Supercritical CO2 Fluid Extraction and Microencapsulation of Oil from Anchovy (Engraulis mordax) By-Products.

Research background: Fish by-products are discarded as waste, which has a significant impact on the environment. They have no economic value, but there are many opportunities to turn them into high value products. Due to significant quantities generated internationally and the continuous expansion of the market for ω-3 and ω-6 fatty acids as nutraceuticals, innovative technological approaches are needed to transform this waste into marketable products with added value, while limiting the risk of environmental pollution. Experimental approach: In this study, two temperatures (40 and 60 °C) at a constant pressure during the extraction of anchovy by-products with supercritical CO2 fluid were used to determine extraction yield, fatty acid, tocopherol and phytosterol composition, followed by microencapsulation with two matrices based on the transglutaminase-mediated crosslinking reaction between whey protein isolates and casein. Before microencapsulation, the binding parameters were estimated using quenching studies. Results and conclusions: The results showed a higher content of total fatty acids when extracted at 40 °C, resulting in two fractions on a dry mass basis of (712±12) mg/g in the fraction obtained in the separator with code S40 and (732±10) mg/g in the fraction obtained in the separator with code S45, respectively. The monounsaturated (MUFAs) and polyunsaturated fatty acids (PUFAs) accounted for 40-44 %. The extracts showed a higher mass fraction of eicosapentaenoic acid ((28.7±1.0) mg/g) in fraction S45 when extracted at 60 °C. A minimum inhibitory and bactericidal concentration of 0.66 µg/mL against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923 was found for all fractions. Higher binding constants were found for palmitoleic and oleic acids than for palmitic acid. The control variant, without crosslinking, enabled the microencapsulation of a higher amount of fatty acids, while in both powders the sum of MUFAs and PUFAs was 40 %. Novelty and scientific contribution: The approaches used in our study open up new opportunities for adding value to the fish by-products through extraction and microencapsulation, extending their potential use to food, cosmetics and nutraceuticals.

Accidental cannulation of amoebic liver abscess during pericardiocentesis: a case report.

Background: Amoebiasis is a prevalent infection in the tropics and can sometimes present as liver abscess. Cardiac tamponade is an uncommon complication of ruptured amoebic liver abscess requiring urgent pericardiocentesis, which has a high success rate, but procedural complications can include injury to cardiac chambers, abdominal viscera, and even death. This case underscores the approach to diagnose and manage an unintended visceral puncture during pericardiocentesis, which is a rare but life-threatening complication. Case summary: A 41-year-old male presented with intermittent fever over 2 months and chest pain for 15 days. Echocardiography revealed a significant pericardial effusion causing cardiac tamponade. In an emergency setting, percutaneous pericardiocentesis was attempted to drain the effusion. However, the pigtail inadvertently punctured a sizable liver abscess. Consequently, another pigtail was inserted into the pericardial cavity to successfully drain the effusion. Patient was discharged on Day 12 and is doing well at 6 months follow-up. Discussion: A previously undiagnosed case of a ruptured amoebic liver abscess presented with the uncommon complication of cardiac tamponade, necessitating emergency pericardiocentesis, which inadvertently led to the cannulation of the liver abscess. This case underscores the significance of image-guided pericardiocentesis in minimizing procedural complications. This case also highlights the intricacies of addressing accidental visceral puncture during pericardiocentesis, specially involving the liver. It also underscores the need to consider the possibility of a ruptured amoebic liver abscess when anchovy sauce-like pus is drained from pericardial cavity, especially in high epidemiologically prevalent country like India.

Effects of diet and habitat on Hg levels in Japanese anchovy in the high seas of the northwestern Pacific Ocean.

The aim of this study was to explore the effects of diet and habitat on the Hg levels of Japanese anchovy at different growth stages. We measured the amounts of Hg and stable carbon and nitrogen isotopes in the muscle and stable carbon and oxygen isotopes in the otoliths of 54 Japanese anchovy specimens obtained from the open seas of the Northwest Pacific Ocean (39°2'N âˆ¼ 42°30'N, 154°02'E âˆ¼ 161°29'E) between June and July 2021. Body length had a significant effect on δ13Cbulk, δ15Nbulk and Moto (P < 0.01). As individuals grew, δ13Cbulk and δ15Nbulk tended to gradually increase, but Moto tended to gradually decrease. The variation in the niches of Japanese anchovy at different growth stages showed that the standard ellipse-corrected area subsequently decreased with growth. It reached its smallest value in the 136-150 mm group, and there was no overlap between the 136-150 mm group and the other groups. The GAM results showed that Hg levels tended to decrease first and then increase with growth. There was a positive correlation between Hg levels and δ18Ooto in fish. Hg levels increased gradually with increasing δ15Nbulk. In our study, there may be a gradual shift in the diet of Japanese anchovy from phytoplankton to prey at higher trophic levels, and the depth of seawater in which the predators feed gradually increased with growth. Changes in diet and habitat were probably the main reasons for the increase in Hg levels.

The impact of raw material form and starter culture inoculation on bacterial communities and metabolites in fermented anchovy sauce.

This study explored the dynamics of anchovy sauce fermentation and investigated how the raw material form and the use of starter cultures affect bacterial and metabolite profiles. Using a comprehensive approach, we examined the fermentation process using anchovies in two forms (whole and ground) and three different starter cultures. The use of ground anchovies resulted in an accelerated fermentation process for anchovy sauce; however, the increased diversity of bacterial phylotypes and altered accumulation of biogenic amines were observed. Inoculation of starter cultures resulted in a shift from spontaneous to controlled fermentation, highlighting their ability to regulate bacterial communities. Despite a slightly reduced fermentation rate, inoculation with Tetragenococcus halophilus was shown to be a potent method for reducing biogenic amines and affecting metabolite profiles. As the industry strives to balance fermentation speed and quality, our research could provide insights for improving the efficiency, safety, and quality of anchovy sauce production.

Anchovy and sardine condition and energy content in the North Aegean Sea (eastern Mediterranean) in relation to their contrasting reproductive strategies.

Forage species with high biomass, such as anchovies and sardines, play a key role in pelagic ecosystems and make up a significant proportion of the world's capture fisheries production. In recent years, condition indices have gained interest as significant indicators for assessing the effects of environmental and human pressures on these species and the quality of their habitats. In the present study, we examined, for the first time in the North Aegean Sea (eastern Mediterranean), the year-round variation in somatic and gonadal condition, energy density, and percentage of lipid content of anchovy (Engraulis encrasicolus) and sardine (Sardina pilchardus). Energy density was measured with bomb calorimetry and percentage lipid content with the fatmeter, a portable electronic device. Finally, the monthly changes in gonadal and energetic condition were examined in relation to the annual cycle of temperature and mesozooplankton biomass, simulated by the implementation of a coupled hydrodynamic-biogeochemical model (POM-ERSEM). There was a strong relationship between fish energy density (kJ g-1) and percentage dry weight. Furthermore, the mean monthly energy density and fatmeter measurements were strongly correlated, especially in sardine. Overall, the monthly changes in energetic condition were indicative of the species' different strategies for energy acquisition and allocation to reproduction (capital vs. income breeding): sardine exhibited low energy density and percentage lipid content during the winter spawning period (November-March) and markedly higher energetic condition from spring to autumn (April-October). Anchovy spawning period, as inferred from gonadal condition, lasted from April to September, i.e., during the warm period of the year but its energy density and percentage lipid content did not exhibit any seasonal changes and were markedly lower than in sardine from April to October. Finally, the simulated mesozooplankton biomass was higher from January to July, which corresponded to the second half of the spawning season for sardine, but first half of the spawning season for anchovy.

The performance of anchovy purse seine in the North Coastal Java Sea, Indonesia.

Small pelagic fishes including anchovies in Indonesia are caught by using different types of fishing gears. However, the anchovy purse seine commonly used polyethylene (PE) as material webbing which is less effective in the North Coastal Java Sea. This study aims to describe the design and construction of the anchovy purse seine and the characteristics of this gear construction. The research was carried out in the North Coastal Java Sea of Indonesia between May and June 2021. The main data from the nine anchovy purse seine units were collected using purposive sampling. The sampling method was performed by identifying and measuring the components of the number of anchovy purse seiners. The ratio of the length of the head rope to the length of the ground rope is between 0.926 and 0.991, so the net is not exactly rectangular, but almost trapezium in shape. The anchovy purse seine was constructed using webbing polyamide (PA) with 8 mm-9 mm in mesh size, which is considered a small mesh size. It is assembled with some ropes, floats, and sinkers. The anchovy purse seine is characterized by the dominant buoyancy value, 48 % larger than the sinking force. The sinking speed is 0.15 m/s, and the sinking time is 164 s. The anchovy purse seine's design and construction are suitable for operating in the waters of North Coast of Java.

Effect of activated carbon-based two-stage adsorption on biogenic amine reduction and quality of anchovy fish sauce at industrial scale.

This study was aimed to investigate the effectiveness of activated carbon on reduction in biogenic amines (BAs) via two-stage adsorption process at industrial scale, and the consequent effect was evaluated by the taste and aroma of anchovy fish sauce. Through reaction surface methodology, the optimal working paratmeters were determined to adsorbent composition of 2% activated carbon and 0.9% diatomite under temperature of 27 °C for 97 min. Upon optimized settings at industrial scale, there were effective reductions in tryptamine (by 100%), cadaverine (by 10%), histamine (by 61%), and tyramine (by 96%), while the changes in taste-related amino nitrogen, total nitrogen, free amino acids, and color were minimum. In addition, off-flavor-causing compounds, such as alcohols and acids, were removed by the developed method. From the obtained results, the activated carbon-based two-stage adsorption approach can provide the framework for control of BAs contents in fish-based sauces or stocks at commercial and industrial scales.

Efficiency of clustering methods and self-organizing maps of Adriatic sardines and anchovies regarding organochlorine and fatty acid burden.

The Adriatic Sea plays a crucial role as both a significant fishing ground and a thriving trading market for small pelagic edible fish. Recognized for their nutritional value, these fish are esteemed for their high protein content and abundance of polyunsaturated omega-3 and omega-6 fatty acids, making them a sought-after and healthful food choice. Nevertheless, pelagic species can also serve as a reservoir for lipophilic organochlorine pollutants, posing potential risks to human health. In this study, we compared traditional classification methods traditional principal component analysis (PCA) and Ward's clustering with an advanced self-organizing map (SOM) algorithm in determining distribution patterns of 24 organochlorines and 19 fatty acids in sardine and anchovy samples taken from the eastern Adriatic. The outcomes reveal the strengths and weaknesses of the three approaches (PCA, Ward's clustering, and SOM). However, it is evident that SOM has proven to be the most effective in offering detailed information and data visualization. Although sardines and anchovies exhibit similar distribution patterns for p,p'-DDE, PCB-28, PCB-138, PCB-153, PCB-118, and PCB-170, they differ in the concentrations of fatty acids such as stearic, palmitic, myristic, oleic, docosapentaenoic, and docosahexaenoic acid. Our findings supply valuable insights for environmental authorities and fish consumers concerning the potential risks associated with organochlorines in these two types of fish.

SDF-1/CXCR4 signal is involved in the induction of Primordial Germ Cell migration in a model marine fish, Japanese anchovy (Engraulis japonicus).

Primordial germ cells (PGCs) are pivotal for gonadal development and reproductive success. Though artificial induction of sterility by targeting PGCs are gaining popularity due to its advantages in fish surrogacy and biodiversity management, it is often skill and time intensive. In this study, we have focused on understanding the role of PGCs and the chemotactic SDF-1/CXCR4 signaling on gonad development of Japanese anchovy (JA, Engraulis japonicus), an upcoming marine model organism with eco-commercial values, with an aim to develop a novel, easy, and versatile gonad sterilization method. Our data showed that PGC migration related genes, i.e., sdf-1a, sdf-1b, cxcr4a, cxcr4b and vasa, are phylogenetically closer relatives of respective herring (Clupea harengus) and zebrafish (Danio rerio) homolog. Subsequently, PGC marking and live tracing experiments confirmed that PGC migration in JA initiates from 16 hours post fertilization (hpf) followed by PGC settlement in the gonadal ridge at 44 hpf. We found that overexpression of zebrafish sdf-1a mRNA in the germ cell suppresses cxcr4a and increases cxcr4b transcription at 8 hpf, dose dependently disrupts PGC migration at 24-48 hpf, induces PGC death and upregulates sdf-1b at 5 days post hatching. 48 h of immersion treatment with CXCR4 antagonist (AMD3100, Abcam) also accelerated PGC mismigration and pushed the PGC away from gonadal ridge in a dose responsive manner, and further when grown to adulthood caused germ cell less gonad formation in some individuals. Cumulatively, our data, for the first time, suggests that JA PGC migration is largely regulated by SDF1/CXCR4 signaling, and modulation of this signaling has strong potential for sterile, germ cell less gonad preparation at a mass scale. However, further in-depth analysis is pertinent to apply this methodology in marine fish species to successfully catapult Japanese anchovy into a true marine fish model.

Modeling the SDF-1/CXCR4 protein using advanced artificial intelligence and antagonist screening for Japanese anchovy.

SDF-1/CXCR4 chemokine signaling are indispensable for cell migration, especially the Primordial Germ Cell (PGC) migration towards the gonadal ridge during early development. We earlier found that this signaling is largely conserved in the Japanese anchovy (Engraulis japonicus, EJ), and a mere treatment of CXCR4 antagonist, AMD3100, leads to germ cell depletion and thereafter gonad sterilization. However, the effect of AMD3100 was limited. So, in this research, we scouted for CXCR4 antagonist with higher potency by employing advanced artificial intelligence deep learning-based computer simulations. Three potential candidates, AMD3465, WZ811, and LY2510924, were selected and in vivo validation was conducted using Japanese anchovy embryos. We found that seven transmembrane motif of EJ CXCR4a and EJ CXCR4b were extremely similar with human homolog while the CXCR4 chemokine receptor N terminal (PF12109, essential for SDF-1 binding) was missing in EJ CXCR4b. 3D protein analysis and cavity search predicted the cavity in EJ CXCR4a to be five times larger (6,307 Å³) than that in EJ CXCR4b (1,241 Å³). Docking analysis demonstrated lower binding energy of AMD3100 and AMD3465 to EJ CXCR4a (Vina score -9.6) and EJ CXCR4b (Vina score -8.8), respectively. Furthermore, we observed significant PGC mismigration in microinjected AMD3465 treated groups at 10, 100 and 1 × 105 nM concentration in 48 h post fertilized embryos. The other three antagonists showed various degrees of PGC dispersion, but no significant effect compared to their solvent control at tested concentrations was observed. Cumulatively, our results suggests that AMD3645 might be a better candidate for abnormal PGC migration in Japanese anchovy and warrants further investigation.

Development of patty meat analogue using anchovy protein isolate (Stolephorus insularis) as a binding agent.

The development of meat analogues focuses on sustainable production and requires attention to their nutritional, physicochemical, and sensory values. Anchovy protein isolate (API) is a novel and potential binding agent in the development of meat analogues. This study aimed to produce API and evaluate the physical, proximate, and sensory qualities of patty meat analogue (PMA) with the addition of API. The preparation method for API uses pH-shifting. The ratios of API added to the meat analogues were 0 % (F0), 4 % (F1), 8 % (F2), and 12 % (F3) per textured vegetable protein (TVP) weight. Furthermore, PMA was analysed for physical, proximate, and sensory properties. API had 87.23 % dry basis (db) protein content. The amino acid composition of API generally complied with the nutritional requirements of adults and children. The addition of API significantly affected the physical properties, proximate composition, and sensory (taste) qualities of PMA (p < 0.05). The protein content of PMA met Indonesian national standards (SNI) and was similar to both McDonald's and ground beef patty based on United States Department of Agriculture (USDA) standards. F3 was found to be the best based on its physical, proximate, and sensory properties.

Variation in the levels of anisakid infection in the European anchovy Engraulis encrasicolus (Linnaeus) from the Bay of Biscay during the period 2000-2023 (ICES Subarea 8).

The European anchovy Engraulis encrasicolus is one of the most important commercial species in the Bay of Biscay (ICES Subarea 8), and our analysis focused on the analysis of the temporal mean abundance, prevalence, and intensity of Anisakis spp. larvae species in anchovies from ICES Subarea 8 in the years 2000, 2001, 2014-2016, and 2019-2023. Prevalence in adult individuals of anchovy was only 1% in 2000 but increased to 90% in 2014. Since 2015, the prevalence has decreased, and the number of individuals affected in 2023 accounted for 17.6%. The mean abundance showed a similar trend, with a peak of 3.79 nematodes/anchovy in 2014, falling to 0.21 in 2023. The species A. simplex sensu stricto and A. pegreffii were identified by PCR/SANGER sequencing and PCR/RLFP techniques in 2019 and 2020. Anisakis simplex (s.s.) was the most abundant species and, according to the results returned by these two techniques, it accounted for an average of 62.4% and 52.1% of total nematodes in 2019 and 2020, respectively. The results of studies monitoring infection levels in anchovies showed that the mean abundance and prevalence changed over the course of the study period and that the proportion of different species of Anisakis is also subject to variation from year to year.

Shrinking body size of European anchovy in the Bay of Biscay.

Decreased body size is often cited as a major response to ocean warming. Available evidence, however, questions the actual emergence of shrinking trends and the prevalence of temperature-driven changes in size over alternative drivers. In marine fish, changes in food availability or fluctuations in abundance, including those due to size-selective fishing, provide compelling mechanisms to explain changes in body size. Here, based on three decades of scientific survey data (1990-2021), we report a decline in the average body size-length and weight-of anchovy, Engraulis encrasicolus L., in the Bay of Biscay. Shrinking was evident in all age classes, from juveniles to adults. Allometric adjustment indicated slightly more pronounced declines in weight than in total length, which is consistent with a change toward a slender body shape. Trends in adult weight were nonlinear, with rates accelerating to an average decline of up to 25% decade-1 during the last two decades. We found a strong association between higher anchovy abundance and reduced juvenile size. The effect of density dependence was less clear later in life, and temperature became the best predictor of declines in adult size. Theoretical analyses based on a strategic model further suggested that observed patterns are consistent with a simultaneous, opposing effect of rising temperatures on accelerating early growth and decreasing adult size as predicted by the temperature-size rule. Macroecological assessment of ecogeographical-Bergmann's and James'-rules in anchovy size suggested that the observed decline largely exceeds intraspecific variation and might be the result of selection. Limitations inherent in the observational nature of the study recommend caution and a continued assessment and exploration of alternative drivers. Additional evidence of a climate-driven regime shift in the region suggests, however, that shrinking anchovy sizes may signal a long-lasting change in the structure and functioning of the Bay of Biscay ecosystem.

Preliminary data on the occurrence of PAH and Anisakis spp. in Moroccan anchovies: Environmental parasitology and human health risk.

This survey focuses on an environmental parasitology study by investigating Polycyclic Aromatic Hydrocarbons (PAH) bioaccumulation and the occurrence of Anisakis spp. on common anchovies collected from Moroccan coasts over 9 months through chemical (PAH) and Biological (Biometrics and parasitic) analysis. Obtained results were statistically analyzed and human health risks from anchovies consumption were assessed. The results obtained highlighted the good biological and chemical status of this fish in all stations of Morocco's coasts. Anisakis spp. was present in only four stations in the Atlantic fringe (maximum prevalence 22.22 %). Results have shown non-significant differences for tissue nature (Muscle, viscera) or spatial variation and were within a low range of concentrations well below the European Commission standards. Results have shown no serious harmful risk for humans from this fish consumption (ILCR and HI), and statistical analysis had shown positive correlations between prevalence and Chr, prevalence and sex-ratio, and prevalence and weight.

Monthly microplastics change in European anchovy's (Engraulis encrasicolus) gastrointestinal tract in the Black Sea.

To contribute to a better understanding of the regional dynamics of MP pollution and its potential effects on the anchovy population and human health, here we assessed the presence and characterization of microplastics (MPs) in European anchovy (Engraulis encrasicolus), which is the most caught/consumed species in the Black Sea and is of vital importance to the ecosystem. A total of 360 individuals (30 per month) were sampled from the eastern Black Sea continental shelf all year round (monthly from September 20 to Aug 21). We extracted and digested the gastrointestinal tracts (GITs) of the samples with H2O2 and characterized the MPs in the GITs by stereomicroscope and ATR-FTIR. MPs varied between 0 and 0.43 MP individual-1. The morphological structure of MPs was composed of 51 % fiber > fragment (32.7 %) > film (12.2 %) > foam (4.1 %) and polymer types as PP (42.9 %) and PE (22.4 %). In the prevalence of MP colors, black (26.5 %) > white (24.5 %) > red (22.5 %) was observed. The mean MP size was 735.32 ± 836.62 µm, with no significant correlation between the abundance and size of ingested MPs and anchovy height/weight and GIT weight. We determined that MP abundance showed substantial differences between the fishing season (0.18 ± 0.05 MP ind-1) and the banned season (0.05 ± 0.03 MP ind-1). It is essential to develop effective waste management strategies to protect the vulnerable marine ecosystems of the Black Sea and ensure sustainable exploitation of living resources in this region. These strategies should be accompanied by robust monitoring and enforcement measures to guarantee their effectiveness and compliance.

Research and characterization of fibrous microplastics and natural microfibers in pelagic and benthic fish species of commercial interest.

The ingestion of synthetic microfibers, the most prevalent type of microplastics in marine environments, and natural fibers was assessed in Engraulis engrasicolus and Mullus barbatus, two commercially important fish species in the Mediterranean Sea. Microfibers were isolated from the fish gastrointestinal tract using a 10% potassium hydroxide solution. For the microfiber characterization, the evaluation of specific morphological features using a light microscope, coupled with the Fourier-transform infrared (FTIR) analysis of a subsample of isolated particles, was applied. The preliminary results showed the occurrence of microfibers in 53 and 60% of European anchovy and Red mullet, respectively. A mean of 6.9 microfibers/individual was detected in anchovies, while on average Red mullet samples contained 9.2 microfibers/individual. The most common colors of fibers in both species were black, blue, and transparent. Visual characterization of fibers allowed the classification of 40% of the items as synthetic microfibers. FTIR spectroscopy confirmed the visual classification by fiber morphology. Microfibers were made of different typologies of polymers, represented by cellulose, cotton, and polyester. These findings confirm as the wide distribution of fibrous microplastics, and natural microfibers may impact both pelagic and deep-sea trophic webs. Despite the presence of microfibers in fish species poses a potential risk to human health, the literature is scarce regarding studies on the uptake by commercial marine fish mostly due to methodological issues. The visual characterization, corroborated by spectroscopic techniques, may be useful to differentiate synthetic and natural fibers, representing a fast and easy method to assess fibrous microplastic pollution in commercially important fish species.

Incidence of Potentially Toxic Elements and Perfluoroalkyl Substances Present in Canned Anchovies and Their Impact on Food Safety.

Fish plays a key role in a healthy and balanced Italian diet, but it is also subject to the bioaccumulation of different contaminants depending on the geographical or anthropogenic context from which it is derived. In recent years, the European Food Safety Authority (EFSA) has been focusing its attention on consumer toxicological risk, considering emerging contaminants such as perfluoroalkyl substances (PFASs) and potentially toxic elements (PTEs). Regarding fish, anchovies are among the five small pelagic main commercial species in the European Union and the top five fresh species consumed by households in Italy. Considering the lack of data on PFASs and PTEs in this species, our aim was to investigate the mentioned contaminants in salted and canned anchovies collected over 10 months from different fishing areas, even those far apart, to verify possible variations in bioaccumulation and to consider the risk for the consumer. According to our results, the assessed risk was very reassuring also for large consumers. The only concern, related to Ni acute toxicity, also dependent on the different consumers' sensitivity, was related to only one sample.

Preparation of Colorimetric Sensor Array System to Evaluate the Effects of Alginate Edible Coating on Boiled-Dried Anchovy.

The colorimetric sensor array (CSA) is a simple, rapid, and cost-effective system widely used in food science to assess food quality by identifying undesirable volatile organic compounds. As a prospective alternative to conventional techniques such as total volatile basic nitrogen, peroxide value, and thiobarbituric acid reactive substance analysis, the CSA system has garnered significant attention. This study evaluated the quality of edible-coated food products using both conventional and CSA methods in order to demonstrate that the CSA approach is a feasible alternative to conventional methods. Boiled-dried anchovies (BDA) were selected as the model food product, and the sample's quality was assessed as a function of storage temperature and incubation period using conventional techniques and the CSA system. The surface of BDA was coated with an edible alginate film to form the surface-modified food product. The conventional methods revealed that an increase in storage temperature and incubation time accelerated the lipid oxidation process, with the uncoated BDA undergoing lipid oxidation at a faster rate than the coated BDA. Utilizing multivariate statistical analysis, the CSA approach essentially yielded the same results. In addition, the partial least square regression technique revealed a strong correlation between the CSA system and conventional methods, indicating that the CSA system may be a feasible alternative to existing methods for evaluating the quality of food products with surface modifications.

Different Regulatory Effects of Heated Products and Maillard Reaction Products of Half-Fin Anchovy Hydrolysates on Intestinal Antioxidant Defense in Healthy Animals.

The oxidative state of intestinal tracts of healthy animals were investigated after short-term intake of half-fin anchovy hydrolysates (HAHp) and their thermal or Maillard reaction products (MRPs). After one month of continuous oral gavage of HAHp, HAHp-heated products (HAHp-H), the MRPs of HAHp with 3% of glucose (HAHp-3%G MRPs), and the MRPs of HAHp with 3% of fructose (HAHp-3%F MRPs) at a dose of 1.0 g/kg of body weight per day into healthy ICR male mice, the concentrations of serum low-density and high-density lipoprotein cholesterol did not significantly change compared to the control group (CK, gavage with saline). Similar results were found for the interleukin-6 concentrations of all groups. By comparison, HAHp-H, HAHp-3%G MRPs, and HAHp-3%F MRPs administration decreased serum tumor necrosis factor-α concentration as compared to the CK group (p < 0.05). No histological damage was observed in the jejunum, ileum, and colonic tissues of all groups. However, HAHp-H treatment induced higher upregulation of Kelch-like ECH-associated protein 1, transcription factors Nrf-2, associated protective phase-II enzymes of NAD(P)H: quinine oxidoreductase-1, and hemoxygenase-1 in colon tissue, as well as higher upregulation of endogenous antioxidant enzymes, including copper/zinc superoxide dismutase, manganese superoxide dismutase, catalase, and glutathione peroxidase 2 than other groups (p < 0.05). Additionally, increases in Nε-carboxymethyllysine expression in the colonic tissues of all groups were consistent with their increased oligopeptide transporter 1 expressions. Our results suggest that the thermal products of HAHp might have a broad application prospect in improving antioxidant defense in vivo in healthy animals.

Heterologous expression and characterization of Bacillus velezensis SW5 serine protease involved in the hydrolysis of anchovy protein.

BACKGROUND: Bacillus velezensis SW5, with good enzyme production ability, was isolated and identified in our laboratory from fermented fish sauce. Its galactosidase has been expressed in Escherichia coli, which could hydrolyze lactose in milk. The present study aims to express a novel serine protease gene (SPr-SW5) of this strain by Bacillus subtilis WB800N, and applies the expressed enzyme in hydrolysis of anchovy to prepare antioxidant substances, aiming to alleviate the waste of low-value fish resources. RESULTS: SPr-SW5 with the open reading frame of 1353 bp encodes a serine protease (SPr-SW5) with 450 amino acids. The theoretical molecular weight and isoelectric point are 47.2 kDa and 5.22, respectively. The successful expression of SPr-SW5 in B. subtilis WB800N was confirmed by a skim milk plate test. Its optimal temperature and pH were 50 °C and 8.0, respectively. SPr-SW5 activity was increased by Ca2+ and Zn2+ , but inhibited by Fe3+ . Furthermore, SPr-SW5 was tolerant to 1% Tween-40 and Tween-80; however, its activity was strongly inhibited by 10 mm phenylmethylsulfonyl fluoride. Additionally, SPr-SW5 could be capable of hydrolyzing anchovy, the hydrolysate (AHP) at 10 g L-1 , with 2,2-diphenyl-1-picrylhydrazyl and hydroxyl (·OH) scavenging rates of 73.21% and 79.71%, displaying good antioxidant activity. CONCLUSION: The novel SPr-SW5 was successfully expressed in B. subtilis WB800N. It exhibited excellent temperature stability and good tolerance to several metal ions. In addition, the anchovy hydrolyzed by expressed SPr-SW5 has good antioxidant ability. Overall, this research lays a good foundation for SPr-SW5 with respect to exploration and application in the food industry as enzyme preparation. © 2023 Society of Chemical Industry.