Low Temperature Tolerance of Pacu, Piaractus mesopotamicus

Pacu, Piaractus mesopotamicus (=Colossoma mitrei), is a South American warm water fish species found in the temperature range of 15–35°C. The culture of a warm water species in temperate regions demands knowledge on its temperature requirements. Pacu introduction into the Israeli fish culture system is being considered. Temperature range in the region is 8–33°C, thus the minimum winter water temperatures might be a limiting factor. To determine what is the minimum temperature pacu would tolerate, and hence which overwintering operations in warm-temperate regions are required for this warm water species, low temperature tolerance tests in the laboratory and observations in the field were carried out. Laboratory experiments reducing temperature by 1–3°C per day were carried out with fish of 150–200g, about the size pacu reach after one culture season. The field observations compared survival of two-year-old pacu of 1.3kg mean weight overwintered in outdoors and in greenhouse ponds. For one-year-old fish 7.5°C was found to be the lower temperature tolerance limit. Two-year-old fish withstand short exposures to this temperature rather well and their lower tolerance limit might be lower. This indicates that in warm-temperate regions pacu should survive in outdoors ponds. In this case some loss of weight should be expected, and suspension of feeding when temperature drops below 16–18°C is recommended to avoid wasting feed that the fish will not consume anyway. To be in the safe side, inflow of the warmest available water into the ponds is recommended if maximum water temperature drops to 10°C or below. Overwintering in greenhouses or other heated facilities would be recommended if an exceptionally cold winter is expected and for regions with lower winter minimum temperatures.     

Temperature and hypoxia on oxidative stress responses in pacu Piaractus mesopotamicus

The aim of this study was to verify the effects of the interaction between different temperatures and levels of dissolved oxygen in the oxidative stress parameters of pacu juveniles. A total of 81 pacu juveniles (61.7 ± 9.1 g) were exposed to three temperatures (18, 23, and 28 °C), acclimated for a period of 30 days, and then submitted to three levels of dissolved oxygen: control or normoxia (7 mg L⁻¹); moderate hypoxia (4 mg L⁻¹); and severe hypoxia (2 mg L⁻¹) for 12 h. Glutathione-S-transferase (GST) activity, total antioxidant capacity against peroxyl radicals (ACAP), and protein thiol content (PSH) and LPO (lipid peroxidation) [measured by the TBARS] were measured in gill, liver, muscle and brain. The results indicated that the interaction between different temperatures and dissolved oxygen levels caused alterations in the antioxidant system and induced lipid and protein damage in pacu juveniles. In addition, the effects were organ specific. In conclusion, exposure to moderate and severe hypoxia affect oxidative stress parameters and have been shown to be organ-specific in pacu juveniles. The interaction between 23 °C and hypoxia caused greater disturbances in oxidative stress markers, such as PSH in the gills and liver and LPO in the muscle.     

Respiratory gill surface of the serrasalmid fish, Piaractus mesopotamicus

Gill element dimensions of pacu Piaractus mesopotamicus were estimated and correlated to body mass ( W ), according to the power equation Y=aW^b . The filament number ( b =0.154) and length ( b =0.457) increased with body mass, markedly influencing the respiratory gill surface area ( b =0.769). The high filament number and length, associated with a high secondary lamellae frequency ( a =40.21), are typical of active fish species and may be an adaptation to its migratory movements during reproduction. The comparatively small dimensions of its secondary lamellae are found more commonly in less active species, and may be related to the environmental conditions prevailing in lentic environments, where the species is normally found most of the year. Such features, together with its ability to compensate for oxygen reduction by means of a high ventilatory volume, and the use of aquatic surface respiration (ASR), may account for its adaptative capacity to withstand hypoxic conditions, with a low respiratory energy cost.     

Monoamine oxidase activity in kidney and heart of Piaractus mesopotamicus (Holmberg)

The values of Michaelis–Menten constant (K_M) and maximum velocity (V_MAX) for kidney and heart monoamine oxidase (MAO) from pacu Piaractus mesopotamicus were determined. The mean ± s . e . K_M values were 17·28 ± 2·27 μM for kidney and 15·38 ± 1·86 μM for heart. MAO activities were 111·60 ± 3·25 and 15·12 ± 0·30 nmols min⁻¹ g⁻¹ of wet tissue for kidney and heart, respectively. In addition, MAO inhibitory studies in these two tissues indicate that this enzyme may be a different isoform of MAO.     

Differential microRNA Expression in Fast- and Slow-Twitch Skeletal Muscle of Piaractus mesopotamicus during Growth

Pacu (Piaractus mesopotamicus) is a Brazilian fish with a high economic value in pisciculture due to its rusticity and fast growth. Postnatal growth of skeletal muscle in fish occurs by hyperplasia and/or hypertrophy, processes that are dependent on the proliferation and differentiation of myoblasts. A class of small noncoding RNAs, known as microRNAs (miRNAs), represses the expression of target mRNAs, and many studies have demonstrated that miR-1, miR-133, miR-206 and miR-499 regulate different processes in skeletal muscle through the mRNA silencing of hdac4 (histone deacetylase 4), srf (serum response factor), pax7 (paired box 7) and sox6 ((sex determining region Y)-box 6), respectively. The aim of our work was to evaluate the expression of these miRNAs and their putative target mRNAs in fast- and slow-twitch skeletal muscle of pacu during growth. We used pacus in three different development stages: larval (aged 30 days), juvenile (aged 90 days and 150 days) and adult (aged 2 years). To complement our study, we also performed a pacu myoblast cell culture, which allowed us to investigate miRNA expression in the progression from myoblast proliferation to differentiation. Our results revealed an inverse correlation between the expression of the miRNAs and their target mRNAs, and there was evidence that miR-1 and miR-206 may regulate the differentiation of myoblasts, whereas miR-133 may regulate the proliferation of these cells. miR-499 was highly expressed in slow-twitch muscle, which suggests its involvement in the specification of the slow phenotype in muscle fibers. The expression of these miRNAs exhibited variations between different development stages and between distinct muscle twitch phenotypes. This work provides the first identification of miRNA expression profiles in pacu skeletal muscle and suggests an important role of these molecules in muscle growth and in the maintenance of the muscle phenotype.     

Structural and morphological features of Piaractus mesopotamicus (Holmberg, 1887) gills

The pacu Piaractus mesopotamicus (Holmberg, 1887), is a commercially important serrasalmid fish endemic to the Paraná-Paraguay River basin, and one of the most widely cultivated native fish species in the Neotropics. As a migratory species, also inhabiting temporary pools subjected to hypoxic conditions, the species presents a high plasticity concerning respiratory adaptations. In order to supplement basic knowledge on the respiratory physiology and morphology of this species, some structural features of pacu gills, such as filament and lamellae structure, and circulatory pattern are described in this paper.     

Migratory movements of pacu, Piaractus mesopotamicus, in the highly impounded Paraná River

A mark‐recapture study was conducted in 1997–2005 to investigate movements of stocked pacu, Piaractus mesopotamicus, in the Paraná River Basin of Brazil, Paraguay, and Argentina. Fish raised in cages within the Itaipu Reservoir and in ponds were tagged externally (n = 2976) and released in the Itaipu Reservoir (53.2%) and bays of its major tributaries (46.8%). In total, 367 fish (12.3%) were recaptured. In all, 91% of the pacu moved away from the release site; upstream movements were more extensive than downstream movements. Pacu traveled upstream a maximum of 422 km (average of 41.3 km) at a maximum rate of 26.4 km day⁻¹ (av. 0.8). Downstream movements were limited in terms of number of individuals and distance moved. Fish released during the wet season moved farther than those released during the dry season, and feeding rather than spawning might have been the compelling reason for movement. Although fish passed downstream through dams, none of the marked fish were detected to have moved upstream through the passage facilities. Pacu showed movement patterns not radically different from those of other neotropical migratory species, but their migratory movements may not be as extensive as those of other large migratory species in the basin.     

Isolation and characterization of microsatellite loci in Piaractus mesopotamicus and their applicability in other Serrasalminae fish

The subfamily Serrasalminae (Characidae, Teleostei) is an endemic Neotropical group of fishes distributed along the Amazon, Paraná‐Paraguay and Orinoco Basins. The pacu (Piaractus mesopotamicus) is found in all major rivers that comprise the Pantanal of Mato Grosso. In order to investigate population genetic structure within the group, an enriched microsatellite library was constructed. Eight polymorphic were loci screened, with an average of 6.5 alleles per locus. Five loci exhibited greater than 60% heterozygosity. Additionally, a high level of cross‐species amplification was obtained.     

The administration of exogenous prostaglandin may improve ovulation in pacu (Piaractus mesopotamicus)

Based on the reports of unsuccessful ovulation in pacu (Piaractus mesopotamicus) by fish farmers and researchers undertaking artificial reproduction programs, we evaluated the use of prostaglandin F (PGF) to improve pacu ovulation. This study was conducted during two spawning seasons (2009/2010 and 2010/2011) with two samplings in the first season and one sampling in the second season. A total of 45 females was sampled in this study. The control group was injected with carp pituitary extract (crude extract, 6 mg/kg), and the treatment group received PGF (2 mL per fish in the 2009/2010 season and 5 mL per fish in the 2010/2011 season) in addition to the crude extract. In both seasons, 100% (N = 4, 2009/2010 first sampling; N = 5, 2009/2010 second sampling; and N = 3, 2010/2011) of the PGF-treated fish spawned. In contrast, 53.0% (N = 9) and 83.3% (N = 10) of the control fish spawned in the first and second samplings of the 2009/2010 season, respectively, and only 25.0% (N = 1) spawned in the 2010/2011 season. Fecundity, fertility, and hatching rates did not differ (P > 0.05) between the treated and control fish. Based on oocyte volume frequency analysis, ovaries of the control fish had more (P < 0.05) vitellogenic oocytes with germinal vesicle breakdown that remained unovulated after spawning, whereas more (P < 0.05) of previtellogenic oocytes were present in the ovaries of the PGF-treated fish. In conclusion, administration of exogenous prostaglandin may improve the outcome of hormonally induced spawning in tropical migratory fish.     

Trophic relationship between the Patinga hybrid (Piaractus mesopotamicus x Piaractus brachypomus) and Echinorhynchus gomesi Machado Filho, 1948 in fish farms

A total of 122 Patinga specimens were collected from fish farms (P1, P2 and P3), and only those from fish farm P1 were shown to be infected with Echinorhynchus gomesi. In addition, fish in this study were shown to have diets that consisted of 21 different food items, and Notodiaptomus sp. (Copepoda: Calanoida) was identified as a potential intermediate host for E. gomesi.     

Disturbance of pro-inflammatory cytokines in post-traumatic psychopathology

BACKGROUND: Post-traumatic psychopathology (PTP) is important to the orthopaedic surgeon as it may complicate the recovery from musculoskeletal injury. PTP is associated with a disturbance of the hypothalamic-pituitary-adrenal axis, and may lead to impaired healing. We have investigated the relationship between PTP and pro-inflammatory markers of the metabolic response to trauma. mATERIALS AND METHODS: A prospective cohort study of 82 patients with musculoskeletal injuries, correlating development of psychopathology (measured by general health questionnaire) and pro-inflammatory markers (CRP, IL-6, sIL-6r, TNF-alpha) two and six months after their injury. RESULTS: Psychological disturbance was found in 39% of patients at two months and 18% at six months. This disturbance was associated with significantly increased levels of IL-6 at two months and of sIL-6r and TNF-alpha at six months. CRP levels were not related to the development of PTP. CONCLUSIONS: The relationship between PTP and disturbances of pro-inflammatory markers needs further exploration, but may explain in part the impaired functional recovery when musculoskeletal trauma is complicated by psychological disturbance.     

Citric acid as a feed additive in pacu Piaractus mesopotamicus (Holmberg, 1887) diets

The objective of this study was to evaluate citric acid (0.0, 1.0, 2.0 and 3.0%) in isonitrogenous (23% of digestible protein) and isoenergetic (13.38 MJ of digestible energy/kg) pacu Piaractus mesopotamicus (Holmberg, 1887) diets. A 90‐day feeding trial was conducted to evaluate the growth performance, haematological parameters and pH of the diets, stomach and gut, somatic indices, nitrogen retention and body composition of pacu juveniles. Fish (n = 160, 12.53 ± 0.17g) were distributed in 16 aquaria (300‐L) with a recirculating water system (4 L/min) and controlled temperature (25.26 ± 0.47°C) in an experimental design completely randomized with four treatments and four replicates. Posteriorly, apparent digestibility coefficients (ADC) were assessed with pacu (n = 96, 80.35 ± 5.12 g) fed experimental diets including 0.1% chromium oxide III. Diet pH decreased (p < .05) with graded levels of citric acid to reduce pH in the stomach and gut. Pacu fed with 2.0% citric acid showed superior (p < .05) final weight at 30 days, compared to control; however, this did not differ by 60 and 90 days where was no difference (p > .05) in the haematology, somatic indices, body composition, or digestibility among treatments. The data showed that dietary citric acid improved the growth of pacu at 30 days, but had no long‐term effects on the digestibility of nutrients or the availability of P or Ca in the experimental diets.     

The effects of temperature and hyperoxia on arterial PO2 and acid-base status in Piaractus mesopotamicus

The effects of hyperoxia and change of temperature (range 20–30° C) on blood gases were studied in the teleost fish Piaractus mesopotamicus , native to several major river systems in Brazil. Large hyperoxia-induced increases of arterial P o₂ ( P _ao₂) indicated that true branchial blood shunts are negligible in relation to total gill perfusion. This implies that blood gases will be influenced by ventilation rather than by shunts. Acute variations of temperature ( t ) were accompanied by changes of arterial blood pH (on the average Δ p H_aΔt⁻¹ of — 0·015 units °C⁻¹), due mainly to alterations of P _aco₂: 2·4 mmHg at 20° C, 5·0 mmHg at 30° C. Concomitantly, P _ao₂ declined from 116 mmHg (20° C) to 89 mmHg (30° C). The data suggest that temperature-induced changes of acid-base status depend mainly on gill ventilation and that the decrease of P _ao₂ with higher temperature is a result of this regulation.     

Interactions between rnacrophage cytokines and eicosanoids in expression of antitumour activity

Cytokines and eicosanoid products of macrophages play an essential role in expression of antitumour activity of macrophages either in a cell-to-cell contact system between the effector and the target cell or as cell-free soluble products. In this review the relationship between three main monokines, namely TNF-alpha, IL-1 and IL-6 and the interrelationship between these monokines and eicosanoids (PGE(2), PGI(2), LTB(4), LTC(4)) in their production and in expression of antitumour activity is discussed. Emphasis is given to the effect of tumour burden on production of the monokines and of the eicosanoids and on the production of these compounds by the tumour cells. Finally, the therapeutic implications drawn from animal studies and clinical trials is discussed.     

Cytosolic glutathione peroxidase from liver of pacu (Piaractus mesopotamicus), a hypoxia-tolerant fish of the Pantanal

Pacu (Piaractus mesopotamicus Holmberg, 1887, Characiformes) dwells in waters of Pantanal, in which it has adapted for alternate concentrations of dissolved oxygen. Intracellular antioxidant protection should be vital for such an adaptation. Accordingly, we found that cytosol from liver of pacu has the highest antioxidant glutathione peroxidase activity so far reported for fish and murine species. To clarify whether this activity was due to a selenium independent glutathione S-transferase or to a glutathione peroxidase, we purified it and studied its kinetics. The substrates cumene hydroperoxide and hydrogen peroxide were promptly reduced by the enzyme, but peroxidized phosphatidylcholine had to undergo previous fatty acid removal with phospholipase A(2). Augmenting concentrations (from 2 to 6 mM) of reduced glutathione activated the pure enzyme. Curves of velocity versus different micromolar concentrations of hydrogen peroxide in the presence of 2, 4 or 8 mM reduced glutathione indicated that at least 2.5 mM reduced glutathione should be available in vivo for an efficient continuous destruction of micromolar concentrations of hydrogen peroxide by this peroxidase. Molecular exclusion HPLC and SDS-polyacrylamide gel electrophoresis indicated that the purified peroxidase is a homotetramer. Data from internal sequences showed selenocysteine in its primary structure and that the enzyme was a homologue of the type-1 glutathione peroxidase found in rat, bull, trout, flounder and zebra fish. Altogether, our data establish that in liver cells of pacu, a hypoxia-tolerant fish from South America, there are high levels of a cytosolic GPX-1 capable of quenching hydrogen peroxide and fatty acid peroxides, providing an effective antioxidant action.     

Antioxidant defenses and biochemical changes in the neotropical fish pacu, Piaractus mesopotamicus: Responses to single and combined copper and hypercarbia exposure

This study investigated the potentially detrimental effects of copper and elevated aquatic CO(2) (hypercarbia), alone or in combination, on pacu, Piaractus mesopotamicus. Fish were exposed for 48h to control (no copper addition in normocarbia), to 400μg Cu(2+)L(-1), to hypercarbic (1% CO(2); PCO(2)=6.9mm Hg) water and to 400μg Cu(2+)L(-1)+hypercarbia. In liver the single factors caused an increase in lipid hydroperoxide concentration that was not observed when the factors were combined. Copper exposure elicited increased hepatic superoxide dismutase activity, irrespective of aquatic CO(2) level. On the other hand, the effects of copper on hepatic glutathione peroxidase activity were dependent on water CO(2) levels. The two stressors combined did not affect hepatic catalase activity. Hypercarbic water caused a decline in plasma glucose concentration, but this was not observed when hypercarbia was combined with copper exposure. Copper caused a decrease in branchial Na(+)/K(+)-ATPase activity that was independent of water CO(2) level. Copper caused an increase in branchial metallothionein concentration that was independent of water CO(2) level. Thus, branchial metallothionein and Na(+)/K(+)-ATPase were effective biomarkers of copper exposure that were not affected by water CO(2) level.