Exploring honey bee toxicological data as a proxy for assessing dimethoate sensitivity in stingless bees.

The high diversity and distinctive characteristics of stingless bees pose challenges in utilizing toxicity test results for agrochemical registrations. Toxicity assessments were performed on 15 stingless bee species, along with the honey bee, using the insecticide dimethoate, following adapted OECD protocols. Median lethal doses over 24 h (24 h-LD50) were determined for exposure routes (acute oral or contact) and species. Species sensitivity distribution (SSD) curves were constructed and the 5% hazard doses (HD5) were estimated based on 24 h-LD50 values. The SSD curve was adjusted as the body weight and dimethoate response were correlated. Lighter bees (<10 mg) had lower 24 h-LD50 values. Contact exposure for adjusted HD5 suggested insufficient protection for Melipona mondury, whereas the oral exposure HD5 indicated no risks for the other 14 species. Comprehensive risk assessments are crucial for understanding the agrochemical impact on stingless bees, emphasizing the need for a broader species range in formulating conservation strategies.

Investigating the effects of mesotrione/atrazine-based herbicide on honey bee foragers.

A mixture of the herbicides mesotrione and atrazine (Calaris®) is a widely used herbicide in agriculture in several countries. However, the possible toxicological effects of this formulation on non-target organisms require investigation. In this study, the effects of acute oral exposure to Calaris® were evaluated in Apis mellifera foragers. The effect of seven different concentrations of Calaris® on survival and sucrose consumption was studied, while the recommended concentration for field use (FC) and its 10× dilution (0.1 FC) were used to assess overall locomotor activity, respiratory rate, flight, midgut morphology, oxidative and nitrosative stresses, and hemocyte counting. The exposure to FC or 0.1 FC decreased locomotor activity and induced damage to the midgut epithelium. Additionally, the two tested concentrations reduced superoxide dismutase activity, nitric oxide levels, and total hemocyte count. FC also increased malondialdehyde content and 0.1 FC increased respiratory rate and decreased the proportion of prohemocytes. Overall, our findings evidenced significant harmful effects on A. mellifera foragers resulting from the ingestion of the Calaris® herbicide.

Ingesting microplastics or nanometals during development harms the tropical pollinator Partamona helleri (Apinae: Meliponini).

The disposal of plastics and metal-derived compounds results in the contamination of the environment with nano/microparticles, leading to the exposure of various organisms to these harmful particles. However, the impacts of these particles on pollinating insects, which provide relevant ecosystem services, are not well understood. The aim of this study was to assess the effects of microscopic particles on the tropical pollinator Partamona helleri (Apinae: Meliponini), specifically evaluating the toxicity of plastic microparticles (polystyrene - PS, and polyethylene terephthalate - PET) and nanoparticles of a metal oxide (titanium dioxide - TiO2) via larval ingestion by bees reared in vitro. The survival rate of P. helleri larvae was not affected by the ingestion of particles of PS (500 ng/bee), PET (500 ng/bee), or TiO2 (10 µg/bee) compared to the non-treated diet (control or diet without the particles). Adults derived from treated larvae had increased body weight compared to the control, and the walking behavior of adults was altered by the ingestion of particles. Adults that ingested PET or TiO2 as larvae tended to rest for a longer time and interact more with other bees than the control. Hemocyte counts also changed, with a shift in the proportion of plasmatocytes and prohemocytes in treated individuals. Our findings suggest that even levels considered low for honey bees of exposure to plastic microparticles or metal nanoparticles can harm the health and behavior of stingless bees.

Bioinsecticide spinosad poses multiple harmful effects on foragers of Apis mellifera.

There are multifactorial causes for the recent decline in bee populations, which has resulted in compromised pollination and reduced biodiversity. Bees are considered one of the most important non-target insects affected by insecticides used in crop production. In the present study, we investigated the effects of acute oral exposure to spinosad on the survival, food consumption, flight behavior, respiration rate, activity of detoxification enzymes, total antioxidant capacity (TAC), brain morphology, and hemocyte count of Apis mellifera foragers. We tested six different concentrations of spinosad for the first two analyses, followed by LC50 (7.7 mg L-1) for other assays. Spinosad ingestion decreased survival and food consumption. Exposure to spinosad LC50 reduced flight capacity, respiration rate, and superoxide dismutase activity. Furthermore, this concentration increased glutathione S-transferase activity and the TAC of the brain. Notably, exposure to LC50 damaged mushroom bodies, reduced the total hemocyte count and granulocyte number, and increased the number of prohemocytes. These findings imply that the neurotoxin spinosad affects various crucial functions and tissues important for bee performance and that the toxic effects are complex and detrimental to individual homeostasis.

Effects of dietary electrolyte balance on performance, energy balance, and expression of genes related to acid-basic balance, absorption, and transport of nutrients in broilers.

The objective of this study was to evaluate the effect of dietary electrolyte balance (DEB) levels on performance characteristics (feed intake, FI; body weight gain, BWG; and feed efficiency, FE), energy balance (retained energy, RE; metabolizable energy ingested, MEI; heat production, HP; and energy retention efficiency, ERE), and the expression of genes related to acid-base balance, nutrient absorption, and transport in broilers from 1 to 21 days of age. A total of 245 male Cobb chickens were used in a completely randomized design with five DEB levels (110, 175, 240, 305, and 370 mEq/kg) and seven replicates of seven birds each. The inclusion of DEB levels influenced FE; 110 mEq/kg provided the better values for this characteristic both in the pre-initial phase and in the initial phase but was different only concerning 175 mEq/kg (1-7 days) and 240 mEq/kg (1-21 days). Birds that ingested diets with a level of 240 mEq/kg of DEB had a higher MEI and HP. This DEB level caused a lesser and greater expression of the SLC12A2 gene in the liver and the ATP1A1 gene in the intestine, respectively. On the other hand, the ATP1A1 gene was less expressed in the liver and kidney of broilers supplemented with 370 mEq/kg compared to a level of 110 mEq/kg. In general, a level of 110 mEq/kg DEB in the ration seems to be the most suitable for good performance, energy balance, and gene expression of broilers from 1 to 21 days of age.

Selection of reference genes for quantitative real-time PCR normalization in European quail tissues.

Coturniculture has been standing out as an industrial poultry activity in several countries around the world because of the several adaptive advantages of quails. Research that considers the analysis of gene expression can enhance this activity. This study aimed to analyze the stability of reference genes (RGs) in different tissues of quails (both males and females) for the recommendation of use in gene expression studies by the quantitative reverse transcription-polymerase chain reaction (RT-qPCR). The expression stability of ten RGs (ACTA1, ACTB, B2M, GAPDH, HMBS, SDHA, HPRT1, MRPS27, MRPS30, and RPL5) was analyzed in four tissues (breast muscle, abdominal fat, liver, and intestine), and assessed using the statistical tools geNorm, NormFinder, comparative ΔCq method, and BestKeeper. The HPRT1 gene was the most stable in all quail tissues tested, followed by MRPS27 and MRPS30 in breast muscle, B2M and RPL5 in abdominal fat, HMBS and B2M in the liver, and RPL5 and HMBS in the intestine. These results may help studies using RT-qPCR assays to assess quail tissues from both sexes because they provide data on the most stable genes, which should be tested as candidate RGs for other experimental conditions.

A mixture containing the herbicides Mesotrione and Atrazine imposes toxicological risks on workers of Partamona helleri.

A mixture of Mesotrione and Atrazine (Calaris®) has been reported as an improvement of the atrazine herbicides, which are agrochemicals used for weed control. However, its possible harmful effects on non-target organisms, including pollinators, needs to be better understood. In this work, the effects of the mix of herbicides on food consumption, behaviour (walking distance, and meandering), and the morphology of the midgut of the stingless bee Partamona helleri were studied. Foragers were orally exposed to different concentrations of the mix. The concentrations leading to 10% and 50% mortality (LC10 and LC50, respectively) were estimated and used in the analysis of behaviour and morphology. The ingestion of contaminated diets (50% aqueous sucrose solution + mix) led to a reduction in food consumption by the bees when compared to the control, bees fed a non-contaminated diet (sucrose solution). Ingestion of the LC50 diet reduced locomotor activity, increased meandering, induced the degradation of the epithelium and peritrophic matrix, and also changed the number of cells positive for signalling-pathway proteins in the midgut. These results show the potential toxicological effects and environmental impacts of the mix of herbicides in beneficial insects, including a native bee.

Spinosad-mediated effects in the post-embryonic development of Partamona helleri (Hymenoptera: Apidae: Meliponini).

The use of insecticides based on metabolites found in live organisms, such as the insecticide spinosad, has been an option for the control of agricultural pests because of the allegedly low toxicological risk for nontarget arthropods, such as stingless bees. In the current study, we evaluate the effects of chronic oral exposure to spinosad during the larval phase on survival, developmental time, body mass, midgut epithelial remodeling, and the peritrophic matrix (PM) of Partamona helleri stingless bee workers. Worker larvae that were raised in the laboratory were orally exposed to different concentrations (0, 6.53, 13.06, 32.64, and 3,264 ng. a.i. bee-1) of spinosad (formulation), and the resulting survival, developmental time, and body mass were studied. The concentration of spinosad recommended for use in the field (3,264 ng. a.i. bee-1) reduced the survival of workers during development. Also, sublethal concentrations of spinosad delayed the development and caused morphological changes in the midgut epithelium. Finally, the chronic exposure of larvae to 32.64 ng. a.i. bee-1 spinosad also altered the remodeling of the midgut during metamorphosis and affected the organization of the PM of larvae, pupae, and adults. Our data suggest possible environmental risks for using spinosad in cultures that are naturally pollinated by stingless bees.

Spinosad-mediated effects on survival, overall group activity and the midgut of workers of Partamona helleri (Hymenoptera: Apidae).

Populations of stingless bees have declined around the world and pesticides have been indicated as one of the possible causes of this decrease. Spinosad, which is synthesized from the fermentation process produced by the soil actinomycete Saccharopolyspora spinosa, is one of the most used bioinsecticides today. This study aimed to evaluate the possible effects of spinosad (formulation) on survival, general group activity and the processes of autophagy, apoptosis and oxidative stress in two organs (midgut and brain) of workers of Partamona helleri, after 24 h of oral exposure. Workers were orally exposed to different concentrations of spinosad. The concentration (8.16 × 10-3 mg a.i./mL) that led to the mortality of approximately half the number of treated bees was considered LC50 and was used in behavior, histology and immunofluorescence bioassays. The results revealed that bee survival was substantially reduced with increasing spinosad concentrations. The LC50 of the bioinsecticide compromised general group activity, caused morphological alterations in the midgut and intensified the processes of autophagy, apoptosis and oxidative stress in this organ. The brain, on the other hand, did not present significant alterations under the tested conditions. The data obtained demonstrate, therefore, that spinosad negatively affects individual survival, general group activity and the midgut epithelium of P. helleri.