Convolutional Neural Networks for Real Time Classification of Beehive Acoustic Patterns on Constrained Devices.

Recent research has demonstrated the effectiveness of convolutional neural networks (CNN) in assessing the health status of bee colonies by classifying acoustic patterns. However, developing a monitoring system using CNNs compared to conventional machine learning models can result in higher computation costs, greater energy demand, and longer inference times. This study examines the potential of CNN architectures in developing a monitoring system based on constrained hardware. The experimentation involved testing ten CNN architectures from the PyTorch and Torchvision libraries on single-board computers: an Nvidia Jetson Nano (NJN), a Raspberry Pi 5 (RPi5), and an Orange Pi 5 (OPi5). The CNN architectures were trained using four datasets containing spectrograms of acoustic samples of different durations (30, 10, 5, or 1 s) to analyze their impact on performance. The hyperparameter search was conducted using the Optuna framework, and the CNN models were validated using k-fold cross-validation. The inference time and power consumption were measured to compare the performance of the CNN models and the SBCs. The aim is to provide a basis for developing a monitoring system for precision applications in apiculture based on constrained devices and CNNs.

Discrete Time Series Forecasting of Hive Weight, In-Hive Temperature, and Hive Entrance Traffic in Non-Invasive Monitoring of Managed Honey Bee Colonies: Part I.

From June to October, 2022, we recorded the weight, the internal temperature, and the hive entrance video traffic of ten managed honey bee (Apis mellifera) colonies at a research apiary of the Carl Hayden Bee Research Center in Tucson, AZ, USA. The weight and temperature were recorded every five minutes around the clock. The 30 s videos were recorded every five minutes daily from 7:00 to 20:55. We curated the collected data into a dataset of 758,703 records (280,760-weight; 322,570-temperature; 155,373-video). A principal objective of Part I of our investigation was to use the curated dataset to investigate the discrete univariate time series forecasting of hive weight, in-hive temperature, and hive entrance traffic with shallow artificial, convolutional, and long short-term memory networks and to compare their predictive performance with traditional autoregressive integrated moving average models. We trained and tested all models with a 70/30 train/test split. We varied the intake and the predicted horizon of each model from 6 to 24 hourly means. Each artificial, convolutional, and long short-term memory network was trained for 500 epochs. We evaluated 24,840 trained models on the test data with the mean squared error. The autoregressive integrated moving average models performed on par with their machine learning counterparts, and all model types were able to predict falling, rising, and unchanging trends over all predicted horizons. We made the curated dataset public for replication.

Post-transcriptional regulation of Dufour's gland reproductive signals in bumble bees.

Pheromone communication is a key mechanism by which the reproductive division of labor is maintained within insect communities. Understanding how pheromones evolved to regulate social behavior requires knowledge of the molecular regulation of their production. However, even in cases where pheromones were identified, our understanding of their biosynthesis and molecular regulation remains limited. Bumble bees provide a unique system to explore pheromone biosynthesis since workers produce ester sterility signals in their Dufour's gland that differ from gyne-specific esters and are not produced by queens. These esters are hypothesized to be produced in the exocrine gland where they are stored, and indeed queens, gynes and workers differ significantly in the expression of Dufour's gland genes coding to enzymes involved in the biosynthesis of esters. However, a previous transcriptome analysis revealed no gene expression differences in the Dufour's gland of workers despite differences in both ester production and ovarian activation, suggesting that ester production may be regulated lower down. Proteomics of the Dufour's gland of queens, gynes, and workers recovered over 2400 proteins and broadly matched the previous RNAseq data. However, more than 100 differentially expressed proteins were found between the worker groups, including key enzymes in fatty acid biosynthesis, indicating that the regulation of reproductive signal biosynthesis in workers is done post-transcription. Overall, our data provide evidence that pheromone biosynthesis in the Dufour's gland is caste specific, that gynes and workers are likely using different enzymes to make their respective wax esters, and that the regulation on pheromone production in queens, gynes and workers is likely done at different regulatory levels, with workers signals being subjected to regulation at the protein level.

Foraging distance distributions reveal how honeybee waggle dance recruitment varies with landscape.

Honeybee (Apis mellifera) colonies use a unique collective foraging system, the waggle dance, to communicate and process the location of resources. Here, we present a means to quantify the effect of recruitment on colony forager allocation across the landscape by simply observing the waggle dance on the dancefloor. We show first, through a theoretical model, that recruitment leaves a characteristic imprint on the distance distribution of foraging sites that a colony visits, which varies according to the proportion of trips driven by individual search. Next, we fit this model to the real-world empirical distance distribution of forage sites visited by 20 honeybee colonies in urban and rural landscapes across South East England, obtained via dance decoding. We show that there is considerable variation in the use of dancing information in colony foraging, particularly in agri-rural landscapes. In our dataset, reliance on dancing increases as arable land gives way to built-up areas, suggesting that dancing may have the greatest impact on colony foraging in the complex and heterogeneous landscapes of forage-rich urban areas. Our model provides a tool to assess the relevance of this extraordinary behaviour across modern anthropogenic landscape types.

Bioactive compounds enrichment in rabbit doe's diet pre-and during pregnancy improves productive and reproductive performance and cost-effectiveness under hot climates.

This study aimed to investigate the effect of diet supplementation with a symbiotic (SY), bee pollen (BP), honey bee (HB), date palm pollen (DPP) and their mixture (MIX) on female rabbit productive and reproductive performances under desert hot climates. Seventy-two Californian does of 5 months age and average body weight of 3250 ± 78.2 g were randomly allotted into six groups, each of 12 does. All does orally receive 3 ml distilled water for 10 days before mating and 28 days during pregnancy. Treatments were repeated for four consecutive parities. The first group served as control (C) given distilled water only, however the second, third, fourth and fifth groups were supplemented with 3 ml distilled water containing 0.2 ml SY, 200 mg DPP, 200 mg BP, 0.2 ml HB/doe per day, respectively. While, the sixth group does were given all previous ingredients (MIX). Sexual receptivity rate, fertility rate, kindling rate, and newborn traits were recorded. Also, maternal feed intake, feed conversion ratio, and digestibility coefficients of nutrients were recorded. Does in all groups were artificially inseminated with 0.5 ml of fresh heterospermic semen of 15 fertile bucks extended in Tris at 806-1006 sperm/ml. Treatment increased maternal body weight and daily gain with highest values (P < 0.05) in BP, SY, and MIX does. All treatments enhanced feed intake and feed conversion ratio (FCR) compared with control. Number services per conception decreased (P < 0.01), while litter size and weight and survival at birth and weaning increased (P < 0.01) in treated than control does. Treated does produced more milk than control. Digestibility coefficients of all nutrients were improved (P < 0.01) in treated does. In conclusion, supporting rabbit does pre- and during pregnancy with diets supplemented with a mixture of honey bee, date palm pollen, bee pollen, and synbiotic improves the productive and reproductive performances of rabbit does and their offspring.

Neither nectar nor deception: the role of floral rewards in the pollination system of Laelia (Orchidaceae: Laeliinae).

Orchids offer a variety of floral rewards to pollinators. In many orchid groups, however, the transfer of pollen is based on food-deception, as in the case of Laelia (including Schomburgkia s.s.), a genus assigned to the Neotropical subtribe Laeliinae. Here, we report on the reproductive biology of a Brazilian member of this subtribe, namely, Laelia gloriosa, occurring in the forested areas of southeastern Brazil. The study includes analyses of floral morphology, histochemistry, and the chemical analysis of floral rewards and scents. Pollinators and pollination mechanism data were collected in the field by means of focal observations. Analyses of breeding systems and the percentage of potentially viable seed were also recorded. The floral morphology of Laelia gloriosa indicates that this species is melittophilous. The flowers release a citrus-like fragrance that attracts many species of bee. The flowers offer waxy material as a reward, and this is collected exclusively by Meliponini bees. Several bee species visit the flowers. However, those of L. gloriosa are pollinated exclusively by Trigona spinipes. Pollinaria are deposited on the bee's scutellum. Plants of the studied population were entirely self-compatible, but pollinator-dependent. The frequency of visits to these flowers was greater than in rewardless Laeliinae. Our study provides the first evidence of lipoidal substances as a resource in Laeliinae. The discovery that an orchid species (Laelia gloriosa) of this subtribe, hitherto considered to be entirely pollinated by nectar-seeking pollinators, offers a floral waxy material and provides new insights into the evolution of this important subtribe.

Physically stressed bees expect less reward in an active choice judgement bias test.

Emotion-like states in animals are commonly assessed using judgment bias tests that measure judgements of ambiguous cues. Some studies have used these tests to argue for emotion-like states in insects. However, most of these results could have other explanations, including changes in motivation and attention. To control for these explanations, we developed a novel judgment bias test, requiring bumblebees to make an active choice indicating their interpretation of ambiguous stimuli. Bumblebees were trained to associate high or low rewards, in two different reward chambers, with distinct colours. We subsequently presented bees with ambiguous colours between the two learnt colours. In response, physically stressed bees were less likely than control bees to enter the reward chamber associated with high reward. Signal detection and drift diffusion models showed that stressed bees were more likely to choose low reward locations in response to ambiguous cues. The signal detection model further showed that the behaviour of stressed bees was explained by a reduction in the estimated probability of high rewards. We thus provide strong evidence for judgement biases in bees and suggest that their stress-induced behaviour is explained by reduced expectation of higher rewards, as expected for a pessimistic judgement bias.

Ecological principles for the evolution of communication in collective systems.

Communication allows members of a collective to share information about their environment. Advanced collective systems, such as multicellular organisms and social insect colonies, vary in whether they use communication at all and, if they do, in what types of signals they use, but the origins of these differences are poorly understood. Here, we develop a theoretical framework to investigate the evolution and diversity of communication strategies under collective-level selection. We find that whether communication can evolve depends on a collective's external environment: communication only evolves in sufficiently stable environments, where the costs of sensing are high enough to disfavour independent sensing but not so high that the optimal strategy is to ignore the environment altogether. Moreover, we find that the evolution of diverse signalling strategies-including those relying on prolonged signalling (e.g. honeybee waggle dance), persistence of signals in the environment (e.g. ant trail pheromones) and brief but frequent communicative interactions (e.g. ant antennal contacts)-can be explained theoretically in terms of the interplay between the demands of the environment and internal constraints on the signal. Altogether, we provide a general framework for comparing communication strategies found in nature and uncover simple ecological principles that may contribute to their diversity.

Variation in season length and development time is sufficient to drive the emergence and coexistence of social and solitary behavioural strategies.

Season length and its associated variables can influence the expression of social behaviours, including the occurrence of eusociality in insects. Eusociality can vary widely across environmental gradients, both within and between different species. Numerous theoretical models have been developed to examine the life history traits that underlie the emergence and maintenance of eusociality, yet the impact of seasonality on this process is largely uncharacterized. Here, we present a theoretical model that incorporates season length and offspring development time into a single, individual-focused model to examine how these factors can shape the costs and benefits of social living. We find that longer season lengths and faster brood development times are sufficient to favour the emergence and maintenance of a social strategy, while shorter seasons favour a solitary one. We also identify a range of season lengths where social and solitary strategies can coexist. Moreover, our theoretical predictions are well matched to the natural history and behaviour of two flexibly eusocial bee species, suggesting that our model can make realistic predictions about the evolution of different social strategies. Broadly, this work reveals the crucial role that environmental conditions can have in shaping social behaviour and its evolution and it underscores the need for further models that explicitly incorporate such variation to study the evolutionary trajectories of eusociality.

Novel fungicide and neonicotinoid insecticide impair flight behavior in pollen foraging honey bees, Apis mellifera.

Bees are often exposed to pesticides affecting physiological functions and molecular mechanisms. Studies showed a potential link between altered expression of energy metabolism related transcripts and increased homing flight time of foragers exposed to pesticides. In this study, we investigated the effects of thiamethoxam and pyraclostrobin on longevity, flight behavior, and expression of transcripts involved in endocrine regulation (hbg-3, buffy, vitellogenin) and energy metabolism (cox5a, cox5b, cox17) using radio frequency identification (RFID) technology and quantitative polymerase chain reaction. Parallel, a laboratory study was conducted investigating whether pesticide exposure alone without the influence of flight activity caused similar expression patterns as in the RFID experiment. No significant effect on survival, homing flight duration, or return rate of exposed bees was detected. The overall time foragers spent outside the hive was significantly reduced post-exposure. Irrespective of the treatment group, a correlation was observed between cox5a, cox5b, cox17 and hbg-3 expression and prolonged homing flight duration. Our results suggest that flight behavior can impact gene expression and exposure to pesticides adversely affects the expression of genes that are important for maintaining optimal flight capacity. Our laboratory-based experiment showed significantly altered expression levels of cox5a, cox6c, and cox17. However, further work is needed to identify transcriptional profiles responsible for prolonged homing flight duration.

High-resolution outdoor videography of insects using Fast Lock-On tracking.

Insects have important roles globally in ecology, economy, and health, yet our understanding of their behavior remains limited. Bees, for example, use vision and a tiny brain to find flowers and return home, but understanding how they perform these impressive tasks has been hampered by limitations in recording technology. Here, we present Fast Lock-On (FLO) tracking. This method moves an image sensor to remain focused on a retroreflective marker affixed to an insect. Using paraxial infrared illumination, simple image processing can localize the sensor location of the insect in a few milliseconds. When coupled with a feedback system to steer a high-magnification optical system to remain focused on the insect, a high-spatiotemporal resolution trajectory can be gathered over a large region. As the basis for several robotic systems, we show that FLO is a versatile idea that can be used in combination with other components. We demonstrate that the optical path can be split and used for recording high-speed video. Furthermore, by flying an FLO system on a quadcopter drone, we track a flying honey bee and anticipate tracking insects in the wild over kilometer scales. Such systems have the capability to provide higher-resolution information about insects behaving in natural environments and as such will be helpful in revealing the biomechanical and neuroethological mechanisms used by insects in natural settings.

Autonomous tracking of honey bee behaviors over long-term periods with cooperating robots.

Digital and mechatronic methods, paired with artificial intelligence and machine learning, are transformative technologies in behavioral science and biology. The central element of the most important pollinator species-honey bees-is the colony's queen. Because honey bee self-regulation is complex and studying queens in their natural colony context is difficult, the behavioral strategies of these organisms have not been widely studied. We created an autonomous robotic observation and behavioral analysis system aimed at continuous observation of the queen and her interactions with worker bees and comb cells, generating behavioral datasets of exceptional length and quality. Key behavioral metrics of the queen and her social embedding within the colony were gathered using our robotic system. Data were collected continuously for 24 hours a day over a period of 30 days, demonstrating our system's capability to extract key behavioral metrics at microscopic, mesoscopic, and macroscopic system levels. Additionally, interactions among the queen, worker bees, and brood were observed and quantified. Long-term continuous observations performed by the robot yielded large amounts of high-definition video data that are beyond the observation capabilities of humans or stationary cameras. Our robotic system can enable a deeper understanding of the innermost mechanisms of honey bees' swarm-intelligent self-regulation. Moreover, it offers the possibility to study other social insect colonies, biocoenoses, and ecosystems in an automated manner. Social insects are keystone species in all terrestrial ecosystems; thus, developing a better understanding of their behaviors will be invaluable for the protection and even the restoration of our fragile ecosystems globally.

Biodiversity measures of a grassland plant-pollinator community are resilient to the introduction of honey bees (Apis mellifera).

The prairies of Canada support a diversity of insect pollinators that contribute pollination services to flowering crops and wild plants. Habitat loss and use of managed pollinators has increased conservation concerns for wild pollinators, as mounting evidence suggests that honey bees (Apis mellifera) may reduce their diversity and abundance. Plant-pollinator community analyses often omit non-bee pollinators, which can be valuable contributors to pollination services. Here, we experimentally introduced honey bees to examine how their abundance affects the species richness, diversity, abundance, species composition, interaction richness, and interaction diversity of all wild pollinators, and of four higher taxa separately. We identified all insect pollinators and analyzed how honey bee abundance affected the above biodiversity metrics, controlling for flower abundance and flower species richness. Even with high honey bee densities, there was no change to any of these variables, except that beetle species diversity increased. All other taxa had no significant relationship to honey bee abundance. Considering the widespread use of managed honey bees, the effect they have on wild pollinators should be firmly established. Our results suggest that honey bees have little to no short-term impact on the wild pollinator community or its interactions with plants in this native grassland.

Unique brood ester profile in a Varroa destructor resistant population of European honey bee (Apis mellifera).

Varroa destructor is one of the greatest threats to Apis mellifera worldwide and if left untreated will kill a colony in less than three years. A Varroa-resistant population from Gotland, Sweden, has managed to survive for 25 years with little to no Varroa treatment by reducing the mite's reproductive success. The underlying mechanisms of this trait is currently not known, though previous research indicates that it is the honey bee brood, and not adult bee influence, that contributes to this phenotype. As the mite's own reproduction is synchronized with the brood's development though the interception of brood pheromones, it is possible that a change in pheromone profile would disrupt the mite's reproductive timing. To investigate this, we characterized the brood ester pheromone (BEP) profile of our resistant Gotland population compared to a non-resistant control. This was done by extracting and analyzing key cuticular compounds of the BEP using gas chromatography. A significant difference was found immediately after brood capping, indicating a divergence in their pheromonal production at this time point. This is an important step to understanding the mechanisms of the Gotland population's Varroa-resistance and contributes to our global understanding of Varroa destructor infestation and survival.

Managing the parasitic honey bee mite Tropilaelaps mercedesae through combined cultural and chemical control methods.

The western honey bee (Apis mellifera) is severely impacted by the parasitic Tropilaelaps mercedesae mite, which has the capacity to outcompete Varroa destructor mites (the current leading cause of colony losses) and more rapidly overwhelm colonies. While T. mercedesae is native to Asia, it has recently expanded its geographic range and has the potential to devastate beekeeping worldwide if introduced to new regions. Our research exploited the dependence of T. mercedesae on developing honey bees (brood) by combining a cultural technique (brood break) with U.S. registered chemical products (oxalic acid or formic acid) to manage T. mercedesae infestation. To evaluate this approach, we compared four treatment groups: (1) Brood Break; (2) Brood Break + Formic Acid (FormicPro®); (3) Brood Break + Oxalic Acid dribble (Api-Bioxal®); and (4) untreated Control. We found that the mite infestation rate of worker brood in Control colonies rose from 0.4 to 15.25% over 60 days, whereas all other treatment groups had infestation rates under 0.11% on Day 60. Mite fall assessments showed similar results, whereby Control colonies had 15.48 mites fall per 24 h on day 60 compared to less than 0.2 mites for any other treatment group. Evaluation of colony strength revealed that Brood Break + Formic Acid colonies had slightly reduced adult honey bee populations. No treatment eliminated all mites, so additional measures may be needed to eradicate T. mercedesae if detected in countries that do not currently have T. mercedesae.

Pollen trapping as Honeybees pollination management and identification of dominant pollinators of Guizotia abyssinica (L.f).

Pollination is one of the most fascinating aspects of insect-plant interactions. Pollen is the male reproductive element of flowering plants, gathered by foraging Honeybees from the male parts of flowering plants called the anther. Guizotia abyssinica (L.f) is an important oilseed crop cultivated in Ethiopia and India, which belongs to the family Asteraceae. Although self-incompatibility is found in Guizotia abyssinica, a higher seed set is experienced in places with an active Honeybees population. Agricultural practices usually focus on inputs such as fertilizer application to improve seed yield of Guizotia abyssinica. However, these practices have little effect on yield if the availability of insect pollination is too low. To fill this gap, an experiment was carried out at Dandi district, West Shao zone, Oromia, Ethiopia to see the effect of Honeybees pollination management as an agronomic input. Pollination management of Honeybees was tested under feeding and non-feeding of colonies management. Pollen was trapped with and without sugar syrup feeding. The results discovered that Honeybees fed sugar syrup collected much more pollen than colonies not fed sugar syrup. The proportion of Guizotia abyssinica pollen collected through sugar syrup feeding of the colony was greater (62.2%), compared to the proportion of Guizotia abyssinica pollen trapped without sugar syrup feeding (37.8%). This indicates that sugar syrup feeding enhances the collection of pollen and probably enhances the pollination efficiency of Honeybees since they visit frequently to fulfill their daily protein requirement of pollen. Therefore, Honeybees pollination services should be included as one of the agronomic inputs with sugar syrup feeding as pollination management that might increase the yield of Guizotia abyssinica since it increases visiting frequency.

Spatiotemporal variability and foraging behavior of bee visitors to a rare long-lived iteroparous forb, Silene spaldingii (Caryophyllaceae).

Silene spaldingii S Watson is a rare long-lived forb (Caryophyllaceae) found primarily in open native grasslands of the Inland Pacific Northwest and is putatively pollinated by one key bumble bee pollinator, Bombus fervidus (Fabricius). However, populations of bumble bees and their visitation patterns can vary dramatically, and some species are in decline including B. fervidus. Understanding the role of co-pollinators such as sweat bees (Halictidae) could be crucial as the plight of rare plants and pollinators intensifies. We collected data across three seasons (2015-2017) on the Bombus-S. spaldingii pollination system, focusing on three Key Conservation Areas in the Channeled Scablands of eastern Washington. Bee visitors to S. spaldingii were monitored and the pool-of-pollinators was surveyed with blue vane traps. Nine species of bees were observed foraging on the plant, while 2211 bees comprised of five families, 22 genera and 81 taxa were captured in blue vane traps, meaning only 11.1% of species in the pollinator pool visited S. spaldingii. Halictus tripartitus Cockerell, a sweat bee, was a common visitor to the plant, but this was the first record of visitation for several other species, including Lasioglossum buccale (Pérez) which has never before been recorded in the Americas. These sweat bees appear to vector S. spaldingii pollen, suggesting they are co-pollinators of the plant. Weather and patch characteristics affected visitation patterns and the pool-of-pollinators. We conclude that sweat bees are likely co-pollinators of S. spaldingii and that they could become increasingly important if B. fervidus populations continue to decline.

Gardens reduce seasonal hunger gaps for farmland pollinators.

Gardens can benefit pollinators living in surrounding farmland landscapes, but the reason for their value is not clear. Gardens are no different from many semi-natural farmland habitats in terms of the quantity of floral resources (pollen and nectar) they produce, but the timing of their resource supply is very different, which may explain their value. We show that gardens provide 15% of overall annual nectar in farmland landscapes in Southwest UK, but between 50% and 95% during early spring and late summer when farmland supplies are low. Gardens can therefore reduce seasonal nectar gaps experienced by farmland bumblebees. Consistent with this pattern, bumblebee activity increased in gardens relative to farmland during early spring and late summer. An agent-based model reinforces this point, showing that timing, not quantity, of garden nectar supply enhances bumblebee colony growth and survival in farmland. We show that over 90% of farmland in Great Britain is within 1 km of a garden and therefore positive actions by gardeners could have widespread spillover benefits for pollinators across the country. Given the widespread distribution of gardens around the world, we highlight their important interplay with surrounding landscapes for pollinator ecology and conservation.

Stimulus intensity and temporal configuration interact during bimodal learning and memory in honey bees.

Multimodal integration is a core neural process with a keen relevance during ecological tasks requiring learning and memory, such as foraging. The benefits of learning multimodal signals imply solving whether the components come from a single event. This challenge presumably depends on the timing and intensity of the stimuli. Here, we used simultaneous and alternate presentations of olfactory and visual stimuli, at low and high intensities, to understand how temporal and intensity variations affect the learning of a bimodal stimulus and its components. We relied on the conditioning of the proboscis extension response (PER) to train honey bees to an appetitive learning task with bimodal stimuli precisely controlled. We trained bees to stimuli with different synchronicity and intensity levels. We found that synchronicity, order of presentation, and intensity significantly impacted the probability of exhibiting conditioned PER responses and the latency of the conditioned responses. At low intensities, synchronous bimodal inputs produced maximal multisensory enhancement, while asynchronous temporal orders led to lower performances. At high intensities, the relative advantage of the synchronous stimulation diminished, and asynchronous stimuli produced similar performances. Memory retention was higher for the olfactory component and bimodal stimuli compared to the visual component, irrespective of the training's temporal configuration. Bees retained the asynchronous bimodal configuration to a lesser extent than the synchronous one, depending on the stimulus intensity. We conclude that time (synchrony), order of presentation, and intensity have interdependent effects on bee learning and memory performance. This suggests caution when assessing the independent effects of each factor.

Animal pollination contributes to more than half of citrus production.

Animal pollination is crucial for the reproduction and economic viability of a wide range of crops. Despite the existing data, the extent to which citrus crops depend on pollinators to guarantee fruit production still needs to be determined. Here, we described the composition of potential pollinators in citrus (Citrus spp.) from the main growing areas of Argentina; moreover, we combined Bayesian models and empirical simulations to assess the contribution of animal pollination on fruit set and yield ha-1 in different species and cultivars of lemons, grapefruits, mandarins, and oranges. Honeybee (A. mellifera L.) was the most commonly observed potential pollinator, followed by a diverse group of insects, mainly native bees. Regardless of citrus species and cultivars, the probability of flowers setting fruit in pollinated flowers was 2.4 times higher than unpollinated flowers. Furthermore, our simulations showed that about 60% of the citrus yield ha-1 can be attributable to animal pollination across all species and cultivars. Therefore, it is crucial to maintain environments that support pollinator diversity and increase consumer and to producer awareness and demand in order to ensure the significant benefits of animal pollination in citrus production.