Search results for: honeybees

Authors: Jacqueline Kunzelman, Val Anderson, Robert Johnson, Nicholas Anderson, Rebecca Bates

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In an effort to protect honeybees from colony collapse disorder, beekeepers are filing for government permits to use natural lands as summer pasture for honeybees under the multiple-use management regime in the United States. Utilizing natural landscapes in high mountain ranges may help strengthen honeybee colonies, as this natural setting is generally void of chemical pollutants and pesticides that are found in agricultural and urban settings. However, the introduction of a competitive species could greatly impact the native species occupying these natural landscapes. While honeybees and butterflies have different life histories, behavior, and foraging strategies, they compete for the same nectar resources. Few, if any, studies have focused on the potential population effects of commercial honeybees on native butterfly abundance and diversity. This study attempts to observe this impact using a paired before-after control-impact (BACI) design. Over the course of two years, malaise trap samples were collected every week during the months of the flowering season in two similar areas separated by 11 kilometers. Each area contained nine malaise trap sites for replication. In the first year, samples were taken to analyze and establish trends within the pollinating communities. In the second year, honeybees were introduced to only one of the two areas, and a change in trends between the two areas was assessed. Contrary to the original hypothesis, the resulting observation was an overall significant increase in the mean butterfly abundance in the impact areas after honeybees were introduced, while control areas remained relatively stable. This overall increase in abundance over the season can be attributed to an increase in butterflies during the first and second periods of the data collection when populations were near their peak. Several potential theories are 1) Honeybees are deterring a natural predator/competitor of butterflies that previously limited population growth. 2) Honeybees are consuming resources regularly used by butterflies, which may extend the foraging time and consequent capture rates of butterflies. 3) Environmental factors such as number of rainy days were inconsistent between control and impact areas, biasing capture rates. This ongoing research will help determine the suitability of high mountain ranges for the summer pasturing of honeybees and the population impacts on many different pollinators.

Keywords: butterfly, competition, honeybee, pollinator

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Authors: Fazal Said, Mian Inayatullah

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Various insect visitors in common and honeybees in particular are considered to be accountable for 80-85% of pollination services for numerous crops worldwide. Pollinators not only increase crop yield but also improve quality of produce as well. The present investigation is therefore, an endeavor to assess the visitation pattern of honeybees, Apis florea (Hymenopterae: Apidae) in sunflower (Helianthus annuus L.). The current research trial was carried out at New Developmental Farm (NDF), The University of Agriculture Peshawar, (34.01° N, 71.53° E) Khyber Pakhtunkhwa-Pakistan during 2012 and 2013. Different observations on the foraging behavior of A. florea’s individuals were made from 0800 hr in the morning and continued until 1800 hr in the evening. Hence, total duration of foraging activity of A. florea individuals was comprised of 10 hours. It was found that two peaks of visitation/foraging occurred between 1400 to 1600 hr of the day. First peak of foraging was recorded at 1600hr, where 15 individuals of honeybees/3 m2 were counted to be engaged in foraging sunflower blooms. Second peak visitation was recorded with a total of 12 bees/3 m2 at 1400 hrs of the day. Visitations of A. florea were observed to its minimum intensity of only 07 individuals during late hours of the day as evening approached after 1800 hrs. Similarly, due to more number of pollens and nectars on flowers, high frequency of A. florea were found engaged in foraging during 20th and 25th day after initiation of blooms on sunflower. Minimum numbers of honeybees were recorded during initial and very last days of flowering due to less number of plants with blooms and less availability of pollen and nectar on flowers.

Keywords: apis florea, days after flowering, daily hours, sunflower, visitation pattern

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Authors: Chakib Jerry, Mounir Jerry

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In this paper, a mathematical model of infested honey bees colonies is formulated in order to investigate Colony Collapse Disorder in a honeybee colony. CCD, as it is known, is a major problem on honeybee farms because of the massive decline in colony numbers. We introduce to the model a control variable which represents forager protection. We study the controlled model to derive conditions under which the bee colony can fight off epidemic. Secondly we study the problem of minimizing prevention cost under model’s dynamics constraints.

Keywords: honey bee, disease transmission model, disease control honeybees, optimal control

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Authors: Chin-Yuan Hsu, Yu-Lung Chuang

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The expression, concentration, and activity of mitochondrial energy-utilized molecules and cellular energy-regulated molecules decreased with age in the trophocytes and oenocytes of honeybees (Apis mellifera), but those of cellular energy-metabolized molecules is unknown. In this study, the expression, concentration, and activity of cellular energy-metabolized molecules were assayed in the trophocytes and fat cells of young and old worker bees by using the techniques of cell and biochemistry. The results showed that (i) the •-hydroxylacyl-coenzyme A dehydrogenase (HOAD) activity/citrate synthase (CS) activity ratio, non-esterified fatty acids concentrations, the expression of eukaryotic initiation factor 4E, and the expression of phosphorylated eIF4E binding protein 1 decreased with age; (ii) fat and glycogen accumulation increased with age; and (iii) the pyruvate dehydrogenase (PDH) activity/citrate synthase (CS) activity ratio was not correlated with age. These finding indicated that •-oxidation (HOAD/CS) and protein synthsis decreased with age. Glycolysis (PDH/CS) was unchanged with age. The most likely reason is that sugars are the vital food of worker bees. Taken together these data reveal that young workers have higher cellular energy metabolism than old workers and that aging results in a decline in the cellular energy metabolism in worker honeybees.

Keywords: aging, energy, honeybee, metabolism

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Authors: Chia-Ying Yen, Chin-Yuan Hsu

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The lifespans of queen honeybees (Apis mellifera) are much longer than those of worker bees. The expression, concentration, and activity of mitochondrial energy-utilized molecules decreased with age in the trophocytes and oenocytes of worker bees, but they are unknown in queen bees. In this study, the expression, concentration, and activity of mitochondrial energy-utilized molecules were evaluated in the trophocytes and oenocytes of young and old queen bees by biochemical techniques. The results showed that mitochondrial density and mitochondrial membrane potential; nicotinamide adenine dinucleotide (NAD+), nicotinamide adenine dinucleotide reduced form (NADH), and adenosine triphosphate (ATP) levels; the NAD+/NADH ratio; and relative expression of NADH dehydrogenase 1 and ATP synthase normalized against mitochondrial density were not significantly different between young and old queen bees. These findings reveal that mitochondrial energy utilization maintains a young status in the trophocytes and oenocytes of old queen bees and that trophocytes and oenocytes have aging-delaying mechanisms and can be used to study cellular longevity.

Keywords: aging, longevity, mitochondrial energy, queen bees

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Authors: Justin Orimisan Ijigbade

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The study was conducted to assess the effect of climate variability on honeybee’s production in Ondo State, Nigeria. Multistage sampling technique was employed to collect the data from 60 beekeepers across six Local Government Areas in Ondo State. Data collected were subjected to descriptive statistics and multiple regression model analyses. The results showed that 93.33% of the respondents were male with 80% above 40 years of age. Majority of the respondents (96.67%) had formal education and 90% produced honey for commercial purpose. The result revealed that 90% of the respondents admitted that low temperature as a result of long hours/period of rainfall affected the foraging efficiency of the worker bees, 73.33% claimed that long period of low humidity resulted in low level of nectar flow, while 70% submitted that high temperature resulted in improper composition of workers, dunes and queen in the hive colony. The result of multiple regression showed that beekeepers’ experience, educational level, access to climate information, temperature and rainfall were the main factors affecting honey bees production in the study area. Therefore, beekeepers should be given more education on climate variability and its adaptive strategies towards ensuring better honeybees production in the study area.

Keywords: climate variability, honeybees production, humidity, rainfall and temperature

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Authors: Nicholas Anderson, Steven Petersen, Tom Bates, Val Anderson

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Under the multiple-use management regime established in the United States for federally owned lands, government agencies have come under pressure from commercial apiaries to grant permits for the summer pasturing of honeybees on government lands. Federal agencies have struggled to integrate honeybees into their management plans and have little information to make regulations that resolve how many colonies should be allowed in a single location and at what distance sets of hives should be placed. Many conservation groups have voiced their concerns regarding the introduction of honeybees to these natural lands, as they may outcompete and displace native pollinating species. Assessing the quality of an area in regard to its floral resources, pollen, and nectar can be important when attempting to create regulations for the integration of commercial honeybee operations into a native ecosystem. Areas with greater floral resources may be able to support larger numbers of honeybee colonies, while poorer resource areas may be less resilient to introduced disturbances. Attempts are made in this study to determine flower cover using high resolution drone imagery to help assess the floral resource availability to pollinators in high elevation, tall forb communities. This knowledge will help in determining the potential that different areas may have for honeybee pasturing and honey production. Roughly 700 images were captured at 23m above ground level using a drone equipped with a Sony QX1 RGB 20-megapixel camera. These images were stitched together using Pix4D, resulting in a 60m diameter high-resolution mosaic of a tall forb meadow. Using the program ENVI, a supervised maximum likelihood classification was conducted to calculate the percentage of total flower cover and flower cover by color (blue, white, and yellow). A complete vegetation inventory was taken on site, and the major flowers contributing to each color class were noted. An accuracy assessment was performed on the classification yielding an 89% overall accuracy and a Kappa Statistic of 0.855. With this level of accuracy, drones provide an affordable and time efficient method for the assessment of floral cover in large areas. The proximal step of this project will now be to determine the average pollen and nectar loads carried by each flower species. The addition of this knowledge will result in a quantifiable method of measuring pollen and nectar resources of entire landscapes. This information will not only help land managers determine stocking rates for honeybees on public lands but also has applications in the agricultural setting, aiding producers in the determination of the number of honeybee colonies necessary for proper pollination of fruit and nut crops.

Keywords: honeybee, flower, pollinator, remote sensing

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Authors: Dani Sukkar, Ali Kanso, Philippe Laval-Gilly, Jairo Falla-Angel

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The honeybees' immune system is challenged by different risk factors that induce various responses. However, complex scenarios where bees are exposed to different pesticides simultaneously with immune activation are not well evaluated. The Toll pathway is one of the main signaling pathways studied in invertebrate immune responses, and it is a good indicator of the effect of such complex interactions in addition to key signaling elements of other pathways like Relish of the immune deficiency (IMD) pathway or Eater, the phagocytosis receptor or vitellogenin levels. Honeybee hemocytes extracted from 5th instar larvae were exposed to imidacloprid and/or amitraz with or without the presence of the zymosan a as an immune activator. The gene expression of multiple immune related genes were studied, including spaetzle, Toll, myD88, relish, eater and vitellogenin, by real-time polymerase chain reaction after RNA extraction. The results demonstrated that the Toll pathway is mainly affected by the pesticides; imidacloprid and amitraz, especially by their different combinations. Furthermore, immune activation by zymosan A, a fungal cell-wall component, acts to mitigate to some extent the effect of pesticides on the different levels of the Toll pathway. In addition, imidacloprid, amitraz, and zymosan A have complex and context-specific interactions depending on the levels of immune activation and the pathway evaluated affecting immune-gene expression differently.

Keywords: toll pathway, immune modulation, β-glucan, imidacloprid, amitraz, honeybees, immune genes

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Authors: John Michael Russo

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Honeybees are important to our food system and continue to suffer from high rates of colony loss. Precision feed has brought many benefits to livestock cultivation and these should transfer to apiculture. However, apiculture has unique challenges. The objective of this research is to understand how principles of precision agriculture, applied to apiculture and feed specifically, might effectively improve state-of-the-art cultivation. The methodology surveys apicultural practice to build a model for assessment. First, a review of apicultural motivators is made. Feed method is then evaluated. Finally, precision feed methods are examined as accelerants with potential to advance the effectiveness of feed practice. Six important motivators emerge: colony loss, disease, climate change, site variance, operational costs, and competition. Feed practice itself is used to compensate for environmental variables. The research finds that the current state-of-the-art in apiculture feed focuses on critical challenges in the management of feed schedules which satisfy requirements of the bees, preserve potency, optimize environmental variables, and manage costs. Many of the challenges are most acute when feed is used to dispense medication. Technology such as RNA treatments have even more rigorous demands. Precision feed solutions focus on strategies which accommodate specific needs of individual livestock. A major component is data; they integrate precise data with methods that respond to individual needs. There is enormous opportunity for precision feed to improve apiculture through the integration of precision data with policies to translate data into optimized action in the apiary, particularly through automation.

Keywords: precision agriculture, precision feed, apiculture, honeybees

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Authors: Alebiosu Olugbenga Shadrak, Victor Victoria

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Honey bees exhibit preferences in their foraging behaviour on pollen and nectar for food and honey production, respectively. Melissopalynology is the study of pollen in honey and other honey products. Several work have been conducted on the palynological studies of honeys from the southern parts of Nigeria but with relatively scant records from the Northern region of the country. This present study aimed at revealing the favourably visited plants by honey bees, Apis melifera var. adansonii, at some apiaries in Northern Nigeria, as well as determining the quality of honeys produced. Honeys were harvested and collected from four apiaries of the region, namely: Sarkin Dawa missionary bee farm, Taraba State; Eleeshuwa Bee Farm, Keffi, Nassarawa State, Bulus Beekeeper Apiaries, Kagarko, Kaduna State and Mai Gwava Bee Farm, Kano State. These honeys were acetolysed for palynological microscopic analysis and subjected to standard treatment methods for the determination of their proximate composition and sugar profiling. Fresh anthers of two dominantly represented plants in the honeys were then collected for the quantification of their pollen protein contents, using the micro-kjeldhal procedure. A total of 30 pollen types were identified in the four honeys, and some of them were common to the honeys. A classification method for expressing pollen frequency class was employed: Senna cf. siamea, Terminalia cf. catappa, Mangifera indica, Parinari curatelifolia, Vitellaria paradoxa, Elaeis guineensis, Parkia biglobosa, Phyllantus muellerianus and Berlina Grandiflora, as “Frequent” (16-45%); while the others are either Rare (3-15%) or Sporadic (less than 3 %). Pollen protein levels of the two abundantly represented plants, Senna siamea (15.90mg/ml) and Terminalia catappa (17.33mg/ml) were found to be considerably lower. The biochemical analyses revealed varying amounts of proximate composition, non-reducing sugar and total sugar levels in the honeys. The results of this study indicate that pollen and nectar of the “Frequent” plants were preferentially foraged by honeybees in the apiaries. The estimated pollen protein contents of Senna same and Terminalia catappa were considerably lower and not likely to have influenced their favourable visitation by honeybees. However, a relatively higher representation of Senna cf. siamea in the pollen spectrum might have resulted from its characteristic brightly coloured and well scented flowers, aiding greater entomophily. Terminalia catappa, Mangifera indica, Elaeis guineensis, Vitellaria paradoxa, and Parkia biglobosa are typical food crops; hence they probably attracted the honeybees owing to the rich nutritional values of their fruits and seeds. Another possible reason for a greater entomophily of the favourably visited plants are certain nutritional constituents of their pollen and nectar, which were not investigated in this study. The nutritional composition of the honeys was observed to fall within the safe limits of international norms, as prescribed by Codex Alimentarius Commission, thus they are good honeys for human consumption. It is therefore imperative to adopt strategic conservation steps in ensuring that these favourably visited plants are protected from indiscriminate anthropogenic activities and also encourage apiarists in the country to establish their bee farms more proximally to the plants for optimal honey yield.

Keywords: honeybees, melissopalynology, preferentially foraged, nutritional, bee farms, proximally

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Authors: Noureddine Adjlane, Nizar Haddad

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Deformed Wing Virus (DWV) is considered as the most prevalent virus that dangerous the honeybee health worldwide today. In this study we aimed to evaluate the impact of the virus on honeybees (Apis mellifera intermissa) mortality in Algeria and we conducted the study on samples collected from the central area in the country. We used PCR for the diagnoses of the (DWV) in the diagnosis. The results had shown a high infestation in the sampled colonies and it represented 42% of the total sample. In this study, we found a clear role of both Varroa destructor mite and DWV on hive mortality in the experimented apiary. Further studies need to be conducted in order to give soled recommendations to the beekeepers, decision makers and stockholders of the Algerian beekeeping sector.

Keywords: honey bee, DWV, Varroa destructor, mortality, prevalence, infestation

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Authors: Erol Bayraktarov, Asya Ilgun, Thomas Schickl, Alexandre Campo, Nicolis Stamatios

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Traditional means of harvesting honey are often stressful for honeybees. Each time honey is collected a portion of the colony can die. In consequence, the colonies’ resilience to environmental stressors will decrease and this ultimately contributes to the global problem of honeybee colony losses. As part of the project HIVEOPOLIS, we design and build a different kind of beehive, incorporating technology to reduce negative impacts of beekeeping procedures, including honey harvesting. A first step in maintaining more sustainable honey harvesting practices is to design honey storage frames that can automate the honey collection procedures. This way, beekeepers save time, money, and labor by not having to open the hive and remove frames, and the honeybees' nest stays undisturbed.This system shows promising features, e.g., high reliability which could be a key advantage compared to current honey harvesting technologies.Our original concept of fractional honey harvesting has been to encourage the removal of honey only from "safe" locations and at levels that would leave the bees enough high-nutritional-value honey. In this abstract, we describe the current state of our honey harvester, its technology and areas to improve. The honey harvester works by separating the honeycomb cells away from the comb foundation; the movement and the elastic nature of honey supports this functionality. The honey sticks to the foundation, because of the surface tension forces amplified by the geometry. In the future, by monitoring the weight and therefore the capped honey cells on our honey harvester frames, we will be able to remove honey as soon as the weight measuring system reports that the comb is ready for harvesting. Higher viscosity honey or crystalized honey cause challenges in temperate locations when a smooth flow of honey is required. We use resistive heaters to soften the propolis and wax to unglue the moving parts during extraction. These heaters can also melt the honey slightly to the needed flow state. Precise control of these heaters allows us to operate the device for several purposes. We use ‘Nitinol’ springs that are activated by heat as an actuation method. Unlike conventional stepper or servo motors, which we also evaluated throughout development, the springs and heaters take up less space and reduce the overall system complexity. Honeybee acceptance was unknown until we actually inserted a device inside a hive. We not only observed bees walking on the artificial comb but also building wax, filling gaps with propolis and storing honey. This also shows that bees don’t mind living in spaces and hives built from 3D printed materials. We do not have data yet to prove that the plastic materials do not affect the chemical composition of the honey. We succeeded in automatically extracting stored honey from the device, demonstrating a useful extraction flow and overall effective operation this way.

Keywords: honey harvesting, honeybee, hiveopolis, nitinol

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Authors: Mohammed M. Ibrahim, A. A. Yusuf, Manuel Du, Fiona Mumoki

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The traditional honey bee species of Sudan is Apis mellifera, but in 1985, the dwarf bee Apis florea was introduced to the country, so now there are two species present. However, there are conflicting assessments regarding the subspecies of Apis mellifera colonies in Sudan. Likewise, it is unclear if, in the 40 years since its introduction, Apis florea has already developed regional differences or ecotypes. To shed light on these questions, we performed a morphology study on Sudanese honeybees. Samples of 10 to 20 honeybee workers per colony of the two species were collected from 16 locations, spanning different climatic zones in Sudan during 2021. Measurements were taken from 16 morphometric characteristics using a stereo-microscope equipped with an Image Analysis System (Moticam Image Plus 5.0 Digital Microscope Camera) to study their variability. The results indicate that in both species, the general means of various characters showed significant differences (p < 0.05) within a species between different locations, indicating that there might indeed be regional differences. However, more taxonomic investigation and, ideally also, molecular studies are needed in order to confirm the proper identification of subspecies and their ecotypes.

Keywords: Apis, subspecies, morphology, Sudan

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Authors: Damien P. Fevre, Peter K. Dearden

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Honeybee population sustainability is a critical concern for environmental stability and human food security. The success of a colony relies heavily on the egg-laying capacity of the queen, as it determines the production of thousands of worker bees who, in turn, perform essential functions in foraging and transforming food to make it digestible for the colony. The main sources of nutrition for honeybees are nectar, providing carbohydrates, and pollen, providing protein. This study delves into the impact of the proportion of these macronutrients on the food consumption patterns of nurse bees responsible for feeding the queen and how it affects the characteristics of the eggs produced. Using nutritional geometry, qRT-PCR, and RNA-seq analysis, this study sheds light on the pivotal role of nutrition in influencing gene expression in nurse bees, honeybee queen egg-laying capacity and embryonic development. Interestingly, while nutrition is crucial, the queen's genotype plays an even more significant role in this complex relationship, highlighting the importance of genotype-by-environment interactions. Understanding the interplay between genotype and nutrition is key to optimizing beekeeping management and strategic queen breeding practices. The findings from this study have significant implications for beekeeping practices, emphasizing the need for an appropriate nutrition to support the social nutrition of Apis mellifera. Implementing these insights can lead to improved colony health, increased productivity, and sustainable honeybee conservation efforts.

Keywords: honeybee, egg-laying, nutrition, transcriptomics

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Authors: C. V. Nnamani, O. L. Adedeji

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The contemporary global economic meltdown has devastating effect on the Nigerian’s economy and its frantic search for alternative source of national revenue aside from oil and gas has become imperative for economic emancipation for Nigerians. Apicultural entrepreneurship could provide a source of livelihood if the basic knowledge of those plant genetic resources needed by bees is made available. A palynological evaluation of those palynotaxa which honey bees forage for pollen and nectar was carried out after standard acetolysis method. Results showed that the honey samples were highly diversified and rich in honey plants. A total of 9544.3 honey pollen, consisting of 39 honey plants belonging to 21 plant families and distributed within 38 genera were identified excluding 238 unidentified pollen grains. Data from the analysis equally revealed that Elaeis guineensis Jacq, Anacardium occidentale L, Diospyros mespiliformis Hochist xe ADC, Alchornea cordifolia Muell, Arg, Daniella oliveri (Rolfe) Hutch & Dalz, Irvingia wombolu Okafor ex Baill, Treculia africana Decne, Nauclea latifolia Smith and Crossopteryx febrifuga Afzil ex Benth were the predominant honey plants. It provided a guide to the optimal utilization of floral resources by honeybees in these regions, showing the opportunity and amazing potentials for apiculture entrepreneurship of these palytaxa. Most of these plants are rare, threatened and endangered. It calls for urgent conservation techniques and step by all players. Critical awareness creation to ensure farmers knowledge of these palynotaxa to ensure proper understanding and attendance boost from them as economic empowerment is needed.

Keywords: palynotaxa, acetolysis, enterprise, livelihood, Nigeria

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Authors: Ricardo de Oliveira Orsi, Aline Astolfi, Daniel Diego Mendes, Isabella Cristina de Castro Lippi, Jaine da Luz Scheffer, Yan Souza Lima, Juliana Lunardi, Giovanna do Padro Ribeiro, Samir Moura Kadri

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NECTAR Brazilian group (Center of Education, Science, and Technology in Rational Beekeeping) conducted studies on the pesticides honey bees effects using the transcriptome sequencing (RNA-Seq) analyzes for gene expression studies. In this way, we analyzed the effects of Pyraclostrobin and Fipronil on the honey bees with 21 old-days (forager) in laboratory conditions. For this, frames containing sealed brood were removed from the beehives and maintenance on the stove (32°C and 75% humidity) until the bees were born. So, newly emerged workers were marked on the pronotum with a non-toxic pen and reintroduced into their original hives. After 21 days, 120 marked bees were collected with an entomological forces and immediately stored in Petri dishes, perforated to ensure ventilation, and kept fasted for 3 hours. These honeybees were exposed to food contaminated or not with the sublethal dose of Pyraclostrobin (850 ppb/bee) or Fipronil (2.5 ppb/bee). After four hours of exposure, 15 bees from each treatment were referred to transcriptome analysis. Total RNA analysis was extracted from the brain pools (03 brains per pool) using the TRIzol® reagent protocol according to the manufacturer's instructions. cDNA libraries were constructed, and the FASTQC program was used to check adapter content and assess the quality of raw reads. Differential expression analysis was performed with the DESeq2 package. Genes that had an adjusted value of less than 0.05 were considered to be significantly up-regulated. Regarding the Pyraclostrobin, alterations were observed in the pattern of 17 gene related to of antioxidant system, cellular respiration, glucose metabolism, and regulation of juvenile hormone and the hormone insulin. Glyphosate altered the 10 gene related to the digestive system, exoskeleton composition, vitamin E transport, and antioxidant system. The results indicate that the necessity of studies using the sublethal doses to evaluate the pesticides uses and risks on crops and its effects on the honey bees.

Keywords: beekeeping, honey bees, pesticides, transcriptome

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Authors: Ridwaan N. Milase, Leonardo J. Van Zyl, Marla Trindade

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American foulbrood (AFB) is the world’s most devastating honeybee disease that has drastically reduced the population of Apis mellifera capensis since 2009. The outbreak has jeopardized the South African bee keeping industry as well as the agricultural sector dependent on honeybees for honey production and pollination, leading to significant economic losses. AFB is caused by Paenibacillus larvae, a spore-forming, Gram positive facultative anaerobic and flagellated bacterium. The use of antibiotics within beehives has selected for resistant strains of P. larvae, while the current practice of burning spore contaminated beehives and equipment contributes to the economic losses in the honeybee-keeping industry. Therefore, phage therapy is proposed as a promising alternative to combat P. larvae strains affecting A. mellifera capensis. The genomes of two P. larvae strains isolated from infected combs in the Western Cape have been sequenced and annotated using bioinformatics tools. Genome analyses has revealed that these P. larvae strains are lysogens to more than 6 different prophages and possess different type of clustered regularly interspaced short palindromic repeat (CRISPRs) regions per strain. Active prophages from one of the two P. larvae strains were detected and identified using PCR. Electron microscopy was used to determine the family of the identified active prophages. Lytic bacteriophages that specifically target the two P. larvae strains were purified from sewage wastewater, beehive materials, and soil samples to investigate their potential development as anti-P. larvae agents. Another alternative treatment being investigated is the development of a prophage endolysin cocktail. Endolysin genes of the prophages have been targeted, cloned and expressed in Escherichia coli. The heterologously expressed endolysins have been purified and are currently being assessed for their lytic activity against P. larvae strains and other commensal microorganisms that compose the honeybee larvae microbiota. The study has shown that phage therapy and endolysins have a great potential as alternative control methods for AFB disease affecting A. mellifera capensis.

Keywords: American foulbrood, bacteriophage, honeybee, Paenibacillus larvae

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Authors: Luana Guerra, Silvio C. Sampaio, Vladimir Pavan Margarido, Ralpho R. Reis

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Brazil is the world's largest consumer of pesticides. The excessive use of these compounds has negative impacts on animal and human life, the environment, and food security. Bees, crucial for pollination, are exposed to pesticides during the collection of nectar and pollen, posing risks to their health and the food chain, including honey contamination. Aquatic sediments are also affected, impacting water quality and the microbiota. Therefore, the analysis of aquatic sediments and bee honey is essential to identify environmental contamination and monitor ecosystems. The aim of this study was to use samples of honey from honeybees (Apis mellifera) and aquatic sediment as bioindicators of environmental contamination by pesticides and their relationship with agricultural use in the surrounding areas. The sample collections of sediment and honey were carried out in two stages. The first stage was conducted in the Bituruna municipality region in the second half of the year 2022, and the second stage took place in the regions of Laranjeiras do Sul, Quedas do Iguaçu, and Nova Laranjeiras in the first half of the year 2023. In total, 10 collection points were selected, with 5 points in the first stage and 5 points in the second stage, where one sediment sample and one honey sample were collected for each point, totaling 20 samples. The honey and sediment samples were analyzed at the Laboratory of the Paraná Institute of Technology, with ten samples of honey and ten samples of sediment. The selected extraction method was QuEChERS, and the analysis of the components present in the sample was performed using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The pesticides Azoxystrobin, Epoxiconazole, Boscalid, Carbendazim, Haloxifope, Fomesafen, Fipronil, Chlorantraniliprole, Imidacloprid, and Bifenthrin were detected in the sediment samples from the study area in Laranjeiras do Sul, Paraná, with Carbendazim being the compound with the highest concentration (0.47 mg/kg). The honey samples obtained from the apiaries showed satisfactory results, as they did not show any detection or quantification of the analyzed pesticides, except for Point 9, which had the fungicide tebuconazole but with a concentration Keywords: contamination, water research, agrochemicals, beekeeping activity

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