthase (gs) gene expression compared with those fed on polysaccharide diet. Interestingly, the Nile tilapia fed on monosaccharide diets reduced significantly the glycogen content while they increased insulin, glucose and the expression of liver gluconeogenesis genes such as glucose-6-phosphatase (g6pase) and phosphoenolpyruvate carboxykinase (pepck) than those fed on polysaccharide diet. Contrary, the Nile tilapia fed on disaccharide diet downregulated liver glucose catabolism and lowered serum triglyceride (TG) levels than those fed on polysaccharide but enhanced muscle peroxisome proliferator activated receptor alpha (pparα) and carnitine palmitoyltransferase 1a (cpt1a) gene expression, liver gluconeogenesis and pentose phosphate pathways. These results indicate that Nile tilapia utilizes better dietary polysaccharides and disaccharides than monosaccharides. Feeding Nile tilapia on monosaccharide diets caused insulin resistance and glucose metabolism disorders manifested by hyperinsulinism and hyperglycemia, respectively. The carbohydrate complexity affects the nutritional metabolism and liver health of fish.
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    Lipolysis and lipophagy play individual and interactive roles in regulating triacylglycerol and cholesterol homeostasis and mitochondrial form in zebrafish
    (Elsevier, 2021-06-08) Si-Lan Han; Yu-Cheng Qian; Samwel Mchele Limbu; Jing Wang; Li-Qiao Chen; Mei-Ling Zhang; Zhen-Yu Du
    Neutral lipases-mediated lipolysis and acid lipases-moderated lipophagy are two main processes for degradation of lipid droplets (LDs). However, the individual and interactive roles of these metabolic pathways are not well known across vertebrates. This study explored the roles of lipolysis and lipophagy from the aspect of neutral and acid lipases in zebrafish. We established zebrafish strains deficient in either adipose triglyceride lipase (atgl−/−; AKO fish) or lysosomal acid lipase (lal−/−; LKO fish) respectively, and then inhibited lipolysis in the LKO fish and lipophagy in the AKO fish by feeding diets supplemented with the corresponding inhibitors Atglistatin and 3-Methyladenine, respectively. Both the AKO and LKO fish showed reduced growth, swimming activity, and oxygen consumption. The AKO fish did not show phenotypes in adipose tissue, but mainly accumulated triacylglycerol (TAG) in liver, also, they had large LDs in the hepatocytes, and did not stimulate lipophagy as a compensation response but maintained basal lipophagy. The LKO fish reduced total lipid accumulation in the body but had high cholesterol content in liver; also, they accumulated small LDs in the hepatocytes, and showed increased lipolysis, especially Atgl expression, as a compensatory mechanism. Simultaneous inhibition of lipolysis and lipophagy in zebrafish resulted in severe liver damage, with the potential to trigger mitophagy. Overall, our study illustrates that lipolysis and lipophagy perform individual and interactive roles in maintaining homeostasis of TAG and cholesterol metabolism. Furthermore, the interactive roles of lipolysis and lipophagy may be essential in regulating the functions and form of mitochondria.
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    Different effects of two dietary levels of tea polyphenols on the lipid deposition, immunity and antioxidant capacity of juvenile GIFT tilapia (Oreochromis niloticus) fed a high-fat diet. Aquaculture, 542: 736896. https://doi.org/10.1016/j.aquaculture.2021.736896
    (Elsevier, 2021-05-11) Yu-Cheng Qian; Xue Wang; Jiong Ren; Jie Wang; Samwel Mchele Limbu; Rui-Xin Li; Wen-Hao Zhou; Fang Qiao; Mei-Ling Zhang; Zhen-Yu Du
    Long-term feeding of fish with a high-fat diet (HFD) causes excess fat deposition and an impairment of immune function. In the present study, we aimed to determine whether dietary tea polyphenols (TPs) would ameliorate the adverse effects of HFD-feeding in GIFT tilapia. Juvenile GIFT tilapias (5.4 ± 0.9 g) were raised in twelve 200-L tanks (three tanks per diet, 20 fish per tank) and fed a control diet (6% fat, 36% protein), an HFD (12% fat, 36% protein), or an HFD supplemented with 50 mg/kg or 200 mg/kg TP for 8 weeks. The fish were hand-fed 5% of their body weight per day in three feeds, and maintained at 28 ± 1 °C under a 14-h light/10-h dark cycle. The fish in each tank were bulk weighed and counted fortnightly, and the daily feed amount was adjusted accordingly. At the end of the trial, the cumulative survival rate was calculated, and the weight gain and feed conversion ratio were calculated according to the bulk weight of fish in each tank. Tissues were collected from nine fish per diet, their organs were weighed, and biochemical and molecular indices were subsequently measured. HFD-feeding significantly increased lipid deposition, reduced cumulative survival from 96% to 75%, reduced hepatic alkaline phosphate activity (AKP) and serum total antioxidant capacity (T-AOC); and reduced the hepatic expression of immunoglobulin M (IgM), transforming growth factor-beta (TGF-β) and glutathione-S-transferase (GST) genes versus the control diet. The addition of TPs at 50 or 200 mg/kg both ameliorated the HFD-induced increase in lipid droplets in the liver (50 mg/kg TP from 40.83% to 17.27%; 200 mg/kg TP to 25.33%), and increased the cumulative survival rate of the tilapia. The addition of 50 mg/kg TP had a marked effect increasing cumulative survival to 90%, and increasing the activities of serum acid phosphatase (ACP), T-AOC; and IgM, TGF-β, nuclear factor-κB (NF-κB), superoxide dismutase (SOD), and GST gene expression to the highest level of the HFD-fed groups. The 50 mg/kg TP-containing diet also significantly increased the hepatic expression of carnitine palmitoyltransferase 1 alpha (CPT1α) versus the control diet. In contrast, the tilapia fed an HFD supplemented with 200 mg/kg TPs had the lowest expression of adipose triglyceride lipase, hormone-sensitive lipase, CPT1α, fatty acid synthase and acetyl-CoA carboxylase alpha genes of any of the groups, which implies that the lower and higher levels of TP supplementation have differing effects on lipid metabolism. The 200 mg/kg supplement had lower cumulative survival rate (82%), and smaller effects on serum ACP and hepatic AKP activities than the 50 mg/kg dose, and had no significant effect on serum T-AOC or the expression of IgM, TGF-β, GST, or NF-κB genes in the tilapia. These results indicate that the beneficial effects of TPs on the lipid metabolism and health of fish fed an HFD are dose-related. Moreover, they are likely to be largely mediated through lipid catabolism.