Purification and characterisation of immunoglobulins from the serum of a catfish, Clarias gariepinus (Burchell, 1822)

Attempts have been made to characterize and purify immunoglobulins from the serum of Clarias gariepinus, which has been immunized with bovine serum albumen. Initially, the proteins in the serum were chromatographed successively by affinity chromatography column. The affinity-purified fraction was concentrated and checked in SDS-PAGE, two bands of heavy chain and two bands of light chain were observed. Since teleost immunoglobulins have been shown to belong to a single class, the extra bands in light and heavy chains in the present study might be the breakdown of immunoglobulin or some unpurified contaminants. The affinity-purified fraction was also subjected to gel filtration chromatography column.

Flavour components of some processed. fish and fishery products of Japan

A study was conducted to examine the flavour components of some processed fish and fishery products of Japan by gas chromatography-mass spectrometry (GC-MS). In brief the method was to absorb the headspace volatiles at 70°C into the fused silica fibre of needle of the solid phase micro extraction fibre. The absorbed components were injected to the GC-MS. The components were identified by computer matching with library database as well as by authentic standard components. In general the number of flavour components were higher in the processed fish and fishery products (except frozen prawn) than that of the raw fish and prawn. The concentration (quantity) of the f1avour components in processed fish and fishery products was much higher than that of the raw fish and prawn. Smoked salmon and baked salmon possessed double number of flavour components than that of the raw salmon. Smoking resulted the highest number of flavour components followed by baking (grilling) and canning, surimi products (kamaboko and chikuwa), drying and lastly salting. However, freezing and frozen storage resulted loss of flavour components in prawn.

The influence of plating technique and incubation temperature on bacterial count from fish and fishery products

For bacterial sampling of raw unprocessed fish and frozen fishery products, spread plate method is preferable to pour plate method; incubation of plates at 30°C gives a higher count than incubation at 37°C. Analysis of variance of the data shows that sample variation between different types of fishes is highly significant whereas the variations between triplicate plates are not significant at 5 % level.

The effect of washing on the quality of minced catfish during frozen storage

The effect of washing minced catfish in water, sodium chloride solution (1%) and ascorbic acid solution (0.1%) in improving the quality and frozen shelf-life has been studied. Washing improved the colour and reduced the non-protein nitrogen contents and extractable nitrogen. Denaturation was more in samples washed in salt and ascorbic acid solutions. Rancidity as measured by PV and organoleptic studies showed significant reduction in washed samples. The frozen storage life was significantly enhanced by washing.

Occurrence of enterotoxigenic staphylococci in frozen fishery products

Sixty six samples of frozen fishery products consisting of frozen crab meat, frozen prawns, frozen cuttlefish and frozen mussel collected from nine cold stores situated in and around Cochin city were examined for coagulase positive staphylococci. Forty eight samples (72.72%) revealed the presence of coagulase positive staphylococci, which were then tested for enterotoxigenity, employing cellophane-over-agar method for toxin production and optimum-sensitivity-plate (OSP) method for toxin detection. Seventy seven out of one hundred and two (75.49%) coagulase positive staphylococcal isolates from the above products produced enterotoxins A, B, C, D and E either singly or in combinations. Enterotoxin A and D were detected more often than others. The occurrence of enterotoxigenic staphylococci in frozen fishery product is reported for the first time in this country.

Effect of liquid nitrogen freezing and subsequent storage on survival of Staphylococcus aureus and Streptococcus pyogenes in treated prawn meat

Prawn meat treated with Streptococcus pyogenes B-49-2 culture and Staphylococcus aureus ATCC-12598 culture were frozen in conventional plate freezer at -40°C and by spray type liquid nitrogen freezer. The frozen products were stored at -18°C. Streptococcus pyogenes B-49-2 showed low sensitivity to cold injury during freezing and frozen storage. Staphylococcus aureus ATCC-12598 survived during the entire storage period of 240 days. Total bacterial count of untreated prawn meat was found to be always lesser in liquid nitrogen frozen products than that in plate frozen products.

Incidence and low temperature survival of coagulase positive staphylococci in fishery products

Coagulase-positive staphylococci was found to be absent in all the frozen samples of lobsters, cuttle fish, cat fish, seer fish and red snapper examined. Coagulase-positive staphylococci were present in 38% of the cooked frozen shrimps and only 16% of the samples had staphylococci count more than 100/g. In the case of headless, peeled and deveined, peeled undeveined shrimps, the incidence of the organism was 6, 12 and 16% respectively. The study indicated that the incidence of coagulase-positive staphylococci is not a serious problem in frozen fishery products processed in this country. There was remarkable difference in the rate of destruction of coagulase-positive staphylococci in raw and cooked shrimps during freezing and frozen storage.

Effects of the quick and slow freezing on the quality of Nile Tilapia fillets (Oreochromis niloticus) and Red Tilapia fillets (O. niloticus and Tilapia mosambicus)

In this study, quality of fresh, slow frozen and quick frozen tilapia fillets and its changes during storage at -18C° were investigated. For preparation the samples, fresh tilapia fillets were frozen by slow and quick frozen methods. Slow frozen samples were prepared by storing the packed fillets directly in the -18 C°. The sprila freezing tunle with -30C° was also used for preparation the quick frozen sample. The quick frozen samples were then stored at -18C°for six months. Proximate composition, fatty acid profiles, TBA, PV, TVN, Total cuont, Drip loss, and sensory evaluation of the samples were determined in every month. Scanning Electron Microscopy (SEM) was used for study on the effects of the frozen condition on the microstructure of the fillets. Results indicated that two different frozen methods had significantly different effects on the quality of the fillets. Most of the proximate composition (protein, moistre and fat) reduced during the storage. Quick frozen filets had significantly (P<0.05) lower reduction than slow frozen samples. All of the chemical quality indexes (PV, TBA, and TVN) increased during the storage as compered to the fresh samples. In these paramethers, the slow freezing had higher changes than quick freezing metods (P<0.05). The microbial properties of the samples showed decrese during the storage. Lower amont of total cuont was observed at the end of the storage time in the quick frozen samples than slow frozen once (P<0.05). The large changes in the fatty acid profiles of the sample were fond in all samples. During the storage SFA and MUF of the samples increased however, the PUFA decresed. A lower change was obseved in the quick frozen samples than slow frozen samples (P<0.05). Drip loss was increased in both frozen samples during the storage period. The percentage of the drip in the slow frozen samples was significantly higer than quick frozen samples (P<0.05). SEM micrographs were also showed that the chnges in the microstructur of the samples was different in the slow and frozen samples. Slow freezing methods had higher damge in the microstructure of the sample then quick freezing mathods. Sensory evaluation of the samples indicated that a better acceptability in the quick frozen samples than slow frozen sample (P<0.05).

Extraction and structure elucidation of natural compounds of some gorgonians of the Persian Gulf and also physiological and pharmacological effects of gorgonian extract on some selected organisms

In the present research, investigations were carried out for structure elucidation of natural compounds and also for studing biological and teratogenical effects of two Genus of soft corals named as " Echinogorgia cf. indica" and "Sinularia erecta" in Persian Gulf. First, 350 gr Echinogorgia was extracted by Acetone, then, the extract was separated by ether from aqueos phase to give 4.5 gr oil. The oil eluted with Petrol - ether Et2 o (9:1) which was recovered Linderazulene and it's derivative as purple Cristals (350 mg/ca 0.1 %). In order to determine molecular structure, the Samples were used for spectroscopic method as: H1- NMR , C13- NMR and 2D NMR. Also, for extraction and structure elucidation of natural compounds, the soft coral " sinularia erecta " were used 1187/37 gr and extracted by Aceton. The extract was concentrated and resulting aqueous suspension and extracted by using ether to give 8.41 gr oil. The oil , was Chromatographed on a column of silica gel and some different fractions were gathered. Initial fraction (1-11) which were nonpolar compounds were seprated by GC/MS. Mass spectrum were prepared and much compounds were recognized.

微生物法生成辅酶Q10的研究

本文根据我们实验室建立的发酵产物中辅酶Q10定性定量检测方法，筛选得到一株可以代谢产生较多辅酶Q10的野生菌株放射形土壤杆菌(Agrobacterium radiobacter No.50)。 为了提高放射形土壤杆菌的辅酶Q10的产量，本实验利用液体培养研究了单因素对菌株辅酶Q10产量的影响，并用正交法确定了最佳液态发酵条件。最佳发酵培养基是：葡萄糖20g,蔗糖40g, 硫酸铵10g,玉米浆30g, 酵母膏3g，K2HPO4 3g,MgSO4.7H2O 1g，蒸馏水1000mL,pH 7.0-7.2。最佳发酵条件是：转接斜面菌种到种子培养基, 转速220r/min、温度28。C培养24h后，转入发酵培养基(250mL三角拼装液量为50mL，pH 7.0), 接种量为10％，转速220r/min、温度28。C，培养120h。在此条件下，菌体湿重约为50g/L，辅酶Q10含量约为20mg/L。 本文以放射形土壤杆菌为出发菌株进行诱变育种，以期获得辅酶Q10高产菌。根据微生物育种原理、参照辅酶Q10的代谢调控机制，以野生型放射形土壤杆菌(Agrobacterium radiobacter No.50)为出发菌株，采用紫外线和亚硝基胍复合诱变技术，依次筛选得到菌体提取物M抗性菌ARM－7、烟草提取物T抗性菌株ARMT-26、Vk3抗性菌株ARMTV-25、链霉素抗性菌株ARMTVS-32，菌株ARMTVS-32产量达到了36.8mg/L，与原始出发菌株相比，产量提高了77％。 研究了茄尼醇、对羟基苯甲酸、橘子皮提取物D、胡萝卜提取物E、烟草提取物对ARMTVS-32合成辅酶Q10的影响，结果表明这些物质对菌体合成辅酶Q10有一定促进作用，添加0.2g/L茄尼醇时，辅酶Q10含量提高了17%，达到了40.7mg/L；添加1.2g/L橘子皮提取物D时，辅酶Q10含量提高了13.8%，达到了39.6mg/L；添加0.5g/L胡萝卜提取物E时，辅酶Q10含量提高了25.3% ，达到了43.6mg/L；添加8g/L烟草提取物时，辅酶Q10含量提高了12.6%，达到了39.2mg/L。 Production of Coenzyme- Q10 (CoQ10) by fermentation is considered as a process with broad prospects.Quantitative Analysis of CoQ10 in the culture of microbe by TLC—UV spectrophotometry was developed, by using this method we got the strain Agrobacterium radiobacter,which was isolated from forest soil of southwest of China. The effect of the single factor on CoQ10-production ability of the strain was examined by liquid cultured, and its best optimum cultivation conditions were established by orthogonal method. The results showed that the optimum fermentation conditions were as following: carbon sources glucose 20g/L,sucrose 40g/L; nitrongen sources (NH4)2SO4 10g/L,maize liquid 30g/L;yeast extract 3g; K2HPO4 3g/L,MgSO4.7H2O 1g/L; initial pH was 7 and volume of medium(medium volume vs flask volume) was 50mL/500mL, incubating for 120h on a rotary shaker at 220 rpm and 28℃.Under these conditions, the biomass and CoQ10 concentration reached 50g/L and 20mg/L respectively. According to the biosynthesis mechanism of CoQ10 and breeding theory, CoQ10 over-production strains were screened by UV--NTG. mutation using Agrobacterium radiobacter No.50 as parent strain. A microbe-juice resistant mutant ARMTVS-32, which also could resist tobacco-juice, VK3 and streptomycin, was screened out from an agar plate. The CoQ10 content of ARMTVS-32 reached 36.8mg/L, which was 77% higher than the initial strain. In addition, We discussed the effects of some organic substrates on the synthesis of CoQ10 in ARMTVS-32. The results showed that solanesol, orange juice D, carrot juice E and tobacco juice could promote the CoQ10 accumulation in the cells. The CoQ10 content of ARMTVS-32 reached 40.7mg/L when added 0.2g/L solanesol,it reached 39.6mg/L when added 1.2g/L orange juice D，it reached 43.6mg/L when added 0.5g/L carrot juice E. it reached 39.2mg/L when added 8g/L tobacco juice.

A STUDY ON THE LUMINESCENCE PROPERTIES OF EU3+ AND DY3+ IN M(2)RE(8)(SIO4)(6)O-2 (M=MG, CA RE=Y, GD, LA)

In this paper, the luminescence properties of Eu3+ and Dy3+ in the oxyapatites M(2)RE(8)(SiO4)(6)O-2 (M=Mg, Ca; RE=Y, Gd, La) were studied. The spectral characters of Eu3+ were discussed in relation to the crystal structure. The dependence of the red-to-orange intensity ratio and the position of the charge transfer band of Eu3+ and the yellow-to-blue intensity ratio of Dy3+ together with their fluorescence intensities (I-R for Eu3+ and I-Y for Dy3+) On the M(2+) and the substitution of BO45- and PO43- for SiO44- was discussed. Finally, the concentration quenching of Dy3+ luminescence was reported.

A NOVEL SHELL COLOR VARIANT OF THE PACIFIC ABALONE HALIOTIS DISCUS HANNAI INO SUBJECT TO GENETIC CONTROL AND DIETARY INFLUENCE

Molluscan shells may display a variety of colors, which formation, inheritance, and evolutionary significance are not Well understood. Here we report a new variant of the Pacific abalone Haliotis discus hannai that displays a novel orange shell coloration (O-type) that is clearly distinguishable from the Wild green-shelled abalone (G-type). Controlled mating experiments between O- and G-type abalones demonstrated apparent Mendelian segregations (1:1 or 3:1) in shell colors in F-2 families, which support the notion that the O- and G-types are under strict genetic control at a single locus With a recessive o (for orange shell) allele and a dominant G (for green shell) allele. Feeding with different diets caused modifications of shell color within each genotype, ranging from orange to yellow for O-type and green to dark-brown for the G-type, without affecting the distinction between genotypes. A previously described bluish-purple (B-type) shell color was found in one of the putative oo X oG crosses, suggesting that the B-type may be it recessive allele belonging to the same locus. The new O-type variant had no effect on the growth of Pacific abalone on the early seed-stage. This Study demonstrates that shell color in Pacific abalone is subject to genetic control as well as dietary modification, and the latter probably offers selective advantages in camouflage and predator avoidance.

Market-oriented new product development of functional beverages

Strategic reviews of the Irish Food and Beverage Industry have consistently emphasised the need for food and beverage firms to improve their innovation and marketing capabilities, in order to maintain competitiveness in both domestic and overseas markets. In particular, the functional food and beverages market has been singled out as an extremely important emerging market, which Irish firms could benefit from through an increased technological and market orientation. Although health and wellness have been the most significant drivers of new product development (NPD) in recent years, failure rates for new functional foods and beverages have been reportedly high. In that context, researchers in the US, UK, Denmark and Ireland have reported a marked divergence between NPD practices within food and beverage firms and normative advice for successful product development. The high reported failure rates for new functional foods and beverages suggest a failure to manage customer knowledge effectively, as well as a lack of knowledge management between functional disciplines involved in the NPD process. This research explored the concept of managing customer knowledge at the early stages of the NPD process, and applied it to the development of a range of functional beverages, through the use of advanced concept optimisation research techniques, which provided for a more market-oriented approach to new food product development. A sequential exploratory research design strategy using mixed research methods was chosen for this study. First, the qualitative element of this research investigated customers’ choice motives for orange juice and soft drinks, and explored their attitudes and perceptions towards a range of new functional beverage concepts through a combination of 15 in-depth interviews and 3 focus groups. Second, the quantitative element of this research consisted of 3 conjoint-based questionnaires administered to 400 different customers in each study in order to model their purchase preferences for chilled nutrient-enriched and probiotic orange juices, and stimulant soft drinks. The in-depth interviews identified the key product design attributes that influenced customers’ choice motives for orange juice. The focus group discussions revealed that groups of customers were negative towards the addition of certain functional ingredients to natural foods and beverages. K-means cluster analysis was used to quantitatively identify segments of customers with similar preferences for chilled nutrient-enriched and probiotic orange juices, and stimulant soft drinks. Overall, advanced concept optimisation research methods facilitate the integration of the customer at the early stages of the NPD process, which promotes a multi-disciplinary approach to new food product design. This research illustrated how advanced concept optimisation research methods could contribute towards effective and efficient knowledge management in the new food product development process.

Characterisation and application of fruit by-products as novel ingredients in gluten-free products

The physicochemical and nutritional properties of two fruit by-products were initially studied. Apple pomace (AP) contained a high level of fibre and pectin. The isolated AP pectin had a high level of methylation which developed viscous pastes. Orange pomace also had high levels of fibre and pectin, and it was an abundant source of minerals such as potassium and magnesium. Due to the fibrous properties of orange pomace flour, proofing and water addition were studied in a bread formulation. When added at levels greater than 6%, the loaf volume decreased. An optimised formulation and proofing time was derived using the optimisation tool; these consisted of 5.5% orange pomace, 94.6% water inclusion and with 49 minutes proofing. These optimised parameters doubled the total dietary fibre content of the bread compared to the original control. Pasting results showed how orange pomace inclusions reduced the final viscosity of the batter, reducing the occurrence of starch gelatinisation. Rheological properties i.e. the storage modulus (G') and complex modulus (G*) increased in the orange pomace batter compared to the control batter. This demonstrates how the orange pomace as an ingredient improved the robustness of the formulation. Sensory panellists scored the orange pomace bread comparably to the control bread. Milled apple pomace was studied as a potential novel ingredient in an extruded snack. Parameters studied included apple pomace addition, die head temperature and screw speed. As screw speed increased the favourable extrudate characteristics such as radical expansion ratio, porosity and specific volume decreased. The inclusion of apple pomace had a negative effect on extrudate characteristics at levels greater than 8% addition. Including apple pomace reduced the hardness and increased the crispiness of the snack. The optimised and validated formulation and extrusion process contained the following parameters: 7.7% apple pomace, 150°C die head temperature and a screw speed of 69 rpm.

Intracellular Cytokine Staining and Flow Cytometry: Considerations for Application in Clinical Trials of Novel Tuberculosis Vaccines.

Intracellular cytokine staining combined with flow cytometry is one of a number of assays designed to assess T-cell immune responses. It has the specific advantage of enabling the simultaneous assessment of multiple phenotypic, differentiation and functional parameters pertaining to responding T-cells, most notably, the expression of multiple effector cytokines. These attributes make the technique particularly suitable for the assessment of T-cell immune responses induced by novel tuberculosis vaccines in clinical trials. However, depending upon the particular nature of a given vaccine and trial setting, there are approaches that may be taken at different stages of the assay that are more suitable than other alternatives. In this paper, the Tuberculosis Vaccine Initiative (TBVI) TB Biomarker Working group reports on efforts to assess the conditions that will determine when particular assay approaches should be employed. We have found that choices relating to the use of fresh whole blood or peripheral blood mononuclear cells (PBMC) and frozen PBMC; use of serum-containing or serum-free medium; length of stimulation period and use of co-stimulatory antibodies can all affect the sensitivity of intracellular cytokine assays. In the case of sample material, frozen PBMC, despite some loss of sensitivity, may be more advantageous for batch analysis. We also recommend that for multi-site studies, common antibody panels, gating strategies and analysis approaches should be employed for better comparability.