Intake of imidacloprid in lethal and sublethal doses alters gene expression in Apis mellifera bees.

Bees are important pollinators for ecosystems and agriculture; however, populations have suffered a decline that may be associated with several factors, including habitat loss, climate change, increased vulnerability to diseases and parasites and use of pesticides. The extensive use of neonicotinoids, including imidacloprid, as agricultural pesticides, leads to their persistence in the environment and accumulation in bees, pollen, nectar, and honey, thereby inducing deleterious effects. Forager honey bees face significant exposure to pesticide residues while searching for resources outside the hive, particularly systemic pesticides like imidacloprid. In this study, 360 Apis mellifera bees, twenty-one days old (supposed to be in the forager phase) previously marked were fed syrup (honey and water, 1:1 m/v) containing a lethal dose (0.081 µg/bee) or sublethal dose (0.00081 µg/bee) of imidacloprid. The syrup was provided in plastic troughs, with 250 µL added per trough onto each plastic Petri dish containing 5 bees (50 µL per bee). The bees were kept in the plastic Petri dishes inside an incubator, and after 1 and 4 h of ingestion, the bees were euthanised and stored in an ultra-freezer (-80 °C) for transcriptome analysis. Following the 1-h ingestion of imidacloprid, 1516 genes (73 from lethal dose; 1509 from sublethal dose) showed differential expression compared to the control, while after 4 h, 758 genes (733 from lethal dose; 25 from sublethal) exhibited differential expression compared to the control. All differentially expressed genes found in the brain tissue transcripts of forager bees were categorised based on gene ontology into functional groups encompassing biological processes, molecular functions, and cellular components. These analyses revealed that sublethal doses might be capable of altering more genes than lethal doses, potentially associated with a phenomenon known as insecticide-induced hormesis. Alterations in genes related to areas such as the immune system, nutritional metabolism, detoxification system, circadian rhythm, odour detection, foraging activity, and memory in bees were present after exposure to the pesticide. These findings underscore the detrimental effects of both lethal and sublethal doses of imidacloprid, thereby providing valuable insights for establishing public policies regarding the use of neonicotinoids, which are directly implicated in the compromised health of Apis mellifera bees.

Pequi Oil (Caryocar brasiliense Camb.) Attenuates the Adverse Effects of Cyclical Heat Stress and Modulates the Oxidative Stress-Related Genes in Broiler Chickens.

The present study was conducted to determine the possible antioxidant protection of pequi oil (PO) against cyclic heat stress in broiler chickens and to highlight the application of PO as a promising additive in broiler feed. A total of 400 one-day-old male broiler chicks (Cobb 500) were randomly assigned to 2 × 5 factorially arranged treatments: two temperature-controlled rooms (thermoneutral-TN or heat stress-HS for 8 h/day) and five dietary PO levels (0, 1.5, 3.0, 4.5, or 6.0 g/kg diet) for 42 days. Each treatment consisted of eight replicates of five birds. The results showed that HS increased glucose (p = 0.006), triglycerides (p < 0.001), and HDL (p = 0.042) at 21 days and reduced (p = 0.005) serum total cholesterol at 42 days. The results also showed that HS increased the contents of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). In contrast, PO linearly decreased AST (p = 0.048) and ALT (p = 0.020) at 21 and 42 days, respectively. The heterophil-to-lymphocyte ratio in the birds under HS was higher than in those in the TN environment (p = 0.046). Heat stress decreased (p = 0.032) the relative weight of their livers at 21 days. The superoxide dismutase activity increased (p = 0.010) in the HS treatments in comparison to the TN treatments, while the glutathione peroxidase activity in the liver decreased (p < 0.001) at 42 days; however, the activity of catalase had no significant effects. Meanwhile, increasing the dietary PO levels linearly decreased plasma malondialdehyde (p < 0.001) in the birds in the HS environment. In addition, PO reduced (p = 0.027) the expression of Hsp 70 in the liver by 92% when compared to the TN treatment without PO, mainly at the 6.0 g/kg diet level. The expression of Nrf2 was upregulated by 37% (p = 0.049) in response to PO with the 6.0 g/kg diet compared to the HS treatment without PO. In conclusion, PO supplementation alleviated the adverse effects of HS on broilers due to its antioxidant action and modulation of the genes related to oxidative stress, providing insights into its application as a potential feed additive in broiler production.

Zinc supplementation modifies brain tissue transcriptome of Apis mellifera honeybees.

BACKGROUND: Bees are the most important group of pollinators worldwide and their populations are declining. In natural conditions, Apis mellifera depends exclusively on food from the field to meet its physiological demands. In the period of scarcity, available resources are insufficient and artificial supplementation becomes essential for maintaining the levels of vitamins, proteins, carbohydrates, and minerals of colonies. Among these minerals, zinc is essential in all living systems, particularly for the regulation of cell division and protein synthesis, and is a component of more than 200 metalloenzymes. RESULTS: The total RNA extracted from the brain tissue of nurse bees exposed to different sources and concentrations of zinc was sequenced. A total of 1,172 genes in the treatment that received an inorganic source of zinc and 502 genes that received an organic source of zinc were found to be differentially expressed among the control group. Gene ontology enrichment showed that zinc can modulate important biological processes such as nutrient metabolism and the molting process. CONCLUSIONS: Our results indicate that zinc supplementation modulates the expression of many differentially expressed genes and plays an important role in the development of Apis mellifera bees. All the information obtained in this study can contribute to future research in the field of bee nutrigenomics.

Nyssorhynchus darlingi genome-wide studies related to microgeographic dispersion and blood-seeking behavior.

BACKGROUND: In Brazil, malaria is concentrated in the Amazon Basin, where more than 99% of the annual cases are reported. The main goal of this study was to investigate the population structure and genetic association of the biting behavior of Nyssorhynchus (also known as Anopheles) darlingi, the major malaria vector in the Amazon region of Brazil, using low-coverage genomic sequencing data. METHODS: Samples were collected in the municipality of Mâncio Lima, Acre state, Brazil between 2016 and 2017. Different approaches using genotype imputation and no gene imputation for data treatment and low-coverage sequencing genotyping were performed. After the samples were genotyped, population stratification analysis was performed. RESULTS: Weak but statistically significant stratification signatures were identified between subpopulations separated by distances of approximately 2-3 km. Genome-wide association studies (GWAS) were performed to compare indoor/outdoor biting behavior and blood-seeking at dusk/dawn. A statistically significant association was observed between biting behavior and single nucleotide polymorphism (SNP) markers adjacent to the gene associated with cytochrome P450 (CYP) 4H14, which is associated with insecticide resistance. A statistically significant association between blood-seeking periodicity and SNP markers adjacent to genes associated with the circadian cycle was also observed. CONCLUSION: The data presented here suggest that low-coverage whole-genome sequencing with adequate processing is a powerful tool to genetically characterize vector populations at a microgeographic scale in malaria transmission areas, as well as for use in GWAS. Female mosquitoes entering houses to take a blood meal may be related to a specific CYP4H14 allele, and female timing of blood-seeking is related to circadian rhythm genes.

Energetic supplementation for maintenance or development of Apis mellifera L. colonies.

BACKGROUND: The nutritional requirements of honeybees (Apis mellifera) for their complete development need to be supplied through food sources available in the environment, since honeybees are insects that depend directly on blossoming food sources. However, at certain times a food-supply reduction can promote nutritional stress, thus necessitating food supplementation for maintenance or production stimulus of the colonies. Thus, the determination of optimal energy supplementation can assist in the maintenance and production of colonies. METHODS: Twenty Apis mellifera beehives were used (with five beehives per treatment): CTL, control (without feeding); SJ, sugarcane juice; SS, sugar syrup; and IS, inverted sucrose. We evaluated the food consumption, population development, and physiological state (expression of vitellogenin and hexamerin 70a genes) of each colony. RESULTS: The results showed that the supplementation of colonies with sugar syrup resulted in an intermediate consumption level (894.6 ± 291 mL) and better development (384.9 ± 237.3 and 158.3 ± 171.6 cm2, sealed and open brood, respectively). Furthermore, this diet ensured that the colonies were in a good physiological state, as bees fed this diet presented the highest relative expression levels of vitellogenin and hexamerin 70a among all the diets tested. CONCLUSIONS: Therefore, sugar syrup is concluded to be the best artificial energetic food for use in the supplementation of honeybee colonies.

Energetic supplementation for maintenance or development of Apis mellifera L. colonies

The nutritional requirements of honeybees (Apis mellifera) for their complete development need to be supplied through food sources available in the environment, since honeybees are insects that depend directly on blossoming food sources. However, at certain times a food-supply reduction can promote nutritional stress, thus necessitating food supplementation for maintenance or production stimulus of the colonies. Thus, the determination of optimal energy supplementation can assist in the maintenance and production of colonies. Methods: Twenty Apis mellifera beehives were used (with five beehives per treatment): CTL, control (without feeding); SJ, sugarcane juice; SS, sugar syrup; and IS, inverted sucrose. We evaluated the food consumption, population development, and physiological state (expression of vitellogenin and hexamerin 70a genes) of each colony. Results: The results showed that the supplementation of colonies with sugar syrup resulted in an intermediate consumption level (894.6 ± 291 mL) and better development (384.9 ± 237.3 and 158.3 ± 171.6 cm2, sealed and open brood, respectively). Furthermore, this diet ensured that the colonies were in a good physiological state, as bees fed this diet presented the highest relative expression levels of vitellogenin and hexamerin 70a among all the diets tested. Conclusions: Therefore, sugar syrup is concluded to be the best artificial energetic food for use in the supplementation of honeybee colonies.(AU)

Nutritional and mineral contents of honey extracted by centrifugation and pressed processes.

In this study, wild honey samples extracted by two different methods (centrifugation and pressed processing) were characterized and compared based on their physicochemical, and nutritional properties, macro- and micro-mineral contents, and pollen counts. Twelve colonies of Africanized Apis mellifera were used; six honey samples were obtained by centrifugation and six by honeycomb press. All physicochemical parameters of honey samples (moisture, pH, total acidity, ash, dry matter, and qualitative absence of hydroxymethylfurfural) were within the limits established by EU legislation, and all parameters in pressed honey were superior (p<0.05). Nutritional contents (total carbohydrates, total lipids, total proteins, flavonoids, and ascorbic acid) and minerals (K, Ca, Mg, Na, Fe, Li, Zn) were also higher in pressed honey. The quantity of pollen in pressed honey samples was 5.6-fold higher than in centrifuged samples. Pressed honey, can be marked as a differentiated product with a higher mineral content and several nutritional properties.

Characterization of Coffea arabica monofloral honey from Espírito Santo, Brazil.

In this study, samples of coffee honey produced in Espírito Santo State, Brazil, were characterized based on their melissopalynology, physicochemical and nutritional properties, and mineral and caffeine contents. The caffeine content in the nectar from coffee flowers was measured by high performance liquid chromatography (HPLC). Blends of honey were obtained from three Coffea arabica crops, each with 10 colonies of Africanized Apis mellifera. All honey samples contained monofloral (75-78%) pollen belonging to C. arabica. Physicochemical parameters (total acidity, pH, moisture, dry matter, ash, and qualitative hydroxymethylfurfural) were within the approved limits established by EU legislation. Coffee honey contains high levels of ascorbic acid (294.68 mg kg(-1)) and low amounts of total flavonoids (3.51 ± 0.18 mg QE kg(-1)). The most abundant minerals were potassium and calcium (962.59 ± 154.3 and 343.75 ± 25.56 mg kg(-1), respectively). The caffeine content in coffee nectar (1.64 mg kg(-1)) was approximately 8-fold lower than that in honey (12.02 ± 0.81 mg kg(-1)).

Fipronil promotes motor and behavioral changes in honey bees (Apis mellifera) and affects the development of colonies exposed to sublethal doses.

Bees play a crucial role in pollination and generate honey and other hive products; therefore, their worldwide decline is cause for concern. New broad-spectrum systemic insecticides such as fipronil can harm bees and their use has been discussed as a potential threat to bees' survival. In the present study, the authors evaluate the in vitro toxicity of fipronil and note behavioral and motor activity changes in Africanized adult Apis mellifera that ingest or come into contact with lethal or sublethal doses of fipronil. The effects of sublethal doses on brood viability, population growth, behavior, and the expression of the defensin 1 gene in adult bees were studied in colonies fed with contaminated sugar syrup (8 µg fipronil L(-1) ). Fipronil is highly toxic to bees triggering agitation, seizures, tremors, and paralysis. Bees that are exposed to a lethal or sublethal doses showed reduced motor activity. The number of eggs that hatched, the area occupied by worker eggs, and the number of larvae and pupae that developed were reduced, adult bees showed lethargy, and colonies were abandoned when they were exposed to sublethal doses of fipronil. No change was seen in the bees' expression of defensin 1. The authors conclude that fipronil is highly toxic to honey bees and even sublethal doses may negatively affect the development and maintenance of colonies.

Africanized honeybees in urban areas: a public health concern.

INTRODUCTION: This study aimed to investigate the occurrence of Africanized honeybees in Botucatu, São Paulo, Brazil, and to implement a program to remove such swarms. METHODS: The occurrences of Africanized honeybee swarms between 2010 and 2012 were studied and strategies to prevent accidents were developed. RESULTS: We noted 1,164 cases of Africanized honeybee occurrences in the city, and 422 swarms were collected. The developed strategies to prevent accidents were disseminated to the population. CONCLUSIONS: We contributed to reducing the risks represented by Africanized honeybee swarms in urban areas, by collecting swarms and disseminating strategic information for preventing accidents.