DNA extraction and PCR detection of Paenibacillus larvae spores from naturally contaminated honey and bees using spore-decoating and freeze-thawing techniques

World Journal of Microbiology and Biotechnology

Volumn 23, Issue 4, 2007, Pages 593-597

  D'Alessandro, Bruno ^a   Antúnez, Karina ^a   Piccini, Claudia ^a   Zunino, Pablo ^a  

^a INSTITUTO DE INVESTIGACIONES BIOLÓGICAS CLEMENTE ESTABLE   (Uruguay)

Author keywords

American foulbrood; Freeze thawing; Paenibacillus larvae; Polymerase chain reaction; Spore decoating; Spores

Indexed keywords

BACTERIA; BIOASSAY; CONTAMINATION; DNA; FREEZING; THAWING;

AMERICAN FOULBROOD; FREEZE-THAWING; PAENIBACILLUS LARVAE; POLYMERASE CHAIN REACTION; SPORE-DECOATING;

DISEASES;

APIS MELLIFERA; APOIDEA; BACTERIA (MICROORGANISMS); PAENIBACILLUS; PAENIBACILLUS LARVAE;

EID: 33947545559     PISSN: 09593993     EISSN: None     Source Type: Journal    
DOI: 10.1007/s11274-006-9261-y     Document Type: Article

References (14)

1. Alippi AM, López AC, Aguilar OM (2002) Differentiation of Paenibacillus larvae subsp. larvae, the cause of American foulbrood of honeybees, by using PCR and restriction fragment analysis of genes encoding 16S rRNA. Appl Environ Microbiol 68: 3655-3660
2. Bickley J, Owen R (1995) Preparation of bacterial genomic DNA. In: Howard J, Whitcombe DM (eds) Diagnostic bacteriology protocols. Humana Press, Totowa, New Jersey, pp 141-147 (ISBN 0896032973)
3. Govan VA, Allsopp MH, Davison S (1999) A PCR detection method for rapid identification of Paenibacillus larvae. Appl Environ Microbiol 65:2243-2245
4. Hornitzky MAZ, Nicholls PJ (1993) J medium is superior to sheep blood agar and brain infusion agar for the isolation of Bacillus larvae from honey samples. J Apic Res 32:51-52
5. Lauro FM, Favaretto M, Covolo L, Rassu M, Bertoloni G (2003) Rapid detection of Paenibacillus larvae from honey and hive samples with a novel nested PCR protocol. Int J Food Microbiol 81:195-201
6. Levi K, Higham JL, Coates D, Hamlyn PF (2003) Molecular detection of anthrax spores on animal fibres. Lett Appl Microbiol 36:418-422
7. Paidhungat M, Setlow P (2000) Role of Ger proteins in nutrient and nonnutrient triggering of spore germination in Bacillus subtilis. J Bacteriol 182:2513-2519
8. 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 Microbiol Biotechnol 18: 761-765
9. Riesenman PJ, Nicholson WL (2000) Role of the spore coat layers in Bacillus subtilis spore resistance to hydrogen peroxide, artificial UV-C, UV-B, and solar UV radiation. Appl Environ Microbiol 66:620-626
10. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
11. Sargent MG (1980) A procedure for isolating high quality DNA from spores of Bacillus subtilis 168. J Gen Microbiol 116:511-514
12. Setlow B, Setlow P (1998) Heat killing of Bacillus subtilis spores in water is not due to oxidative damage. Appl Environ Microbiol 64:4109-4112
13. Vary JC (1973) Germination of Bacillus megaterium spores after various extraction procedures. J Bacteriol 116:797-802
14. Wilson IG (1997) Inhibition and facilitation of nucleic acid amplification. Appl Environ Microbiol 63:3741-3751