Essay of Different Extraction Procedures in Capelin Fish Meal for Biogenic Amine Determination by HPLC

This study assesses the effects of different extraction procedures of biogenic amines in capelin fish meal with different concentrations of trichloroacetic acid (TCA) for their determination by high performance liquid chromatography (HPLC). No significant differences were noted between the use of TCA at 7.5 and 10% for the extraction of biogenic amines in E1 and E3 extraction, except for tyramine. The extraction using 7.5% TCA in one step and centrifugation (E1) was a simple and rapid method. This extraction seems to be the most suitable for the extraction of amines in these kinds of samples, compared to the other methods tested. The biogenic amine that presented the highest levels in the fish meal studied was cadaverine (208–226 mg/kg), followed by putrescine and histamine (164–173 and 44–46 mg/kg, respectively). Biogenic amine levels were low when compared to the toxic levels observed in these kinds of products.     

Comparison of Lactic Acid Bacteria Fermentation with Acid Treatments for Chitosan Production from Shrimp Waste

The traditional procedure for chitosan production involves use of a strong acid (HCl) for demineralization of chitin. This study reports application of a mixed culture of lactic acid bacteria (Lactobacillus plantarum, Lactobacillus acidophilus, and Lactobacillus lactis) fermentation in demineralization of chitin for chitosan production from shrimp waste. Chitosan produced from shrimp waste with lactic acid bacteria fermentation at 30°C for 72 h or 1 M lactic acid treatment at room temperature for 2 h followed by alkaline treatments (0.5 M NaOH at 25°C for 4 h followed by 12.5 M NaOH at 70°C for 10 h) contained very low protein (0.8–1.1%) and ash (<0.01%) contents with the same solubility (100%) as those of chitosan produced with hydrochloric acid treatment. Lactic acid bacteria fermentation and lactic acid treatment may be utilized in chitosan production from shrimp waste to reduce use of strong chemicals.     

Application of Native Lactic Acid Bacteria (LAB) for Fermentative Recovery of Lipids and Proteins from Fish Processing Wastes: Bioactivities of Fermentation Products

Fermentation using native lactic acid bacteria (LAB) was evaluated for its effectiveness in recovering lipids and proteins simultaneously from freshwater fish visceral waste (FVW). Five different LAB isolated from fish processing waste were employed in a fermentation process that involved 10% (w/w) glucose, 2% (w/w) NaCl, and 10% (v/w) LAB. Cultures evaluated included four native isolates (Pediococcus acidilactici NCIM5368, Enterococcus faecalis NCIM5367, Pediococcus acidilactici FM37, and Pediococcus acidilactici MW2) from FVW with E. faecium NCIM5335 as the reference culture. Fermentation with native LAB resulted in recovery of > 90% oil present in the material as against no recovery in case of raw (unfermented) viscera and resulted in > 50% of degree of hydrolysis of proteins. The fatty acid profile of lipids was not affected by the fermentation process. The fermentation liquor, rich in hydrolyzed protein, exhibited antioxidant as well as antagonistic properties against several bacterial pathogens. The results clearly demonstrate the usefulness of fermentation using native isolates for simultaneous recovery of lipids and proteins from fish processing waste. It also asserts the value of fermentation as an eco-friendly method and aids in minimizing disposal/pollution problems associated with these solid wastes.     

Isolation,Screening, and Identification of Proteolytic Lactic Acid Bacteria from Indigenous Chao Product

The aim of the study was to isolate, select, and identify proteolytic lactic acid bacteria (LAB) from chao, a traditional fermented fish from Pangkajene and Kepulauan Regency, South Sulawesi, Indonesia. LAB was isolated by poured plate method. Proteolytic LAB were selected using agar skim milk media. Protease activity of LAB was determined based on the amount of tyrosine released in unit/mL. Proteolytic LAB were identified using API 50 CHL kit and 16S rRNA gene sequence analysis. The result showed that a total of 60 isolates were obtained from chao, and 57% of them were cocci-shape. Fifteen isolates were halotolerant proteolytic LAB. Their R values and protease activity were 2.11–3.39 and 0.267–0.304 U/mL, respectively. Identification by API 50 CHL kit showed that four rod-shape isolates were Lactobacillus plantarum, and two others were Lactobacillus curvatus. Cocci-shape isolates could not be identified as cocci bacterium. Rep-PCR results showed that there were two kinds of bands, namely thick and thin. Two isolates were selected from two types of bands that had the highest R for the analysis of 16S rRNA gene sequences, namely Ags1-3 and Ags7-3. The results showed that Ags1-3 isolate was identified as Lactobacillus plantarum and Ags7-3 as Pediococcus acidilactici.     

Effects of Lactic Acid Bacteria and Citrus Essential Oil on the Quality of Vacuum-Packed Sea Bass (Dicentrarchus labrax) Fillets During Refrigerated Storage

The preservative effect of refrigerated and vacuum-packed sea bass (Dicentrarchus labrax) fillets inoculated with four mixtures of lactic acid bacteria (LAB) (Lactococcus lactis, Lactobacillus plantarum, and Carnobacterium piscicola) and incorporated with citrus essential oil (CEO) was evaluated on the basis of microbiological and biochemical analysis.

Initially, sea bass fillets showed high nutritional quality. During refrigerated storage, lipid contents did not show a significant decrease in any fish fillets; meanwhile, important proteolysis was observed in untreated control. In addition, results indicated that both CEO and LAB strains exhibited antimicrobial activity against spoilage, pathogenic, and fungi flora. Moreover, the total volatile bases (TVB-N) values were higher in control fillets, and the lowest TVB-N values were observed in Control CEO and C3 + CEO (30.47 ± 0.00 and 32.29 ± 1.12 mg TVB-N/100 g, respectively). Also, the levels of biogenic amines increased in all fillets without exceeding the upper limit of acceptability except for untreated control (sum of about 1396.63 ppm). Furthermore, this combined treatment ameliorated the muscle liquid-holding capacity, which improves technological properties.

Overall, this treatment may open new promising opportunities for the biopreservation of fish products by enhancing the period of storage of refrigerated and vacuum-packed sea bass fillets.     

Effect of Bacterial Community and Free Amino Acids on the Content of Biogenic Amines During Fermentation of Yu-lu,a Chinese Fermented Fish Sauce

The main objective of this work was to investigate the effects of bacterial community and free amino acids on the content of biogenic amines in Yu-lu during fermentation. Four major biogenic amines (histamine, tyramine, cadaverine, and putrescine) were identified, and the content of tyramine was positively correlated with histamine (R value = 0.9113). Aspartic acid, glutamic acid, histidine, leucine, and lysine were the dominant free amino acids. 16S rRNA sequencing showed that the composition of the bacterial community changed significantly during fermentation of Yu-lu. Principal component analysis revealed the crucial links between microbial community and biogenic amines. For example, Halanaerobium was probably associated with the formation of putrescine, while Halomonas might be associated with the degradation of biogenic amines at the end of fermentation of Yu-lu. This study provided a detailed evaluation of the Yu-lu fermentation process, enabling development of better strategies for biogenic amine control in fish sauce.     

鱼类细菌性败血症发病鱼池细菌数量及致病性气单胞菌的初步研究

本文对细菌性败血症发病鱼池细菌数量及气单胞菌的数量、溶血性及致病力进行了研究。结果表明:水温在28.5℃以下时,细菌数量与水温基本成正比,发病池与不发病池的细菌数量无显著性差异;水体中广泛存在气单胞菌,其占水体总菌量比例随水温的上升而升高;发病池及未发病池中均存在能产生hec毒素和不产生hec毒素的嗜水气单胞菌株,两者数量上无显著差异;水体中存在血清型为TPS—30及PBJS—76的嗜水气单胞菌,且大多数对鱼体有毒力,以5×10~7CFU/尾腹腔注射,可使试验鱼大部分或全部发病死亡。     

Isolation,Identification and Characterization of Lactic Acid Bacteria From Shao-jiou-luo (Fermented Zoned Cerith), a Traditional Fermented Food in Taiwan

This study investigated the distribution of the native lactic acid bacteria (LAB) population in shao-jiou-luo (fermented zoned cerith), a popular traditional fermented food in Taiwan. A total of 129 cultures of LAB were isolated from shao-jiou-luo samples, and 14 cultures were isolated from its main raw material, zoned cerith. These isolates were divided into classes by phenotype and then into groups by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. The identification results revealed 11 bacterial species belonging to five genera in shao-jiou-luo and three species belonging to two genera in zoned cerith. The most common bacterial genera in shao-jiou-luo were heterofermentative Weissella and Leuconostoc. On the other hand, Enterococcus aquimarinus was the most common species found in the raw zoned cerith sample. In conclusion, the results suggest that heterofermentative LAB species greatly contributed to the fermentation of shao-jiou-luo. This is the first report describing the distribution and varieties of LAB that exist in the shao-jiou-luo fermentation process and raw zoned cerith.     

The Effects of Chitosan Coating on Biogenic Amines Inhibition and Microbial Succession of Refrigerated Grass Carp (Ctenopharyngodon idellus) Fillets

Chitosan coating and plastic wrap coating were investigated for their effects on the quality of grass carp fillets stored at 4 ± 1ºC for a period of 20 days. The fillets were divided into groups of untreated (Control), wrapped with sterile preservative film (PE), and chitosan-treated (1, 2, and 2.5% w/v corresponding to CH1, CH2, and CH2.5, respectively). The final result demonstrated that chitosan coating has positive effects on the microbial inhibition of total volatile count (TVC), lactic acid bacteria (LAB), Enterobacteriaceae, Pseudomonas, H₂S-producing bacteria (including Shewanella putrefaciens), and Staphylococcus. With regard to the results of biogenic amines (BAs), putrescine (PUT) and histamine (HIS) were the major amines, and their concentration was significantly (p < 0.05) higher in Control and PE than in chitosan-coated fillets at the end of storage. The final concentration in Control, PE, CH1, CH2, and CH2.5 was 143.7, 160.1, 83.0, 59.4, and 44.2 mg/kg for PUT and 228.0, 57.1, 4.1 3.2, and 2.9 for HIS, respectively. Chitosan coating was found to inhibit the increase of total volatile base nitrogen (TVB-N), microbial population, and total BAs and prolong the shelf life of grass carp fillets. However, sterile plastic wrap coating had little positive effect on maintaining the quality of grass carp fillets during refrigerated storage.     

The Effect of K-Lactate Salt and Liquid Smoke on Bacterial Growth in a Model System

The effects of a commercial lactate salt formulation—containing potassium lactate (KL) and potassium acetate (KA)—and liquid smoke (LS) on the growth of selected lactic acid bacteria (LAB; Carnobacterium maltaromaticum, Carnobacterium inhibens, Lactococcus lactis, Lactobacillus curvatus, and Enterococcus faecalis), fish spoilage bacteria (Photobacterium phosphoreum, Pseudomonas putida, and Vibrio vulnificus), and Listeria innocua, were examined in a tryptic soy broth model system at 20ºC based on BioScreen data. The most pronounced inhibition effect on growth of bacteria was seen in the presence of 6% KL + 0.4% KA, used either combined with LS or alone. Only a minor inhibition effect on growth was found in the presence of LS alone. The only exception was Lactobacillus curvatus, which grew quite well in the presence of LS compared to control medium. The growth of Vibrio vulnificus was prevented in 6% KL + 0.4% KA, and significantly inhibited in the presence of 3% KL + 0.2% KA. When V. vulnificus was grown in NaCl, KA, and KL + KA, it was observed that KL + KA had a better inhibition effect than sodium salt within the same concentration range.     

脂肪酸营养对鱼类生长、脂代谢及免疫性能影响的研究进展

脂肪酸在鱼类营养中起着重要的作用,脂肪酸作为能源物质为鱼类的生长发育等各项活动提供能量,并协助脂溶性维生素的吸收和体内运输。不同鱼类因其生活环境不同,对必需脂肪酸的需求种类也不同。由于不同脂肪水平与不同脂肪酸来源的饲料因其脂肪酸含量或脂肪酸组成不同,不同鱼类或鱼类的不同生长阶段对脂肪源的利用也不同,那么饲料的脂肪水平或脂肪酸组成不同就对鱼类的生长、脂代谢及免疫性能产生一定的影响作用。必需脂肪酸作为鱼类重要的脂肪酸营养物质同样影响鱼类的生长、脂代谢及免疫性能。本文从脂肪酸营养对鱼类生长、脂代谢及免疫性能的影响等方面进行综述,并对今后的研究重点进行了展望。’     

The Effects of Liposomal Encapsulated Thyme Extract on the Quality of Fish Mince and Escherichia coli O157:H7 Inhibition During Refrigerated Storage

The effects of liposomal encapsulated and unencapsulated (0.3 and 0.5% w/w) thyme extract (TE) on the quality of silver carp (Hypophthalmichthys moltrix) mince during chilled storage (4 ± 1°C) were examined over a period of 15 days. The samples were analyzed by microbiological (total viable count, psychrotrophic count) and chemical (peroxide value and total volatile nitrogen) experiments. Also, the efficacy of the encapsulated extract in control in the population of Eschershia coli O157:H7 inoculated in silver carp mince was investigated. Results showed that the samples containing 0.5% of encapsulated thyme had fewer changes in peroxide value and total volatile nitrogen in comparison to other treatments (p < 0.05). Encapsulated thyme also could inhibit the growth of mesophilic and psychrotrophic bacteria in the samples compared to the control and the samples containing unencapsulated extract during the storage period. Moreover, 0.5% encapsulated TE reduced the population of E. coli O157:H7 below the acceptable level (< 2) from Day 9 until the end of the storage period. In conclusion, the results obtained from this study suggest that liposomal encapsulation may help to obtain higher antimicrobial activity in lower TE concentrations in minced fish.     

发酵豆粕替代鱼粉对斑点叉尾生长和饲料表观消化率的影响

以发酵豆粕25%、50%、75%和100%的比例分别等量替代基础日粮中的鱼粉,研究了发酵豆粕对斑点叉尾(Ictalurus punctatus)生长和饲料表观消化率的影响。结果显示:以25%～75%的发酵豆粕替代鱼粉时,斑点叉尾增重率、特定生长率、饲料干物质和粗蛋白的表观消化率比对照组稍高(P>0.05),饲料系数与对照组没有显著性差异(P>0.05);而以发酵豆粕100%的比例替代鱼粉时,上述5种指标与对照组无显著性差异。结果表明,斑点叉尾日粮中发酵豆粕可以100%替代鱼粉,其中以25%的比例替代效果更佳。     

花生四烯酸对雨生红球藻细胞生长和虾青素含量的影响

研究了不同质量浓度花生四烯酸对雨生红球藻细胞生长和虾青素含量的影响。结果表明,适宜质量浓度的花生四烯酸(62.5 mg/L)能促进雨生红球藻细胞的生长,而低质量浓度(0.1~12.5 mg/L)和高质量浓度(312.5~1562.5 mg/L)的花生四烯酸均能抑制雨生红球藻细胞的生长;花生四烯酸能促进雨生红球藻虾青素的积累,随着花生四烯酸质量浓度的增加,雨生红球藻虾青素含量也逐渐增加,当花生四烯酸质量浓度为312.5 mg/L时,虾青素的含量最高,达3.67 mg/L,比对照组提高48.8%。     

The effect of non‐marine HUFA supplementation with fish oil removal on growth and survival of the Pacific white shrimp,Litopenaeus vannamei

The use of non‐marine arachidonic acid (ArA) and docosahexaenoic acid (DHA) as highly unsaturated fatty acid (HUFA) enrichments was evaluated as complete replacements for marine fish oil in practical diets formulated with solvent‐extracted soybean meal (SESM). Litopenaeus vannamei juveniles (0.59 g) were reared over 84 days in an outdoor tank system with no water discharge. Fishmeal was replaced with SESM, while fish oil was replaced with HUFA‐rich algal cells, alternative oil and/or fermentation products. Spray‐dried Schizochytrium algal cells (Schizomeal‐Hi DHA) served as the DHA enrichment source. Oil extracted from Mortierella sp. was used as the ArA enrichment (AquaGrow^® ArA). DHA and ArA sources (Advanced BioNutrition Corp., Columbia, MD, USA) were non‐marine products obtained from a commercial supplier. Five diets were formulated with ArA inclusion levels of 0, 0.65, 1.3, 2.6 and 5.2 g kg^?1. In addition, one diet was formulated to be DHA deficient and another was formulated with menhaden fish oil (control). Different inclusion levels of non‐marine ArA had no effect on survival or growth. Shrimp fed the non‐marine HUFA‐supplemented diets had lower average weight compared to shrimp offered the diet containing fish oil. No differences were detected in average weights of shrimp offered the ArA‐deficient and ArA‐supplemented diets.     

Effects of dietary probiotic Lactobacillus plantarum and whey protein concentrate on the productive parameters,immunity response and susceptibility of Nile tilapia,Oreochromis niloticus (L.), to Aeromonas sobria infection

Immunostimulatory feed supplements have an increasingly interest in aquaculture management. Generally, an individual supplement was used in fish diets but it is expected that the use of multi‐supplements may show synergistic enhancements in fish performance, health, and immunity. Therefore, the present investigation was carried out to evaluate the use of dietary probiotic Lactobacillus plantarum and whey protein concentrate (WPC) in practical diets for Nile tilapia, Oreochromis niloticus. Hence, probiotic L. plantarum, WPC and their mixture were incorporated into a basal fish diet (300 g/kg crude protein) as follows: T1 = a basal control diet, T2 = a basal diet containing L. plantarum, T3 = a basal diet containing 1.0 g WCP/kg diet and T4, T5 or T6 = basal diets containing probiotic L. plantarum + 1.0, 2.0 or 3.0 g WCP/kg diet, respectively. Fish (15.2 ± 0.6 g) were fed on one of the tested diets up to apparent satiation twice a day for 60 days. After that, fish were intraperitoneally injected with pathogenic bacteria Aeromonas sobria and fish mortality was observed for 10 days postchallenge. Fish growth and feed intake were significantly improved by dietary probiotic L. plantarum (T2) and/or WPC (T3) over the control group (T1), and highest fish performance was observed in T5–T6 fish groups. Similarly, highest values of haematocrit, glucose, total proteins, albumin, and globulin were significantly observed in T5–T6 fish groups. Likewise, fish fed dietary probiotic L. plantarum (T2), WPC (T3), and their mixture (T4–T6) showed antioxidants and immune‐stimulating activities better than the control group. Fish fed the control diet were more susceptible to A. sobria infection showing highest fish mortality (75.0%). Meanwhile, dietary probiotic L. plantarum (T2), WPC (T3), and their mixture (T4–T6) enhanced significantly the fish resistance to A. sobria infection resulting in maximum values of relative percent of fish survival (73.3%–80.0%) in T5–T6 groups. The present investigation recommended the use of probiotic L. plantarum with 2.0 g WPC/kg diet to improve the growth, antioxidant, immunity responses and tolerance of Nile tilapia to A. sobria infection.     

饲料中菜籽油替代鱼油对大黄鱼生长、肌肉脂肪酸组成和体色的影响

以初始体质量为(13.56±0.05)g的大黄鱼为对象,研究饲料中菜籽油替代鱼油对大黄鱼生长、肌肉脂肪酸组成和体色的影响。以鱼油组为对照组,用菜籽油分别替代25%、50%、75%、100%的鱼油,配制5种等氮等脂的实验饲料,在海水浮式网箱中进行为期8周的摄食生长实验。结果显示,饲料中不同水平菜籽油替代鱼油对大黄鱼的存活率(SR)和特定生长率(SGR)均无显著性影响;但饲料系数(FCR)随着替代水平的增加呈上升趋势,在100%替代组显著高于对照组。各处理组之间全鱼粗蛋白质、粗脂肪、灰分和水分含量均无显著性差异。肌肉中脂肪酸和饲料中脂肪酸线性关系分析表明,随着菜籽油替代鱼油水平的升高,大黄鱼肌肉中C18∶0、C18∶1、C18∶2n-6和C18∶3n-3含量不断增加,而C20∶4n-6和C22∶5n-3含量不断降低。肌肉中的饱和脂肪酸(SFA)含量随着替代水平的升高而升高,与饲料中SFA含量的变化趋势相反。菜籽油替代鱼油对大黄鱼腹部皮肤亮度值(L*)无显著性影响,但显著影响了背部皮肤L*值,100%替代组L*值显著高于对照组。对照组腹部皮肤红色值(a*)显著高于替代组;与腹部皮肤红色值相反,对照组背部皮肤红色值低于各替代组。菜籽油替代鱼油对大黄鱼背部和腹部皮肤黄色值(b*)均无显著性影响。研究表明,在本实验条件下,菜籽油替代鱼油对大黄鱼生长和体组成无显著影响,却显著影响了大黄鱼的肌肉脂肪酸组成和体色。     

Effects of substituting dietary fish oil with crude palm oil and palm fatty acid distillate on growth,muscle fatty acid composition and the activities of hepatic lipogenic enzymes in snakehead (Channa striatus,Bloch 1793) fingerling

Three diets were formulated to be iso‐nitrogenous (450 g kg^?1), iso‐lipidic (65 g kg^?1) and iso‐energetic (18.5 KJ g^?1), varying only in their lipid sources and designated as 100% fish oil (FO), 100% crude palm oil (CPO) and 100% palm fatty acid distillate (PFAD). Feed were hand fed to homogenous groups of 12 Channa striatus fingerlings (mean weight 3.5 ± 0.3 g) per tank in triplicate for 12 weeks, in a recirculation system. The growth performance and feed intake in the CPO and PFAD treatments were significantly (P<0.05) higher than those in the fish fed the control diet (FO), respectively, whereas the feed conversion ratio was better in PFAD than that in the other treatments respectively. The biological indices monitored (hepatosomatic index and viscerosomatic index) as well as carcass yield did not vary significantly among all the treatments respectively. The muscle fatty acid (FA) profile of fish was influenced by the composition of the diets fed, whereas no differences were recorded in the activities of the hepatic lipogenic enzymes monitored (fatty acid synthetase, citrate cleavage enzyme and malic enzyme). Whole‐body proximate composition analysis revealed that PFAD treatment, compared with others, contained significantly higher protein and ash, but lower lipid contents, although the muscle content of these nutrients was similar among all the treatments. Based on the results of this trial, CPO and PFAD could be used to partially substitute FO in the diet for C. striatus fingerling, to achieve good growth performance without any negative effects or compromising the muscle n‐3 FA composition (especially in the docosa hexaenoic acid and eicosa pentaenoic acid content).