Alterations in the Microbiota of Caged Honeybees in the Presence of Nosema ceranae Infection and Related Changes in Functionality

Several studies have outlined that changes in the honeybee gut microbial composition may impair important metabolic functions supporting the honeybees’ life. Gut dysbiosis may be caused by diseases like Nosema ceranae or by other anthropic, environmental or experimental stressors. The present work contributes to increasing knowledge on the dynamics of the gut microbiome acquisition in caged honeybees, an experimental condition frequently adopted by researchers, with or without infection with N. ceranae, and fed with a bacterial mixture to control N. ceranae development. Changes of the gut microbiota were elucidated comparing microbial profile of caged and open-field reared honeybees. The absolute abundance of the major gut microbial taxa was studied with both NGS and qPCR approaches, whereas changes in the functionality were based on RAST annotations and manually curated. In general, all caged honeybees showed important changes in the gut microbiota, with γ-proteobacteria (Frischella, Gilliamella and Snodgrassella) lacking in all caged experimental groups. Caged honeybees infected with N. ceranae showed also a strong colonization of environmental taxa like Citrobacter, Cosenzaea and Morganella, as well as possibly pathogenic bacteria such as Serratia. The colonization of Serratia did not occur in presence of the bacterial mixture. The functionality prediction revealed that environmental bacteria or the supplemented bacterial mixture increased the metabolic potential of the honeybee gut microbiome compared to field and caged controls

Honeybees exposure to natural feed additives: How is the gut microbiota affected?

The role of a balanced gut microbiota to maintain health and prevent diseases is largely established in humans and livestock. Conversely, in honeybees, studies on gut microbiota perturbations by external factors have started only recently. Natural methods alternative to chemical products to preserve honeybee health have been proposed, but their effect on the gut microbiota has not been examined in detail. This study aims to investigate the effect of the administration of a bacterial mixture of bifidobacteria and Lactobacillaceae and a commercial product HiveAlive™ on honeybee gut microbiota. The study was developed in 18 hives of about 2500 bees, with six replicates for each experimental condition for a total of three experimental groups. The absolute abundance of main microbial taxa was studied using qPCR and NGS. The results showed that the majority of the administered strains were detected in the gut. On the whole, great perturbations upon the administration of the bacterial mixture and the plant-based commercial product were not observed in the gut microbiota. Significant variations with respect to the untreated control were only observed for Snodgrassella sp. for the bacterial mixture, Bartonella sp. in HiveAlive™ and Bombilactobacillus sp. for both. Therefore, the studied approaches are respectful of the honeybee microbiota composition, conceivably without compromising the bee nutritional, social and ecological functions

Screening of dietary ingredients against the honey bee parasite Nosema ceranae

Nosema ceranae is a major pathogen in the beekeeping sector, responsible for nosemosis. This disease is hard to manage since its symptomatology is masked until a strong collapse of the colony population occurs. Conversely, no medicaments are available in the market to counteract nosemosis, and only a few feed additives, with claimed antifungal action, are available. New solutions are strongly required, especially based on natural methods alternative to veterinary drugs that might develop resistance or strongly pollute honey bees and the environment. This study aims at investigating the nosemosis antiparasitic potential of some plant extracts, microbial fermentation products, organic acids, food chain waste products, bacteriocins, and fungi. Honey bees were singularly infected with 5 × 104 freshly prepared N. ceranae spores, reared in cages and fed ad libitum with sugar syrup solution containing the active ingredient. N. ceranae in the gut of honey bees was estimated using qPCR. The results showed that some of the ingredients administered, such as acetic acid at high concentration, p-coumaric acid, and Saccharomyces sp. strain KIA1, were effective in the control of nosemosis. On the other hand, wine acetic acid strongly increased the N. ceranae amount. This study investigates the possibility of using compounds such as organic acids or biological agents including those at the base of the circular economy, i.e., wine waste production, in order to improve honeybee health

The fate of bacteria in urban wastewater-irrigated peach tree: a seasonal evaluation from soil to canopy

Irrigation with wastewater can be a solution to preserve and mitigate freshwater demand, in particular during drought periods. Unfortunately, wastewater, although being treated at different levels, could be a carrier of human pathogens (e.g., E. coli) and potentially contaminate crops for human consumptions.This study investigated the seasonal microbiological concentrations, on soil, shoot and fruit tissues of potted peach trees, following two irrigation treatments: freshwater (FW) and secondary urban wastewater without the final disinfection treatment (SW). E. coli was only detected in SW irrigated soil, whereas total coliforms (TC) and total bacteria counts (TBC) were similar in both treatments throughout the season. EndophyticE. coli, Salmonella spp. and TC were not detected in shoot and fruit, but a higher presence of total bacteria (TBC) was observed in SW-irrigated tree compared to FWirrigated tree. In particular, SW shoots had a higher load compared to fruits, thus showing a potential effect of leaf transpiration, that promoted the transfer of water-borne bacteria from soil to the epigeal part (shoot). The adoption of low-quality SW (even above the microbiological limits of the European Regulation 2020/741 for wastewater re-use in agriculture), when a drip irrigation method is applied, could be a valid alternative to save fresh water without compromising fruit safety

Study on a fermented whole wheat: Phenolic content, activity on PTP1B enzyme and in vitro prebiotic properties

Fermented cereals, staple foods in Asia and Africa, are recently receiving a growing interest in Western countries. The object of this work is the characterization of a fermented wheat used as a food ingredient and dietary supplement. To this aim, the phenolic composition, the activity on protein tyrosine phosphatase 1B (PTP1B), an enzyme overexpressed in type-II diabetes, the in vitro prebiotic properties on Lactobacillus reuteri and the microbial composition were investigated. Basic and acidic hydrolysis were tested for an exhaustive recovery of bound phenols: the acidic hydrolysis gave best yields. Methyl ferulate and neocarlinoside were identified for the first time in wheat. The inhibitory power of the extracts of several batches were investigated on PTP1B enzyme. The product was not able to inhibit the enzyme, otherwise, for the first time, a complete inhibition was observed for schaftoside, a major C-flavonoid of wheat. The microbial composition was assessed identifying Lactobacillus, Enterococcus, and Pediococcus as the main bacterial species. The fermented wheat was a suitable substrate for the grown of L. reuteri, recognized for its health properties in the human gut. The proposed method for phenols is easier compared to those based on strong basic hydrolysis; our results assessed the bound phenols as the major fraction, differently from that suggested by the literature for fermented cereals

Effects of a natural extract from Mangifera indica L, and its active compound, mangiferin, on energy state and lipid peroxidation of red blood cells

Following oxidative stress, modifications of several biologically important macromolecules have been demonstrated. In this study we investigated the effect of a natural extract from Mangifera indica L (Vimang), its main ingredient mangiferin and epigallocatechin gallate (EGCG) on energy metabolism, energy state and malondialdehyde (MDA) production in a red blood cell system. Analysis of NIDA, high energy phosphates and ascorbate was carried out by high performance liquid chromatography (HPLC). Under the experimental conditions, concentrations of NIDA and ATP catabolites were affected in a dose-dependent way by H2O2. Incubation with Vimang (0.1, 1, 10, 50 and 100 mu g/mL), mangiferin (1, 10, 100 mu g/mL) and EGCG (0.01, 0.1, 1, 10 mu M) significantly enhances erythrocyte resistance to H2O2-induced reactive oxygen species production. In particular, we demonstrate the protective activity of these compounds on ATP, GTP and total nucleotides (NT) depletion after H2O2-induced damage and a reduction of NAD and ADP, which both increase because of the energy consumption following H2O2 addition. Energy charge potential, decreased in H2O2-treated erythrocytes, was also restored in a dose-dependent way by these substances. Their protective effects might be related to the strong free radical scavenging ability described for polyphenols. (c) 2006 Elsevier B.V All rights reserved

Simple system using natural mineral water for high-throughput phenotyping of Arabidopsis thaliana seedlings in liquid culture

Background: Phenotyping for plant stress tolerance is an essential component of many research projects. Because screening of high numbers of plants and multiple conditions remains technically challenging and costly, there is a need for simple methods to carry out large-scale phenotyping in the laboratory.Methods: We developed a method for phenotyping the germination and seedling growth of Arabidopsis (Arabidopsis thaliana) Col-0 in liquid culture. Culture was performed under rotary shaking in multiwell plates, using Evian natural mineral water as a medium. Nondestructive and accurate quantification of green pixels by digital image analysis allowed monitoring of growth. Results: The composition of the water prevented excessive root elongation growth that would otherwise lead to clumping of seedlings observed when classic nutrient-rich medium or deionized water is used. There was no need to maintain the cultures under aseptic conditions, and seedlings, which are photosynthetic, remained healthy for several weeks. Several proof-of-concept experiments demonstrated the usefulness of the approach for environmental stress phenotyping. Conclusion: The system described here is easy to set up, cost-effective, and enables a single researcher to screen large numbers of lines under various conditions. The simplicity of the method clearly makes it amenable to high-throughput phenotyping using robotics