Microbicidal effect of medicinal plant extracts (Psidium guajava Linn. and Carica papaya Linn.) upon bacteria isolated from fish muscle and known to induce diarrhea in children

Out of the twenty-four samples of shrimp and fish muscle used for this study, twelve were collected near a large marine sewer for waste disposal, 3 km off the coast of Fortaleza (Brazil) and used for the isolation of E. coli. Other twelve were collected at the Mucuripe fresh fish market (Fortaleza, Brazil) and used for the isolation of Staphylococcus aureus. Ethanol, water and acetone-diluted extracts of guava and papaya leaf sprouts were tested on the bacteria in order to verify their microbicidal potential. The E. coli strains used in the trials were rated LT positive. The papaya leaf extracts (Carica papaya Linn) showed no microbicidal activity while the guava sprout extracts (Psidium guajava Linn) displayed halos exceeding 13 mm for both species, an effect considered to be inhibitory by the method employed. Guava sprout extracts by 50% diluted ethanol most effectively inhibited E. coli (EPEC), while those in 50% acetone were less effective. It may be concluded that guava sprout extracts constitute a feasible treatment option for diarrhea caused by E. coli or by S. aureus-produced toxins, due to their quick curative action, easy availability in tropical countries and low cost to the consumer.

Application of Response Surface Methodology to study the effect of different calcium sources in fish muscle-alginate restructured products

Sodium alginate needs the presence of calcium ions to gelify. For this reason, the contribution of the calcium source in a fish muscle mince added by sodium alginate, makes gelification possible, resulting a restructured fish product. The three different calcium sources considered were: Calcium Chloride (CC); Calcium Caseinate (CCa); and Calcium lactate (CLa). Several physical properties were analyzed, including mechanical properties, colour and cooking loss. Response Surface Methodology (RSM) was used to determine the contribution of different calcium sources to a restructured fish muscle. The calcium source that modifies the system the most is CC. A combination of CC and sodium alginate weakened mechanical properties as reflected in the negative linear contribution of sodium alginate. Moreover, CC by itself increased lightness and cooking loss. The mechanical properties of restructured fish muscle elaborated were enhanced by using CCa and sodium alginate, as reflected in the negative linear contribution of sodium alginate. Also, CCa increased cooking loss. The role of CLa combined with sodium alginate was not so pronounced in the system discussed here.

Metallomic study of mercury in fish muscle (Semaprochilodus sp.) from Amazon - Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estimating Amino Acid Requirement of Brazilian Freshwater Fish from Muscle Amino Acid Profile

Information on nutritional requirement of some Brazilian farmed fish species, especially essential amino acids (EAA) requirements, is scarce. The estimation of amino acids requirements based on amino acid composition of fish is a fast and reliable alternative. Matrinxa, Brycon amazonicus, and curimbata, Prochilodus lineatus, are two important Brazilian fish with potential for aquaculture. The objective of the present study was to estimate amino acid requirements of these species and analyze similarities among amino acid composition of different fish species by cluster analysis. To estimate amino acid requirement, the following formula was used: amino acid requirement = [(amount of an individual amino acid in fish muscle tissue) x (average totalEAA requirement among channel catfish, Ictalurus punctatus, Nile tilapia, Oreochromis niloticus, and common carp, Cyprinus carpio)]/(average fish muscle totalEAA). Most values found lie within the range of requirements determined for other omnivorous fish species, in exception of leucine requirement estimated for both species, and arginine requirement estimated for matrinxa alone. Rather than writing off the need for regular dose-response assays under the ideal protein concept to determine EAA requirements of curimbata and matrinxa, results set solid base for the study of tropical species dietary amino acids requirements.

Comparison of the stable carbon and nitrogen isotopic values of gill and white muscle tissue of fish

The potential use of stable carbon and nitrogen isotope ratios (d13C, d15N) of fish gills for studies on fish feeding ecology was evaluated by comparing the d13C and d15N of gill tissue with the more commonly used white muscle tissue. To account for the effect of lipid content on the d13C signatures, a study-specific lipid correction model based on C:N ratios was developed and applied to the bulk d13C data. For the majority of species in the study, we found no significant difference in d13C values between gill and muscle tissue after correction, but several species showed a small (0.3-1.4 per mil) depletion in 13C in white muscle compared to gill tissue. The average species difference in d15N between muscle and gill tissue ranged from -0.2 to 1.6 per mil for the different fish species with muscle tissue generally more enriched in 15N. The d13C values of muscle and gill were strongly linearly correlated (R**2 = 0.85) over a large isotopic range (13 per mil), suggesting that both tissues can be used to determine long-term feeding or migratory habits of fish. Muscle and gill tissue bulk d15N values were also strongly positively correlated (R**2= 0.76) but with a small difference between muscle and gill tissue. This difference indicates that the bulk d15N of the two tissue types may be influenced by different isotopic turnover rates or a different composition of amino acids.

New approach for vitamin E extraction in rainbow trout flesh: Application in fish fed commercial and red seaweed-supplemented diets

A vitamin E extraction method for rainbow trout flesh was optimized, validated, and applied in fish fed commercial and Gracilaria vermiculophylla-supplemented diets. Five extraction methods were compared. Vitamers were analyzed by HPLC/DAD/fluorescence. A solid-liquid extraction with n-hexane, which showed the best performance, was optimized and validated. Among the eight vitamers, only α- and γ-tocopherol were detected in muscle samples. The final method showed good linearity (>0.999), intra- (<3.1%) and inter-day precision (<2.6%), and recoveries (>96%). Detection and quantification limits were 39.9 and 121.0 ng/g of muscle, for α-tocopherol, and 111.4 ng/g and 337.6 ng/g, for γ-tocopherol, respectively. Compared to the control group, the dietary inclusion of 5% G. vermiculophylla resulted in a slight reduction of lipids in muscle and, consequently, of α- and γ-tocopherol. Nevertheless, vitamin E profile in lipids was maintained. In general, the results may be explained by the lower vitamin E level in seaweed-containing diet. Practical Applications: Based on the validation results and the low solvent consumption, the developed method can be used to analyze vitamin E in rainbow trout. The results of this work are also a valuable information source for fish feed industries and aquaculture producers, which can focus on improving seaweed inclusion in feeds as a source of vitamin E in fish muscle and, therefore, take full advantage of all bioactive components with an important role in fish health and flesh quality.

Persistent organic pollutants in sediments and fish from Lake Victoria, Uganda

This thesis describes the occurrence and sources of selected persistent organic pollutants (POPs) such as polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs) and hexachlorocyclohexanes (HCHs) in the northern watershed of Lake Victoria. Sediments and fish were collected from three highly polluted embayments (i.e. Murchison Bay, Napoleon Gulf and Thurston Bay) of the lake. The analysis for PCDD/Fs, PCBs and PBDEs was done using a high resolution mass spectrometer coupled to a gas chromatograph (GC), and a GC equipped with an electron capture detector was used for HCHs. Total (Σ) PCDD/Fs, PCBs and PBDEs in sediments ranged from 3.19 to 478, 313 to 4325 and 60.8 to 179 pg g-1 dry weight (dw), respectively. The highest concentrations of pollutants were found at sites close to industrial areas and wastewater discharge points. The maximum concentrations of PCDD/Fs, PCBs, PBDEs and HCHs in fish muscle homogenates were 49, 779, 495 and 45,900 pg g-1 wet weight (ww), respectively. The concentrations of the pollutants in Nile perch (Lates niloticus) were significantly greater than those in Nile tilapia (Oreochromis niloticus), possibly due to differences in trophic level and dietary feeding habits among fish species. World Health Organization-toxic equivalency quotient (WHO2005-TEQ) values in the sediments were up to 4.24 pg g-1 dw for PCDD/Fs and 0.55 pg TEQ g-1 dw for the 12 dioxin-like PCBs (dl-PCBs). 23.1% of the samples from the Napoleon Gulf were above the interim sediment quality guideline value of 0.85 pg WHO-TEQ g-1 dw set by the Canadian Council for Ministers of the Environment. The WHO2005-TEQs in fish were 0.001-0.16 pg g-1 for PCDD/Fs and 0.001-0.31 pg g-1 ww for dl- PCBs. The TEQ values were within a permissible level of 3.5 pg g−1 ww recommended by the European Commission. Based on the Commission set TEQs and minimum risk level criteria formulated by the Agency for Toxic Substances and Disease Registry, the consumption of fish from Lake Victoria gives no indication of health risks associated to PCDD/Fs and PCBs. Principal component analysis (PCA) indicated that anthropogenic activities such as agricultural straw open burning, medical waste incinerators and municipal solid waste combustors were the major sources of PCDD/Fs in the watershed of Lake Victoria. The ratios of α-/γ-HCH varied from 0.89 to 1.68 suggesting that the highest HCH residues mainly came from earlier usage and fresh γ-HCH (lindane). In the present study, the concentration of POPs in fish were not significantly related to those in sediments, and the biota sediment accumulation factor (BSAF) concept was found to be a poor predictor of the bioavailability and bioaccumulation of environmental pollutants.

Raw-appearing Restructured fish models made with Sodium alginate or Microbial transglutaminase and effect of chilled storage

Restructuring by adding Sodium Alginate or Microbial Transglutaminase (MTGase) using cold gelation technology make it possible to obtain many different raw products from minced and/or chopped fish muscle that are suitable for being used as the basis of new restructured products with different physicochemical properties and even different compositions. Special consideration must be given to their shelf-life and the changes that may take place during chilling, both in visual appearance and physicochemical properties. After chilled storage, the restructured models made with different muscular particle size and composition at low temperature (5 °C), it was observed that microbial growth limited the shelf-life to 7-14 days. Mechanical properties increased (p < 0.05) during that time, and higher values were observed in samples elaborated by joining small muscle particle size than in those elaborated by homogenization. There was no clear increase in the cooking yield and purge loss, and no significant colour change (p > 0.05) was detected during storage.

Effect of previous chilling storage on quality loss in frozen (–20 °C) sierra (Scomberomorus sierra) muscle packed with a low-density polyethylene film containing butylated hydroxytoluene

Rancidity development during frozen storage (–20 °C) of sierra fish (Scomberomorus sierra) was studied. Fillets were packed in low-density polyethylene films with and without butylated hydroxytoluene added (BHT-LDPE and LDPE respectively). Fillets stored with no package were used as control. Special attention was given to the effect of previous ice storage (0, 3, 6, 9 and 15 days) on the quality of the frozen fish. Physical (pH and texture) and chemical (peroxide value, PV and thiobarbituric acid index, TBA-i) analyses were carried out. Lipid oxidation increased with ice storage time in fish muscle without film packing, being greater than the film packed muscle (with and without antioxidant). An effect of previous ice storage time was observed on the frozen product (in all treatments). However, fish muscle with film packing containing antioxidant showed less lipid deterioration. Under the conditions applied in this study, the plastic films with antioxidant prevented the lipids oxidation during the cold handling of the sierra muscle.

Enzyme reactions as index of freshness of Fish and Shellfish

An attempt has been made in this study to screen some fish muscle enzymes to assess their potential worth in testing the degree of freshness of fish. A problem with routine enzyme activity determinations is the complexity of the method of enzyme assay. Hence, in the present study as far as possible simple assay techniques were adopted. Several species were screened to assess the possibility of employing this procedure on a large scale. It is hoped that findings of this study will lead to the development of meaningful criteria in testing the freshness of fish. This thesis has been divided into five chapters

Muscle growth in Nile tilapia (Oreochromis niloticus): histochemical, ultrastructural and morphometric study

Muscle growth in Nile tilapia (Oreochromis niloticus) was studied focusing on histochemical, ultrastructural, and morphometric characteristics of muscle fibers. Based on body length (cm), we studied four groups: G1 = 1.36+/-0.09, G2 = 3.38+/-0.44, G3 = 8.90+/-1.47, and G4 = 28.30+/-3.29 (mean+/-S.D.). All groups showed intense reaction to NADH-TR in subdermal fibers and weak or no reaction in deep layer fibers. In G3 and G4, an intermediate layer was also observed with fibers presenting weak reaction; in G4, groups of fibers with intense reaction were observed in the subdermal region. The myosin ATPase (m-ATPase) activities were acid-stable and alkali-labile in subdermal fibers; most deep layer fibers were alkali-stable and acid-labile. Intermediate fibers were acid-labile and alkali-stable. Two fiber populations were observed near deep muscle layer: one large presenting weak acid- and alkali-stable and the other small alkali-stable.During growth, muscle fiber hypertrophy was more evident in intermediate and white fibers for G3 and G4. However, in these groups, the presence of fiber diameters less than or equal to21 mum suggested that there is still substantial fiber recruitment, confirmed by ultrastructural study, but hypertrophy is the main mechanism contributing to increase in muscular mass. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

Effects of addition of a bird repellent to fish diets on their growth and bioaccumulation

The effects of adding the nonlethal bird repellent methyl anthranilate (MA), at levels of 100 and 1000 mg/kg, to fish feed on the bioaccumulation and growth of juvenile (10 g) hybrid striped bass (Morone chrysops x M. saxatilis) and juvenile (1g) African cichlid fish Aulonocara jacobfreibergi were investigated under laboratory conditions. The bird repellent did not have any effect on the fish growth or survival over a period of 6 weeks. MA residues at low levels of 11.2 ± 2.6 mg/g were found in lipophilic tissues (liver) of MA-fed fish. Control fish, which had no MA added to their diet, had a much lower level of 0.6 ± 0.3 mg/g MA in their liver. Fish muscle was found to contain negligible MA residues, while the outer body surface mucus did not contain any MA. Following a 6-week depuration period, during which the previously MA-fed hybrid striped bass were fed a feed to which no MA was added, the levels of MA residues detected were reduced by one order of magnitude.

Nutritional composition, quality and spoilage capacity of orange roughy (Hoplostethus atlanticus) from the North East Atlantic

The nutritional composition o f orange roughy (collected from the Northeast Atlantic near the Rockall Trough) was studied on a seasonal basis. In addition samples were aged and stability assessed. Protein levels (16.68-16.21% w/w) were found to be slightly higher than those recorded for the N ew Zealand species o f orange roughy and compared favourably with protein values for fish muscle in general. Statistically results show a significant seasonal variation with no variation from fish to fish or in the location within the fish. Lipid content (3.6-4.5% w/w) was found to be much lower than that recorded for New Zealand. As with protein statistically results show a significant seasonal variation and no variation from fish to fish or in the location within the fish. Moisture levels (77.3_79.6%w/w) compared favourably with values obtained from other studies. Again statistically results show a significant seasonal variation with no variation from fish to fish or within the fish. Iodine values (74.63-79.54) indicate the likely presence o f a high level o f mono unsaturated fatty acids. Statistically results show no significant seasonal variation and no sample variation or variation within fish. Thin layer chromatography o f the extracted fat showed the major type to be wax esters with a much lower amount o f triglycerides and smaller amounts of polar lipids, free sterols and free fatty acids. Total fatty acid composition was found to be very similar to that recorded from other studies and showed that most o f the oils extracted from the fish muscle contained a high percentage o f mono unsaturates namely 16:1,18:1, 20:1 and 22:1 (85.63 - 91.14% ) with 16:1 present in the smallest amounts and 18:1 the major one. The only saturated fatty M.Sc. in Biochemistry III Nutritional Composition, Quality and Spoilage Capacity of Specific Deep Sea Fish acids present in significant quantities were 14:0, 16:0 and 18:0, the total varied from a seasonal average high o f 4.05 % to an average low o f 2.27%. The polyunsaturated fatty acids linoleic and arachidonic acid were present in small quantities varying in total from 0.89% to 1.50%. Docosapentaenoic acid (D P A ) was found only in trace quantities in spring, autumn and winter samples and undetected in summer. Levels o f Eicosapentaenoic acid (EPA ) and Docosahexaenoic acid (D H A ) were also found in very low percentages and varied on a seasonal basis with average values ranging from 0.41% in summer to 1.03 % in autumn for EPA and from 1.44 % in summer to3.20 % in autumn for D H A . Again statistically results show a significant seasonal variation with no variation from fish to fish or location within the fish. Levels o f freshness were measured using the Thiobarbituric acid (T B A ), Total volatile base nitrogen (T V B -N ) and Trimethylamine (T M A ) techniques. The quality o f the fish upon arrival was excellent and well below legal/acceptable lim its.T V B -N values ranged from 6.88-8.91 mg/lOOg and T M A values from 4.82-6.46 mg/lOOg Values for T B A ranged from 0.18-0.35 mg Malonaldehyde/kg fish. The summer values were higher than the other seasons. Seasonal variation was significant for all methods with no variation from fish to fish or within the fish. Fish aged at +4°C in air did not exceed the T V B N lim it o f 35mg/100g until day 6 whereas the T V B N lim it was extended to 8 days for fish aged at +4°C in vacuum. However the T M A lim it o f 12mg/100g was reached on day 4 for fish stored at +4°C in air and on day 5 for vacuum packed samples stored at +4°C . Fish stored at -5°C in air and vacuum packed did not reach the T V B N lim it until day 61 but the T M A limit was reached on day 24 for fish stored at -5°C in air and was extended to 31 days for vacuum packed fish stored at-5°C. Prolonged storage at -18°C caused some deterioration o f the frozen fish muscle. Upon thawing the shelf life o f fish stored for 12 months was much shorter than that stored for 6 M.Sc. in Biochemistry IV Nutritional Composition, Quality and Spoilage Capacity of Specific Deep Sea Fish months. This in turn deteriorated faster than fresh fish held at refridgeration temperature in air. Orange roughy were found to be a good source of protein with moisture levels similar to that o f other fish. They were o f medium fat content but have a very poor content o f the essential omega 3 and omega 6 fatty acids. Orange roughy can be stored at -18°C but its subsequent refridgerated shelf life will be shorter than that o f unfrozen orange roughy stored at refridgeration temperature. Orange roughy are a very important part o f the ecosystem. Their composition is less nutritionally beneficial than more readily available fish for human consumption and therefore should not be fished at all

Analysis of laboratory intercomparison data: a matter of independence

When laboratory intercomparison exercises are conducted, there is no a priori dependence of the concentration of a certain compound determined in one laboratory to that determined by another(s). The same applies when comparing different methodologies. A existing data set of total mercury readings in fish muscle samples involved in a Brazilian intercomparison exercise was used to show that correlation analysis is the most effective statistical tool in this kind of experiments. Problems associated with alternative analytical tools such as mean or paired 't'-test comparison and regression analysis are discussed.