Mycobacterium ulcerans is the causative agent of Buruli ulcer, one of the most common mycobacterial diseases of humans. Recent studies have implicated aquatic insects in the transmission of this pathogen, but the contributions of other elements of the environment remain largely unknown. We report here that crude extracts from two green algae added to the BACTEC 7H12B culture medium halved the doubling time of M. ulcerans and promoted biofilm formation. Using the 7H12B medium, modified by the addition of the algal extract, and immunomagnetic separation, we also demonstrate that M. ulcerans is associated with aquatic plants in an area of the Ivory Coast where Buruli ulcer is endemic. Genotype analysis showed that plant-associated M. ulcerans had the same profile as isolates recovered in the same region from both aquatic insects and clinical specimens. These observations implicate aquatic plants as a reservoir of M. ulcerans and add a new potential link in the chain of transmission of M. ulcerans to humans.Mycobacterium ulcerans is an emerging environmental pathogen (2). It is the etiologic agent of Buruli ulcer, a necrotic skin disease highly prevalent in many countries throughout west Africa and one of the most common mycobacterial diseases in humans after tuberculosis and leprosy (2). M. ulcerans is the only mycobacterium known to produce a toxin, a polyketide-derived macrolide called mycolactone (10, 11). The infection begins with a painless nodule or papule that spreads over the surrounding tissue. Ischemic and necrotized tissue disappears and is replaced by centralized ulceration of the limb. At present, the only effective treatment consists of excision of the lesions, and this is often followed by extensive skin grafting (6).Until very recently M. ulcerans had never been isolated in culture from the environment. Indirect evidence from epidemiological studies suggests that M. ulcerans is an environmental mycobacterium present in swampy areas. Humans are thought to be infected through minor wounds or skin abrasions via contact with mycobacterium-containing water (33). In a previous study we demonstrated that aquatic insects are a possible mode of transmission of M. ulcerans to humans. We showed that, in an experimental mouse model, aquatic insects (Naucoridae) were able to transmit an infection by biting, thereby inoculating bacilli that had accumulated in the salivary glands of the insects (16). We were then able to isolate in pure culture M. ulcerans from the salivary glands of Naucoridae captured in a region of endemicity in the Ivory Coast. However it seems unlikely that these insects are the only environmental source of M. ulcerans. In another recent study, data gathered using an M. ulcerans-specific PCR to survey environments of endemicity in southeastern Australia identified aquatic plants as a possible reservoir of this pathogen (30).In this report, we test the hypothesis that M. ulcerans is associated with aquatic plants by studying the effects of crude aquatic plant extracts on the growth of M. ulcer...

Osteoblasts are target cells for glucocorticoids and calcitriol, and their phenotype is greatly modified by these hormones. We investigated the effect of continuous or discontinuous hormonal exposure to osteoblasts derived from rat bone marrow stromal cells in long-term subcultures. Stromal cells were grown in primoculture in presence of dexamethasone (dex), but in following subcultures, dex and/or calcitriol were added just after seeding or after a 7-day hormone-free period. Cell proliferation, alkaline phosphatase (ALP) histochemical staining, and enzymatic bioactivity measurement, osteocalcin (OC), ALP and bone sialoprotein (BSP) mRNA expression were used to study the differential effect on osteoblastic phenotype of various conditions of treatment by dex and calcitriol. In primoculture, the osteoblastic differentiation was confirmed by the formation of calcified nodules and by strong expression of ALP, OC, and BSP mRNAs. In subcultures, proliferation of stromal cells was stimulated by dex and inhibited by calcitriol and by both hormones. Cell proliferation was not modified by hormonal lack during 7 days. Continuous hormonal treatment by dex strongly enhanced OC and BSP mRNAs, but apparently did not modified ALP mRNAs expression. Continuous treatment by calcitriol decreased ALP and the dex-induced BSP expression and stimulated the OC mRNAs level, strongly when associated with dex. The population of ALP+ cells and ALP bioactivity were strongly increased by dex, whereas calcitriol or both hormones decreased them. When the subcultures were undergone without hormonal treatment during 7 days, all osteogenic mRNAs strongly decreased even after hormonal recovery. Dex, calcitriol, and both hormones inhibited ALP mRNAs. OC messengers were only weakly detectable with both hormones. ALP+ cell population and ALP bioactivity were decreased after 14 days of hormonal treatment recovery. These results support that continuous presence of glucocorticoids appears as a major key for the permanent expression of the osteoblastic phenotype that is inhibited by calcitriol, in the rat bone marrow.

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