Determination of moisture content and water activity in algae and fish by thermoanalytical techniques

The water content in seafoods is very important since it affects their sensorial quality, microbiological stability, physical characteristics and shelf life. In this study, thermoanalytical techniques were employed to develop a simple and accurate method to determine water content (moisture) by thermogravimetry (TG) and water activity from moisture content values and freezing point depression using differential scanning calorimetry (DSC). The precision of the results suggests that TG is a suitable technique to determine moisture content in biological samples. The average water content values for fish samples of Lutjanus synagris and Ocyurus chrysurus species were 76.4 ± 5.7% and 63.3 ± 3.9%, respectively, while that of Ulva lactuca marine algae species was 76.0 ± 4.4%. The method presented here was also successfully applied to determine water activity in two species of fish and six species of marine algae collected in the Atlantic coastal waters of Bahia, in Brazil. Water activity determined in fish samples ranged from 0.946 - 0.960 and was consistent with values reported in the literature, i.e., 0.9 - 1.0. The water activity values determined in marine algae samples lay within the interval of 0.974 - 0.979.

Evaluation of hard gelatin capsules and hydroxypropyl methylcellulose containing ampicillin

This study aims to develop and evaluate formulations containing ampicillin in capsules of gelatin and hydroxypropyl methylcellulose (HPMC). Two formulations (A and B) were developed. The final product quality was evaluated by testing for quality control and the results were in agreement with the Brazilian Pharmacopoeia. The formulations with HPMC capsules showed lower percentages of drug dissolved (99.67%, HPMC-A and 87.70%, HPMC-B) than the gelatin (100.18%, GEL-A and 101.16% GEL-B). Because of the delay of the ampicillin release observed in the dissolution profiles, it becomes necessary to evaluate the drugs that can be conditioned in the HPMC capsules.

Crystallographic studies of fish hemoglobins

The present work reports our succesfull experience concerning crystallization of four fish hemoglobins from three Brazilian species of Teleosts: Liposarcus anisitsi, Brycon cephalus and Piaractus mesopotamicus. The data shown here is part of a systematic functional and structural study of fish hemoglobins with the aim of better understanding the outstanding range of functional and structural properties exhibited by these proteins. We also present a reduced sparse-matrix method for crystallization of fish hemoglobins, which can reduce the amount of hemoglobin initially used in the crystallization experiments.

Production of a product similar to gelatin from chicken feet collagen

Chicken feet can be used as an alternative source of collagen for the development of new products. In this sense, the aim of this study was the production of a product similar to gelatin from collagen extracted from chicken feet and the evaluation of sensory quality. The products were produced in two distinct flavors, with grape flavor called GU and pineapple flavor called GA. Subsequently, we compared these formulations with gelatin of a trademark established in the market. We used in the verification of sensory acceptability of products a hedonic scale of 9 points and the availability of consuming the product by 30 untrained tasters. According to the results, all formulations showed good levels of acceptability, indicating the collagen from chicken feet as an alternative source of high quality in the production of gelatin.

Molecular characterization of virulence factors in Aeromonas hydrophila obtained from fish

Multiple factors can be involved in the virulence processes of Aeromonas hydrophila. The objective of the present paper was to verify the presence of aerolysin, hidrolipase, elastase and lipase virulence genes through the polymerase chain reaction (PCR) in A. hydrophila isolates obtained from fish of the São Francisco River Valley, and to evaluate virulence according to the presence of these genes in Nile tilapia fingerlings. One hundred and fourteen isolates from the bacteria were used. DNA was heat extracted and PCR undertaken using specific primers described in the literature. For in vivo tests Nile tilapia fingerlings were used. From the PCR tests, negative isolates for all genes tested were selected, positive isolates for two genes (aerolysin and elastase) and positive for the four genes tested. These were inoculated at a concentration of 10(8) UFC/ml into the tilapias, considered as treatments; another group of animals was used as control (with inoculation of saline solution). In all, 12 distinct standards regarding the presence of virulence factors in isolates from A. hydrophila, were observed. Of the 114 isolates analyzed, 100 (87.72%) presented at least one of the virulence factors under study. The virulence factors were widely distributed among the A. hydrophila isolates. Aerolysin was the most frequent virulence factor present in the isolates analyzed. A. hydrophila led to the mortality of the Nile tilapia fingerlings, regardless of the absence or quantity of virulence genes tested.

Parasites of the freshwater fish trade in Brazil: science metric study

This paper presents a science metric study of parasites of fish farming in Brazil, including a significant review of the literature. The methodology used was based on researching articles in three different databases, carried out ​on May 2012: ISI (Institute for Scientific Information), SciELO (Scientific Electronic Library Online), and Google Academic. The number of articles on fish parasites is mounting (currently over 110), having much increased since 1995. However, the quantity is still low compared with the amount of papers on parasites of fish from natural environments. In Brazil, the farmed fish that have been studied the most are pacu, tilapia and tambaqui. Monogeneans represent the most prevalent group, followed by protozoa and crustaceans. The regions most researched were the southeast and south, making up 84% of the total literature. The main issue addressed in articles was pathology, followed by treatment and record. In conclusion, the treatment of parasitic diseases of farmed fish in Brazil is still incipient, highlighting the importance and usefulness of management practices to prevent the occurrence of health problems.

Effects of the 2,4-D herbicide on gills epithelia and liver of the fish Poecilia vivipara

The 2,4-dichlorophenoxyacetic acid, usually named 2,4-D is one of the most widely used herbicides in the world. Acute toxicity of 2,4-D herbicide was investigated through its effects on guppies (Poecilia vivipara Bloch et Schneider 1801). Fish were exposed to the herbicide at concentrations of 10, 20 and 40µl per liter of water for 24 hours to determine its effects on gills and liver epithelia. The estimated LC50 was 34.64µl of 2,4-D per liter of water. Histochemical analyses and Feulgen's reaction were conducted to detect glycoconjugates and DNA, respectively, in gills and liver epithelia. Histochemistry revealed qualitative variations of glycoconjugates present on mucous cells and granules. The four types of mucous cells contained neutral granules, acids, or both. Increasing amounts of syalomucins were observed from the control group to the group exposed to the highest concentration of 2,4-D, suggesting increased mucous viscosity and the formation of plaques that could inhibit gas exchange and osmoregulation. Lamellar fusion observed in the group exposed to 40µl of 2,4-D suggests a defense mechanism. Hepatocytes showed vacuolization in the 10 and 20µl/L groups. The 40 µl/L group showed normal hepatocytes as well as changed ones, many Ito cells, micronuclei, and nuclear swelling. These effects may be associated with toxicity or adaptative processes to cellular stress. The data from this study indicates the importance of assessing similar risks to aquatic species and suggests that Poecilia vivipara is an adequate biological model for analysis of environmental contamination.

First description of microchromosome in the Anostomidae fish Schizodon nasutus from Argentina

Thirty-six specimens of Schizodon nasutus (Anostomidae-Characiformes) from the middle Paraná River (Posadas, Argentina) were analyzed cytogenetically. The karyotype of this species was similar to those described for this species in the literature. C-banding technique showed a rich heterochromatic pattern relative to other Anostomidae species. The NORs were located on one chromosome pair in terminal position and showed a very marked size heteromorphism. A microchromosome was observed with a frequency of about 20% in the sample studied. This additional element was punctiform, negative C-band, and constant in all metaphase plates of the seven carriers. The present study is the first karyotypic approach to Schizodon nasutus from Argentina and the first description of microchromosome in Anostomidae

Optimized Fast-FISH with a-satellite probes: acceleration by microwave activation

It has been shown for several DNA probes that the recently introduced Fast-FISH (fluorescence in situ hybridization) technique is well suited for quantitative microscopy. For highly repetitive DNA probes the hybridization (renaturation) time and the number of subsequent washing steps were reduced considerably by omitting denaturing chemical agents (e.g., formamide). The appropriate hybridization temperature and time allow a clear discrimination between major and minor binding sites by quantitative fluorescence microscopy. The well-defined physical conditions for hybridization permit automatization of the procedure, e.g., by a programmable thermal cycler. Here, we present optimized conditions for a commercially available X-specific a-satellite probe. Highly fluorescent major binding sites were obtained for 74oC hybridization temperature and 60 min hybridization time. They were clearly discriminated from some low fluorescent minor binding sites on metaphase chromosomes as well as in interphase cell nuclei. On average, a total of 3.43 ± 1.59 binding sites were measured in metaphase spreads, and 2.69 ± 1.00 in interphase nuclei. Microwave activation for denaturation and hybridization was tested to accelerate the procedure. The slides with the target material and the hybridization buffer were placed in a standard microwave oven. After denaturation for 20 s at 900 W, hybridization was performed for 4 min at 90 W. The suitability of a microwave oven for Fast-FISH was confirmed by the application to a chromosome 1-specific a-satellite probe. In this case, denaturation was performed at 630 W for 60 s and hybridization at 90 W for 5 min. In all cases, the results were analyzed quantitatively and compared to the results obtained by Fast-FISH. The major binding sites were clearly discriminated by their brightness

Electrolocation-communication discharges of the fish Gymnotus carapo L. (Gymnotidae: Gymnotiformes) during behavioral sleep

Technical problems have hampered the study of sleep in teleosts. The electrical discharges of Gymnotus carapo L. (Gymnotidae: Gymnotiformes) were monitored to evaluate their ease and reliability as parameters to study sleep. The discharges were detected by electrodes immersed in a glass aquarium and were recorded on a conventional polygraph. G. carapo showed conspicuous signs of behavioral sleep. During these periods, opercular beat rates were counted, electric discharges recorded, and the "sharp discharge increase" (SDI) of the orienting reflex was investigated. All 20 animals monitored maintained electrical discharges during behavioral sleep. The discharge frequencies during sleep (50.3 ± 10.4 Hz) were not significantly different from those observed when the fish was awake and inactive (57.2 ± 12.1 Hz) (Wilcoxon matched-pairs signed-ranks test, P>0.05). However, the SDI, which was prevalent in the awake fish, was not observed during periods of behavioral sleep. Additional observations showed that the species had cannibalistic habits. When presented with electrical discharges from a conspecific, the sleeping fish showed an initial decrease or pause in discharge frequency, while the awake fish did not have this response. We conclude that the electrical discharges of G. carapo were not conspicuous indicators of behavioral sleep. Discharges may have been maintained during sleep for sensory purposes, i.e., conspecific detection and avoidance of cannibalistic attacks.

Influence of season and pollution on the antioxidant defenses of the cichlid fish acará (Geophagus brasiliensis)

The livers of Geophagus brasiliensis collected from both a non-polluted site and a polluted site were analyzed for different antioxidant defenses, O2 consumption, thiobarbituric acid-reactive substance (TBARS) levels, and histological damage. Compared to controls (116.6 ± 26.1 nmol g-1), TBARS levels were enhanced at the polluted site (284.2 ± 25.6 nmol g-1), as also was oxygen consumption (86.6 ± 11.3 and 128.5 ± 9.8 µmol O2 min-1 g-1, respectively). With respect to enzymatic antioxidants, increased catalase activities (8.7 ± 1.3 and 29.2 ± 2.4 mmol min-1 g-1, respectively), unchanged superoxide dismutase activities (767.2 ± 113.3 and 563.3 ± 70.2 U g-1, respectively), and diminished glutathione S-transferase activities (29.0 ± 3.2 and 14.9 ± 3.2 µmol min-1 g-1, respectively) were detected. Reduced glutathione (1.91 ± 0.17 and 1.37 ± 0.25 mM, respectively), oxidized glutathione (1.50 ± 0.20 and 0.73 ± 0.17 mM, respectively), and total glutathione (3.40 ± 0.26 and 2.07 ± 0.27 mM, respectively) concentrations were also below control values at the polluted site. Nevertheless, the observed ethoxyresorufin-O-deethylase activities (1.34 ± 0.11 and 16.7 ± 0.21 pmol min-1 mg-1, respectively) showed enhanced values at the polluted site. The main histological damage observed in the hepatocytes from fish collected at the polluted site was characterized by heavy lipid infiltration. Fish collected at the end of spring showed higher O2 consumption, higher superoxide dismutase and glutathione S-transferase activities, and higher total and oxidized glutathione concentrations compared to the beginning of autumn. No seasonal changes were observed in catalase activities, glutathione or TBARS levels. Fish chronically exposed to relatively high pollution levels seem to be unable to set up adequate antioxidant defenses, probably due to severe injury to their hepatocytes. The higher antioxidant defenses found at the end of spring are probably related to the enhanced activities during high temperature periods in thermoconforming organisms.

Environmental blue light prevents stress in the fish Nile tilapia

The present study aimed to test the effects of blue, green or white light on the stress response of the Nile tilapia, Oreochromis niloticus (L.). Each color was tested on two groups of isolated adult Nile tilapia (8 replicates each): one being subjected to confinement stress, and the other not (control). A different environmental color was imposed on each compartment by covering the light source with cellophane of the respective color (green or blue; no cellophane was used for white light). The intensity of green, white and blue lights was 250, 590 and 250 lux, respectively. Basal plasma cortisol levels were determined for each fish prior to the experimental procedures. The fish were confined by being displaced toward one side of the aquarium using an opaque partition for 1 h both in the morning and the afternoon of the two consecutive days of the test. At the end of this 48-h period, plasma cortisol levels were measured again. Basal cortisol levels (ng/ml) were similar for each group (ANOVA, F(2;42) = 0.77, P = 0.47). Thus, plasma cortisol levels were analyzed in terms of variation from their respective basal level. After confinement, plasma cortisol levels were not increased in fish submitted to a blue light environment. Thus, blue light prevents the confinement-induced cortisol response, an effect not necessarily related to light intensity.

Different biochemical strategies of two Neotropical fish to cope with the impairment of nitrogen excretion during air exposure

The exposure of fish to air is normally expected to interfere with the nitrogen excretion process. Hoplias malabaricus and Hoplerythrinus unitaeniatus, two teleost species, display distinct behaviors in response to decreases in natural reservoir water levels, although they may employ similar biochemical strategies. To investigate this point, plasma levels of ammonia, urea, uric acid, and the two urea cycle enzymes, ornithine carbamoyl transferase (OCT) and arginase (ARG), as well as glutamine synthetase (GS) were determined for both species after exposure to air. Plasma ammonia increased gradually during exposure to air, but only H. malabaricus showed increased concentrations of urea. Plasma uric acid remained very low in both fish. Enzymatic activities (mean ± SD, µmol min-1 g protein-1) of H. malabaricus showed significant increases (P<0.05, N = 6) in OCT from 0.84 ± 0.05 to 1.42 ± 0.03, in ARG from 8.07 ± 0.47 to 9.97 ± 0.53 and in GS from 1.15 ± 0.03 to 2.39 ± 0.04. The OCT and ARG enzymes remained constant in H. unitaeniatus (N = 6), but GS increased from 1.49 ± 0.02 to 2.06 ± 0.03. Although these species are very closely related and share the same environment, their biochemical strategies in response to exposure to air or to increased plasma ammonia are different.

Eye color as an indicator of social rank in the fish Nile tilapia

We investigated the association of eye color with the dominant-subordinate relationship in the fish Nile tilapia, Oreochromis niloticus. Eye color pattern was also examined in relation to the intensity of attacks. We paired 20 size-matched fish (intruder: 73.69 ± 11.49 g; resident: 75.42 ± 8.83 g) and evaluated eye color and fights. These fish were isolated in individual aquaria for 10 days and then their eye color was measured 5 min before pairing (basal values). Twenty minutes after pairing, eye color and fights were quantified for 10 min. Clear establishment of social hierarchy was observed in 7 of 10 pairs of fish. Number of attacks ranged from 1 to 168 among pairs. The quartile was calculated for these data and the pairs were then divided into two classes: low-attack (1 to 111 attacks - 2 lower quartiles) or high-attack (112 to 168 attacks - 2 higher quartiles). Dominance decreased the eye-darkening patterns of the fish after pairing, while subordinance increased darkening compared to dominance. Subordinate fish in low-attack confrontations presented a darker eye compared to dominant fish and to the basal condition. We also observed a paler eye pattern in dominants that shared low-attack interactions after pairing compared to the subordinates and within the group. However, we found no differences in the darkening pattern between dominants and subordinates from the high-attack groups. We conclude that eye color is associated with social rank in this species. Moreover, the association between eye color and social rank in the low-attack pairs may function to reduce aggression.

Effect of chronic fish oil supplementation on renal function of normal and cachectic rats

In the present study we determined the effect of chronic diet supplementation with n-3 PUFA on renal function of healthy and cachectic subjects by providing fish oil (1 g/kg body weight) to female rats throughout pregnancy and lactation and then to their offspring post-weaning and examined its effect on renal function parameters during their adulthood. The animals were divided into four groups of 5-10 rats in each group: control, control supplemented with fish oil (P), cachectic Walker 256 tumor-bearing (W), and W supplemented with fish oil (WP). Food intake was significantly lower in the W group compared to control (12.66 ± 4.24 vs 25.30 ± 1.07 g/day). Treatment with fish oil significantly reversed this reduction (22.70 ± 2.94 g/day). Tumor growth rate was markedly reduced in the P group (16.41 ± 2.09 for WP vs 24.06 ± 2.64 g for W). WP group showed a significant increase in mean glomerular filtration rate compared to P and control (1.520 ± 0.214 ml min-1 kg body weight-1; P < 0.05). Tumor-bearing groups had low urine osmolality compared to control rats. The fractional sodium excretion decreased in the W group compared to control (0.43 ± 0.16 vs 2.99 ± 0.87%; P < 0.05), and partially recovered in the WP group (0.90 ± 0.20%). In summary, the chronic supplementation with fish oil used in this study increased the amount of fat in the diet by only 0.1%, but caused remarkable changes in tumor growth rate and cachexia, also showing a renoprotective function.