Skip to main content Accessibility help

Piperine as a new natural supplement with beneficial effects on the life-span and defence system of honeybees

  • M. Schulz (a1), A. Łoś (a1) (a2), M. Grzybek (a3) (a4), R. Ścibior (a3) and A. Strachecka (a1)...


Many factors, including pathogens, environmental change and breeding techniques, affect honeybee immunity/resistance, so substances and natural supplements that enhance it are desired. To eliminate the impact of unknown external factors, in 2016 a cage experiment was conducted under constant laboratory conditions (35 °C, 65% relative humidity). Bees in the control group were fed with sugar dissolved in water at ratio 1:1 ad libitum with no additives, while the other group was fed with sugar syrup (1:1) supplemented with piperine (3 µg/ml) ad libitum. The piperine-treated workers lived 9 days longer compared to the control group. In the piperine-consuming group, protein concentration and the activities of antioxidative enzymes, such as superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) and glutathione S-transferase (GST), were higher than in the control group. The activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) were also higher in the piperine-treated group. Neutral and acidic proteases inhibitors, as well as neutral protease activities, were higher in the haemolymph of the piperine-treated workers than in untreated bees. Acidic protease activities in the haemolymph were higher in untreated workers only on days 18 and 32. Alkaline protease activities in the control bees were higher from day 10. From 10 days old, the total antioxidant capacity level was significantly higher in the haemolymph of piperine-treated workers. Piperine decreased DNA methylation levels significantly in the older bees. The compound could have the potential to be a natural diet supplement increasing apian resistance to stress factors.


Corresponding author

Author for correspondence: A. Strachecka, E-mail:


Hide All
Aebi, H (1984) Catalase in vitro. Methods in Enzymology 105, 121126.
Alaux, C, Dantec, C, Parrinello, H and Le Conte, Y (2011) Nutrigenomics in honey bees: digital gene expression analysis of pollen's nutritive effects on healthy and varroa-parasitized bees. BMC Genomics 12, article no. 496. doi: 10.1186/1471-2164-12-496.
Al-Lawati, HT, Al-Ajmi, HIS and Waly, MI (2018) Antioxidant and health properties of beehive products against oxidative stress-mediated carcinogenesis. In Waly, MI and Rahman, MS (eds), Bioactive Components, Diet and Medical Treatment in Cancer Prevention. Cham, Switzerland: Springer, pp. 97103.
Anson, ML (1938) The estimation of pepsin, trypsin, papain and cathepsin with hemoglobin. Journal of General Physiology 22, 7984.
Aurori, CM, Buttstedt, A, Dezmirean, DS, Mărghitaş, LA, Moritz, RF and Erler, S (2014) What is the main driver of ageing in long-lived winter honeybees: antioxidant enzymes, innate immunity, or vitellogenin? Journal of Gerontology. Series A, Biological Sciences And Medical Sciences 69, 633639.
Badmaev, V, Majeed, M and Prakash, L (2000) Piperine derived from black pepper increases the plasma levels of coenzyme Q10 following oral supplementation. Journal of Nutritional Biochemistry 11, 109113.
Bajda, M, Łoś, A and Merska, M (2014) Effect of amphotericin B on the biochemical markers in the haemolymph of honey bees. Medycyna Weterynaryjna (Veterinary Medicine) 70, 766769.
Benzie, IF and Strain, JJ (1996) The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Analytical Biochemistry 239, 7076.
Bhardwaj, RK, Glaeser, H, Becquemont, L, Klotz, U, Gupta, SK and Fromm, MF (2002) Piperine, a major constituent of black pepper, inhibits human P-glycoprotein and CYP3A4. Journal of Pharmacology and Experimental Therapeutics 302, 645650.
Bukhari, IA, Pivac, N, Alhumayyd, MS, Mahesar, AL and Gilani, AH (2013) The analgesic and anticonvulsant effects of piperine in mice. Journal of Physiology and Pharmacology 64, 789794.
Bull, JC, Ryabov, EV, Prince, G, Mead, A., Zhang, C, Baxter, LA, Pell, JK, Osborne, JL and Chandler, D (2012) A strong immune response in young adult honeybees masks their increased susceptibility to infection compared to older bees. PLoS Pathogens 8, e1003083.
Cabiscol, E, Tamarit, J and Ros, J (2000) Oxidative stress in bacteria and protein damage by reactive oxygen species. International Microbiology 3, 38.
Chance, B and Maehly, AC (1955) Assay of catalases and peroxidases. Methods in Enzymology 2, 764775.
Chopra, B, Dhingra, AK, Kapoor, RP and Prasad, DN (2016) Piperine and its various physicochemical and biological aspects: a review. Open Chemistry Journal 3, 7596.
Deng, Y, Sriwiriyajan, S, Tedasen, A, Hiransai, P and Graidist, P (2016) Anti-cancer effects of Piper nigrum via inducing multiple molecular signaling in vivo and in vitro. Journal of Ethnopharmacology 188, 8795.
Dhule, SS, Penfornis, P, Frazier, T, Walker, R, Feldman, J, Tan, G, He, J, Alb, A, John, V and Pochampally, R (2012) Curcumin-loaded γ-cyclodextrin liposomal nanoparticles as delivery vehicles for osteosarcoma. Nanomedicine: Nanotechnology, Biology, and Medicine 8, 440451.
Do, MT, Kim, HG, Choi, JH, Khanal, T, Park, BH, Tran, TP, Jeong, TC and Jeong, HG (2013) Antitumor efficacy of piperine in the treatment of human HER2-overexpressing breast cancer cells. Food Chemistry 141, 25912599.
Donkersley, P, Rhodes, G, Pickup, RW, Jones, KC, Power, EF, Wright, GA and Wilson, K (2017) Nutritional composition of honey bee food stores vary with floral composition. Oecologia 185, 749761.
Farjan, M, Dmitryjuk, M, Lipiński, Z, Łopieńska-Biernat, E and Żółtowska, K (2012) Supplementation of the honey bee diet with vitamin C: the effect on the antioxidative system of Apis mellifera carnica brood at different stages. Journal of Apicultural Research 51, 263270.
Filipiak, M, Kuszewska, K, Asselman, M, Denisow, B, Stawiarz, E, Woyciechowski, M and Weiner, J (2017) Ecological stoichiometry of the honeybee: Pollen diversity and adequate species composition are needed to mitigate limitations imposed on the growth and development of bees by pollen quality. PLoS ONE 12, article no. e0183236.
Foret, S, Kucharski, R, Pittelkow, Y, Lockett, G and Maleszka, R (2009) Epigenetic regulation of the honey bee transcriptome: unravelling the nature of methylated genes. BMC Genomics 10, article no. 472.
Fries, I, Chauzat, MP, Chen, YP, Doublet, V, Genersch, E, Gisder, S, Higes, M, McMahon, DP, Martín-Hernández, R, Natsopoulou, M, Paxton, RJ, Tanner, G, Webster, TC and Williams, GR (2013) Standard methods for Nosema research. Journal of Apicultural Research 52, 128.
Fünfhaus, A, Göbel, J, Ebeling, J, Knispel, H, Garcia-Gonzalez, E and Genersch, E (2018) Swarming motility and biofilm formation of Paenibacillus larvae, the etiological agent of American Foulbrood of honey bees (Apis mellifera). Scientific Reports 8, article no. 8840.
Glavinic, U, Stankovic, B, Draskovic, V, Stevanovic, J, Petrovic, T, Lakic, N and Stanimirovic, Z (2017) Dietary amino acid and vitamin complex protects honey bee from immunosuppression caused by Nosema ceranae. PLoS ONE 12, article no. e0187726.
Goulson, D, Nicholls, E, Botías, C and Rotheray, EL (2015) Bee declines driven by combined stress from parasites, pesticides, and lack of flowers. Science 347, article no. 1255957. DOI: 10.1126/science.1255957.
Griesch, J and Vilcinskas, A (1998) Proteases released by entomopathogenic fungi impair phagocytic activity, attachment and spreading of plasmatocytes isolated from haemolymph of the greater wax moth Galleria mellonella. Biocontrol Science and Technology 8, 517531.
Grzywnowicz, K, Ciołek, A, Tabor, A and Jaszek, M (2009) Profiles of the body-surface proteolytic system of honey bee queens, workers and drones: Ontogenetic and seasonal changes in proteases and their natural inhibitors. Apidologie 40, 419.
Johnson, RM, Ellis, MD, Mullin, CA and Frazier, M (2010) Pesticides and honey bee toxicity – USA. Apidologie 41, 312331.
Kapoor, IPS, Singh, B, Singh, G, De Heluani CS, D, Lampasona, MP and Catalan, CAN (2009) Chemistry and in vitro antioxidant activity of volatile oil and oleoresins of black pepper (Piper nigrum). Journal of Agricultural and Food Chemistry 57, 53585364.
Kevan, PG and Viana, BF (2003) The global decline of pollination services. Biodiversity 4, 38.
Kumar, A, Khan, IA, Koul, S, Koul, JL, Taneja, SC, Ali, I, Ali, F, Sharma, S, Mirza, ZM, Kumar, M, Sangwan, PL, Gupta, P, Thota, N and Qazi, GN (2008) Novel structural analogues of piperine as inhibitors of the NorA efflux pump of Staphylococcus aureus. Journal of Antimicrobial Chemotherapy 61, 12701276.
Lee, TM and Lin, YH (1995) Trypsin inhibitor and trypsin-like protease activity in air- or submergence-grown rice (Oryza sativa L.) coleoptiles. Plant Science 106, 4354.
Lee, SH, Kim, HY, Back, SY and Han, HK (2018) Piperine-mediated drug interactions and formulation strategy for piperine: Recent advances and future perspectives. Expert Opinion on Drug Metabolism & Toxicology 14, 4357.
Li, C, Xu, B, Wang, Y, Yang, Z and Yang, W (2014) Protein content in larval diet affects adult longevity and antioxidant gene expression in honey bee workers. Entomologia Experimentalis et Applicata 151, 1926.
Li, Q, Zhai, W, Jiang, Q, Huang, R, Liu, L, Dai, J, Gong, W, Du, S and Wu, Q (2015) Curcumin-piperine mixtures in self-microemulsifying drug delivery system for ulcerative colitis therapy. International Journal of Pharmaceutics 490, 2231.
Łoś, A and Strachecka, A (2018) Fast and cost-effective biochemical spectrophotometric analysis of solution of insect “blood” and body surface elution. Sensors 18, article no. 1494. doi: 10.3390/s18051494.
Lyko, F and Maleszka, R (2011) Insect as innovative models for functional studies of DNA methylation. Trends in Genetics 27, 127164.
Matysiak, J, Dereziński, P, Klupczyńska, A, Matysiak, J, Kaczmarek, E and Kokot, ZJ (2014) Effects of a honeybee sting on the serum free amino acid profile in humans. PLoS One 9, article no. e103533. doi:10.1371/journal.pone.0103533.
McNamara, FN, Randall, A and Gunthorpe, MJ (2005) Effects of piperine, the pungent component of black pepper, at the human vanilloid receptor (TRPV1). British Journal of Pharmacology 144, 781790.
Mehta, A, Kaur, G and Chintamaneni, M (2012) Piperine and quercetin enhances antioxidant and hepatoprotective effect of curcumin in paracetamol induced oxidative stress. International Journal of Pharmacology 8, 101107.
Mittal, R and Gupta, RL (2000) In vitro antioxidant activity of piperine. Methods and Findings in Experimental and Clinical Pharmacology 22, 271274.
Münch, D, Amdam, GV and Wolschin, F (2008) Ageing in a eusocial insect: molecular and physiological characteristics of life span plasticity in the honey bee. Functional Ecology 22, 407421.
Nahak, G and Sahu, RK (2011) Phytochemical evaluation and antioxidant activity of Piper cubeba and Piper nigrum. Journal of Applied Pharmaceutical Science 1, 153157.
O'Neal, ST, Anderson, TD and Wu-Smart, JY (2018) Interactions between pesticides and pathogen susceptibility in honey bees. Current Opinion in Insect Science 26, 5762.
Ouyang, DY, Zeng, LH, Pan, H, Xu, LH, Wang, Y, Liu, KP and He, XH (2013) Piperine inhibits the proliferation of human prostate cancer cells via induction of cell cycle arrest and autophagy. Food and Chemical Toxicology 60, 424430.
Paarakh, PM, Sreeram, DC, Shruthi, SD and Ganapathy, SP (2015) In vitro cytotoxic and in silico activity of piperine isolated from Piper nigrum fruits Linn. In Silico Pharmacology 3, article no. 9. doi:10.1186/s40203-015-0013-2.
Parker, R, Guarna, MM, Melathopoulos, AP, Moon, KM, White, R, Huxter, E, Pernal, SF and Foster, LJ (2012) Correlation of proteome-wide changes with social immunity behaviors provides insight into resistance to the parasitic mite, Varroa destructor, in the honey bee (Apis mellifera). Genome Biology 13, article no. R81. doi: 10.1186/gb-2012-13-9-r81.
Patial, V, Sukapaka, M, Sharma, S, Pratap, K, Singh, D and Padwad, YS (2015) Synergistic effect of curcumin and piperine in suppression of DENA-induced hepatocellular carcinoma in rats. Environmental Toxicology and Pharmacology 40, 445452.
Pawlowski, AC, Westman, EL, Koteva, K, Waglechner, N and Wright, GD (2018) The complex resistomes of Paenibacillaceae reflect diverse antibiotic chemical ecologies. The ISME Journal 12, 885897.
Podczasy, JJ and Wei, R (1988) Reduction of iodonitrotetrazolium violet by superoxide radicals. Biochemical and Biophysical Research Communications 150, 12941301.
Ptaszyńska, AA, Gancarz, M, Hurd, PJ, Borsuk, G, Wiącek, D, Nawrocka, A, Strachecka, A, Załuski, D and Paleolog, J (2018 a) Changes in the bioelement content of summer and winter western honeybees (Apis mellifera) induced by Nosema ceranae infection. PLoS ONE 13, article no. e0200410.
Ptaszyńska, AA, Trytek, M, Borsuk, G, Buczek, K, Rybicka-Jasińska, K and Gryko, D (2018 b) Porphyrins inactivate Nosema spp. microsporidia. Scientific Reports 8, article no. 5523.
Rada, V, Machova, M, Huk, J, Marounek, M and Duskova, D (1997) Microflora in the honeybee digestive tract: counts, characteristics and sensitivity to veterinary drugs. Apidologie 28, 357365.
Raymann, K, Motta, EVS, Girard, C, Riddington, IM, Dinser, JA and Moran, NA (2018) Imidacloprid decreases honey bee survival rates but does not affect the gut microbiome. Applied and Environmental Microbiology 84, article no. e00545-18. doi: 10.1128/AEM.00545-18.
Reddy, CA, Somepalli, V, Golakoti, T, Kanugula, AK, Karnewar, S, Rajendiran, K, Vasagiri, N, Prabhakar, S, Kuppusamy, P, Kotamraju, S and Kutala, VK (2014) Mitochondrial-targeted curcuminoids: a strategy to enhance bioavailability and anticancer efficacy of curcumin. PLoS One 9, article no. e89351.
Schacterle, G and Pollack, R (1973) Simplified method for quantitative assay of small amounts of protein in biological material. Analytical Biochemistry 51, 654655.
Sharma, S, Kumar, M, Sharma, S, Nargotra, A, Koul, S and Khan, IA (2010) Piperine as an inhibitor of Rv1258c, a putative multidrug efflux pump of Mycobacterium tuberculosis. Journal of Antimicrobial Chemotherapy 65, 16941701.
Srinivasan, K (2007) Black pepper and its pungent principle-piperine: a review of diverse physiological effects. Critical Reviews in Food Science and Nutrition 47, 735748.
Srinivasan, K (2009) Black pepper (Piper nigrum) and its bioactive compound piperine. In Aggarwal, BB and Kunnumakkara, AB (eds), Molecular Targets and Therapeutic Uses of Spices: Modern Uses for Ancient Medicine. Singapore: World Scientific Publishing Company, pp. 2564.
Stamets, PE, Naeger, NL, Evans, JD, Han, JO, Hopkins, BK, Lopez, D, Moershel, HM, Nally, R, Sumerlin, D, Taylor, AW, Carris, LM and Sheppard, WS (2018) Extracts of polypore mushroom mycelia reduce viruses in honey bees. Scientific Reports 8, article no. 13936.
Strachecka, A, Gryzinska, M and Krauze, M (2010) The influence of environmental pollution on the protective proteolytic barrier of the honeybee Apis mellifera mellifera. Polish Journal of Environmental Studies 19, 855859.
Strachecka, A, Krauze, M, Olszewski, K, Borsuk, G, Paleolog, J, Merska, M, Chobotow, J, Bajda, M and Grzywnowicz, K (2014 a) Unexpectedly strong effect of caffeine on the vitality of western honeybees (Apis mellifera). Biochemistry – Moscow 79, 11921201.
Strachecka, A, Olszewski, K, Paleolog, J, Borsuk, G, Bajda, M, Krauze, M, Merska, M and Chobotow, J (2014 b) Coenzyme Q10 treatments influence the lifespan and key biochemical resistance systems in the honeybee, Apis mellifera. Archives of Insect Biochemistry and Physiology 86, 165179.
Strachecka, A, Olszewski, K and Paleolog, J (2015) Curcumin stimulates biochemical mechanisms of Apis mellifera resistance and extends the apian life-span. Journal of Apicultural Science 59, 129141.
Strachecka, A, Olszewski, K and Paleolog, J (2016) Varroa treatment with bromfenvinphos markedly suppresses honeybee biochemical defence levels. Entomologia Experimentalis et Applicata 160, 5771.
Suwannapong, G, Maksong, S, Phainchajoen, M, Benbow, ME and Mayack, C (2018) Survival and health improvement of Nosema infected Apis florea (Hymenoptera: Apidae) bees after treatment with propolis extract. Journal of Asia-Pacific Entomology 21, 437444.
Tasleem, F, Azhar, I, Ali, SN, Perveen, S and Mahmood, ZA (2014) Analgesic and anti-inflammatory activities of Piper nigrum L. Asian Pacific Journal of Tropical Medicine 7S1, S461S468.
Tolfsen, CC, Baker, N, Kreibich, C and Amdam, GV (2011) Flight restriction prevents associative learning deficits but not changes in brain protein-adduct formation during honeybee ageing. Journal of Experimental Biology 214, 13221332.
Umar, S, Sarwar, AHMG, Umar, K, Ahmad, N, Sajad, M, Ahmad, S, Katiyar, CK and Khan, HA (2013) Piperine ameliorates oxidative stress, inflammation and histological outcome in collagen induced arthritis. Cellular Immunology 284, 5159.
vanEngelsdorp, D, Traynor, KS, Andree, M, Lichtenberg, EM, Chen, Y, Saegerman, C and Cox-Foster, DL (2017) Colony Collapse Disorder (CCD) and bee age impact honey bee pathophysiology. PLoS ONE 12, article no. e0179535.
Vannette, RL, Mohamed, A and Johnon, BR (2015) Forager bees (Apis mellifera) highly express immune and detoxification genes in tissues associated with nectar processing. Scientific Reports 5, article no. 16224.
Vasavirama, K and Upender, M (2014) Piperine: a valuable alkaloid from piper species. International Journal of Pharmacy and Pharmaceutical Sciences 6, 3438.
Warholm, M, Guthenberg, C, von Bahr, C and Mannervik, B (1985) Glutathione transferases from human liver. Methods in Enzymology 113, 499504.
Williams, JB, Roberts, SP and Elekonich, MM (2008) Age and natural metabolically-intensive behavior affect oxidative stress and antioxidant mechanisms. Experimental Gerontology 43, 538549.
Zou, Z, Lopez, DL, Kanost, MR, Evans, JD and Jiang, H (2006) Comparative analysis of serine protease-related genes in the honey bee genome: possible involvement in embryonic development and innate immunity. Insect Molecular Biology 15, 603614.


Piperine as a new natural supplement with beneficial effects on the life-span and defence system of honeybees

  • M. Schulz (a1), A. Łoś (a1) (a2), M. Grzybek (a3) (a4), R. Ścibior (a3) and A. Strachecka (a1)...


Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed