The Effect of Probiotic Microorganisms on Catalase Activity, Fractional Composition of Soluble Proteins, and Intestinal Microbiota of Honey Bee


  • R.S. Fedoruk Institute of Animal Biology, NAAS of Ukraine, 38 V. Stusa Str., Lviv, 79034, Ukraine
  • I.I. Kovalchuk Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies of Lviv, 50 Pekarska Str., Lviv, 79010, Ukraine
  • A.Z. Pylypets Institute of Animal Biology, NAAS of Ukraine, 38 V. Stusa Str., Lviv, 79034, Ukraine
  • M.M. Tsap Institute of Animal Biology, NAAS of Ukraine, 38 V. Stusa Str., Lviv, 79034, Ukraine
  • Y.V. Lesyk Drohobuch Ivan Franko State Pedagogical University, 24 Ivana Franka Str., Drohobych, 82100, Ukraine
  • R.L. Androshulik Institute of Animal Biology, NAAS of Ukraine, 38 V. Stusa Str., Lviv, 79034, Ukraine
  • O.A. Demchenko Zabolotny Institute of Microbiology and Virology, NAS of Ukraine, 154 Academika Zabolotnoho Str., Kyiv, 03143, Ukraine
  • N.O. Tymoshok Zabolotny Institute of Microbiology and Virology, NAS of Ukraine, 154 Academika Zabolotnoho Str., Kyiv, 03143, Ukraine
  • L.P. Babenko Zabolotny Institute of Microbiology and Virology, NAS of Ukraine, 154 Academika Zabolotnoho Str., Kyiv, 03143, Ukraine



Apis mellifera bees, lactobacilli, catalase, protein fractions, hemolymph, intestinal microbiome spectrum


Recently, there has been a trend toward the use of new effective natural preparations to fight diseases and improve the health of honey bees. It is also known that a well-balanced structure of the intestinal microbiota of honey bees is the basis for their growth, development, strengthening of the immune response, and resistance to infections. It has been established that some strains of lactic acid bacteria that have antibacterial, anti-inflammatory, and immunomodulatory properties, are promising for the development of broad-spectrum probiotic preparations based on them. Therefore, the aim of the work was to determine the effect of probiotic strains Lactobacillus сasei IMV B-7280 and L. plantarum IMV B-7679 on catalase activity, protein content and protein profile of hemolymph, as well as microbiota spectrum of different parts of the intestines of Apis mellifera honey bees. Methods. To conduct the research, a control and two experimental groups of 60-90 bees each were formed. The bees of the control group were fed 60% sugar syrup + 1 mL of distilled H2O for 28 days. The experimental group of bees D1 received 1 mL of 60% sugar syrup + 1 mL of aqueous suspension containing cells of the L. casei IMV B-7280 strain at a concentration of 1 ∙ 106 CFU/mL every day; experimental group of bees D2, in addition to 1 mL of 60% sugar syrup, received 1 mL of aqueous suspension containing cells of L. plantarum IMV B-7976 strain at a concentration of 1 ∙ 104 CFU/mL. Catalase activity of the whole organism tissues was determined using the ability of hydrogen peroxide to form a stable colored complex with molybdenum salts on a spectrophotometer at a wavelength of 410 nm against water. The amount of protein in the whole organism tissues was determined by the Lowry method. The content of total protein in the body of bees was carried out according to the Kjeldahl method. Determination of the content of individual fractions of soluble proteins of the hemolymph was carried out by the method of vertical electrophoresis in a 7.5% polyacrylamide gel. The relative content of protein fractions was determined using the TotalLab TL120 program and expressed as a percentage of the total pool. To determine the qualitative and quantitative spectrum of the gut microbiota of bees, the hindgut and midgut were sampled (separately) from bees of control and experimental groups. The obtained samples were plated on eight selective solid media for cultivation of different groups of microorganisms. Results. A tendency to increase the catalase activity of bee tissues after 28 days of L. casei IMV B-7280 strain use and a consistently higher activity of this enzyme throughout the experimental period under the action of L. plantarum IMV B-7679 strain was established. In the control group of 28th days, the content of bees and catalase activity remained at a constant level. It was shown that on the 14th day and total protein in the body of bees that received L. casei IMV B-7280 strain increased signifi cantly. Water-soluble fractions of hemolymph proteins were found in bees of both groups: γ-globulins, β-globulins, α2-globulins, and α1-globulins. It should be noted that the albumin fraction was not detected. It has been shown that the hindgut contains a much larger number of microorganisms than the midgut. The use of L. casei IMV B-7280 strain led to an increase in the number of lactic acid bacteria and bifidobacteria in both parts of the gut, as well as to a decrease in the number of staphylococci, streptococci, and microscopic fungi. The use of L. plantarum IMV B-7679 strain had a similar effect, but the changes in the composition of gut microbiome were less pronounced. Conclusions. The use of probiotic strains L. casei IMV B-7280 and L. plantarum IMV B-7679 for feeding bees under the conditions of a laboratory thermostat led to quantitative changes in the composition of the intestinal microbiota of bees, namely an increase in the number of lactic acid bacteria and bifidobacteria, as well as a decrease in the number of some other groups of microorganisms in the gut. Probiotic strains stimulated catalase activity of bee’s body tissues, increased the level of total protein, and did not significantly affect the ratio of hemolymph protein fractions.


Download data is not yet available.


Kovalchuk II, Fedoruk RS. [Content of heavy metals in the bees tissues and products depending on agroecological conditions of carpathians region]. The Animal Biology. 2013; 15(4):54-65. Ukrainian.

Neov B, Georgieva A, Shumkova R, Radoslavov G, Hristov P. Biotic and Abiotic Factors Associated with Colonies Mortalities of Managed Honey Bee (Apis mellifera). Diversity. 2019; 11(12):237. DOI:

Almasri H, Tavares DA, Diogon M, et al. Physiological effects of the interaction between Nosema ceranae and sequential and overlapping exposure to glyphosate and difenoconazole in the honey bee Apis mellifera. Ecotoxicol Environ Saf. 2021; 217:112258. DOI:

Balieira KVB, Mazzo M, Bizerra PFV, Guimarães ARJS, Nicodemo D, Mingatto FE. Imidacloprid-induced oxidative stress in honey bees and the antioxidant action of caffeine. Apidologie. 2018; 49:562-572. DOI:

Karavan VV, Kachmaryk DYu, Cherevatov VF, Yazlovytska LS. [Influence of wintering temperature on the state of the antioxidative system in Apis mellifera L.] The Animal Biology. 2021; 23:32-42. Ukrainian. DOI:

Badiou-Bénéteau A, Carvalho SM, Brunet JL, et al. Development of biomarkers of exposure to xenobiotics in the honey bee Apis mellifera: application to the systemic insecticide thiamethoxam. Ecotoxicol Environ Saf. 2012; 82:22-31. DOI:

Jończyk-Matysiak E, Popiela E, Owczarek B, et al. Phages in Therapy and Prophylaxis of American Foulbrood - Recent Implications From Practical Applications. Front Microbiol. 2020; 11:1913. Published 2020 Aug 11. DOI:

Mutinelli F. European legislation governing the authorization of veterinary medicinal products with particular reference to the use of drugs for the control of honey bee diseases. Apiacta. 2003; 38:156-168.

Raymann K, Shaffer Z, Moran NA. Antibiotic exposure perturbs the gut microbiota and elevates mortality in honeybees. PLoS Biol. 2017; 15(3):e2001861. Published 2017 Mar 14. DOI:

Raymann K, Moran NA. The role of the gut microbiome in health and disease of adult honey bee workers. Curr Opin Insect Sci. 2018; 26:97-104. DOI:

Brosi BJ, Delaplane KS, Boots M, de Roode JC. Ecological and evolutionary approaches to managing honeybee disease. Nat Ecol Evol. 2017; 1(9):1250-1262. DOI:

Tauber JP, Collins WR, Schwarz RS, et al. Natural Product Medicines for Honey Bees: Perspective and Protocols. Insects. 2019; 10(10):356. Published 2019 Oct 18. DOI:

Daisley BA, Chmiel JA, Pitek AP, Thompson GJ, Reid G. Missing Microbes in Bees: How Systematic Depletion of Key Symbionts Erodes Immunity. Trends Microbiol. 2020; 28(12):1010-1021. DOI:

Kovalсhuk II, Fedoruk RS, Spivak MYa, Romanovyсh MM, Iskra RYa. Laсtobaсillus сasei IMV B-7280 immunobiotic strain influence on the viability of honey bees and the content of microelements in the organism. Mikrobiol Z. 2021; 83(2):42-50. DOI:

Falalyeyeva TM, Leschenko IV, Beregova TV, Lazarenko LM, Savchuk OM, SichelL M, Tsyryuk OI, Vovk TB, Spivak MYa. Probiotic strains of lactobacilli and bifidobacteria alter pro- and anti-inflammatory cytokines production in rats with monosodium glutamate-induced obesity. Fiziol Zh. 2017; 63(1):17-25. DOI:

Lazarenko LM, Babenko LP, Mokrozub VV, Demchenko OM, Bila VV, Spivak MYa. Effects of oral and vaginal administration of probiotic bacteria on the vaginal microbiota and cytokines production in the case of experimental Staphylococcosis in mice. Mikrobiol Z. 2017; 79(6):105-19. DOI:

Korolyuk MA, Ivanova LI, Mayorova IG, Tokarev BE. [ Method for determining catalase activity]. Laboratory work. 1988; 1:16-18. russian.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with Folin phenol reagent. J Biol Chem. 1951; 193(1):265-275. DOI:

Kjeldahl J. New Method for the Determination of Nitrogen in Organic Substances. Zeitschrift für analytische Chemie. 1883; 22:366-383. DOI:

Laemmli UK. Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature. 1970; 227:680-685. DOI:

[Laboratory animals: breeding, maintenance, use in the experiment]. I.P. Zapadnyuk, editor. K.: Vishcha school, 1983. Russian.

Egorov NS. [Guide to practical exercises in microbiology]. Moscow: Publishing House of Moscow State University. 1999. Russian.

Yazlovitska LS, Kosovan MD, Cherevatov VF, Volkov RA. The catalase activity of Apis mellifera l. upon summer feeding with varying carbohydrate diet. Biological systems. 2016; 8(2):182-188. DOI:

Karavan V, Kachmaryk D, Cherevatov V, Panchuk I, Yazlovytska L. Influence of the summer feeding by carbohydrates on catalase activity in honey bees. Scientific herald of Chernivtsi University. Biology (Biological Systems). 2020; 12(2): 156-165. DOI:

Foelix RF. Biology of Spiders. Frankfurt am Main (Germany): Edition Chimaira; 2015.

Eichelmann MA, Lewbart GA. Hemolymph chemistry reference ranges of the chilean rose tarantula grammostola rosea (walkenaer, 1837) using the vetscan biochemistry analyzer based on IFCC-CLSI C28-A3. J Zoo Wildl Med. 2018; 49(3):528-534. DOI:

Heo J, Kim SJ, Kim JS, Hong SB, Kwon SW. Comparative genomics of Lactobacillus species as bee symbionts and description of Lactobacillus bombintestini sp. nov., isolated from the gut of Bombus ignitus. J Microbiol. 2020; 58(6):445-455. DOI:

Subotic S, Boddicker AM, Nguyen VM, Rivers J, Briles CE, Mosier AC. Honey bee microbiome associated with different hive and sample types over a honey production season. PLoS One. 2019; 14(11):e0223834. Published 2019 Nov 8. DOI:

Corby-Harris V, Maes P, Anderson KE. The bacterial communities associated with honey bee (Apis mellifera) foragers. PLoS One. 2014; 9(4):e95056. Published 2014 Apr 16. DOI:

Alberoni D, Gaggìa F, Baffoni L, Di Gioia D. Beneficial microorganisms for honey bees: problems and progresses. Appl Microbiol Biotechnol. 2016; 100(22):9469-9482. DOI:

Fanciotti MN, Tejerina M, Benítez-Ahrendts MR, Audisio MC. Honey yield of different commercial apiaries treated with Lactobacillus salivarius A3iob, a new bee-probiotic strain. Benef Microbes. 2018; 9(2):291-298. DOI:

Arredondo D, Castelli L, Porrini MP, et al. Lactobacillus kunkeei strains decreased the infection by honey bee pathogens Paenibacillus larvae and Nosema ceranae. Benef Microbes. 2018; 9(2):279-290. DOI:

Truong AT, Kang JE, Yoo MS, et al. Probiotic candidates for controlling Paenibacillus larvae, a causative agent of American foulbrood disease in honey bee. BMC Microbiol. 2023; 23(1):150. Published 2023 May 24. DOI:




How to Cite

Fedoruk, R., Kovalchuk, I., Pylypets, A., Tsap, M., Lesyk, Y., Androshulik, R., Demchenko, O., Tymoshok, N., & Babenko, L. (2023). The Effect of Probiotic Microorganisms on Catalase Activity, Fractional Composition of Soluble Proteins, and Intestinal Microbiota of Honey Bee. Mikrobiolohichnyi Zhurnal, 85(4), 46–57.