Spent coffee grounds extract: antimicrobial activity against Paenibacillus larvae and its effect on the expression of antimicrobial peptides in Apis mellifera.

In recent years, natural alternatives have been sought for the control of beekeeping pathologies; in the case of American Foulbrood (AFB) disease, the use of synthetic antibiotics was prohibited due to honey contamination and the generation of resistant bacteria. The significant increase in population growth worldwide has led to great concern about the production of large amounts of waste, including those from agribusiness. Among the most important beverages consumed is coffee, generating thousands of tons of waste called spent coffee grounds (SCG). The SCG is a source of many bioactive compounds with known antimicrobial activity. The aims of the present work were: (1) to obtain and chemically analyse by HPLC of SCG extracts (SCGE), (2) to analyse the antimicrobial activity of SCGE against vegetative form of Paenibacillus larvae (the causal agent of AFB), (3) to evaluate the toxicity in bees of SCGE and (4) to analyse the effect of the extracts on the expression of various genes of the immune system of bees. SCGs have a high content of phenolic compounds, and the caffeine concentration was of 0.3%. The MIC value obtained was 166.667 µg/mL; the extract was not toxic to bees, and interestingly, overexpression of abaecin and hymenoptaecin peptides was observed. Thus, SCGE represents a promising alternative for application in the control of American Foulbrood and as a possible dietary supplement to strengthen the immune system of honeybees. Therefore, the concept of circular bio-economy could be applied from the coffee industry to the beekeeping industry.

Antifungal activity of "HO21-F", a formulation based on Olea europaea plant extract, in honey bees infected with Nosema ceranae.

Nosema ceranae is a microsporidium parasite that silently affects honey bees, causing a disease called nosemosis. This parasite produces resistant spores and germinates in the midgut of honey bees, extrudes a polar tubule that injects an infective sporoplasm in the host cell epithelium, proliferates, and produces intestinal disorders that shorten honey bee lifespan. The rapid extension of this disease has been reported to be widespread among adult bees, and treatments are less effective and counterproductive weakening colonies. This work aimed to evaluate the antifungal activity of a prototype formulation based on a non-toxic plant extract (HO21-F) against N. ceranae. In laboratory, honey bees were infected artificially, kept in cages for 17 days and samples were taken at 7 and 14 days post infection (dpi). At the same time, in field conditions we evaluated the therapeutic effect of HO21-F for 28 days in naturally infected colonies. The effectiveness of the treatment has been demonstrated by a reduction of 83.6 % of the infection levels observed in laboratory conditions at concentrations of 0.5 and 1 g/L without affecting the survival rate. Besides, in-field conditions we reported a reduction of 88 % of the infection level at a concentration of 2.5 g/L, obtaining better antifungal effectiveness in comparison to other commercially available treatments. As a result, we observed that the use of HO21-F led to an increase in population size and honey production, both parameters associated with colony strength. The reported antifungal activity of HO21-F against N. ceranae, with a significant control of spore proliferation in worker bees, suggests the promising commercial application use of this product against nosemosis, and it will encourage new research studies to understand the mechanism of action, whether related to the spore-inhibition effect and/or a stimulating effect in natural response of colonies to counteract the disease.

Protein Serine/Threonine Phosphatase Type 2C of Leishmania mexicana.

Protein phosphorylation and dephosphorylation are increasingly recognized as important processes for regulating multiple physiological mechanisms. Phosphorylation is carried out by protein kinases and dephosphorylation by protein phosphatases. Phosphoprotein phosphatases (PPPs), one of three families of protein serine/threonine phosphatases, have great structural diversity and are involved in regulating many cell functions. PP2C, a type of PPP, is found in Leishmania, a dimorphic protozoan parasite and the causal agent of leishmaniasis. The aim of this study was to clone, purify, biochemically characterize and quantify the expression of PP2C in Leishmania mexicana (LmxPP2C). Recombinant LmxPP2C dephosphorylated a specific threonine (with optimal activity at pH 8) in the presence of the manganese divalent cation (Mn+2). LmxPP2C activity was inhibited by sanguinarine (a specific inhibitor) but was unaffected by protein tyrosine phosphatase inhibitors. Western blot analysis indicated that anti-LmxPP2C antibodies recognized a molecule of 45.2 kDa. Transmission electron microscopy with immunodetection localized LmxPP2C in the flagellar pocket and flagellum of promastigotes but showed poor staining in amastigotes. Interestingly, LmxPP2C belongs to the ortholog group OG6_142542, which contains only protozoa of the family Trypanosomatidae. This suggests a specific function of the enzyme in the flagellar pocket of these microorganisms.