Tracing the origin and authenticity of Spanish PDO honey using metagenomics and machine learning.

The Protected Designation of Origin (PDO) indication for foods intends to guarantee the conditions of production and the geographical origin of regional products within the European Union. Honey products are widely consumed due to their health-promoting properties and there is a general interest in tracing their authenticity. In this regard, metagenomics sequencing and machine learning (ML) have been proposed as complementary technologies to improve the traceability methods of foods. Therefore, the aim of this study was to analyze the metagenomic profiles of Spanish honeys from three different PDOs (Granada, Tenerife and Villuercas-Ibores), and compare them with non-PDO honeys using ML models (PLS, RF, LOGITBOOST, and NNET). According to the results obtained, non-PDO honeys and Granada PDO showed higher beta diversity values than Tenerife and Villuercas-Ibores PDOs. ML classification of honey products allowed the identification of different microbial biomarkers of the geographical origin of honeys: Lactobacillus kunkeei, Parasaccharibacter apium and Lactobacillus helsingborgensis for PDO honeys and Paenibacillus larvae, Lactobacillus apinorum and Klebsiella pneumoniae for non-PDO honeys. In addition, potential microbial biomarkers of some honey varieties including L. kunkeei for Albaida and Retama del Teide varieties, and P. apium for Tajinaste variety, were identified. ML models were validated on an independent set of samples leading to high accuracy rates (above 90 %). This work demonstrates the potential of ML to differentiate different types of honey using metagenome-based methods, leading to high performance metrics. In addition, ML models discriminate both the geographical origin and variety of products corresponding to different PDOs and non-PDO products. Results here presented may contribute to develop enhanced traceability and authenticity methods that could be applied to a wide range of foods.

Effects of ground transport on the presence of heavy metals in selected honeybee products.

Honeybees are insects very sensitive to environmental pollution and at the same time very good indicators of the pollution levels for certain types of pollutants. The morphology and ethology of these insects make them perfect vectors for dust and substances, including heavy metals produced by anthropic activities or naturally generated and deposited on foraged flora. When bees are raised to produce foods such as honey and pollen, they can easily transfer pollutants collected from contaminated flower affecting the quality of these products. However, depending on geographical location of the apiaries and their distance from pollution sources, the risk to contaminate bee products can be higher or lower requiring deep investigations. In this study, two apiaries were built near ground transport infrastructures and used as monitoring stations for investigating heavy metal presence in beehive products such as bee wax, pollen, and honey. Another apiary was placed between these two locations at a distance of 500 m from each one and used as central node to asses possible diffusion trends. Parallel, air quality was monitored in the proximity of each apiary to verify the air pollution of the environments close to these sites. The results of the study suggest that the presence of the highway and the train station affected the levels of heavy metal presence in the apiary products. Air quality near apiaries was also negatively affected by ground transport, especially in proximity of the highway. Wax resulted significantly more polluted in the apiary close to train station with elements such as Al, Zn, and Ni, while honey and pollen were significantly more polluted in the proximity of the highway with elements such as Al, Fe, Cu, and Zn. Honey was the product suffering less the contamination by heavy metals while pollen was the worse. In conclusion, the presence of transportation nodes determined a higher accumulation of heavy metals in beehive products respect the apiary placed in between, suggesting to pay particular attention in the site selection for the placement of apiaries to protect both bees and human health.

A fluorescent NBD "turn-on" probe for the rapid and on-site analysis of fructose in food.

High fructose intake is an important cause of metabolic disease. Due to the increasing prevalence of metabolic diseases worldwide, the development of an accurate and efficient tool for monitoring fructose in food is urgently needed to control the intake of fructose. Herein, a new fluorescent probe NBD-PQ-B with 7-nitrobenz-2-oxa-1, 3-diazole (NBD) as the fluorophore, piperazine (PQ) as the bridging group and phenylboronic acid (B) as the recognition receptor, was synthesized to detect fructose. The fluorescence of NBD-PQ-B increased linearly at 550 nm at an excitation wavelength of 497 nm with increasing fructose concentration from 0.1 to 20 mM. The limit of detection (LOD) of fructose was 40 µM. The pKa values of NBD-PQ-B and its fructose complexes were 4.1 and 10.0, respectively. In addition, NBD-PQ-B bound to fructose in a few seconds. The present technique was applied to determine the fructose content in beverages, honey, and watermelon with satisfactory results. Finally, the system could not only be applied in an aqueous solution with a spectrophotometer, but also be fabricated as a NBD-PQ-B/polyvinyl oxide (PEO) film by electrospinning for on-site food analysis simply with the assistance of a smartphone.

The first ITS1 profiling of honey samples from the Southeast Asian region Lombok, Bali and Banggi Island.

Southern Asian flowers offer honeybees a diversity of nectar. Based on its geographical origin, honey quality varies. Traditional methods are less authentic than DNA-based identification. The origin of honey is determined by pollen, polyphenolic, and macro-microorganisms. In this study, amplicon sequencing targets macro-microorganisms in eDNA using the ITS1 region to explore honey's geographical location and authentication. The variety of honey samples was investigated using ITS1 with Illumina sequencing. For all four honey samples, raw sequence reads showed 979,380 raw ITS1 amplicon reads and 375 ASVs up to the phylum level. The highest total number of 202 ASVs up to phylum level identified Bali honey with 211,189 reads, followed by Banggi honey with 309,207 a total number of 111 ASVs, and Lombok represents only 63 ASVs up to phylum level with several read 458,984. Based on Shannon and Chao1, honey samples from Bali (B2) and (B3) exhibited higher diversity than honey from Lombok (B1) and green honey from Sabah (B4), while the Simpson index showed that Banggi honey (B4) had higher diversity. Honey samples had significant variance in mycobiome taxonomic composition and abundance. Zygosaccharomyces and Aspergillus were the main genera found in Lombok honey, with percentages of 68.81% and 29.76% respectively. Bali honey samples (B2 and B3) were identified as having a significant amount of the genus Aureobasidium, accounting for 40.81% and 25% of the readings, respectively. The microbiome composition of Banggi honey (B4) showed a high presence of Zygosaccharomyces 45.17% and Aureobasidium 35.24%. The ITS1 analysis effectively distinguishes between honey samples of different origins and its potential as a discriminatory tool for honey origin and authentication purposes.

Pollen analysis of honey and pollen stored by Melipona (Melikerria) fasciculata Smith, 1854 (Apidae Meliponini), in an Amazon and Cerrado transition area, Maranhão, Brazil.

The identification of meliponicultural flora is fundamental for the preservation of bees, as well as enabling the development of pollinator management and reforestation programs. In this context, this study aimed to investigate the types of pollen found in the honey and pollen pots of the Melipona fasciculata bee in Arari, State of Maranhão, between August 2022 and July 2023. 40 types of pollen were identified, distributed among 18 botanical families, with Fabaceae standing out (46.15% in pollen pots and 37.86% in honey pots), Myrtaceae (11.53% in pollen and 13.51% in honey) and Rubiaceae (7.69% in pollen and 5.40% in honey). Sapindaceae, in the honey pots, accounted for 5.40%. The other families showed lower percentages, 3.84% in the pollen pots and 2.70% in the honey pots. In terms of similarity, two distinct groups were observed in both the pollen and honey pots. In the pollen pots, group A (May-June ~ 97%) and group B (Feb-Mar ~ 99%) stood out, while in the honey pots, group A (Mar-Apr ~ 98%) and group B (Jun-Jul-Sept ~ 98%) showed the highest percentages. These findings highlight the wide range of resources used by the M. fasciculata species, as well asits preference for Fabaceae and Myrtaceae, due to the diversity and availability of trophic resources. An understanding of the meliponicultural flora is essential to support effective conservation strategies, which aim not only to guarantee the survival of the bees, but also to ensure the continued production of honey, a resource of great importance to local communities.

Antibacterial Effect of Spanish Honeys of Different Botanical Origins against Staphylococcus epidermidis.

Honey is traditionally used for its medicinal properties attributed to its antibacterial and antioxidant effects. It is considered a natural alternative to conventional antibiotics. This effect has been attributed to their physico-chemical properties, as various chemical parameters can synergistically influence this effect. The aim of this study is to assess Spanish honeys of diverse botanical origins for their antibacterial efficacy against Staphylococcus epidermidis, correlating their physico-chemical attributes, (poly)phenol content, and antioxidant activity. The methods included colour determination via two methodologies, acidity, pH, moisture content, and sugar concentration. (Poly)phenol content was quantified using the Folin-Ciocalteau method, while antioxidant activity was evaluated via the FRAP method. Subsequently, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against S. epidermidis were investigated with different concentrations of honeys. The results revealed a direct relationship between honey darkness, (poly)phenol concentration, antioxidant activity, and antibacterial efficacy. Darker honeys exhibited higher (poly)phenol levels, greater antioxidant activity, and consequently, lower MIC and MBC values, showing enhanced antibacterial properties. These findings underscore the potential of honey as a therapeutic agent against S. epidermidis, particularly in wound healing applications to avoid infection. Further research into honey's multifaceted properties is warranted to unveil novel therapeutic avenues in healthcare.

Improved stability and specificity of baits for oral administration of substances to wild boar.

Oral vaccination is one of the most effective interventions for disease control in wildlife. As a result of the recent global reemergence of African swine fever and ongoing classical swine fever and animal tuberculosis, oral vaccination of Eurasian wild boar (Sus scrofa) receives increased interest. Several baits for wild boar and feral pigs have been described, but developing more stable and personalized formulations is important. This paper proposes a new bait formulation primarily composed of corn flour, piglet feed, sugar, and honey as a binder to obtain improved elasticity. The bait consists of a matrix with no protective coats, has a hemispherical shape (ø 3.4 ×1.6 cm), and displays an anise aroma and blue color. The color and aroma did not affect bait choice by wild boar, while bait coloring contributed to avoid consumption by non-target species (corvids). Baits with the new formulation were significantly more resistant to humidity and high temperatures than previous versions. Simulations suggest that baits with the new formulation are elastic enough to resist impacts from a maximum altitude of 750 m. Thus, the new bait prototype solves several problems of previous bait formulations while keeping a format that can be selectively consumed by piglets and adult wild boar.

Following Regulation, Imidacloprid Persists and Flupyradifurone Increases in Nontarget Wildlife.

After regulation of pesticides, determination of their persistence in the environment is an important indicator of effectiveness of these measures. We quantified concentrations of two types of systemic insecticides, neonicotinoids (imidacloprid, acetamiprid, clothianidin, thiacloprid, and thiamethoxam) and butenolides (flupyradifurone), in off-crop nontarget media of hummingbird cloacal fluid, honey bee (Apis mellifera) nectar and honey, and wildflowers before and after regulation of imidacloprid on highbush blueberries in Canada in April 2021. We found that mean total pesticide load increased in hummingbird cloacal fluid, nectar, and flower samples following imidacloprid regulation. On average, we did not find evidence of a decrease in imidacloprid concentrations after regulation. However, there were some decreases, some increases, and other cases with no changes in imidacloprid levels depending on the specific media, time point of sampling, and site type. At the same time, we found an overall increase in flupyradifurone, acetamiprid, thiamethoxam, and thiacloprid but no change in clothianidin concentrations. In particular, flupyradifurone concentrations observed in biota sampled near agricultural areas increased twofold in honey bee nectar, sevenfold in hummingbird cloacal fluid, and eightfold in flowers after the 2021 imidacloprid regulation. The highest residue detected was flupyradifurone at 665 ng/mL (parts per billion [ppb]) in honey bee nectar. Mean total pesticide loads were highest in honey samples (84 ± 10 ppb), followed by nectar (56 ± 7 ppb), then hummingbird cloacal fluid (1.8 ± 0.5 ppb), and least, flowers (0.51 ± 0.06 ppb). Our results highlight that limited regulation of imidacloprid does not immediately reduce residue concentrations, while other systemic insecticides, possibly replacement compounds, concurrently increase in wildlife. Environ Toxicol Chem 2024;43:1497-1508. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Physicochemical Characterization, Antioxidant and Tyrosinase Inhibitory Activities of Coffea Robusta Monofloral Honey from Dak Lak Province, Vietnam.

Robusta coffee blossom honey stands as a key regional product in Dak Lak province, Vietnam. Despite its significance, there exists a dearth of scientific data for assessing its quality. This study aims to fill this gap by characterizing the physicochemical properties and biological activities of coffee blossom honeys from three distinct sub-regions within Dak Lak province, Vietnam. These activities include ferric reducing power (FRP), DPPH and ABTS radical scavenging, as well as tyrosinase inhibitory activities. Moreover, the study compares these honey samples with other popular varieties in Vietnam, such as Lychee and Longan honeys. The physicochemical parameters of the honey samples meet the standards set by Codex Alimentarius 2001. Through UPLC analysis, eleven compounds were identified, with caffeine serving as a marker for coffee honey. Furthermore, by employing multiple factor analysis (MFA), it was observed that certain physicochemical properties correlate positively with tyrosinase inhibitory, DPPH, ABTS free radicals scavenging activities, and FRP. Notably, tyrosinase inhibitory activity exhibited a positive correlation with antioxidant activity. These findings underscore the high quality of Coffea robusta honey, showcasing its potent antioxidant and tyrosinase inhibitory activities.

*The traditional Chinese medicine processing change chemical composition and pharmacological effectiveness: Taking Atractylodes macrocephala Koidz. and honey bran-fried Atractylodes macrocephala Koidz. as examples.

BACKGROUND: Atractylodes macrocephala Koidz. (Baizhu in Chinese, BZ) is a typical traditional edible-medicinal herb used for thousands of years. Known as "the spleen-reinforcing medicine", it is often used clinically to treat reduced digestive function, abdominal distension, and diarrhoea, which are all caused by spleen deficiency. Among BZ's processing products, honey bran-fried BZ (HBBZ) is the only processed product recorded in BZ in the 2020 Chinese Pharmacopoeia (ChP). There are differences in effectiveness, traditional application, and clinical indications between them. PURPOSE: This review reviewed BZ and its main product HBBZ from botany, ethnopharmacology, chemical composition, pharmacological effectiveness, and safety. The changes in chemical composition and pharmacological effectiveness of BZ induced by the processing of traditional Chinese medicine were emphatically described. METHODS: Keywords related to Atractylodes macrocephala Koidz., honey bran frying, essential oil, lactones, polysaccharide and combinations to include published studies of BZ and HBBZ from 2004-2023 were searched in the following databases: Pubmed, Chengdu University of TCM Library, Google Scholar, China National Knowledge Infrastructure (CNKI), and Wanfang database. All studies, published in English or Chinese, were included. However, in the process of chemical composition collection, we reviewed all available literature on the chemical composition of BZ and HBBZ. CONCLUSION: Honey bran frying processing methods will affect BZ's chemical composition and pharmacological effectiveness. The types and contents of chemical components in the HBBZ showed some changes compared with those in BZ. For example, the content of volatile oil decreased and the content of lactones increased after stir-fried bran. In addition, new ingredients such as phenylacetaldehyde, 2-acetyl pyrrole, 6- (1,1-dimethylethyl) -3,4-dihydro-1 (2H) -naphthalone and 5-hydroxymethylfurfural appeared. Both BZ and HBBZ have a variety of pharmacological effectiveness. After stir-fried with honey bran, the "Zao Xing" is reduced, and the efficacy of tonify spleen is strengthened, which is more suitable for patients with weak spleen and stomach.

Integrated assessment of the spatial distribution, sources, degradation, and human risk of tetracyclines in honey in China.

Tetracyclines are widely used in Chinese apiculture. However, limited information is available on the presence of tetracycline residues in honey and the sources, degradation patterns, and associated health risks of these compounds. In this study, the presence of tetracyclines in honey samples across China was investigated over a four-year period. Additionally, the risks of dietary intake, as well as the sources and degradation patterns of tetracyclines in honey, were assessed. The three-dimensional spatial distributions (floral region, geographical region and entomological origin) of tetracyclines contamination varied significantly. Tetracycline residues in honey posed a moderate risk to children aged 3-10 years in Northwest China. Source analysis indicated that colony migration serves as the primary source of tetracyclines in honey. Based on the degradation patterns of tetracyclines in honey within colonies and during storage, oxytetracycline is more readily degraded than other tetracyclines. The main degradation products of tetracyclines are epimers and dehydration products, and the effects of these products on human health and the environment should be further evaluated in future studies. This comprehensive investigation provides valuable insights into the safe use and regulation of tetracyclines in Chinese apiculture.

Silver nanopopcorns decorated on flexible membrane for SERS detection of nitrofurazone.

The synthesis of three-dimensional silver nanopopcorns (Ag NPCs) onto a flexible polycarbonate membrane (PCM) for the detection of nitrofurazone (NFZ) on the fish surface by surface-enhanced Raman spectroscopy (SERS) is presented. The proposed flexible Ag-NPCs/PCM SERS substrate exhibits significant Raman signal intensity enhancement with the measured enhancement factor of 2.36 × 106. This is primarily attributed to the hotspots created on Ag NPCs, including numerous nanoscale protrusions and internal crevices distributed across the surface of Ag NPCs. The detection of NFZ by this flexible SERS substrate demonstrates a low limit of detection (LOD) of 3.7 × 10-9 M and uniform and reproducible Raman signal intensities with a relative standard deviation below 8.34%. It also exhibits excellent stability, retaining 70% of its efficacy even after 10 days of storage. Notably, the practical detection of NFZ in tap water, honey water, and fish surfaces achieves LOD values of 1.35 × 10-8 M, 5.76 × 10-7 M, and 3.61 × 10-8 M, respectively,  which highlights its effectiveness across different sample types. The developed Ag-NPCs/PCM SERS substrate presents promising potential for sensitive SERS detection of toxic substances in real-world samples.

Potentially functional lactose-free ice cream with Lacticaseibacillus casei CSL3, ginger, and honey.

AIMS: To develop and characterize a functional lactose-free ice cream with added ginger and honey, evaluate the survival of Lacticaseibacillus casei CSL3 under frozen storage and the simulated gastrointestinal tract (GIT), as well as antioxidant activity and product acceptability. METHODS AND RESULTS: The survival of Lacticaseibacillus casei CSL3 was evaluated for 180 days, under frozen storage, and GIT at 60 days. At 15 days of storage, proximal composition, antioxidant activity, color, pH, acidity, fusion, density, overrun, and sensory analysis were performed. Ice cream was an effective food matrix for maintaining the viability of CSL3, with concentrations > 7 log CFU g- 1 during storage and GIT. In addition, the analysis showed overrun and prebiotic characteristics through high values of antioxidant activity and phenolic compounds, good acceptability, and purchase intention. CONCLUSIONS: The product has satisfactory market potential (acceptance rate of 95.19% and purchase intention rate > 96%), and it could become another means of inserting probiotics in food.

Dual-ligand lanthanide metal-organic framework based ratiometric fluorescent platform for visual monitoring of aminoglycoside residues in food samples.

Herein, a dual-emission Eu metal-organic framework (Eu-MOF) is prepared and used as the ratiometric fluorescence probe for ultrasensitive detection of aminoglycoside antibiotics (AGs). Due to the strong hydrogen bond interactions between AGs and Eu-MOF, the blue emission is enhanced while the red emission has little fluctuation in Eu-MOF with the addition of AGs, thus a good linear relationship with the logarithm of AGs concentrations from 0.001 to 100 µg/mL can be established for quantitative analysis. Good sensitivity with the detection limit of 0.33 ng/mL for apramycin, 0.32 ng/mL for amikacin and 0.30 ng/mL for kanamycin is achieved. The proposed assay demonstrates good selectivity and applicability for determination of AGs in real milk and honey samples. The Eu-MOF materials are further fabricated as fluorescent test papers for facile visual detection. The as-established ratio fluorescence platform offers a portable and economical way for rapid monitoring AGs residues in complex food samples.

Deep eutectic solvent-assisted synthesis of La-Ce hybrid nanorods for the colorimetric determination of tetracycline in foods.

Tetracycline (TC) as a broad-spectrum antibiotic, is widely used in the prevention and treatment of various bacterial diseases. However, its abuse in the livestock industry may lead to interference in human microecology, thereby causing various side effects. In this study, deep eutectic solvents (DESs) were synthesized using L-(-)-threonine (L-(-)-Thr) and cerium nitrate hexahydrate (Ce(NO3)3·6H2O), and later lanthanum nitrate hexahydrate (La(NO3)3·6H2O) was doped to synthesize La-Ce hybrid nanorods. These nanorods can be used for the determination of TC with high sensitivity and selectivity by the colorimetric method. This approach has a linear response to TC between 0.05 µM and 10 µM, with a detection limit of 0.016 µM. In this system, good dispersion provides the substance with a distinct peroxidase activity, which is used to create a colorimetric sensor for detecting TC. Mechanism studies show that the superoxide radical generated by the La-Ce nanomembrane plays a key role in peroxidase catalysis. Finally, the practicality of the method was verified by the determination of TC in food products (milk, pork and honey), which demonstrated that a good recovery rate can be obtained (91.4-102%).

Benchtop NMR Coupled with Chemometrics: A Workflow for Unveiling Hidden Drug Ingredients in Honey-Based Supplements.

Recently, benchtop nuclear magnetic resonance (NMR) spectrometers utilizing permanent magnets have emerged as versatile tools with applications across various fields, including food and pharmaceuticals. Their efficacy is further enhanced when coupled with chemometric methods. This study presents an innovative approach to leveraging a compact benchtop NMR spectrometer coupled with chemometrics for screening honey-based food supplements adulterated with active pharmaceutical ingredients. Initially, fifty samples seized by French customs were analyzed using a 60 MHz benchtop spectrometer. The investigation unveiled the presence of tadalafil in 37 samples, sildenafil in 5 samples, and a combination of flibanserin with tadalafil in 1 sample. After conducting comprehensive qualitative and quantitative characterization of the samples, we propose a chemometric workflow to provide an efficient screening of honey samples using the NMR dataset. This pipeline, utilizing partial least squares discriminant analysis (PLS-DA) models, enables the classification of samples as either adulterated or non-adulterated, as well as the identification of the presence of tadalafil or sildenafil. Additionally, PLS regression models are employed to predict the quantitative content of these adulterants. Through blind analysis, this workflow allows for the detection and quantification of adulterants in these honey supplements.

Antibacterial activity of Hungarian varietal honeys against respiratory pathogens as a function of storage time.

Today, antibiotic therapies that previously worked well against certain bacteria due to their natural sensitivity, are becoming less effective. Honey has been proven to inhibit the biofilm formation of some respiratory bacteria, however few data are available on how the storage time affects the antibacterial effect. The activity of black locust, goldenrod, linden and sunflower honeys from three consecutive years (2020, 2021, 2022) was analyzed in 2022 against Gram-negative (Haemophilus influenzae, H. parainfluenzae, Pseudomonas aeruginosa) and Gram-positive (Streptococcus pneumoniae) bacteria using in vitro microbiological methods. After determining the physicochemical parameters of honey, broth microdilution was applied to determine the minimum inhibitory concentration of each honey type against each bacterium, and crystal violet assay was used to test their antibiofilm effect. The possible mechanism of action was explored with membrane degradation test, while structural changes were illustrated with scanning electron microscopy. Honeys stored for one or two years were darker than fresh honeys, while older honeys had significantly lower antibacterial activity. The most remarkable inhibitory effect was exerted by linden and sunflower honeys, and P. aeruginosa proved to be the most resistant bacterium. Based on our results, honey intended for medicinal purposes should be used as fresh as possible during a treatment.

Effect of various supplements on productive performance of honey bees, in the south Wollo Zone, Ethiopia.

The productivity and well-being of honey bee colonies are greatly influenced by the nutrients present in the hives. A study was conducted to evaluate different supplemental feeds on honey bee productive performance during dearth periods. Thirty colonies were grouped into five (four treatment groups and one control group) and each group contained three sub-groups (2 weak, 2 strong, and 2 very strong). Control groups were not given any supplementation. Treatment diets were T1 (50% sugar syrup + 14% roasted barley powder (beso) + 36% roasted spiced pea powder (Shiro)), T2 (50% powder sugar + 14% white sorghum powder + 36% bakery yeast, T3 (50% powder sugar + 14% white sorghum powder + 36% skimmed milk powder), T4 (50% sugar syrup with infusion of stinging nettle and 1% kerefa + 50% white sorghum powder). Feed was given on the entrance sides. The performance of experimental colonies was measured every 21 days in two phases during the dry season (from 3_2_2021 to 27_4_2021) and the rainy season (from 28-7_2021 to 1_10_2021). Feed intake, space (cm2) of pollen, nectar, and honey in the comb were measured using a frame-sized transparent grid meter. The study revealed significant differences (p<0.0001) in all measured parameters among the various treatments. The diet provided by T4 showed the highest levels of crude protein (18.15%) and carbohydrates (92.15%), whereas the diet presented by T3 had the lowest crude protein content (6.66%) and the diet offered by T1 had the lowest carbohydrate content (61.91%). In general, colonies that received T4 showcased superior performance compared to others. They exhibited a feed intake of 98.3%, a nectar area of 54.3 cm2, a pollen area of 68.7 cm2, a honey area of 311.2 cm2, and a honey yield of 7 kg. Consequently, their net profit amounted to 51.54 USD. On the other hand, the colonies that received T1 had the lowest performance indicators. They demonstrated a feed intake of only 54.7%, a nectar area of 37.6 cm2, a pollen area of 48.8 cm2, a honey area of 254.3 cm2, a honey yield of 2.8 kg, and a net profit of 18.81 USD. The significance of this study was to enable the beekeepers in realizing the effects of feed supplements on the productivity and profitability of honeybee colonies.

Droplet Digital RT-PCR (dd RT-PCR) Detection of SARS-CoV-2 in Honey Bees and Honey Collected in Apiaries across the Campania Region.

Coronaviruses (CoVs), a subfamily of Orthocoronavirinae, are viruses that sometimes present a zoonotic character. Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is responsible for the recent outbreak of COVID-19, which, since its outbreak in 2019, has caused about 774,593,066 confirmed cases and 7,028,881 deaths. Aereosols are the main route of transmission among people; however, viral droplets can contaminate surfaces and fomites as well as particulate matter (PM) in suspensions of natural and human origin. Honey bees are well known bioindicators of the presence of pollutants and PMs in the environment as they can collect a great variety of substances during their foraging activities. The aim of this study was to evaluate the possible role of honey bees as bioindicators of the prevalence SARS-CoV-2. In this regard, 91 samples of honey bees and 6 of honey were collected from different apiaries of Campania region (Southern Italy) in four time periods from September 2020 to June 2022 and were analyzed with Droplet Digital RT-PCR for SARS-CoV-2 target genes Orf1b and N. The screening revealed the presence of SARS-CoV-2 in 12/91 in honey bee samples and in 2/6 honey samples. These results suggest that honey bees could also be used as indicators of outbreaks of airborne pathogens such as SARS-CoV-2.