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Cardinal Tetra, Paracheirodon Axelrodi (Schultz), in the Amazon

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Cardinal Tetra, Paracheirodon Axelrodi (Schultz), in the Amazon Aquaculture Research, 2009, 40, 157^165 doi:10.1111/j.1365-2109.2008.02077.x s Benefits of using the probiotic Efinol Lduring transportation of cardinal tetra, Paracheirodon axelrodi (Schultz), in the Amazon Levy Carvalho Gomes1, Richard Philip Brinn2, Jaydione Luiz Marcon3, Lucelle Arau¤jo Dantas1, Franmir Rodrigues Brandaìo1,JanessaSampaiodeAbreu4, Paulo Evandro Mendes Lemos3, Dawn Michelle McComb5 & Bernardo Baldisserotto6 1Embrapa AmazoŒ nia Ocidental, Manaus, AM, Brazil 2Florida International University, Miami, FL, USA 3Universidade Federal do Amazonas, Manaus, AM, Brazil 4Universidade Estadual Paulista, Jaboticabal, SP,Brazil 5Florida Atlantic University,Boca Raton, FL, USA 6Universidade Federal de Santa Maria, Santa Maria, RS, Brazil Correspondence: Present address: L C Gomes, Centro UniversitaŁ rio deVilaVelha, Programa de Mestrado em Ecologia de Ecossistemas, Rua ComissaŁ rio Jose¤ Dantas de Melo 21, BoaVista,VilaVelha, ES 29102-770, Brazil. E-mail: [email protected] Abstract Keywords: probiotic, survival, Amazon, orna- mental ¢sh, ion £ux, body cortisol The objective of this experiment was to test the pro- s biotic E¢nol L during transportation of cardinal tetra, Paracheirodon axelrodi (Schultz). For the trans- Introduction portation, ¢sh were distributed in 18 plastic tanks, of s which nine contained the E¢nol L (10mg L À 1;pro- The cardinal tetra, Paracheirodon axelrodi (Schultz), is biotic treatment) and the remaining had no probiotic an endemic ¢sh species from the middle Rio Negro (control treatment). Transport lasted 24 h and three Basin widely used as an ornamental ¢sh in aqua- di¡erent boxes of each treatment were sampled at 3, riums worldwide (Walker 2004). The cardinal tetra 12 and 24 h. Up to the 12-h sampling period, no sig- is the most important commercial species of orna- ni¢cant di¡erence in the survival was observed; on mental ¢sh exported from the Brazilian Amazon, the other hand, survival was higher at the end of the contributing to about 80% of exports. Some 12^15 transport (24 h) in the probiotic treatment. No signif- million wild specimens are legally exported annually icant di¡erence was found in dissolved oxygen and and accounted by the Brazilian government (Chao temperature between treatments. Conductivity, pH 2001). However, Chao and Prada-Pedreros (1995) esti- and alkalinity increased along the transport, but mate that at least 24^32 million cardinals are taken without a di¡erence between treatments. Ammonia from the Rio Negro tributaries each year. The orna- increased in all treatments, although it was signi¢- mental industry generates 60% of the total income cantly lower in the probiotic group at 24 h. Cortisol le- of the middle Rio Negro region’s economy (Chao & vels were signi¢cantly higher in all transported ¢sh Prang 1997), with between 6000 and 8000 ¢sher- when compared with the basal values. At 24 h, cortisol men involved in this activity (Chao & Prada-Pedreros levels in control ¢sh were signi¢cantly higher than 1995; Pra ng 2001). those in the probiotic treatment.With the observed re- Transportation is an important part of all phases sults, we are able to conclude that the probiotic of the export process. Cardinals are normally caught s E¢nol L is e⁄cient during cardinal transport, lowering in small streams (igarape¤ ). After this, ¢sh are typi- the mortality and helping maintain water quality pos- cally transported in 40-L À 1 plastic boxes to Barcelos, sibly by lowering metabolic wastes. a base city for the ornamental industry in the middle r 2008 The Authors Journal Compilation r 2008 Blackwell Publishing Ltd 157 s Cardinal transportation with E¢nol L LCGomeset al. Aquaculture Research, 2009, 40, 157 ^ 165 Rio Negro region, then onto exporters’ facilities in tion in metabolic wastes and reduction in stress re- Manaus and ¢nally to importing countries, mainly sponses. Therefore, the objective of this experiment s Europe, Asia and North America (Santos & Santos was to test the e⁄ciency of E¢nol L solution during 2005). According to Waichman, Silva and Marcon the transport of the cardinal tetra between Barcelos (2001), mortality during the transportation between and Manaus. Barcelos and Manaus is a negative factor in the ex- porting process, which can be attributed to the pro- blem of bad management practices and poor water Material and methods quality leading to disease outbreaks, mostly bacteria based. This work was carried out in December 2005 during a Most recently,probiotics have appeared in the mar- scienti¢c expedition to the middle Rio Negro region, ket as a method to increase the quality and health of close to Barcelos, AM, Brazil, aboard the Miss Bebel several ¢sh species during some processes involved in vessel, with the main objective of studying the biology, management (Gatesoupe 1999; Gomez-Gil, Roque & ¢shing and management of several ornamental ¢sh Turnbull 2000). These products are primarily com- captured and exported from this region. posed of highly concentrated bacteria, vitamins and Cardinals (0.032 Æ 0.011g; mean Æ SD) were cap- nutrients (Gatesoupe 1999; Verschuere, Rombaut, tured by professional ¢shermen in a small stream Sorgeloos & Verstraete 2000) and were created to sti- called igarape¤ Puxurituba. After capture, the ¢sher- mulate the immune system and help increase the men used a standard procedure for keeping the ¢sh water quality in the environment.The use of probiotic before transport to Barcelos. Fish were kept in plastic helps increase ¢sh health during stressful processes boxes containing 20 L of water before the actual such as severe changes in environmental conditions transport. Between the capture and the experimen- (Taoka, Maeda, Jo, Jeon, Bai, Lee, Yuge & Koshio tal period, ¢sh were maintained for a maximum of 2006). Carnevali, De Vivo, Sulpizio, Gioacchini, 10 days in boxes and fed with a commercial feed Olivotto, Silvi and Cresci (2006) observed an inverse for omnivorous ¢sh (32% crude protein and correlation between gut colonization with lactic acid 4180 kcal CE kg À 1 of crude energy) twice a day. Fish bacteria strain and cortisol levels; thus, Rollo, Sulpi- were starved 2 days before the transport. zio, Nardi, Silvi, Orpianesi, Caggiano, Cresci and Car- For the transport experiment,18 plastic boxes were nevali (2006) suggest that a probiotic is able to utilized with a total capacity of 40 L but ¢lled with modulate cortisol responses in ¢sh, lowering the 10L of local stream water. Fish were randomly dis- magnitude of stress response. tributed between the boxes following the procedure Most probiotics are designed to be mixed with food, used by local ¢shermen, with a ¢nal density of but recently some probiotic products have been for- 578 Æ 26 cardinals per box or 1.84 Æ 0.19 g of ¢sh mulated that can be dissolved in water and can per litre of water. The density used was in the range help during management and transport operations. (500^1000 per box) commercially used by local ¢sh- s E¢nol L is a water-soluble commercial product sold ermen. In half of the boxes, a solution of 10mg L À 1 s as a proven anti-stress formula for aquaculture probiotic E¢nol L (Bentoli Agrinutrition, Austin, TX, hatcheries, useful for larval transport and to improve USA) was utilized, while the rest were maintained the health and survival of aquatic animal larvae.This with probiotic-free water (control group). The probio- product is formulated with Bacillus subtilis, Bacillus tic concentration is in accordance with the manufac- licheniformes, Lactobacillus acidophilus and Saccharo- turer’s instructions. A 24-h transport experiment myces cerevisiae, along with selected amino acids, proceeded with the vessel simulating conditions that vitamins, minerals, free-£ow and anti-caking agents local ¢shermen use to take their ¢sh through Barcelos (calcium carbonate and silica). This combination to Manaus. Survival, water quality, net ion £ux and characterizes the product as a probiotic enriched body cortisol were monitored from three di¡erent with some nutrients. Previous results obtained by boxes of each treatment at 3,12 and 24 h of transport. Gomes, Brinn, Marcon, Dantas, Brandaìo, Abreu, Each box was sampled only once and then discarded McComb and Baldisserotto (2008) during transpor- from the experiment. tation of another Amazonian ornamental ¢sh, the For whole-body cortisol analysis, samples of un- marbled hatchet¢sh, Carnegiella strigata (Gˇnther), stressed ¢sh were caught directly from the local s with E¢nol L demonstrated that this probiotic was streams to be compared with the transported ¢sh. e⁄cient in improving the water quality by the reduc- They were caught and anaesthetized with a lethal r 2008 The Authors 158 Journal Compilation r 2008 Blackwell Publishing Ltd, Aquaculture Research, 40, 157^ 165 s Aquaculture Research, 2009, 40, 157^165 Cardinal transportation with E¢nol L LCGomeset al. concentration of benzocaine (200 mg L À 1) and im- ment groups presented homogeneous variances and mediately frozen and stored in liquid nitrogen. survival was compared using a t-test, while water During transport, ¢sh samples were taken from each quality and net ion £uxes were compared using a box at each sampling time and frozen in liquid two-way ANOVA (time and treatment as variables) and nitrogen. the Tukey test. Di¡erences between treatments and Water analysis of temperature, dissolved oxygen body cortisol at di¡erent samplings were compared (YSI 55), pH (YSI pH-100) and conductivity with a di- with unstressed ¢sh (basal values) using one-way gital meter (Bernauer, Blumenau, Brazil) was per- ANOVA and Dunnett’stest (Po0.05) and between treat- formed directly in the box.Water samples were taken ments at the same sample time using a t-test. All sta- from the box for further analysis of alkalinity by titra- tistical tests were conducted using software STATISTICA tion (American Public Health Association, American (version 5.1).
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