Sousa ML TCC Rcla

ECOLOGIA
MARIANA LOPES DE SOUSA
TOXICIDADE A Saccharomyces cerevisiae E

ECOTOXICIDADE DE EFLUENTE CONTENDO
CORANTE REMAZOL VERMELHO BRILHANTE
TRATADO PELO PROCESSO FOTOELETROLÍTICO
Rio Claro
2010
MARIANA LOPES DE SOUSA
TOXICIDADE A Saccharomyces cerevisiae E ECOTOXICIDADE

DE EFLUENTE CONTENDO CORANTE REMAZOL
VERMELHO BRILHANTE TRATADO PELO PROCESSO
FOTOELETROLÍTICO
Orientador: Ederio Dino Bidoia
Co-orientador: Peterson Bueno de Moraes
Trabalho de Conclusão de Curso apresentado ao

Instituto de Biociências da Universidade Estadual
Paulista “Júlio de Mesquita Filho” - Campus de Rio
Claro, para obtenção do grau de Ecóloga
Rio Claro
2010
Dedicado aos meus pais, Jesus e Rosa
e a minha avó Carmela, pelo amor e criação.
Ao meu namorado e melhor amigo,
Rodrigo pelo apoio incondicional
aos meus objetivos durante esse período.
AGRADECIMENTOS
Ao Profº Ederio, pela orientação, paciência (principalmente) e apoio desde o segundo ano de
graduação, que foi fundamental para a realização deste trabalho;
Aos técnicos: Fátima, Luiza e principalmente ao Roberto, por me ajudarem nas situações mais
diversas possíveis durante meus experimentos, sempre que precisei;
À Profª Dejanira, pela ajuda com os (longos) ensaios com colunas de Winogradsky e testes
com S. cerevisiae;
À minha classe, a XXXI Turma do curso de Ecologia, pelas amizades, conversas, risadas,
festas, campos, momentos, que fizeram parte da minha formação;
Aos meus amigos, tanto da minha cidade natal, quanto de Rio Claro, mesmo os amigos com
os quais já não mantenho mais contato, mas agradeço a todos por tocarem minha vida de
algum modo e ajudarem a tecer a colcha de retalhos que sou hoje, e que me fez chegar até
aqui;
Á minha família, meus pais Jesus e Rosa, minha avó Carmela, e meus padrinhos José Luiz e
Geralda, por me incentivarem, e me apoiarem sempre á sua maneira, sempre o tanto quanto
possível;
Ao meu namorado, companheiro, amigo, Rodrigo, que nestes 4 anos esteve ao meu lado em
todos os meus planos, desejos, devaneios, risadas, mesmo quando estive doente, mesmo
quando me curei.
Obrigada !!!
As estrelas brilham, mas um dia se apagarão...
Tudo morre...
A terra, o sol, a via láctea, e até mesmo todo este universo não é exceção!
Comparado a isto,
a vida do homem é tão breve e fugidia quanto um piscar de um olho...
Neste curto instante,
os homens nascem, riem, choram,
Lutam, sofrem,
Festejam, lamentam,
odeiam pessoas e amam outras!
Tudo é transitório...
Masami Kurumada
SUMÁRIO
1. INTRODUÇÃO........................................................................................................... 7
2. OBJETIVOS.................................................................................................................9
2.1. Objetivo geral..................................................................................................9
2.2. Objetivos específicos.......................................................................................9
3. REVISÃO BIBLIOGRÁFICA ..................................................................................11
3.1. Indústrias têxteis.............................................................................................11
3.2. Eletrooxidação................................................................................................12
3.3. O corante remazol vermelho brilhante e os corantes reativos .......................13
3.4. Saccharomyces cerevisiae..............................................................................15
3.5. Coluna de Winograsky...................................................................................16
3.6. Algas e fitoplâncton.......................................................................................17
3. MATERIAL E MÉTODOS ......................................................................................20
4.1. Material..........................................................................................................20
4.1.1. Sistema fotoeletrolítico...................................................................20
4.1.2. Organismos teste............................................................................23
4.1.3. Solo e água utilizados nas Colunas de Winogradsky.....................23
4.1.4. Reagentes e Equipamentos.............................................................23
4.2. Métodos.........................................................................................................24
4.2.1. Preparo de efluente simulado.........................................................24
4.2.2. Tratamento fotoeletrolítico e retirada das amostras durante o
processo e verificação da absorbância e concentração...................26
4.2.3. Medidas de pH e condutividade.....................................................27
4.2.4. Ensaios de toxicidade com S. cerevisiae .......................................27
4.2.4.1. Preparo da suspensão de S. cerevisiae.............................27
4.2.4.2. Preparo dos tubos de ensaio.............................................28
4.2.4.3. Contagem de células de S. cerevisiae..............................28
4.2.5. Demanda química de oxigênio.......................................................29
4.2.5.1. Preparo dos tubos de DQO...............................................30
4.2.5.2. Leitura dos resultados no espectrofotômetro Hach..........31
4.2.6. Montagem da coluna de Winogradsky...........................................31
4.2.7. Análise do crescimento e da população fotossintetizante..............34
5. RESULTADOS E DISCUSSÕES ............................................................................36
5.1. Remoção da cor.............................................................................................36
5.1.1. Experimentos 1 e 2.........................................................................36
5.1.2. Experimentos 3 e 4.........................................................................39
5.1.3. Experimentos 5 e 6.........................................................................43
5.1.4. Experimento 7................................................................................47
5.1.5. Experimento 8.................................................................................48
5.2. Condutividade e pH.......................................................................................49
5.2.1. Experimentos 1 e 2.........................................................................49
5.2.2.Experimentos 3 e 4..........................................................................51
5.2.3. Experimentos 5 e 6.........................................................................54
5.2.4. Experimento 7.................................................................................57
5.2.5. Experimento 8................................................................................58
5.3. Ensaios de toxicidade com S. cerevisiae.......................................................60
5.3.1. Experimentos 1 e 2.........................................................................60
5.3.2. Experimentos 3 e 4.........................................................................64
5.3.3. Experimentos 5 e 6.........................................................................66
5.3.4. Experimento 7................................................................................68
5.3.5. Experimento 8................................................................................69
5.4. Demanda química de oxigênio......................................................................70
5.4.1. Experimentos 1 e 2.........................................................................70
5.5. Análise do crescimento na coluna de Winogradsky......................................72
5.5.1. Experimento com efluente simulado do experimento 1.................72
5.6. Avaliação da população fitoplantônica.........................................................94
5.6.3. Experimento com efluente simulado do experimento 7................100
6. CONCLUSÕES ........................................................................................................103
7. REFERÊNCIAS BIBLIOGRÁFICAS....................................................................105
7
1. INTRODUÇÃO
A preocupação com a qualidade da água é um assunto de importância institucional, social,
e política. Entretanto, a cada dia os mananciais se encontram mais poluídos por diversos detritos
despejados pela sociedade que na falta de tratamento adequado representam um sério risco sócio-
ambiental (ABREU, et al., 2008, p 160). Estima-se que no último século o consumo de água
duplicou em relação ao aumento populacional, e também que a área coberta por água doce
diminuiu pela metade (POLLI et al., 2009, p. 530).
Além do esgoto doméstico que em grande parte do país e do mundo ainda não recebe
tratamento adequado, ainda existe o grande problema da poluição causada pela indústria. Todo
dia milhares de litros de água são contaminados com diversos tipos de resíduos de natureza
orgânica e inorgânica, através dos esgotos sanitários, contendo diversos tipos de poluentes como
tanino (efluente de curtume), corantes têxteis e efluentes da indústria de papel (COMPANHIA
DE TECNOLOGIA E SANEAMENTO AMBIENTAL, 2008).
A indústria têxtil é responsável por um grande volume de água contaminada emitida
diariamente, principalmente por corantes nos processos de diluição e beneficiamento
(GEORGIOU et al., 2001, p. 69).

8
A água proveniente deste descarte diário é imprópria tanto para consumo humano, quanto para a
sobrevivência das espécies que se encontram nos rios. Entre os desequilíbrios ambientais
causados por estes efluentes pode-se citar o impedimento dos processos de fotossíntese devido à
turbidez da água (CATANHO et al., 2006-a, p. 983), danos a estruturas como guelras e
brânquias, perturbação em locais de desova e alterações na solubilidade dos gases (ZANONI e
CARNEIRO, 2001, p. 62).
Nestes efluentes também é possível encontrar toxinas que se acumulam na cadeia trófica
(ZANONI e CARNEIRO, 2001, p. 62), além de muitas vezes conterem substâncias tóxicas
mutagênicas e carcinogênicas e metais pesados em quantidades significativas (AKYOL &
BARAMOGLU, 2005, p. 241). Apenas o número de corantes carcinogênicos pode chegar a 3000
(DELLAMATRICE, 2005, p. 21; CATANHO et al., 2006-b, p. 193), sendo que o total de
corantes têxteis, segundo o Colour Índex (catálogo da Society of Dyers and Colourists) é de cerca
de 8000 (ZANONI e CARNEIRO, 2001, p. 60). Por isso é necessário o desenvolvimento de
processos para degradação ou retirada destes compostos. Tendo em vista que a preocupação com
a qualidade da água é cada vez maior, e o equilíbrio homem-natureza se encontra cada vez mais
delicado.
Neste trabalho foi analisada a degradação de efluente têxtil simulado contendo o Corante
Remazol Vermelho brilhante pelo processo fotoeletrolítico. Também, foi analisada a degradação
mediante diferentes parâmetros de tratamento (variação de corrente, eletrólitos, pH), e feitos os
estudos toxicológicos utilizando-se Saccharomycces cerevisiae como organismo teste e
ecotoxicológico da comunidade fitoplantônica exposta ao efluente tratado através de colunas de
Winogradsky.
9
2. OBJETIVO
2.1. Objetivo geral
Degradação de efluente simulado contendo o corante têxtil Remazol Vermelho Brilhante
pelo processo fotoeletrolítico.
2.2. Objetivos Específicos
Análises dos diversos parâmetros da degradação como: pH, condutividade, absorbância,
DQO e ensaios toxicológicos utilizando-se Saccharomyces cerevisiae como organismo teste.
Verificar a influência da corrente aplicada durante o tratamento, a concentração de cloreto
de sódio usada e do pH inicial na eficiência do tratamento fotoeletrolítico.

10
Analisar a diferença de experimentos tratados pelo processo fotoeletrolítico num sistema

único em fluxo, e por etapas, ou seja, o tratamento fotolitico realizado primeiramente de forma
separada do sistema eletrolítico em fluxo.
Monitoramento ecotoxicológico do efluente simulado e sem tratamento utilizando

Colunas de Winogradsky de diferentes tamanhos para análise da comunidade fitoplantônica
formada sob diferentes condições ambientais.
11
3. REVISÃO BIBLIOGRÁFICA
3.1. Indústrias têxteis
A indústria têxtil é de grande peso na economia mundial, porém ainda existe uma lacuna
no que diz respeito à responsabilidade ambiental. Em geral no Brasil, onde a indústria têxtil tem
uma grande participação e há regiões que sofrem com problemas de escassez de água, num
estudo realizado por Abreu et al. (2008, p. 170), as empresas investem no tratamento de resíduos
não pela preocupação ambiental e social, mas sim para evitar multas e obter subsídios.
Durante o beneficiamento dos tecidos são usadas diversas substâncias inorgânicas que ao
serem lançadas como parte do efluente causa modificações físico-químicas na água como (AL-
DEGS et al., 2000, p. 927).
A coloração característica dos resíduos têxteis é resultado da etapa de tingimento e é
muito problemática do ponto de vista ambiental pela coloração intensa relacionada ao descarte de
corantes não fixados ao tecido (AL-DEGS et al., 2000, p. 927; CATANHO et al., 2006-b, p. 193).
12
Existem vários tipos de tratamentos para degradar os corantes têxteis provindos dos
efluentes industriais, por tratamentos biológicos podemos citar os processos que envolvam
fungos e bactérias na biodegradação e biosorção desses compostos presentes no efluente (AKSU,
2000, p. 81; BEYDILLI, 2000, p. 699; BERTAZZOLI & PELEGRINI 2002, p. 479). Como
alternativa aos processos biológicos existem os processos físico-químicos, nos quais estão os
processos eletrolíticos (PELLEGRINI et al., 1999, p. 83-90), oxidações usando UV (GEORGIOU
et al., 2001, p. 69-78; AKYOL & BARAMOGLU, 2005, p. 241-246), peróxido de hidrogênio,
sistema ferroso (ROBINSON et al., 2000, p. 248-249) e o próprio sistema fotoeletrolítico.
Porém, esses processos não destroem a molécula do corante, podendo apenas transformá-
la, sendo assim, presente no efluente sob outra forma (SAUER et al., 2002, p. 147; ZANONI e
CARNEIRO, 2001, p. 62).
Cada um dos tipos de tratamentos tem suas vantagens e desvantagens e procura-se
atualmente descobrir um processo de baixo custo e que seja abrangente em relação aos
compostos degradados, pois os efluentes lançados normalmente possuem diversos tipos de
corantes, detergentes, normalizantes, umectantes, tornando o efluente têxtil uma matriz de
complexa degradação, devido a especificidade de cada substância contida (ZANONI e
CARNEIRO, 2001, p. 62).
3.2. Eletrooxidação
Segundo Harris (2001, p. 379) a eletrólise pode ser caracterizada como uma reação
conduzida em sua direção não espontânea pela imposição de uma corrente elétrica. Pode-se
analisar o mecanismo oxidativo produzido no tratamento fotoeletrolítico como direto e indireto.
Na oxidação direta ocorre a degradação por transferência de carga na superfície do eletrodo, sem
interferências de outros elementos (DENG & ENGLEHARDT, 2007, p. 231). Já a oxidação
indireta ocorre de maneira relativamente rápida devido à adição de um intermediário na solução
que funciona como um catalisador, no caso do processo fotoeletrolítico utilizado, o cloreto de
sódio que produz cloro durante a eletrólise (DENG & ENGLEHARDT, 2007, p. 234).
13
A estabilidade eletroquímica e física do eletrodo torna-se um fator importante devido ao

fato dos processos já descritos só ocorrerem na presença de altos potenciais anódicos, porém isso
é alcançado devido às características do eletrodo utilizado (SAUER, 2002, p. 148). Um fator que
diminui a eficiência do processo é a recombinação dos elétrons liberados com lacunas h+, antes
que haja interação com o corante, porém a irradiação UV junto com a aplicação do potencial
anódico força a separação das espécies carregadas eliminando o problema (CATANHO et al.,
2006-a, p. 984).
3.3. O corante remazol vermelho brilhante e os corantes reativos
O Remazol vermelho brilhante, de fórmula C19H10Cl2N6Na2O7S2 (Figura 1), o qual foi

utilizado neste trabalho, é muito utilizado na indústria têxtil para o tingimento de seda, lã,
algodão e também em cartuchos de impressora (BEYDILLI et al., 2000, p. 698; CATANHO et
al., 2006-a, p. 983).
Figura 1: Formula estrutural do corante remazol vermelho brilhante.

14
Ele possui uma aderência grande às fibras de celulose, reagindo com o tecido e formando
uma ligação covalente entre as moléculas do corante e da fibra (AL-DEGS et al., 2000, p. 927),
uma característica forte dos corantes vinil sulfonados (de grupo cromóforo azo N=N), como pode
ser observado na Figura 2.
.
Figura 2: Tingimento de tecido por corante reativo (ZANONI e CARNEIRO, 2001, p. 62).
Corantes reativos vinil sulfonados (de grupo cromóforo azo N=N) representam cerca de
60% dos corantes usados em indústria têxtil (CATANHO et al., 2006-a, p. 983). Altamente
solúveis em água, em geral, têm um descarte (porcentagem do corante que não se fixa as fibras)
bastante alto, podendo chegar a até 50% (AL-DEGS et al., 2000, p. 928), sendo que, quanto
maior a concentração do corante, mais difícil é a remoção deste da água (KORBAHTI, 2007, p.
285). Devido à estrutura dos corantes reativos, que possuem o grupo cromóforo azo, os corantes
são formados por ligações fortes o suficiente para não serem rompidas por processos biológicos,
que apresentam resultados ineficientes com esse tipo de efluente, principalmente, em relação à
remoção da cor (ARAÚJO & YOKOYAMA, 2006, p. 11; AL-DEGS et al., 2000, p. 928).
Portanto, como alternativa vem sendo usados os processos físico-quimicos, como os processos
eletrolítico e fotoeletrolítico, porém estes apesar de apresentarem resultados satisfatórios ainda
são raramente usados industrialmente (ZANONI e CARNEIRO, 2001, p. 63).
15
3.4. Saccharomyces cerevisiae
Saccharomycces cerevisiae é uma levedura, um fungo unicelular normalmente esférico ou

oval, não possui filamentos e se reproduz por meio de brotamento. Portanto, as células mais
novas brotam da célula mãe, são menores e possuem parede menos resistentes, sendo mais
vulneráveis a agentes tóxicos.
Segundo Tortora et al. (2005, p. 322) a célula mãe forma uma protuberância, que após
divisão do núcleo, um dos novos núcleos gerados migra para essa protuberância, então a célula se
separa. Esse número pode chegar a até 24 células por levedura.
A S. cerevisiae é não patogênica, ou seja, inofensiva para o ser humano, e é utilizada
desde milênios para a fermentação (Figura 3), sendo vendida como fermento biológico para o
preparo de pães, usada na fermentação de vinho e para produzir álcool.
(a) (b) (c)
Figura 3: (a) S. cerevisiae vista por microscópio eletrônico de varredura, (b) vinho sendo
fermentado e (c) fermento biológico em tabletes.
Devido a seu baixo custo, fácil manuseio e biologia conhecida, é possível utilizá-la como
organismo teste em ensaios toxicológicos, como já realizados por Regis e Bidóia (2001, p. 53-
60).
16
3.5. Coluna de Winogradsky
Inventada por Sergei Nikolaievich Winogradsky em 1880, a coluna forma um micro

ecossistema fechado dentro de um recipiente de vidro (Figura 4), composto por solo (retirado
tanto de área seca, ou do fundo de um rio), água e nutrientes. Dentro desse micro ecossistema é
possível acompanhar o aparecimento dos microrganismos, sua distribuição ao longo da coluna e a
partir daí inferir o papel de cada um deles na microbiota do corpo d’água (PERÉZ, 2008, p. 373).
Figura 4: Esquematização da coluna de Winogradsky.
Como pode ser observada na Figura 4, cada zona da coluna é preferencial para
determinado organismo, principalmente populações de fitoplâcton, e entendendo-se a função
deste na biota podem-se inferir os efeitos que uma possível contaminação causaria no
ecossistema aquático. Podem-se utilizar os resultados da coluna de Winogradsky em diversos
17
tipos de estudos ambientais, vinculados ou não a efluentes lançados pela indústria, no caso deste
trabalho foi o efluente tratado pelo processo fotoeletrolítico.
3.6. Algas e fitoplâncton
As algas são organismos de natureza variada e ampla distribuição, estando presentes em

praticamente todos os corpos d’água, constituindo inúmeras espécies com importante papel na
produção de matéria orgânica dos ecossistemas. Segundo Bicudo & Menezes (2006, p. 1), algas
são talófitos e protistas clorofilados, de morfologia, ecologia e comportamento tão diferentes
entre si, o que torna sua identificação dentro de um grupo praticamente impossível.
O grupo conhecido como algas, ao contrario do que se pensa, abriga espécies tanto do
reino Plantae como Metazoa e Bacteria (BICUDO & MENEZES, 2006, p.1; BRUSCA &
BRUSCA, 2003, p. 27). Quanto ao tamanho podem medir desde milímetros até metros e podem
ser coloniais ou um único individuo. Estão divididas em 4 grandes grupos: Cianobactérias (algas
azuis), Chlorophyta (algas verdes), Phaeophyta (algas pardas) e Rhodophyta (algas vermelhas).
As cianobactérias fazem parte do reino Bactéria, e crescem em sua maioria na água doce,
mas também ocorrem gêneros marinhos, sendo consideradas cosmopolitas (PARRA & BICUDO,
1995, p. 44).
Algas verdes também estão presentes em sua maioria na água doce, mas também no solo e
troncos na forma de líquens, e podem se tratar de macro ou micro algas, sendo unicelulares ou
coloniais conforme a Figura 5b (BRUSCA & BRUSCA, 2003, p. 27-32).
Quanto às algas pardas, estas são a maioria proveniente de água marinha fria, salvo raras
exceções, não apresentam formas unicelulares e coloniais (Figura 5c).
Algas vermelhas são unicelulares e filamentosas, muito usadas na indústria como
emulsificante (agar), também fazem parte da estrutura formadora de recifes de corais conforme
Figura 5d (RAVEN et al., 2007, p. 366).
18
(a) (b)
(c) (d)
Figura 5: Algas (a) Cianobactéria – Cylindrospermun sp.; (b) Alga Verde – Pandorina
sp.; (c) Alga Parda – Laminaria sp. e (d) Alga vermelha – Gelidium sp.
Devido a sua importância em relação a produção primária, o fitoplâncton é considerado

um importante indicador de ação antrópica nos ambientes aquáticos (SILVA et al., 2009, p. 355),
sendo até 50% mais sensíveis aos poluentes que outros organismos-testes comumente usados
(KLAINE e LEWIS, 1995, p. 163). Apesar de sua grande importância ecológica, as microalgas
que formam o fitoplâncton, especialmente as cianobactérias, estão relacionadas com o fenômeno
da eutrofização (MATSUZAKI et al., 2004, p. 680). Este fenômeno ocorre quando o ambiente
aquático passa por um rápido enriquecimento, aumentando a concentração de nutrientes (o que
pode ser um fenômeno natural ou artificial, devido à ação antrópica), principalmente nitratos e
19
fosfatos (FELLENBERG, 1980, p. 25). Isto causa o rápido crescimento das algas, que,
conseqüentemente, contaminam a água com fitoplancto-toxinas que podem causar problemas
hepáticos, neurológicos, paralisias, distúrbios gastrointestinais e até morte (MATSUZAKI et al.,
2004, p. 680). Outra conseqüência da eutrofização é a desoxigenação da coluna d’água, que torna
o ambiente impróprio para sobrevivência da fauna aquática e para consumo (TUNDISI e
MATSUMURA-TUNDISI, 1992, p. 1).

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ECOLOGIA

MARIANA LOPES DE SOUSA

TOXICIDADE A Saccharomyces cerevisiae E

TOXICIDADE A Saccharomyces cerevisiae E ECOTOXICIDADE

Orientador: Ederio Dino Bidoia

Co-orientador: Peterson Bueno de Moraes

Trabalho de Conclusão de Curso apresentado ao

A preocupação com a qualidade da água é um assunto de importância institucional, social,

diminuiu pela metade (POLLI et al., 2009, p. 530).

tanino (efluente de curtume), corantes têxteis e efluentes da indústria de papel (COMPANHIA

DE TECNOLOGIA E SANEAMENTO AMBIENTAL, 2008).

A indústria têxtil é responsável por um grande volume de água contaminada emitida

diariamente, principalmente por corantes nos processos de diluição e beneficiamento

(GEORGIOU et al., 2001, p. 69).

2.1. Objetivo geral

Degradação de efluente simulado contendo o corante têxtil Remazol Vermelho Brilhante

pelo processo fotoeletrolítico.

2.2. Objetivos Específicos

Análises dos diversos parâmetros da degradação como: pH, condutividade, absorbância,

DQO e ensaios toxicológicos utilizando-se Saccharomyces cerevisiae como organismo teste.

Verificar a influência da corrente aplicada durante o tratamento, a concentração de cloreto

de sódio usada e do pH inicial na eficiência do tratamento fotoeletrolítico.

Analisar a diferença de experimentos tratados pelo processo fotoeletrolítico num sistema

Monitoramento ecotoxicológico do efluente simulado e sem tratamento utilizando

3.1. Indústrias têxteis

DEGS et al., 2000, p. 927).

A coloração característica dos resíduos têxteis é resultado da etapa de tingimento e é

A estabilidade eletroquímica e física do eletrodo torna-se um fator importante devido ao

3.3. O corante remazol vermelho brilhante e os corantes reativos

O Remazol vermelho brilhante, de fórmula C19H10Cl2N6Na2O7S2 (Figura 1), o qual foi

Figura 1: Formula estrutural do corante remazol vermelho brilhante.

3.4. Saccharomyces cerevisiae

Saccharomycces cerevisiae é uma levedura, um fungo unicelular normalmente esférico ou

(a) (b) (c)

3.5. Coluna de Winogradsky

Inventada por Sergei Nikolaievich Winogradsky em 1880, a coluna forma um micro

Figura 4: Esquematização da coluna de Winogradsky.

3.6. Algas e fitoplâncton

As algas são organismos de natureza variada e ampla distribuição, estando presentes em

Devido a sua importância em relação a produção primária, o fitoplâncton é considerado

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