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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.

Two experiments were conducted to evaluate the efficacy of an oil-based substrate, The Water Cleanser™ (TWC). The first experiment studied the effects two substrates of different oil composition (TWC, TWC +), and a commercial bacterial additive (Bio-Aid) on concentrations of nitrogen and phosphorous in indoor aquaria. The second experiment studied the effects of TWC, a bacterial additive, and a combination (TWC + B) on concentrations of nitrogen and phosphorous, phytoplankton abundance and diversity in outdoor freshwater crayfish (Cherax cainii, Austin and Ryan, Invertebr Syst 16:357–367, 2002) tanks. In the first experiment, the concentration of TAN decreased more rapidly with Bio-Aid, whilst the maximum concentrations of NO2-N and NO3-N were reduced with the substrates. The concentration of orthophosphate was reduced in aquaria with TWC + . In the second experiment, concentrations of TAN, NO2-N, NO3-N and total phosphate were not significantly affected by TWC. After addition of TWC + B, there was a significant decrease in the concentrations of nitrate and total phosphate, and a higher abundance of phytoplankton was maintained than with other treatments. Additionally, a population Bacillus sp. was found on the substrate surface. TWC had no adverse effects on phytoplankton abundance or diversity, or C. cainii physiology, weight gain or survival. TWC and TWC + were effective bioremediators of eutrophic water, whilst a combination of TWC + B was effective in short term bioremediation and in promoting phytoplankton abundance in C. cainii tank culture.

An evaluation of six dietary protein to energy (P/E) ratios was conducted in the early juveniles of ornate spiny lobster (Panulirus ornatus) by investigating the effects on physiology, digestive enzyme activities, body composition and amino acid profiles. Lobster juveniles (0.25 ± 0.01 g) were individually kept in aquaria and fed 6 isoenergetic formulated diets consisting of three protein levels (450, 500 and 550 g kg− 1) and three lipid levels (7, 9 and 13 g kg− 1) having P/E ratios of 21, 22, 23, 24, 25 and 26 mg MJ− 1. After 8 weeks of feeding, the dietary P/E ratio influenced the specific growth rates, while survival, moult increment, moulting rate and moult interval were not affected by any dietary P/E ratios. The dietary protein levels had stronger influence on the specific growth rate whereas dietary lipid levels affected the length increments. The dietary P/E ratio also influenced the protease, lipase, super oxidase dismutase activities and the body composition of spiny lobster juveniles. Both protein and lipid in the whole body of the juveniles reflected the dietary composition. The total quantity of non-essential amino acids was higher than essential amino acids in the formulated diet and also in the body of juvenile lobsters. Results clearly indicated that 55% and 9% dietary protein and lipids, respectively with 26.6 g CP MJ− 1 can yield the maximum growth and digestive enzyme activities.

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.

The abundance and diversity of the plankton community represents the health of the aquatic ecosystem, and plays an important role in the growth of cultured animals under aquaculture conditions. The temporal variations of plankton abundance, taxonomic composition, diversity, evenness and species richness were studied in three old and three new semi-intensive marron (Cherax cainii, Austin and Ryan, 2002) ponds. Water parameters such as temperature, dissolved oxygen, pH, turbidity, TAN, nitrite, nitrate and reactive phosphate were recorded, and plankton samples were collected every two months, for one year of juvenile production cycle. A total of twenty-six phytoplankton and seven zooplankton genera were recorded. Chlorophyceae was the dominant class of phytoplankton throughout the year, followed by Trebouxiophyceae. Rotifera comprised 49.8% of the total zooplankton community (individuals L−1), the largest proportion of any group. Temporal variations impacted the plankton abundance and community structure, and plankton abundance were more abundant during summer. The pond age did not influence the phytoplankton abundance, whereas zooplankton abundance was higher in older ponds.

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

The search for suitable fish meal replacements in aqua-diets is a salient agenda in the constant effort of making aquaculture practices more sustainable. In this study, we tested four customised diets composed by systematic inclusion of pre-selected fish meal substitutes, lupin kernel meal, BSF meal, TH and PBM on growth, metabolism, cytokine profile, gut morphology and microbiota of juvenile Lates calcarifer. Five isoproteic and isoenergetic diets were prepared viz. FM100 as a control (without fish meal substitute), while FM75, FM50, FM25 and FM0 indicates replacement of fish meal (FM) at 25%, 50%, 75%, and 100%, respectively by a mixture of four different pre-selected non-fish meal (NFM) ingredients. Fish fed FM100, FM75, FM50, FM25 exhibited consistent growth and haematological response, while the fish fed no fishmeal (FM0) showed significant decline in final body weight (FBW) and specific growth rate (SGR). The poor growth performance was correlated with a decrease in villous width, microvilli height and goblet cells density. A significant shift in abundance profile of Psychrobacter in the gut microbial profile of fish fed FM50 was noticed compared to fish fed FM100. The results of qRT-PCR showed up-regulated expression of innate immune responsive genes in the FM50 group. The adverse impacts on growth performance and gut health of fish fed FM0 suggest that the complete substitution of fishmeal is not advisable and the inclusion range of these alternatives should be decided for a species only after examining their effect on maximal physiological performance.

In intensive farming systems, fish are held at high densities, which may increase stress, leading to susceptibility to diseases that result in economic losses. Therefore, effective feeding practices incorporating health‐promoting compounds such as proteins, hydrolysates and bioactive peptides that can stimulate the defence mechanisms of fish and achieve better growth are some of the priorities for sustainable aquaculture development. Globally, the fish processing industries generate and discard a large volume of waste every year, estimated at up to 60% of the harvested biomass. This waste can be converted to value‐added products such as fish protein hydrolysate (FPH) with the addition of various proteolytic enzymes. FPH from fish processing waste including skin, heads, muscle, viscera, liver and bones is a good source of protein, amino acids, peptides and antioxidants and has been found to possess desirable functional and bioactive peptides. A moderate inclusion of FPH in aquafeeds has the potential to improve growth, feed utilization, immune functions and disease resistance of fish. Production of FPH, targeted to more precise molecular weight ranges, has superior functionalities that are in high demand. With interest in FPH as an aquafeed supplement, this review aimed to summarize the source, production processes and functional properties of FPH and the reported impact of FPH in aquafeed supplement on fish growth, survival, feed utilization, immune response and disease resistance. Possible limitations of using FPH and future research potential as an opportunity for the use of processing fish waste are also discussed.

Seasonal variations can affect the concentration of trace elements, and the change in their concentrations can affect the natural productivity of freshwater aquaculture ponds. Hence, we studied the seasonal variations of the 12 pre‐selected trace elements (Co, Cu, Fe, Mn, Zn, Se, Ca, Mg, P, S, Al, Si) and their relationships with primary and secondary productivity in two aged ponds, stocked with three different life stages of marron (Cherax cainii), for a period of 1 year. Trace element analysis was performed by using (Agilent, ICP‐OES). Except Co and Se, all trace elements, and primary and secondary productivity were influenced by seasonal variation. The pond age significantly influenced the concentrations of some trace elements. On a seasonal basis, trace elements were positively correlated with the plankton abundance, species diversity, and wet and dry plankton weights. Seasonal variations and pond age affected the dissolved concentrations of trace elements and plankton productivity.

This study aimed to investigate the combined effects of two most potent probiotic bacteria Lactobacillus acidophilus and Lactobacillus plantarum on overall health and immune status of freshwater crayfish, marron under laboratory conditions. A total of 36 marron were distributed into six different tanks and two different feeding groups, control and probiotic-fed group. After acclimation, control group was fed with basal diet while probiotic group was fed 109 CFU/mL per kg of bacterial supplemented feed for 60 days. The results showed no significant differences in weight gain, however, probiotic feed significantly enhanced some hemolymph parameters and biochemical composition of tail muscle. Histology data revealed better hepatopancreas health and higher microvilli counts in the marron gut fed probiotic diet. The probiotic bacteria triggered significant shift of microbial communities at different taxa level, mostly those reported as beneficial for crayfish. The probiotic diet also enriched the metabolic functions and genes associated with innate immune response of crayfish. Further correlation analysis revealed significant association of some taxa with increased activity for hemolymph and immune genes. Therefore, dietary Lactobacillus supplementation can modulate the overall health and immunity as well as gut microbial composition and interaction network between gut microbiota and immune system in crayfish.

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-1β 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.

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.

Black soldier fly (Hermentia illucens) meal has become a promising alternative substitution of fishmeal as protein resources in aquafeeds. However, the effect of defatted BSF meal inclusion on the juvenile lobster feed stability have not been assessed. The present study is aimed to evaluate the effects of the inclusion of defatted BSF meal (25, 35 and 50 % replacement), the salinity level of seawater (30 and 35 ppt) and the immersion time (15, 30, 60, 90 and 120 minutes) on the pellet stability of formulated feed for juvenile lobster. The pellet stability was determined by calculate the dry matter loss of the feed (wet durability test). Results has shown that the dry matter loss of the feed ranged from 3.17 ± 0.03 % to 4.87 ± 0.26 % for the salinity of 30 ppt, and 3.27 ± 0.04 % to 4.46 ± 0.66 % for the salinity of 35 ppt. The dry matter loss was affected by the levels of defatted BSF inclusion level, the salinity, and the immersion time. The means of dry matter loss in formulated feeds were conversely affected by the salinity level and consistent in all formulated feeds. The means of dry matter loss in BSF25, BSF35 and BSF50 feed were significantly affected by the immersion time (P

Aquaculture has rapidly developed and expanded, leading to high demand on fishmeal, which is the main protein source for aqua-feed production. Some alternative protein sources have been evaluated to replace fishmeal in aquaculture. Fish protein hydrolysates (FPH) are prepared by enzymatic hydrolyzing fish protein in fish wastes discarded from the fish processing industry. FPH are promising alternative protein sources because they not only contain high protein level, balanced amino acid profile, and antioxidants; but also possess desirable functional and bioactive peptides. The review summarized the potential effects of fish protein hydrolysate (FPH) as supplementation on the growth performance, feed utilization, and physiological and immunological response of fish species. Moderate dietary FPH levels could improve growth, feed utilization and disease resistance in fish. However, excessive FPH inclusion levels led to the depression of growth and health of fish. The data suggested that supplementation of optimum FPH levels in diets may take their full effective utilization in the enhancement of growth and health of fish.

The proximate composition, sensory attributes, and shelf life of filets from barramundi, Lates calcarifer, were fed a fishmeal (FM) based diet (0PBM-0HI) and three test diets replacing FM protein entirely with 85% poultry by-products meal (PBM) and 15% Hermetia illucens (HI) larvae meal protein (85PBM-15HI), 80% PBM and 20% HI (80PBM-20HI) and 75% PBM and 25% HI (75PBM-25HI) were investigated. After a 56-day feeding trial, the crude protein, moisture, and ash percentage were unchanged while the crude lipid increased in barramundi filet when fed with PBM-HI-based diets. The increase in C12:0 (lauric acid) and C14:0 (myristic acid) resulted in an increase in the total saturated fatty acid while the monounsaturated fatty acid elevated due to an increase in C16:1n7 and C18:1cis + trans in the filet of the barramundi fed with a PBM-HI based diet. While the decrease in the total polyunsaturated fatty acid (PUFA) content in PBM-HI based fed barramundi filet was mainly due to a decrease in essential fatty acids including C20:5n3 [eicosapentaenoic acid (EPA)] and C22:6n3 [docosahexaenoic acid (DHA)] when compared with the 0PBM-0HI fed barramundi filet. The sensory quality was improved by PBM-HI-based diets, manifested by the highest scores given by the panelists. Texture profiles were not affected by diet but cohesiveness, gumminess, and chewiness decreased with increasing storage time. On days 1 and 8, skin brightness decreased in the skin of the barramundi fed with 85PBM-15HI and 80PBM-20HI compared with the skin of the 0PBM-0HI fed barramundi. Skin redness improved in fish-fed PBM-HI-based diets. The flesh brightness and yellowness increased significantly in barramundi when fed with PBM-HI-based diets. On days 1 and 4, the flesh brightness of the barramundi fed with PBM-HI-based diets demonstrated an increase compared with 0PBM-0HI. PBM-HI diets suppress lipid oxidation while lipid oxidation increased over the storage time. In summary, the improvement in sensory quality and color coupled with the suppression of rancidity in barramundi filets underpinned the potentiality of using the mixture of PBM and HI transformed from food waste in the barramundi diet to improve the filet quality and thus support sustainability and circular economy in aquaculture., Graphical Abstract

This study investigated the effects of replacement of fishmeal (FM) with poultry by-product (PBM) protein, supplemented with black soldier fly, Hermetia illucens (HI) larvae on growth, histomormhology, immunity and resistance to Vibrio harveyi in juvenile barramundi. Two hundred and twenty five barramundi averaging 3.51 ± 0.03 g were randomly allocated into three groups and fed isonitrogenous and isocalorific diets containing different levels of PBM supplemented with HI as follows: Control (FM based diet), 45PBM + HI (45% PBM supplemented with 10% HI), and 90PBM + HI (90% PBM supplemented with 10% HI) for 6 weeks. Results showed that dietary inclusion of 45PBM + HI significantly improved the growth performance than control whereas growth inhibition occurred in the 90PBM + HI. The 45PBM + HI groups demonstrated significant increases in histometric measurements (villus and enterocyte width, and microvilli height) and acidic mucins. The impaired growth in 90PBM + HI groups was further associated with multifocal necrosis in the liver, an upregulation of the stress related genes (HSP70 and HSP90) and increase in the levels of liver enzymes. When 45PBM + HI was fed, survival against V. harveyi increased significantly and also an increase in serum immunity and immune-related genes in the head kidney was observed after infection.

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 ± 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.

AIMS: This study conducted bacterial community, virulence and antibiogram profiling inside the hindgut and skin of freshly caught hilsa fish and those sold at markets. METHODS AND RESULTS: The results of 16S rRNA‐based high‐throughput sequencing showed a higher number of bacterial genera in marketed fish samples than in fresh fish samples. The total operational taxonomic units, genus counts and diversity index were significantly higher (P > 0·05) in marketed fish, which also had abundant pathogenic bacterial groups. Skin samples had a lower profusion of pathogenic bacteria than gut samples. A total of 52 bacterial isolates from nine species were identified in this study, of which 25 were from a Chittagong market and 22 were from a Dhaka market, whereas only five were from fresh hilsa. The polymerase chain reaction amplification of 12 species‐specific virulence genes in the 52 isolates, namely, aer, hly, chxA, toxB, rtxC, sfa, uge, norB, trx, toxA, ipaH, sigA and coa, indicated a high number of positive samples containing Vibrio cholerae, Aeromonas spp., Klebsiella pneumoniae, Escherichia coli and Staphylococcus aureus. Antibiogram profiling of these bacteria against 10 commercial antibiotics showed high‐resistance patterns of the isolates against sulfamethoxazole, kanamycin, neomycin, ampicillin and tetracycline. CONCLUSION: The results reveal the spread of multidrug‐resistant bacteria in hilsa fish marketed for human consumption in Bangladesh. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the risk of spreading environmentally and clinically pathogenic bacteria in fish sold for human consumption in Bangladesh. Such bacteria come from aquatic pollution and poor handling, storage and transportation practices that may predispose fish to major outbreaks of infectious and waterborne diseases.

The effects of feeding different levels of poultry by-product meal (PBM) replacing fishmeal (FM) protein, supplemented with tuna hydrolysate (TH) and Hermetia illucens (HI) larvae, on the growth, fillet quality, histological traits, immune status, oxidative biomarker levels and gut microbiota of juvenile barramundi, Lates calcarifer were investigated for six weeks. Barramundi were fed four isonitrogenous and isolipidic diets in which a FM based diet was used as the Control diet (Diet1) and compared with other non-FM diets containing 80%, 85% and 90% PBM along with the concurrent supplementation of 5% and/or 10% TH and HI larvae meal. These treatment diets were designated as 80PBM10TH+10HI (Diet2), 85PBM5TH+10HI (Diet3) and 90PBM5TH+5HI (Diet4). The growth and condition factor of fish fed 80PBM10TH+10HI and 85PBM5TH+10HI were significantly higher than the Control. Total saturated, monounsaturated and polyunsaturated fatty acid retention in the fish muscle increased in fish fed PBM-based diets, supplemented with TH and HI larvae meal, with no adverse effect on post-harvest characteristics such as texture and colour of fish fillets. Improvement in serum total bilirubin and total protein content was found in all fish fed TH and HI larvae supplemented PBM. Similarly, immune response showed a significant increase in fish fed non-FM test diets than the Control. In the distal intestine, supplementation of any quantities of TH and HI larvae to PBM led to an increase in the microvilli density and neutral mucins while the number of goblet cells in the skin were unchanged. Liver, kidney, and spleen histology demonstrated a normal structure with no obvious changes in response to all test diets. Bacterial diversity increased in fish fed Diets 2 and 3 with a high abundance of Proteobacteria in Diets 1 and 4 and Firmicutes in Diets 2 and 3. The fish on test diets showed a lower abundance of genus Vibrio. Fish fed TH and HI larvae supplemented PBM diets showed lower infection rate to V. harveyi than the Control. Collectively, concurrent supplementation of TH and HI larvae could improve the quality of PBM diets with positive effects on growth, fillet quality, intestinal health, immunity, and disease resistance.

Fish farming including freshwater crayfish still has a dependency on the availability of fish meal as the main source of protein in feed. The purpose of this study was to evaluate alternative protein sources including poultry by-products, feather meal, lupine flour, soybean meal and meat, and bone meal for smooth marron (Cherax cainii) freshwater crayfish feed. A total of 150 juvenile marrons (9.09 ± 0.21 g) were kept for 90 days and distributed into 15 rearing tanks consisting of five treatments with three replications. The results indicated that there was no significant difference in the increase in biomass, survival rate, moulting rate (weight), feed efficiency ratio, and feed digestibility (P> 0.05). However, marron juvenile fed with a protein source of poultry by-products meal had the highest specific growth rate (0.31 ± 0.05 g / day). In addition, marron fed with fish meal protein resulted a significant increase in carapace length (P 0.05). Overall, the results of this study indicate that the general growth performance of marron is not affected by the replacement of fish meal in the feed. Alternative protein materials can be used as a protein source to replace fish meal in marron feed so that the use of fish meal can be reduced.

A 6-week feeding trial was performed to examine the effects of supplementing Hermetia illucens (HI) larvae meal when fishmeal (FM) was replaced with poultry by-product meal (PBM) in juvenile barramundi, Lates calcarifer diet. The effect was evaluated in terms of barramundi growth, filet quality, internal tissue structure, serum biochemistry, skin neutral mucins, immune response, and resistance to Vibrio harveyi. Three isonitrogenous (48% crude protein) and isolipidic (18% crude lipid) diets: an FM-based diet (control) and two diets containing 60 and 75% of PBM supplemented with 10% HI larvae (60PBM + HI and 75PBM + HI) were formulated. A total of 225 barramundi, with an average weight of 15.87 ± 0.14 g, were randomly distributed into nine tanks, each holding 25 fish. There were no significant effects of test diets on growth, but feeding HI-supplemented PBM diets significantly increased the survival rate. A significantly reduced intraperitoneal fat index in HI-supplemented-PBM-fed fish was correlated to a decreased size of peritoneal adipocytes. The observation of no histopathological alteration of the liver in the HI-supplemented-PBM-fed fish was further supported by significant alterations in serum biochemistry, in particular, a decreasing tendency of alanine transaminase, glutamate dehydrogenase, and total bilirubin. A 14-day challenge with V. harveyi indicated that HI-supplemented PBM diets reduced the infection rate in barramundi. After 24 h of infection, increased serum (lysozyme) and skin barrier functions, down-regulation of interleukin-1beta, and upregulation of interleukin-10 were found in HI-supplemented-PBM-fed fish.

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.

The abundance and diversity of the plankton community represents the health of the aquatic ecosystem, and plays an important role in the growth of cultured animals under aquaculture conditions. The temporal variations of plankton abundance, taxonomic composition, diversity, evenness and species richness were studied in three old and three new semi-intensive marron (Cherax cainii, Austin and Ryan, 2002) ponds. Water parameters such as temperature, dissolved oxygen, pH, turbidity, TAN, nitrite, nitrate and reactive phosphate were recorded, and plankton samples were collected every two months, for one year of juvenile production cycle. A total of twenty-six phytoplankton and seven zooplankton genera were recorded. Chlorophyceae was the dominant class of phytoplankton throughout the year, followed by Trebouxiophyceae. Rotifera comprised 49.8% of the total zooplankton community (individuals L−1), the largest proportion of any group. Temporal variations impacted the plankton abundance and community structure, and plankton abundance were more abundant during summer. The pond age did not influence the phytoplankton abundance, whereas zooplankton abundance was higher in older ponds.

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.

A feeding trial was carried out to evaluate the effects of substitution of fishmeal (FM) by dietary poultry by-product meal, fermented by Lactobacillus casei and Saccharomyces cerevisiae on growth, intestinal health, microbial composition, immune related cytokines and disease resistance of freshwater crayfish, marron (Cherax cainii) against Vibrio mimicus. Two isonitrogenous and isocaloric diets were formulated by replacing FM protein with fermented poultry by-product meal (FPBM) protein at 0% (Control) and 75% (FPBM), and fed marron for 70 days. The results indicated no significant difference (P > 0.05) in final body weights between two groups of marron, whilst intestinal microvilli number per fold was increased in marron fed FPBM than the control. The 16S rRNA sequences revealed an increased number of Lactobacillus and Streptococcus, and decreased number of Aeromonas at genus level in the distal intestine of marron fed FPBM. Marron fed FPBM showed up-regulated expression of IL-8, IL-10, and IL-17F genes in the distal intestine. Significantly (P < 0.05) increased lysozyme and phagocytic activity, and higher survival was found in marron fed FPBM following a bacterial challenge with Vibrio mimicus. Therefore, it is concluded that FPBM is beneficial to marron in terms of microbial community, immune-related cytokines and disease resistance against V. mimicus.