Use of an experimental design to optimise the saponification reaction and the quantification of vitamins A1 and A2 in whole fish.

In ASEAN countries, small freshwater fish species contribute to the nutritional needs of people with few livelihoods by providing them with significant amounts of protein, fat, vitamins and minerals. Some species are eaten whole (with their organs, skin, bones, head and eyes). To estimate the vitamin A content of these foods, conventional saponification has been applied but has not been able to fully release the retinol. Our objective was to optimise the conditions of vitamin A saponification in whole fish to have a reliable estimate of their contribution to intakes. The effects of temperature and saponification time on the retinol quantification of whole fish were evaluated using a two-factor experimental design. Reaction time had a significant effect on the saponification of standard retinyl palmitate and whole fish (p≤0.05). For whole fish, the best conditions for the saponification were to heat the samples to 80 °C for 43 minutes. Under these conditions, the retinol is well liberated from the matrix and protected from degradation and isomerisation reactions. The time-temperature couple used is more intense than that recommended for quantifying vitamin A in milk or enriched margarines. The protective effect of the food matrix against the release of retinol is evident. Vitamin A2 alcohol (3,4-didehydroretinol) was detected in five species and the overall vitamin A contents ranged from 9.6 to 737.5 µg RE/100 g in species frequently consumed in Cambodia. The two species of small fish consumed whole were the ones that contained significantly more vitamin A among the ten tested (p≤0.05). Highlights: Vitamin A2 alcohol was quantified in five fish species. The official saponification partially released retinol in whole fish. The optimised reaction required heating the sample to 80 °C for 43 min.

[The content of zinc and selenium in local food products of Yakutia].

Deficiency of the necessary complex of micronutrients under abnormal nutrition structure creates a problem for the preservation of health and performance of the population. In this regard, it is very relevant to develop a science-based strategy for the consumption of traditional Yakut national food that have a high nutritional value and are able to satisfy the requirements of the human body in micronutrients. The purpose of the research was to investigate the content of selenium and zinc in the local foods which are most commonly consumed by the population of Yakutia. Material and methods. The objects of study were meat (7-9 cuts each) and offal (9-11 species each) of the Yakut cattle breed (2 bulls at the age of 2.5 years), of the Yakut horse foals (3 heads at the age of 6 months), northern domestic deer (3 heads), and whitefish (Coregonus muksun), Yakut crucian carp (Carassius carassius jacuticus), lake minnow [Phoxinus percnurus (Pallas)] (3 kg each). The trace elements, zinc and selenium, were determined by infrared spectroscopy. Results. The greatest amount of zinc in the meat of farm animals was noted in the meat of Yakut cattle (6.8±0.3 mg/100 g) and Yakut horse foals (6.7±0.2 mg/100 g), and the lowest - in the meat of domestic reindeer (1.5±0.1 mg/100 g). In terms of selenium content, the meat of domestic reindeer had the highest indicators (37.0±1.0 µg/100 g), and the meat of Yakut cattle had the lowest (19.0±0.8 µg/100g). The highest concentration of zinc and selenium was found in by-products of reindeer: zinc level in the heart and liver was 12.8 mg/100 g, in the small intestine and rennet - 19.0-20.4 mg/100 g, selenium level in the colon and rennet was 41.0-46.7 µg/100 g. The content of zinc and selenium in the fresh-water muksun belly (2.14±0.08 mg and 45.0±1.8 µg in 100 g) was 32.3-37.2% higher than in the fillet of muksun, and selenium level was 3 fold higher than in the Yakut carp and in the lake minnow. The consumption of 100-200 g of meat or by-products of Yakut cattle, Yakut horse foals, by-products of reindeer or Yakut crucian carp can fully cover the daily requirement of an adult in zinc. The consumption of 200 g of venison or muksun completely covers the daily need for selenium, while the portion of the other studied products contains about half or more of recommended daily intake of this trace element. Conclusion. The data presented in the article show that the population of Yakutia, with a rational diet with local products, can meet the requirements in selenium and zinc in accordance with physiological needs.

Comparison of the Fatty Acid and Triglyceride Profiles of Big Eye Tuna (Thunnus obesus), Atlantic salmon (Salmo salar) and Bighead Carp (Aristichthysnobilis) Heads.

Big eye tuna (Thunnus obesus), Atlantic salmon (Salmo salar) and bighead carp (Aristichthys nobilis) are three representative marine and fresh water fishes. In this study, the content of total lipids (TL), triglyceride (TG) fraction, and the fatty acid profiles in the corresponding fish heads were analyzed. Meanwhile, their complicated TG molecular species were further characterized. The results showed that TG was the major lipid in these three fish heads (60.58-86.69%). Compared with other two fish heads, big eye tuna head was the most abundant in polyunsaturated fatty acids, among which eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) accounted for 64.29% and 32.77% in the TL and TG fraction, respectively. It is also worth noting that EPA+DHA/total fatty acid (TFA) value of TL and TG fraction from bighead carp head showed no significant difference with Atlantic salmon head, a typical marine fish. There were 146 TG molecules detected in big eye tuna head, 90 in Atlantic salmon and 87 in bighead carp heads. DHA or EPA accounted for 56.12%, 22.88%, and 5.46% of the total TG molecules in these three fish heads, respectively. According to principal component analysis, orthogonal projection to latent structures-discriminant analysis and the constructed heat map, the three samples could be completely differentiated based on their TG molecule fingerprints. This study is the first to compare marine and fresh water fish from the perspective of their heads' fatty acid and TG molecule profiles.

Myxobolus sp. and Henneguya sp. (Cnidaria: Myxobolidae) natural co-infection in the kidney of Piaractus mesopotamicus (Characiformes: Serrasalmidae).

This study evaluated the myxozoan infection and histopathology of the kidney of freshwater fish Piaractus mesopotamicus from intensive fish farming in Brazil. A total of 55 fish were examined for myxozoan infection. Infected organs were processed by usual histology and stained with hematoxylin-eosin (H&E) and Ziehl-Neelsen (ZN). From the total of 55 fish analyzed, 47 (85.45%) presented myxospores, being 9.09% (5/55) only with Myxobolus sp., 5.45% (3/55) only with Henneguya sp., and 70.91% (39/55) presenting both parasites. The presence of myxospores was associated with histological alterations in both stromal and renal parenchyma. Myxospores were found mostly in the peritubular interstitial tissue and in low intensity in the glomerulus which caused nuclear hypertrophy and loss of Bowman space. An increase in the glomerular tuft and a reduction in the lumen of the collector tubules were also observed, besides the high number of melanomacrophage cells in the glomerulus. This study reports for the first time detection of myxozoan mixed infection in one organ of pacu and discuss the possible transportation of myxospores in the circulating blood.

Water deprivation-induced hypoxia and oxidative stress physiology responses in respiratory organs of the Indian stinging fish in near coastal zones.

Background: Water deprivation-induced hypoxia stress (WDIHS) has been extensively investigated in numerous fish species due to their adaptation with accessory respiratory organs to respire air but this has not been studied in Indian stinging fish Heteropneustes fossilis. Data regarding WDIHS-induced metabolism in accessory respiratory organ (ARO) and gills and its relationship with oxidative stress (OS) in respiratory organs of air-breathing fish H. fossilis, are limited. So, this study aimed to investigate the effects of WDIHS (0, 3, 6, 12, and 18 h) on hydrogen peroxide (H2O2) as reactive oxygen species (ROS), OS, redox regulatory enzymes, and electron transport enzymes (ETC) in ARO and gills of H. fossilis. Methods: Fish were exposed to air for different hours (up to 18 h) against an appropriate control, and ARO and gills were sampled. The levels of oxygen saturation in the body of the fish were assessed at various intervals during exposure to air. Protein carbonylation (PC) and thiobarbituric acid reactive substances (TBARS) were used as OS markers, H2O2 as ROS marker, and various enzymatic activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), along with the assessment of complex enzymes (I, II, III, and V) as well as the levels of ascorbic acid (AA) and the reduced glutathione (GSH) were quantified in both the tissues. Results: Discriminant function analyses indicate a clear separation of the variables as a function of the studied parameters. The gills exhibited higher levels of GSH and H2O2 compared to ARO, while ARO showed elevated levels of PC, TBARS, AA, SOD, CAT, and GPx activities compared to the gills. The activities of GR and ETC enzymes exhibited similar levels in both the respiratory organs, namely the gills, and ARO. These organs experienced OS due to increased H2O2, TBARS, and PC levels, as observed during WDIHS. Under WDIHS conditions, the activity/level of CAT, GPx, GR, and GSH decreased in ARO, while SOD activity, along with GR, GSH, and AA levels decreased in gills. However, the activity/level of SOD and AA in ARO and CAT in gills was elevated under WDIHS. Complex II exhibited a positive correlation with WDIHS, while the other ETC enzymes (complex I, III, and V) activities had negative correlations with the WDIHS. Discussion: The finding suggests that ARO is more susceptible to OS than gills under WDIHS. Despite both organs employ distinct redox regulatory systems to counteract this stress, their effectiveness is hampered by the inadequacy of small redox regulatory molecules and the compromised activity of the ETC, impeding their ability to effectively alleviate the stress induced by the water-deprivation condition.

Exploring the Relationships Between Bacterial Community, Taste-Enhancing Peptides and Aroma in Thai Fermented Fish (Pla-ra).

As a traditional Thai condiment, Pla-ra is used to add flavor and richness to dishes. Nine treatment combinations of Pla-ra formulations created from 3 types of fish (Mor fish, Kradee fish, and Mor + Kradee fish) and 4 different carbohydrate sources (none, rice bran, roasted rice, and rice bran─roasted rice mixture) were studied through a 12 month fermentation period (1, 3, 5, 7, 8, 9, 10, 11, and 12 months). 16S rRNA Next Generation Sequencing (NGS) and LC-MS/MS techniques were used to analyze the microbial diversity and identify taste-enhancing peptides. Descriptive sensory analysis was performed on the extracts of the 108 Pla-ra samples mixed in a model broth. Koku perception and saltiness-enhancing attributes were clearly perceived and dominant in all samples, even though glutamyl peptides, including γ-Glu-Val-Gly, were found at subthreshold levels. The samples from mixed fish and Mor fish fermented with roasted ground rice and rice bran for 12 months had the most typical Pla-ra odors and tastes and had high taste-enhancing activities. NGS analysis revealed the presence of bacteria containing a large number of protease and aminopeptidase genes in the samples. Bacillus spp., Gallicola spp., and Proteiniclasticum spp. correlated well with the generation of glutamyl and arginyl peptides and typical odors in the samples. These results confirmed the typical sensory quality of Pla-ra depended on protein sources, carbohydrate sources, and bacteria communities. Further optimization of the microbial composition found could lead to the development of starter cultures to control and promote flavor development in fermented fish products.

Dietary Macroalgae Saccharina japonica Ameliorates Liver Injury Induced by a High-Carbohydrate Diet in Swamp Eel (Monopterus albus).

A high-carbohydrate diet lowers the rearing cost and decreases the ammonia emission into the environment, whereas it can induce liver injury, which can reduce harvest yields and generate economic losses in reared fish species. Macroalgae Saccharina japonica (SJ) has been reported to improve anti-diabetic, but the protective mechanism of dietary SJ against liver injury in fish fed a high-carbohydrate diet has not been studied. Therefore, a 56-day nutritional trial was designed for swamp eel Monopterus albus, which was fed with the normal diet [20% carbohydrate, normal carbohydrate (NC)], a high carbohydrate diet (32% carbohydrate, HC), and a HC diet supplemented with 2.5% SJ (HC-S). The HC diet promoted growth and lowered feed coefficient (FC), whereas it increased hepatosomatic index (HSI) when compared with the NC diet in this study. However, SJ supplementation increased iodine contents in muscle, reduced HSI, and improved liver injury, such as the decrease of glucose (GLU), total bile acid (TBA), and alanine aminotransferase (ALT) in serum, and glycogen and TBA in the liver. Consistently, histological analysis showed that SJ reduced the area of lipid droplet, glycogen, and collagen fiber in the liver (p < 0.05). Thoroughly, the underlying protective mechanisms of SJ supplementation against HC-induced liver injury were studied by liver transcriptome sequencing coupled with pathway analysis. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the differentially expressed genes (DEGs), such as the acetyl-coenzyme A synthetase (acss1), alcohol dehydrogenase (adh), interferon-induced protein with tetratricopeptide repeats 1 (ifit1), aldo-keto reductase family 1 member D1 (akr1d1), cholesterol 7-alpha-monooxygenase (cyp7a1), and UDP-glucuronosyltransferase (ugt), indicated that the pathway of glycolysis/gluconeogenesis was the main metabolic pathway altered in the HC group compared with the NC group. Meanwhile, hepatitis C, primary BA biosynthesis, and drug metabolism-cytochrome P450 were the three main metabolic pathways altered by SJ supplementation when compared with the HC group. Moreover, the BA-targeted metabolomic analysis of the serum BA found that SJ supplementation decreased the contents of taurohyocholic acid (THCA), taurochenodeoxycholic acid (TCDCA), taurolithocholic acid (TLCA), nordeoxycholic acid (NorDCA), and increased the contents of ursocholic acid (UCA), allocholic acid (ACA), and chenodeoxycholic acid (CDCA). In particular, the higher contents of UCA, ACA, and CDCA regulated by SJ were associated with lower liver injury. Overall, these results indicate that the 2.5% supplementation of SJ can be recommended as a functional feed additive for the alleviation of liver injury in swamp eel-fed high-carbohydrate diets.

A phylogeny of the genus Limia (Teleostei: Poeciliidae) suggests a single-lake radiation nested in a Caribbean-wide allopatric speciation scenario.

OBJECTIVE: The Caribbean is an important global biodiversity hotspot. Adaptive radiations there lead to many speciation events within a limited period and hence are particularly prominent biodiversity generators. A prime example are freshwater fish of the genus Limia, endemic to the Greater Antilles. Within Hispaniola, nine species have been described from a single isolated site, Lake Miragoâne, pointing towards extraordinary sympatric speciation. This study examines the evolutionary history of the Limia species in Lake Miragoâne, relative to their congeners throughout the Caribbean. RESULTS: For 12 Limia species, we obtained almost complete sequences of the mitochondrial cytochrome b gene, a well-established marker for lower-level taxonomic relationships. We included sequences of six further Limia species from GenBank (total N = 18 species). Our phylogenies are in concordance with other published phylogenies of Limia. There is strong support that the species found in Lake Miragoâne in Haiti are monophyletic, confirming a recent local radiation. Within Lake Miragoâne, speciation is likely extremely recent, leading to incomplete lineage sorting in the mtDNA. Future studies using multiple unlinked genetic markers are needed to disentangle the relationships within the Lake Miragoâne clade.

Replacing fish meal by food waste to produce lower trophic level fish containing acceptable levels of polycyclic aromatic hydrocarbons: Health risk assessments.

This study aimed at using different types of food wastes (mainly containing cereal [food waste A] and meat meal [food waste B]) as major sources of protein to replace the fish meal used in fish feeds to produce quality fish. The traditional fish farming model used to culture low trophic level fish included: bighead, (Hypophthalmichthys nobilis), grass carp, (Ctenopharyngodon idellus), and mud carp, (Cirrhinus molitorella) of omnivorous chain. The results indicated that grass carp and bighead carp fed with food waste feeds were relatively free of PAHs. The results of health risk assessment showed that the fish fed with food waste feeds were safe for consumption from the PAHs perspective.

Replacing fish meal by food waste in feed pellets to culture lower trophic level fish containing acceptable levels of organochlorine pesticides: health risk assessments.

The present study used food waste (collected from local hotels and restaurants) feed pellets in polyculture of low-trophic level fish [bighead (Aristichtys nobilis), grass carp (Ctenopharyngodon idellus), and mud carp (Cirrhina molitorella)] aiming at producing safe and quality products for local consumption. The results indicated that grass carp (hexachlorocyclohexanes (HCHs) <0.03; dichlorodiphenyltrichloroethanes (DDTs) 1.42-3.34 ng/g ww) and bighead carp (HCHs<0.03; DDTs 1.55-2.56 ng/g ww) fed with food waste feed pellets were relatively free of organochlorine pesticides (OCPs). The experimental ponds (water and sediment) were relatively free of OCPs, lowering the possibility of biomagnification of OCPs in the food chains within the ponds. The raw concentrations of OCPs extracted from the fish were not in the bioavailable form, which would ultimately reach bloodstream and exert adverse effects on human body. Health risk assessments based on digestible concentrations are commonly regarded as a more accurate method. The results of health risk assessments based on raw and digestible concentrations showed that the fish fed with food waste feed pellets were safe for consumption from the OCP perspective.

Validade comercial de filés de Tilápia do Nilo (Oreochromis niloticus) resfriados embalados em atmosfera modificada e irradiados / Shelf life of refrigerated tilapia fillets (Oreochromis niloticus) packed in modified atmosphere and irradiated

O objetivo do presente estudo foi avaliar os efeitos dos processos de irradiação (com dose de 1,5kGy) e embalagem em atmosfera modificada (60% N2 e 40% CO2), aplicados isoladamente ou combinados, na extensão da validade comercial de filés de Tilápia do Nilo (Oreochromis niloticus) resfriados. Nos experimentos, foram utilizados 120 filés separados em lotes, de acordo com o tratamento: (T1) embalagem em ar (controle), (T2) embalagem em atmosfera modificada, (T3) embalagem em ar e irradiação, (T4) embalagem em atmosfera modificada e irradiação. Foram realizadas análises físico-químicas (BVT, TMA, pH, amônia e TBARS), bacteriológicas (contagem de bactérias heterotróficas, aeróbias, mesófilas e psicrotróficas) e avaliação sensorial. Os resultados consistentemente indicaram uma redução progressiva da qualidade do produto com o tempo de armazenamento, a qual foi significativamente mais rápida nas amostras controle do que naquelas submetidas aos demais tratamentos, tendo sido observada uma extensão na validade comercial dos filés embalados em atmosfera modificada e/ou irradiados de 4 dias para aproximadamente duas semanas.

This paper investigated the effects of modified atmosphere packing (60% N2 and 40% CO2) and irradiation (with a 1.5kGy dose) either used alone or combined on the shelf life of fillets of refrigerated Nile Tilapia. A total of 120 fillets were used in the experiments in order to test four different treatments and included samples: packed in air (control), packed in modified atmosphere, packed in air and irradiated, and finally, packed in modified atmosphere and irradiated. Regular monitoring of physicochemical parameters (TVB, pH, ammonia and TBARS), bacteriological conditions (heterotrophic aerobic mesophilic and psychrotrophic bacteria) and sensory acceptance were performed. The results consistently indicated an increasing deterioration of the fillets with time, however those treated with irradiation, modified atmosphere or both combined had longer shelf lives (around two weeks) in comparison with the control (4 days only).