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Phylogenetic analysis identifies the invertebrate pathogen Helicosporidium sp. as a green alga (Chlorophyta).

Historically, the invertebrate pathogens of the genus Helicosporidium were considered to be either protozoa or fungi, but the taxonomic position of this group has not been considered since 1931. Recently, a Helicosporidium sp., isolated from the blackfly Simulium jonesi Stone & Snoddy (Diptera: Simuliidae), has been amplified in the heterologous host Helicoverpa zea. Genomic DNA has been extracted from gradient-purified cysts. The 185, 28S and 5.8S regions of the Helicosporidium rDNA, as well as partial sequences of the actin and beta-tubulin genes, were amplified by PCR and sequenced. Comparative analysis of these nucleotide sequences was performed using neighbour-joining and maximum-parsimony methods. All inferred phylogenetic trees placed Helicosporidium sp. among the green algae (Chlorophyta), and this association was supported by bootstrap and parsimony jackknife values. Phylogenetic analysis focused on the green algae depicted Helicosporidium sp. as a close relative of Prototheca wickerhamii and Prototheca zopfii (Chlorophyta, Trebouxiophyceae), two achlorophylous, pathogenic green algae. On the basis of this phylogenetic analysis, Helicosporidium sp. is clearly neither a protist nor a fungus, but appears to be the first described algal invertebrate pathogen. These conclusions lead us to propose the transfer of the genus Helicosporidium to Chlorophyta, Trebouxiophyceae.

Actins↗

Two cases of protothecosis in Nagoya, Japan.

Two cases of protothecosis caused by Prototheca wickerhamii have been reported from Nagoya in a 12 year period in both cases the infection presented on the cheeks of otherwise healthy women. Biopsies showed numerous PAS positive staining organisms with the distinctive mulberry like endosporulation in the dermis. Prototheca wickerhamii was identified on sugar assimilation tests of colonies isolated from tissue on Sabouraud agar. Case 1 responded to 11 months of oral ketoconazole therapy. Case 2 might not respond to itraconazole. The source of the infections has not been identified.

Adult↗

[Generalized protothecosis in a fruit bat (Pteropus lylei)].

Prototheca organisms, histologically resembling Prototheca wickerhamii, were found in a fruit bat as a cause of a massive granulomatous lymphadenitis and splenitis and a widespread granulomatous meningoencephalitis. Focal lesions were also present in the myocardium, in skeletal muscles, and in the kidneys.

Animals↗

Susceptibilities against bovine lactoferrin with microorganisms isolated from mastitic milk.

Antibacterial effects of bovine lactoferrin were studied in vitro against microorganisms isolated from mastitic milk in Tokachi area, Hokkaido, Japan. Microorganisms isolated were Escherichia coli (11 isolates), Klebsiella pneumoniae (5 isolates), enterococci (8 isolates), Staphylococcus aureus (10 isolates), coagulase negative staphylococci (CNS, 13 isolates), streptococci (11 isolates), Prototheca zopfii (7 isolates) and yeast-like fungi (9 isolates). Lactoferrin has been known as a multifunctional protein and its antimicrobial effect is one of the most essential function of it. In order to compare their susceptibilities against lactoferrin, the minimal inhibitory concentration values were estimated by a microplate assay method using 96-well microplate, which involved measuring the optical density of the cultures. Prototheca zopfii was highly sensitive to bovine lactoferrin and complete inhibition of this microorganism was observed even at the low concentration of 7 mug/ml. On the other hand, E. coli and enterococci showed resistance against lactoferrin action and staphylococci showed strain-dependent resistance.

Animals↗

Papular protothecosis of the chest. Immunologic evaluation and treatment with a combination of oral tetracycline and topical amphotericin B.

A 65-year-old woman presented with cutaneous, pruritic papules of the upper part of her chest. Histologic findings and culture of a biopsy specimen confirmed that these papules were caused by an infection with Prototheca wickerhamii, an achloric alga that rarely infects humans. Natural killer cell activity in this patient was significantly depressed before initiation of therapy. Reevaluation following successful treatment with oral tetracycline hydrochloride and topical amphotericin B cream revealed an improvement in natural killer cell activity. This patient is presented to demonstrate a rare clinical presentation of protothecosis; to document that cutaneous protothecosis can be successfully treated without the risk of toxic side effects associated with systemic antifungal agents; and to illustrate that although most patients with cutaneous protothecosis are immunosuppressed, infection with this alga appears to be at least partially responsible for the immunosuppression.

Administration, Topical↗

In vitro antimycotic activity of some plant extracts towards yeast and yeast-like strains.

As part of screening aimed at the selection of novel antimycotic compounds of vegetable origin, leaf extracts of Camellia sinensis L., Cupressus sempervirens L. and Pistacia lentiscus L. and the seed extract of Glycine soja Sieb. et Zucc. were tested against yeast and yeast-like species implicated in human mycoses. Of the extracts only those of C. sinensis (obtained from a commercial preparation of green tea) exhibited broad activity towards Candida glabrata, Clavispora lusitatiae, Cryptococcus laurentii, Filobasidiella neoformans, Issatchenkia orientalis, Saccharomyces cerevisiae and Prototheca wickerhamii strains. MICs ranging from 300 to 4800 microg extract/mL (corresponding to 130-2010 microg/mL total polyphenols) were observed. Concentrations of the C. sinensis extract over 25 000 microg/mL caused a rapid decrease of viable cells of Fil. neoformans and its activity was dose-dependent. Tests carried out using the pure polyphenols present in C. sinensis extract composition, showed that only epicatechin-3-O-gallate (ECG) and epigallocatechin-3-O-gallate (EGCG) possess antimycotic activity.

Antifungal Agents↗

Protothecosis: a report of two cases in Japan and a review of the literature.

Protothecosis is an emerging opportunistic infection caused by species belonging to the genus Prototheca. Two Japanese cases of protothecosis are documented with a critical review of the literature. A current perspective concerning the microbiology and disease entity of protothecosis is described in detail.

Adolescent↗

A group-I intron in the mitochondrial large-subunit ribosomal RNA-encoding gene of Dictyostelium discoideum: same site localization in alga and in vitro self-splicing.

A 547-bp group-I intron belonging to subgroup IA1 was found near the 3' end of the large subunit ribosomal RNA-encoding gene (LSUrRNA) in the mitochondrial (mt) DNA of the cellular slime mold Dictyostelium discoideum. This intron was inserted in a highly conserved stretch within the sequence that encodes the peptidyl transferase center domain V in the corresponding region of the Escherichia coli LSUrRNA. Interestingly, the insertion site of the intron is the same as that of the So.LSU.2 intron of the green alga, Scenedesmus obliquus, mt DNA and the Pw.LSU.2 intron of the colorless alga, Prototheca wickerhamii, mt DNA. The intron could self-splice in vitro at a concentration higher than 20 mM MgCl2. Polymerase chain reaction analysis showed the possible existence of an intron similar to that of D. discoideum LSUrRNA in another cellular slime mold, Polysphondylium pallidum (CK-8), but not in D. mucoroides (Dm7 and Dm11).

Animals↗

Canine cutaneous protothecosis: an immunohistochemical analysis of the inflammatory cellular infiltrate.

In this immunohistochemical study, the distribution of the cellular inflammatory infiltrate associated with cutaneous canine protothecosis (Prototheca wickerhamii) was evaluated by consecutive biopsies taken before, during and after treatment. Antibodies specific to canine immunoglobulins (IgG, IgM, IgA), human CD3 antigen (pan T-lymphocyte marker) and human myeloid/ histiocyte antigen (macrophage/neutrophil marker) were used. Before treatment, cellular infiltrate was very scanty in the inflamed areas, but it increased during the treatment, whereas the number of protothecal organisms decreased. Statistical analysis revealed an inverse relation between the number of protothecal organisms and the number of infiltrating macrophage/neutrophils (P < 0.004), T lymphocytes (P < 0.001), and cells containing immunoglobulin G (P < 0.001), M (P < 0.001) and A (P < 0.001) at different stages of the disease. These findings suggest either that protothecal organisms inhibit the migration or proliferation of cellular inflammatory infiltrate or that only dead protothecal organisms induce an effective local immune response.

Animals↗

Protothecosis.

Intravenous amphotericin B remains the most effective drug for eradicating Prototheca infections. It should be used as a first-line agent in cases of disseminated disease and in patients with severe underlying illness or with immunosuppression or immunocompromise. Azole antifungals and surgery should be reserved for patients with more localized disease. Itraconazole appears to be the most effective agent of this drug class, and it should be administered at 200 mg/day for 2 months. Surgical excision should be considered as a first-line therapy in patients who present with olecranon bursal infections.

Amphotericin B↗

Systemic chlorellosis, an emerging infection in humans caused by algae.

One hundred and eight cases of human protothecosis occurring over 25 years are reviewed. The most common species isolated was Prototheca wickerhamii, The algae have been isolated from a large number of sites but seem to have low virulence. In the majority of systemic infections there have been few symptoms. Cutaneous manifestations may vary from patient to patient but infections of immunosuppressed patients tend to be more florid. Most cases have been successfully treated by excising the lesion or with antifungals. Most strains are sensitive to 5-fluorocytosine, amphotericin B and the azoles but resistance does occur.

Chlorella↗

4,4,14 alpha-trimethyl 9 beta,19-cyclo-5 alpha-26-homocholesta-24,26-dien-3 beta-ol: a potent mechanism-based inactivator of delta 24(25)- to delta 25(27)-sterol methyl transferase.

The title compound (4A) was synthesized and tested as a mechanism-based inactivator of the sterol methyl transferase (SMT) enzyme from Prototheca wickerhamii. Using cycloartenol as substrate, 4A was found to exhibit time-dependent inactivation kinetics, generating a Ki value of 30 microM and Kinact value of 0.30 min-1.

Cholesterol↗

Cutaneous protothecosis: report of five cases.

Prototheca, a genus of achlorophyllic algae, is a rare cause of opportunistic infection in humans. About 80 human cases, mostly cutaneous infection caused by P. wickerhamii, have been reported world-wide. We describe the clinicopathological findings and treatments of five cases diagnosed in our department during the period 1991-97. The patients, including four elderly farmers, presented with pyoderma-like lesions or infiltrating papules and plaques on the extensor side of the extremities or face. All patients were immunocompromised, mostly due to systemic or topical steroids. The diagnosis was made in each case by finding typical endospores with morula-like structures in skin biopsy specimens. P. wickerhamii was isolated in four cases in which pretreatment culture of skin tissue was done. Except for one patient who died of asthma, the infection was cured after 2-7 weeks of amphotericin B, ketoconazole, itraconazole or fluconazole. Our cases illustrate that cutaneous protothecosis commonly manifested non-tender, pyoderma-like or infiltrating lesions and should be considered in the differential diagnosis of deep fungal or mycobacterial infection. This rare infection seemed more frequent in our region, possibly due in part to common steroid abuse among old people in Taiwan and there was a large population of elderly farmers in our area.

Adult↗

Aetiology and histopathology of bovine mastitis of espontaneous occurrence.

Infectious bovine mastitis is responsible for serious economic losses in dairy cattle breeding. Information about the damages caused by micro-organisms is of great interest and importance. The purpose of the present study was to report on the microbiological and histopathological aspects of the mammary parenchymas of slaughtered dairy cows. A total of 184 mammary glands were examined and samples of mammary parenchyma were collected for microbiological and histopathological examinations. Micro-organisms were isolated from 69.6% samples; 23 (12.5%) of the 184 samples did not show histological changes; inflammatory response was observed in 56 (30.4%) samples; inflammatory response and repair were present in 82 (44.6%) samples; repair process was verified in 23 (12.5%) of the mammary glands. Coagulase-negative Staphylococci were observed in 53.8% of the samples followed by coagulase-positive Staphylococci (7.6%), Prototheca sp. (2.2%) and Streptococcus dysgalactiae (1.6%). Coagulase-negative Staphylococci and coagulase-positive Staphylococci were associated mainly to chronic inflammatory response and chronic inflammatory response and repair. Samples from which no micro-organisms were isolated (n = 56) had no histological changes in 82.6% of the cases. These results were higher (P < 0.05) when compared to the samples with micro-organisms and without histological changes (17.4%).

Animals↗

Protothecosis.

Protothecosis is an infection caused by achlorophyllic algae of the genus Prototheca which rarely affects humans. Some 100 cases have been described in the medical literature, the majority caused by the species P. wickerhamii. The skin is the organ most frequently involved. Diagnosis is performed by isolation of the microorganism in culture or by histopathology. The ideal treatment has not been defined, with amphotericin B and the azoles having been employed. Surgical excision is recommended for small, localized lesions. We describe a case of cutaneous protothecosis on the right fourth finger of a female patient 59 years old with no underlying disease. Administration of itraconazole 400 mg/day for 6 weeks failed to produce an adequate clinical response. Treatment was then changed to fluconazole 200 mg/day, with regression of the lesion.

Amphotericin B↗

Protothecosis complicating prolonged endotracheal intubation: case report and literature review.

Prototheca species are ubiquitous, aerobic, unicellular algae closely related to the green algae Chlorella. Their involvement in human disease--in both immunocompetent and immunocompromised patients--has been reported with increasing frequency. The wide array of presentations has included cutaneous, subcutaneous, mucosal, bursal, catheter-related, and (in rare instances) systemic disease. We report a case of protothecosis complicating prolonged endotracheal intubation presenting as a nasopharyngeal ulceration with a soft-tissue mass, and we review the presentation, treatment, and outcome of the 59 previously reported cases of protothecosis. Optimal therapy for protothecosis includes excision (where possible) followed by systemic administration of amphotericin B; the sole exception is in the case of olecranon bursitis, where excision alone appears curative. The role of the newer imidazoles is yet to be determined.

Adult↗

Visceral protothecosis mimicking sclerosing cholangitis in an immunocompetent host: successful antifungal therapy.

A healthy 39-year-old man who had clinical findings consistent with sclerosing cholangitis was found to have systemic protothecosis at surgery. Severe granulomatous inflammation and palpable nodules were found in the gallbladder, on the surface of the liver, and in the duodenum. Prototheca wickerhamii was detected in biopsied specimens and stool; the titer of indirect fluorescent antibody to this organism was 1:2,000. The patient recovered after a short course of treatment with amphotericin B and 3 months of oral therapy with ketoconazole. He had no other concurrent illness and had no abnormality in his immune system. This is the second reported human case of systemic protothecosis. An elevated IgG level, an elevated erythrocyte sedimentation rate, eosinophilia, and abnormal levels of enzymes in the liver were found in both cases. Protothecosis should be considered in the differential diagnosis of hepatic and biliary inflammatory diseases of uncertain etiology.

Adult↗

Protothecal olecranon bursitis: treatment with intrabursal amphotericin B.

A patient is described from whom repeated olecranon bursa fluid cultures yielded Prototheca wickerhamii. Following intrabursal instillation of amphotericin B, the cultures became negative. There was no progression of infection or adverse consequences of this treatment. Review of the literature regarding this unusual infection reveals seven reported cases of protothecal olecranon bursitis. Five cases from which sufficient information was provided were diagnosed by histologic examination of excised bursa that showed the characteristic sporangia.

Amphotericin B↗