Assessing Sperm Quality in Stingless Bees

Although stingless bees have a great potential as commercial pollinators, their exploitation depends on the successful reproduction of colonies on a large scale. To do so, it is essential to develop accurate diagnostic tools that facilitate a better understanding of the reproductive biology of stingless bees. Sperm counts, sperm morphology and sperm viability (the relative proportion of live to dead sperm), are key parameters assessing semen quality and potential fertilization success. Here we present standardized protocols to assess these three parameters. We used Scaptotrigona aff. depilis (Moure) as a study model. Semen extractions from the seminal vesicles were found to yield better results when performed in mature rather than in younger males. For morphology and viability analyses, the best semen dilution on Hayes solution was adding 120 µl to the contents of the two seminal vesicles. For sperm count, however, we recommend a higher dilution (1.000 µl). Sperm viability values were higher when Hayes solution was adjusted to pH 8.7 and no difference were found in sperm viability when analyzing fresh semen, and semen collected 1, 3 and 5 hours prior to the analyzes. However, we found a significant decrease in sperm viability when analyzing sperm collected 24 hours beforehand. The protocols seem sensitive and efficient, and show great potential to be used in other stingless bee species

¿Cómo convertirse en un ciudadano científico?

La Ciencia Ciudadana nos brinda una oportunidad de conocer más a la naturaleza, aprender sobre ella, contribuir a la ciencia, compartir nuestras observaciones y motivar a otras personas a interesarse por la biodiversidad que nos rodea. Además, la ciencia ciudadana puede contribuir a la elaboración de políticas públicas y leyes, y fomentar la participación de la sociedad en la toma de decisiones. ¡Hay muchas razones para participar en proyectos de Ciencia Ciudadana!Fil: Arbetman, Marina Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; ArgentinaFil: Barahona Segovia, Rodrigo M.. Universidad de Los Lagos; ChileFil: Burgos, Adriana. Jardin Botánico Carlos Thays; ArgentinaFil: Aparecida Gobatto, Alexandra. Instituto de Pesquisas Jardim Botânico do Rio de Janeiro; BrasilFil: Koffler, Sheina. Universidade de Sao Paulo; BrasilFil: Queiroz Souza, Caren. Universidade Federal da Bahia; Brasi

Flower-Insect Timed Counts (FIT Count):protocol adaptation and preliminary results in Brazil

FIT Count (Flower-Insect Timed Counts) is a citizen science initiative that seeks to monitor flower visitation by diverse pollinator groups, including bumblebees, stingless bees, honeybees, flies, hummingbirds, among others. The protocol entails determining a 50 x 50 cm plot area around a target plant species, photographing the plant and conducting a standardized 10-minute survey on this area to estimate the frequency of flower visits by different pollinator groups. Conducting FIT Counts in different habitats and locations produces data on the temporal and spatial dynamics of these interactions (Carvell 2022, UK Pollinator Monitoring Scheme 2018).FIT Count methodology was originally developed by the UK Pollinator Monitoring Scheme (PoMS) and has been used in the United Kingdom since 2017, with over 8,500 counts conducted to date. In 2021, the FIT Count application (app) was launched in the United Kingdom, and became available in Brazil, Chile, Argentina, and some other European countries in 2022. After installation, users may select in which country the observations will be conducted and their preferred language, either English or the main native language of participating countries. The adaptation of the application for use by Brazilian citizen scientists involved not only the translation of the interface, but also relied on the expertise of a local team who helped select which plant species and pollinator groups would be appropriate and representative of biodiversity within the country (Fig. 1). The application is supported by a website that features a dedicated page for each country. Users can download the app on Google Play or App Store (Fig. 2).To allow the participation of non-experts, who generally are not familiar with taxonomy or species identification, the flower visitors are identified in general categories such as bumblebees and carpenter bees (grouped together), flies, and others (see Fig. 1), representing the main functional groups of pollinators. If a flower visitor cannot be identified, it should be classified as "other insect" or marked as "I don't know". Including the possibility of uncertainty in flower visitor identification was a suggestion of the Brazilian team for quality assurance during data collection.Currently, 834 flower visits have been counted by 15 participants, who performed 109 FIT Counts across Brazil as a pilot testing phase of the app. Two training workshops were held to disseminate the app, and an illustrated guide was published to help participants identify flower types and flower visitor groups (Koffler 2022). Nineteen plant species were monitored, but most flower-visits counted were on basil (Ocimum basillicum, n = 47) and false heather (Cuphea hyssopifolia, n = 33). The mean number of flower visitors per FIT Count was 8 (min = 0, max = 45). While most visits were performed by honeybees, participants also recorded visits by beetles, butterflies or moths, small insects, solitary bees, wasps and other insects. Next steps include assessing data quality and promoting strategic partnerships to further disseminate this citizen science initiative. For instance, quality control measures may include assessing whether participants perform the protocol steps according to instructions and whether plants and flower visitors are correctly identified and counted. Also, we are exploring the prospective implementation of the Darwin Core standard (Darwin Core Task Group 2009) along with the Plant-Pollinator Interactions vocabulary (Salim 2022) to standardize data description

Opportunities and Barriers for Citizen Science Growth in Brazil: Reflections from the First Workshop of the Brazilian Citizen Science Network

Citizen science networks can provide support and visibility to existing initiatives. Although citizen science is a growing practice in Brazil, several challenges still exist. A network can play an important role in overcoming these challenges, helping with the exchange of knowledge between initiatives or raising funding, for example. The Brazilian Citizen Science Network was created to advance citizen science in the country. The launch took place in March 2021 at a two-day online workshop focused on topics such as financial sustainability, technology, data quality, inter- and transdisciplinary infrastructure, engagement, and good practices. Based on the contents of the presentations and comments during the event, the authors of this report held follow-up meetings to identify favourable and unfavourable factors that can foster or constrain citizen science in Brazil. Afterwards, an open letter expressing these factors was produced and shared with the network members to register the status of challenges and opportunities we have discussed so far, to be used as a guide for future actions. This report provides an overview of the workshop. Despite depicting a local perspective, the challenges and opportunities discussed during the workshop and summarised in this report may also apply to fostering citizen science in other countries or establishing global practices. We consider that the Brazilian Citizen Science Network can play an important role in promoting the establishment of collaborative actions across the country

Flower-insect timed counts (FIT Count): protocol adaptation and preliminary results in Brazil

FIT Count (Flower-Insect Timed Counts) is a citizen science initiative that seeks to monitor flower visitation by diverse pollinator groups, including bumblebees, stingless bees, honeybees, flies, hummingbirds, among others. The protocol entails determining a 50 x 50 cm plot area around a target plant species, photographing the plant and conducting a standardized 10-minute survey on this area to estimate the frequency of flower visits by different pollinator groups. Conducting FIT Counts in different habitats and locations produces data on the temporal and spatial dynamics of these interactions

Citizen Science Data on Urban Forageable Plants:A Case Study in Brazil

This paper presents two key data sets derived from the Pomar Urbano project. The first data set is a comprehensive catalog of edible fruit-bearing plant species, native or introduced in Brazil. The second data set, sourced from the iNaturalist platform, tracks the distribution and monitoring of these plants within urban landscapes across Brazil. The study encompasses data from all 27 Brazilian state capitals, focusing on the ten cities that contributed the most observations as of August 2023. The research emphasizes the significance of citizen science in urban biodiversity monitoring and its potential to contribute to various fields, including food and nutrition, creative industry, study of plant phenology, and machine learning applications. We expect the data sets to serve as a resource for further studies in urban foraging, food security, cultural ecosystem services, and environmental sustainability

Optimizing the Monitoring of Urban Fruit-Bearing Flora with Citizen Science:An Overview of the Pomar Urbano Initiative

The "Pomar Urbano" (Urban Orchard) project focuses on the collaborative monitoring of fruit-bearing plant species in urban areas throughout Brazil.The project collected a list of 411 fruit-bearing plant species (Soares et al. 2023), both native and exotic varieties found in Brazil. This list was selected from two main sources: the book Brazilian Fruits and Cultivated Exotics (Lorenzi et al. 2006) and the book series Plants for the Future, which includes volumes specifically dedicated to species of economic value in different regions of Brazil, namely the South (Coradin et al. 2011), Midwest (Vieira et al. 2016), Northeast (Coradin et al. 2018) and North (Coradin et al. 2022). To ensure broad geographic coverage, the project spans all 27 state capitals of Brazil. The data collection process relies on the iNaturalist Umbrella and Collection projects. Each state capital has a single collection project, including the fruit-bearing plant species list, and the locality restriction to that specific city. For example, the collection project Pomar Paulistano gathers data from the city of São Paulo. The Umbrella Project Urban Orchard was set to track data from all 27 collection projects.We firmly believe that these fruit-bearing plant species possess multifaceted value that extends beyond mere consumption. As such, we have assembled a dynamic and multidisciplinary team comprising professionals from various institutions across Brazil in a collaborative effort that encompasses different dimensions of biodiversity value exploration and monitoring, especially phenological data.One facet of our team is focused on creating products inspired by the diverse array of Brazilian fruit-bearing plants. Their work spans across sectors of the creative industry, including fashion, painting, and graphic design to infuse these natural elements into innovative and sustainable designs (Fig. 1 and Fig. 2).A group of nutrition and health scientists in conjunction with communication and marketing professionals is working to produce engaging media content centered around food recipes that incorporate Brazilian fruits (Fig. 3). These recipes primarily feature the fruit-bearing plants most frequently observed on iNaturalist in the city of São Paulo, allowing us to showcase the local biodiversity while promoting culinary diversity. Some of these recipes are based on the book Brazilian Biodiversity: Flavors and Aromas (Santiago and Coradin 2018). This book is an extensive compendium of food recipes that use fruits derived from native Brazilian species

Leveraging citizen science for monitoring urban forageable plants

Urbanization brings forth social challenges in emerging countries such as Brazil, encompassing food scarcity, health deterioration, air pollution, and biodiversity loss. Despite this, urban areas like the city of São Paulo still boast ample green spaces, offering opportunities for nature appreciation and conservation, enhancing city resilience and livability. Citizen science is a collaborative endeavor between professional scientists and nonprofessional scientists in scientific research that may help to understand the dynamics of urban ecosystems. We believe citizen science has the potential to promote human and nature connection in urban areas and provide useful data on urban biodiversity

Influence of the amount of food resources and seasonality on the production of sexual offspring in Scaptotrigona aff. depilis (Apidae, Meliponini)

Em colônias de insetos eussociais, é prevista uma demanda conflitante entre a produção de operárias e indivíduos reprodutivos, visto que operárias realizam tarefas essenciais para a sobrevivência da colônia, enquanto machos e rainhas estão relacionados com a reprodução. Nesse trabalho, foram testadas as hipóteses (1) que a produção de sexuados estaria diretamente relacionada com a disponibilidade de recursos alimentares e (2) que haveria uma regulação temporal da produção de sexuados, com épocas mais favoráveis à produção de reprodutivos (verão) e menos favoráveis (inverno). O sistema de estudo foi a abelha sem ferrão Scaptotrigona aff. depilis e foram realizados dois experimentos. No primeiro experimento, na estação de verão, foram montadas quinze colônias padronizadas e sorteadas em cinco grupos experimentais. Os grupos A2 e C2 receberam o dobro de alimentação artificial (xarope de açúcar e pólen apícola) do que os grupos A1 e C1. As colônias dos grupos C2 e C1 foram mantidas confinadas, enquanto as colônias dos grupos A2 e A1 podiam forragear livremente. O grupo A0 (controle) não recebeu alimentação artificial e as colônias permaneceram abertas. No inverno, o mesmo experimento foi realizado, porém apenas com os grupos com colônias abertas. No experimento 2, a quantidade de pólen estocado nas colônias foi manipulada. As colônias do grupo Co não foram manipuladas e apresentaram em média 44 potes de pólen, as colônias do grupo A receberam em média 96 potes e as colônias do grupo B, 12 potes. No experimento 1, para o verão, foi encontrado um efeito significativo do confinamento, porém, nas duas estações, não houve efeito da quantidade de alimento sobre a produção de sexuados. Nas duas estações, verificou-se grande variação dentro dos grupos e a produção de machos foi dominada por poucas colônias. Não houve diferenças em relação à produção de sexuados entre as estações. No inverno, a produção de machos e células reais foi correlacionada positivamente com a quantidade de potes de alimento nas colônias. No experimento 2, apesar das diferenças iniciais quanto ao alimento armazenado, houve uma rápida recuperação dos estoques de alimento, principalmente pelo grupo B. Não foi verificada diferença na produção de sexuados devido ao tratamento experimental, porém, a produção de machos foi correlacionada com o número de potes de pólen nos ninhos, como no experimento 1. Considerando esses resultados em conjunto, não foi possível refutar as hipóteses de efeito positivo da quantidade de recursos alimentares e de efeito da sazonalidade sobre a produção de sexuados em S. aff. depilis.In colonies of eusocial insects, it is expected a trade-off in resource allocation between worker and sexual offspring production, as workers perform tasks essential to colony\'s survival, while males and queens are associated with reproduction. In this study, we tested the following hypotheses: (1) that sexual offspring production would be directly related to food resources availability and (2) that there would be a temporal regulation of sexual offspring production, with a favorable season to reproductives production (summer) and a less favorable season (winter). The study system was the stingless bee Scaptotrigona aff. depilis and two experiments were conducted. In the first experiment, in summer, fifteen standardized colonies were set up and randomly assigned into five groups. Groups A2 and C2 received twice the amount of artificial food (sugar syrup and Apis mellifera pollen) than colonies from groups A1 and C1. Colonies from groups C2 and C1 were kept confined, while colonies from groups A2 and A1 could forage freely. The control group A0 did not receive any artificial food and the colonies remained open. In the winter the same experiment was carried out, only with groups with open colonies. In experiment 2, the amount of pollen stored in the colonies was manipulated. The colonies of group Co were not manipulated and presented a mean number of 44 pots of pollen, colonies from group A received 96 pots and colonies from group B 12 pots (mean). In summer (experiment 1), there was a significant effect for confinement, however for both seasons, there was no effect for food amount on sexual offspring production. For both seasons, there was a large intra-group variation and a few colonies were responsible for major male production. There was no difference on sexual offspring production between seasons. In winter, the production of males and royal cells was positively correlated with the amount of food pots in the colonies. In experiment 2, even though there was an initial difference in relation to food stores, a fast recovery on food storage was observed, specially for group B. There was no difference on sexual offspring production due to experimental treatment, however male production was correlated with the number of pollen pots, as in experiment 1. Considering these results together, it was not possible to refute the hypotheses of the positive effect of amount of food resources and the effect of seasonality on the production of sexual offspring in S. aff. depilis

Avaliação da biologia reprodutiva, genética quantitativa e consequências do manejo a longo prazo em abelhas sem ferrão para subsidiar programas de melhoramento genético

Meliponiculture (or stingless beekeeping) is a powerful tool for sustainable economic development in the tropics. However, meliponiculture has many challenges as it lacks standardization in management and breeding practices. The aim of this thesis was to investigate stingless bee reproductive biology combined to management and breeding as background to meliponiculture improvement. In chapter 1, we performed a meta-analysis of heritability across the Hymenoptera in order to review current knowledge and discuss the challenges for bee breeding and conservation. In chapter 2, we assessed how sexual selection acts on male traits in the stingless bee Scaptotrigona aff. depilis and identified which traits may affect male fitness. In chapter 3, we estimated heritability and genetic correlations for the traits studied in the previous chapter, to understand how evolution shapes male traits and to identify important traits for breeding programs. Finally, in chapter 4 we investigated how the environment and long-term management influence colony productivity in Melipona subnitida (jandaíra), a commercially managed species in Northeastern Brazil. Our results revealed that morphological traits exhibit higher heritability estimates than fitness related traits, and in general, colony productivity traits showed potential for breeding. However, few studies are available for stingless bees yet. Male aggregations in S. aff depilis selected competitive males with higher quality sperm, indicating the importance of this mechanism in stingless bee mating biology. The studied male traits exhibited high heritability estimates, with exception of sperm length. Since aggregations selected males with shorter sperm, these results suggest selection for long-term sperm storage by queens and higher fertilization potential. The assessment of M. subnitida records revealed that honey production was affected by climate and management, and strategies to improve beekeeping practices were discussed. We believe this thesis provides important guidelines to establish successful stingless bee breeding programsA meliponicultura, criação racional de abelhas sem ferrão, é uma poderosa ferramenta para o desenvolvimento econômico sustentável em regiões tropicais. No entanto, a prática da meliponicultura enfrenta diversos desafios, como a falta de padronização das técnicas de manejo e melhoramento genético. O objetivo desta tese foi investigar a biologia reprodutiva de abelhas sem ferrão aliada ao manejo e ao melhoramento genético, a fim de fornecer subsídios para o aprimoramento da meliponicultura. No capítulo 1, realizamos uma meta-análise sobre herdabilidade em Hymenoptera, a fim de compreender o conhecimento atual e os desafios associados ao melhoramento genético de abelhas e sua conservação. No capítulo 2, avaliamos como a seleção sexual atua em caracteres dos machos da espécie Scaptotrigona aff. depilis e identificamos quais caracteres podem influenciar o sucesso reprodutivo. No capítulo 3, estimamos a herdabilidade e correlações genéticas para os caracteres avaliados no capítulo anterior, a fim de entender como a evolução atua moldando os caracteres dos machos e quais desses caracteres podem ser utilizados em programas de melhoramento genético. Finalmente, no capítulo 4, investigamos o efeito do ambiente e do manejo na produtividade de colônias em Melipona subnitida (jandaíra), uma espécie manejada comercialmente no Nordeste brasileiro. Nossos resultados revelaram que caracteres morfológicos exibem estimativas de herdabilidade mais altas do que caracteres ligados ao sucesso reprodutivo. No entanto, poucos estudos com abelhas sem ferrão foram realizados até o momento. Agregações de machos em S. aff. depilis selecionaram machos mais competitivos que apresentaram maior qualidade espermática, indicando a importância desse mecanismo na biologia reprodutiva de abelhas sem ferrão. Os caracteres estudados apresentaram alta herdabilidade, com exceção do comprimento do espermatozoide. Como agregações selecionam machos com espermatozoides mais curtos, esses resultados sugerem seleção direcionada para um maior tempo de armazenamento do esperma pelas rainhas e maior potencial de fertilização. A avaliação dos registros de M. subnitida revelou que a produção de mel foi afetada pelo clima e pelo manejo, e estratégias a fim de melhorar as práticas da meliponicultura foram discutidas. Acreditamos que essa tese fornece importantes resultados para o estabelecimento de programas de melhoramento genético em abelhas sem ferrã