Characterization of hatchery residues for on farm implementation of circular waste management practices.

The conventional management of hatchery residues is associated with greenhouse gas and unpleasant odor emissions, the presence of pathogens and high disposal costs for producers. To address these issues, on-farm alternatives like composting, fermentation, and insect valorization are promising approaches. This study aims to characterize hatchery residues and define critical quality thresholds to identify effective processes for their management. Hatchery residue samples were collected bi-monthly over a year (N = 24) and were analyzed for proximate composition (dry matter, ash, energy, crude protein, crude lipid, crude fiber, carbohydrates), pH, color (L*a*b*, Chroma) and microbiological loads (total aerobic mesophilic counts, coliforms, lactic acid bacteria). Volatile fatty acid composition was also measured (N = 8). Significant correlation coefficients were found between TAM and LAB loads and residue characterization (pH, chroma, crude fibers, carbohydrates, and temperature). On a dry matter basis, residues were high in energy (2498 to 5911 cal/g), proteins (21.3 to 49.4 %) and lipids (14.6 to 29.1 %), but low in carbohydrates (0 to 15.3 %) despite temporal fluctuations. Ash content varied widely (8.6 to 49.1 %, dry matter) and is influenced by eggshell content. Microbiological loads were high for total aerobic mesophilic bacteria (6.5 to 9.1 log cfu/g), coliforms (5.4 to 8.5 log cfu/g) and lactic acid bacteria (6.7 to 9.0 log cfu/g). Valorization of hatchery residues on the farm will depends on the optimization of effective upstream stabilization processes. The critical points are discussed according to the valorization potentials that could be implemented on the farm from composting to upcycling by insects.

Apparent Digestibility of Macronutrients and Fatty Acids from Microalgae (Schizochytrium sp.) Fed to Rainbow Trout (Oncorhynchus mykiss): A Potential Candidate for Fish Oil Substitution.

Aquaculture feed formulation has recently turned its focus to reduce the reliance on marine-derived resources and utilise alternative feedstuffs, as an approach to improve the environmental sustainability of the aquaculture sector. The fish oil market is highly volatile, and availability of this commodity is continuously decreasing for use in aquaculture. Currently, a growing number of commercial efforts producing microalgae are providing omega 3-rich oil for sustainable aquaculture feed. This study was focused to determine the nutrient digestibility of a marine microalga, Schizochytrium spp., which is rich in docosahexaenoic acid (DHA) and long-chain polyunsaturated fatty acids (LC-PUFA), as a novel dietary lipid source that could be utilized effectively by rainbow trout (Oncorhynchus mykiss). A whole-cell Schizochytrium spp. biomass was used in the digestibility experiment at two different temperatures, 8 °C and 15 °C. No significant differences were detected between the two temperatures for the apparent digestibility coefficients (ADCs) of the dry matter (94.3 ± 4.9%), total lipids (85.8 ± 0.0%), crude proteins (89.5 ± 1.8%), energy (83.1 ± 1.7%) and fatty acids (85.8 ± 7.5%). The ADCs of the nutrients, energy, DHA and other fatty acids showed that Schizochytrium spp. is a high-quality candidate for fish oil substitution and supplement of LC-PUFA in fish feed with vegetable oils.

Thermal tolerance and fish heart integrity: fatty acids profiles as predictors of species resilience.

The cardiovascular system is a major limiting system in thermal adaptation, but the exact physiological mechanisms underlying responses to thermal stress are still not completely understood. Recent studies have uncovered the possible role of reactive oxygen species production rates of heart mitochondria in determining species' upper thermal limits. The present study examines the relationship between individual response to a thermal challenge test (CTmax), susceptibility to peroxidation of membrane lipids, heart fatty acid profiles and cardiac antioxidant enzyme activities in two salmonid species from different thermal habitats (Salvelinus alpinus, Salvelinus fontinalis) and their hybrids. The susceptibility to peroxidation of membranes in the heart was negatively correlated with individual thermal tolerance. The same relationship was found for arachidonic and eicosapentaenoic acid. Total H2O2 buffering activity of the heart muscle was higher for the group with high thermal resistance. These findings underline a potential general causative relationship between sensitivity to oxidative stress, specific fatty acids, antioxidant activity in the cardiac muscle and thermal tolerance in fish and likely other ectotherms. Heart fatty acid profile could be indicative of species resilience to global change, and more importantly the plasticity of this trait could predict the adaptability of fish species or populations to changes in environmental temperature.

Effects of Killing Methods on Lipid Oxidation, Colour and Microbial Load of Black Soldier Fly (Hermetia illucens) Larvae.

Black soldier fly (BSF) larvae represent a promising alternative ingredient for animal feed. Post-production processing can, however, affect their quality. This project aimed to optimize larval killing by comparing the effects on the nutritional and microbiological quality of 10 methods, i.e., blanching (B = 40 s), desiccation (D = 60 °C, 30 min), freezing (F20 = -20 °C, 1 h; F40 = -40 °C, 1 h; N = liquid nitrogen, 40 s), high hydrostatic pressure (HHP = 3 min, 600 MPa), grinding (G = 2 min) and asphyxiation (CO2 = 120 h; N2 = 144 h; vacuum conditioning, V = 120 h). Some methods affected the pH (B, asphyxiation), total moisture (B, asphyxiation and D) and ash contents (B, p < 0.001). The lipid content (asphyxiation) and their oxidation levels (B, asphyxiation and D) were also affected (p < 0.001). Killing methods altered the larvae colour during freeze-drying and in the final product. Blanching appears to be the most appropriate strategy since it minimizes lipid oxidation (primary = 4.6 ± 0.7 mg cumen hydroperoxide (CHP) equivalents/kg; secondary = 1.0 ± 0.1 mg malondialdehyde/kg), reduces microbial contamination and initiates dehydration (water content = 78.1 ± 1.0%). We propose herein, an optimized protocol to kill BSF that meet the Canadian regulatory requirements of the insect production and processing industry.

Hybridization between char species (Salvelinus alpinus and Salvelinus fontinalis): a fast track for novel allometric trajectories.

Hybridization between closely related species can generate genetic and phenotypic variation, providing valuable biological material to assess the physiological impact of the structural or functional variability of different organs. In the present study, we examined growth rates of various organs and whole body in brook char, Arctic char and their reciprocal hybrids over a period of 281 days. Parental species achieved significantly higher body mass than their hybrids. Hybridization significantly reduced the relative size of the heart, liver and spleen. The relative size of pyloric caeca did not differ among the four groups. The observed lower growth performance of the hybrids compared to parental species strongly suggests that divergence in the relative size of digestive organs, liver and heart partly dictate growth capacity. Our results also suggest that the increased variability achieved through hybridization may prove useful in a genetic selection program.

Thermal tolerance and thermal sensitivity of heart mitochondria: Mitochondrial integrity and ROS production.

Cardiac mitochondrial metabolism provides 90% of the ATP necessary for the contractile exertion of the heart muscle. Mitochondria are therefore assumed to play a pivotal role in heart failure (HF), cardiovascular disease and ageing. Heat stress increases energy metabolism and oxygen demand in tissues throughout the body and imposes a major challenge on the heart, which is suspected of being the first organ to fail during heat stress. The underlying mechanisms inducing heart failure are still unclear. To pinpoint the processes implicated in HF during heat stress, we measured mitochondrial respiration rates and hydrogen peroxide production of isolated Arctic char (Salvelinus alpinus) heart mitochondria at 4 temperatures: 10°C (acclimation), 15°C, 20°C and 25°C (just over critical maximum). We found that at temperature ranges causing the loss of an organism's general homeostasis (between 20°C and 25°C) and with a substrate combination close to physiological conditions, the heat-induced increase in mitochondrial oxygen consumption levels off. More importantly, at the same state, hydrogen peroxide efflux increased by almost 50%. In addition, we found that individuals with low mitochondrial respiration rates produced more hydrogen peroxide at 10°C, 15°C and 20°C. This could indicate that individuals with cardiac mitochondria having a low respiratory capacity, have a more fragile heart and will be more prone to oxidative stress and HF, and less tolerant to temperature changes and other stressors. Our results show that, at temperatures close to the thermal limit, mitochondrial capacity is compromised and ROS production rates increase. This could potentially alter the performance of the cardiac muscle and lead to heat-induced HF underlining the important role that mitochondria play in setting thermal tolerance limits.

Influences of dietary biotin and avidin on growth, survival, deficiency syndrome and hepatic gene expression of juvenile Nile tilapia Oreochromis niloticus.

This study was undertaken to assess the interactive effects of dietary biotin and avidin on growth, feed conversion, survival and deficiency syndrome of tilapia and to determine the influence of dietary biotin deficiency on the expression of key genes related to biotin metabolism in tilapia. Six iso-nitrogenous and iso-energetic diets based on a common purified basal diet (vitamin-free casein as the protein source) were prepared for this study. The six dietary groups were 0 g avidin with 0 mg biotin (A0B0), 0 g avidin with 0.06 mg biotin/kg diet (A0B1), four avidin-supplemented diets incorporating at a incremental concentrations 0.25, 0.5, 1.0 and 2.0 g/kg diet with 0.06 mg biotin/kg diet (A15B1, A30B1, A60B1 and A120B1). Fish were hand-fed three times a day to apparent satiation for 12 weeks. Each diet was fed to three replicate groups of fish. Fish were kept in glass aquaria in a recirculating aquaculture system under standardized environmental conditions. Growth was significantly higher in fish that received the biotin-supplemented diet (A0B1), compared to diets lacking biotin or supplemented with avidin. Tilapia fed higher concentration of avidin-supplemented diets (A60B1 and A120B1) showed significant growth depression and displayed severe deficiency syndromes such as lethargy, anorexia, circular swimming and convulsions, which ultimately lead to death. There was a strong proportional linear relationship between the avidin content of the diet and feed conversion ratio, FCR (y = 0.43x + 0.135; r = 0.960; P < 0.001) and strong inverse relationship with protein efficiency ratio, PER (y = -0.309x + 2.195; r = 0.961; P < 0.0001). Elevated levels of biotinidase, pyruvate carboxylase, propionyl-CoA carboxylase-A and propionyl-CoA carboxylase-B transcripts were noted in fish fed all graded level of avidin-supplemented diets. A broken-line analysis indicated that feeding tilapia a diet with 44.5 times more avidin than the dietary biotin requirement can induce deficiency syndromes including retarded growth, when analyzing the data of percentage weight gain.

Preliminary evidence of the contribution of the intestinal microflora to biotin supply in zebrafish Danio rerio (Hamilton-Buchanan).

A study was conducted to preliminarily assess the contribution of the intestinal microflora to biotin supply in zebrafish. Biotin and avidin were added to three isonitrogenous and isocaloric purified diets to provide molar avidin: biotin ratios of 0:0 (basal diet), 0:1 (biotin-supplemented diet), and 120:0. Another diet was made by supplementing the antibiotic succinylsulfathiazole (1%, wt/wt) to the basal diet. A fifth diet was the Zeigler commercial diet for zebrafish. Each diet was fed to a triplicate group of fish (mean initial mass 0.266 g) for 8 weeks. The condition factor, feed conversion ratio (FCR), percentage weight gain, and survival were similar in fish groups fed the commercial and the biotin-supplemented diets, but energy conversion efficiency and whole-body biotin content were highest in the fish fed the commercial diet (p<0.05). Reduced growth and survival, and increased FCR were noted in fish fed basal diet compared with those fed biotin-supplemented diet. The supplementation of avidin in diet led to lower survival and condition factor, and higher FCR than that observed with basal diet. Intestinal microbial synthesis is assumed to be a significant source of biotin to the zebrafish, as fish fed the antibiotic-supplemented diet showed the lowest growth, health condition, and feed utilization.

Effects of dietary biotin and avidin on growth, survival, feed conversion, biotin status and gene expression of zebrafish Danio rerio.

A study was conducted to investigate the effects of dietary avidin on growth, survival, food conversion, biotin status and gene expression of zebrafish (Danio rerio Hamilton-Buchanan) juveniles (average wet mass 0.178 g) fed 7 purified diets for 12 weeks. Experimental diets were formulated to provide 0×, 1×, 15×, 30×, 60× and 120× excess avidin versus biotin kg(-1) diet, on a molar basis; a control diet contained neither supplemental biotin nor avidin. Fish fed the control diet had the lowest percentage weight gain and the highest mortality, while the highest percentage weight gain and the lowest mortality was observed with the 0× diet (P<0.05). A linear relationship was observed between feed conversion ratio (FCR) and dietary avidin (r=0.876; P<0.0001). Fish fed diets with 120× more avidin than biotin had the highest whole-body biotin content, while the lowest value was obtained with the control and avidin-free diets (P<0.05). Elevated levels of acetyl CoA carboxylase-A (acca), methylcrotonyl CoA carboxylase (mcc) and propionyl CoA carboxylase-A (pcca) transcripts were recorded in fish fed the control diet, in comparison to the other diets. A broken-line analysis indicated that feeding zebrafish a diet with 60 times more avidin than the dietary biotin requirement level will cause biotin deficiency signs.

Combining suppressive subtractive hybridization and cDNA microarrays to identify dietary phosphorus-responsive genes of the rainbow trout (Oncorhynchus mykiss) kidney.

Phosphorus (P)-responsive genes and how they regulate renal adaptation to phosphorous-deficient diets in animals, including fish, are not well understood. RNA abundance profiling using cDNA microarrays is an efficient approach to study nutrient-gene interactions and identify these dietary P-responsive genes. To test the hypothesis that dietary P-responsive genes are differentially expressed in fish fed varying P levels, rainbow trout were fed a practical high-P diet (R20: 0.96% P) or a low-P diet (R0: 0.38% P) for 7 weeks. The differentially-expressed genes between dietary groups were identified and compared from the kidney by combining suppressive subtractive hybridization (SSH) with cDNA microarray analysis. A number of genes were confirmed by real-time PCR, and correlated with plasma and bone P concentrations. Approximately 54 genes were identified as potential dietary P-responsive after 7 weeks on a diet deficient in P according to cDNA microarray analysis. Of 18 selected genes, 13 genes were confirmed to be P-responsive at 7 weeks by real-time PCR analysis, including: iNOS, cytochrome b, cytochrome c oxidase subunit II , alpha-globin I, beta-globin, ATP synthase, hyperosmotic protein 21, COL1A3, Nkef, NDPK, glucose phosphate isomerase 1, Na+/H+ exchange protein and GDP dissociation inhibitor 2. Many of these dietary P-responsive genes responded in a moderate way (R0/R20 ratio: <2-3 or >0.5) and in a transient manner to dietary P limitation. In summary, renal adaptation to dietary P deficiency in trout involves changes in the expression of several genes, suggesting a profile of metabolic stress, since many of these differentially-expressed candidates are associated with the cellular adaptative responses.

Production of BSA-loaded alginate microcapsules: influence of spray dryer parameters on the microcapsule characteristics and BSA release.

The aim of this study was to optimize the production of BSA-loaded alginate microcapsules by spray drying and to study the release of bovine serum albumin fraction V (BSA) under gastric simulated conditions. Microcapsule yield, BSA release, microcapsule size and size distribution were characterized following the application of different production parameters including inlet air temperature, inlet air pressure and liquid feed rate. The microcapsules were incubated in 0.1 N HCl and BSA release was quantified over time. The yields were higher with the pressure of 3 bar compared to 4 bar and with a feed rate of 0.45 vs. 0.2 ml s(-1). A high feed rate (0.45 vs. 0.2 ml s(-1)) allows one to obtain microcapsules with a low BSA release (p = 0.0327). The increase of the atomizer inlet temperature leads to microcapsules with a higher BSA release (p = 0.0230). A higher air pressure of 4 bar compared to 3 bar resulted in a lower microcapsule size (2.55 vs. 2.80 microm) and led to a narrower size distribution (0.92 vs. 1.07). In conclusion, the spray dryer parameters influenced the alginate microcapsule characteristics as well as subsequent protein release into a simulated gastric medium.

Oral vaccines for finfish: academic theory or commercial reality?

Aquaculture is the fastest growing food-producing sector, providing an acceptable supplement to and substitute for wild fish and plants. Increased production intensification, particularly in high-value species, involves substantial stress, which, as in other captive livestock species, has resulted in outbreaks of major diseases and related mortalities. Widespread use of antibiotics has led to the development of antibiotic-resistant bacteria and the accumulation of antibiotics in the environment and the flesh of fish. Thus, recently effort has been dedicated to vaccine development. Vaccination in fish is complicated by their aquatic environment. Individual injections are labor-intensive and stressful, since fish have to be removed from the water and anaesthetized. Some vaccines offer a limited duration of protection, and thus booster applications are required. In salmonid species, many commercial vaccines use oil-based adjuvants, resulting in a greatly improved duration of protection. However, oil-based adjuvants have been related to significant growth depression, internal adhesions and injection site melanization, resulting in carcass downgrading. Oral administration to aquatic species is by far the most appealing method of vaccine delivery: there is no handling of the fish, which reduces stress; and administration is easy and suitable for mass immunization. However, few oral vaccines have been commercialized, due in part to the increased quantity of antigen required to provoke an immune response, and the lack of an adequate duration of protection. For effective oral delivery, protective antigens must avoid digestive hydrolysis and be taken up in the hindgut in order to induce an effective protective immune response. Antigen encapsulation technologies have been used to protect antigen; however, such strategies can be expensive and are not always effective. Alternative approaches, currently under development, are discussed.