Efficacy of Different Salt (NaCl) Concentrations in the Treatment of Ichthyophthirius multifiliis-Infected Silver Catfish,Rhamdia quelen,Fingerlings

ABSTRACT

The objective of this study was to evaluate different salt concentrations in the treatment of Ichthyophthirius multifiliis. Silver catfish, Rhamdia quelen, fingerlings infected with I. multifiliis were submitted to four sodium chloride concentrations (g/L): 0,1,2, and 4 for 23 days. In a second experiment, fingerlings were maintained with 4 g/L salt, but for 45 days. Treatments with 2 and 4 g/L salt significantly increased fingerling survival compared with 0 and 1 g/L, and survival was significantly correlated with salt concentrations. In the second experiment, fish maintained at 4 g/L salt showed a gradual reduction of “white spots” and survival was 100%. Therefore, salt offers an alternative for treatment of silver catfish fingerlings infected with I. multifiliis.     

Effects of Water pH and Hardness on Infection of Silver Catfish,Rhamdia quelen,Fingerlings by Ichthyophthirius multifiliis

The aim of this study is to determine the effect of (1) different pH levels (5, 6, 7, 8, and 9) with water hardness 20 mg/L CaCO₃ and (2) pH 5 and 7 with varying water hardnesses (20, 60, and 120 mg/L CaCO₃) on the intensity of infection by Ichthyophthirius multifiliis in fingerlings of the silver catfish, Rhamdia quelen. In Experiment 1, the lowest and highest mortality rates were observed in fingerlings exposed to pH 5 and 9, respectively; in Experiment 2, fish kept at pH 5 and 7 with hardness 20 mg/L CaCO₃ showed significantly lowest and highest cumulative mortality, respectively, than those maintained in other conditions. The lowest intensity of trophonts/fingerling was observed at pH 5 and hardness 20 mg/L CaCO₃ in both experiments. We conclude that infection by I. multifiliis is less severe in silver catfish maintained at pH 5 and hardness 20 mg/L CaCO₃. High water hardness led to an increased intensity of trophonts and decreased survival in silver catfish kept at pH 5, but decreased intensity of trophonts and improved survival when the fish were kept at pH 7.     

Toxic effects of mercury chloride on silver catfish (Rhamdia quelen) spermatozoa

Heavy metals are highly toxic elements that are present in the environment, especially in water. Mercury chloride (HgCl₂) stands out among these compounds because of its strong ability to induce damage to any tissue with which it comes into contact. The gametes of spawning aquatic animals, such as fish, are susceptible to such damage. Thus, our objective was to evaluate the toxic potential of HgCl₂ in the capacitation and activation of Rhamdia quelen sperm. Semen was collected from seven males and activated in 58 mM sodium chloride (NaCl) containing 0 (control), 4^?10, 7^?10, 7^?9, and 7^?8 M HgCl₂. The evaluated variables included motility, vigor, motility time, morphology, membrane integrity, membrane fluidity, mitochondrial functionality, production of reactive oxygen species (ROS), and DNA fragmentation. All evaluated HgCl₂ concentrations increased primary pathologies and reduced motility, vigor and motility time. Damage to membrane integrity and fluidity began occurring at a concentration of 7^?10 M HgCl₂. These results indicate that HgCl₂ has a toxic effect on different sites of fish spermatozoa and that sperm motility decreases after exposure to HgCl₂, impairing sperm capacitation and activation.     

Evaluation of Electrical Stunning of Atlantic Cod (Gadus morhua) and Turbot (Psetta maxima) in Seawater

The aim of this study was to assess electrical stunning of Atlantic cod and turbot in seawater to develop a protocol for the process of stunning and killing. An induced general epileptiform insult (unconscious) had a duration of 40 ± 27 s (n =14) in cod (2.6 ± 0.5 kg) and 34 ± 18 s (n = 19) in turbot (520 ± 65 g). Seven cod and 3 turbot displayed a physical reaction, and 11 turbot registered an electroencephalogram (EEG) response to pain stimuli administered 30 s post-stun. The heart rate was 32 ± 6 beats/min in cod and 25 ± 7 beats/min in turbot prior to stunning. Post-stunning, the electrocardiogram (ECG) revealed fibrillation and reduced activity post-stun. EEG, ECG recordings, and behavioral observations indicate that when a bipolar square wave current was applied with a frequency of 133 Hz and 43% duty cycle side to side (turbot) and at 170 Hz and 33% duty cycle (cod) head to tail, both species were stunned in seawater at current densities of 3.2 A/dm² and 2.5 A/dm², respectively. For turbot, a 5 s exposure to electricity followed by chilling in ice water for 15 min is sufficient to prevent recovery. For cod, a killing method needs to be established.     

Tilapia protein hydrolyzate improves growth performance,protein absorption and antioxidant status in Silver catfish (Rhamdia quelen)

This study evaluated the substitution of fish meal with tilapia protein hydrolyzate (TPH) in diets for Silver catfish (Rhamdia quelen) by verifying the effects on growth, body composition, blood and biochemical parameters, and biomarkers of oxidative stress. Five isoproteic and isoenergetic diets (37% crude protein, 3,800 kcal/kg) were produced with different substitution levels of TPH (0%, 5%, 10%, 15% and 20%) for fishmeal. The hydrolyzate was obtained by enzymatic hydrolysis from carcasses of Nile tilapia (Oreochromis niloticus), which were exposed to swine pepsin enzyme at 36°C for four hours. The experimental design was completely randomised with five treatments and four replications. The inclusion of 20% TPH in diets for silver catfish increased the growth of Silver catfish juveniles. At levels above 5%, there was an increase in body protein deposition and the efficiency of oxidative metabolism.     

Evaluation of head-only and head-to-tail electrical stunning of farmed eels (Anguilla anguilla, L.) for the development of a humane slaughter method

The overall objective was to evaluate the suitability of electronarcosis as a stunning method for farmed eels. In the first experiment the minimum electrical current needed to induce a general epileptiform insult by head‐only stunning was assessed. The individual eels (n = 40) with a live weight of 700–800 g were fixed in a specially designed re‐strainer. The EEG (electroencephalogram) and ECG (electrocardiogram) recordings, observation of behaviour and responses to pain stimuli were used to assess unconsciousness, insensibility and cardiac function. The applied current of 150, 200 or 250 V, 50 Hz, AC was delivered via scissor‐model stunning tongs for approximately 1 s. A general epileptiform insult was observed in 31 eels for which a successful EEG recording was obtained, using 255 ± 4 V, 545 ± 32 mA, for 1.2 ± 0.2 s. The general epileptiform insult as measured on the EEG was characterized by a tonic/clonic phase and an exhaustion phase. The behaviour showed one phase: tonic cramps alternated by clonic ones. The heart rate was 22 ± 8 beats min^?1 (n = 23) prior to stunning. After stunning the ECG revealed fibrillation. In the second experiment the behaviour of seven individual eels able to move freely in water was observed after head‐only stunning (250 V). Two phases were distinguished. Limited tonic and clonic cramps combined with backward swimming were followed by heavy clonic cramps combined with unco‐ordinated movements such as jumping out of the water. A distinct exhaustion phase was not observed in all animals. In the third experiment a head‐to‐tail electrical method was examined in 15 eels for rendering the eels unconscious and insensitive prior to slaughter. They were stunned by applying 253 V for 3 s followed by 50 V for 5 min. In the fourth experiment nine eels were head‐only stunned with 260 V for 1 s immediately followed by 50 V for 5 min applied from head to tail. Results obtained in these two experiments were similar. After stunning no brain activity and no responses to pain stimuli on the EEG were observed and the ECG showed ventricular extra systolae. It was observed that it might take 60 ± 25 min or longer for a complete recovery. It can be concluded that for effective electrical stunning of eels with a weight of 700–800 g an average current of 545 ± 32 mA (at approximate 250 V, 50 Hz AC) is needed. In this case, within a confidence level of 95% at least 91% of the eels are effectively stunned (n = 31). Therefore, it is recommended to increase the minimum current for an effective stun in practice to 600 mA. Further research is needed to determine the conditions to induce permanent unconsciousness and insensibility of the eels to protect the animals at slaughter.     

The autochthonous microbiota of the freshwater omnivores jundiá (Rhamdia quelen) and tilapia (Oreochromis niloticus) and the effect of dietary carbohydrates

Previous studies have shown that the gut microbiota of fish differs depending on host feeding habits and these populations may also be influenced by dietary ingredients. In this study, the bacterial populations of the intestinal tract of the opportunistic omnivore jundiá Rhamdia quelen and the typical omnivore tilapia Oreochromis niloticus were investigated in two experiments. In experiment I, the levels of amylolytic, cellulolytic, lipolytic, proteolytic and total culturable bacteria were compared in the proximal, mid and distal intestine using selective agar. Higher levels of amylolytic, cellulolytic, lipolytic, proteolytic and total culturable bacteria were observed in the tilapia than jundiá, in all intestinal regions. Jundiá contained higher levels of proteolytic bacteria and lipid digesting bacteria in the distal intestinal portion as compared with the mid intestine; both fish species also presented more amylolytic bacteria in the distal intestine. In experiment II, the amylolytic intestinal microbiota between the two species was compared after administration of diets containing different carbohydrate sources. Jundiá fed broken rice presented higher total culturable bacterial levels; however, dietary cassava bagasse and ground corn significantly elevated the population of amylolytic bacteria in tilapia (P > 0.05). PCR‐DGGE was also used to assess the bacterial communities in experiment II. A Cetobacterium spp. was detected in jundiá fed diets containing broken rice, and tilapia fed cassava bagasse, dextrin, broken rice and ground corn. Microbial differentiation was further demonstrated between jundiá and tilapia, because an uncultured bacterium was unique in tilapia and an uncultured spirochete was observed only in jundiá; the presence of these bacterial species was also influenced by dietary carbohydrate sources.     

Pectic hydrolysates in the diet of silver catfish (Rhamdia quelen): Growth performance,blood and liver biochemistry,histological parameters and intestinal contents

The aim of this study was to investigate the prebiotic potential of pectin hydrolysates that were extracted from agroindustrial waste (apple pomace—AP and passion fruit peel—PFP) and were added to the diet of silver catfish (Rhamdia quelen). A 49‐day biological assay was conducted, and five test diets were evaluated: one diet was a control diet and the other four diets included pectin hydrolysates (2.5 and 5 g/kg). At 49 experimental days, biometric data and biological material were collected to determine the performance, plasma and liver and histological parameters and to evaluate the intestinal contents. The results were analysed by the normality test, which was followed by an analysis of the variance; the treatment means were compared by an orthogonal contrast analysis at a 5% level of significance. The inclusion of 2.5 g/kg apple pomace hydrolysates resulted in a greater production of butyric acid, increased thickness of the muscular layer and higher goblet cell count in the intestine. The inclusion of 5 g/kg apple pomace hydrolysates led to a greater concentration of liver protein. Further studies are needed to increase the knowledge about the use of these additives in the diet of silver catfish and to establish levels that allow greater gains for the species.     

Effects of rice bran protein concentrate on the growth performance and digestive enzyme activities of jundiá (Rhamdia quelen) (Quoy and Gaimard, 1824)

The aim of this study was to assess the effect of increasing inclusion levels (100, 150, 200 and 300 g/kg) of rice bran protein concentrate (RBPC) in jundiá (Rhamdia quelen) diets using growth, body composition, somatic indices and digestive enzyme activity as parameters. Five isoproteic (370.08 ± 0.04 g/kg) and isocaloric (13.38 ± 0.04 MJ/kg) diets were formulated with four replicates per treatment. After acclimation, 500 jundiá juveniles (initial mean weight of 6.28 ± 0.12 g) were distributed into 20 round polyethylene tanks (280 L) (25 fish per tank) coupled to a thermoregulated water recirculating system. The fish were fed experimental diets three times daily (at 9:00, 13:00 and 17:00 hr) to apparent satiation. At the end of the trial (45 days), no significant differences were found in the body chemical composition, somatic indices, and trypsin and chymotrypsin enzymes of the fish fed experimental diets. A lower final weight and a lower condition factor were found in fish fed diets RBPC10 (100 g/kg of RBPC) and RBPC15 (150 g/kg of RBPC). Based on the results of this study, it is clear that the use of RBPC (at high dietary inclusion levels of 200 and 300 g kg^?1) is an effective alternative protein source to fishmeal.     

Effect of different dietary lipid levels on the reproduction of Rhamdia quelen (Quoy and Gaimard, 1824)

Lipids include some of the most important nutrients that affect the survival and growth of fishes in their early life stages. Lipid deficiency prior to spawning may significantly reduce egg production, hatchability and the number of surviving larvae. In this study, we investigated the effects of isocaloric diets containing 80, 140 and 200 g kg⁻¹ of lipids (lipid source, soy oil) for 90 days. The following data were collected at 0, 45 and 90 days: final weight; length; conditioning factor; hepatosomatic, gonadosomatic and visceral fat indices; total plasma protein; testosterone; 17ß‐estradiol; free amino acids; and ovary and muscle fatty acid profiles. Additionally, fecundity, oocyte production, egg morphometric parameters, and larval and postlarval growth were evaluated. Results were not statistically different for husbandry and biochemical parameters, but visceral fat content increased with increase in dietary lipid levels. The fatty acid profiles and composition differed among dietary treatments. The egg diameter and area were significantly low in fish fed the 200 g kg⁻¹ lipid diet, thereby hindering growth, survival and weight compared to those in postlarval fish (P < 0.05). The best reproductive rates were obtained when using diets containing 80–140 g kg⁻¹ total lipids.     

Comparison of Stunning Methods on the Physicochemical Properties of Frozen Nile Tilapia (Oreochromis niloticus) Fillets

This study compares how CO₂ narcosis and thermal shock affect the physicochemical aspects of Nile tilapia fillets held frozen (?18°C) for up to 6 months. The characteristics of tilapia fillets subjected to CO₂ narcosis were those of higher lightness and whiteness and lower redness. The hardness values of fillets of tilapia subjected to thermal shock were lower than those subjected to CO₂ narcosis. During frozen storage to both treatments, the drip loss, denaturation of myosin, and lipid oxidation increased, while fillet water holding capacity and hardness decreased. The physicochemical properties of frozen fillets of tilapia stunned by CO₂ narcosis were considered more desirable; therefore, the method can be a potential replacement to thermal shock.     

Temperature and storage period over spermatic parameters of jundiá, Rhamdia quelen (Quoy & Gaimard, 1824)

The aim of this work was to evaluate the spermatic parameters of jundia, Rhamdia quelen, semen stored for short periods under different temperatures. Fifteen males were used in a time factorial experimental design (13 × 3 × 3 × 3). The sperm was stored at: 15; 25 and 35°C and activated in water at: 15; 25 and 35°C each, respectively, within storage periods of: 0; 2; 4; 6; 8; 10; 12; 16; 20; 24; 32; 40 and 48 h after collection. The treatments were performed in triplicates and in sequential protocols every 50 h. The motility parameters were evaluated using Pearson's correlation analysis and the significant parameters (P < 0.05) were summarized using principal component analysis forming two predicted groups. Group 01 composed the following parameters: curvilinear velocity, wobble and linearity; group 02 comprised average path velocity, straight line velocity, beating cross frequency and motility rate. Effects (P < 0.05) of treatments were observed only in group 02, with interaction between time and temperature of exposure and between exposure time and water temperature. Superior results of spermatic parameters of group 02 were observed right after collection. However, the exposure and spermatic activation at 15°C assured significant results 48 h after collection when compared with 25 and 35°C.     

Effect of electrical stunning at slaughter on the carcass, flesh and eating quality of farmed sea bass (Dicentrarchus labrax)

Food quality aspects of farmed sea bass (Dicentrarchus labrax) were compared following two methods of slaughter: the normal commercial method of killing, by immersion in an ice slurry, or by first electrically stunning the fish, before immersion in an ice slurry. Quality was assessed for up to 10 days of storage on ice after slaughter. No differences were found between the slaughter methods in terms of an overall sensory evaluation of cooked fillets, or in terms of overall carcass quality: overall appearance, internal and external haemorrhage, fin damage, burst gall bladder, staining of the body cavity by leakage from the gut or damage to the spine. Using objective measurements of colour, no differences between fish from either treatment were found in terms of external colour or colour of the fillets. A chemical analysis of flesh nucleotide breakdown products as well as the freshness indicator K_i value did not differentiate the two treatments nor did the industry standard freshness scoring technique (QIM, quality index method), over 10 days of storage on ice. Flesh pH was marginally lower in electrically stunned fish at 4 h post mortem (6.42 cf 6.56) but by 24 h, pH in fish from both treatments had decreased to a similar level (6.22). Humane electrical stunning of sea bass at slaughter neither measurably improved nor decreased product quality for between 1 and 10 days of storage on ice. Electrical stunning accelerated the pattern of onset and resolution of rigor mortis. If electrical stunning were to be widely adopted, re‐education of buyers would be necessary as rigor mortis is currently used by buyers as a proxy measure of fish freshness.     

Effects of Dietary Fibre Concentrates on growth performance and digestive enzyme activities of jundiá (Rhamdia quelen)

This study was conducted to investigate the prebiotic effect of different Dietary Fibre Concentrates (DFC) (Mucilage =MG; Pectin = PN or β‐glucan + mannan = βg + M) on growth and somatic parameters, body composition and digestive enzyme activities of jundiá (Rhamdia quelen). After acclimation, fish (7.16 ± 0.06 g) were allocated into 24 tanks (30 fish per tank) and triplicate groups were fed with Control diet (0 g kg^?1 of DFC); diet supplemented with 5 g kg^?1 commercial prebiotic (CP) or diets supplemented with 5 or 10 g kg^?1 diet of MG; PN or βg + M. At the end of the trial (8 weeks), growth was significantly (P < 0.05) higher in fish fed diets supplemented with DFCs and did not differ from animals supplemented with CP. The animals that were fed Control diet presented a body protein content higher compared to those supplemented with diets containing pectin or β‐glucan + mannan (P < 0.05). However, fish fed diets added with β‐glucan + mannan yielded a higher level of protein deposited in the whole body. The activity of digestive enzymes was lower in the group supplemented with Pectin. Results indicate that supplementation with DFCs in the diet had positive effects on the performance of jundiá and are prebiotic potential candidate.     

Silver catfish Rhamdia quelen immersion anaesthesia with essential oil of Aloysia triphylla (L'Hérit) Britton or tricaine methanesulfonate: effect on stress response and antioxidant status

Responses to anaesthesia with essential oil (EO) of Aloysia triphylla (135 and 180 mg L^?1) and tricaine methanesulfonate (MS222) (150 and 300 mg L^?1) were assessed in silver catfish. Exposure to the anaesthetics elicited a stress response in the species. In the case of MS222, it was displayed as a release of cortisol into bloodstream, elevation in hematocrit and plasma ion loss. The EO presented cortisol‐blocking properties, but increased haematocrit and disturbances of hydromineral balance were observed. Liver antioxidant/oxidant status of EO and MS222‐anaesthetized silver catfish was also estimated. The synthetic anaesthetic induced lipoperoxidation, notwithstanding increased catalase contents, whereas the naturally occurring product was capable of preventing the formation of lipid peroxides, possibly due to combined actions of catalase and glutathione‐S‐transferase. Anaesthetic efficacy was also tested via induction and recovery times. Overall, the promising results obtained for the physiological parameters of the EO‐treated fish counterbalanced the slight prolonged induction time observed for 180 mg L^?1. As for 135 mg L^?1, both induction and recovery times were lengthy; despite that, the EO was able to promote oxidative protection and mitigate stress. None of the MS222 concentrations prompted such responses concomitantly.     

Field evaluation of live chilling with CO2 on stunning Atlantic salmon (Salmo salar) and the subsequent effect on quality

Atlantic salmon were slaughtered in three ways on a commercial slaughter line: (1) killed by a percussive stun after crowding; (2) killed by percussive stun after crowding, pumping and live chilling; (3) killed by exsanguination after crowding, pumping and live chilling. The live‐chilled fish were exposed to seawater (2°C) saturated with carbon dioxide (pH 5.5–5.7) for 40 min. The fish were calm after live chilling, but not unconscious, as eye rolling was observed in all individuals. Subsequent exsanguination of the unstunned fish resulted in death. Both rapid live chilling and the subsequent exsanguination appeared stressful to the fish, as a large and rapid pH drop coupled with earlier onset of rigor mortis, indicative of high muscle activity during the process were observed. The muscle core temperature during ice storage showed that live chilling only has an effect on carcass temperature during the first 6 h post mortem. After 6 h, no significant differences in temperature were detected between live‐chilled and traditionally ice‐chilled fish. We conclude that commercial use of live chilling in combination with high levels of CO₂ does not stun Atlantic salmon. Live chilling followed by exsanguination of the unstunned fish appears to be highly stressful and should be avoided.     

Responses of circulating arginine vasotocin, isotocin, and melatonin to osmotic and disturbance stress in rainbow trout (Oncorhynchus mykiss)

In teleost fish exposed to stressors AVT has been implicated in activation of the hypothalamo-pituitary-interrenal axis. Mel has been shown to counteract several behavioural and endocrine consequences of stress in mammals. These studies were undertaken to investigate the effects of either disturbance or osmotic stress, applied either separately or simultaneously, on plasma AVT, IT and melatonin in rainbow trout (Oncorhynchus mykiss). Hormones were determined in plasma by high-performance liquid chromatography preceded by solid-phase extraction. The results showed that both forms of stress caused significant increase in plasma AVT concentration, although more pronounced elevation was observed in physically disturbed fish. Conversely, neither osmotic nor disturbance stress affected plasma IT concentration. The apparent difference in response to stress by the two close related neurohypophysial nonapeptides suggests independent mechanisms controlling their synthesis and/or release and supports the idea that only AVT plays a role in physiological response to stress. Plasma Mel level was depressed in fish subjected to disturbance stress and to both stresses applied simultaneously, an effect possibly associated with the elevation of plasma AVT concentration. Results are discussed in relation to physiological interactions between hormones.