Natural Stable Isotopes for Determination of Gastrointestinal Transit Time in Fish

This study evaluated the application of stable isotopes of carbon as an alternative and more accurate method to determine gastrointestinal transit time (GTT) in fish by comparing it to the inert marker method. The stable isotope method detects alterations of the normal carbon flow in a biological system by analyzing naturally occurring isotopes of carbon, contrary to studies based on conventional techniques that apply external markers to the diet to determine GTT through visual observation of the color change in feces. Therefore, 320 pacu, Piaractus mesopotamicus juveniles were reared in 32 tanks under two different temperatures (25 and 29 C). The pacu juveniles received two different diets, one based on ingredients derived from C₃ photosynthetic cycle plants and the other based on C₄ plant ingredients, both containing titanium oxide (TiO₂) as a marker. After 40 d, the isotopic signature of the diets was changed, and the marker was replaced by chromic oxide (Cr₂O₃). In the isotopic technique, the feces were analyzed to determine the exchange in the isotopic ratio of carbon δ¹³C. Both methods found that GTT was faster (nearly 6 h) in fish at 29 C when using the C₄/C₃ feeding strategy and slower in fish at 25 C using the C₃/C₄ strategy (15 h by inert marker and 18 h by the isotopic method). In conclusion, GTT determination in pacu juveniles using the stable isotope technique exhibits the same accuracy obtained with the inert marker method at temperatures suitable (nearly 29 C) for the metabolism of these animals.     

Blue tilapia Oreochromis aureus (Steindachner) carcass composition and haematology in relation to the female parent size under recirculated water conditions

The relation between female parent size/age of Oreochromis aureus (Steindachner) and their offspring carcass composition and some haematological parameters was investigated. Blood parameters (glucose, triglycerides, albumin, cholesterol, total lipid, haematocrit, osmolarity), proximate carcass composition (crude protein, crude fat, moisture, ash), and total lipid content of liver were evaluated. No clear differences between populations could be reported regarding carcass composition. However, the offspring of the smallest/youngest female parent gave the highest crude protein and the lowest crude fat levels, while that of the largest/old female parent showed an opposite picture. Results obtained on blood parameters and liver lipid content were not clearly related to the female parent size/age, nor to the final body weight of the populations, despite the statistically significant (P < 0.05) differences observed for glucose, triglycerides and plasma and liver total lipids. It is concluded that more research is needed in order that a clear correlation between the female parent size/age and the investigated biochemical parameters of their progeny to be established.     

Inheritance of resistance to angular leaf spot in common bean and validation of the utility of resistance linked markers for marker assisted selection out side the mapping population

Inheritance of resistance to angular leaf spot (ALS) disease caused by Phaeoisariopsis griseola (Sacc.) Ferr was investigated in two common bean cultivars, Mexico 54 and BAT 332. Both Andean and Mesoamerican backgrounds were used to determine the stability of the resistance gene in each of the two cultivars. Resistance to P. griseola was phenotypically evaluated by artificial inoculation with one of the most widely distributed pathotypes, 63–39. Evaluation of the parental genotypes, F₁, F₂ and backcross populations revealed that the resistance to angular leaf spot in the cultivars Mexico 54 and BAT 332 to pathotype 63–39 is controlled by a single dominant gene, when both the Andean and Mesoamerican backgrounds were used. Allelism test showed that ALS resistance in Mexico 54 and BAT 332 to pathotype 63–39 was conditioned by the same resistance locus. Resistant and susceptible segregating populations generated using Mexico 54 resistant parent were selected for DNA extraction and amplification to check for the presence /absence of the SCAR OPN02 and RAPD OPE04 markers linked to the Phg-2 resistance gene. The results indicated that the SCAR OPN02 was not polymorphic in the study populations and therefore of limited application in selecting resistant genotypes in such populations. On the other hand, the RAPD OPE04 marker was observed in all resistant individuals and was absent in those scored susceptible based on virulence data. Use of the RAPD OPE04 marker in marker-assisted selection is underway.     

Comparison of the effects of six stunning/killing procedures on flesh quality of sea bass (Dicentrarchus labrax,Linnaeus 1758) and evaluation of clove oil anaesthesia followed by chilling on ice/water slurry for potential implementation in aquaculture

The effects of different stunning/killing procedures on flesh quality of European sea bass were investigated: (1) anaesthesia with clove oil, (2) anaesthesia with 2‐phenoxyethanol, (3) percussive stunning, (4) immersion in ice/water slurry, (5) chilling on ice and (6) anaesthesia with clove oil followed by immersion in ice/water slurry. Muscle pH values were significantly lower in sea bass anaesthetized or stunned by a blow to the head compared with fish immersed in ice/water slurry, chilled on ice or immersed in ice/water slurry after clove oil anaesthesia. Lightness was highest in sea bass anaesthetized by 2‐phenoxyethanol or percussively stunned and lowest in ice‐chilled fish. Redness and yellowness were highest in fish chilled on ice and lowest in fish anaesthetized with clove oil. Liquid loss, fat loss and shear values were not significantly different among the procedures. In general, lipid oxidation values during refrigerated or frozen storage did not significantly differ among treatment groups. Combination of clove oil anaesthesia followed by chilling on ice/water slurry appears to improve both flesh quality and welfare of sea bass, although the implementation of further studies is warranted to consolidate this finding.     

Antispasmodic effects of Persea cordata bark fractions on guinea pig ileum

The present study describes the antispasmodic activity of some fractions from the bark of Persea cordata, against guinea pig ileum contracted by acetylcholine and histamine. Hexane and dichloromethane fractions demonstrated the most promising biological effects, having similar potency to that of papaverine, a well-known antispasmodic agent.     

Secretion of multi-protein migratory complex induced by Toxoplasma gondii infection in macrophages involves the uPA/uPAR activation system

Toxoplasmosis is a world wide spread zoonosis caused by Toxoplasma gondii, an obligate intracellular parasite that is able to disseminate into deep tissues and cross biological barriers, reaching immunoprivileged sites such as the brain and retina. The parasite is able to infect macrophages and dendritic cells for dispersal throughout the body. However, the molecular mechanisms or outcomes of the subversion of the host cell are largely unknown. Recently our group established that metalloproteinases are involved in migration of infected macrophages. Herein, we evaluated the recruitment of host invasive machinery components in T. gondii infected murine macrophages. We showed by immunoprecipitation assays that MMP-9, CD44 TIMP-1 and uPAR were secreted as a multi-protein complex by infected macrophages. Zymographic analysis revealed that MMP-9 was present in its pro- and active form. Moreover, inhibition of uPA/uPAR pathway by PAI-1 decreased secretion of MMP-9 active forms, as well those associated to uPAR and TIMP-1, but not to CD44. Data presented here suggest that MMP-9 is secreted as a multiprotein complex by T. gondii infected macrophages, similar to that observed in metastatic cells. We further speculate that uPA/uPAR system is involved in the expression/secretion of complexes containing active MMP-9 forms.     

Nucleic acids and fatty acids of the common octopus, Octopus vulgaris, in relation to the growth rate

The aim of this study was to investigate changes in the ribonucleic acid (RNA)/deoxyribonucleic acid (DNA) ratio and the fatty acid composition of cultured Octopus vulgaris (50–750 g) in relation to recent (last 30 days) specific growth rate (SGR). Wild animals (80–500 g), collected in the field throughout the year (Aegean Sea, Mediterranean), were also examined for the aforementioned biochemical parameters. Octopuses were reared in a closed seawater system at three different temperatures (15, 20 and 25 °C). The octopuses were fed on squid (Loligo vulgaris). The cultured animals showed a high n‐3 (33–52.9%) and n‐6 (3.3–13.7%) polyunsaturated fatty acid content, but with a high variation for 22:6n‐3 [docosahexaenoic acid (DHA)], 20:5n‐3 [eicosapentaenoic acid (EPA)] and 20:4n‐6 [arachidonic acid (AA)]. The proportion of these fatty acids (% total fatty acids) and the RNA/DNA ratio were linearly (P<0.0001) related to SGR. Specifically, RNA/DNA (0.5–1.9) and AA (2.7–10.7%) increased, while EPA (10.4–19.7%) and DHA (20.8–31.9%) decreased, with increasing SGR (0.4–1.7% day⁻¹). The highest levels of SGR, RNA/DNA and AA were detected in small (50–150 g) octopuses reared at 20 and 25 °C and in large (500–750 g) animals reared at 15 °C. Similar RNA/DNA levels and fatty acid percentages were found in wild octopuses. It is concluded that RNA/DNA, DHA, EPA and AA could be used as biochemical indices for predicting the growth rate of O. vulgaris.     

Digestible protein requirements and muscle growth in juvenile tambaqui (Colossoma macropomum)

Several investigations have been carried out to improve the productivity of tambaqui, an economically important fish species in Brazil and other Latin American countries. This study determined the digestible protein (DP) requirements in juvenile tambaqui by assessing their productive performance and nutritional efficiency. It also evaluated the effects of different dietary DP levels on the morphology and cellularity of skeletal fast muscle fibres. The 1750 tambaqui tested (6.53 ± 0.43 g body weight, 7.58 ± 0.18 cm length) were randomly distributed into 35 tanks. Fish were fed one of the seven isocaloric diets, which contained 140, 170, 200, 230, 260, 290 or 320 g/kg DP. The DP requirement, calculated by segmented (broken line) regression of weight gain data, was 290 g/kg. An increase in diet DP to 290 g/kg significantly improved final weight, weight gain, feed intake, specific growth rate and crude protein gain, and changed fibre diameter in deep muscle. Muscle fibres were randomly distributed into a mosaic pattern, characterized by fibres with different diameters. Treatments with 290 and 320 g/kg DP increased the frequency of large‐diameter fibres (>50 µm), indicating hypertrophic growth of skeletal muscle during the juvenile phase, which occurred in conjunction with hyperplasia.     

Effects of background color on growth performances and physiological responses of scaled carp (Cyprinus carpio L.) reared in a closed circulated system

Growth performances and physiological responses of Cyprinus carpio after long-term (14 weeks) background color adaptation were investigated. Six groups of ten individuals each (initial body weight 116 g) were reared in black, green and white tanks (two replicate groups for each color). At the end of the experiment, blood (cortisol, glucose, haematocrit, cholesterol, triglycerides, total lipids, osmolality, electrolytes, pCO₂, pH), liver (total lipids, glycogen, hepatosomatic index) and growth (body weight, specific growth rate, food conversion ratio, condition factor, proximate carcass composition) parameters were determined. Plasma cortisol levels in white-adapted carp were significantly (P<0.05) lower than those in black, while in green-adapted fish did not differ significantly from those in both other counterparts. White-adapted carp showed the highest specific growth rate and the lowest food conversion ratio, whereas black-adapted fish exhibited the opposite pattern. In addition, mean (%) increase of body weight in white-adapted carp was 4.66 and 3.58% higher than that in black- and green-adapted fish, respectively. Furthermore in white-adapted carp, blood pCO₂ and pH were significantly higher and lower, respectively, than those obtained in black- and green-adapted fish. In black-adapted carp, liver total lipid levels were significantly lower, and plasma total lipid levels were significantly higher, than those in white- and green-adapted fish. No significant variations were observed in the other parameters. It is concluded that different background colors may lead to different growth performances of scaled carp depending upon rearing conditions.     

Fatty acid composition and biometry of five Greek Artemia populations suitable for aquaculture purposes

Evaluation of a new‐to‐science Artemia population and comparison with four other existing populations were carried out. Five parthenogenetic Artemia populations from the following Greek saltworks were studied: Alyki (a new population), Kalloni, Milos, Polychnitos and Messolongi. The diameters of non‐decapsulated and decapsulated cysts as well as the lengths of nauplii instar stages I, II and III were measured. In addition, the fatty acid composition of decapsulated cysts was estimated. The cyst size of the Alyki population was the smallest, with the largest chorion found in Greece so far. The Messolongi population had the largest cysts, with the smallest chorion of all five populations. The diameter of the non‐decapsulated cysts was highly related (P<0.01) to the length of the nauplii instar I, II and III. The diameter of decapsulated cysts was found to be the most stable and reliable biometric characteristic. The study of their fatty acids profiles revealed that the Polychnitos and Messolongi populations are suitable as live food for freshwater species, while the Alyki, Milos and Kalloni populations are suitable for marine species, having high levels (8.9%, 11.2% and 12.5% respectively) of eicosapentaenoic acid and detectable amounts (0.3%, 0.1% and 0.2% respectively) of docosahexaenoic acid.