Isolation of a Rochalimaea-like organism from a febrile patient infected with human immunodeficiency virus was confirmed. Analysis of 16S rRNA gene sequences, together with polymerase chain reaction and restriction endonuclease length polymorphism analysis of a portion of the citrate synthase gene, demonstrated that the agent is closely related to members of the genus Rochalimaea and that the isolate is genotypically identical to the presumptive etiologic agent of bacillary angiomatosis. However, the same genotypic analyses readily differentiated the new isolate from isolates of other recognized Rochalimaea species as well as other genera of bacteria previously suggested as putative etiologic agents of bacillary angiomatosis and related syndromes. We propose that the novel species be referred to as Rochalimaea henselae sp. now.
Antimicrobial lock treatment using 40 mg/mL(-1) of tetrasodium EDTA for at least 21 hours could significantly reduce or potentially eradicate CVC-associated biofilms of clinically relevant microorganisms.
Four strains of novel, rapidly growing, acid–alcohol-fast-staining bacteria were characterized with a polyphasic approach. Isolates were received by the Centers for Disease Control and Prevention from domestic health department laboratories for reference testing as unidentifiable, clinical mycobacteria. Bacteria were rod-shaped and produced non-pigmented (white to beige), non-photochromogenic, smooth or wrinkled-rough colonies on Middlebrook 7H10 and 7H11 media at 33 °C. The smooth and wrinkled colony forms were representative of two species with 68·0 and 72·0 mol% DNA G+C content. The cell wall contained meso-diaminopimelic acid and mycolic acids. Species were characterized by cellular fatty acids of C10 : 0, C14 : 0, C16 : 1ω9t, C16 : 0, C18 : 1ω9c and 10-methyl C18 : 0 (tuberculostearic acid). HPLC analysis of mycolic acids produced a novel late-emerging, genus-specific mycolate pattern. TLC analysis demonstrated a novel α +-mycolate. Species were 98·9 % similar by comparison of 16S rRNA gene sequences; however, the DNA–DNA association was <28 %. Phylogenetic analysis of 16S rRNA gene sequences demonstrated an association with Rhodococcus equi, although a DNA–DNA relatedness value of 2 % did not support a close relationship. PCR analysis of a proposed, selected actinomycete-specific 439 bp fragment of the 65 kDa heat-shock protein was negative for three of the four isolates. The creation of Segniliparaceae fam. nov. is proposed to encompass the genus Segniliparus gen. nov., including two novel species, the type species Segniliparus rotundus sp. nov. and Segniliparus rugosus sp. nov., with the respective type strains CDC 1076T (=ATCC BAA-972T=CIP 108378T) and CDC 945T (=ATCC BAA-974T=CIP 108380T).
Central venous catheter needleless connectors (NCs) have been shown to develop microbial contamination.A protocol was developed for the collection, processing, and examination of NCs to detect and measure biofilms on these devices. Sixty-three percent of 24 NCs collected from a bone marrow transplant center contained biofilms comprised primarily of coagulase-negative staphylococci.Intravenous (i.v.) access lines (6, 7) and needleless connectors (NCs) (3, 4) have been demonstrated to be a risk factor for blood stream infection (BSI). Patients who require long-term i.v. access, such as bone marrow transplant patients, are at even greater risk for BSI. To deliver i.v. fluids (e.g., medication, blood products, or nutrients), tubing must be connected to i.v. catheters that enter the patient's bloodstream. Until recently, such connections have been made using beveled, hollow-bore needles that pierce an elastic membrane on a catheter end cap. Because of the potential for needle-stick injuries and health care worker exposure to bloodborne pathogens, many institutions have recently adopted the use of NCs. Though safer for health care workers, the potential for NCs to increase BSI risk to patients has been documented in outbreaks of nosocomial BSI (3,4). In October 1998, the Centers for Disease Control and Prevention (CDC) was asked to investigate a BSI outbreak at a bone marrow transplant center in which NCs were involved. As part of this investigation, CDC assessed the ability of NCs to harbor biofilms that could act as a reservoir for BSI pathogens. It is well established that biofilms may develop on intravascular devices, including central venous catheters (CVCs) (1,2,8). Though contamination of NCs by various organisms has been observed (3, 4), the occurrence of biofilms on these devices has, to our knowledge, not been documented. The objectives of this study were (i) to develop a standardized protocol that could be used to collect, ship, and process NCs for biofilm contamination and (ii) to determine whether biofilms could develop on these devices and what organisms were the primary colonizers. Hickman NCs were collected from patients with long-term CVCs in a single bone marrow transplant center in which an outbreak of BSIs had occurred.Collection and shipment of NCs. Female-female luer couplings (no. 06359-42; Cole Parmer, Niles, Ill.) were autoclaved and then used to connect two 5-ml syringes. One of the syringes contained 5-ml of phosphate-buffered saline (PBS; pH 7.2; Life Technologies, Grand Island, N.Y.). Syringe pairs were placed into zip-lock bags and shipped on ice packs to the bone marrow transplant center for the collection of NCs. By using an aseptic technique, the NCs were removed from the patient's CVC and placed into an unused sterile Petri dish and transported to the laboratory. After the two syringes were separated, the luer coupling remained on one syringe. The smaller end of the NC was then wiped with a sterile alcohol pledget and connected to the syringe without the luer coupling. The other end o...
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