Summary: Actinobacteria constitute one of the largest phyla among Bacteria and represent gram-positive bacteria with a high G+C content in their DNA. This bacterial group includes microorganisms exhibiting a wide spectrum of morphologies, from coccoid to fragmenting hyphal forms, as well as possessing highly variable physiological and metabolic properties. Furthermore, Actinobacteria members have adopted different lifestyles, and can be pathogens (e.g., Corynebacterium, Mycobacterium, Nocardia, Tropheryma, and Propionibacterium), soil inhabitants (Streptomyces), plant commensals (Leifsonia), or gastrointestinal commensals (Bifidobacterium). The divergence of Actinobacteria from other bacteria is ancient, making it impossible to identify the phylogenetically closest bacterial group to Actinobacteria. Genome sequence analysis has revolutionized every aspect of bacterial biology by enhancing the understanding of the genetics, physiology, and evolutionary development of bacteria. Various actinobacterial genomes have been sequenced, revealing a wide genomic heterogeneity probably as a reflection of their biodiversity. This review provides an account of the recent explosion of actinobacterial genomics data and an attempt to place this in a biological and evolutionary context.

Genomics of Actinobacteria: Tracing the Evolutionary History of an Ancient Phylum

Microbial pathogens use a number of genetic strategies to invade the host and cause infection. These common themes are found throughout microbial systems. Secretion of enzymes, such as phospholipase, has been proposed as one of these themes that are used by bacteria, parasites, and pathogenic fungi. The role of extracellular phospholipase as a potential virulence factor in pathogenic fungi, including Candida albicans, Cryptococcus neoformans, and Aspergillus, has gained credence recently. In this review, data implicating phospholipase as a virulence factor in C. albicans, Candida glabrata, C. neoformans, and A. fumigatus are presented. A detailed description of the molecular and biochemical approaches used to more definitively delineate the role of phospholipase in the virulence of C. albicans is also covered. These approaches resulted in cloning of three genes encoding candidal phospholipases (caPLP1, caPLB2, and PLD). By using targeted gene disruption, C. albicans null mutants that failed to secrete phospholipase B, encoded by caPLB1, were constructed. When these isogenic strain pairs were tested in two clinically relevant murine models of candidiasis, deletion of caPLB1 was shown to lead to attenuation of candidal virulence. Importantly, immunogold electron microscopy studies showed that C. albicans secretes this enzyme during the infectious process. These data indicate that phospholipase B is essential for candidal virulence. Although the mechanism(s) through which phospholipase modulates fungal virulence is still under investigations, early data suggest that direct host cell damage and lysis are the main mechanisms contributing to fungal virulence. Since the importance of phospholipases in fungal virulence is already known, the next challenge will be to utilize these lytic enzymes as therapeutic and diagnostic targets.

Potential Role of Phospholipases in Virulence and Fungal Pathogenesis

The etiology of nosocomial infections, the frequency of contaminated hands with the different nosocomial pathogens, and the role of health care workers' hands during outbreaks suggest that a hand hygiene preparation should at least have activity against bacteria, yeasts, and coated viruses. The importance of efficacy in choosing the right hand hygiene product is reflected in the new Centers for Disease Control and Prevention guideline on hand hygiene (J. M. Boyce and D. Pittet, Morb. Mortal. Wkly. Rep. 51:1-45, 2002). The best antimicrobial efficacy can be achieved with ethanol (60 to 85%), isopropanol (60 to 80%), and n-propanol (60 to 80%). The activity is broad and immediate. Ethanol at high concentrations (e.g., 95%) is the most effective treatment against naked viruses, whereas n-propanol seems to be more effective against the resident bacterial flora. The combination of alcohols may have a synergistic effect. The antimicrobial efficacy of chlorhexidine (2 to 4%) and triclosan (1 to 2%) is both lower and slower. Additionally, both agents have a risk of bacterial resistance, which is higher for chlorhexidine than triclosan. Their activity is often supported by the mechanical removal of pathogens during hand washing. Taking the antimicrobial efficacy and the mechanical removal together, they are still less effective than the alcohols. Plain soap and water has the lowest efficacy of all. In the new Centers for Disease Control and Prevention guideline, promotion of alcohol-based hand rubs containing various emollients instead of irritating soaps and detergents is one strategy to reduce skin damage, dryness, and irritation. Irritant contact dermatitis is highest with preparations containing 4% chlorhexidine gluconate, less frequent with nonantimicrobial soaps and preparations containing lower concentrations of chlorhexidine gluconate, and lowest with well-formulated alcohol-based hand rubs containing emollients and other skin conditioners. Too few published data from comparative trials are available to reliably rank triclosan. Personnel should be reminded that it is neither necessary nor recommended to routinely wash hands after each application of an alcohol-based hand rub. Long-lasting improvement of compliance with hand hygiene protocols can be successful if an effective and accessible alcohol-based hand rub with a proven dermal tolerance and an excellent user acceptability is supplied, accompanied by education of health care workers and promotion of the use of the product.

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Epidemiologic Background of Hand Hygiene and Evaluation of the Most Important Agents for Scrubs and Rubs

Adherence and receptor relationships of Candida albicans.

Candida albicans is part of the normal human flora, and it grows on mucosal surfaces in healthy individuals. In susceptible hosts, this organism can cause both mucosal and hematogenously disseminated disease. For C. albicans to persist in the host and induce disease, it must be able to adhere to biotic and abiotic surfaces, invade host cells, and obtain iron. The C. albicans hypha-specific surface protein Als3 is a member of the agglutinin-like sequence (Als) family of proteins and is important in all of these processes. Functioning as an adhesin, Als3 mediates attachment to epithelial cells, endothelial cells, and extracellular matrix proteins. It also plays an important role in biofilm formation on prosthetic surfaces, both alone and in mixed infection with Streptococcus gordonii. Als3 is one of two known C. albicans invasins. It binds to host cell receptors such as E-cadherin and N-cadherin and thereby induces host cells to endocytose the organism. Als3 also binds to host cell ferritin and enables C. albicans to utilize this protein as a source of iron. Because of its multiple functions and its high expression level in vivo, Als3 is a promising target for vaccines that induce protective cell-mediated and antibody responses. This review will summarize the multiple functions of this interesting and multifunctional protein.

Candida albicans Als3, a Multifunctional Adhesin and Invasin

In oral candidiasis, many keratinized epithelial cells and cells of Candida albicans are shed. Scales from patients with oral candidiasis were used for electron microscopic study of the epithelial-fungal relationship. Scales, scraped from the tongue and oral mucosa, were fixed for fungi. Electron microscopic observations showed cells of C. albicans outside, penetrating, or within the epithelial cells. Extracellular fungi possessed a floccular material adherent to the outer surface of the cell wall. Intracellular fungi lacked the floccular material which appeared to detach as fungi invaded the epithelial cells. Large vacuoles, which sometimes contained myelin figures, occupied the cytoplasm of fungal cells. Epithelial cells frequently contained several fungi. Discontinuous plasma membranes marked sites of fungal entry. Cytoplasmic areas devoid of fungi showed many tonofibrils, but the cytoplasm adjacent to fungi often lacked tonofibrils. Micrographs suggested that fungal cells lysed the tonofibrils. Bacteria were abundant in the scrapings, but always occupied an extracellular position.

Ultrastructural features of host-parasite relationship in oral candidiasis.

the past 30 years knowledge of cutanef)ua bacteriology lias gradually accumulated (Marples, M. J., 1965). Yet, one main aspect of cutaneous bacterial flora has not been completely elucidated, i.e., the site where bacteria are normally found in the epidermis. An investigation was undertaken to visualize by means of correlative light and electron microscopy the location of normal skin flora. Results of this study are presented.

Location of bacterial skin flora.

Fine Structure of Corynebacterium Acnes

Simultaneous occurrence of subacute sclerosing panencephalitis (SSPE) was documented in two brothers by clinical, histopathologic, electronmicroscopic, immunologic, and virologic studies. Other intrafamilial cases were reviewed, and the implications of all cases of this type were discussed in light of modern concepts of the pathogenesis of SSPE. The findings in studies of such cases cast doubt on the theory of simple random reactivation of measles as a cause of SSPE and suggest that environmental factors may play a role in initiation of the disease.

https://www.jstor.org/stable/pdfplus/30111554.pdf

Walter H. Wilborn

Papers

Ultrastructural features of host-parasite relationship in oral candidiasis.

Location of bacterial skin flora.

Fine Structure of Corynebacterium Acnes

The Simultaneous Occurrence of Subacute Sclerosing Panencephalitis in Two Brothers. I. Clinical, Virologic, and Histopathologic Studies