Microrganismos associados ao pólen fermentado de Melipona quadrifasciata (APIDAE, MELIPONINI)
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Data
2024-03-21
Autores
Câmera, Laís Calpacci [UNIFESP]
Orientadores
Morais, Michelle Manfrini [UNIFESP]
Tipo
Dissertação de mestrado
Título da Revista
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Resumo
As abelhas sem ferrão são importantes polinizadores e a relação entre elas e alguns microrganismos têm despertado interesse nos últimos anos. Em busca de recursos para a colônia, as abelhas coletam néctar e pólen, estocam em potes de cerume e com a ação de diversos microrganismos fermentam o alimento estocado. Por possuir funções distintas, a caracterização funcional de cada microrganismo participante na fermentação do pólen estocado permanece desconhecida. Pensando nisso, este trabalho teve como objetivo investigar os microrganismos encontrados no pólen estocado de abelhas Melipona quadrifasciata através de métodos dependentes de cultura e descrever a presença desses microrganismos no pólen, principalmente os potencialmente fermentadores. Além disso, comparar os microrganismos encontrados nos três estágios de fermentação do pólen (seco, pastoso ou líquido). Para isso, foram coletados potes de pólen fechados estocados de onze colônias de abelhas e semeados em condições de laboratório. Esses potes, apesar de fechados apresentavam o pólen em diferentes estágios de fermentação. Os microrganismos foram cultivados em seis diferentes meios de cultura, caracterizados macro e microscopicamente e separados de acordo com a característica do estágio de fermentação do pólen de origem. O DNA de cada microrganismo selecionado foi extraído, purificado e o sequenciamento foi feito no equipamento NextSeq (Illumina). Os oito microrganismos que foram cultivados provenientes do pólen coletado em estado seco foram exclusivos. Quatro morfotipos apareceram tanto na cultura proveniente do pólen em estado pastoso quanto no pólen líquido. Dos oito microrganismos que cresceram a partir do pólen pastoso, quatro foram exclusivos e dos 12 provenientes do pólen líquido, oito foram exclusivos. As bactérias Gram positivas do gênero Bacillos foram os microrganismos mais encontrados. Além desse gênero, também foram encontradas amostras do gênero Staphylococcus e foi possível identificar morfologicamente a levedura do gênero Rhodotorula. Este estudo contribui com a caracterização da microbiota das colônias de abelhas sem ferrão, revelando gêneros microbianos essenciais para sua saúde e manutenção. Tais achados podem contribuir não só com o desenvolvimento de rações fermentadas para essas abelhas, mas também para a criação de uma variedade de produtos e serviços inovadores na indústria biotecnológica, incluindo novos compostos antimicrobianos.
Stingless bees are essential pollinators, and the relationship between them and microorganisms has aroused interest in recent years. Looking for resources for the colony, the bees collect nectar and pollen, store them in cerumen pots, and ferment the stored food with the action of several microorganisms. As they have distinct functions, the functional characterization of each participating microorganism in the fermentation of stored pollen remains unknown. With this in mind, this work aims to investigate the microorganisms found in the stored pollen of Melipona quadrifasciata bees through culture-dependent methods and to describe their presence in pollen, especially those that are potentially fermenters. Furthermore, compare the microorganisms found in the three stages of pollen fermentation (dry, pasty or liquid). For this, closed pots of pollen stored from eleven bee colonies were seeded under laboratory conditions. These pots, despite being closed, showed the pollen at different stages of fermentation. The microorganisms were cultivated in six different culture media, characterized macro and microscopically and separated according to the characteristic of the fermentation stage of the pollen of origin. The DNA of each selected microorganism was extracted, purified, and the sequencing was made with the equipment NextSeq (Illumina). The eight microorganisms that were cultivated from pollen collected in a dry state did not appear in the other fermentation stages. Only four morphotypes appeared in both the culture from pasty pollen and liquid pollen. Of the eight microorganisms that grew from the pasty pollen, only four were exclusive and of the 12 from the liquid pollen, eight were exclusive. Most of the microorganisms found were Gram-positive bacteria of the genus Bacillus. Besides this genus, samples of the genus Staphylococcus were also found. It was also possible to morphologically identify the yeast of genus Rhodotorula. This work contributes to the microbiota characterization of the stingless bee colonies, showing microorganism genera that are essential for the health and maintenance of colonies. These discoveries have the potential to contribute not only to developing fermented feeds aimed at stingless bees, but also to creating a variety of innovative products and services in the biotechnology industry, including new antimicrobial compounds.
Stingless bees are essential pollinators, and the relationship between them and microorganisms has aroused interest in recent years. Looking for resources for the colony, the bees collect nectar and pollen, store them in cerumen pots, and ferment the stored food with the action of several microorganisms. As they have distinct functions, the functional characterization of each participating microorganism in the fermentation of stored pollen remains unknown. With this in mind, this work aims to investigate the microorganisms found in the stored pollen of Melipona quadrifasciata bees through culture-dependent methods and to describe their presence in pollen, especially those that are potentially fermenters. Furthermore, compare the microorganisms found in the three stages of pollen fermentation (dry, pasty or liquid). For this, closed pots of pollen stored from eleven bee colonies were seeded under laboratory conditions. These pots, despite being closed, showed the pollen at different stages of fermentation. The microorganisms were cultivated in six different culture media, characterized macro and microscopically and separated according to the characteristic of the fermentation stage of the pollen of origin. The DNA of each selected microorganism was extracted, purified, and the sequencing was made with the equipment NextSeq (Illumina). The eight microorganisms that were cultivated from pollen collected in a dry state did not appear in the other fermentation stages. Only four morphotypes appeared in both the culture from pasty pollen and liquid pollen. Of the eight microorganisms that grew from the pasty pollen, only four were exclusive and of the 12 from the liquid pollen, eight were exclusive. Most of the microorganisms found were Gram-positive bacteria of the genus Bacillus. Besides this genus, samples of the genus Staphylococcus were also found. It was also possible to morphologically identify the yeast of genus Rhodotorula. This work contributes to the microbiota characterization of the stingless bee colonies, showing microorganism genera that are essential for the health and maintenance of colonies. These discoveries have the potential to contribute not only to developing fermented feeds aimed at stingless bees, but also to creating a variety of innovative products and services in the biotechnology industry, including new antimicrobial compounds.
Descrição
Citação
Câmera, Laís Calpacci. Microrganismos associados ao pólen fermentado de Melipona quadrifasciata (APIDAE, MELIPONINI). Dissertação (Mestrado - Programa de Pós-Graduação em Mestrado em Biotecnologia) - Universidade Federal de São Paulo - Instituto de Ciência e Técnologia, São José dos Campos, 2024.