Suitability of Antimicrobial Grouper Bone Gelatin Films as Edible Coatings for Vacuum-Packaged Fish Steaks

ABSTRACT

Grouper bone gelatin films were prepared along with chitosan (GC), clove (GL), and pepper (GP) essential oils and examined for their mechanical, barrier, and antimicrobial properties. GC films had a high tensile strength (TS) of 28 MPa, and GL and GP films had a high elongation at break (EAB) of 162 and 166%, respectively. Water solubility and water vapor transmission rate (WVTR) were low in GL films. All the films exhibited inhibitory activities against Staphylococcus aureus, Aeromonas hydrophila, and Listeria monocytogenes. Vacuum-packed fish steaks coated with GC and GL film extended the shelf life from 4 days to 8 days at 4ºC.     

Extraction Optimization and Characterization of Gelatin from Half-Smooth Tongue Sole (Cynoglossus semilaevis Gunther) Skin

Half-smooth tongue sole is a popular warm-water fish, and its skin may be an available resource for gelatin extraction. Response surface method was used to optimize the extraction of gelatin from the fish skin. The most suitable conditions for maximum gelatin yield (24.2%) were pretreatment with 1.9 g/L NaOH, followed by hot water extraction at 63.54ºC for 4.81 h. The obtained gelatin had a high imino acid content (185 residues/1,000 residues), with a high content of crude protein and low levels of ash, fat, and moisture. The predominant sodium dodecyl sulfate–polyacrylamide gel electrophoresis bands revealed that the gelatin contained low-molecular-weight peptides, α-, β-, and γ chains. As compared with bovine gelatin, the fish skin gelatin solution had lower values of strength and clarity and higher levels of L* and viscosity. Additionally, Fourier-transform infrared spectra showed that gel from the gelatin had four amide bands (amide A, amide I, amide II, and amide III) similar to commercial gelatin. The results suggest that half-smooth tongue sole skin could be a promising source of gelatin in view of the comparability between its gelatin and commercial gelatin in physicochemical properties.     

北极微生物的分离及抗菌抗肿瘤活性筛选

采用多种培养基对北极地区(海洋沉积物、湖泊沉积物及土壤)微生物进行分离,并采用琼脂平板打孔法和MTT法进行了抗菌、抗肿瘤活性菌株的筛选.共分离极地微生物151株,筛选结果显示,12.4%的菌株有抗菌活性,6.2%菌株有抗肿瘤活性,其中3株同时具有抗菌、抗肿瘤活性,表明极地有良好的微生物药用资源.     

马氏珠母贝鳃组织抗菌活性的研究

用纸片琼脂扩散法和微量测定法对制备的马氏珠母贝鳃组织匀浆物进行抗菌活性分析.结果表明,马氏珠母贝鳃组织匀浆物具有抗革兰氏阴性菌-副溶血弧菌、河流弧菌、嗜水气单胞菌、大肠杆菌、绿脓杆菌活性;对革兰氏阳性菌-金黄葡萄球菌具一定抑制活性,但对枯草芽孢杆菌无抑制活性.利用胰蛋白酶对马氏珠母贝鳃组织匀浆物水解后,其抗菌活性消失,表明其中抗菌成分为蛋白质.将马氏珠母贝鳃组织匀浆物在不同温度(37～95 ℃)和不同pH(2.5～5)保温,发现其抗菌活性成分对热及酸耐受性较强.马氏珠母贝鳃组织匀浆物在低浓度无溶血活性.该抗菌组分在马氏珠母贝的免疫防御中具有一定的作用.     

Assessment of the Antibacterial Effect of Chitosan Coated with Heracleum persicum Oil on Rainbow Trout Quality

Today, due to the rapid spoilage of fish, the use of natural preservatives is a priority over those of synthetic varieties. Also, the natural antimicrobial effects of essential oils can help to increase shelf life. There are very few studies concerning the use of essential oils in this regard. In the present study, the antibacterial effect of chitosan coated with Heracleum persicum oil was investigated on the quality of rainbow trout. The control and the coated fish samples were analyzed periodically by generalized estimating equation (GEE) for total viable count (TVC), psychrophilic bacteria, total volatile base nitrogen (TVB-N), polyunsaturated fatty acids (PUFA), texture, taste, odor, and overall acceptance parameters. The results showed that by increasing the amount of chitosan, TVC, psychrophilic bacteria, and TVB-N decreased while PUFA, texture, taste, odor, and overall acceptance parameters increased.     

盐酸恩诺沙星与其他四种抗菌药物对水产致病菌的体外抗菌作用

用琼脂平板稀释法测定了盐酸恩诺沙星、盐酸环丙沙星、氟哌酸、米诺沙星和氟苯尼考对常见水产致病菌的最低抑菌浓度(MIC)和最低杀菌浓度(MBC)，结果表明：同一抗菌药物对不同致病菌的抗菌效果存在较大差异，不同抗菌药物对同一致病菌的抗菌效果也存在较大差异。使用4倍MBC的药物浓度对嗜水气单胞菌、温和气单胞菌和非O1-群霍乱弧菌杀菌时间随药物种类和致病菌不同而存在差异。     

The Effects of Grass Carp Skin Gelatin and Whey Protein Interactions on Rheological and Textural Properties and Nanostructure

The physical properties and nanostructure of fish skin gelatin with and without whey protein isolate (WPI) were investigated. The addition of low concentrations of WPI (1 and 3%, w/v) into fish gelatin decreased the gel strength and textural properties. The addition of 5% WPI did not significantly influence textural properties but increased the melting and gelling temperatures of fish skin gelatin and improved the appearance of gelatin gels. X-ray diffraction indicated that a higher degree of crystallinity with an ordered structure or ordered structure snippets occurred with increasing WPI concentrations in line with the slight decrease in the loss factor (tan δ) with time sweeps. Fish gelatin had a spherical nanostructure, and a homogeneous nanostructure was observed in gelatin with 3% WPI. The addition of 5% WPI led to large aggregates in gelatin gels. The results indicated that whey protein could affect the nanostructure of fish skin gelatin gels and thus change their gelation properties and texture in food and pharmaceutical applications.     

Characteristics and Physicochemical Properties of Gelatin Extracted from Scales of Seabass (Lates calcarifer) and Grey Mullet (Mugil cephalus) in Vietnam

In this study, gelatin was obtained from the scales of seabass (Lates calcarifer) and grey mullet (Mugil cephalus) collected in Vietnam by using Alcalase® 2.4 L FG for pretreatment and electrochemically activated water with pH 2.0–2.5 for extraction. The characteristics and physicochemical properties of gelatin were investigated. The results indicated that the gelatin had a high protein content (88.6–90.0%) with ash (1.43–1.55%) and no fat. The gelatin was identified as type A due to its pH value. The yields of gelatin were 14.1–15.2%. The gel strength and melting point were 270.3 g and 26.7°C, respectively, for gelatin from seabass scales and 249.1 g and 25.9°C, respectively, for samples from grey mullet scales. In contrast, the viscosities of gelatin were found to be 6.97 cP for seabass and 8.73 cP for grey mullet. Both gelatins contained α-chain and β-chain as the major components. Circular dichroism spectra indicated that the triple helical structure in collagen was almost transformed to the random coil structure in gelatin. The proportion of imino acids was high (197–205 residues/1,000 residues). So, gelatin from seabass and grey mullet scales could be used as a potential replacement for mammalian gelatin.     

Characterization and Functional Properties of Gelatin Extracted from Chinese Giant Salamander (Andrias Davidianus) Skin

The characteristics and functional properties of gelatin obtained from Chinese giant salamander (Andrias davidianus) skin were investigated. After alkaline and acetic acid pre-treatment, yield of the gelatin by hot water extraction (60°, 8 h) was 17.63 g/100 g (wet basis), and the gelatin contained 89.46 g/100 g protein, 0.3 g/100 g fat, and 0.41 g/100 g ash (dry basis). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results of the gelatin showed clear band pattern for collagen subunits,β- and α-chains. The gelatin contained 18.85% imino acids, 28.62% hydrophobic amino acids, and 19.68% essential amino acids, which is slightly lower than porcine skin gelatin (PSG). The gelatin exhibited thermo-reversible properties, with gel strength of 192.50 ± 11.47 g, rupture distance of 10.17 ± 0.20 mm, gelling temperature of 4.98°, and melting temperature of 18.04°, Emulsifying activity index (EAI) of the gelatin diminished with increasing concentrations from 10 to 40 mg/mL (p < .05), whereas the emulsifying stability index (ESI) improved significantly (p < .05). Higher concentrations (30 and 40 mg/mL) could significantly enhance the gelatin’s foam expansion (FE) and foam stability (FS) (p < .05). Water absorption ability and oil absorption ability of the gelatin were 7.1 g water/g gelatin and 9.8 g oil/g gelatin, respectively. These results imply that the gelatin with good interfacial properties could potentially be used as a novel ingredient in food emulsion systems.

Abbreviations: CGSS, Chinese giant salamander skin; CSG, Chinese giant salamander skin gelatin; PSG, porcine skin gelatin; EAI, emulsion activity index; ESI, emulsion stability index; FE, foam expansion; FS, foam stability; WAC, water absorption capacity; OAC, oil absorption capacity.     

大弹涂鱼皮肤黏液的抑菌活性和蛋白质组学

皮肤黏液是鱼类免疫防御的第一道防线。大弹涂鱼皮肤黏液对其免疫防御、渗透压维持以及适应水陆生活等方面具有重要意义。为深入了解大弹涂鱼皮肤黏液的蛋白质分子组成,对其皮肤黏液开展了抑菌活性分析和蛋白质组学分析。通过电刺激法收集大弹涂鱼皮肤黏液,采用打孔法比较了皮肤黏液和血清的抑菌活性差异;进一步利用鳗弧菌对大弹涂鱼进行诱导,采用生长曲线抑制法分析和比较了诱导前后皮肤黏液的抑菌活性差异。利用Shotgun质谱技术,结合大弹涂鱼皮肤转录组数据库,对大弹涂鱼皮肤黏液开展了蛋白质组学分析;进一步利用String软件对所鉴定的蛋白质开展了蛋白质相互作用预测。大弹涂鱼皮肤黏液具有广谱抑菌活性,鳗弧菌诱导后的大弹涂鱼皮肤黏液与诱导前相比,对部分革兰氏阴性菌的抑菌活性明显上升,但对其他菌株的抑菌活性差异不明显。从大弹涂鱼皮肤黏液中共鉴定各类蛋白质分子97种,基本分子组成与其他硬骨鱼类皮肤黏液蛋白相似,但也有少数蛋白如泛素蛋白、胸腺素蛋白等未在其他鱼类皮肤黏液中报道。此外,其他鱼类中所鉴定到的部分蛋白如热休克蛋白、抗菌肽等在大弹涂鱼皮肤黏液中未能鉴定到。大弹涂鱼皮肤黏液中已鉴定的蛋白多数具有与免疫功能相关的结构域,且存在一个以肌动蛋白为核心的相互作用网络。大弹涂鱼皮肤黏液具有明显的广谱抑菌活性,其蛋白质组成与其他鱼类皮肤黏液具有一定的相似性。本研究为深入了解大弹涂鱼皮肤黏液的分子多样性及功能机制奠定了基础。     

Development of Optimized Edible Packaging Based on White-cheek Shark (Carcharhinus dussumieri) Skin Gelatin Biopolymer: Mechanical,Water Vapor Permeability,and Structural Properties

White-cheek shark skin gelatin (WSG) edible films were optimized with respect to their barrier, mechanical, and structural properties using response surface methodology (RSM), with the introduction of gelatin concentrations (1–4% w/v based on film forming solution) and glycerol contents (15–35% w/w based on gelatin weight) as the independent variables. WSG films with moderate levels of glycerol had higher tensile strength (TS), elastic modulus (EM), and elongation at break (E@b) depending more significantly on gelatin content. Adding higher levels of glycerol, film roughness was observed to decrease exponentially but water vapor permeability (WVP) increased linearly. Structural analysis using Fourier transform infrared spectroscopy (FTIR) showed more aggregated protein structures at higher gelatin content, which could be strongly related to the notable increase of contact angle. According to the model, the optimum conditions for the WSG film formula were found to be 3.3% WSG and 19.3% glycerol, giving a predicted maximum E@b, TS, EM, roughness, and contact angle of 183%, 4.4 MPa, 92 MPa, 4.93 nm, and 101°, respectively, with a predicted minimum WVP of 11.8 g.mm/day.m².kPa.     

Rheological,Physico-chemical,and Surface-Active Properties of Gelatin Extracted from Bigeye Tuna (Thunnus obesus) Skin Waste

ABSTRACT

In the present investigation, bigeye tuna (Thunnus obesus) skin waste was utilized for the extraction of gelatin. Physico-chemical, rheological, and surface-active properties of the obtained gelatin were assessed. Proximate compositional analysis of tuna skin gelatin recorded high protein content of 88.6% and low levels of fat and ash contents. The isoelectric point (IEP) of tuna skin gelatin was around 6.24. Amino acid profiling recorded a glycine [31.7%] higher proportion, followed by imino acids (hydroxyproline + proline) [18.4%], and alanine [12.2%]. The electrophoretic pattern of purified gelatin exhibited three major bands (β, α1, and α2) with wider molecular weight distribution in the range of 200–120 kDa. The Fourier Transform Infrared (FT-IR) spectra of gelatin also confirmed the bands corresponding to α helix, β sheet, and random coil structures. The rheological studies of tuna skin gelatin revealed Newtonian and shear thickening properties. The method of extraction and assessment of its characteristic properties could be useful in the field of formulation of functional foods, nutraceutical, and biomedical applications.     

Anti-Inflammatory,Antioxidant, and Antimicrobial Effects of Underutilized Fish Protein Hydrolysate

Antimicrobial, anti-inflammatory, and antioxidant activities of protein hydrolysates from Argentine croaker (Umbrina canosai) protein isolate (CPI) or Argentine croaker myofibrillar protein with different degrees of hydrolysis (DH: 10–20%) prepared using Alcalase or Protamex were determined. Results showed that an increase in the DH resulted in higher content of hydrophobic and aromatic amino acids (AAAs) and in a decrease in molecular weight (MW) distribution for all hydrolysates obtained. Furthermore, the enzyme and raw material used influenced the amino acid content and MW determined. Hydrolysates from CPI with a 20% DH by Alcalase had higher 2,2?-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activity, metal chelation, and ferric-reducing antioxidant power (p < 0.05). All hydrolysate samples decreased the pro-inflammatory capacity. In all the evaluated microorganisms, only seven were inhibited, most being Gram-positive. Alcalase was found to exert a considerable influence on antibacterial activity. These hydrolysates are an alternative as natural antimicrobials, anti-inflammatory, and antioxidant compounds.     

In vitro assessment of the antimicrobial efficacy of chitosan nanoparticles against major fish pathogens and their cytotoxicity to fish cell lines

Nanotechnology is an emerging avenue employed in disease prevention and treatment. This study evaluated the antimicrobial efficacy of chitosan nanoparticles (CSNPs) against major bacterial and oomycete fish pathogens in comparison with chitosan suspension. Initially, the minimum inhibitory concentrations (MIC, MIC₉₀) were determined and the per cent inhibition of bacterial growth was calculated. Subsequently, the minimum bactericidal concentrations (MBCs) were determined. The time-dependent disruptions of CSNP-treated pathogens were observed via transmission electron microscopy (TEM), and the effect of CSNPs on the viability of two fish cell lines was assessed. No antimicrobial effect was observed with chitosan, while CSNPs (105 nm) exhibited a dose-dependent and species-specific antimicrobial properties. They were bactericidal against seven bacterial isolates recording MBC values from 1 to 7 mg/ml, bacteriostatic against four further isolates recording MIC values from 0.125 to 5 mg/ml and fungistatic against oomycetes recording MIC₉₀ values of 3 and 4 mg/ml. TEM micrographs showed the attachment of CSNPs to the pathogenic cell membranes disrupting their integrity. No significant cytotoxicity was observed using 1 mg/ml CSNPs, while low dose-dependent cytotoxicity was elicited by the higher doses. Therefore, it is anticipated that CSNPs are able to compete and reduce using antibiotics in aquaculture.     

The protective effects of Vitamin C and Trolox on kinematic and oxidative stress indices in rainbow trout sperm cells against flower‐like ZnO nanoparticles

This study investigated the protective effects of 1 mM Vitamin C and 1 mM Trolox on kinematic and oxidative stresses in rainbow trout (Oncorhynchus mykiss) sperm cells against flower‐like zinc oxide nanoparticles (ZnO‐NP, 1 mg/L) in vitro, during 2 hr. Trolox showed protective effects on all kinematic parameters measured for sperm cells against ZnO‐NPs, whereas Vitamin C only affected angular path velocity (VAP), linearity (LIN) and amplitude of lateral displacement of the sperm cell head (ALH). Although ZnO‐NPs reduced total glutathione (TGSH) and catalase (CAT) compared with the control, there were positive treatment effects for both Trolox and Vitamin C on antioxidant capacity. Conversely, while ZnO‐NP increased the rate of malondialdehyde (MDA) lipid peroxidation, it decreased following antioxidant therapy. This research suggests that Trolox and Vitamin C play a protective role for rainbow trout sperm exposed to ZnO‐NP.     

Antioxidant,ACE-Inhibitory,and Antimicrobial Activities of Peptide Fractions Obtained From Dried Giant Squid Tunics

ABSTRACT

Squid tunics were divided in two batches: freeze-dried and air-dried. Dried squid tunics were directly hydrolyzed with pepsin, Alcalase, and Esperase. Freeze-dried tunics showed better aptitude for hydrolysis than air-dried tunics. Pepsin showed the lowest efficiency in protein breakdown and gave hydrolysates with low antioxidant activity, whereas Alcalase and Esperase peptide fractions showed strong radical scavenging ability, ACE-inhibitory activity, and noticeable antimicrobial properties. B. cereus, B. coagulans, and D. hansenii were found to be the most sensitive bacteria. Dried squid tunics proved to be an alternative to a previously extracted gelatin for obtaining bioactive peptides.     

Characterization of Spray-Dried Hydrolyzed Proteins from Pink Perch Meat Added with Maltodextrin and Gum Arabic

In the present study, spray-dried pink perch meat protein hydrolysate (PPH) and hydrolysate with maltodextrin and gum arabic (PPHMG) were characterized in relation to their physical, rheological, functional, antioxidant, thermal, and sensory properties. The Herschel–Bulkley model was suitable to explain the flow behavior of PPH and PPHMG emulsions. Both the samples exhibited functional and antioxidant properties. Antioxidant activities were significantly higher for PPH compared with PPHMG (p<0.05). PPH and PPHMG alone did not exhibit antimicrobial activity but enhanced the activity of chitosan against pathogens. Sensory acceptability of the samples in soup revealed that PPHMG of up to 4% was highly scored without any traces of bitterness, whereas for PPH, a concentration of up to 3% was acceptable.     

Optimized Extraction,Preliminary Characterization and Evaluation of the in Vitro Anticancer Activity of Phlorotannin-Rich Fraction from the Brown Seaweed,Cystoseira sedoides

Brown seaweeds produce useful bioactive substances with high cosmetic and pharmacological values due to the presence of antioxidant derivatives, mainly phlorotannins (PHT), which are of particular interest. A central composite rotatable design (CCRD) with three variables (extraction time, dry material-to-solvent ratio and ethanol concentration) and two responses was performed to optimize the microwave-assisted extraction (MAE) of phlorotannins from Cystoseira sedoides using response surface methodology (RSM). Results showed that the independent variables significantly affected both phlorotannin content and the scavenging capacity. The optimum operating conditions were extraction time, 101.74 sec; dry material-to-solvent ratio, 1:10 g/mL; and ethanol extraction, 50%. Under these conditions, the predicted values of PHT content and radical scavenging activity-IC50 were close to the observed values and were 383.887 µg PGE/g Dm and 18.353 µg/mL, respectively. Characterization of the phlorotannin-rich fraction was conducted by high-performance liquid chromatography and Fourier transform infrared spectroscopy. The evaluation of the anticancer activity against MCF-7 breast cancer cell line showed a potent activity to trigger apoptotic death in more than a half of the MCF-7 cells, with an estimated IC50 value of 78 μg/mL. In addition, this fraction induced a notable growth regression effect on 3D spheroids model in a concentration-dependent manner, with a growth rate of about 1.17, at 200 µg/mL.

Abbreviations: CCRD: Central composite rotatable design; Dm: Dry material; DMBA: 2,4-dimethoxy benzaldehyde; DMSO: Dimethyl sulfoxide; DPPH: 1,1-diphenyl-2-picrylhydrazyl; EDTA: Ethylene diamine tetraacetic acid; FBS: Fetal bovine serum; FT-IR: Fourier transform infrared; HPLC: High-performance liquid chromatography; IC₅₀: Half-maximal inhibitory concentration; MAE: Microwave-assisted extraction; Min: minutes; PBS: Phosphate buffered saline; PGE: Phloroglucinol equivalent; PHT: Phlorotannins; PHT-SED: phlorotannins derived from Cystoseira sedoides; PI: Propidium iodide; RSA: Radical scavenging activity; RSM: Response surface methodology; Sec: seconds     

Effect of oral administration of l -thyroxine (T4) on growth performance, digestibility, and nutrient retention in Channa punctatus (Bloch) and Heteropneustes fossilis (Bloch)

Effects of oral administration of l-thyroxine (T₄) on growth performance, body composition, and some aspects of nutritional physiology were investigated in two slow-growing air-breathing fish (Channa punctatus and Heteropneustes fossilis) under laboratory conditions (LD 12:12 at 25°C). The results indicate that irrespective of the species, fish fed diets containing lower doses of T₄ (up to 50 mg kg⁻¹ of diet in C. punctatus and up to 100 mg kg⁻¹ of diet in H. fossilis) showed significantly (P < 0.05) higher growth (in terms of live weight and length gain, specific growth rate, percentage gain in body weight and condition factor), low feed conversion ratio, high nutrient retention, high apparent protein digestibility, and high digestive enzyme activity. Viscero-somatic (VSI) and hepato-somatic (HSI) values were also high in fish fed on low dietary T₄ levels. Liver glycogen levels decreased with the increase in the dietary T₄ levels, indicating its utilization during growth. Muscle glycogen levels in H. fossilis coincided with high growth at 100 mg of T₄. Observation of the postprandial excretion of metabolites (N–NH₄ ⁺ and o-PO₄ ⁻) indicated significantly (P < 0.05) low levels in aquaria water where the fish were fed diets with low T₄ levels. These studies further revealed that feeding fishes on higher T₄ levels (above 50 mg in C. punctatus and above 100 mg in H. fossilis) not only repressed growth performance and nutrient retention, but also affected carcass composition by lowering protein accumulation (muscle and carcass protein) and energy assimilation. These studies revealed a biphasic action of thyroxine, i.e., the hormone at lower doses is anabolic, while at higher doses it acts as a catabolic agent, indicating that feeding fishes on higher doses can be detrimental to their growth and metabolism. In summary, the results of the present study show that feeding H. fossilis and C. punctatus on low doses of T₄ enhances growth, decreases excretion of metabolites, and increases nitrogen retention. These observations suggest that T₄ supplementation of the diet may have practical utility in the culture of slow-growing fish species.     

循环禁食对黄鳝血清溶菌酶活力和替代补体溶血活性的影响

研究循环性禁食后黄鳝(Monopterus albus)非特异性免疫力的变化,为生产实践中制定可行的禁食投喂方案提供更好的依据。试验用幼鳝体重(19.5±2.0)g,饲养在实验室的塑料水族箱(70 cm×55 cm×36 cm)中,正式试验前进行1周饱食投喂适应性暂养,暂养和试验期间的饵料为鲜活的水蚯蚓。循环禁食处理分2种:一种为不同比例的禁食时间与投喂时间的循环性禁食处理,即不同水平的循环性禁食处理;另一种为禁食时间与投喂时间的比例一致但禁食与投喂的时间间隔不同的循环性禁食,即不同时间间隔循环性禁食处理。结果表明,不同水平循环禁食试验组黄鳝其血清溶菌酶活力均显著低于对照组(P0.05),试验组之间无显著差异(P0.05);血清替代补体溶血活性显著高于对照(P0.05),试验组之间无显著差异(P0.05)。不同时间间隔循环禁食试验组黄鳝血清溶菌酶活力均显著低于对照组(P0.05),随着禁食持续时间延长溶菌酶活力呈逐渐下降趋势;血清替代补体溶血活性均显著高于对照组(P0.05),试验组之间无显著差异(P0.05)。采取特定投喂模式促使鱼类获得生长补偿效应的同时其非特异性免疫力不受影响是有可能实现的。