Effects of dietary hydrolysate supplementation on growth, body composition, hematological responses, and liver histology of juvenile giant trevally (Caranx ignobilis Forsskal, 1775).

A feeding study was conducted to investigate how fish protein hydrolysate (FPH) supplementation affected the growth, feed utilization, body composition, and hematology of juvenile giant trevally (Caranx ignobilis Forsskal, 1775). Seven isonitrogenous (52% protein) and isocaloric diets (10% lipid) were formulated, wherein shrimp hydrolysate (SH) and tuna hydrolysate (TH) were used to replace fishmeal at inclusion levels of 0 (control), 30, 60, and 90 g/kg and labeled as control, SH30, SH60, SH90, TH30, TH60, and TH90, respectively. Each diet was fed to triplicate groups of juvenile giant trevally for 8 weeks. The results showed higher final body weight and specific growth rate in fish fed SH30, SH60, TH30, and TH60 than fed control diet. No difference was observed in feed intake, but reduced feed conversion ratio (FCR) was found in fish fed SH30, SH60, TH30, and TH60, demonstrating these diets improved feed utilization. TH90 caused deposition of lipid droplet in the hepatocyte, a sign of liver damage. Total monounsaturated fatty acids, polyunsaturated fatty acids (PUFA), and highly unsaturated fatty acids in fish were not affected by FPH supplementation. Fish fed TH30 showed lower ∑n - 3 PUFA than the fish fed remaining dietary treatments. The elevated serum protein was seen in fish fed control, SH30, SH60, and TH30, demonstrating that these diets were beneficial for the innate immune response in giant trevally. The results indicate that TH and SH could be incorporated into diets of giant trevally at 30-60 g/kg, replacing 7%-13% fishmeal with enhanced growth and health benefits.

From waste to feed: Microbial fermented abalone waste improves the digestibility, gut health, and immunity in marron, Cherax cainii.

New aquafeed ingredients produced by a circular economy approach are the opportunity for sustainable and resilient aquaculture. In the light of this approach, the mixture of abalone waste and Sargassum spp (9:1) fermented by Saccharomyces cereviceae and Lactobacillus casei (Yakult®) (FMAS) were used to replace 0% (FMAS0), 25% (FMAS-25), 50% (FMAS-50), 75% (FMAS-75), and 100% (FMAS-100) of fishmeal (FM) protein in marron, Cherax cainii diet. The marron was fed these diets in triplicate for 90 days. Growth, feed utilization and protein efficiency ratio were unchanged in marron-fed all test diets. Improvement in apparent protein digestibility was aligned with an increase in the size and number of B-cells in the hepatopancreas. Most of the immune responses, except for haemocyte clotting time, hyaline cells and neutral red retention time (NRR time) were unchanged by 42- and 90-days feeding trials compared to those of the control group. 90 days post-feeding marron with FMAS25 showed a lower haemocyte clotting time than the post 42 days feeding marron with the same diet. Hyaline cells increased in marron fed FMAS75 for 90 days compared to marron fed the same diet for 42 days. The challenge test involved injecting marron with Vibrio mimicus resulted in a 100% survival rate after 96 h of exposure. During the challenge test, phagocytosis activity in 24 and 48-h post-challenged marron fed FMAS75 decreased which recovered after 96 h post-challenge. Marron fed FMAS50 also recorded a significantly higher proportion of granular cells after 24 h and NRR time at 96 h compared with that of other treatments. Given the above indicators of bio-growth, feed efficiency and immune responses, total replacement of FM protein of marron practical feed with FMAS are considered feasible and optimum to maintain health status and resistance to disease.

The Effect of Two Dietary Protein Sources on Water Quality and the Aquatic Microbial Communities in Marron (Cherax cainii) Culture.

Feeding freshwater crayfish species with different diets not only affects the water quality but also induces the abundance of various microbial communities in their digestive tracts. In this context, very limited research has been undertaken to understand the impacts of various protein incorporated aqua-diets on the characteristics of water and its microbial communities. In this study, we have critically analysed the water quality parameters including pH, dissolved oxygen, nitrate, nitrite, ammonia and phosphorus, as well as bacterial communities under marron (Cherax cainii) aquaculture, fed fishmeal (FM) and poultry by-product meal (PBM)-based diets for 60 days. The results unveiled that over the time, feeding has significant impacts on organic waste accumulation, especially ammonia, nitrate, nitrite and phosphate, while no effects were observed on pH and dissolved oxygen. Analysis of 16S rRNA sequence data of water sample indicated significant (P < 0.05) shift of microbial abundance in post-fed FM and PBM water with the evidence of microbial transmission from the gut of marron. Post-fed marron resulted in a significant correlation of Hafnia, Enterobacter, Candidatus Bacilloplasma and Aquitella with the quality and microbial population of water. The results of this study generated valuable knowledge database of microbes-water relationship for better health management practices and production of marron aquaculture fed with FM and PBM diets in under restricted feeding regime with the feeding ratios provided.

Lactobacillus plantarum in black soldier fly (Hermetica illucens) meal modulates gut health and immunity of freshwater crayfish (Cherax cainii).

Probiotic supplements are being used to improve the growth and immune performance of aquaculture species over the last couple of decades. In recent times, black soldier fly (BSF) is considered as one of the promising sources of alternative protein to fishmeal protein in aqua-diets. Since the freshwater crayfish, marron (Cherax cainii), a Western Australian's native and iconic freshwater crayfish species, grows fairly slow under commercial farming environment, this study was aimed to investigate the supplemental effect of BSF and BSF with probiotic bacteria Lactobacillus plantarum (BSFLP) on overall health and immune performance of marron after 56 days of feeding under laboratory conditions. The post-trial data revealed insignificant influences of any diets on growth performance, however, both BSF and BSFLP based diets significantly improved some haemolymph parameters and gut health of marron. High throughput sequence data revealed that both BSF and BSFLP diets significantly improved the diversity of microbial communities including some beneficial bacteria for crustaceans in the hindgut of marron. Further analysis showed that both BSF and BSFLP diets upregulated the expression of some genes in the gut tissue and haemocytes associated with the innate immune response of marron at 48 h post injection. The up-regulation of some immune genes in BSFLP diet group was found significantly linked to OTU abundance for Lactobacillus. The findings of this study could be helpful for improving overall health status of marron.

The ameliorative effects of various fish protein hydrolysates in poultry by-product meal based diets on muscle quality, serum biochemistry and immunity in juvenile barramundi, Lates calcarifer.

In an effort to reduce the use of fishmeal (FM), the effect of using protein from poultry by product meal (PBM) along with the supplementation of three different fish protein hydrolysate (FPH) including yellowtail kingfish, carp and tuna hydrolysate (designated as KH, CH and TH, respectively) were evaluated in juvenile barramundi for growth performance, fillet quality, mucosal immunity, serum biochemistry, immune response and infection against Vibrio harveyi. Fish were fed a FM based control diet + three isonitrogenous and isolipidic diets containing 90% of PBM protein supplemented with different types of hydrolysates: 90% PBM +10% KH (90PBM + KH), 90% PBM + 10% CH (90PBM + CH) and 90% PBM + 10% TH (90PBM + TH). Growth performance and indices were unaffected by the hydrolysate supplemented diets when compared to the control. FPH supplemented PBM diets resulted in improved muscle quality by improving poly unsaturated fatty acids (PUFA), ∑n-3, ∑n-6 and ∑n-9, and health related lipid indexes were not affected. The internal architecture of spleen and kidney were not altered by test diets whilst FPH supplemented PBM modulated acidic mucins in intestine and skin of fish. Improved infection rate in response to two weeks post infection with V. harveyi in the FPH supplemented diets was further associated with an increased serum immune response and a concomitant regulation of proinflammatory and inflammatory cytokines in the head kidney. Serum biochemistry including alanine transaminase (ALT), glutamate dehydrogenase (GLDH) and total bilirubin (TB) showed a decreasing trend both in pre-challenge and post-challenge barramundi fed FPH supplemented diets whereas cholesterol level decreased significantly in post-challenge groups fed 90PBM + KH and 90PBM + TH than pre-challenge barramundi. This study signifies that supplementation of 10% with different three FPH, hydrolysed by an alcalase® enzyme in PBM-based diets for barramundi could be good strategies to overcome the negative consequences triggered by animal by-product ingredients.

Influence of fish protein hydrolysate produced from industrial residues on antioxidant activity, cytokine expression and gut microbial communities in juvenile barramundi Lates calcarifer.

The present study investigated the supplemental effects of tuna hydrolysate (TH) in poultry by-product meal (PBM) and dietary fishmeal (FM) diets on antioxidant enzymatic activities, gut microbial communities and expression of cytokine genes in the distal intestine of juvenile barramundi, Lates calcarifer. Fish were fed with fermented (FPBM + TH) as well as non-fermented PBM (PBM + TH) and FM (FMBD + TH) diets with 10% TH supplementation for 10 weeks. A basal diet prepared without TH supplementation served as control. The results showed that the activity of glutathione peroxidase was significantly higher in FPBM + TH than the control, while the malondialdehyde and catalase activities were unchanged. FPBM + TH diet significantly (P < 0.05) upregulated the pro-inflammatory cytokines including IL-1ß and TNF-α while considerable downregulation (P < 0.05) was observed in the mRNA expression levels of anti-inflammatory cytokine, IL-10 in the distal intestine of fish. The 16SrRNA analysis using V3-V4 region evidenced the ability of FPBM + TH to modulate the distal intestinal gut microbiome, augmenting the richness of Firmicutes and Fusobacteriaat at phylum level and Bacillus, Lactococcus and Cetobacterium at genus level. All these results have shown that fermented PBM with TH supplementation could improve the antioxidant capacity and inflammatory responses of juvenile barramundi while influencing the microbial communities at both phylum and genera levels.

Beneficial effects of tuna hydrolysate in poultry by-product meal diets on growth, immune response, intestinal health and disease resistance to Vibrio harveyi in juvenile barramundi, Lates calcarifer.

This study was conducted to investigate the effects that tuna hydrolysate (TH) supplementation in poultry by-product meal (PBM) diets would have on growth, immunity and resistance to Vibrio harveyi infection in juvenile barramundi, Lates calcarifer. Five isonitrogenous and isocaloric diets containing fishmeal (FM) without TH supplementation (control) and four diets with 10% TH supplementation viz. a FM protein diet (FMBD + TH), a 75% PBM protein diet (LPBM + TH) and two 90% PBM protein diets, either bioprocessed (BPBM + TH) or unprocessed (HPBM + TH), were formulated for juvenile barramundi, Lates calcarifer. The diets were fed to triplicate groups of juvenile barramundi (average pool weight 12.63 ±â€¯0.11 g) for 10 weeks. Significantly (P < 0.05) higher final body weights and specific growth rates were noted in fish fed with FMBD + TH and BPBM + TH diets when compared to the control. Transmission electron microscopy observation of fish distal intestines revealed a significant enhancement of microvilli length in fish fed FMBD + TH and BPBM + TH whereas scanning electron microscopy analysis found no significant difference in microvilli density. A bacterial challenge with Vibrio harveyi was conducted for 14 days after the growth trial to test the immune response and survival of barramundi. In the pre-challenge condition, a significant reduction in blood glucose was found in BPBM + TH compared to the control, and fish in the post-challenge at 24 h had higher glucose levels compared to fish in the pre- and post-challenge conditions at 72 h. The serum lysozyme activity was significantly higher in FMBD + TH and BPBM + TH compared to the control and fish at 72 h post-challenge exhibited higher lysozyme activity in each treatment compared to all dietary groups in the post-challenge condition at 24 h and to HPBM + TH and BPBM + TH in the pre-challenge condition. Fish fed FMBD + TH, LPBM + TH and BPBM + TH diets had significantly higher survival to the bacterial challenge than fish in the control and HPBM + TH. These results showed that PBM supplemented with TH could successfully replace FM without compromising growth, however, bioprocessed PBM supplemented with TH (BPBM + TH) may significantly improve growth performance, immune response, intestinal health and disease resistance in juvenile barramundi.

Effects of different dietary protein sources on the immunological and physiological responses of marron, Cherax cainii (Austin and Ryan, 2002) and its susceptibility to high temperature exposure.

A two phased feeding trial was conducted to evaluate the effects of alternative protein sources on the immunophysiological responses of marron. During the phase I, marron were fed with five alternative protein supplemented diets for 90 days, while in phase II, the same marron were exposed to elevated temperature (30 °C) and their immunophysiological responses were investigated post exposure. Five isoproteic (crude protein 30%) and isoenergetic diets were prepared by containing fishmeal, poultry by-product meal, feather meal, lupin meal, and meat and bone meal as the main protein source. A hundred and fifty juvenile marron (Cherax cainii) of the average weight 9.09 ±â€¯0.21 g were randomly distributed into 15 tanks (three replicates per feeding treatments). In the Phase I, general immune response parameters, such as, total haemocyte count (THC), proportion of hyaline cells, neutral red retention time (NRRT), phagocytic rate (PR), heamolymph bacteraemia, and condition indices of marron were investigated. The highest (P < 0.05) THC among dietary protein sources was obtained in marron fed with PbM at the end of experiment. Marron fed with FeM protein sources resulted in the highest survival rate followed by PbM fed group. Longer microvilli length (3.83 ±â€¯0.18 µm) was demonstrated in marron fed with PbM diet. Diets containing FM and PbM protein sources revealed significantly (P < 0.05) lower number of microvilli/group than diets containing FeM and LM. The results demonstrated that different dietary protein sources in the marron diets did not detect significant (P > 0.05) change of the condition indices throughout the experiment period, however highest Hiw and Hid was recorded in marron fed with PBM at day 45. The PR of marron fed dietary protein from PbM did not change significantly after temperature exposure. Increased NRRT, PR and haemolymph bacteraemia was observed with dietary feeding of FM at the end of the trial. However, results revealed that PbM could be an alternative protein source for culture of marron as reflected in terms of increased THC, longer microvillus length and improved susceptibility to high temperature exposure. Overall, result could serve as useful baseline data in developing cost effective potential diets for marron aquaculture.

Effects of the dietary supplementation of mixed probiotic spores of Bacillus amyloliquefaciens 54A, and Bacillus pumilus 47B on growth, innate immunity and stress responses of striped catfish (Pangasianodon hypophthalmus).

The study used the mixed probiotics of Bacillus amyloliquefaciens 54A and B. pumilus 47B isolated from striped catfish (Pangasianodon hypophthalmus) intestine aiming to stimulate growth performance, innate immunity, stress tolerance of striped catfish. The average weight gain (AWG), specific growth rate (SGR), and feed conversion ratio (FCR) were analyzed after fish were fed the mixture of probiotics (B. amyloliquefaciens 54A and B. pumilus 47B) at concentrations of 1 × 108, 3 × 108, and 5 × 108 CFU g-1 feed for 90 days. Immunity parameters, survival rate of fish challenged with Edwardsiella ictaluri and ammonia tolerance were also investigated. The amounts of B. amyloliquefaciens and B. pumilus were counted and identified by specific primer pairs of Ba1-F/Ba1-R, and 16-F/Bpu-R to confirm the presence of probiotics in fish intestine. The AWG (476.6 ± 7.81 g fish-1) of fish fed probiotics at 5 × 108 CFU g-1 was significant higher than the control (390 ± 25.7 g fish-1) after 90 days of feeding, but there was no significant (P > 0.05) effect of probiotics on FCR and SGR. Fish fed diet containing probiotics at 5 × 108 CFU g-1 also expressed resistance to E. ictaluri infection and higher immune parameters such as phagocytic activity, respiratory bursts, and lysozyme activity than the control. Stress response with ammonia showed significantly lower mortality rate (25%, 20% and 27%) of fish fed probiotics at all three levels of 1, 3 and 5 × 108 CFU g-1 than the fish fed control diet (75%). The study also demonstrated that the probiotics survived in the intestine of striped catfish after 90 days of feeding. Therefore, the dietary supplementation of a mixture of B. amyloliquefaciens and B. pumilus at 5 × 108 CFU g-1 can be used to improve the health and growth rate of striped catfish.

Growth, enzymatic glutathione peroxidase activity and biochemical status of juvenile barramundi (Lates calcarifer) fed dietary fermented soybean meal and organic selenium.

Solvent-extracted soybean meal (SBM) was fermented using baker's yeast Saccharomyces cerevisae at 30 °C for 5 days. Four isonitrogenous and isocaloric diets containing 75% SBM protein, either fermented or non-fermented (SBM and FSBM), and supplemented or not with organic Se (OS) (SBMOS and FSBMOS), were fed to triplicate groups of juvenile barramundi (Lates calcarifer) (initial weight of 5 g) for 75 days. A fishmeal (FM)-based diet formulated for juvenile barramundi was used as a reference diet. The growth of fish was significantly affected by either the interaction of SBM type or by the OS level. In fish fed diets supplemented with OS (SBMOS and FSBMOS), final weight (FW), specific growth rate (SGR) and weight gain (WG) were higher in fish fed the fermented SBM (FSBMOS) than in those fed the non-fermented SBM (SBMOS). The apparent digestibility coefficient (ADC) of protein was higher in the fish fed the fermented SBM, either supplemented or unsupplemented with OS. However, there were no significant differences in the ADC of dry matter (DM) and lipids among the tested diets and in comparison to the reference diet. The haematocrit and leucocrit of fish fed the FSBMOS diet were lower than those of fish fed the FM diet. Furthermore, glutathione peroxidase (GPx) activity was significantly influenced by OS supplementation in the experimental diets; GPx activity was greater in the fish fed diets supplemented with OS. Creatinine kinase (CK) of all groups of fish was higher than the CK of those fed the reference diet. These results suggest that with a proper nutritional level, OS supplementation may act as an important factor in enzymatic GPx activity and in the haematology and blood biochemistry status of juvenile barramundi fed fermented SBM-based diets, encouraging improvement of the overall growth performance.

Effects of carbon source addition in rearing water on sediment characteristics, growth and health of cultured marron (Cherax cainii).

Carbon sources are considered as critical input for the health and immunity of aquatic animals. The present study investigated the impact of different carbon sources on water quality parameters, carbon to nitrogen (C/N) ratio and microbial community in sediments, and health responses of marron (Cherax cainii) under laboratory conditions. Following one week of acclimation, 120 marron were randomly assigned to 12 experimental tanks. There were four treatments including one untreated control and three groups with carbon addition to maintain a C/N ratio of 12 maintained in culture water. Carbon supplementation groups included corn flour (CBC12), molasses (MBC12) and wheat flour (WBC12). At the end of the 60-day trial, MBC12 resulted in the highest sediment C/N ratio, followed by CBC12. Weight gain and specific growth rate were higher in MBC12, compared to control. The protease activity in marron hepatopancreas, total haemocyte count and lysozyme activity in haemolymph were highest in MBC12. Analysis of 16S rRNA sequence data of tank sediments revealed increased bacterial alpha diversity in MBC12 and WBC12. Proteobacteria was the most abundant phylum in MBC12 (88.6%), followed by control (82.4%) and CBC12 (72.8%). Sphingobium and Novosphingobium were the most abundant genera in control and MBC12 groups, respectively. Higher Aeromonas abundance in CBC12 and Flavobacterium in WBC12 were observed. Overall results indicated that MBC12 led to improved water quality, retaining high C/N ratio and enriched the bacterial populations in sediments resulting in improved growth and immune performance of marron.

Prevalence of opportunistic pathogens and anti-microbial resistance in urban aquaculture ponds.

Bacterial antimicrobial resistance (AMR) has emerged as a significant concern worldwide. The microbial community profile and potential AMR level in aquaculture ponds are often undervalued and attract less attention than other aquatic environments. We used amplicon and metagenomic shotgun sequencing to study microbial communities and AMR in six freshwater polyculture ponds in rural and urban areas of Bangladesh. Amplicon sequencing revealed different community structures between rural and urban ponds, with urban ponds having a higher bacterial diversity and opportunistic pathogens including Streptococcus, Staphylococcus, and Corynebacterium. Despite proteobacterial dominance, Firmicutes was the most interactive in the community network, especially in the urban ponds. Metagenomes showed that drug resistance was the most common type of AMR found, while metal resistance was only observed in urban ponds. AMR and metal resistance genes were found mainly in beta and gamma-proteobacteria in urban ponds, while AMR was found primarily in alpha-proteobacteria in rural ponds. We identified potential pathogens with a high profile of AMR and metal resistance in urban aquaculture ponds. As these ponds provide a significant source of protein for humans, our results raise significant concerns for the environmental sustainability of this food source and the dissemination of AMR into the food chain.

Immunological responses of customised probiotics-fed marron, Cherax tenuimanus, (Smith 1912) when challenged with Vibrio mimicus.

A two-phased experiment was conducted to investigate the effects of dietary supplementation of customised probiotics on marron physiology. During the first phase marron were fed probiotic supplemented feed for 70 days, while in phase two the same marron were challenged with Vibrio mimicus and their physiological responses were investigated for 4 days post-challenged. The experiment was carried out in a purpose-built room, designed for aquaculture research, using 18 of 250 L cylindrical plastic tanks. Five species of isolated probiotic bacteria from commercial probiotic products and marron's intestine were tested in this experiment. The probiotic bacteria were (Bacillus sp.); A10 (Bacillus mycoides); A12 (Shewanella sp.); PM3 (Bacillus subtilis); and PM4 (Bacillus sp.), which were added to the formulated basal marron diet (34% crude protein, 8% crude lipid, 6% ash) at a concentration of 10(8) cfu/g of feed. Immune responses of marron fed probiotics were evaluated by investigating organosomatic indices, growth rate, survival, intermoult period, total haemocytes counts (THC), proportion of granular cells (GC), bacteraemia, bacteria load in the intestine and water quality. The results showed that dietary supplementation of probiotics in marron had no significant impact on growth, intermoult period and survival of the marron. However, their supplementation improved the physiological condition of marron in terms of significantly higher tail muscle indices, THC and proportion of granular cells (GC) and reduced bacterial load in the haemolymph. The addition of probiotics in marron diets also increased the bacteria load in the marron intestine. In addition, dietary supplementation of the customised probiotics was effective in improving the resistance of marron against V. mimicus as they had higher THC, higher proportion of GC and lower presence of bacteria in their haemolymph, after marron were challenged with V. mimicus. The results also showed that probiotic Bacillus mycoides (A10) and PM4 are the most beneficial dietary probiotics for marron health.

Dietary organic selenium improves growth, survival and resistance to Vibrio mimicus in cultured marron, Cherax cainii (Austin, 2002).

To determine the effects of dietary organic selenium (OS) supplementation on the growth performance and immune competence of marron, Cherax cainii (Austin, 2002), a group of marron were fed 0.2 g kg(-1) of Sel-Plex(®) supplemented basal diet and then compared with another group (control) of marron fed basal diet without any supplementation. After 90 days of feeding, final weight, average weekly gains (AWG), relative gain rate (RGR), specific growth rate (SGR), survival, total and differential haemocyte counts (THC and DHC), were compared between the two groups. Surviving marron from each group were then divided into three sub-groups (three tanks per sub-group with seven marron per tank); (1) first sub-group was injected with 20 µL of 3.24 × 10(6) cfu Vibrio mimicus; (2) the second sub-group was injected with 20 µL normal saline and (3) the third sub-group was not subjected to injection and became the control group. THC, DHC, neutral red retention time (NRRT) and Vibrio ranks of post-injected marron were evaluated for 96 h, at every 24-h interval. The results showed that after 90 days of feeding, final weight, AWG, RGR, SGR, survival, THC, proportion of hyaline cells of OS-fed marron were significantly higher (P < 0.05) than the control group, whereas proportion of granular and semigranular cells were not affected by dietary OS. After challenging with V. mimicus, survival rate of marron without dietary OS significantly decreased (P < 0.05) as compared to the control group of marron. THC of marron in all sub-groups were significantly reduced (P < 0.05) after the challenge. However, THC and granular cells of sub-groups fed OS were higher than other sub-groups. Vibrio ranks and NRRT of marron fed OS were significantly lower and slower, respectively, than marron fed without OS. These findings demonstrated the benefits of OS inclusion in the marron diet in terms of growth, health and disease resistance.

A Mixture of Full-Fat and Defatted Hermetia illucens Larvae and Poultry By-Products as Sustainable Protein Sources Improved Fillet Quality Traits in Farmed Barramundi, Lates calcarifer.

The physicochemical quality and shelf-life of fillets from barramundi, which were fed for 56 days on a mixture of poultry by-product meal (PBM), full-fat Hermetia illucens (FHI), and defatted HI (DHI), were investigated and compared to a fishmeal (FM) control diet. The proximate and total amino acids compositions of the fillets were unaffected by the test diets, while the mixture of PBM and HI larvae improved the sensory quality. An eight-day shelf-life study showed that PBM-HI-based diets improved the texture profile based upon the chewiness, cohesiveness, gumminess, and hardness, regardless of the storage time. The improved texture was aligned with comparatively less degradation of the microstructure of the muscle tissue in the same diets. An improvement in the quality index (QI) value, an increase in pH, and a decrease in lipid oxidation were also found in the fillets of barramundi fed test diets compared with the control diet during the storage time. The test diets positively influenced flesh lightness and redness, while the color profiles were negatively influenced by the storage time. Overall, the maintenance of compositional attributes; the enhancement of fillet sensory attributes, texture, and brightness; and the improved raw fillet shelf-life support the inclusion of PBM-HI-based diets in aquafeed.

Fish waste to sustainable additives: Fish protein hydrolysates alleviate intestinal dysbiosis and muscle atrophy induced by poultry by-product meal in Lates calcarifer juvenile.

Valorising waste from the processing of fishery and aquaculture products into functional additives, and subsequent use in aquafeed as supplements could be a novel approach to promoting sustainability in the aquaculture industry. The present study supplemented 10% of various fish protein hydrolysates (FPHs), obtained from the hydrolysis of kingfish (KH), carp (CH) and tuna (TH) waste, with 90% of poultry by-product meal (PBM) protein to replace fishmeal (FM) completely from the barramundi diet. At the end of the trial, intestinal mucosal barriers damage, quantified by villus area (VA), lamina propria area (LPA), LPA ratio, villus length (VL), villus width (VW), and neutral mucin (NM) in barramundi fed a PBM-based diet was repaired when PBM was supplemented with various FPHs (p < 0.05, 0.01, and 0.001). PBM-TH diet further improved these barrier functions in the intestine of fish (p < 0.05 and 0.001). Similarly, FPHs supplementation suppressed PBM-induced intestinal inflammation by controlling the expression of inflammatory cytokines (tnf-α and il-10; p < 0.05 and 0.001) and a mucin-relevant production gene (i-mucin c; p < 0.001). The 16S rRNA data showed that a PBM-based diet resulted in dysbiosis of intestinal bacteria, supported by a lower abundance of microbial diversity (p < 0.001) aligned with a prevalence of Photobacterium. PBM-FPHs restored intestine homeostasis by enhancing microbial diversity compared to those fed a PBM diet (p < 0.001). PBM-TH improved the diversity (p < 0.001) further by elevating the Firmicutes phylum and the Ruminococcus, Faecalibacterium, and Bacteroides genera. Muscle atrophy, evaluated by fiber density, hyperplasia and hypertrophy and associated genes (igf-1, myf5, and myog), occurred in barramundi fed PBM diet but was repaired after supplementation of FPHs with the PBM (p < 0.05, 0.01, and 0.001). Similarly, creatine kinase, calcium, phosphorous, and haptoglobin were impacted by PBM-based diet (p < 0.05) but were restored in barramundi fed FPHs supplemented diets (p < 0.05 and 0.01). Hence, using circular economy principles, functional FPHs could be recovered from the fish waste applied in aquafeed formulations and could prevent PBM-induced intestinal dysbiosis and muscular atrophy. GRAPHICAL ABSTRACT.

A Mixture of Manganese, Silica and Phosphorus Supplementation Alters the Plankton Density, Species Diversity, Gut Microbiota and Improved the Health Status of Cultured Marron (Cherax cainii, Austin and Ryan, 2002).

Trace element supplementation to the freshwater environment can influence the plankton density and species diversity, contributing to the nutrition of aquaculture species, especially during the juvenile stage. An experiment was conducted under laboratory conditions to evaluate the effects of supplementing different mixtures of manganese, silica and phosphorus on the plankton density and species diversity and their impact on cultured juvenile marron (Cherax cainii, Austin and Ryan, 2002). Manganese, silica and phosphorus in concentrations of 0.0024, 0.41, 0.05 mg*L-1; 0.0041, 0.82, 0.12 mg*L-1; and 0.0058, 1.26, 0.25 mg*L-1 respectively termed as low, medium and high were supplemented to tank water containing a phytoplankton density of 3.77 ± 0.16 × 106 cells*L-1 and 292.9 ± 17.6 individuals*L-1 of zooplankton, and plankton growth was observed every 24 h for 6 days. Afterwards, a 3-month trial was conducted studying the effects of these trace element concentrations and resulting plankton densities on marron growth, survival, moulting, gut microbiota and health indices. Silica supplementation at high concentration increased the diatom abundance, silica and phosphorus supplementation at higher concentration that resulted in a significant increase in plankton density and species diversity, leading to improved marron health indices than the control and the tanks receiving a low concentration. Marron-specific growth rate, weight gain and dissolved copper concentration in haemolymph were significantly higher in tanks with higher supplementation and higher plankton density. Marron survival, moult interval and total haemocyte count were not affected by the supplementation. Marron gut microbiota at higher trace element concentration supplementation showed a significant increase in abundance of phosphate solubilizing bacteria.

Sources of protein diet differentially stimulate the gut and water microbiota under freshwater crayfish, marron (Cherax cainii, Austin 2002) culture.

To reduce the reliance on fishmeal (FM), other protein sources have been evaluated on cultured animals. In a 60-days feeding trial, marrons (Cherax cainii) were fed a FM diet and five test diets containing 100% of plant-based protein sources such as soybean, lupin and valorised animal-based proteins such as poultry-by-product, black soldier fly and tuna hydrolysate. At the end of the trial, DNA samples from marron gut and rearing water were investigated through DNA-based 16S rRNA gene sequencing. Plant-based diets increased abundance for Aeromonas, Flavobacterium and Vogesella, whereas animal and insect proteins influenced diverse bacterial groups in the gut linked to various metabolic activities. Insect meal in the water favoured the growth of Firmicutes and lactic acid bacteria, beneficial for the marron health. Aeromonas richness in the gut and reared water signified the ubiquitous nature of the genus in the environment. The higher bacterial diversity in the gut and water with PBP and BSF was further supported by qPCR quantification of the bacterial single-copy gene, rpoB. The overall results suggested that PBP and BSF can exhibit positive and influential effects on the gut and water microbial communities, hence can be used as sustainable ingredients for the crayfish aquaculture.

Zeolite mediated processing of nitrogenous waste in the rearing environment influences gut and sediment microbial community in freshwater crayfish (Cherax cainii) culture.

Zeolite is known to uptake toxic metals and filter nitrogenous waste from aquaculture effluents. The present study aimed to investigate the impacts of zeolite in three different applications namely, dietary zeolite (DZ), suspended zeolite (SZ) in the water column, and a combination of both (DZSZ) relative to unexposed freshwater crayfish, marron (control). At the end of the 56-days trial, the impact was assessed in terms of characterization of microbial communities in the culture environment and the intestine of marron. Alongside the microbial communities, the innate immune response of marron was also evaluated. The 16S rRNA data showed that marrons exposed to the suspended zeolite had a significant increase of bacterial diversity in the gut, including the restoration of marron core operational taxonomic units (OTUs), relative to other forms of exposures (DZ, DZSZ) and the control. Suspended zeolite alone also increased the number of unshared OTUs and genera, and improved predicted metabolic functions for the biosynthesis and digestion of proteins, amino acids, fatty acids, and hormones. In the tank sediment, the shift of microbial communities was connected more strongly with the time of experiment than the type of zeolite exposure. In the second case, only control marron had a different microbial ordination in terms of rare taxa present in the community. Nevertheless, the modulation in the gut environment was found more prominent in DZ, relative to modulation in the tank sediments. The taxa-environment correlation identified Rhodoferax as the most potential bacteria in removing nitrogenous waste from the rearing environment. Further analysis showed that SZ resulted in the upregulation of genes associated with the innate immune response of marron. Overall results suggest that SZ can be used to enrich microbial communities in the gut and tank sediments and better immune performance of marron.