Anaesthetics are important in ¢sh culture to reduce handling stress and mortality. Eugenol is a promising anaesthetic because of its low cost, e⁄cacy, safety margin for ¢sh and lack of toxicity to humans. The goal of this study was to establish a protocol using eugenol as a ¢sh anaesthetic for tambaqui Colossoma macropomum (Cuvier), and provide information for regulating authorities on establishing safety dosage protocols for its use. Juvenile and sub-adult tambaqui were ¢rst individually exposed to doses of 35, 50, 65, 85, 100 or 135 mg L À 1 eugenol for 10 min. A second experiment examined the e¡ect of the duration of exposure to eugenol on the time required for recovery and survival of tambaqui. A eugenol dose of 65 mg L À1 was adequate to induce ¢sh of both sizes into a surgical anaesthetic state, and recovery time was similar for dosages up to 100 mg L À1 . Exposure to the ideal dose (65 mg L À 1 ) for up to 30 min did not cause ¢sh mortality. Fish blood glucose values were similar for all the tested eugenol doses as well as with the benzocaine control. The results show that eugenol is an e⁄cient and safe anaesthetic for tambaqui.

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Abstract.— The present study investigated the use of benzocaine as an anesthetic for juvenile Colossoma macropomum (tambaqui). In the first experiment, fish were exposed to various doses of benzocaine for 10 min at 24 C. The second experiment examined the effects of duration of exposure to 100 mg/L of benzocaine. In the third experiment, fish were exposed to 100 mg/L at temperatures of 24, 27, and 30 C. Benzocaine concentrations of 100–150 mg/L were considered ideal for quickly inducing total immobilization and fast recovery. Fish exposed to 350 mg/L benzocaine exhibited 30% mortality. No changes in hemat‐ocrit were recorded in fish exposed to different concentrations of benzocaine. Plasma glucose increased significantly when fish were exposed to benzocaine concentrations greater than 200 mg/L. Recovery time after a 30‐min exposure to 100 mg/L benzocaine was significantly greater than after an exposure for 10 and 20 min. No mortality was observed 96 h after exposure to 100 mg/L benzocaine for 10, 20, and 30 min. Dosages in the 100–150 mg/L range were effective for periods of up to 20 min of anesthesia. There was no effect of temperature on the time required for fish to lose equilibrium. However, recovery was significantly faster for fish anesthetized at 30 C. Benzocaine is an effective anesthetic agent for tambaqui juveniles, providing rapid immobilization and rapid recovery. Benzocaine is also less expensive than other available anesthetic compounds.

The effects of graded hypoxia on the physiological and biochemical responses were examined in two closely related species of cichlids of the Amazon: Astronotus crassipinnis and Symphysodon aequifasciatus. Ten fish of each species were exposed to graded hypoxia for 8 h in seven oxygen concentrations (5.92, 3.15, 1.54, 0.79, 0.60, 0.34, and 0.06 mg O(2) L(-)(1)), with the aim to evaluate hypoxia tolerance and metabolic adjustments, where plasma glucose and lactate levels, hepatic and muscle glycogen contents, and maximum enzyme activities (PK, LDH, MDH and CS) in skeletal and cardiac muscles were measured. Another experimental set was done to quantify oxygen consumption (MO(2)) and opercular movements in two oxygen concentrations. Hypoxia tolerance differed between the two species. Astronotus crassipinnis was able to tolerate anoxia for 178 min while S. aequifasciatus was able to withstand 222 min exposure in deep hypoxia (0.75 mg O(2) L(-)(1)). Suppressed MO(2) was observed during exposure to 0.34 (A. crassipinnis) and 0.79 mg O(2) L(-)(1) (S. aequifasciatus), while opercular movements increased in both species exposed to hypoxia. Higher levels of muscle and liver glycogen and larger hypoxia-induced increases in plasma glucose and lactate were observed in A. crassipinnis, which showed a higher degree of hypoxia tolerance. Changes in enzyme levels were tissue-specific and differed between species suggesting differential abilities in down-regulating oxidative pathways and increasing anaerobic metabolism. Based on the present data, we conclude that these animals are good anaerobes and highly adapted to their environment, which is allowed by their abilities to regulate metabolic pathways and adjust their enzyme levels.

The increased demand for juvenile tambaqui Colossoma macropomum for grow‐out ponds and stocking programs in the Amazon state of Brazil has increased the transportation of this species. This study was designed to determine the optimum density of juvenile tambaqui during transportation in closed containers. Fish (51.9 ± 3.3 g and 14.9 ± 0.4 cm) were packed in sealed plastic bags and transported for 10 h at four densities: 78, 156, 234, and 312 kg/m3. After transportation, fish from each density were kept in separate 500‐L tanks for 96 h. Mortality, 96‐h cumulative mortality, water quality, and blood parameters (hematocrit, plasma cortisol, and glucose) were monitored. Fish mortality after transportation was significantly lower at densities of 78 and 156 kg/m3 than at 234 and 312 kg/m3. Cumulative mortality was significantly lower at a density of 78 kg/m3. Dissolved oxygen after 10 h of transportation remained high at a density of 78 kg/m3, but reached critically low values at all other densities. Ammonia concentration was highest at the lowest density and was lower at higher densities. Carbon dioxide concentration was lowest at the density of 78 kg/m3 but higher in the other treatments. Plasma glucose and cortisol increased significantly immediately after transportation at densities of 156, 234, and 312 kg/m3, returning to control values by 24 h. The best density for juvenile tambaqui during a 10‐h transportation haul in a closed container was 78 kg/m3. At this density there was no fish mortality, water quality was kept within acceptable values, and fish were not stressed.

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