Short communication: Natural molecules for the control of Paenibacillus larvae, causal agent of American foulbrood in honey bees (Apis mellifera L.)

  • Pablo Giménez-Martínez Universidad Nacional de Mar del Plata, Centro de Investigación en Abejas Sociales (CIAS-IIPROSAM), Funes 3350, 7600 Mar del Plata, Argentina. Agencia Nacional de Promoción Científica y Tecnológica, Godoy Cruz 2370, C1425FQD, Buenos Aires,
  • Noelia Cugnata Universidad Nacional de Mar del Plata, Centro de Investigación en Abejas Sociales (CIAS-IIPROSAM), Funes 3350, 7600 Mar del Plata
  • Rosa M. Alonso-Salces Universidad Nacional de Mar del Plata, Centro de Investigación en Abejas Sociales (CIAS-IIPROSAM), Funes 3350, 7600 Mar del Plata Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2290, C1425FQD Buenos Aires
  • Daniela Arredondo Instituto de Investigaciones Biológicas Clemente Estable, Dept. Microbiología, Av. Italia 3318, 11600. Montevideo
  • Karina Antunez Instituto de Investigaciones Biológicas Clemente Estable, Dept. Microbiología, Av. Italia 3318, 11600. Montevideo
  • Rosana De Castro Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Godoy Cruz 2290, C1425FQD Buenos Aires Universidad Nacional de Mar del Plata, Instituto de Investigaciones Biológicas (IIB) Funes 3350, 7600 Mar del Plata
  • Sandra R. Fuselli Universidad Nacional de Mar del Plata, Centro de Investigación en Abejas Sociales (CIAS-IIPROSAM), Funes 3350, 7600 Mar del Plata Comisión Investigaciones Científicas de la Provincia de Buenos Aires (CIC), Calle 526 entre 10 y 11, B1900, La Plata
Keywords: antimicrobial activity, metalloproteases

Abstract

Aim of study: To evaluate the potential bactericidal activity of natural molecules against Paenibacillus larvae. Moreover, we investigated if molecules that exhibit antimicrobial activity were able to inhibit the proteolytic activity of the bacterium.

Area of study: Isolates S1 and S2 were from Balcarce, Buenos Aires province, strain S3 from Rio Cuarto, Cordoba province, strain S4 from Concordia, Entre Rios province, strain S5 and S8 from Necochea, Buenos Aires, strain S6 and S7 from Mar del Plata, Buenos Aires, strain S9 from Modena, Italy and strain S10 from Emilia Reggio, Italy.

Material and methods: Bacterial isolates identification was carried out by amplification of a specific 16S rRNA gene fragment of P. larvae using primers PL5 and PL4. Screening of the antimicrobial activity of thirteen molecules against four P. larvae isolates was conducted by the agar diffusion technique. The antimicrobial activity of selected molecules was evaluated by broth microdilution method.

Main results: Menadione, lauric acid, monoglyceride of lauric acid and naringenin showed antimicrobial activity against ten P. larvae isolates. Menadione and lauric acid showed the strongest activities, with minimum inhibitory concentration mean values ranging 0.78-3.125 µg/mLand 25-50 µg/mL, respectively.

Research highlights: Those concentrations are feasible to be applied at field level, and constitute promissory candidates to be evaluated using in vivo larval models.

Downloads

Download data is not yet available.

Author Biography

Sandra R. Fuselli, Universidad Nacional de Mar del Plata, Centro de Investigación en Abejas Sociales (CIAS-IIPROSAM), Funes 3350, 7600 Mar del Plata Comisión Investigaciones Científicas de la Provincia de Buenos Aires (CIC), Calle 526 entre 10 y 11, B1900, La Plata

 

 

References

Alippi AM, Ringuelet JA, Cerimele EL, Re MS, Henning CP, 1996. Antimicrobial activity of some essential oils against Paenibacillus larvae, the causal agent of American Foulbrood Disease. J Herb Spic Med Plants 4: 9-16. https://doi.org/10.1300/J044v04n02_03

Alippi AM, Aguilar OM, 1998. Unique DNA fingerprint patterns of Paenibacillus larvae subsp. larvae strains. J Apic Res 37: 273-280. https://doi.org/10.1080/00218839.1998.11100983

Alippi AM, Reynaldi FJ, López AC, De Giusti MR, Aguilar OM, 2004. Molecular epidemiology of Paenibacillus larvae larvae and incidence of American foulbrood in Argentinean honeys from Buenos Aires province. J Apic Res 43: 135-143. https://doi.org/10.1080/00218839.2004.11101124

Ansari MJ, Al-Ghamdi A, Usmani S, Al-Waili N, Nuru A, Sharma D, Khan KA, Kaur M, Omer M, 2016. In vitro evaluation of the effects of some plant essential oils on Paenibacillus larvae, the causative agent of American foulbrood. Biotechnol Biotechnol Equip 30: 49-55. https://doi.org/10.1080/13102818.2015.1086690

Antúnez K, Piccini C, Castro-Sowinski S, Rosado AS, Seldin L, Zunino P, 2007. Phenotypic and genotypic characterization of Paenibacillus larvae isolates. Vet Microbiol 124: 178-183. https://doi.org/10.1016/j.vetmic.2007.04.012

Antúnez K, Harriet J, Gende L, Maggi M, Eguaras M, Zunino P, 2008. Efficacy of natural propolis extract in the control of American Foulbrood. Vet Microbiol 131: 324-331. https://doi.org/10.1016/j.vetmic.2008.04.011

Antúnez K, Martín-Hernández R, Prieto L, Meana A, Zunino P, Higes M, 2009. Immune suppression in the honey bee (Apis mellifera) following infection by Nosema ceranae (Microsporidia). Environ Microbiol 11: 2284-2290. https://doi.org/10.1111/j.1462-2920.2009.01953.x

Bíliková K, Popova M, Trusheva B, Bankova V, 2013. New anti-Paenibacillus larvae substances purified from propolis. Apidologie 44: 278-285. https://doi.org/10.1007/s13592-012-0178-1

Boligon AA, de Brum TF, Zadra M, Piana M, dos Santos ACF, Fausto VP, Júnior VdSB, de Almeida Vaucher R, Santos RCV, Athayde ML, 2013. Antimicrobial activity of Scutia buxifolia against the honeybee pathogen Paenibacillus larvae. J Invertebr Pathol 112: 105-107. https://doi.org/10.1016/j.jip.2012.11.009

Bonev B, Hooper J, Parisot J, 2008. Principles of assessing bacterial susceptibility to antibiotics using the agar diffusion method. J Antimicr Chemother 61: 1295-1301. https://doi.org/10.1093/jac/dkn090

Chaimanee V, Thongtue U, Sornmai N, Songsri S, Pettis JS, 2017, Antimicrobial activity of plant extracts against the honeybee pathogens, Paenibacillus larvae and Ascosphaera apis and their topical toxicity to Apis mellifera adults. J App Microbiol 123: 1160-1167. https://doi.org/10.1111/jam.13579

Cugnata N, Guaspari E, Pellegrini M, Fuselli S, Alonso-Salces R, 2017. Optimal concentration of organic solvents to be used in the broth microdilution method to determine the antimicrobial activity of natural products against Paenibacillus larvae. J Apic Sci: 61:37-53 https://doi.org/10.1515/jas-2017-0004

Damiani N, Fernández NJ, Porrini MP, Gende LB, Álvarez E, Buffa F, Brasesco C, Maggi MD, Marcangeli JA, Eguaras MJ, 2014. Laurel leaf extracts for honeybee pest and disease management: Antimicrobial, microsporicidal, and acaricidal activity. Parasitol Res 113: 701-709. https://doi.org/10.1007/s00436-013-3698-3

Dancer BN, Chantawannakul P, 1997. The proteases of American foulbrood scales. J Invertebr Pathol 70: 79-87. https://doi.org/10.1006/jipa.1997.4672

Dingman DW, Stahly DP, 1983. Medium promoting sporulation of Bacillus larvae and metabolism of medium components. Appl Environ Microbiol 46: 860-869.

Djukic M, Brzuszkiewicz E, Fünfhaus A, Voss J, Gollnow K, Poppinga L, Liesegang H, Garcia-Gonzalez E, Genersch E, Daniel R, 2014. How to kill the honey bee larva: genomic potential and virulence mechanisms of Paenibacillus larvae. PLoS ONE 9: e90914. https://doi.org/10.1371/journal.pone.0090914

Duarte MCT, Leme EE, Delarmelina C, Soares AA, Figueira GM, Sartoratto AJJ, 2007 Activity of essential oils from Brazilian medicinal plants on Escherichia coli. J Ethnopharmacol 111: 197-201. https://doi.org/10.1016/j.jep.2006.11.034

Feldlaufer M, Lusby WR, Knox D, Shimanuki H, 1993. Isolation and identification of linoleic acid as an antimicrobial agent from the chalkbrood fungus, Ascosphaera apis. Apidologie 24: 89-94. https://doi.org/10.1051/apido:19930201

Flesar J, Havlik J, Kloucek P, Rada V, Titera D, Bednar M, Stropnicky M, Kokoska L, 2010. In vitro growth-inhibitory effect of plant-derived extracts and compounds against Paenibacillus larvae and their acute oral toxicity to adult honey bees. Vet Microbiol 145: 129-133. https://doi.org/10.1016/j.vetmic.2010.03.018

Fuselli SR, De La Rosa SBG, Gende LB, Eguaras MJ, Fritz R, 2006. Antimicrobial activity of some Argentinian wild plant essential oils against Paenibacillus larvae larvae, causal agent of American foulbrood (AFB). J Apic Res 45: 2-7. https://doi.org/10.1080/00218839.2006.11101304

Genersch E, 2010. American Foulbrood in honeybees and its causative agent, Paenibacillus larvae. J Invertebr 103: 10-19. https://doi.org/10.1016/j.jip.2009.06.015

Genersch E, Otten C, 2003. The use of repetitive element PCR fingerprinting (rep-PCR) for genetic subtyping of German field isolates of Paenibacillus larvae subsp. larvae. Apidologie 34: 195-206. https://doi.org/10.1051/apido:2003025

Genersch E, Forsgren E, Pentikäinen J, Ashiralieva A, Rauch S, Kilwinski J, Fries I, 2006. Reclassification of Paenibacillus larvae subsp. pulvifaciens and Paenibacillus larvae subsp. larvae as Paenibacillus larvae without subspecies differentiation. Int J Sys Evol Microbiol 56: 501-511. https://doi.org/10.1099/ijs.0.63928-0

Hansen H, Brødsgaard CJ, 1999. American foulbrood: A review of its biology, diagnosis and control. Bee World 80: 5-23. https://doi.org/10.1080/0005772X.1999.11099415

Hornitzky MAZ, Nicholls PJ, 1993. J medium is superior to sheep blood agar and brain heart infusion agar for the isolation of Bacillus larvae from honey samples. J Apic Res 32: 51-52. https://doi.org/10.1080/00218839.1993.11101287

Hornitzky MAZ, 2003. Fatty acids: An alternative control strategy for honeybee diseases. Rural Ind Res & Dev Corp, Barton, A.C.T., Australia.

Isidorov VA, Buczek K, Zambrowski G, Miastkowski K, Swiecicka I, 2017. In vitro study of the antimicrobial activity of European propolis against Paenibacillus larvae. Apidologie 48: 411-422. https://doi.org/10.1007/s13592-016-0485-z

Laemmli UK, 1970. Cleavage of structural proteins during the assembly of the head of Bacteriophage T4. Nature 227: 680-685. https://doi.org/10.1038/227680a0

Lokvam J, Braddock JF, Reichardt PB, Clausen TP, 2000. Two polyisoprenylated benzophenones from the trunk latex of Clusia grandiflora (Clusiaceae). Phytochemistry 55: 29-34. https://doi.org/10.1016/S0031-9422(00)00193-X

Martel AC, Zeggane S, Drajnudel P, Faucon JP, Aubert M, 2006. Tetracycline residues in honey after hive treatment. Food Addit Contam 23: 265-273. https://doi.org/10.1080/02652030500469048

Michielin EM, Salvador AA, Riehl CA, Smânia Jr A, Smânia EF, Ferreira SR, 2009. Chemical composition and antibacterial activity of Cordia verbenacea extracts obtained by different methods. Bioresour Technol 100: 6615-6623. https://doi.org/10.1016/j.biortech.2009.07.061

Miyagi T, Peng CYS, Chuang RY, Mussen EC, Spivak MS, Doi RH, 2000. Verification of oxytetracycline-resistant American foulbrood pathogen Paenibacillus larvae in the United States. J Invertebr 75: 95-96. https://doi.org/10.1006/jipa.1999.4888

Nordström S, Fries I, 1995. A comparison of media and cultural conditions for identification of Bacillus larvae in honey. J Apic Res 34: 97-103. https://doi.org/10.1080/00218839.1995.11100894

Piccini C, D'Alessandro B, Antúnez K, Zunino P, 2002. Detection of Paenibacillus larvae subspecies larvae spores in naturally infected bee larvae and artificially contaminated honey by PCR. World J Microb Biot 18: 761-765.

Sabaté DC, Gonzaléz MJ, Porrini MP, Eguaras MJ, Audisio MC, Marioli JM, 2012. Synergistic effect of surfactin from Bacillus subtilis C4 and Achyrocline satureioides extracts on the viability of Paenibacillus larvae. World J Microb Biot 28: 1415-1422. https://doi.org/10.1007/s11274-011-0941-x

Shimanuki H, Knox DA, Feldlaufer MF, 1992. Honey bee disease interactions: The impact of chalkbrood on other honey bee brood diseases. Am Bee J 132: 735-736.

Tutun H, Koç N, Kart A, 2018. Plant essential oils used against some bee diseases. TURJAF 6: 34-45. https://doi.org/10.24925/turjaf.v6i1.34-45.1502

Versalovic J, Schneider M, De Bruijn FJ, Lupski JR, 1994. Genomic fingerprinting of bacteria using repetitive sequence-based polymerase chain reaction. Met Mol Cel Bio 5: 25-40.

Wang YS, He HP, Yang JH, Di YT, Hao XJJM, 2008. New monoterpenoid coumarins from Clausena anisum-olens. Molecules 13: 931-937. https://doi.org/10.3390/molecules13040931

Published
2019-11-08
How to Cite
Giménez-Martínez, P., Cugnata, N., Alonso-Salces, R. M., Arredondo, D., Antunez, K., De Castro, R., & Fuselli, S. R. (2019). Short communication: Natural molecules for the control of Paenibacillus larvae, causal agent of American foulbrood in honey bees (Apis mellifera L.). Spanish Journal of Agricultural Research, 17(3), e05SC01. https://doi.org/10.5424/sjar/2019173-14740
Section
Animal health and welfare