Evaluation of the bacterial diversity of Pressure ulcers using bTEFAP pyrosequencing

Abstract

BackgroundDecubitus ulcers, also known as bedsores or pressure ulcers, affect millions of hospitalized patients each year. The microflora of chronic wounds such as ulcers most commonly exist in the biofilm phenotype and have been known to significantly impair normal healing trajectories.MethodsBacterial tag-encoded FLX amplicon pyrosequencing (bTEFAP), a universal bacterial identification method, was used to identify bacterial populations in 49 decubitus ulcers. Diversity estimators were utilized and wound community compositions analyzed in relation to metadata such as Age, race, gender, and comorbidities.ResultsDecubitus ulcers are shown to be polymicrobial in nature with no single bacterium exclusively colonizing the wounds. The microbial community among such ulcers is highly variable. While there are between 3 and 10 primary populations in each wound there can be hundreds of different species present many of which are in trace amounts. There is no clearly significant differences in the microbial ecology of decubitus ulcer in relation to metadata except when considering diabetes. The microbial populations and composition in the decubitus ulcers of diabetics may be significantly different from the communities in non-diabetics.ConclusionsBased upon the continued elucidation of chronic wound bioburdens as polymicrobial infections, it is recommended that, in addition to traditional biofilm-based wound care strategies, an antimicrobial/antibiofilm treatment program can be tailored to each patient's respective wound microflora.

Antibiofilm efficacy of honey and bee-derived defensin-1 on multispecies wound biofilm

“…The tissue of all chronic wounds is colonized by polymicrobial flora (Smith et al, 2010). Over 90% of chronic wounds contain bacteria and fungi from the skin, oral mucosa, enteric tract or environment.…”

Influence of Microorganisms on the Healing of Skin Grafts from Chronic Venous Leg Wounds

“…It differs from the classic Sanger sequencing, in that it relies on the detection of pyrophosphate release upon nucleotide incorporation, rather than chain termination with dideoxynucleotides (Ronaghi et al, 1998). This deep sequencing technique has been used to evaluate the diversity of microbial populations where thousands of reads/sequences can be www.intechopen.com Skin Grafts -Indications, Applications and Current Research 280 obtained from each sample (Andersson et al, 2007;Bogaert et al;Dowd et al, 2008;Price et al, 2010;Smith et al, 2010;Stewart et al, 2010;Wolcott et al, 2009). …”

“…In a retrospective analysis of the DNA resulting from the wounds included in the study, qPCR with universal fungal primers and probe revealed fungal DNA in one of 14 samples. In a recent study by Smith (2010) pyrosequencing was used to identify bacterial populations in 49 pressure ulcers. Diversity estimators were utilized and wound community compositions analyzed in relation to metadata such as age, race, gender, and comorbidities.…”

“…Adding detection of known virulence genes to the molecular methods would be helpful in the process of interpretation. Ideally, each wound should be carefully evaluated and in addition to traditional biofilmbased wound care strategies, an antimicrobial/antibiofilm treatment program with individualized therapeutic approaches should be identified for each patient's respective wound microflora (Smith et al, 2010;Wolcott et al, 2009). However, we do not have the tools for interpretation yet.…”

Antibiotic Resistance in Nursing Homes

“…Culture results must be interpreted with caution, because should not be used as the sole criterion for infections, without clinical or histopathological evidence of infection (Hirshberg J & Al, 2000). Despite the aforementioned information, there is agreement on the most frequently isolated bacteria, including Staphylococcus aureus, betaHemolytic Streptococci, Gram negative organisms (including Enterobacteriaceae and Pseudomonas spp, and other Gram positive organisms such Enterococcus spp) and Anaerobic organisms (Chow AW & Al, 1977;Sapico FI & Al, 1986;Muder RR & Al, 1992;Nicolle LE & Al, 1994b;Smith DM & Al, 2010;Lund-Nielsen B & Al, 2011 …”