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Previous issue date: 2010-09-23 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Honey is a food produced by bees from the nectar of flowers collected and processed by the
digestive enzymes of insects, stored in combs in their hives to serve them food. The Brazilian
apiculture (branch of agriculture that studies of honey bees) has suffered high losses of honey
bees, due to a number of diseases that affect the hives, putting them at risk. Many species of
yeasts and molds can grow in the hive using honey as a substrate, such as the genus
Aspergillus, which is important because it contains species that produce mycotoxins and / or
pathogenic to bees. The Brazilian sac brood disease (BSB) is a disease with high mortality,
characterized by death in the pre-pupa or pupa and that have occurred in southeastern Brazil,
with substantial losses to beekeeping. However, its etiology is unknown in the region, which
were dismissed any similarity to the European Creates bagged caused by Sac Brood Virus
(SBV), and the Stryphnodendron polyphyllum (Fabaceae, Mimosoidea), the common name
barbatim?o. The objectives of this study were 1) to assess the fungal contamination in honey
bee brood and thus determining influence on the occurrence of BSB in the environment of the
hive, in Rio de Janeiro, 2) enumerate the fungal propagules in samples of honey in the comb,
and the young (pre-pupae and pupae) from apiaries located in regions affected by BSB, 3)
determine the frequency and identify the mycoflora total, 4) identify fungal species
pathogenic to bees, 5) characterize toxigenic profile isolated species of the genus Aspergillus.
A total of 43 honey samples and 43 samples of offspring (larvae and pupae) was purchased
from apiaries that have already expressed the CEB, in the municipalities of Barra do Pira?,
Mendes and Itaipava (endemic areas). Sampling occurred in the months corresponding to the
period before, during and after - disease (based on data from recent outbreaks). Analysis of
the mycoflora were performed by spread plate on the culture media dichloran rose bengal
chloramphenicol agar (DRBC) and dichloran 18% glycerol agar (DG18). The total fungal
counts were expressed as cfu g-1. We determined the number of samples and frequency of
isolation (%) of fungal genera and the relative density of species. The determination of
toxigenic fungi was done using the technique of thin layer chromatography (TLC). The values
of total fungal counts were similar in both media DRBC and DG18 for the samples of
offspring. The highest counts were observed in samples of honey in the comb, with 7,7 x 104
cfu g-1 on DRBC medium and 5.9 x 104 cfu g-1 in DG18 medium. Aspergillus, Penicillium
and Cladosporium were the most frequently isolated both in the honey comb, as in the
offspring. Aspergillus niger, A. flavus and Penicillium citrinum showed the highest relative
density in honey and brood. In the TLC analysis for the fungi isolated honey in the comb,
were not observed for positive strains isolated species of the genus Aspergillus. Among the
strains analyzed in the mycoflora of the larvae, there is presence of positive strains for
toxigenic profile of A. flavus in samples from Barra do Pirai and Itaipava. For strains
producing ochratoxins, the result was 100% negative for all sites studied. The fungi found in
this study can be related to losses bee in Rio de Janeiro, but is not present apparent direct with
BSB. However, more studies are needed to identify the mycoflora of honey and brood,
correlating the possible species that produce mycotoxins with the mycoflora present in the
substrate, promoting so help on the etiology of BSB in this state.
Keywords: fungi, honey, brood, bees. / O mel ? um alimento produzido pelas abelhas a partir do n?ctar recolhido de flores e
processado pelas enzimas digestivas desses insetos, sendo armazenado em favos em suas
colm?ias para servir-lhes de alimento. A apicultura brasileira tem sofrido altas perdas de
abelhas mel?feras, devido a uma s?rie de doen?as que afetam as colm?ias, colocando-as sob
risco. Muitas esp?cies de leveduras e fungos podem desenvolver-se na colm?ia utilizando o
mel como substrato, como por exemplo, o g?nero Aspergillus, importante por abranger
esp?cies produtoras de micotoxinas e/ou patog?nicas para as abelhas. A Cria Ensacada
Brasileira (CEB) ? uma doen?a com alto grau de mortalidade, caracterizada por morte na fase
de pr?-pupa ou pupa e que v?m ocorrendo na Regi?o sudeste do Brasil. Sua etiologia ?
desconhecida, j? que foi descartada qualquer semelhan?a com a Cria Ensacada Europ?ia
causada pelo Sac Brood V?rus (SBV), e com a intoxica??o pelo p?len do Stryphnodendron
polyphyllum, de nome vulgar barbatim?o. Os objetivos deste estudo foram: 1) avaliar a
contamina??o f?ngica em amostras de mel e crias de abelhas determinando assim rela??o com
a ocorr?ncia da CEB no ambiente da colm?ia, no Estado do Rio de Janeiro; 2) enumerar os
prop?gulos f?ngicos nas amostras de mel em favo, e de crias (pupas e pr?-pupas) provenientes
de api?rios localizados em regi?es acometidas pela CEB; 3) determinar a frequ?ncia e
identificar a micobiota total; 4) identificar esp?cies f?ngicas patog?nicas para as abelhas; 5)
caracterizar o perfil tox?geno de esp?cies isoladas do g?nero Aspergillus. Um total de 43
amostras de mel e 43 amostras de crias (larvas e pupas) foi adquirido em api?rios localizados
nos munic?pios de Barra do Pira?, Mendes e Itaipava (?reas end?micas). As coletas ocorreram
nos meses correspondentes ao per?odo pr?, e trans - doen?a (baseado nos dados dos ?ltimos
surtos). A an?lise da micobiota foi feita pelo m?todo de dilui??o em placa sobre os meios de
cultivo dicloran rosa bengala cloranfenicol agar (DRBC) e dicloran glicerol 18% agar
(DG18). As contagens f?ngicas totais foram expressas em ufc g-1. Foram determinadas o
n?mero de amostras e a freq??ncia de isolamento (%) dos g?neros f?ngicos e a densidade
relativa das esp?cies. A determina??o do perfil tox?geno dos fungos foi feita atrav?s da t?cnica
de cromatografia em camada delgada (CCD). Os valores de contagens f?ngicas totais foram
similares em ambos os meios DRBC e DG18, para as amostras de crias. As maiores
contagens foram observadas em amostras de mel em favo, com 7,7 x 104 ufc g-1 em meio
DRBC e 5,9 x 104 ufc g-1 em meio DG18. Aspergillus, Penicillium e Cladosporium foram os
g?neros mais freq?entemente isolados tanto no mel em favo, quanto nas crias. Aspergillus
niger, A. flavus e Penicillium citrinum apresentaram as maiores densidades relativas no mel e
nas crias. Na an?lise em CCD para os fungos isolados no mel em favo, n?o foram observadas
cepas positivas para as esp?cies isoladas do g?nero Aspergillus. Dentre as cepas analisadas na
micobiota das larvas, h? presen?a de cepas positivas para o perfil tox?geno de A. flavus nas
amostras de Barra do Pira? e Itaipava. Para as cepas produtoras de ocratoxinas, o resultado foi
de 100% negativas para todos os locais estudados. Os fungos encontrados neste estudo podem
estar relacionados ?s perdas ap?colas no estado do Rio de Janeiro, por?m n?o apresentam
aparente rela??o direta com a CEB. Contudo, mais estudos s?o necess?rios para identifica??o
da micobiota do mel e das crias, correlacionando ?s esp?cies poss?veis produtoras de
micotoxinas com a micobiota presente nos substratos, promovendo assim o aux?lio acerca da
etiologia da CEB neste estado.
| Identifer | oai:union.ndltd.org:IBICT/oai:localhost:tede/734 |
| Date | 23 September 2010 |
| Creators | Marassi, Ana Cl?udia |
| Contributors | Rosa, Carlos Alberto Rocha |
| Publisher | Universidade Federal Rural do Rio de Janeiro, Curso de P?s-Gradua??o em Ci?ncias Veterin?rias, UFRRJ, Brasil, Parasitologia Veterin?ria |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFRRJ, instname:Universidade Federal Rural do Rio de Janeiro, instacron:UFRRJ |
| Rights | info:eu-repo/semantics/openAccess |
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