Bixafen, Prothioconazole, and Trifloxystrobin Alone or in Combination Have a Greater Effect on Health Related Gene Expression in Honey Bees from Nutritionally Deprived than from Protein Supplemented Colonies.

The aim of this study was to evaluate whether alterations in food availability compromise the metabolic homeostasis of honey bees exposed to three fungicides alone or together. Ten honey bee colonies were used, with half receiving carbohydrate-protein supplementation for 15 weeks while another five colonies had their protein supply reduced with pollen traps. Subsequently, forager bees were collected and exposed by contact to 1 or 7 µg of bixafen, prothioconazole, or trifloxystrobin, either individually or in combination. After 48 h, bee abdomens without the intestine were used for the analysis of expression of antioxidant genes (SOD-1, CAT, and GPX-1), detoxification genes (GST-1 and CYP306A1), the storage protein gene vitellogenin, and immune system antimicrobial peptide genes (defensin-1, abaecin, hymenoptaecin, and apidaecin), through real-time PCR. All fungicide treatments induced changes in gene expression, with bixafen showing the most prominent upregulation. Exposure to 1 µg of each of the three pesticides resulted in upregulation of genes associated with detoxification and nutrition processes, and downregulation of immune system genes. When the three pesticides were combined at a dose of 7 µg each, there was a pronounced downregulation of all genes. Food availability in the colonies affected the impact of fungicides on the expression of the studied genes in forager bees.

Mitochondrial Respiratory Inhibition Promoted by Pyraclostrobin in Fungi is Also Observed in Honey Bees.

There is no use restriction associated with bees for many fungicides used in agriculture; however, this does not always mean that these pesticides are harmless for these nontarget organisms. We investigated whether the fungicide pyraclostrobin, which acts on fungal mitochondria, also negatively affects honey bee mitochondrial bioenergetics. Honey bees were collected from 5 hives and anesthetized at 4 °C. The thoraces were separated, and mitochondria were isolated by grinding, filtering, and differential centrifugation. An aliquot of 0.5 mg of mitochondrial proteins was added to 0.5 mL of a standard reaction medium with 4 mM succinate (complex II substrate) plus 50 nM rotenone (complex I inhibitor), and mitochondrial respiration was measured at 30 °C using a Clark-type oxygen electrode. Mitochondrial membrane potential was determined spectrofluorimetrically using safranin O as a probe, and adenosine triphosphate (ATP) synthesis was determined by chemiluminescence. Pyraclostrobin at 0 to 50 µM was tested on the mitochondrial preparations, with 3 repetitions. Pyraclostrobin inhibited mitochondrial respiration in a dose-dependent manner at concentrations of 10 µM and above, demonstrating typical inhibition of oxidative phosphorylation. Pyraclostrobin also promoted a decline in the mitochondrial membrane potential at doses of 5 µM and above and in ATP synthesis at 15 µM and above. We conclude that pyraclostrobin interferes with honey bee mitochondrial function, which is especially critical for the energy-demanding flight activity of foraging bees. Environ Toxicol Chem 2020;39:1267-1272. © 2020 SETAC.

Pyraclostrobin Impairs Energetic Mitochondrial Metabolism and Productive Performance of Silkworm (Lepidoptera: Bombycidae) Caterpillars.

Silkworm cocoon production has been reduced due to a number of problems other than those inherent in sericulture, such as diseases, malnutrition, and inappropriate management. The use of pesticides in areas surrounding mulberry fields can contaminate these plants and consequently harm caterpillars. The aim of this study was to evaluate whether the application of the fungicide pyraclostrobin in mulberry plants interferes with the mitochondrial bioenergetics and the productive performance of silkworms. Mulberry plants were treated with pyraclostrobin (0, 100, 200, and 300 g ha-1). After 30 d of fungicide application, fifth instar caterpillars were fed with leaves from the treated plants. We evaluated in vitro and in vivo mitochondrial bioenergetics of mitochondria from the head and intestines, as well as the feed intake and mortality rate of the caterpillars and the weight of fresh cocoons and cocoons shells. At doses of 50 µM (in vitro) and 200 g ha-1 (in vivo), pyraclostrobin inhibited oxygen consumption in state 3, dissipated membrane potential, and inhibited ATP synthesis in mitochondria. Pyraclostrobin acted as a respiratory chain inhibitor, affecting mitochondrial bioenergetics. The fungicide did not interfere with food consumption but negatively affected mortality rate and weight of cocoons. Mulberry leaves contaminated with pyraclostrobin negatively impact the mitochondrial bioenergetics of silkworms and cocoon production.

Fipronil and imidacloprid reduce honeybee mitochondrial activity.

Bees have a crucial role in pollination; therefore, it is important to determine the causes of their recent decline. Fipronil and imidacloprid are insecticides used worldwide to eliminate or control insect pests. Because they are broad-spectrum insecticides, they can also affect honeybees. Many researchers have studied the lethal and sublethal effects of these and other insecticides on honeybees, and some of these studies have demonstrated a correlation between the insecticides and colony collapse disorder in bees. The authors investigated the effects of fipronil and imidacloprid on the bioenergetic functioning of mitochondria isolated from the heads and thoraces of Africanized honeybees. Fipronil caused dose-dependent inhibition of adenosine 5'-diphosphate-stimulated (state 3) respiration in mitochondria energized by either pyruvate or succinate, albeit with different potentials, in thoracic mitochondria; inhibition was strongest when respiring with complex I substrate. Fipronil affected adenosine 5'-triphosphate (ATP) production in a dose-dependent manner in both tissues and substrates, though with different sensitivities. Imidacloprid also affected state-3 respiration in both the thorax and head, being more potent in head pyruvate-energized mitochondria; it also inhibited ATP production. Fipronil and imidacloprid had no effect on mitochondrial state-4 respiration. The authors concluded that fipronil and imidacloprid are inhibitors of mitochondrial bioenergetics, resulting in depleted ATP. This action can explain the toxicity of these compounds to honeybees.

Enriquecimento da dieta do bicho-da-seda com extrato hidrossolúvel de soja / Enrichment of the silkworms diet with soybean water soluble extract

O objetivo deste trabalho foi avaliar se a suplementação da dieta do bicho-da-seda com extrato hidrossolúvel de soja (EHS) interfere na produção de casulos e no consumo de folhas de amoreira (Morus alba L.) de diferentes cultivares pelas lagartas. O delineamento experimental utilizado foi o inteiramente casualizado em esquema fatorial 4x5, com quatro cultivares de amoreira ('IZ64', 'IZ56/4', 'FM86' e 'FMSM') e cinco dietas (folhas não tratadas, folhas umedecidas com água, folhas enriquecidas com EHS, em três níveis: suplementação da dieta com 5, 10 e 20% de PB). O EHS foi aspergido sobre as folhas, tendo a água como agente dispersante. O consumo foi verificado pela diferença entre a quantidade de folhas fornecidas e a quantidade não consumida pelas lagartas. A ingestão de alimento apresentou-se de forma quadrática, diminuindo à medida que o EHS foi adicionado à dieta. Houve interação dos fatores cultivar e dieta. O fornecimento de folhas de amoreira com acréscimo proteico de 10% por meio do EHS melhorou a qualidade dos casulos do bicho-da-seda com a utilização das cultivares 'IZ56/4' e 'FMSM'. A pulverização de EHS nas folhas da cultivar 'IZ64' prejudicou a produção de casulos. Lagartas do bicho-da-seda, alimentadas com folhas de amoreira sem suplementação, produziram cascas séricas mais pesadas com folhas das cultivares 'IZ64' e 'FMSM'. A aspersão de água nas folhas incrementou o peso de cascas séricas produzidas a partir da cultivar 'FM86'. A suplementação da dieta com EHS diminui o consumo da lagarta do bicho-da-seda. A aplicação de EHS influencia a produção de casulos em função da cultivar utilizada.

The aim of this research was to assess if the supplementation of the silkworm diet with soybean water soluble extract (WSE) interferes on the consumption of mulberry leaves (Morus alba L.) by the caterpillars and on cocoon production. The experimental design was completely randomized in factorial arrangement 4x5, with four mulberry varieties ('IZ64', 'IZ56/4', and 'FMSM' 'FM86') and five diets (untreated control leaves, leaves moistened with water and leaves enriched with WSE, in three levels). The WSE was sprayed on the leaves with water as dispersing agent. The consumption was measured by the difference between the quantity of food provided and the amount that was not consumed by the larvae. The consumption was presented quadratic form, which decreased as the WSE was added on diet. There was interaction between cultivar and diets. Silkworm fed diets containing leaves of mulberry with addition of 10% of crude protein through WSE improved the quality of cocoons with utilization of 'IZ56/4' and 'FMSM' cultivars. The sprayed of WSE on leaves of 'IZ64' cultivar decreased production of cocoons. Silkworm fed with leaves without supplementation produced heavier cocoons shells of the 'IZ64' and 'FMSM' cultivars. Water sprayed on leaves provided higher weight of cocoons shells from 'FM86' cultivar. The WSE supplementation in diets decreases consumption of silkworm. The WSE sprayed influences cocoon production depending on the cultivar.

Estratégia alimentar com ciclos de restrição e realimentação no desempenho produtivo de juvenis de tilápia do Nilo da linhagem GIFT / Feeding strategies with cycles of restriction and refeeding on growth performance of Nile tilapia juveniles, GIFT line

O objetivo deste estudo foi avaliar a ocorrência de resposta compensatória no desempenho produtivo de juvenis de tilápia do Nilo Oreochromis niloticus, linhagem GIFT, submetidos a diferentes estratégias alimentares. Foram utilizados 135 juvenis de tilápia, distribuídos em nove tanques de polietileno de 100L cada. As estratégias testadas foram: grupo controle (alimentado todo dia), grupo alimentado por cinco dias seguidos de dois dias de restrição de alimento (5A/2R) e grupo alimentado por quatro dias seguidos de três dias de restrição de alimento (4A/3R). Foram avaliados parâmetros físico-químicos da água e de desempenho produtivo. Os resultados foram submetidos à análise de variância, e as médias foram comparadas pelo teste Tukey, a 5 por cento de probabilidade. A qualidade de água, o fator de condição e a conversão alimentar não foram influenciados pela estratégia alimentar. O grupo alimentado com a estratégia 5A/2R apresentou peso final, ganho de peso e taxa de crescimento específico semelhantes ao grupo continuamente alimentado (7,8 e 9,2g; 6,4 e 7,8g e 2,7 e 3,0 por cento dia-1, para peso final, ganho de peso e taxa de crescimento especifico, respectivamente). A estratégia 4A/3R apresentou os piores resultados de desempenho produtivo, e a estratégia 5A/2R pode ser usada na alimentação de juvenis de tilápia do Nilo sem prejuízo ao desempenho produtivo, possibilitando inclusive redução de até 22,5 por cento na quantidade de alimento ofertada.

The objective of this study was to evaluate the occurrence of compensatory answer on productive performance of Nile tilapia Oreochromis niloticus juveniles, GIFT line, submitted to different feeding strategies. It was used 135 juveniles tilapias distributed in nine 100L polyethylene tanks each. The tested strategies were the control group (fed everyday), group fed for 5 days followed by 2 days of food restriction (5A/2R) and group fed for 4 days followed by 3 days of food restriction (4A/3R). Were evaluated physical chemical water parameters e growth performance. Results were submitted to an analysis of variance and averages compared by Tukey test 5 percent. Water quality, condition factor and feed conversion were not influenced by feeding strategy. Group fed 5A/2R strategy showed final weight, weight gain and specific growth rate similar to group continually fed (7.8 e 9.2g; 6.4 e 7.8g e 2.7 e 3.0 percent day-1, for final weight, weight gain and specific growth rate, respectively). The strategy 4A/3R showed the poorest productive performance. Strategy 5A/2R can be used in Nile tilapia juveniles feeding without damage to the fish productive performance, allowing a reduction of up to 22.5 percent on the feed quantity offered.