Fatty acid composition of more brown seaweeds (Phaeophyta) from the coast of Karachi

Six species of multicellular, thalloid, brown algae, belonging to five genera of the classes Laminariophyceae and Fucophyceae were collected from the coastal areas of Buleji near Karachi (Pakistan) during October 1997 and February 1998. They were extracted in chloroform: methanol, saponified, subjected to column chromatography (CC, TLC), esterified and analysed for fatty acid (FA) composition initially by gas-liquid-chromatography (GLC) and finally by gas chromatography-mass spectrometry (GC-MS). They displayed only a few SCFAs, PUFAs and substituted FAs, no VLCFA, C22 UFA, CFA, DCFA and monoynoic FA, large amount of CI6:0, very large quantity of C18:1, very small RCCL and FA-diversity, C18 UFAs up to four DBs, C20 UFAs up to three DBs only. They were characterized by the largest amount of C18:1, lowest degree of unsaturation of C20 UFAs, lack of C22 UFAs, the shortest RCCL and the smallest FA-diversity as compared to other phyla.

Application of encapsulated nisin and sodium citrate for shelf life increasing of Rainbow trout (Onchorhynchus mykiss) fillet packaged with MAP and their effect on Staphylococcus aureus

Nisin is a widely used naturally occurring antimicrobial effective against many pathogenic and spoilage microorganisms. It has been proposed that reduced efficacy of nisin in foods can be improved by technologies such as encapsulation to protect it from interferences by food matrix components. The aim of this study was using of spray dried encapsulated nisin with zein in concentration of (0.15 and 0.25 g/kg) and sodium citrate (1.5 and 2.5%) and treatments with both of them to extent the shelf life of filleted trouts packaged by Modified Atmosphere Packaging (45% CO2, 50% N2 ,5% O2) and stored at 4±1 °C for 20 days. Furthermore, to evaluate the antimicrobial efficiency of encapsulated nisin and soudium citrate the trouts fillets was inoculated with Staphylococcus aureus as an index pathogenic bacteria. Assessment of chemical spoilage indexes such as (Proxide value, Thiobarbituric acid, total volatile base nitrogen and pH) , microbial parameters (Total Plate Count, Psychrotrophic count, Lactic acid bacteria count), Staphylococcus aureus cont in treatments which were inoculated with 5 logcfu/g of this bacteria and sensory evaluation of fillets including (smell, color, texture and total acceptability) was carried out in days of 0, 4, 8, 12, 16 and 20. The results revealed that treatment with both exposure of nisin and sodium citrate showed significantly lower chemical spoilage indexes in comparison with controls (vaccum packed and MAP) (P<0.05). Furthermore, (nisin 0.25 g/kg sodium citrate 2.5%) treatment which was exposed to the maximal level used of both materials was significantly the lowest treatment with (Proxide value, Thiobarbituric acid, total volatile base nitrogen and pH) of 9.95 (meq O2/kg) , 1.55 (mgMA/kg), 29.65 (mgN/100g) and 6.65 , respectively and according to the maximal recommended level of this indices , shelf life of fillets in this treatment was esstimated 20 days.The control (vaccum packed) treatment was significantly the highest treatment with (Proxide value, Thiobarbituric acid, total volatile base nitrogen and pH) of 15.17 (meq O2/kg), 3.03 (mgMA/kg), 38.4 (mgN/100g) and 6.95 , respectively and according to the maximal recommended level of this indices , shelf life of fillets in this treatment was estimated 11 days. Also, in microbial point of view (nisin 0.25 g/kg- sodium citrate 2.5%) treatment was the lowest treatment with Total Plate Count, Psychrotrophic count, Lactic acid bacteria count and Staphylococcus aureus count of 6.7, 6.83, 5.25 and 6.04 logcfu/g respectively, and conrol (vaccum packed) treatment was the highest treatment with 9.15, 9.41, 7.7 and 9.01 logcfu/g respectively. According to the lower results of chemical and microbial indices and higher sensory evaluated scores assessed in this research for encapsulated nisin in comparison with free nisin , it was concluded that encapsulation of nisin with zein capsules may improve the efficiency of nisin. The measuremented values of Mass yield, Total solids content of capsules, Encapsulation efficiency, In vitro release kinetics in 200 hour for encapsulated nisin in this study was 49.89, 62, 98.31 and 69% respectively and Encapsulated particle size was lower than 674.21 μm for 90% of particles. As a consequence, nisin , in particular encapsulated nisin, and sodium citrate alone or together with and Modified Atmosphere packaging might be considered as effective tools in preventing the quality degradation of the fillets, resulting in an extension of their shelf life.

A study on the effect of Zataria multiflora Bioss essential oil and Nisin on the growth of Lactococcus garvieae in the rainbow trout fillet (Oncorhynchus mykiss)

The ever-increasing population of the world and the growing need for animal protein has doubled the modern man’s demand for food. Additionally, the improvement in the general public health, and the worsening of environmental/ecological pollution have prompted today’s world to look for ways to procure healthy food. And one such attempt is the use of natural preservatives to decrease the bacterial load in foodstuffs, in other words, to increase their durability. This study evaluates the effects of different concentrations of Zataria multiflora Bioss (EO 0, 0.005, 0.015, 0.045, 0.135, 0.405%) and Nisin (0, 0.25, 0.5, 0.75 μg/ml) and storage time (9 days) on the growth of Lactococcus garvieae Ir-170A(856bp) alone, and their combination in a food model system (fillets of the rainbow trout (Oncorhynchus mykiss). Additionally, the growth of a sample of this bacteria in laboratory conditions was studied. The results of this study showed that different concentrations of Nisin had a significant impact (p<0.05) on Lactococcus garvieae. With the value of t in 0.75 μg/ml, the effectiveness rose to 65.77%; the biggest effect on Lactococcus garvieae. And the effect at 4 0C exceeded 80C. The study has also demonstrated that all concentrations of Zataria multiflora Bioss were effective against Lactococcus garvieae. However, with the value of t at 0.405%, the effectiveness was 71.91%. This value had the biggest effect on Lactococcus garvieae. At 4 0C, the effect surpassed the one at 80C. The synergistic effects of the EO and Nisin showed that with the value of t at 0.405% EO and 0.75 μg/ml Nisin was 14.62% had the greatest effect on Lactococcus garvieae. In this study, multi-factorial effects for different concentrations of Zataria multiflora Bioss (EO 0, 0.005, 0.015, 0.0025%), three different concentrations of 122 Nisin (0, 0.25,0.75 μg/ml) and two different levels of PH (5.5 , 7) at two incubation temperatures (15,37) on logp% of Lactococcus garvieae during 43 days in BHI broth were evaluated. Most of the effects on Lactococcus garvieae occurred in PH 5.5 and at a temperature of 150C.

Study of effect Rosmarinus officinalis oil and nisin on the growth of Streptococcus iniae in lab conditions and fillets of rainbow trout (Oncorhynchus mykiss)

Fish are an important part of a healthy diet since they contain high quality protein, but typically present a low fat percent when compared to other meats. Fish is an extremely perishable food commodity. On the other hand, food borne diseases are still a major problem in the world, even in well-developed countries. The increasing incidence of food borne diseases coupled with the resultant social and economic implications means there is a constant striving to produce safer food and to develop new antimicrobial agents concerns over the safety of some chemical preservatives and negative consumer reactions to preservatives they perceive as chemical and artificial, have prompted on increased interest in more ‘‘naturalgreen’’ alternatives for the maintenance or extension of product shelf-life. Particular interest has focused on the potential applications of plant essential oils. However, to establish the usefulness of natural antimicrobial preservatives, they must be evaluated alone and in combination with other preservation factors to determine whether there are synergistic effects and multiple hurdles can be devised. In this study, were evaluated the effects of different concentrations of Rosmarinus officinalis and nisin and storage time (15 days) on growth of Streptococcus iniae GQ850377 in a lab conditions and a food model system (fillets of rainbow trout) in 4 and 8 °C. In addition, we also studied multi factorial effects of four different concentration of rosemary, three different concentrations of nisin, two different levels of pH in 3 temperature 4,15 and 37 °C on log% of S.iniae during 43 days in BHI broth. The results on growth of S. iniae were evaluated using SPSS 20.0 statistical software and analyzed the logarithm of total count of the bacterial by Tukey Test. Results were considered statistically significant when P<0.05. MIC and MBC values of rosemary and nisin were 0.03, 0.075 % and 5, 40 μg/mL, respectively. The growth of S. iniae was effected significantly (P<0.05) by rosemary and nisin and also combination of rosemary and nisin in 4 and 8 °C. Samples treated with 0.135 and 0.405 % of rosemary showed a significant decrease on the growth of the bacteria compared with control sample(P<0.05). The most ١٤٦ inhibitory effects were seen in samples treated with 0.135 and 0.405% of rosemary until 9 days after storage. Also, the synergism effects of rosemary and nisin on the growth rate of bacteria was significant (P<0.05) compared with untreated samples and samples treated with the rosemary or nisin, only. Synergistic effects was observed at concentration of 0.405% rosemary and 0.75 μg/mL nisin in both temprature. Results of this study showed that different concentration of rosemary a significant inhibitory effect (P<0.05) on log% of S. iniae, in BHI broth in pH 5.5 and 7 in 4,15 and 37 °C during 43 days. In concentration of 0% rosemary (control) in pH 5.5 and 7 and 37°C, log% were 1.099 and 3.15, whereas in concentration of 0.015% rosemary were -4/241 and 1.454, respectively. The use of essential oils may improve food safety and overall microbial quality. If essential oils were to be more widely applied as antibacterials in foods, the organoleptic impact would be important. In addition, it is recommended to apply essential oils or their compounds as part of a hurdle system and to use it as an antimicrobial component along with other preservation techniques. Thus essential of R. officinalis with high antibacterial activity selected in this study could be a potential source for inhibitory substances against some food-borne pathogens and they may be candidates for using in foods or food-processing systems.

Effect of thawing methods on the chemical, biochemical, microbiological and sensory indices of Caspian Sea kutum (Rutilus frisii kutum) and rainbow trout (Oncorhynchus mykiss)

Effects of different thawing method i.e. in a refrigerator, in water, at air ambient temperature and in a microwave oven on proximate, chemical (PV, TBA, FFA, TVB-N, SSP, FA), biochemical (pH, WHC,ThL), microbial (total viable, psychrotrophic, coliform, Shewanella and yeast-mould count) and sensory analysis were carried out on frozen whole Caspian sea Kutum (Rutilus frisii kutum) and Rainbow trout (Oncorhynchus mykiss) carcasses. The values of ash, protein, SSP, WHC, PUFA, PUFA/SFA. EPA+DHA/C16:0, pH, and microbial count of thawed samples decreased significantly while fat, PV, TBA, FFA, TVB-N, SFA and MUFA increased compared to the fresh fish (unfrozen) as control samples. Also, sensory evaluation all of thawed samples showed a significant (p<0.05) quality loss compared to the fresh fish as control samples. The lowest chemical and biochemical values as well as microbial growth were determined in water thawed samples. Therefore, based on this study thawing in water is most suitable for frozen whole rainbow trout.

Effect of replacement of fish meal with soybean meal plus supplementary enzymes on growth performance, survival rate and apparent digestibility of rainbow trout (Oncorhynchus mykiss)

This study was carried out to measure the effects of a supplementary multi enzyme on growth performance , survival rate and apparent protein digestibility of rainbow trout fed some diets containing different amounts of soy bean meal. Five exprimental diets with replacement of 25, 50, 75 and 100 percent of fish meal protein by soy bean meal protein were made and 0, 500 and 1000 ppm dosages of supplementary multi enzyme had used in each of them. By the means a diet with fish meal as the only source of protein has used as the control. So this study had 13 treatments. The trouts in 89.40±4.01 gr mean weight were stocked in 39 experimental fiberglass tanks in abundance of 30 fish per any tank. These specimens fed experimental diets for 8 weeks and ten of them in each tank fed same diets which added Cr2O3 to them for one more week to measure the apparent protein digestibility in them. The results shown that supplementary multi enzyme (Avizyme) which contains Protease , Amylase and Xylanase , caused increases in growth performance , survival rate and apparent protein digestibility in trouts which fed soybean meal. Also this study shown that using 1000 ppm of Avizyme in diets which containing soybean meal had the best results and the diet which contained 39 % soybean meal with this amount of enzymes, had no significant differences by the control in any of the studied factors.

Dinoflagellate "brown tides" in Alexandria, Egypt waters during 1997-1998

Charles Darwin the research ship undertook an Oceanographic Cruise in 1986, CD 86/17 of the North Arabian Sea. Sediment cores were collected between 15° and 25°N. In this study sediment cores collected from deep Indus and Oman basins (CD 1715, CD 1730, CD 1738) have been analyzed for mineralogy, water content and porosity. In general, the cores are mainly composed of clay to silt sized terrigenous and biogenic constituents. Quartz, chlorite and illite are the common minerals of Arabian Sea sediments. Porosity determined by water content of sediments has been correlated with quartz/chlorite and quartz/illite peak ratios to show a relationship between mineral composition and physical properties.

Interactions of dietary levels of protein and energy on rainbow trout (Oncorhynchus mykiss) in desert brackish water

A 3x3 factorial experiment was conducted to determine the optimum protein to energy (P/E) ratio for rainbow trout in brackish water. Three crud protein levels and three energy levels at each protein level were utilized. Diets were made in semi-purified that in all of them fish meal, casein and gelatin as the sources of protein and dextrin, starch and oil as the sources of energy were used. Each of experimental diets was fed to triplicate groups of 20 fish with an average individual weight of 81.5 g in 9 2000-1 flow trough fiberglass tanks. During this experiment water temperature, dissolved oxygen, PH and EC were 15±2°C, 6.5-8.1 mg/1, 7.7-8.6 and 25400 grills respectively. The diets were fed at a rate between 1.6-2 wet body weight% per day depended to water temperature in three equal rations and adjusted two weekly for 84 days. At each of protein levels, weight gain percent (%WG), average daily growth percent (%ADG), protein efficiency ratio (PER), apparent net protein utilization percent (%ANPU), or percent of protein deposited, specific growth rate (SGR) and condition factor (CF) were found to increase and food conversion ratio (FCR) was found to decrease with an increasing energy levels from 370 to 430 Kcal/100g. Fish fed a 35% protein, 430 Kcal/100g energy diet with a P/E ratio of 81.4 mg protein/ Kcal PFV energy, attained the best growth performance. Fat and moisture of carcass were affected by protein and energy levels of test diets while protein and ash of carcass were relatively constant in different treatments.

The search for the IFN-gamma receptor in fish: Functional and expression analysis of putative binding and signalling chains in rainbow trout Oncorhynchus mykiss

Interferons (IFNs), consisting of three major subfamilies, type I, type II (gamma) and type III (lambda) IFN, activate vertebrate antiviral defences once bound to their receptors. The three IFN subfamilies bind to different receptors, IFNAR1 and IFNAR2 for type I IFNs, IFN gamma R1 and IFN gamma R2 for type II IFN, and IL-28R1 and IL-10R2 for type III IFNs. In fish, although many types I and II IFN genes have been cloned, little is known about their receptors. In this report, two putative IFN-gamma receptor chains were identified and sequenced in rainbow trout (Oncorhynchus mykiss), and found to have many common characteristics with mammalian type II IFN receptor family members. The presented gene synteny analysis, phylogenetic tree analysis and ligand binding analysis all suggest that these molecules are the authentic IFN gamma Rs in fish. They are widely expressed in tissues, with IFN gamma R1 typically more highly expressed than IFN gamma R2. Using the trout RTG-2 cell line it was possible to show that the individual chains could be differentially modulated, with rIFN-gamma and rIL-1 beta down regulating IFN gamma R1 expression but up regulating IFN gamma R2 expression. Overexpression of the two receptor chains in RTG-2 cells revealed that the level of IFN gamma R2 transcript was crucial for responsiveness to rIFN-gamma, in terms of inducing gamma IP expression. Transfection experiments showed that the two putative receptors specifically bound to rIFN-gamma. These findings are discussed in the context of how the IFN gamma R may bind IFN-gamma in fish and the importance of the individual receptor chains to signal transduction. (c) 2009 Elsevier Ltd. All rights reserved.

First description of a novel Weissella species as an opportunistic pathogen for rainbow trout Oncorhynchus mykiss (Walbaum) in China

Six strains of Gram-positive, catalase-negative, non-motile, irregular short rod-shaped Weissella bacteria, with width and length of 0.5-0.6 and 1.2-2.7 mu m were isolated from diseased rainbow trout Oncorhynchus mykiss (Walbaum) in winter of 2007 at a commercial fishery in Jingmen, Hubei province, China. The diseased rainbow trout exhibited hemorrhage in eyes, anal region, intestine and abdomen wall, petechia of liver, some fish with hydrocele in stomach. Six isolates had identical biochemical reactions, phylogenetic analysis of 16S rDNA sequences, amplified ribosomal DNA restriction analysis (ARDRA), enzymatic profile analysis and antimicrobial susceptibility results, indicating as a single clonal outbreak. But all were different from any other validated twelve Weissella species in the term of physiological and biochemical characters. It is indicated that isolates are phylogenetically closer to Weissella halotolerans, Weissella viridescens and Weissella minor on 16S rDNA phylogenetic analysis result, than to W halotolerans and W viridescens on the result of ARDRA study and enzymatic profile analysis. Antimicrobial susceptibility testing was used to scan effective drugs for the therapy of this disease. Experimental infection assays with one isolate were conducted and pathogenicity (by intraperitoneal injection) was demonstrated in rainbow trout O. mykiss (Walbaum) and crucian carp (Carassius auratus gibelio) fingerlings. Because no Weissella was detected in fish feedstuffs and pond water, the source of this pathogen remains unknown, and Weissella isolates were regarded as an opportunistic pathogen for rainbow trout. This is the first report of Weissella strains which can cause disease of cultured fish in the world. (C) 2009 Elsevier B.V. All rights reserved.

Identification of an additional two-cysteine containing type I interferon in rainbow trout Oncorhynchus mykiss provides evidence of a major gene duplication event within this gene family in teleosts

Multiple type I interferons (IFNs) have recently been identified in salmonids, containing two or four conserved cysteines. In this work, a novel two-cysteine containing (2C) IFN gene was identified in rainbow trout. This novel trout IFN gene (termed IFN5) formed a phylogenetic group that is distinct from the other three salmonid IFN groups sequenced to date and had a close evolutionary relationship with IFNs from advanced fish species. Our data demonstrate that two subgroups are apparent within each of the 2C and 4C type I IFNs, an evolutionary outcome possibly due to two rounds of genome duplication events that have occurred within teleosts. We have examined gene expression of the trout 2C type I IFN in cultured cells following stimulation with lipopolysaccharide, phytohaemagglutinin, polyI:C or recombinant IFN, or after transfection with polyI:C. The kinetics of gene expression was also studied after viral infection. Analysis of the regulatory elements in the IFN promoter region predicted several binding sites for key transcription factors that potentially play an important role in mediating IFN5 gene expression.

The biological effects of rainbow trout (Oncorhynchus mykiss) recombinant interleukin-8

In this report, recombinant interieukin-8 (rIL-8) was produced and its activity tested for the first time in fish. The rainbow trout rIL-8 was produced in Escherichia coli and purified using a 6xHis tag at the N-terminus. The rIL-8 induced a dose-dependent migration of head kidney leukocytes at concentrations from 0.1 to 10 ng/ml, with a peak response at 1 ng/ml. Trout rIL-8 also had a significant effect on superoxide production by head kidney cells, with maximal, activity at 0.1 and 1 ng/ml. When injected intraperitoneally into trout, rIL-8 had a clear effect on total leukocyte number in the peritoneal cavity, with increasing doses (up to 5 mu g) eliciting more cells. Of three leukocyte types distinguished, neutrophils were the dominant cell type, especially at higher rIL-8 concentrations. In contrast, the proportion of macrophages and lymphocytes decreased with rIL-8 administration, suggesting that they were not attracted at the same rate as neutrophils. (C) 2007 Elsevier Ltd. All rights reserved.

Two cathelicidin genes are present in both rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar)

Further to the previous finding of the rainbow trout rtCATH_1 gene, this paper describes three more cathelicidin genes found in salmonids: two in Atlantic salmon, named asCATH_1 and asCATH_2, and one in rainbow trout, named rtCATH_2. All the three new salmonid cathelicidin genes share the common characteristics of mammalian cathelicidin genes, such as consisting of four exons and possessing a highly conserved preproregion and four invariant cysteines clustered in the C-terminal region of the cathelin-like domain. The asCATH_1 gene is homologous to the rainbow trout rtCATH_1 gene, in that it possesses three repeat motifs of TGGGGGTGGC in exon IV and two cysteine residues in the predicted mature peptide, while the asCATH_2 gene and rtCATH_2 gene are homologues of each other, with 96% nucleotide identity. Salmonid cathelicidins possess the same elastase-sensitive residue, threonine, as hagfish cathelicidins and the rabbit CAP18 molecule. The cleavage site of the four salmonid cathelicidins is within a conserved amino acid motif of QKIRTRR, which is at the beginning of the sequence encoded by exon W. Two 36-residue peptides corresponding to the core part of rtCATH_1 and rtCATH_2 were chemically synthesized and shown to exhibit potent antimicrobial activity. rtCATH_2 was expressed constitutively in gill, head kidney, intestine, skin and spleen, while the expression of rtCATH_1 was inducible in gill, head kidney, and spleen after bacterial challenge. Four cathelicidin genes have now been characterized in salmonids and two were identified in hagfish, confirming that cathelicidin genes evolved early and are likely present in all vertebrates.

Regulation of gamete release in the economic brown seaweed Hizikia fusiforme (Phaeophyta)

Gamete release is an essential event in artificial seeding of the economic brown seaweed, Hizikia fusiforme. Mass egg release occurred in the dark, with few eggs being discharged in the light. Release of eggs was elicited with eight practical salinity units (one PSU = 1 g sea salts l(-1)) and was inhibited by salinity levels > 32 PSU. Egg release was optimal at 23 degrees C, and was decreased by 72% in agitated seawater compared to unstirred seawater. Inhibitors of photosynthesis and ions channels suppressed egg release, indicating that this process was physiologically associated with photosynthetic activity and ion transport.

Identification of a novel cathelicidin gene in the rainbow trout, Oncorhynchus mykiss

We report the cloning of a novel antimicrobial peptide gene, termed rtCATH_1, found in the rainbow trout, Oncorhynchus mykiss. The predicted 216-residue rtCATH_1 prepropeptide consists of three domains: a 22-residue signal peptide, a 128-residue cathelin-like region containing two identifiable cathelicidin family signatures, and a predicted 66-residue C-terminal cationic antimicrobial peptide. This predicted mature peptide was unique in possessing features of different known (mammalian) cathelicidin subgroups, such as the cysteine-bridged family and the specific amino-acid-rich family. The rtCATH_1 gene comprises four exons, as seen in all known mammalian cathelicidin genes, and several transcription factor binding sites known to be of relevance to host defenses were identified in the 5' flanking region. By Northern blot analysis, the expression of rtCATH_1 was detected in gill, head kidney, and spleen of bacterially challenged fish. Primary cultures of head kidney leukocytes from rainbow trout stimulated with lipopolysaccharide or poly(I (.) C) also expressed riCATH_1. A 36-residue peptide corresponding to the core part of the fish cathelicidin was chemically synthesized and shown to exhibit potent antimicrobial activity and a low hemolytic effect. Thus, rtCATH_1 represents a novel antimicrobial peptide gene belonging to the cathelicidin family and may play an important role in the innate immunity of rainbow trout.