Long-term stability and bactericidal properties of galenic formulations of Cannabis sativa oils.

The long-term stability in real and accelerated time for galenic oils based on full-spectrum cannabis has been studied, using sesame oil as a dilutant. Sesame oil is one of the most used vehicles in the cannabis pharmaceutical industry due to the costs and increased oral bioavailability of cannabinoids. The real-time assays conducted at 25 °C over twelve months demonstrated high stability and showed no significant changes in the composition of cannabinoids, total polyphenols, flavonoids, or antioxidant capacity. In these studies, it was observed that there was no development of microorganisms compromising the stability of the oils over a year. The three oil varieties exhibited a high bactericidal capacity against E. coli, S. aureus, and P. larvae.

Evaluation of the Protoscolicidal Activity of Humulus lupulus Methanolic Extracts on Echinococcus granulosus Sensu Stricto.

The larval stage of the parasite Echinococcus granulosus sensu lato (s.l) is responsible for cystic echinococcosis (CE), a long-term infection affecting humans and animals worldwide, and constitutes a serious public health concern. If left untreated, CE can cause serious damage to multiple organs, especially the liver and lungs. Regarding the treatment, in the last few years, the use of pharmacological treatment has increased, suggesting that in the future, drug therapy may replace surgery for uncomplicated cysts. However, the only available anthelmintic drug to treat this infection is the albendazole, which has an efficacy that does not exceed 50%. On the basis of the above-mentioned evidence, new and improved alternative treatments are urgently needed. The use of natural products and their active fractions and components holds great promise as a valuable resource for the development of novel and effective therapies. Hop (Humulus lupulus L.) is a bittering agent in the brewing industry for which the sedative, digestive, anti-inflammatory, and antimicrobial effects have been reported. The purpose of this study was to assess the in vitro efficacy of methanolic extracts from the leaves of hop varieties against E. granulosus sensu stricto (s.s) protoscoleces. Varieties Mapuche and Victoria caused a stronger protoscolicidal effect compared to the Bullion, Cascade, and Traful varieties (P < 0.01), coinciding with their highest content of flavonoids, total polyphenols, and saponins. The viability of protoscoleces treated with the varieties Mapuche and Victoria decreased to approximately 50% at days 5 y 8, respectively, showing alterations such as soma contraction and impaired microtriches. After 18 days of treatment with both varieties, protoscoleces were completely altered both structurally and ultrastructurally. In conclusion, the methanolic extracts of the H. lupulus varieties Mapuche and Victoria demonstrated a marked in vitro effect against E. granulosus s.s. protoscoleces. The beer-making industry exclusively uses hop cones, leaving behind large amounts of hop leaves as an agricultural by-product that is not being utilized. On the basis of our study, we propose that hop leaves could also be used as a source of secondary metabolites with anthelmintic activity.

Anthelmintic Effect of Cannabidiol against Echinococcus granulosus sensu stricto.

Cystic echinococcosis is a global parasitic zoonosis caused by infection with the larval stage of Echinococcus granulosus sensu lato. Cystic echinococcosis affects more than 1 million people worldwide, causing important economic costs in terms of management and livestock associated losses. Albendazole is the main drug used in treating human cystic echinococcosis. In spite of this, its low aqueous solubility, poor absorption, and consequently erratic bioavailability are the cause of its chemotherapeutic failures. Based on the described problem, new treatment alternatives urgently need to be developed. The aim of the present research was to study the in vitro and in vivo efficacy of cannabidiol (CBD), the second most abundant component of the Cannabis sativa plant, was demonstrated against E. granulosus sensu stricto. CBD (50 µg/mL) caused a decrease in protoscoleces viability of 80 % after 24 h of treatment which was consistent with the observed tegumental alterations. Detachment of the germinal layer was observed in 50 ± 10% of cysts treated with 50 µg/mL of CBD during 24 h. In the clinical efficacy study, all treatments reduced the weight of cysts recovered from mice compared with the control group. However, this reduction was only significant with ABZ suspension and the CBD + ABZ combination. As we could observe by the SEM study, the co-administration of CBD with ABZ suspension caused greater ultrastructural alteration of the germinal layer in comparison with that provoked with the monotherapy. Further in vivo research will be conducted by changing the dose and frequency of CBD and CBD + ABZ treatments and new available CBD delivery systems will also be assayed to improve bioavailability in vivo.

Broad-Spectrum Antiviral Effect of Cannabidiol Against Enveloped and Nonenveloped Viruses.

Introduction: Cannabidiol (CBD), the main non-psychoactive cannabinoid of the Cannabis sativa plant, is a powerful antioxidant compound that in recent years has increased interest due to causes effects in a wide range of biological functions. Zika virus (ZIKV) is a virus transmitted mainly by the Aedes aegypti mosquitoes, which causes neurological diseases, such as microcephaly and Guillain-Barre syndrome. Although the frequency of viral outbreaks has increased recently, no vaccinations or particular chemotherapeutic treatments are available for ZIKV infection. Objectives: The major aim of this study was to explore the in vitro antiviral activity of CBD against ZIKV, expanding also to other dissimilar viruses. Materials and Methods: Cell cultures were infected with enveloped and nonenveloped viruses and treated with non-cytotoxic concentrations of CBD and then, viral titers were determined. Additionally, the mechanism of action of the compound during ZIKV in vitro infections was studied. To study the possible immunomodulatory role of CBD, infected and uninfected Huh-7 cells were exposed to 10 µM CBD during 48 h and levels of interleukins 6 and 8 and interferon-beta (IFN-ß) expression levels were measured. On the other hand, the effect of CBD on cellular membranes was studied. For this, an immunofluorescence assay was performed, in which cell membranes were labeled with wheat germ agglutinin. Finally, intracellular cholesterol levels were measured. Results: CBD exhibited a potent antiviral activity against all the tested viruses in different cell lines with half maximal effective concentration values (CE50) ranging from 0.87 to 8.55 µM. Regarding the immunomodulatory effect of CBD during ZIKV in vitro infections, CBD-treated cells exhibited significantly IFN-ß increased levels, meanwhile, interleukins 6 and 8 were not induced. Furthermore, it was determined that CBD affects cellular membranes due to the higher fluorescence intensity that was observed in CBD-treated cells and lowers intracellular cholesterol levels, thus affecting the multiplication of ZIKV and other viruses. Conclusions: It was demonstrated that CBD inhibits structurally dissimilar viruses, suggesting that this phytochemical has broad-spectrum antiviral effect, representing a valuable alternative in emergency situations during viral outbreaks, like the one caused by severe acute respiratory syndrome coronavirus 2 in 2020.

Hydrolats from Humulus lupulus and Their Potential Activity as an Organic Control for Varroa destructor.

Varroa destructor is a parasitic mite, which is considered a severe pest for honey bees causing serious losses to beekeeping. Residual hydrolats from steam extraction of hop essential oils, generally considered as a waste product, were tested for their potential use as acaricides on V. destructor. Four hop varieties, namely Cascade, Spalt, Victoria, and Mapuche, showed an interesting performance as feasible products to be used in the beekeeping industry. Some volatile oxidized terpenoids were found in the hydrolats, mainly ß-caryophyllene oxide, ß-linalool, and isogeraniol. These compounds, together with the presence of polyphenols, flavonoids, and saponins, were probably responsible for the promissory LC50 values obtained for mites after hydrolat exposition. Victoria hydrolat was the most toxic for mites (LC50: 16.1 µL/mL), followed by Mapuche (LC50 value equal to 30.1 µL/mL), Spalt (LC50 value equal to 114.3 µL/mL), and finally Cascade (LC50: 117.9 µL/mL). Likewise, Spalt had the highest larval survival, followed by Victoria and Mapuche. Cascade was the variety with the highest larval mortality. In addition, none of the extracts showed mortality higher than 20% in adult bees. The Victoria hydrolat presented the best results, which makes it a good compound with the prospect of an acaricide treatment against V. destructor.

Behavioural factors underlying innovative problem-solving differences in an avian predator from two contrasting habitats.

Innovative behavior is considered one of the main factors facilitating the adaptation of animals to urban life. However, the relationship between urbanization and innovativeness is equivocal, perhaps reflecting aspects of urban environments that influence differently the behavioural traits underlying the occurrence of an innovation. In this work, we analysed the variation in innovative problem-solving performance between urban and rural individuals of the Caracara Chimango (Milvago chimango), with the goal of determining which behavioural trait (or combination) most explained such variation. We found that urban raptors outperformed rural ones in their solving speed and solving level (number of solutions) with a multiaccess box. They also showed more persistence, motor flexibility and diversity, as well as higher effectiveness in their solving attempts than rural chimangos. Sex was not an important factor. Urban chimangos showed less neophobia and spent more time exploring the box than rural birds during the initial habituation period, which probably determined the amount of information about the system that each individual had at the beginning of first problem solving trial. This difference in novelty response both directly and indirectly, through its relationship with persistence, motor flexibility and proportion of effective attempts, explained variability in solving performance. All individuals showed a decrease in solving latency, and an increase in solving level with experience, indicating that learning occurred in both raptor groups. This improvement occurred in parallel with changes in the afore-mentioned traits, though the pattern of improvement differed between urban and rural chimangos. We suggest that the characteristics of urban areas modulate the novelty response of chimangos, along with other correlated non-cognitive behavioural traits, which act in combination to increase the chances that novel problems could be quickly solved, and the resulting new behaviours established in city populations of this species.