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15 out 2021
EVENTO ONLINE
Simpósio 
Internacional 
de Nutrição e 
Saúde de Peixes
ANAIS
FACULDADE DE MEDICINA VETERINÁRIA E ZOOTECNIA
DEPARTAMENTO DE MELHORAMENTO E NUTRIÇÃO ANIMAL
AquaNutri – LABORATÓRIO DE NUTRIÇÃO E SAÚDE DE PEIXESFMVZ
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
FICHA CATALOGRÁFICA ELABORADA PELA SEÇÃO TÉCNICA DE AQUISIÇÃO E TRAT AMENTO 
DA INFORMAÇÃO – 
 DIRETORIA TÉCNIC A DE BIBLIOTECA E DOCUMENTAÇÃO - UNESP - 
FCA – 
 LAGEADO – BOTUCATU (SP) 
 
 Simpósio Internacional de Nutrição e Saúde de Peixes ( 9 . : 
S612r 2021 : Botucatu, SP) 
 Anais [do] 9 . Simpósio Internacional de Nutrição e 
 Saúde de Peixes , 15 outubro de 20 21 / Orga nização: Margarida 
 Maria Barros ... [ et al .]. Botucatu: Fa culd ade de 
 Ciências Agro nômicas , UNE SP, 2021 
 52 p. : fots. color., ils . col or., gra fs. color. , 
 tabs. 
 
 
 
Disponível em: http://w.fmvz.unesp.br/#!/lista-de-
eventos/2021/9-aqua-nutri/ 
 
1. Peixe s - Nutrição. 2. Peixe s – Saúde. 3. Peixes – 
 Aditivos. 4. Nutrição animal. I. Barros, Margarida Maria 
 I I. Pezzato, Luiz Edivaldo. III. Cyrino, José Eurico 
 Possebon. IV. Furuya, Wilson Massamitu. V . Universidade 
 Estadual Paulista “Júlio d e Mesquita Filho”, Faculdade de 
 Ciências Agronômicas . V I . Título. 
 
 
 CDD 2 3 .ed. ( 639.3 ) 
 
Ana Lucia G. Kempinas CRB/8 7310 
 
 
 
 
 
 
 
 
 
Conteúdo 
 
O evento ............................................................................................................................ 3 
Público alvo ...................................................................................................................... 3 
Objetivo ............................................................................................................................ 3 
Entidade promotora .......................................................................................................... 4 
Comissão Organizadora .................................................................................................... 5 
Palestrantes ....................................................................................................................... 7 
Programa ........................................................................................................................... 8 
Resumos do IX SINSP ................................................................................................... 10 
 
 
 
3 
 
 
 
 
 
 
 
O evento 
 
Comunicamos com grande satisfação que, apesar das limitações impostas pela 
pandemia e as dificuldades que enfrentamos na Universidade, a equipe coordenadora do 
Simpósio Internacional de Nutrição e Saúde de Peixes decidiu dar continuidade à 
programação bienal iniciada no ano de 2005. Desta forma realizaremos a 9ª edição do 
SINSP no dia 15 de outubro de 2021, de forma remota. Neste dia discutiremos os 
principais desafios encontrados na produção de peixes, no que se refere à sanidade e 
estratégias nutricionais possíveis de serem utilizadas, somada à visão da indústria de 
rações. 
 
Público alvo 
 
Informamos que o evento é de natureza científica e tem por finalidade reunir 
pesquisadores de destaque na área de nutrição de peixes e áreas correlatas do Brasil e do 
exterior. Como tem ocorrido, desde a primeira edição no ano de 2005, participarão deste 
evento alunos de graduação, pós-graduação, pesquisadores e profissionais da área de 
produção e das indústrias de rações e aditivos. 
 
Objetivo 
 
O objetivo central desta reunião é discutir os principais temas que norteiam a 
nutrição e a saúde dos peixes a partir da experiência vivida no Brasil e em outros países 
e as necessidades de nosso país. O tema principal será a utilização de estratégias 
nutricionais objetivando mitigar os efeitos do estresse e consequentes problemas 
sanitários, que podem ser apontados como principais desafios do setor produtivo e das 
indústrias de rações. 
 
 
4 
 
 
 
 
 
 
 
A consolidação da piscicultura como atividade zootécnica, a exemplo da avicultura 
e suinocultura nacional, viabilizou o desenvolvimento de rações balanceadas, que têm 
sido produzidas por várias empresas de alimentos. Entretanto, pesquisadores, empresas e 
produtores sentem a necessidade de que não só as exigências nutricionais para 
crescimento dos animais sejam atendidas, mas que visem também a saúde, a exemplo das 
principais espécies de monogástricos. Este Simpósio tem a oportunidade de apresentar os 
principais aspectos que deverão ser considerados nos próximos anos pela comunidade 
científica nacional, tendo em vista as particularidades de nosso país. 
 
Entidade promotora 
 
O evento será promovido pela Faculdade de Medicina Veterinária e Zootecnia, 
Departamento de Melhoramento e Nutrição Animal e Laboratório de Nutrição e Saúde 
de Peixes - AquaNutri, UNESP, Campus de Botucatu. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
Comissão Organizadora 
 
Dra. Margarida Maria Barros 
Professora Assistente Doutora 
Departamento de Melhoramento e Nutrição Animal, AquaNutri 
UNESP, Campus de Botucatu 
 
Dr. Luiz Edivaldo Pezzato 
Professor Emérito 
Departamento de Melhoramento e Nutrição Animal, AquaNutri 
FMVZ, UNESP, Botucatu 
 
Dr. Pedro Luiz Pucci Figueiredo de Carvalho 
Pós-doutor do Departamento de Melhoramento e Nutrição Animal, AquaNutri 
FMVZ, UNESP, Botucatu 
 
M.Sc. William dos Santos Xavier 
Doutorando do Laboratório de Nutrição e Saúde de Peixes, AquaNutri 
FMVZ, UNESP, Botucatu 
 
Dr. Wilson Massamitu Furuya 
Professor Titular da Universidade Estadual Ponta Grossa - UEPG 
Ponta Grossa, PR 
 
Dr. José Eurico Possebon Cyrino 
Professor Titular da Escola Superior de Agricultura - ESALQ USP 
Piracicaba, SP 
 
 
6 
 
 
 
 
 
 
 
Dr. Bruno Cerosi 
Professor da Escola Superior de Agricultura - ESALQ USP 
Piracicaba, SP
 
7 
 
Palestrantes 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
Programa 
 
08:30 | Término: 08:50 
ABERTURA 
Diretor da FMVZ Prof. Cezinande de Meira e membros da Comissão Organizadora 
 
09:00 | Término: 09:15 
A PESQUISA EM AQUICULTURA E A CRISE NA SAÚDE MUNDIAL 
Dr. José Eurico P. Cyrino 
Escola Superior de Agricultura Luiz de Queiroz - ESALQ, USP, Piracicaba, SP 
 
09:25 | Término: 10:05 
DESAFIOS SANITÁRIOS DA PRODUÇÃO DE TILÁPIAS NO BRASIL 
M.Sc. André Scarano 
Escama Forte, Botucatu, SP 
10:05 | Término: 10:25 
Moderadora: Dra. Fabiana Pilarski 
Centro de Aquicultura da UNESP, CAUNESP, Jaboticabal, SP 
 
10:35 | Término: 11:15 
MODULAÇÃO DO SISTEMA IMUNE DE PEIXES 
Dra. Jaqueline Dalbello Biller 
Faculdade de Ciências Agrárias e Tecnológicas, UNESP, Dracena, SP 
11:15 | Término: 11:35 
Moderador: Dr. Fernando Yamamoto 
Mississippi State University, MSU, Mississippi, USA 
 
 
 
9 
 
 
 
 
 
 
 
11:35 | Término: 14:00 
INTERVALO ALMOÇO 
 
14:00 | Término: 14:40 
ÁCIDOS ORGÂNICOS EM RAÇÕES PARA PEIXES 
Dr. José Luiz Pedreira Mouriño 
Departamento de Aquicultura, Universidade Federal de Santa Catarina, UFSC, SC 
14:40 | Término: 15:00 
Moderadora: Dra. Débora Fracalossi 
Universidade Federal de Santa Catarina, UFSC, SC 
 
15:10 | Término: 15:50 
ADITIVOS MELHORADORES DE SAÚDE EM RAÇÕES PARA PEIXES 
M.Sc. João Manoel Cordeiro Alves 
Guabi Nutrição e Saúde Animal, Campinas, SP 
15:50 | Término: 16:10 
Moderador: Dr. Giovani Sampaio Gonçalves 
Instituto de Pesca, São José do Rio Preto, SP 
 
16:10 | Término: 16:30 
ENCERRAMENTOMembros da Comissão Organizadora
 
10 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Resumos do IX SINSP 
 
 
 
 
 
 
 
 
 
 
 
 
 
11 
 
 
 
 
 
 
 
Inclusions levels of L-Glutamine in diets for giant trahira juvenile (Hoplias 
lacerdae), a Neotropical carnivorous fish species. Alfredo Rubén Palomino Ramos, 
Daniel Abreu Vasconcelos Campelo, Cristiana Leonor da Silva Carneiro, Jener 
Alexandre Sampaio Zuanon, Sérgio Luis Pinto da Matta, Wilson Massamitu Furuya, Ana 
Lúcia Salaro.....................................................................................................................18 
 
Dietary supplementation with propolis (red and green) and virgin coconut oil for 
angelfish (Pterophyllum scalare) as growth promoter. Jéssica Maria Fontes Santos, 
Fabricio Sa de Santana, Hugo Leandro dos Santos, Amanda Silva Carvalho, Shirley Avila 
Meneses, Peterson Emmanuel Guimarães Paixão, Cindy Caroline Moura Santos, Thays 
Brito Reis Santos, Higo Andrade Abe, Rodrigo Yudi Fujimoto......................................19 
 
Effect of dietary NPZnO nanoparticle and Red Aroeira etanolic extract for Nile 
Tilapia. Amanda Silva Carvalho, Hugo Leandro dos Santos, Jéssica Maria Fontes Santos, 
Shirley Avila Meneses, Fabrício Sa de Santana, Cindy Caroline Moura Santos, Thays 
Brito Reis Santos, Peterson Emmanuel Guimarães Paixão, Higo Andrade Abe, Rodrigo 
Yudi Fujimoto..................................................................................................................20 
 
Production of a multienzyme complex with residues and by-products for use as 
additives in pisciculture. Andressa Genezini dos Santos, Milene Stefani Pereira, 
Margarida Maria Barros, Luciana Francisco Fleuri.........................................................21 
 
Digestible lysine requirement in diets for tambaquis (Colossoma macropomum) in 
the final production phase (Preliminary results). Rodrigues, Andressa Tellechea; 
Oliveira, Magdiel Santos; Campos, Denis William Johansem; Silvério, Caio Alexandre; 
Costa, Jesaías Ismael; Lima, Jéssica Pacheco; Aguilar, Gabriel da Silva; Carneiro, Dalton 
José..................................................................................................................................22 
 
 
12 
 
 
 
 
 
 
 
Metabolic responses of tambaquis (Colossoma macropomum) fed with different 
levels of lysine (Preliminary results). Rodrigues, Andressa Tellechea; Campos, Denis 
William Johansem; Oliveira, Magdiel Santos; Quiroz, Victor Alexander Cueva; Silvério, 
Caio Alexandre; Costa, Jesaías Ismael; Oshiro, Amanda Miyuki; Hiromoto, Marcelo 
Joho; Carneiro, Dalton José..............................................................................................23 
 
Zootechnical performance of red tilapia fingerlings in aquaponic system in different 
doses of probiotic + prebiotic. Barbara A. Kasuga, Cintia P. Araújo, Gabriela S. 
Carvalho, Janaína S. I. Valandro, Vinícius M. Zabotto, Eduardo A. Sanches.................24 
 
TNF-α as an indicator of immune system activation after β-glucan administration 
in Nile tilapia diets. Basia Schlichting Moromizato, Ingrid Camargo dos Reis, Laura 
Cristina de Paula, Jaqueline Dalbello Biller.....................................................................25 
 
Amino acids are more efficient than IGF1 in stimulating pathways of development 
and growth in pacu (Piaractus mesopotamicus) muscle cells. Bruna Tereza Thomazini 
Zanella, Erika Stefani Perez, Edson Assunção Mareco, Bruno Oliveira Silva Duran, Maeli 
Dal-Pai-Silva...................................................................................................................26 
 
Oregano essential oil supplementation for the catfish Lophiosilurus alexandri fed with 
high plant protein diets. Cristiana Leonor da Silva Carneiro, André Luis Souza 
Modesto, Rafael Costa Teixeira Rusth, Roger Iván Valderrama Londoño, Daniel Abreu 
Vasconcelos Campelo, Antônio Policarpo Souza Carneiro, Jener Alexandre Sampaio 
Zuanon, Ana Lúcia Salaro................................................................................................27 
 
 
 
 
 
13 
 
 
 
 
 
 
 
Antimicrobial activity of essential oils against Klebsiella pneumoniae isolated from 
diseased Nile tilapia farmed in Brazil. Daiane Vaneci-Silva, Inácio Mateus Assane, 
Fabiana Pilarski…………………………………………………………………………28 
 
Growth and survival of Nile tilapia fed diets containing nucleotides and challenged 
against Streptococcus agalactiae. Evandro Bilha Moro, Inácio Mateus Assane, Suzana 
Kotzent, Karen Dayana Prada Mejia, Bruno Luis Miani Verri, André do Vale Oliveira, 
Fabiana Pilarski................................................................................................................29 
 
Immunostimulation: The investiment in promoting fish health. Giselle Eler………30 
 
Resistance of Nile Tilapia supplemented with zinc oxide nanoparticle and red 
aroeira extract against Streptococcus agalactiae. Hugo Leandro dos Santos, Amanda 
Silva Carvalho, Jéssica Maria Fontes Santos, Shirley Ávila Meneses, Fabrício Sa de 
Santana, Cindy Caroline Moura Santos, Thays Brito Reis Santos, Peterson Emmanuel 
Guimarães Paixão, Higo Andrade Abe, Rodrigo Yudi Fujimoto......................................31 
 
Propolis (red and green) and virgin coconut oil as immunostimulant for angelfish 
(Pterophyllum scalare). Fabricio Sa de Santana, Jéssica Maria Fontes Santos, Amanda 
Silva Carvalho, Hugo Leandro dos Santos, Shirley Avila Meneses, Cindy Caroline Moura 
Santos, Thays Brito Reis Santos, Peterson Emmanuel Guimarães Paixão, Higo Andrade 
Abe, Rodrigo Yudi Fujimoto............................................................................................32 
 
β-glucan stimulates the expression of interleukin 10 (IL-10) and immunoglobulin M 
(IgM) to prevent pathogens entrance in vaccinated Nile tilapia. Ingrid Camargo dos 
Reis, Camino Fierro-Castro, Raphael Farias Mathias, Jaqueline Dalbello Biller.............33 
 
 
 
14 
 
 
 
 
 
 
 
β-glucan potentiates the expression of HsP70 gene in vaccinated Nile tilapia. Ingrid 
Camargo dos Reis, Camino Fierro-Castro, Thais Cristina de Oliveira, Jaqueline Dalbello 
Biller................................................................................................................................34 
 
β-glucan administration for 60 days stimulates the IL-1β and IL-8 gene expression 
for up to 90 days in Nile tilapia. Jaqueline Dalbello Biller, Ingrid Camargo dos Reis, 
Laura Cristina de Paula, Basia Schlichting Moromizato..................................................35 
 
Microbiome of Nile tilapia juvenile fed diets with xylanase and β- glucanase. João 
Antônio Galiotto Miranda, Johnny Martins de Brito, Paola Aparecida Paulovski 
Panaczevicz, Allan Vinicius Urbich, Angelo Machado Soltes Filho, Ana Flávia Moreira, 
Valéria Rosseto Barriviera Furuya, Wilson Massamitu Furuya.......................................36 
 
β-glucan effects on blood leukocyte respiratory burst activity. Laura Cristina de Paula, 
Ingrid Camargo dos Reis, Nathália Gabrielle Aranibar Passos, Jaqueline Dalbello 
Biller................................................................................................................................37 
 
Fermented soybean meal in the feed reduces pathogenic bacteria in South American 
Catfish (Rhamdia quelen) intestines. Luiz Augusto Cipriani, Nandara Soares de 
Oliveira, Natalia Ha, Larissa da Cunha, Thiago El Hadi Perez Fabregat.........................38 
 
Growth performance of South American Catfish (Rhamdia quelen) fed with 
fermented soybean meal. Luiz Augusto Cipriani, Nandara Soares de Oliveira, Natalia 
Ha, Larissa da Cunha, Thiago El Hadi Perez Fabregat.....................................................39 
 
 
 
 
 
15 
 
 
 
 
 
 
 
Apparent digestibility of the proteinfraction in diets containing protein levels and 
proportions of animal source protein for juvenile pacu. Oliveira, Magdiel Santos; 
Campos, Denis William Johansem; Rodrigues, Andressa Tellechea; Lima, Jessica 
Pacheco; Costa, Jesaías Ismael; Silvério, Caio Alexandre; Carneiro, Dalton José...........40 
 
Surgical procedure in spontaneous neoplasm of Carassius auratus. Maria Cecília de 
Lima Rorig, Monica Regina de Matos, Solimar Dutra da Silveira, Sara Ugulino Cardoso, 
Leonardo Aluisio Baumgartner, Jânderson Rocha Garcez, Anna Cristina Strieder 
Dalmaso, Amanda Moreira Malacarne, Thaís Voelkl Chagas, Micáila Bolzon Gonzalez 
e Robie Allan Bombardelli...............................................................................................41 
 
Effects of orange peel fragment, vitamins C and E, and zinc supplementation on 
antioxidant enzyme activity of Nile tilapia (Oreochromis niloticus) subjected to 
transport-induced stress. Igor Simões Tiagua Vicente, Matheus Gardim Guimarães, 
Paulo Incane Ito, William dos Santos Xavier, Edgar Junio Rodrigues Damasceno, Pedro 
Luiz Pucci Figueiredo de Carvalho, Lucas Franco Miranda Ribeiro, Gabriel Alighieri de 
Todos os Santos Vieira Loureiro, Luiz Edivaldo Pezzato, Margarida Maria Barros........42 
 
Effect of prolonged β-glucan administration on the hematology of tilapia, 
Oreochromis niloticus. Milene Gomes Vieira da Silva, Simone Andréia Assis de Beneti, 
Flaviane Wanessa Lopes Fernandes, Jaqueline Dalbello Biller........................................43 
 
Insect meals for pacu (Piaractus mesopotamicus) fingerlings: somatic indices and 
weight gain. Santos, Monique Virães B.; Silva Neto, Manuel R.; Costa, Jayne R.; Lima, 
Cristian J. B.; Gonçalves, Giovani S.; Fernandes, João B. K............................................44 
 
Evaluation of commercial elasmobranch supplements. Murilo José Marques Maia, 
Juliana Almeida Formágio, Julia Medeiros Mercado.......................................................45 
 
16 
 
 
 
 
 
 
 
Evaluation of glucose levels in different lineages of Nile Tilapia feed with different 
fonts of oils. Pedro Massahiro de Matos Murata, Diana Carla Fernandes Oliveira, 
Jefferson Gomes Clementino, Rilke Tadeu de Freitas......................................................46 
 
Inclusion of natural additives in diet of tambaqui fingerlings (Colossoma 
macropomum, Cuvier, 1818). Félix Aparecido Ramos, Wesclen Vilar Nogueira, Nilson 
da Silva Mendes, Weder Carlos de Souza, Analice Timoteo de Araujo, Bruna Rafaela 
Caetano Nunes Pazdiora, Filipe dos Santos Cipriano, Ricardo Henrique Bastos de 
Souza...............................................................................................................................47 
 
Virgin coconut oil dietary supplementation and feeding deprivation for Nile Tilapia 
juveniles. Shirley Ávila Menezes, Elder Davy Oliveira do Nascimento, Amanda Silva 
Carvalho, Jéssica Maria Fontes Santos, Hugo Leandro dos Santos, Fabrício Sa de 
Santana, Cindy Caroline Moura Santos, Thays Brito Reis Santos, Peterson Emmanuel 
Guimarães Paixão, Higo Andrade Abe, Rodrigo Yudi Fujimoto......................................48 
 
Weaning from Artemia nauplii to dry diet in lambari Astyanax lacustris intensive 
larviculture. Shisley Cristina da Silva Manso, Rosangela Kiyoko Jomori……………..49 
 
Xylanase and β-glucanase on growth performance, activity of digestive enzymes, 
blood parameters and digestibility in juvenile Nile tilapia fed diets with soybean 
meal and distillers dried grains with solubles from corn. Thais Pereira da Cruz, Élison 
Silva de Macêdo, Analene Alves Nascimento, Allan Vinnicius Urbich, Paola Aparecida 
Paulovski Panaczevicz, João Antônio Galiotto Miranda, Valéria Rossetto Barriviera 
Furuya, Wilson Massamitu Furuya..................................................................................50 
 
Degradation of aflatoxin B1 in fish feed by enzymatic action. Wesclen Vilar Nogueira, 
Marcelo Borges Tesser, Jaqueline Garda Buffon.............................................................51 
 
17 
 
 
 
 
 
 
 
Orange peel fragment as substrate for solid-state fermentation: a potential source 
of lipase for aquafeeds. William dos Santos Xavier, Milene Stefani Pereira, Ana Clara 
Candido Azarias, Gabriel Antunes Ferrarezi, Edgar Junio Rodrigues Damasceno, 
Matheus Gardim Guimarães, Paulo Incane Ito, Pedro Luiz Pucci Figueiredo de Carvalho, 
Luciana Francisco Fleuri, Luiz Edivaldo Pezzato, Margarida Maria Barros....................52 
 
 
 
 
 
 
 
 
 
 
 
Inclusions levels of L-Glutamine in diets for giant trahira juvenile (Hoplias lacerdae), a 
Neotropical carnivorous fish species 
 
Alfredo Rubén Palomino Ramos*, Daniel Abreu Vasconcelos Campelo1, Cristiana Leonor da 
Silva Carneiro2, Jener Alexandre Sampaio Zuanon2, Sérgio Luis Pinto da Matta3, Wilson 
Massamitu Furuya4, Ana Lúcia Salaro2 
 
*Professor e Coordenador do curso Engenharía em Aqüicultura da Universidad Científica del Sur, Rodovía 
Panamericana Sur, Lima – Peru, apalominor@cientifica.edu.pe; 1Instituto de Estudos Costeiros (IECOS), 
Universidade Federal do Pará (UFPA), Alameda Leandro Ribeiro s/n, 68600-000 Bragança, Pará, Brasil; 
2Departamento de Biologia Animal (DBA), Universidade Federal de Viçosa (UFV), Avenida Peter Henry Rolfs s/n, 
36570-900 Viçosa, Minas Gerais, Brazil; 3Departamento de Biologia Geral (DBG), Universidade Federal de Viçosa 
(UFV), Avenida Peter Henry Rolfs s/n, 36570-900 Viçosa, Minas Gerais, Brasil;4Universidade Estadual de Ponta 
Grossa, Departamento de Zootecnia, Avenida Carlos Cavalcanti, n◦ 4748, 84030-900 Ponta Grossa, Paraná, Brasil. 
 
Supplementation of crystalline amino acids in diets can be an important tool in formulating diets 
that meet the amino acid requirements of fish. The importance of determining essential amino acid 
requirements is already widely discussed and evaluated for several fish species. However, non-
essential amino acids can play an important role in maintaining normal physiological functions 
and nutritional status in fish. Glutamine, a non-essential amino acid, has been considered a 
functional amino acid because of its beneficial effects on fish growth and intestinal health. 
However, to our knowledge, no researches have been conducted to evaluate L-glutamine 
supplementation on growth performance and health of giant trahira (Hoplias lacerdae), a 
neotropical carnivorous fish. The experimental design was completely randomized with six 
treatments (0.0; 2.0; 4.0; 6.0; 8.0 or 10.0 g kg-1 of L-glutamine) and five replications. Fish (1.69 ± 
0.10 and 4.67 ± 0.10 cm) were distributed in 30 circular tanks (20 L) arranged in a water 
recirculation system. The fish were fed for 12 weeks until apparent satiety at 08:00, 12:00 and 
18:00 h. Polynomial regression showed a quadratic effect (p < 0.05) of L-glutamine levels on body 
weight h gain (y = 27.349 + 3.747x – 0.388x2; R2 = 0.780; Ymax = 4.83 g kg
-1) and fish uniformity 
(y = 3.364 + 0.583x – 0.059x2; R2 = 0.670; Ymax = 4.93 g kg
-1). The whole-body composition was 
unaffected by dietary treatments (P > 0.05). According to second-order polynomial analysis, based 
on the villi height, the optimum dietary L-glutamine levels was estimated at 5.42 g kg-1 dry diet (y 
= 434.941 + 38.740x – 3.574x2; R2 = 0.690). These results would allow us to state that diets 
containing glutamine levels above the fish physiological requirements would negatively affect the 
integrity of the intestinal structure and affect digestion and absorption functions, resulting in a 
decreased growth responses of fish. Overall, dietary supplementation with L-glutamine can 
improve growth performance and intestinal histomorphometry of juvenile giant trahira, and at the 
same time, the optimal level in the diet is between 4.83 and 5.42 g kg-1. 
Key-words: absorption, amino acid, growth, histology, villi 
Finance Institutions:CAPS, CNPQ and FAPEMIG 
 
18
 
 
 
Dietary supplementation with propolis (red and green) and virgin coconut oil for 
angelfish (Pterophyllum scalare) as growth promoter 
 
Jéssica Maria Fontes Santos1, Fabricio Sa de Santana1, Hugo Leandro dos Santos1, 
Amanda Silva Carvalho*, Shirley Avila Meneses1, Peterson Emmanuel Guimarães 
Paixão2, Cindy Caroline Moura Santos2, Thays Brito Reis Santos2, Higo Andrade 
Abe3, Rodrigo Yudi Fujimoto4 
 
*Graduate at Zootecnia, Federal University of Sergipe; Av. Marechal Rondom; 49100- 
000- São Cristóvão- SE; amandasc_zoo@outlook.com; 1 Federal University of Sergipe, SE; 
2Tiradentes University, SE; 3Federal University of Pará, PA; 4Brazilian Agricultural Research 
Corporation, SE 
 
In Brazil, angelfish (Pterophyllum scalare) stand out in the national ornamental fish 
market due to their colors and morphological aspect, as well as the rearing in captivity 
is easily performed. However, the production of this species have intensively increased, 
and fish farmers and researchers have been searching for feed additives to promote 
better growth performance. Particularly, propolis and virgin coconut oil has been cited 
as feed additives which improve the growth, but actually, scientific reports for angelfish 
still are missing. For this reason, this study evaluated the growth performance of 
angelfish submitted to diets containing propolis (red and green) and virgin coconut oil. 
To prepare the diets, 1g of each propolis was soaked into ethanol for 1 hour by static 
maceration at room temperature. Afterwards, its content was filtered and then sprinkled 
in the diet. The experiment occurred in completely randomized design in factorial 
arrangement (3x3) containing three levels of virgin coconut oil (0, 50 and 100% 
replacing soybean oil as lipid source), three propolis supplementation (none, green and 
red at proportion 10g/kg of ration) in triplicates. The animals (0,10±0,02g e 
1,02±0,05cm) received the trial diets throughout 90 days at feeding frequency of three 
meals a day and feeding rate of 5% of biomass. Monthly were carried out biometric 
procedures to determine weight (g), length (cm), specific growth rate (%), biomass gain 
(g), apparent feed conversion and survival. All data were submitted to analysis of 
variance (ANOVA-two way) followed by Tukey test (5%). As result, a statistic 
interaction occurred between coconut oil and propolis (red and green) for survival, and, 
higher values of survival rate (70 to 80%) were observed regardless the level of propolis 
or coconut oil. Nonetheless, separately, the virgin coconut oil at concentration 100% 
caused the lowest survival (46%). For growth performance, red and green propolis and 
100%coconut oil promoted better length and biomass gain. Diet with red propolis plus 
coconut oil at 50 or 100% promoted weight gain and apparent feed conversion. As 
conclusion, supplementation of propolis (red or green) plus virgin coconut oil in the diet 
for angelfish improves the growth performance. 
 
19
 
 
 
Effect of dietary NPZnO nanoparticle and Red Aroeira etanolic extract for Nile 
Tilapia 
 
Amanda Silva Carvalho*, Hugo Leandro dos Santos1, Jéssica Maria Fontes Santos1, 
Shirley Avila Meneses1, Fabrício Sa de Santana1, Cindy Caroline Moura Santos2, Thays 
Brito Reis Santos2, Peterson Emmanuel Guimarães Paixão2, Higo Andrade Abe3, 
Rodrigo Yudi Fujimoto4 
 
*Graduate at Zootecnia, Federal University of Sergipe; Av. Marechal Rondom; 49100-000- São Cristóvão- 
SE; amandasc_zoo@outlook.com; 1Federal University of Sergipe, SE; 2Tiradentes University, SE; 3Federal 
University of Pará, PA; 4Brazilian Agricultural Research Corporation, SE 
 
The increase in demand for high quality protein caused the intensification of tilapia rearing 
around the word. For this reason, the use of supplements and nutritional additives has been 
studied to provide better fish growth and sustainable development. Thus, different 
alternatives such as nano and phytotherapy are commonly applied due to their chemical 
characteristics, especially the zinc oxide nanoparticle (NPZnO) and red Aroeira extract. 
Therefore, this study aimed to evaluate the effect of dietary zinc oxide nanoparticle and 
alcohol extract of red Aroeira as growth promoter for Nile Tilapia. The NPZnO 
nanoparticles were synthetized by chemical method using zinc acetate dehydrate in 
magnetic shaking throughout 2 hours at room temperature. The alcohol extract of red 
Aroeira (AEA) was produced by static maceration (grinded leaf into ethanol 100%) also 
in magnetic shaking throughout 1 hour at room temperature. The basal diets received 
15mg/kg of NPZnO and 100mL/kg of AEA, following a completely randomized design 
with three treatments (none additives, NPZnO and AEA) and four replicates. A total of 
120 fish were distributed into 12 aquaria (70L) receiving four meals a day, throughout 60 
days with adjusted feeding rate per month (30 days: 10% of biomass; 60 days: 8% of 
biomass). At the end of experimental period, biometric procedures were carried out the 
estimate growth performance determining weight, length, biomass, apparent feed 
conversion, ration consumption, relative condition factor, specific growth rate and 
survival. All of data were submitted to analysis of variance with post hoc Tukey test (5%). 
As result, the diets containing NPZnO and AEA promoted higher biomass gain (135.6g 
and 115.1g respectively), and lowest values for apparent feed conversion (1.1±0.1 and 
1.1±0.1) which was influenced by the ration consumption among the treatments. The other 
parameters did not differ statistically at the end of experimental period. As conclusion, 
zinc oxide nanoparticle and the red aroeira ethanol extract promotes better growth 
performance for Nile Tilapia. 
 
20
 
 
Production of a multienzyme complex with residues and by-products for use as 
additives in pisciculture 
 
Andressa Genezini dos Santos*, Milene Stefani Pereira1, Margarida Maria Barros2, 
Luciana Francisco Fleuri1 
 
*Mestre em Zootecnia; Universidade Estadual Paulista; Rua Prof. Dr. Antônio Celso Wagner Zanin 250; 
18618-689 - Botucatu - SP; andressa.genezini@unesp.br; 1 Instituto de Biociências de Botucatu, 
Universidade Estadual Paulista, Botucatu, SP; 2 Faculdade de Medicina Veterinária e Zootecnia, 
Universidade Estadual Paulista, Botucatu, SP. 
 
Agro-industry residues and by-products have an immense availability after raw material 
processing and can be used as substrates for solid-state fermentation (SSF) for the 
purpose of producing enzymes to be used as additives in pisciculture. The orange peel 
fragment (frit) of the Pêra variety and the dry distillery grains (DDGS) of corn were used 
as substrates for SSF with three fungi Aspergillus flavipes (ATCC 16841), Aspergillus 
niger (CBMAI 2084) and Aspergillus oryzae (ATCC 1003), for the production of 
enzymes (amylase, phytase, lipase and protease). The multienzyme complex (MeC) 
DDGS fermented with A. flavipes and A. oryzae did not show fungal growth. MeC DDGS 
+ A. niger and frit + A. niger obtained statistically similar results (p ≥0.05) for the 
enzymatic activities of lipase using palmitate (35.16 and 29.72 U g-1 min-1) and of phytase 
(544.52 and 470.44 µmol g-1 min-1), respectively. MeC DDGS + A. niger showed 
superior results for the enzymatic activity of lipase by titrimetric analysis (19.25 U g-1 
min-1), as well as similar amylase activity to the value found in MeC frit + A. oryzae 
(8.62 and 6.19 µmol g-1 min-1, respectively). MeC frit + A. niger showed superior results 
for protease activity (22.67 U g-1 min-1) in relation to the other treatments. So, the 
multienzymes complexs produced using frit and DDGS through SSF with the fungus A. 
niger can be used as additives for pisciculture. 
 
Acknowledgment: CNPq. 
21
 
 
 
 
 
 
 
Digestible lysine requirement in diets for tambaquis (Colossoma macropomum) inthe final 
production phase (Preliminary results) 
 
Rodrigues, Andressa Tellechea*1; Oliveira, Magdiel Santos2; Campos, Denis William Johansem2; 
Silvério, Caio Alexandre2; Costa, Jesaías Ismael1; Lima, Jéssica Pacheco1; Aguilar, Gabriel da 
Silva1; Carneiro, Dalton José 1 2 
*1
Centro de Aquicultura; Universidade Estadual Paulista “Júlio de Mesquita Filho” (UNESP); Via de Acesso Prof. 
Paulo Donato Castellane s/n; 14884-900- Jaboticabal, SP, Brasil. E-mail: andressa.tellechea@unesp.br 
2
Departamento de Zootecnia; Faculdade de Ciências Agrárias e Veterinárias; Universidade Estadual Paulista “Júlio 
de Mesquita Filho” (UNESP); 14884-900- Jaboticabal, SP, Brasil. 
 
Meeting the nutritional requirement is necessary for adequate growth, physiological functions, 
better nitrogen utilization and deposition. The knowledge of the dietary lysine requirement in the 
different growth stages of tambaquis is extremely important from a nutritional point of view, 
especially in the intermediate stage of growth, which lacks information. Thus, the aim of the study 
was to estimate the digestible lysine requirement in diets for tambaquis (C. macropomum) in the 
final production phase (from 125 to 500g) for maximum growth. A total of 252 tambaquis (14 
fish/experimental unit) with an average initial weight of 126.19±0.76g were distributed in 18 
experimental units in a water recirculation system, following a completely randomized design with 
six treatments and three replications. Fish were fed until apparent satiety for 80 days with 
isoprotein (23.5% DP) and isocaloric (3350 kcal kg DE) diets, containing six increasing levels of 
digestible lysine: 0.82; 1.12; 1.32; 1.72; 2.02 and 2.32%. The physical chemical parameters of the 
water were measured and kept within the comfort range of the species. The determination of the 
requirement was performed based on the variable of weight gain, as a function of the minimum 
necessary level of lysine through the adjustment of data by the quadratic model associated with 
the plateau of the Linear Response Plateau (LRP), using RStudio software, version 1.4.1717. The 
digestible lysine requirement for better performance based on the variable of weight gain was 12.8 
g kg-1 for tambaquis in the final production phase (125 to 500 grams). 
 
Acknowledgments 
We thank the National Council for Scientific and Technological Development (CNPq) (Fellowship 
Process No. 140781/2019-6), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – 
Brasil (CAPES) (Finance Code No. 001) and the employee Jesaias Costa (São Paulo State 
Research Support Foundation-FAPESP, fellowship Process No. 2018/23605-9) for supporting this 
work. We are also grateful to CJ do Brasil LTDA, Guabi Nutrição e Saúde Animal S.A., Evonik 
Industries, Alltech do Brasil and Group CASSAB by the donation of inputs. 
22
 
 
 
 
 
 
 
Metabolic responses of tambaquis (Colossoma macropomum) fed with different levels of 
lysine (Preliminary results) 
 
Rodrigues, Andressa Tellechea*1; Campos, Denis William Johansem2; Oliveira, Magdiel Santos2 ; 
Quiroz, Victor Alexander Cueva1; Silvério, Caio Alexandre2; Costa, Jesaías Ismael1; Oshiro, 
Amanda Miyuki1; Hiromoto, Marcelo Joho1; Carneiro, Dalton José 1 2 
*1Centro de Aquicultura; Universidade Estadual Paulista “Júlio de Mesquita Filho” (UNESP); Via de Acesso Prof. 
Paulo Donato Castellane s/n; 14884-900- Jaboticabal, SP, Brasil. E-mail: andressa.tellechea@unesp.br 
2Departamento de Zootecnia; Faculdade de Ciências Agrárias e Veterinárias; Universidade Estadual Paulista “Júlio 
de Mesquita Filho” (UNESP); 14884-900- Jaboticabal, SP, Brasil. 
 
Lysine is the precursor of L-carnitine synthesis, it has the function of transporting long-chain fatty 
acids to the mitochondria for energy production through the β oxidation of lipids. The adequate 
supply of lysine to meet the needs of a species is particularly important to avoid physiological and 
growth alterations. Therefore, studies on the influence of this amino acid, linked to the specific 
characteristics of the metabolism of tambaquis, become equally important. Thus, the aim of the 
study was to evaluate the effect of lysine levels on blood metabolic variables on the growth of 
tambaquis (from 125 to 500g) fed diets containing different levels of digestible lysine. A total of 
252 tambaquis (14 fish/experimental unit), with an initial average weight of 126.19±0.76g were 
distributed in 18 experimental units (volume of 1000L), following a completely randomized design 
with six treatments and three replications. Fish were fed until apparent satiety for 80 days, with 
isoprotein (23.5% DP) and isocaloric (3350 kcal kg DE) diets, containing six increasing levels of 
digestible lysine: 0.82; 1.12; 1.32; 1.72; 2.02 and 2.32%. At the end of the experimental period, 
fish blood was collected by tail vein puncture for the determination of plasma glucose and 
triglycerides, and serum concentrations of total protein, albumin and total cholesterol. Blood 
variables were analyzed by spectrophotometry using commercial kits. Analysis of variance (One-
way ANOVA) and comparison of means by Tukey's test (P<0.05) revealed that the highest mean 
values of final weight (542.29±6.19 g), total protein (3.69±0.05 g/dL) and total cholesterol 
(193.81±26.86 mg/dL) were obtained with the diet containing 1.42% Lys. The diet without amino 
acid supplementation (0.82% Lys) provided the lowest values of total protein (3.40±0.21 g/dL) 
and glucose (113.55±6.42 mmol/L-1). Fish fed the diet containing 1.12% Lys had the lowest values 
of total cholesterol (170.91±0.67 mg/dL) and triglycerides (417.13±30.39 mg/dL). The mean 
albumin levels did not differ between treatments (P<0.05), with an overall mean of 1.08±0.02 
g/dL. Fish fed with lysine contents higher than 1.42% exhibited higher levels of total protein 
(greater than 3.50±0.19 g/dL), glucose (greater than 115.07±1.69 mmol/L-1) and triglycer ides 
(greater than 477.01±30.39 mg/dL), when compared to fish fed diets containing 0.82 and 1.12% 
digestible lysine. Lysine levels between 1.12 and 2.32% generated high levels of serum proteins, 
which act as growth performance predictors and exacerbated increases in blood cholesterol. 
Acknowledgments: CNPq (Fellowship Process No. 140781/2019-6) and CAPES (Code No. 001). 
23
 
 
Zootechnical performance of red tilapia fingerlings in aquaponic system in 
different doses of probiotic + prebiotic. 
 
Barbara A. Kasuga1*, Cintia P. Araújo2, Gabriela S. Carvalho2, Janaína S. I. 
Valandro1, Vinícius M. Zabotto2, Eduardo A. Sanches1, 2 
 
Unesp Aquaculture Center, Caunesp, Jaboticabal Campus, Access Road “Prof. Paulo Donato 
Castellani”, Km 5, 14888-000, Jaboticabal-São Paulo, Brazil. In: barbara.ayumi@unesp.br. 
 
Studies with more sustainable and alternative systems that reduce the use of 
chemotherapy in fish farms have increased in recent years due to the intensification of 
the aquaculture sector. The probiotic associated with prebiotic is a compound used to 
improve water quality and zootechnical performance of fishes. In this way, the objective 
was to evaluate the zootechnical performance of red tilapia fingerlings in aquaponics 
system under inclusion of increasing probiotic levels. The experiment was assembled in 
a completely randomized design with four treatments (0, 0.05, 0.20, 0.35 g/m3 of 
probiotic DBAqua® in water) and four replications. Were used 480 red tilapia 
fingerlings (37.35±14.03 g of average weight) and distributed in 16 polyethylene boxes 
of 310 L (300 L useful) in a closed recirculation system containing 20 lettuce seedling 
each. During a 40 days period, the fish were fed with 2% of the biomass, and offered 
four times a day with the commercial extruded feed containing 32% CP, during the 
autumn and winter period (18±2.71º C of the average water temperature). Water quality 
parameters were performed once a week. At the end of the experiment,the fishes were 
collected, anesthetized, weighed, and measured to evaluate zootechnical indices. The 
parameters obtained were submitted to analysis of variance of one factor (one-way 
ANOVA) at 5% significance and in case of significant effect were submitted to 
comparison analysis of Tukey's media. Weight (47.37±2.49 g), average weight gain 
(10.72±2.16 g), biomass (1294.63±127.50 g), biomass gain (1101.71±12.22 g), daily 
weight gain (0.27±0.0 5 g), apparent feed conversion (2.17±0.56%), specific growth 
rate (0.64±0.11 %), daily live feed intake (0.33±0.03 %-1), feed efficiency 
(48.48±9.82%), survival (91.67±9.19 %), were not significant in the doses of probiotics 
(p>0.05) in the experimental units. There were no significant differences (p>0.05) 
between the analyses of total ammonia (mg L-1), nitrite (mg L-1), dissolved oxygen (mg 
L-1), pH, conductivity (uS) and temperature (ºC) throughout the experiment. In 
conclusion, the use of probiotics + prebiotic can be used up to 0.35 g/m3, once the 
probiotic does not interfere in the zootechnical performance of juveniles when worked 
in aquaponic system. 
 
I thank CAPES for the financial and institutional support 
 
 
24
 
 
TNF-α as an indicator of immune system activation after β-glucan administration 
in Nile tilapia diets. 
 
Basia Schlichting Moromizato1*, Ingrid Camargo dos Reis1, Laura Cristina de Paula1, 
Jaqueline Dalbello Biller1 
*¹ Faculdade de Ciências Agrárias e Tecnológicas, FCAT-UNESP, Campus de Dracena, Brasil. 
basia.moromizato@unesp.br 
 
The immunology of aquatic organisms is a topic of great importance worldwide, since 
the intensification of aquaculture has led to the emergence of diseases and, 
consequently, the growing interest in the immune system. Immunostimulants, among 
them, β-glucan is one of the most used and studied compounds in fish. TNF-α is a 
cytokine from a specific group of proteins produced by different cell types, with low 
molecular weight, and which are capable of modulating the response of different cells. 
They are produced in order to activate, mediate or regulate the total immune response 
and depend on specific receptor-substrate binding. That is, they only activate cells that 
have the specific receptor. Cytokines are closely related to the inflammatory process. 
Thus, the aim of the present study was to evaluate the effects of β-glucan on the 
expression of the TNF-α gene. The experiment was conducted in a completely 
randomized design with three trials and was designed to assess the effect of β-glucan 
over time. Thus, tilapia were fed a control diet and an experimental diet with 10g kg-1 
of β-glucan for 30, 60 and 90 days. In the first trial (30 days of β-glucan), fish were fed 
either the control diet or the β-glucan diet for 30 days and then sampled. In the second 
trial (60 days of β-glucan), fish were fed control diet or β-glucan for 60 days and β-
glucan diet for 30 days plus 30 days of control diet. In the third trial (90 days of β-
glucan) fish were fed the control diet or β-glucan for 90 days, β-glucan diet for 30 days 
plus 60 days of control diet and β-glucan diet for 60 days 30 more days of the control 
diet. Fish were sampled, anesthetized in clove oil (1g 10L-1 of water) and the head 
kidney was collected to assess TNF-α gene expression by qPCR RealTime. The results 
obtained in this study show that the administration of β-glucan after 30 days of feeding 
increases the expression of TNF-α in relation to its control. In the second administration 
of β-glucan after 60 days of feeding, the expression of TNF-α increased compared to 
the control. In the third administration of β-glucan after 90 days of feeding and after 60 
days of feeding with glucan plus 30 days of control diet, the expression of TNF-α 
increased in relation to the control. Therefore, this cytokine effect is an indicator of the 
activation of the immune response, which is a desired response to face challenges such 
as the emergence of pathogens. In the present study, the administration of β-glucan for 
60 days promoted an increase in the cytokines studied for up to 90 days, even after 30 
days of administration of the control diet. Other responses should be studied, but it is an 
indication of a prolonged response to this immunostimulant. 
25
 
 
Amino acids are more efficient than IGF1 in stimulating pathways of 
development and growth in pacu (Piaractus mesopotamicus) muscle cells 
 
Bruna Tereza Thomazini Zanella*, Erika Stefani Perez1, Edson Assunção Mareco2, 
Bruno Oliveira Silva Duran3, Maeli Dal-Pai-Silva1 
*Doutoranda; Instituto de Biociências de Botucatu, Universidade Estadual Paulista (UNESP); Rua Dr. 
Antônio Celso Wagner Zanin, N° 250; 18618-689 - Botucatu - SP; bruna.zanella@unesp.br; 1Instituto 
de Biociências de Botucatu, Universidade Estadual Paulista (UNESP), Botucatu, SP; 2Universidade do 
Oeste Paulista, UNOESTE, Presidente Prudente, SP; 3Instituto de Ciências Biológicas, Universidade 
Federal de Goiás (UFG), Goiânia, GO. 
 
The skeletal muscle is the main edible part of the fishes. Pro-growth factors as amino 
acids (AA) and IGF1 can enhance the growth of this tissue. In addition, these agents’ 
supplementation can be applied as a strategy to promote the recovery after muscle loss 
situations. However, the molecular mechanisms that clarify how these factors activate 
muscle gain remains unclear in fish. In this sense, in vitro studies are a helpful model to 
answer this question since it allows the evaluation of the effects of external stimulus in 
an isolated and controlled environment. Thus, this study aimed to characterize the 
transcriptional profile of pacu (Piaractus mesopotamicus) isolated muscle cells treated 
with AA or IGF1. Muscle cells were isolated from juvenile pacu (20 g) and after the 
eighth day of culture, the cells were subjected for 12 hours to a nutrient restriction media 
to decrease the gene expression to basal levels. Next, the cells were supplemented with 
AA or IGF1 for 24 hours, and a Control group was kept in the starvation situation for 
the same amount of time (CEUA: 1184). Total RNA was extracted and sequenced in 
Illumina NovaSeq 6000 system. The differentially expressed genes (DEGs) in AA and 
IGF1 groups were analyzed in the R environment applying as parameter Log2 
FoldChange 1 and -1 related to the Control group (p < 0.05). The biological processes 
enriched by the DEGs were analyzed using the Fish Enrichr database. We observed, 
respectively, 1,228 and 534 DEGs in AA and IGF1 groups. We found similar pathways 
changed by both treatments as Stat phosphorylation (GO:0007260), Pi3k positive 
regulation (GO:0014068), and AA transport (GO:0015807; GO:0015800). Despite the 
observed similarities, the AA supplementation also stimulated other processes essential 
for muscle growth, as chromosomal rearrangement (GO:0030261; GO:0000819), cell 
differentiation (GO:0045445; GO:0035914), and muscle development (GO:0048641), 
indicating a direct and immediate effect in muscle mass increase. Otherwise, the 
alterations promoted by the IGF1 treatment were restricted to its own pathway 
(GO:0043567; GO:0008286) and AA metabolism (GO:0006526; GO:0006536; 
GO:0009074). Our findings could bring an explanation of why AA can promote muscle 
growth more efficiently than IGF1 in fishes and which genes and pathways act in this 
process. Besides, the transcriptional signature of the isolated cells could provide the 
molecular characterization of the anabolic activity promoted by different stimuli in fish 
skeletal muscle. 
 
Grants: FAPESP 2018/26428-0; CAPES 88887.482392/2020-00 
26
 
 
Oregano essential oil supplementation for the catfish Lophiosilurus alexandri fed 
with high plant protein diets 
 
 
Cristiana Leonor da Silva Carneiro1*, André Luis Souza Modesto1, Rafael Costa 
Teixeira Rusth1, Roger Iván Valderrama Londoño1, Daniel Abreu Vasconcelos 
Campelo2, AntônioPolicarpo Souza Carneiro1, Jener Alexandre Sampaio Zuanon1, 
Ana Lúcia Salaro1, 
 
*Doutoranda em Biologia Animal; Universidade Federal de Viçosa ; Av. Peter Henry Rolfs, s/n, 36570-
900, Viçosa, Minas Gerais, Brasil; cristiana.carneiro@ufv.br; 1Universidade Federal de Viçosa , Viçosa, 
MG; 2Universidade Federal do Pará . 
 
Diets with high levels of plant protein ingredients can damage the intestinal mucosa and 
reduce the growth performance of aquatic organisms, especially carnivorous fish. 
Consequently, the feed additives that can improve intestinal health, digestion, and 
nutrient utilization efficiency in these diets can contribute to enhance the production of 
these species. Essential oils have been indicated as dietary additives for fish as they 
contain bioactive compounds that improve intestinal health, feed efficiency and fish 
growth. Therefore, the aim of this study was to evaluate whether dietary 
supplementation with oregano essential oil (OEO) improves growth performance and 
intestinal histomorphometry of juvenile Lophiosilurus alexandri catfish fed with a diet 
of high plant protein content. A basal diet (353.6 g kg−1 crude protein and 4464.75 g 
kg−1 crude energy) with 54% soybean meal was supplemented with four different levels 
of OEO (0.0, 1.0, 2, 0 and 4.0 g kg− 1). A quadruplicate feeding trial was carried out 
with juveniles of 15.7 ± 1.8 g and 9.5 ± 0.45 cm for 13 weeks. Regarding growth 
performance, the following variables were evaluated: survival rate, weight gain, length 
gain, specific growth rate, and viscerosomatic, visceral and hepatosomatic fat. 
Concerning intestinal histomorphometry, they were evaluated villi’s height and width, 
surface area of intestinal absorption, and thickness of the tunic muscle. According to the 
polynomial contrasts, there was no effect (P>0.05) of dietary OEO supplementation for 
growth performance and intestinal histomorphometry of fish. It was concluded that 
OEO did not improve intestinal health and growth performance of L. alexandri juveniles 
fed with a diet of high plant protein content. 
 
Key-words: feed additive, intestinal histomorphometry, pacama, soybean meal 
 
Finance Institutions: CAPS, CNPQ and FAPEMIG 
27
 
 
Antimicrobial activity of essential oils against Klebsiella pneumoniae isolated 
from diseased Nile tilapia farmed in Brazil 
 
Daiane Vaneci-Silva1*, Inácio Mateus Assane1,2, Fabiana Pilarski1 
 
*Master student, 1Laboratory of Microbiology and Parasitology of Aquatic Organisms, Aquaculture 
Center of São Paulo StateUniversity (Caunesp), Via de Acesso Prof. Carlos Donato Castellane s/n, 
1488-900 – Jaboticabal – SP, Brazil;daianevaneci@hotmail.com; 2Zambeze University (UniZambeze), 
Faculty of Agricultural Sciences, 073-02 – Ulónguè – Tete, Mozambique, 
 
 
The intensification of aquaculture has contributed significantly to the emergence of 
several fish diseases, mainly of bacterial origin. Disease outbreaks are a great threat to 
the expansion of aquaculture. Although several chemical and veterinary products are 
used both for the treatment and prevention of fish diseases in commercial aquaculture, 
there is still a lack of approved effective antimicrobials for use in food-fish species. In 
this study, we evaluated the in vitro antimicrobial activity and cytotoxicity of sixteen 
essential oils (EOs) of plants (Artemisia vulgaris, Citrus bergamia, Cedrela fissilis, 
Cymbopogon winterianus, Copaifera langsdorffii, Eucalyptus spp., Corymbia 
citriodora, Zingiber officinale, Pelargonium graveolens, Mentha piperita, Ocimum 
basilicum, Melaleuca alternifolia, Boswellia carterii, Citrus aurantium, Thymus 
vulgaris, Cananga odorata), and two commercial antimicrobials approved for use in 
aquaculture (florfenicol and oxytetracycline) against seven strains of Klebsiella 
pneumoniae isolated from diseased Nile tilapia farmed in Brazil and Nile tilapia red 
blood cells, respectively. The antimicrobial activity was determined by the 
microdilution method and cytotoxicity by haemolysis assay. Thyme (T. vulgaris) EO 
was the most active compound against K. pneumoniae, with minimum inhibitory and 
minimum bactericidal concentrations (MIC and MBS, respectively) ranging from 1.25 
µg mL-1 to 2.5 µg ml-1. Tea tree (M. alternifolia) EO presented bactericidal activity 
against all strains evaluated (MIC and MBC = 10 µg mL-1). Mugwort (A. vulgaris), 
Eucalyptus spp., bergamot (C. bergamia), ylangylang (C. odorata), frankincense (B. 
carterii), and geranium (P. graveolens) EO also showed good antimicrobial activity 
(MIC ≤ 20 µg mL-1). On the other hand, all strains were resistant to both commercial 
antimicrobials (MIC ≥ 64 µg mL-1). The main components of the most active EOs 
were thymol (53.3% - thyme EO), p-cymene (20.8% - thyme EO), 1-terpinen-4-ol 
(40.5% - tea tree), γterpinene (19.7 % - tea tree) and α-terpinene (1.6% - tea tree). 
Only tea tree EO showed haemolytic activity against Nile tilapia red blood cells. 
These results indicate that most of the EOs studied have the potential as a good 
treatment option against K. pneumoniae in Nile tilapia aquaculture. 
28
 
 
Growth and survival of Nile tilapia fed diets containing nucleotides and 
challenged against Streptococcus agalactiae 
 
Evandro Bilha Moro*, Inácio Mateus Assane1, Suzana Kotzent1, Karen Dayana Prada 
Mejia2, Bruno Luis Miani Verri1, André do Vale Oliveira1, Fabiana Pilarski1 
 
*PhD Student; Centro de Aquicultura da Unesp; Via de Acesso Professor Paulo Donato Castellane, s/n - 
Vila Industrial, 14884-900, Jaboticabal - SP; evandro.moro@unesp.br; 1Universidade Estadual Paulista 
“Júlio de Mesquita Filho”, SP, 2Universidade Nilton Lins, AM. 
 
Food consumption of fish meat has grown worldwide. To serve the sector, aquaculture 
production has been intensified. As a result, animals have been subjected to stressful 
conditions, making them susceptible to disease. This has an environmental and 
economic impact. Thus, feed supplemented with products that add animal health has 
been preferred. The inclusion of products that stimulate the immune system (prebiotics, 
probiotics, organic acids), provide better performance, balance with gastrointestinal 
microbial communities and improve the immune response. The present study aimed to 
evaluate an incorporation of the nucleotide in Nile tilapia diets and its efficacy when 
challenged against Streptococcus agalactiae. Were used two hundred and ten juvenile 
Nile tilapia with mean weight of 21.70 ± 0.54 g and mean total length of 10.74 ± 0.33 
cm. The fish were randomly distributed in 21 experimental units of 310 L, with constant 
aeration, thermostat and independent water renewal system. An isoprotein (30%) and 
isonergic (4197 kcal) diet was elaborated. Diets were divided into two groups and three 
levels of nucleotide supplementation. In group one, nucleotide was added prior to the 
extrusion process, and in group two it was added after extrusion using 
carboxymethylcellulose. Supplementation levels were 250, 500 and 1000ppm of 
nucleotide. A diet without the addition of nucleotides called control diet was maintained. 
The fish were fed three times a day for 32 days. At the end of the experimental period, 
were evaluated growth in weight and length, daily weight gain and feed conversion rate. 
Still at the end, the fish were challenged against Streptococcus agalactiae and observed 
their behavior and daily mortality for fifteen days. Data were prepared by analysis of 
variance and, when significant difference was shown (P < 0.05), by Holm-Sidak method 
was applied. Fish fed with 1,000ppm of nucleotide incorporated before extrusion 
showed higher values for final average weight, final average length and weight gain. 
The group fed with 250 ppm of nucleotide after extrusion presented a lower value for 
the average final weight, average final length and weight gain. The control group 
presented a lower value for theaverage final length. After bacterial challenge, the fish 
showed characteristic clinical signs of disease and mortality, however there was no 
difference between treatments. The supplementation of 1,000ppm of nucleotide in diet 
can favor the growth of Nile tilapia, which can be included before the extrusion process. 
However, the doses used were insufficient to increase resistance against S. agalactiae. 
29
 
 
 
 
 
 
 
 
Immunostimulation: The investiment in promoting fish health 
 
Giselle Eler* 
*Pesquisadora em Nutrição de Organismos Aquáticos; Estação de Aquicultura Almirante Paulo Moreira 
(FIPERJ); Av. das Américas; 31000-501 – Rio de Janeiro - RJ; giselleeler@gmail.com; 
An immunostimulant is a therapeutic candidate that modulates or stimulates the immune 
system of its host by instigating their defence pathways. Generally, the 
immunostimulants act by encouraging the phagocytic and bacterial killing mechanisms 
of macrophages, complement pathways, lymphocytes and innate cytotoxic cells for 
achieving enhanced immunity. The immune system is responsible to maintain the 
organism's homeostasis when invaded by foreign object or organisms. Most pathogens 
and danger particles can be recognized by immune cells through expressed pathogen or 
danger–associated molecular patterns, or missing major histocompatibility (MHC) class 
I molecules, presenting foreign non–self peptides of intracellular (through MHC class 
1-e.g. virus–infected target cells) or extracellular (through MHC class II–e.g. from 
bacteria) origin. Specialized immune cells of the innate and adaptive responses are 
involved to eliminate invaders directly or by destroying their ability to replicate. In order 
to decrease the environmental stress and thus reduce the prevalence of the disease and 
the use of chemotherapy, fish culture management techniques are designed. Diseases 
are among the primary limiting factors for the growing of aquaculture, where bacterial 
infections are responsible for heavy mortality in both wild and cultured fish. The 
majority of bacterial diseases in fish are caused by short, Gram-negative rods belonging 
to the families Enterobacteriaceae, Pseudomonadaceae or Vibrionacea. The use of 
immunostimulants, as dietary supplements, can improve the innate defense of animals 
providing resistance to pathogens during period of high stress, such as grading, 
reproduction, sea transfer and vaccination. The application of immunostimulants is 
increasing currently because of the frequent outbreaks of fish diseases in culture systems 
and hatcheries. Immunostimulants receiving major attention and claiming success in 
rendering enhanced protection in fish under experimental or practical conditions, 
independent of their mode of action. Depending on the characteristics of 
immunostimulants, these can be broadly classified into two categories namely immuno-
nutrients and immuno-additives. Immune-nutrients refer to the additional nutrients, 
either essential or non-essential, that are supplied through an exogenous source and aid 
in the functioning of the immune system against invading pathogens (e.g. functional 
amino acids, nucleotides, fatty acids, antioxidant micronutrients, vitamins and mineral 
nanoparticles). Immune-additives refer to the additional substances in feed, with known 
bioactivity and immune-boosting capability. Together with immune nutrients, these are 
intended to serve specific needs of the growing animal as quality ingredients (e.g. 
probiotics, prebiotics, synbiotics, phytobiotics and antimicrobial peptides). Dietary 
manipulation is one of the most feasible and commercially viable strategies for boosting 
“preventive-health”, by influencing the domain of immunity, through nutrition. 
30
 
 
 
Resistance of Nile Tilapia supplemented with zinc oxide nanoparticle and red 
aroeira extract against Streptococcus agalactiae 
 
Hugo Leandro dos Santos*, Amanda Silva Carvalho1, Jéssica Maria Fontes Santos1, 
Shirley Ávila Meneses1, Fabrício Sa de Santana1, Cindy Caroline Moura Santos2, 
Thays Brito Reis Santos2, Peterson Emmanuel Guimarães Paixão2, Higo Andrade 
Abe3, Rodrigo Yudi Fujimoto4 
 
*Graduate at Engenharia de pesca, Federal University of Sergipe; Av. Marechal Rondom; 49100-000- 
São Cristóvão- SE; hugoleandrobfc149@gmail.com; 1Federal University of Sergipe, SE; 2Tiradentes 
University, SE; 3Federal University of Pará, PA; 4Brazilian Agricultural Research Corporation, SE 
 
 
Nile Tilapia is the most reared fish species in the Brazil, but the intensification of its 
production have been caused diseases outbreaks mainly by bacteria as Streptococcus 
agalactiae. To control and to prevent the bacterial diseases, zinc and phytotherapics 
has been used in aquaculture. For better utilization of minerals, the nanotechnology is 
a new approach to potentialize its effects. Furthermore, the red aroeira stand out as one 
of the most popular medicinal plants due to their benefits to health. For this reason, 
this study evaluated the effect of zinc oxide nanoparticle (NPZnO) and alcohol extract 
of red aroeira (AEA) supplemented in the diet for Nile Tilapia challenged against 
Streptococcus agalactiae. Nanoparticles were synthetized by chemical method using 
dehydrated zinc acetate as oxidant agent and alcohol extract by static maceration of 
aroeira leaf into ethanol (99.8%) per 1 hour at room temperature. Basal diets (specific 
for Nile Tilapia) were formulated and added 15mg/kg of NPZnO and 100mL/kg of 
AEA. All animals received the trial diets throughout 90 days and then submitted to 
trial challenge against pathogenic bacterium S. agalactiae. The challenge consisted of 
fish fed with NPZnO, AEA, and without supplementation injected with pathogen, plus 
two control groups (positive: fish without supplementation injected with pathogen; 
negative: fish without supplementation and no injected). Each fish received an 
intraperitoneal injection containing 100 µL of S. agalactiae at concentration 1x107 
CFU/mL. This experiment occurred for 120 hours determining any possible mortality. 
All data were conducted to analysis of variance (ANOVA) followed by Tukey test 
(α=0.05). As result, the positive control and NPZnO obtained the highest mortality 
rates (46.6%) at the end of experimental period. Fish fed with AEA not presented any 
mortality. Thus, the use of red aroeira ethanol extract improve the resistance of Nile 
Tilapia against S. agalactiae. 
 
31
 
 
 
Propolis (red and green) and virgin coconut oil as immunostimulant for angelfish 
(Pterophyllum scalare) 
 
Fabricio Sa de Santana1, Jéssica Maria Fontes Santos1, Amanda Silva Carvalho1, Hugo 
Leandro dos Santos*, Shirley Avila Meneses1, Cindy Caroline Moura Santos2, Thays 
Brito Reis Santos2, Peterson Emmanuel Guimarães Paixão2, Higo Andrade Abe3, 
Rodrigo Yudi Fujimoto4. 
 
*Graduate at Engenharia de pesca, Federal University of Sergipe; Av. Marechal Rondom; 49100- 
000- São Cristóvão- SE; hugoleandrobfc149@gmail.com; 1Federal University of Sergipe, SE; 
2Tiradentes University, SE; 3Federal University of Pará; 4Brazilian Agricultural Research Corporation 
 
The angelfish (Pterophyllum scalare) is one of the most reared Amazon ornamental fish. 
However, diseases outbreaks (caused by parasite or bacterium) still are a problem for 
its rearing in captivity. Fish farmers and researchers have been searching for different 
alternatives to ensure the health for fish. Nowadays, feeding additives stand out as an 
efficient alternative to improve the immunological system. Propolis and virgin coconut 
oil are cited by its benefits in health when supplemented in the diet. Nonetheless, 
scientific reports still are missing for angelfish. For this reason, this study evaluated the 
dietary supplementation with propolis (red and green) and virgin coconut oil for 
angelfish. Therefore, alcohol extracts were prepared using 1g of propolis into10mL of 
ethanol, during 1 hour at room temperature, filtered and then sprinkled on the diet. The 
experiment occurred in completely randomized design in factorial arrangement (3x3) 
containing three levels of virgin coconut oil (0, 50 and 100% of VCO replacing Soybean 
Oil), propolis supplementation (none propolis, red and green propolis at proportion 
10g/kg) in triplicate. At the end of 90 days, nine fish per treatment had blood sampled 
for analysis of erythrogram and leukogram. There is no any statistical interaction 
between propolis (red or green) and virgin coconut oil. Separately, concentrations of 50 
and 100% of virgin coconut oil promoted high count of erythrocytes, total of leukocytes 
and lymphocyte values. Increased values of thrombocytes occurred only in the 
treatments containing red propolis or 100% virgin coconut oil. For monocyte, increasing 
levels of coconut oil in the diet, increased the number of this cell in the blood. As 
conclusion, virgin coconut oil (50 or 100%) and red propolis improve the 
immunological system. 
 
 
32
 
 
 
 
 
 
 
 
 Interleukin 10 (IL-10) is an anti-inflammatory compound which suppresses macrophages, 
pro-inflammatory cytokines, and the production of reactive species. IgM is a protein for 
antigen recognition, and also a good marker of the immune status in fish, as it is the 
universal antibody and the only component of the specific humoral immune response in 
teleost. Thus, the aim of this study was to evaluate the effect of β-glucan supplementation 
in the diet associated with vaccination on the expression of the IL-10 and IgM gene in the 
cranial kidney and spleen of tilapia (Oreochromis niloticus) against an immunological 
challenge with S. agalactiae bacteria. The experiment was conducted in a completely 
randomized design with three trials and was designed to assess the association between 
β-glucan supplementation, vaccination and a challenge with inactive bacteria. In the first 
assay (15 days β-glucan), fish were fed with either the control or the β-glucan diet for 15 
days and then sampled. In the second assay (30 days post-vaccine), fish were fed with 
either the control or the β-glucan diet for 30 days, vaccinated (AquaVac® STREP SA, 
MSD) or not-vaccinated at 15 days of feeding, and sampled at 30 days post-vaccine and, 
in the third assay (15 days post-challenge), fish were fed with either the control or the β-
glucan diet for 30 days, vaccinated or not-vaccinated at 15 days of feeding, challenged 
with inactive S. agalactiae bacteria at 30 days post-vaccine, and sampled at 15 days post-
challenge (an extra unchallenged control group was included). The results were submitted 
to analysis of variance (ANOVA) and means compared by Tukey test (5%). At 30 days 
post-vaccine, a significant increase was observed in HK of the vaccinated + β-glucan 
group compared other treatments. At 15 days post-challenge, there was increased 
expression in the HK in all treatments compared to SP, except in the challenged control 
group, as it increased significantly in the β-glucan group, followed by the vaccinated + β-
glucan group. Conversely to the above profiles, the IgM gene expression significantly 
increased in SP. IgM expression increased in SP after 15 days of β-glucan compared to 
HK. At 30 days post-vaccine, there was increased IgM expression in SP in all groups 
compared to control but not in HK. At 15 days post-challenge, there was increased 
expression in HK in the control group but not in the SP. Thus, it is concluded that there 
is a beneficial interaction between the administration of β-glucan and vaccination in 
tilapia. 
β-glucan stimulates the expression of interleukin 10 (IL-10) and immunoglobulin 
M (IgM) to prevent pathogens entrance in vaccinated Nile tilapia 
 
Ingrid Camargo dos Reis1, Camino Fierro-Castro 2, Raphael Farias Mathias1, Jaqueline 
Dalbello Biller 1 
 
¹ Faculdade de Ciências Agrárias e Tecnológicas, FCAT-UNESP, Campus de Dracena, Brasil. 
ingrid.c.reis@unesp.br; ² University of Leon. Faculty of C.C. Biológica y Ambientales. Campus of 
Vegazana, Leon, Spain 
33
 
 
 
 
 
 
 
 
 Recently, there are an increase in studies to understand the immune system mechanism 
in from of pathogen challenges to create measures to mitigate economic loss of diseases. 
Thus, some genes have been extensively studied due to their importance for immune 
system function, among them, 70kb heat shock protein (HSP70). HSPs are called 
chaperones, because they help to properly assemble proteins during synthesis or repairing 
proteins. Thus, the aim of this study was to evaluate the effect of β-glucan 
supplementation in the diet associated with vaccination on the expression of the IgM and 
IL-10 gene in the cranial kidney and spleen of tilapia (Oreochromis niloticus) against an 
immunological challenge with S. agalactiae bacteria. The experiment was conducted in 
a completely randomized design with three trials and was designed to assess the 
association between β-glucan supplementation, vaccination and a challenge with inactive 
bacteria. In the first assay (15 days β-glucan), fish were fed with either the control or the 
β-glucan diet for 15 days and then sampled. In the second assay (30 days post-vaccine), 
fish were fed with either the control or the β-glucan diet for 30 days, vaccinated 
(AquaVac® STREP SA, MSD) or not-vaccinated at 15 days of feeding, and sampled at 
30 days post-vaccine and, in the third assay (15 days post-challenge), fish were fed with 
either the control or the β-glucan diet for 30 days, vaccinated or not-vaccinated at 15 days 
of feeding, challenged with inactive S. agalactiae bacteria at 30 days post-vaccine, and 
sampled at 15 days post-challenge (an extra unchallenged control group was included). 
The results were submitted to analysis of variance (ANOVA) and means compared by 
Tukey test (5%). The HPS70 expression in the HK increased in all treatments compared 
to the SP, except in the control whereas suppression was observed in SP after treatments. 
At 15 days, HPS70 expression increased in the HK in the β-glucan group but not in SP 
compared to control. At 30 days post-vaccine, HSP70 increased in the HK of the 
vaccinated + β-glucan group. At 15 days post-challenge, HSP70 increased in the HK in 
the β-glucan group compared the other groups, except for the vaccinated control group, 
but no changes were observed in SP. The β-glucan and vaccination caused enough 
stimulus for HPS70 to be expressed in the head kidney of the fish in the present study. 
Thus, it is concluded that there is a beneficial interaction between the administration of 
β-glucan and vaccination in tilapia. 
β-glucan potentiates the expression of HsP70 gene in vaccinated Nile tilapia 
Ingrid Camargo dos Reis1*, Camino Fierro-Castro2, Thais Cristina de Oliveira1 
Jaqueline Dalbello Biller1 
 
¹ Faculdade de Ciências Agrárias e Tecnológicas, FCAT-UNESP, Campus de Dracena, Brasil. 
ingrid.c.reis@unesp.br; ² University of Leon. Faculty of C.C. Biológica y Ambientales. Campus of 
Vegazana, Leon, Spain 
34
 
 
 
 
 
 
 
 
 Currently there are an improvement in studies with immunostimulants, among them 
the β-glucan, which has a powerful action in the protection of fish against many 
diseases. β-glucans are glucose polymers of fungal walls, such as yeast Saccharomyces 
cerevisiae, with a potent action on the production of immunological cytokines. IL-1β 
and IL-8 are pro-inflammatory cytokines involved in the immune response in a wide 
range of diseases. Thus, the aim of the present study was to evaluate the β-glucan 
effects on the immune system IL-1β and IL-8 cytokines gene expression. The 
experiment was conducted in a completely randomized design with three trials and 
was designed to assess theβ-glucan effect over time. For this purpose, tilapias were 
fed with a control and an experimental diet with 10g kg-1 of β-glucan for 30, 60 and 
90 days. In the first assay (30 days β-glucan), fish were fed with either the control or 
the β-glucan diet for 30 days and then sampled. In the second assay (60 days β-glucan), 
fish were fed with either the control or the β-glucan diet for 60 days and β-glucan diet 
for 30 days plus 30 day of control diet. In the third assay (90 days β-glucan) fish were 
fed with either the control or the β-glucan diet for 90 days, β-glucan diet for 30 days 
plus 60 day of control diet, and β-glucan diet for 60 days plus 30 day of control diet. 
Fish were sampled, anesthetized in clove oil (1g 10L-1 of water), and head kidney was 
collected to evaluate the IL-1β and IL-8 gene expression by qPCR RealTime. In the 
first assay, after 30 days of β-glucan feeding, IL-8 expression increased compared to 
its control. In the second assay, after 60 days of β-glucan feeding, IL-1β and IL-8 
expression increased compared to their control. In the third assay, after 90 days of β-
glucan feeding, and after 60 days of glucan feeding plus 30 days of control diet, IL-1β 
and IL-8 expression increased compared to their control. The increase in cytokines is 
a sign of the immune response activation, which is a desired response to face 
challenges such as disease outbreaks. In the present study, 60 days β-glucan 
administration promoted an increase in the studied cytokines for up to 90 days, even 
after 30 days of control diet administration. Other responses should be studied but it is 
an indication of a prolonged response of this immunostimulant. 
Grants: The São Paulo Research Foundation - FAPESP – Process number: 
2012/22016-3. 
β-glucan administration for 60 days stimulates the IL-1β and IL-8 gene 
expression for up to 90 days in Nile tilapia 
Jaqueline Dalbello Biller1*, Ingrid Camargo dos Reis1, Laura Cristina de Paula1, Basia 
Schlichting Moromizato1 
 
¹ Faculdade de Ciências Agrárias e Tecnológicas, FCAT-UNESP, Campus de Dracena, Brasil. 
jaqueline.biller@unesp.br. 
35
 
 
 
 
 
Microbiome of Nile tilapia juvenile fed diets with xylanase and β- glucanase 
 
João Antônio Galiotto Miranda*, Johnny Martins de Brito1, Paola Aparecida 
Paulovski Panaczevicz1, Allan Vinicius Urbich1, Angelo Machado Soltes Filho2, Ana 
Flávia Moreira2, Valéria Rosseto Barriviera Furuya3, Wilson Massamitu Furuya3 
*Graduação em Zootecnia, Universidade Estadual de Ponta Grossa (UEPG) - Ponta Grossa - PR; 
joaoantonio99live@hotmail.com; 1 Programa de Pós-Graduação, Universidade Estadual de Maringá; 2 
Graduação em Zootecnia, UEPG; 3 Departamento de Zootecnia, UEPG. 
 
Diets with vegetable source ingredients have non-starch polysaccharides (NSP), those 
are antinutricional factors that restringe the use of those ingredients and modify the 
intestinal microbiome, developing the anaerobic microbiome, considered unwanted. 
The carbohydrases have an effect on the quantity and type of the present microbiome. 
This way, this paper aimed to evaluate the effects of xylanase-β-glucanase (XB) on the 
intestinal microbiome of Nile tilapia juveline. For that, 136 Nile tilapia juveline (12 ± 
1,4 g) were distributed with completely randomized design in eight aquariums, being 
17 fishes by experimental unit, with two treatments, diets exclusively of vegetable 
source with and without addition of XB (0,2 g kg-1) and four repeats, by 90 days. It was 
added on top on the diet 0,20 g kg-1 of liquid XB (Natugrain®, Basf, Ludwigshafen am 
Rhein, Germain), enzymatic complex made of endo-1,4-β-xylanase 5600 TXU g-1 and 
endo-1,4-β-glucanase 2500 TGU g-1. The intestinal content DNA analysis was realized 
with ZR Fecal DNA MiniPrep® kit, followed by spectrophotometry and ampliation to 
build a metagenomic library. The sequences were classified in genus by the knowledge 
of operacional taxonomic units (OTUs). To the statistical analysis of the microbiome 
was used the Welch test (P < 0,05), followed by the correction test of Bonferroni, the 
biodiversity avarage was compared by the OTUs number and the Kruskal Wallis test (P 
< 0,05) The statistical program used was STAMP (Statistical Analysis of Metagenomic 
Profiles). There were identified 243 taxa, being the majority Barnesiella (20 %), 
followed by Bacteroides (16 %), Alistipes (9 %) and Faecalibacterium (4 %). It was 
identified 25 different genus (P > 0,05) among the treatments, knowing that the fished 
fed with the diet with XB supplementation, when compared to fishes that recieved the 
diet without XB, showed increase of the genus Basnesiella (31,878 % vs. 22,435), 
Bacteroides (20,530 % vs. 16,351 %), Ruminococcus torques (4,892 % vs. 1,963 %), 
Peptoclostridium (2,372 % vs. 0,126 %), Anaerofilum (1,287 % vs. 0,274 %), 
Butyricicoccus (1,167 % vs. 0,738 %) and Parasutterella (1,121 % vs. 0,001). Diets 
without XB addition showed 1900 unique OTUs, fishes fed with XB addition showed 
2355 unique OTUs. The core microbiota between the treatments was 1897 OTUs, some 
of the identified genus were Barnesiella, Bacteroides, Faecalibacterium and 
Butyricicoccus It was concluded that the XB supplementation modulated the intestinal 
microbiome, reducing or increasing the presence of some bacteria, in majority the 
Barnesiella genus. The increase in quantity and variety of the population takes to a 
higher absorption and digestion rate of the nutrients by the fish, this take the diet more 
efficient for the fish, reducing producing cost and increasing profit. 
36
 
 
 
 
 
 
 
 
 β-glucans are glucose polymers found in the fungi and yeast walls, and show potent 
action on immune system. One of the most important defense mechanisms of the innate 
immune system is phagocytosis, and can be assessed by the leukocyte respiratory burst 
activity in the blood. There are many studies with β-glucan, but little is known about 
the effects of interactions with vaccines on phagocytosis. Thus, the aim of this study 
was to evaluate the effect of β-glucan supplementation in the diet associated with 
vaccination on the blood leukocytes respiratory burst activity (BURST), besides to 
standardize the microplate method in tilapia (Oreochromis niloticus). The experiment 
was conducted in a completely randomized design, and tilapias were divided in four 
treatments: fed with a control diet (C), fed with a control diet and vaccinated (C+V), 
fed an experimental diet with 1g kg-1 of β-glucan for 15 days (BG), and fed an 
experimental diet with 1g kg-1 of β-glucan for 15 days and vaccinated (BG+V). Fish 
were vaccinated (AquaVac® STREP SA, MSD) on the 15th day of the experiment. 
After 30 days of experiment (15 days after vaccination) fish were sampled, 
anesthetized in clove oil (1g 10L-1 of water), and blood was collected to evaluate the 
BURST by microplate method. The results were submitted to analysis of variance 
(ANOVA) and means compared by Tukey test (5%). After 30 days of experiment, β-
glucan promoted an increase in BURST compared to the β-glucan fed and vaccinated 
group. The immunostimulant induced an increase in BURST even after 15 days of 
feeding with the control diet. Other studies should be carried out in other post-vaccine 
periods to evaluate these responses over time. Thus, it is concluded that the β-glucan 
may improve phagocytosis activity, however there is a negative interaction between 
the administration of β-glucan and vaccination in tilapia, which could be an 
immunosuppression indication. 
Grants: The São Paulo Research Foundation - FAPESP – Process number: 
2012/22016-3. 
 
β-glucan effects on blood leukocyte respiratory burst activity 
Laura Cristina de Paula1 Ingrid Camargo dos Reis1*, Nathália Gabrielle Aranibar 
Passos1, Jaqueline Dalbello Biller1 
 
¹ Faculdade de Ciências Agrárias e Tecnológicas, FCAT-UNESP,