Foulbrood Research Articles

Isolation and determination of bacterial microbiota of Varroa destructor and isolation of Lysinibacillus sp. from it

BackgroundThe importance of bees for environmental health is known. Within the scope of this importance, it is of great importance to protect the health of bees and to prevent colony extinction. In this context, it is very important to develop effective methods in combating microorganisms, parasitoids, mites and organisms that cause disease or harm in bees. Both use different methods in terms of bee health.ResultIn this study, the possibility of the bacteria isolated from Varroa destructor mite being bioinsecticide was investigated. Accordingly, six bacteria were isolated from the mite. Isolated bacteria were analyzed according to biochemical tests, molecular analysis, optimum growth pH and phylogenetic tree drawn as Pantoea dispersa (GV1), Lysinibacillus macroides (GV3), Bacillus mycoides (GV4), Lysinibacillus fusiformis (GV5), Pseodomonas lutea (GV5), Lysinibacillus varians (GV7). Lysinibacillus sp. The entomopathogenic feature of Lysinibacillus sp. ranked it as the most important species. When the insecticidal properties of bacteria were examined, they were determined as 53, 90, 62, 95, 74 and 83% for GV1, GV3, GV4, GV5, GV6 and GV7, respectively.ConclusionBased on these results and literature review, Lysinibacillus sp. species had a high potential to be used as bioinsecticide against V. destructor mite.

Effect of population size and Season on the expression of hygienic and aggressive behaviors in lorestan honey bee colonies

Healthy behavior is an appropriate alternative method in combating honey bee pests and diseases to reduce the use of chemical pesticides. In this study, the effect of population size (5, 7 and 10 frames), season (spring and summer) and the interaction between population size and season were investigated on the incidence of health and invasive behaviors of lorestan honey bee colonies. For this purpose, 21 colonies belonging to Lorestan province, Khorramabad city, were used. Liquid nitrogen was used to kill pupae in order to determine the incidence of uncapping and removing traits. In the following, the percentage of uncapping and removing cells was counted 48 hours after pouring liquid nitrogen. The mean of cell removing and uncapping were 94.48%, 92.67%, 91.81% and 88.33% in spring and summer, respectively, but these differences was not significant (p>0.05). The 5 frames populations had the lowest mean of cap uncapping (88.29) and removing (84.43) compared to 7 and 10 frames populations. The incidence of health behavior was affected by the interaction of population size and season so that 7 frame populations had the best health behavior in spring. There was no significant difference between treatments in the occurrence of aggressive behavior. A significant positive correlation was observed between uncapping and removing. In general, the results of this study showed that the lorestan honey bee colonies show good health behavior and they can be used in breeding programs to build resistant populations.

Isolation, characterization, and comparative genomic analysis of vB_PlaP_SV21, new bacteriophage of Paenibacillus larvae

Paenibacillus larvae cause an American foulbrood disease (AFB) that is responsible for the extinction of honeybee colonies and is a honeybee bacterial disease that has to be obligatory notified worldwide. Recently, bacteriophage studies targeting Paenibacillus larvae have emerged as a promising alternative treatment method. The inability of bacteria to create resistance against bacteriophages makes this method advantageous. As a consequence, this study was conducted to describe the genome and biological characteristics of a novel phage capable of lysing Paenibacillus larvae samples isolated from honeybee larva samples in Turkey. The Paenibacillus phage SV21 (vB_PlaP_SV21) was isolated by inducing Paenibacillus larvae strain SV21 with Mitomycin-C. Whole-genome sequencing, comparative genomics, and phylogenetic analysis of vB_PlaP_SV21 were performed. Transmission electron microscopy images showed that vB_PlaP_SV21 phage was a Podovirus morphology. The vB_PlaP_SV21 phage specific for Paenibacillus larvae was determined to belong to the Podoviridae family. Host range and specificity, burst size, lytic activity, and morphological characteristics of the phage were determined. Bioinformatic analysis of the Paenibacillus phage SV21 showed 77 coding sequences in its linear 44,949 bp dsDNA genome with a GC content of 39.33%. In this study, we analysed the genomes of all of the currently sequenced P. larvae phage genomes and classified them into five clusters and a singleton. According to molecular, morphological, and bioinformatics results, ıt was observed that API480 (podovirus), which was reported as a singleton in previous studies and public databases, and Paenibacillus phage SV21 phage could form a new cluster together

Veterinarians and beekeeping: What roles, expectations and future perspectives? - a review paper

Apiculture is an important economy sector, facing a real health crisis. Honeybees, as well as other insect pollinator populations have been in decline over recent decades, and diseases are one of the principal challenges. To face this situation, public and private veterinary services are needed to manage surveillance, control, eradication and prevention measures of honeybee diseases within their territories, in close collaboration with beekeepers and farmers. It is necessary urgently to clarify the current problems and threats to apiculture in the public health sector. Due to possible interactions between bee health and production, the negative effects of environmental xenobiotics and climate change, this field must be considered as an interdisciplinary research issue. Moreover, honeybee veterinary medicine is increasingly engaged in the One Health approach, due to the possibility that the poor health of honeybee colonies affects human life. This review identified the key veterinarian roles, gaps in their higher education curriculum, as well as the necessity for research linkage between expectations and professional perspectives.

Upconversion-Linked Immunoassay for the Diagnosis of Honeybee Disease American Foulbrood

American foulbrood (AFB) caused by the bacterium Paenibacillus larvae is the most destructive disease of the honeybee brood. Therefore, rapid and sensitive detection methods are required to limit spreading of this pathogen, which has a major impact on agriculture and biodiversity. While P. larvae is typically detected by microbial cultivation or polymerase chain reaction, antibody-based detection represents a viable alternative. Here, we prepared an antibody specific for P. larvae and used it for the development of an upconversion-linked immunosorbent assay (ULISA). Photon-upconversion nanoparticles (UCNP) were conjugated to streptavidin via a PEG-linker using copper-catalyzed click chemistry to replace the enzyme label in conventional enzyme-linked immunosorbent assay (ELISA). The ULISA showed low cross-reactivity and provided a limit of detection of 2.9 × 10 <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">3</sup> CFU/mL, representing a 22-fold improvement compared to the ELISA. This level is within the bacterial loads present in honeybee larvae during an AFB infection. The assay was successfully applied to the analysis of spiked samples of bees, larvae, and hive debris.

Nosema Disease of European Honey Bees

Nosematosis is currently a frequently discussed honey bee disease caused by two types of Microsporidia: Nosema apis and Nosema ceranae. Nosematosis as an intestinal disease caused by these species is one of the main factors associated with the weakening and loss of hives, with none of the stressors acting in isolation and all having an important synergistic or additive effect on the occurrence of parasitic infection. The most important factors are exposure to pesticides and nutritional stress, both worsening the immune response. Honey bees Apis mellifera become more susceptible to parasites and subsequently the disease manifests itself. Choosing the right laboratory diagnostics is important to determine the prevalence of both species. Our review summarizes the most commonly used methodologies, especially polymerase chain reaction (PCR), which is a reliable method for detecting nosematosis, as well as for distinguishing between the two species causing the disease.

Effects of Insecticides and Microbiological Contaminants on Apis mellifera Health.

Over the past two decades, there has been an alarming decline in the number of honey bee colonies. This phenomenon is called Colony Collapse Disorder (CCD). Bee products play a significant role in human life and have a huge impact on agriculture, therefore bees are an economically important species. Honey has found its healing application in various sectors of human life, as well as other bee products such as royal jelly, propolis, and bee pollen. There are many putative factors of CCD, such as air pollution, GMO, viruses, or predators (such as wasps and hornets). It is, however, believed that pesticides and microorganisms play a huge role in the mass extinction of bee colonies. Insecticides are chemicals that are dangerous to both humans and the environment. They can cause enormous damage to bees’ nervous system and permanently weaken their immune system, making them vulnerable to other factors. Some of the insecticides that negatively affect bees are, for example, neonicotinoids, coumaphos, and chlorpyrifos. Microorganisms can cause various diseases in bees, weakening the health of the colony and often resulting in its extinction. Infection with microorganisms may result in the need to dispose of the entire hive to prevent the spread of pathogens to other hives. Many aspects of the impact of pesticides and microorganisms on bees are still unclear. The need to deepen knowledge in this matter is crucial, bearing in mind how important these animals are for human life.

Environmental exposures associated with honey bee health

Bee health is declining on a global scale, yet the exact causes and their interactions responsible for the decline remain unknown. To more objectively study bee health, recently biomarkers have been proposed as an essential tool, because they can be rapidly quantified and standardized, serving as a comparable measure across bee species and varying environments. Here, we used a systems biology approach to draw associations between endogenous and exogenous chemical profiles, with pesticide exposure, or the abundance of the 21 most common honey bee diseases. From the analysis we identified chemical biomarkers for both pesticide exposure and bee diseases along with the mechanistic biological pathways that may influence disease onset and progression. We found a total of 2352 chemical features, from 30 different hives, sampled from seven different locations. Of these, a total of 1088 significant associations were found that could serve as chemical biomarker profiles for predicting both pesticide exposure and the presence of diseases in a bee colony. In almost all cases we found novel external environmental exposures within the top seven associations with bee diseases and pesticide exposures, with the majority having previously unknown connections to bee health. We highlight the exposure-outcome paradigm and its ability to identify previously uncategorized interactions from different environmental exposures associated with bee diseases, pesticides, mechanisms, and potential synergistic interactions of these that are responsible for honey bee health decline.

American Foulbrood in the Czech Republic: ERIC II Genotype of Paenibacillus Larvae Is Prevalent.

American foulbrood (AFB) is a dangerous disease of honeybees (Apis mellifera) caused by the spore-forming bacterium Paenibacillus larvae. According to the ERIC (enterobacterial repetitive intergenic consensus) classification, five genotypes are distinguished, i.e., I, II, III, IV, and V, which differ in their virulence and prevalence in colonies. In the Czech Republic, AFB prevalence is monitored by the State Veterinary Administration; however, the occurrence of specific P. larvae genotypes within the country remains unknown. In this study, our aim was to genotype field P. larvae strains collected in the Czech Republic according to the ERIC classification. In total, 102 field isolates from colonies with AFB clinical symptoms were collected from various locations in the Czech Republic, and the PCR genotypization was performed using ERIC primers. We confirmed the presence of both ERIC I and II genotypes, while ERIC III, IV, and V were not detected. The majority of samples (n = 82, 80.4%) were identified as ERIC II, while the ERIC I genotype was confirmed only in 20 samples (19.6%). In contrast to other European countries, the ERIC II genotype is predominant in Czech honeybee colonies. The ERIC I genotype was mostly detected in border regions close to Poland, Slovakia, and Austria.

Beekeeping practices and challenges at Lemo District, Hadiya Zone, Southern Ethiopia

Beekeeping has been an integral part of the Ethiopian agriculture and yielding 1.3% of agricultural GDP. Hence, the study was aimed to evaluate the purpose of beekeeping and major constraints impeding beekeeping practices at Lemo District, Hadiya Zone, Southern Ethiopia. Questionnaires were organized to collect survey data. A multi-stage purposive sampling technique was used and one hundred forty respondents (140 beekeepers) were interviewed using a semi-structured questionnaire. Key informant interviews and Focus group discussion to support the interpretation of the interview data. The data were organized and analyzed using one-way analysis of variance via Statistical Package for Social Sciences. Accordingly, beekeeping was mostly carried out using traditional hives. From beekeeper’s respondents 80%, are involved in beekeeping mainly for both income and household consumption. Beekeepers use income obtained from beekeeping for buying crop inputs, payment for annual land taxation, and purchase food grain. The mean honey yield of traditional log hives, transitional top-bar hives, and modern hives with frames were 5.25 ± 0.38, 7.52 ± 0.54, and 12.78 ± 0.56 kg per hive per year, respectively. The most hampering factors for honey bee production were honey bee diseases, improper pesticide application, and shortage of bee forage. The finding of this study concluded that beekeeping was mostly carried out using traditional hives. Hence, well-organized training should be given for the beekeepers to practice internal colony inspection and improved beekeeping technologies via a full package program. Efforts should be geared to alleviate constraints like honey bee diseases, improper application of pesticides, and bee forage shortage hampering beekeeping in the district.

Study on Major Parasitic Diseases of Adult Honeybees in Three Districts of Kaffa Zone, Southern Ethiopia.

Results Out of 384 samples, the overall prevalence of small hive beetle, Nosema apis, and amoeba (Malpighamoeba mellificae) diseases was 39%, 45.3%, and 40.3%, respectively. The overall prevalence of these three diseases was ascertained, with a statistically significant (p < 0.05) variation in the overall prevalence of N. apis disease and amoeba (M. mellificae) disease between hive type and agroecologies. Conclusion The research indicates that N. apis, M. mellificae, and small hive beetle are the major parasites that affect honeybees in the study area. In order to classify and describe honeybee diseases and pests associated with Ethiopia's local honeybees, several field diagnostic surveys and laboratory research works need to be performed.

Biologically Active Extracts from Different Medicinal Plants Tested as Potential Additives against Bee Pathogens.

Honey bees (Apis mellifera) perform pollination service for many agricultural crops and contribute to the global economy in agriculture and bee products. However, honey bee health is an ongoing concern, as illustrated by persistent local population decline, caused by some severe bee diseases (e.g., nosemosis, AFB, EFB, chalkbrood). Three natural recipes are in development based on the bioactive compounds of different plants extract (Agastache foeniculum, Artemisia absinthium, Evernia prunastri, Humulus lupulus, Laurus nobilis, Origanum vulgare and Vaccinium myrtillus), characterised by HPLC-PDA. The antimicrobial activity of these recipes was tested in vitro against Paenibacillus larvae, Paenibacillus alvei, Brevibacillus laterosporus, Enterococcus faecalis, Ascosphaera apis and in vivo against Nosema ceranae. A mix of 20% blueberry, 40% absinthium, 10% oakmoss, 10% oregano, 10% Brewers Gold hops, 5% bay laurel and 5% anise hyssop extract showed the strongest antibacterial and antifungal activity. Combing several highly active plant extracts might be an alternative treatment against bee-disease-associated parasites and pathogens, in particular to replace synthetic antibiotics.

Conducting an International, Exploratory Survey to Collect Data on Honey Bee Disease Management and Control

The Istituto Zooprofilattico Sperimentale del Lazio e della Toscana (IZSLT) and the International Federation of Beekeepers’ Associations (Apimondia) set up a worldwide survey in 2015–2017 to gather information on beekeepers’ perceptions concerning good beekeeping practices, the main honey bee diseases, and the technical assistance they receive. The on-line dissemination of the survey was facilitated by the “Technologies and Practices for Small Agricultural Producers” platform (TECA) of the Food and Agriculture Organization of the United Nations (FAO). In total, 248 questionnaires were received from the European region and the results are reported here. Varroosis remains the biggest concern and is the most important reason for beekeepers’ use of veterinary medicines. However, a sustainable approach to managing Varroa was detected. American foulbrood (AFB) and European foulbrood (EFB) infections are mainly managed through the shook swarm technique or burning the hives. Concerning technical assistance for disease management, beekeepers were mainly supported by their associations or expert beekeepers. Relevant data were collected and analyzed but information from many low-income countries in Africa or Asia is still missing, and more efforts are needed to fill the knowledge gaps.

THE RESULTS OF THE USE OF BIOTECHNOLOGICAL DRUGS IN VETERINARY MEDICINE, PROMISING FOR MEDICINE (PART 1)

Cell and serum nucleases are considered as a natural biological barrier, one of the first links in the body’s antiviral defense: nucleases act on viruses that the usual immunological barrier cannot resist. Inactivation of the enzymatic properties of nucleases also leads to the loss of their antiviral activity. It was found that the nucleases do not inactivate the native virus outside the cell and act mainly on the virus that multiplies in the cells. There is a direct dependence of the antiviral activity of endonuclease on the concentration of the enzyme in the medium. The first antiviral drug created on the basis of the Serratia marcescens endonuclease was called bacterial endonuclease. Its technology was developed by joint research of NIKTI BAS of the Ministry of Medical Industry (Berdsk, Novosibirsk region) and ICIG SB of the USSR Academy of Sciences in 1973–1984. Serratia marcescens endonuclease is capable of cleaving the nucleic acids of both RNA and DNA-containing viruses. The enzyme inhibits the reproduction of vesicular stomatitis and smallpox vaccine viruses in chicken fibroblast cell culture, respectively. It was the first antiviral drug in the history of beekeeping, designed to prevent acute and chronic paralysis and other viral diseases of bees. The drug was also intended as an antiviral agent for a wide range of diseases in various organisms. The effectiveness of bacterial endonuclease as a means of prevention and treatment of respiratory viral diseases in calves was studied. The research was carried out at the industrial complex for growing heifers of the Gosnensky state farm in the Leningrad region on calves of a black-and-white breed of 20–60 days old. For the experiments, we used bacterial endonuclease produced by the Vyshnevolotsky Plant of Enzyme Preparations or NICTI BAS of the Ministry of Medical Industry. In the future, NIKTI BAV together with Diafarm LLC developed a second – generation antiviral drug, endoglucin, based on bacterial endonucleosis. Production experiments to study its effect as a therapeutic and prophylactic agent for respiratory diseases of calves were conducted during 2007– 2011. in the agricultural complex «Prichulymsky» of the Achinsky district of the Krasnoyarsk Territory with the participation of the Department of Epizootology and Parasitology of the Faculty of Veterinary Medicine of KrasGAU and in CJSC «Suzdalskoye» of the Dovolensky district of the Novosibirsk region. Prophylactic use of endoglucin in calves with bronchopneumonia can reduce the incidence from 2.2 to 3.1 times. The drug endoglucin in combination with drugs indicated for the treatment of pulmonary pathology, has therapeutic effectiveness in bronchopneumonia of calves.

Diversity of aerobic spore-forming bacteria isolated from fresh bee pollen intended for human consumption in Argentina

Bee pollen is the result of the agglutination of pollen grains collected from flowers and mixed with nectar and salivary secretions by honey bees. Bee pollen is a natural product exposed to environmental conditions and also provides a unique microhabitat for yeasts and bacterial communities. We analyzed 30 fresh bee pollen samples obtained from the main producing areas of Argentina to identify aerobic-spore-forming bacteria. We obtained 73 isolates belonging to 16 different species through isolation on selective and differential media, morphological and biochemical tests, and PCR and RFLP analysis of genes encoding 16S rRNA. Our data revealed that Bacillus cereus sensu stricto was the most predominant species (50%), followed by Bacillus megaterium (40%) and Bacillus subtilis (40%), respectively. In a minor proportion, Paenibacillus polymyxa (20%), Paenibacillus larvae (17%), Bacillus pumilus (13%), Bacillus licheniformis (13%), Bacillus amyloliquefaciens (10%), Lysinibacillus sphaericus (7%), Bacillus coagulans (7%), Rummelliibacillus stabekisii (7%), Bacillus thuringiensis (7%), Bacillus clausii (3%), Paenibacillus alvei (3%), Bacillus simplex (3%), and Paenibacillus amylolyticus (3%) were also found. Our results showed that Argentinean bee pollen could transmit honey bee diseases due to the presence of viable spores of P. larvae and also spores of toxicogenic B. cereus s.s. and B. megaterium strains.

When a Tritrophic Interaction Goes Wrong to the Third Level: Xanthoxylin From Trees Causes the Honeybee Larval Mortality in Colonies Affected by the River Disease.

The "River Disease" (RD), a disorder impacting honeybee colonies located close to waterways with abundant riparian vegetation (including Sebastiania schottiana, Euphorbiaceae), kills newly hatched larvae. Forager bees from RD-affected colonies collect honeydew excretions from Epormenis cestri (Hemiptera: Flatidae), a planthopper feeding on trees of S. schottiana. First-instar honeybee larvae fed with this honeydew died. Thus, we postulated that the nectars of RD-affected colonies had a natural toxin coming from either E. cestri or S. schottiana. An untargeted metabolomics characterization of fresh nectars extracts from colonies with and without RD allowed to pinpoint xanthoxylin as one of the chemicals present in higher amounts in nectar from RD-affected colonies than in nectars from healthy colonies. Besides, xanthoxylin was also found in the aerial parts of S. schottiana and the honeydew excreted by E. cestri feeding on this tree. A larva feeding assay where xanthoxylin-enriched diets were offered to 1st instar larvae showed that larvae died in the same proportion as larvae did when offered enriched diets with nectars from RD-colonies. These findings demonstrate that a xenobiotic can mimic the RD syndrome in honeybee larvae and provide evidence of an interspecific flow of xanthoxylin among three trophic levels. Further, our results give information that can be considered when implementing measures to control this honeybee disease.

Treatment of bee families without drugs, or a zootechnical method of combating diseases of bees

Todayall the monographs, recommendations, instructions, and technologies after the successful experiments on the treatment of bees against European spot disease with penicillin, there are instructions from the NIP (Research Institute of Prevention) to use antibiotics to the treatment of all bee diseases, but especially in the form of prophylactic supplementation. Many antibiotics (chloramphenicol) and nitrofuran-based medicines are now being produced. In addition, all new medications are intensively recommended in all issues of Beekeeping magazine. These drugs are often newly invented, untested remedies that are advertised as a panacea for all diseases. Antibiotics are introduced into honey and bee-bees in the treatment and prevention of bee diseases. When such products are consumed, people develop various diseases with severe and even incurable consequences. With this in mind, the European Union and the United States do not buy and even strictly prohibit honey from China, Brazil, Argentina and Turkey across borders. The Department of Biology, Bioresources and Aquaculture at Novosibirsk State Agrarian University has for the first time developed a technology for keeping bees without the use of drugs. The long-term use of this technology in the apiaries of Western Siberia has made it possible to achieve good preservation of bee families and high honey yields. In addition, the cost of beekeeping products has fallen dramatically, as farms have stopped spending huge sums on purchasing medicines. Scientists have improved and successfully applied heat treatment of bee colonies to combat varroaatosis, nosematosis and viral diseases.

Apicultural practice and disease prevalence in Apis mellifera, New Zealand: a longitudinal study

In New Zealand, the introduced honey bee (Apis mellifera) is a valuable production animal, providing pollination services for horticultural crops and significant export volumes of honey, especially mānuka honey. Honey bees in New Zealand are free from a number of significant diseases and pests such as European foulbrood, acarine disease, small hive beetle, Israeli acute paralysis virus and tropilaelaps mites. We sought to determine the health status of honey bees in New Zealand using a longitudinal study that followed 60 beekeepers over 2.5 years, ascertaining disease and pest status in their elected study apiary and interviewing them every spring and autumn. Participant beekeepers accounted for the management of approximately 12% of the beehives registered in New Zealand. Differences in beekeeping practices were observed between the North Island and the South Island. Nosema ceranae was found almost exclusively on the North Island and did not displace Nosema apis over the course of the study. Lotmaria passim showed a reverse-phase seasonality to nosema, peaking in autumn at near 100% prevalence. The prevalence of Varroa in apiaries varied seasonally between 45.0% and 46.7% in spring and between 65.0% and 69.5% in autumn, with most infestation rates below 3 mites per 100 bees. The detection rate of symptomatic American foulbrood disease during our hive inspections was very low, between 0.00% and 0.85% hive-level prevalence dependent on the season. This study sets a foundation for understanding honey bee health in New Zealand.

Detection Bioactive Metabolites of Fructobacillus fructosus Strain HI-1 Isolated from Honey Bee's Digestive Tract Against Paenibacillus larvae.

American foulbrood is a devastating disease of honey bee, causing economic loss in the beekeeping industry. The disease mainly causes reduction in honeybee populations which negatively affect the honey bee's major role as natural pollinators of significant crops and wildflowers. Thus, it is crucial to develop safe efficient strategies to control the disease and to improve bee colony health. Using lactic acid bacteria (LAB) as an alternative to chemical treatments is a promising novel technique for tackling honeybee diseases and improving their immunity. The endogenous LAB isolates were recovered from honeybee gut samples collected from different apiaries in two Egyptian governorates and screened for antagonistic activities against Paenibacillus larvae (pathogen of AFB disease). The results showed that 53.3% of tested LAB isolates (n = 120) exhibited antagonistic activities against P. larvae. The minimum inhibitory concentration and minimum bactericidal concentration of the most potent LAB isolate (with an inhibition zone of 44 mm) were 100 and 125 µL/mL, respectively. 16S rRNA sequencing identified the most potent isolate as Fructobacillus fructosus HI-1. The bioactive metabolites of F. fructosus were extracted with ethyl acetate and fractionated on thin-layer chromatography (TLC); also, bioactive fractions were detected. Heptyl 2-methylbutyrate, di-isobutyl phthalate, D-turanose, heptakis (trimethylsilyl), di-isooctyl phthalate, and hyodeoxycholic acid compounds were identified in the bioactive fractions. The result explores the promising administration of probiotic metabolites to control honeybee AFB disease, as a natural tool to substitute antibiotics and chemicals in disease-controlling strategies.

Scientific justification of the methods on disinfection and sanitation in beekeeping

There are no specific protection means (vaccines) in beekeeping. To control infectious diseases of bees, various medicinal preparations are used: antibiotics, sulfonamides, bacteriophages, but they do not lead to the improvement of apiaries without disinfection. Disinfection preparations for the disinfection of beekeeping facilities according to the current situation are approved for use by developers and manufacturers themselves. Often, the proposed preparations for disinfection of beekeeping facilities are studied with large methodological errors, which leads to negative consequences for effectiveness of health measures. In order to eliminate methodological errors, methods have been developed for determining the nature and amount of microbial protection on test objects, the procedure for applying the disinfectant solution and its consumption per 1 m2 of surface, the method and procedure for taking a sample from the disinfected surface, neutralizing the disinfectant, determining the concentration of non-disinfected microorganisms. The use of developed methods in testing of new disinfectants allows to obtain reliable results of disinfection of various objects of beekeeping.