Genetic Diversity of Burkholderia contaminans Isolates from Cystic Fibrosis Patients in Argentina

Abstract

A total of 120 Burkholderia cepacia complex isolates collected during 2004 -2010 from 66 patients in two cystic fibrosis reference centers in Argentina were analyzed. Burkholderia contaminans was the species most frequently recovered (57.6%), followed by Burkholderia cenocepacia (15%), a species distribution not reported so far. The recA-PCR-based techniques applied to the B. contaminans isolates revealed that 85% of the population carried the recA-ST-71 allele. Our results showed the utility of BOX-PCR genotyping in analyzing B. contaminans diversity. This approach allowed us to address clonal transmission during an outbreak and the genetic changes occurring in infecting bacteria over the course of chronic infection. Burkholderia cepacia complex species are capable of causing chronic and often severe respiratory tract infections in cystic fibrosis (CF) patients and other types of infections in immunocompromised patients. Although many CF patients remain infected by these bacteria and yet stay relatively healthy for prolonged periods, others either have a severe decline in their pulmonary status or die shortly after the initial colonization (1). In Argentina, the first reports of B. cepacia complex bacteria infecting CF patients started around 1990. At that time, Burkholderia spp. were recovered sporadically and with a very low prevalence (Ͻ0.1%). The prevalence increased over the last decade from 0.2 to 3.6%, depending on the medical center. In early 2004, an outbreak involving main Argentine CF care centers occurred, and the proportion of patients from whom B. cepacia complex species were recovered ranged from 19 to 36% (2-4). Currently, through strict infection control procedures, the prevalence of Burkholderia spp. in local patients has decreased to approximately 10%. Recent worldwide surveillance studies concerning the distribution of B. cepacia complex species in CF patients have cited B. multivorans and B. cenocepacia as the most frequently recovered species. These species account for approximately 80% of the infected patients, and the prevalence of one or the other is geographically and temporally dependent (5-7). Unlike this worldwide situation, in Argentina a relatively high prevalence of B. contaminans followed by B. cenocepacia has been observed (3, 4). To our knowledge, no other geographical region has been reported in the literature to have such a high occurrence of B. contaminans infecting CF patients. In view of this particular scenario, coupled with the lack of biodiversity information available for B. contaminans species, we decided to study this local population by recA-PCR-based techniques and by repetitive element sequence-based PCR (rep-PCR) approaches. These methods allowed us to address the genetic diversity of B. contaminans isolates recovered during that outbreak along with the genetic changes occurring in the infecting bacteria over the course of a chronic infection.

SNaPBceBcon : a Practical Tool for Identification and Genotyping of Burkholderia cepacia and Burkholderia contaminans

“…In addition, the presence of the same SNaP profile in consecutive samples from individual patients was common in patients colonized by B. cepacia or B. contaminans, as well as the isolation of clonal variants (SNaP profiles with one or two differences) in the same patients, presumably resulting from adaptive evolution in the CF lung during chronic infection (26). Results similar to these shown by the SNaPBceBcon assay were previously reported by MLST (16,17,19,21,25).…”

“…B. contaminans has been isolated worldwide from both environmental and clinical samples (15)(16)(17)(18)(19)(20)(21). B. contaminans has a low prevalence in CF patients worldwide, with remarkable exceptions registered in Argentina (21), Brazil (15,22), Spain (23), and Portugal (17).…”

Incidence of Burkholderia contaminans at a cystic fibrosis centre with an unusually high representation of Burkholderia cepacia during 15 years of epidemiological surveillance

“…in seven patients -three girls and four boys -of a total of 41 patients). The prevalence of B. contaminans in CF patients worldwide is low, with remarkable exceptions registered in Argentina (Martina et al, 2013) and Spain (Medina-Pascual et al, 2015), presumably resulting from patient to patient transmission or other sources of infection, such as environmental or pharmaceutical products.…”

“…The group of previously unclassified Bcc isolates of taxon K (Vermis et al, 2002;Baldwin et al, 2005;Dalmastri et al, 2005Dalmastri et al, , 2007Payne et al, 2005;Mahenthiralingam et al, 2006) includes bacteria isolated worldwide from human and environmental sources, namely from sputum cultures of CF patients in the UK, Italy, Portugal, USA, Canada, China, Brazil, Argentina and Australia (Cunha et al, 2003(Cunha et al, , 2007Campana et al, 2005;Assaad et al, 2006;Mahenthiralingam et al, 2006;Jordá-Vargas et al, 2008;Fang et al, 2010;Martina et al, 2013;Ramsay et al, 2013), as well as from river water, human-built water reservoirs, soil, roots, animals, pharmaceutical products, such as contaminated nasal spray, personal care products and domestic products (Souza et al, 2004;Mahenthiralingam et al, 2006Mahenthiralingam et al, , 2008Martin et al, 2011). B. contaminans has a low prevalence in CF patients worldwide, with remarkable exceptions in Argentina and in Spain (Martina et al, 2013). Several reports of B. contaminans outbreaks amongst non-CF patients in Argentina and Brazil have also been also published, suggesting the existence of regional spreading of B. contaminans and other Bcc species between bordering countries (Mahenthiralingam et al, 2000;Agodi et al, 2002;Magalhães et al, 2003;Shehabi et al, 2004;Souza et al, 2004;Woods et al, 2004).…”

The Tyrosine Kinase BceF and the Phosphotyrosine Phosphatase BceD of Burkholderia contaminans Are Required for Efficient Invasion and Epithelial Disruption of a Cystic Fibrosis Lung Epithelial Cell Line

“…Depending on the B. cepacia complex strain, this colonization ranges from asymptomatic to a rapid decline of lung function characterized by a necrotizing pneumonia and the development of septicemia known as cepacia syndrome (19). Burkholderia cenocepacia and Burkholderia multivorans are the predominant species in infected CF patients (20,21), but an increasing number of outbreaks caused by B. cepacia complex species such as Burkholderia contaminans has been reported (22)(23)(24). B. cepacia complex species are also intrinsically resistant to several antibiotics and able to form biofilms, making their eradication from both lungs and clinical devices very difficult (25)(26)(27).…”