The Burkholderia cepacia complex (BCC) includes opportunistic pathogenic bacteria that have occasionally been recovered from various pharmaceutical products, including antiseptics and disinfectants. Plausible reasons for the contamination include intrinsic sources, such as inadequate process controls, especially for water or equipment used during product manufacture, or extrinsic sources, such as improper handling and dilution or distribution in contaminated containers. Because the survival of BCC in antiseptics is a concern to the public health and pharmaceutical industry, we determined minimum inhibitory concentrations (MICs) of 36 BCC strains against the antiseptics, following exposure to chlorhexidine gluconate (CHX) and benzalkonium chloride (BZK) solutions (1-500 µg/ml for each chemical). Susceptibility to CHX and BZK varied across the BCC strains and was recorded as mean 90.3 and 111.1 µg/ml, respectively, at initial inoculation, which was significantly higher than the 46.4 and 61.1 µg/ml levels measured for BCC incubated in water for 40 days. After determining antiseptic MICs of individual BCC strains, BCC recovery was measured on Tryptic Soy Agar (TSA), Reasoner's Second Agar (R2A) and diluted preparations of these media under their sub-MICs. The survival of BCC was monitored for 14 days (336 h) in sub-MICs diluted to less than their antiseptic susceptible concentration value. Diluted TSA and R2A media exhibited greater efficiency of recovery for most BCC strains from the CHX and BZK solutions than full strength TSA or R2A. For BCC survival in antiseptic solutions, the cell number of BCC decreased rapidly within the first 20 min in both antiseptics, but after this, recovery remained constant in CHX and increased in BZK over the 14 day incubation period. The results indicate that BCC in water can remain viable with low susceptibility to antiseptics for 14 days, which suggests the necessity for improved detection methods and control measures to monitor BCC contamination in pharmaceutical products.

A reductively dehalogenating, strictly anaerobic, sulfate-reducing bacterium, designated strain AA1 T , was isolated from the marine sponge Aplysina aerophoba collected in the Mediterranean Sea and was characterized phenotypically and phylogenetically. Cells of strain AA1 T were Gramnegative, short, curved rods. Growth of strain AA1 T was observed between 20 and 37 6C (optimally at 28 6C) at pH 7-8. NaCl was required for growth; optimum growth occurred in the presence of 25 g NaCl l -1 . Growth occurred with lactate, propionate, pyruvate, succinate, benzoate, glucose and sodium citrate as electron donors and carbon sources and either sulfate or 2-bromophenol as electron acceptors, but not with acetate or butyrate. Strain AA1 T was able to dehalogenate several different bromophenols, and 2-and 3-iodophenol, but not monochlorinated or fluorinated phenols. Lactate, pyruvate, fumarate and malate were not utilized without an electron acceptor. The G+C content of the genomic DNA was 58.5 mol%. The predominant cellular fatty acids were C 14 : 0 , iso-C 14 : 0 , C 14 : 0 3-OH, anteiso-C 15 : 0 , C 16 : 0 , C 16 : 1 v7c and C 18 : 1 v7c. Phylogenetic analysis based on 16S rRNA gene sequence comparisons placed the novel strain within the class Deltaproteobacteria. Strain AA1 T was related most closely to the type strains of Desulfoluna butyratoxydans (96 % 16S rRNA gene sequence similarity), Desulfofrigus oceanense (95 %) and Desulfofrigus fragile (95 %). Based on its phenotypic, physiological and phylogenetic characteristics, strain AA1 T is considered to represent a novel species of the genus Desulfoluna, for which the name Desulfoluna spongiiphila sp. nov. is proposed. The type strain is AA1 T (5DSM 17682 T 5ATCC BAA-1256 T ).The marine environment is a particularly rich source of biogenic organohalides, which are produced by a diversity of marine organisms, including molluscs, algae, polychaetes, jellyfish and sponges (Ashworth & Cormier, 1967;Baker & Duke, 1973;Fielman et al., 1999; Garson et al., 1994;Schmitz & Gopichand, 1978;White & Hager, 1977).A number of sponges (phylum Porifera), such as species of the genus Aplysina, have been shown to produce a wide variety of brominated metabolites, including bromoindoles, bromophenols, polybrominated diphenyl ethers and brominated dibenzo-p-dioxins (Ebel et al., 1997;Gribble, 1999; Norte & Fernández, 1987;Utkina et al., 2001). Brominecontaining metabolites can account for over 10 % of the sponge dry weight (Turon et al., 2000). These compounds may serve as a chemical defence against predators and may inhibit biofouling (Weiss et al., 1996). In addition, Aplysina sponges harbour large amounts of bacteria, which can amount to 40 % of the biomass of the animal, and it has been hypothesized that some of the organobromine compounds may in fact be synthesized by bacteria associated with the sponge (Hentschel et al., 2001(Hentschel et al., , 2003.As the marine environment is a particularly rich source of biogenic organohalides, it is not surprising that dehalogenating bacteria ...

Pharmaceutical products that are contaminated with Burkholderia cepacia complex (BCC) bacteria may pose serious consequences to vulnerable patients. Benzyldimethylalkylammonium chloride (BZK) cationic surfactants are extensively used in medical applications and have been implicated in the coselection of antimicrobial resistance. The ability of BCC to degrade BZK, tetradecyldimethylbenzylammonium chloride (C14BDMA-Cl), dodecyldimethylbenzylammonium chloride (C12BDMA-Cl), decyldimethylbenzylammonium chloride (C10BDMA-Cl), hexyldimethylbenzylammonium chloride, and benzyltrimethylammonium chloride was determined by incubation in 1/10-diluted tryptic soy broth (TSB) to determine if BCC bacteria have the ability to survive and inactivate these disinfectants. With BZK, C14BDMA-Cl, and C12BDMA-Cl, inhibition of the growth of 20 BCC strains was observed in disinfectant solutions that ranged from 64 to 256 µg/ml. The efflux pump inhibitor carbonyl cyanide m-chlorophenylhydrazone increased the sensitivity of bacteria to 64 µg/ml BZK. The 20 BCC strains grew well in 1/10-diluted TSB medium with BZK, C12BDMA-Cl, and C10BDMA-Cl; they absorbed and degraded the compounds in 7 days. Formation of benzyldimethylamine and benzylmethylamine as the initial metabolites suggested that the cleavage of the C alkyl-N bond occurred as the first step of BZK degradation by BCC bacteria. Proteomic data confirmed the observed efflux activity and metabolic inactivation via biodegradation in terms of BZK resistance of BCC bacteria, which suggests that the two main resistance mechanisms are intrinsic and widespread.