Exploring fishery waste potential as antifouling component.

Marine biofouling is a global issue with economic and ecological implications. Existing solutions, such as biocide-based antifouling paints, are toxic for the environment. The search for better antifouling agents remains crucial. Recent research focuses on eco-friendly antifouling paints containing natural compounds like enzymes. This study evaluates enzymatic extracts from fishery residues for antifouling potential. Extracts from Pleoticus muelleri shrimp, Illex argentinus squid, and Lithodes santolla king crab were analyzed. Proteolytic activity and thermal stability were assessed, followed by bioassays on mussel byssus thread formation and barnacle cypris adhesive footprints. All three extracts demonstrated proteolytic activity and 24-h stability at temperate oceanic temperatures, except I. argentinus. P. muelleri extracts hindered cyprid footprint formation and mussel byssus thread generation. Further purification is required for L. santolla extract to assess its antifouling potential activity. This study introduces the use of fishery waste-derived enzyme extracts as a novel antifouling agent, providing a sustainable tool to fight against biofouling formation.

Understanding the Digestive Peptidases from Crustaceans: from Their Biochemical Basis and Classical Perspective to the Biotechnological Approach.

Scientific studies about decapod crustaceans' digestive physiology have increased, being an important topic with novel results in the last years. This revision aims to show how the study of crustacean peptidases has evolved, from the classical biochemical characterization studies to the assessment of their usefulness in biotechnological and industrial processes, with emphasis on commercial species of interest to world aquaculture and fisheries. First studies determined the proteolytic activity of the midgut gland crude extracts and evaluated the optimum biochemical properties of specific enzymes. Peptidase's identity was determined using inhibitors and specific protein substrates on tube tests and electrophoresis gels. Later, various studies focused on the characterization of purified peptidases and their gene expression. Recently, the integrated mechanisms of enzyme participation during the digestive process of food protein have been established using novel techniques. Scientific research has revealed some of the potential biotechnological applications of crustacean peptidases in the food industry and other processes. However, the knowledge field is enormous, and there is much to explore and study in the coming years.

Digestive proteinases from the marine fish processing wastes of the South-West Atlantic Ocean: Their partial characterization and comparison.

Fish processing generates plenty of waste that is directly discarded in open-air dumps and water sources, or treated in the same way as urban solid waste, causing serious pollution problems. The waste represents a significant source of high-value bioproducts with potential applications in different industrial processes such as the production of feed, fertilizers, biodiesel and biogas, detergent additives and cosmetics. The objective of this study was to characterize and compare specific activities under different pH values and temperature conditions of acid and alkaline proteinases and viscera yield from the following fish species: Argentine hake Merluccius hubbsi, Brazilian flathead Percophis brasiliensis, Brazilian codling Urophycis brasiliensis and Stripped weakfish Cynoscion guatucupa. Individuals were fished off the coast of Mar del Plata (Argentina) by a commercial fleet and the viscera were immediately extracted and kept on ice until use. Stomach proteinases from four species had the highest activity at pH 2, with stability in the range of pH 2-4. The optimum pH was 11.5 from intestinal enzymes of C. guatucupa, M. hubbsi and P. brasiliensis and 9.5 from intestinal enzymes of U. brasiliensis. Alkaline proteinases from all species were highly stable in the range of 7-11.5. The optimum temperature of stomach proteinases from the four species studied were 30 and 50°C, with stability at 10 and 30°C during 150 min. The optimum temperature of intestinal enzymes from the tested species were 50°C with high stability at 10 and 30°C during 150 min. Alkaline proteinase from all species and acid proteinases from C. guatucupa were inactive at 70°C after 150 min, while there was a residual activity lower than 5% at 80°C on pre-incubated stomach enzymes of M.hubbsi, P. brasiliensis and U. brasiliensis after 5, 10 and 20 min, respectively. Digestive proteinases recovered in this study could be appropriate for technological usage, reducing manufacturing costs, obtaining revenue from fishery wastes, and contributing to the reduction of environmental pollution.

De novo assembly and transcriptome characterization of the freshwater prawn Palaemonetes argentinus: Implications for a detoxification response.

Palaemonetes argentinus, an abundant freshwater prawn species in the northern and central region of Argentina, has been used as a bioindicator of environmental pollutants as it displays a very high sensitivity to pollutants exposure. Despite their extraordinary ecological relevance, a lack of genomic information has hindered a more thorough understanding of the molecular mechanisms potentially involved in detoxification processes of this species. Thus, transcriptomic profiling studies represent a promising approach to overcome the limitations imposed by the lack of extensive genomic resources for P. argentinus, and may improve the understanding of its physiological and molecular response triggered by pollutants. This work represents the first comprehensive transcriptome-based characterization of the non-model species P. argentinus to generate functional genomic annotations and provides valuable resources for future genetic studies. Trinity de novo assembly consisted of 24,738 transcripts with high representation of detoxification (phase I and II), anti-oxidation, osmoregulation pathways and DNA replication and bioenergetics. This crustacean transcriptome provides valuable molecular information about detoxification and biochemical processes that could be applied as biomarkers in further ecotoxicology studies.

Energy reserves mobilization: Strategies of three decapod species.

In food deprivation assays, several different responses have been observed in crustaceans. However, studying energy reserves utilization among more than one species during the same starvation period has not yet been performed, particularly to discern whether the responses are due to intrinsic and/or environmental factors. We hypothesize that decapod species with similar feeding habits have the same strategies in the use of energetic reserves during starvation, even though they inhabit different environments. The aim of this study was to compare the energy reserves mobilization of three decapods species (Cherax quadricarinatus, Palaemon argentinus and Munida gregaria) with similar feeding habits, exposed to similar food deprivation conditions. The crayfish, shrimp and squat-lobster were experimentally kept at continuous feeding or continuous starvation throughout 15 days. Every 3rd day, the midgut gland index (MGI), and the glycogen, lipid and protein contents were measured in the midgut gland (MG) and pleon muscle. Palaemon argentinus mobilized more reserves during starvation, followed by C. quadricarinatus, and the last M. gregaria. The starved shrimps presented low MGI, whereas MG showed a reduction in glycogen (from day 6 to 15), lipid (from day 3 to 15), and protein levels (at day 9 and 15) while in their muscle, lipid reserves decreased at days 3 and 6. In C. quadricarinatus, the most affected parameters in the MG were MGI, glycogen (from day 6 to 15), and lipids (at day 12 and 15). In the MG of M. gregaria only the glycogen was reduced during fasting from 3 to 15 days. Even though the three studied species have similar feeding habitats, we found that their energetic profile utilization is different and it could be explained by the habitat, life span, temperature, organ/tissue, and metabolism of the species. Our results may be useful to understand the several different responses of crustaceans during starvation.

Liposoluble vitamins in Crustacean feed: Metabolic and Histological responses.

Vitamins are vital for normal growth and survival of living organisms and they are distributed in feedstuffs in small quantities. This review is focused on the liposoluble vitamins (A, D, E and K) in the diets and metabolic responses of the Argentine penaeoid shrimps Pleoticus muelleri and Artemesia longinaris, distributed along the South American coast line. Growth, survival and histological analyses serve as indicators of the nutritional value derived from vitamin deficiency. Liposoluble vitamins are also related to stress, antioxidant defense and immune response of shrimps. Effective diet for shrimp culture that provide not only macronutrients including protein and lipid but also micronutrients such as vitamins for optimal growth is an ever improving subject. This review may help formulating suitable feeds for shrimps.

Are Mussels Always the Best Bioindicators? Comparative Study on Biochemical Responses of Three Marine Invertebrate Species to Chronic Port Pollution.

Bivalves have traditionally been considered good bioindicators due to their sensitivity to pollution, among other features. This characteristic is shared by several other non-bivalve species as well, though studies in this respect remain scarce. This work aims to compare biomarker sensitivity to chronic port pollution among three intertidal invertebrate species with good bioindicator characteristics. Mussels' immunological (phenoloxidase and peroxidases) and biotransformation (glutathione-S-transferase) responses were contrasted against those of limpets and barnacles. The three species under study evidenced activity of all the enzymes measured, although with differences. Barnacle Balanus glandula was the most sensitive species showing pollution modulation of the three enzymes, which suggests that mussels would not always be the best bioindicator species among marine invertebrates depending on the responses that are assessed.

Long-Term Starvation and Posterior Feeding Effects on Biochemical and Physiological Responses of Midgut Gland of Cherax quadricarinatus Juveniles (Parastacidae).

We investigated the effect of long-term starvation and posterior feeding on energetic reserves, oxidative stress, digestive enzymes, and histology of C. quadricarinatus midgut gland. The crayfish (6.27 g) were randomly assigned to one of three feeding protocols: continuous feeding throughout 80 day, continuous starvation until 80 day, and continuous starvation throughout 50 day and then feeding for the following 30 days. Juveniles from each protocol were weighed, and sacrificed at day 15, 30, 50 or 80. The lipids, glycogen, reduced glutathione (GSH), soluble protein, lipid peroxidation (TBARS), protein oxidation (PO), catalase (CAT), lipase and proteinase activities, and histology were measured on midgut gland. Starved crayfish had a lower hepatosomatic index, number of molts, specific growth rate, lipids, glycogen, and GSH levels than fed animals at all assay times. The starvation did not affect the soluble protein, TBARS, PO levels and CAT. In starved juveniles the lipase activity decreased as starvation time increased, whereas proteinase activity decreased only at day 80. The histological analysis of the starved animals showed several signs of structural alterations. After 30 days of feeding, the starved-feeding animals exhibited a striking recovery of hepatosomatic index, number of molts, lipids and glycogen, GSH, lipase activity and midgut gland structure.

The shrimp mitochondrial FoF1-ATPase inhibitory factor 1 (IF1).

The whiteleg shrimp species Litopenaeus vannamei is exposed to cyclic changes of the dissolved oxygen concentration of seawater and must neutralize the adverse effects of hypoxia by using ATP as energy source. In crustaceans, the mitochondrial FOF1-ATP synthase is pivotal to the homeostasis of ATP and function prevalently as a FOF1-ATPase. Hitherto, it is unknown whether these marine invertebrates are equipped with molecules able to control the FOF1-ATPase inhibiting the ATP consumption. In this study, we report two variants of the mitochondrial FOF1-ATPase Inhibitory Factor 1 (IF1) ubiquitously expressed across tissues of the Litopenaeus vannamei transcriptome: the IF1_Lv1 and the IF1_Lv2. The IF1_Lv1, with a full-length sequence of 550 bp, encodes a 104 aa long protein and its mRNA amounts are significantly affected by hypoxia and re-oxygenation. The IF1_Lv2, with a sequence of 654 bp, encodes instead for a protein of 85 aa. Both proteins share a 69 % homology and contain a conserved minimal inhibitory sequence (IATP domain) along with a G-rich region on their N-terminus typical of the invertebrate. In light of this characterization IF1 is here discussed as an adaptive mechanism evolved by this marine species to inhibit the FOF1-ATPase activity and avoid ATP dissipation to thrive in spite of the changes in oxygen tension.

In vivo and in vitro protein digestibility of formulated feeds for Artemesia longinaris (Crustacea, Penaeidae)

This study was undertaken to determine the in vivo crude protein apparent digestibility in the prawn Artemesia longinaris, using feeds with 0.25 percent of chromic oxide and animal (fish meal, meat and bone meal and squid protein concentrate) and plant (soybean meal) ingredients. Three replicate groups of prawn were fed and the feces were collected. The rate of protein hydrolysis was measured in vitro using midgut gland enzyme extract from the prawns fed the respective feeds and was compared with those found with enzyme extract of wild prawn. The in vivo apparent digestibility coefficients showed significant differences among the feeds (P<0.05). Fish meal feed presented the highest digestibility (92 percent); intermediate digestibility (83 percent) was found for meat and bone meal feed, and the less digestible feed (63 percent) was that containing soybean meal and squid proteins concentrate. No significant differences in the in vitro protein digestibility were found among the experimental feeds. The results indicated the limitation of in vitro enzyme assays and that it should be complemented by in vivo studies.

O objetivo do presente trabalho foi determinar a digestibilidade aparente in vivo da proteína bruta de ingredientes de origem animal (farinhas de peixe, osso e carne e concentrado de proteína de lula) e ingredientes vegetais (farinha de soja) em camarões Artemesia longinaris utilizando rações contendo 0,25 por cento de óxido de cromo. Três grupos de camarões, utilizados como replicatas, foram alimentados e as fezes coletadas. A velocidade de hidrólise da proteína de cada ração foi medida in vitro utilizando extrato enzimático da glândula do intestino médio dos camarões alimentados com a ração correspondente e foi comparado com aqueles obtidos com o extrato enzimático de camarões selvagens. Os coeficientes de digestibilidade aparente in vivo mostraram diferenças significativas entre as rações testadas (P<0,05). A farinha de peixe apresentou a maior digestibilidade (92 por cento), enquanto valores intermediários de digestibilidade (83 por cento) foram encontrados para a farinha de carne e ossos. A ração contendo farinha de soja e concentrado de proteína de lula resultou em menor digestibilidade (63 por cento). Não houve diferença significativa entre os valores de digestibilidade in vitro para as rações testadas. Estes resultados indicam a limitação inerente dos ensaios enzimáticos in vitro, os quais poderiam ser complementados com estudos in vivo.

Antioxidant activity in hepatopancreas of the shrimp (Pleoticus muelleri) by electron paramagnetic spin resonance spectrometry.

Free radical scavenging properties of hepatopancreas extracts of Pleoticus muelleri were evaluated by electron paramagnetic spin resonance spectrometry methods (EPR) against the stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. The present study was carried out to characterize different physiological stages of the shrimp under environmental and nutritional stress, evaluating the effect on growth, survival, and functional morphology of the hepatopancreas. Feeding trials were carried out on juveniles (1 g initial weight) held in aquaria. Each diet, with different concentrations of vitamins A and E, was tested in triplicate groups during 25 days. The control groups were fed with fresh squid mantle and with a vitamin-free diet. For all of the diets, the extracts exhibited strong DPPH radical scavenging activity, suggesting that the tissue is a powerful natural antioxidant. Individuals fed with different concentrations of vitamin E showed the strongest effect on the DPPH radicals, reducing the DPPH radicals to 50%, after an incubation period of 3 min. In contrast, the extracts of control animals, fed with squid mantle, had the weakest antioxidant activity (4%). These data indicated that the presence of vitamin E in the diet can provide immediate protection against free radicals.