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Ultrastructure of Prototheca zopfii in bovine granulomatous mastitis.

Mammary glands from cows with protothecal mastitis were examined by light and electron microscopy at 6, 13, 20, and greater than 180 days after infection. With increasing time, there were increases in severity of granulomatous inflammation, number of endospores and sporangia, and ratio of degenerate to intact algae. Algae were found in macrophages but were not seen in neutrophils, epithelial cells, or myoepithelial cells. Macrophages containing algae were markedly enlarged, chiefly from reduplication of the Golgi complex and its associated vesicles. Intracellular algae were degenerate and consisted of intact cell wall profiles which contained membrane fragments but lacked nuclei and cytoplasmic organelles. Degenerate algae in vitro had thin cell walls and did not undergo internal lysis. Cell wall material of intracellular algae stained as an acidic, nonsulfated, carboxylated glycoprotein. These findings suggest that intracellular Prototheca zopfii degenerate by progressive lysis of internal organelles with persistence of cell wall glycans and that development of aberrant cell wall forms occurs as a defective response by host macrophages.

Animals↗

Serologic survey of rotavirus, Norwalk agent and Prototheca wickerhamii in wastewater workers.

Analysis of paired sera from 48 wastewater workers and controls who reported gastrointestinal illness did not reveal any excess of seroconversions to Norwalk agent or to rotavirus. Inexperienced wastewater-exposed workers had higher levels of antibody to Norwalk agent than did experienced and control workers and those with high and medium aerosol exposure had higher titres than those in the low aerosol category. Analysis for Prototheca antibody titres was essentially negative.

Adult↗

Chronic Prototheca meningitis.

A case of chronic meningitis caused by the achloric alga Prototheca wickerhamii is described, which has persisted for more than 6 years despite treatment with various antifungal agents. For the last year no treatment has been given, but the patient has no complaints.

Adult↗

[Individual animal risk factors for Prototheca mastitis in cattle].

Risk factors for mastitis caused by Prototheca zopfii were determined in a retrospective case control study. 248 cows culled for protothecal mastitis in a large dairy herd were included in the study. The diseased cows were compared to their non affected herdmates in respect to age, stage of lactation, production and milk composition. The effect of previous cases of mastitis during the same lactation or in previous lactations and the effect of antibiotic pretreatment were determined in a split udder analysis comparing left quarters to right quarters. Age and mastitis history especially antimicrobial pretreatment were important risks factors for developing protothecal mastitis. Risk of protothecal mastitis increased with parity. Quarters with a history of clinical mastitis during the same or during previous lactations were at higher risk than other quarters (OR 1.98, p < 0.01 and 1.44, p = 0.06, respectively). Pretreatment with antibiotics increased the risk of protothecal mastitis (OR 3.19, p < 0.001). Milk yield was lower in cows that developed protothecal mastitis. Somatic cell counts were higher than in control cows. Incidence of protothecal mastitis peaked in the second month of lactation.

Age Factors↗

Antifungal drug combinations for Cryptococcus neoformans and Prototheca spp.

Seventy-one isolates of Cryptococcus neoformans and 5 isolates of Prototheca spp. were tested for in vitro susceptibility against amphotericin B alone and against the combination of amphotericin B with each clinically relevant concentration of flucytosine (5-FC) and rifampin by broth dilution methods. The combinations of amphotericin B and rifampin produced greater effect on reduction of the minimal inhibition concentration (MIC) of amphotericin B than did either drug used individually. Flucytosine combined with amphotericin B produced little or no reduction of the MIC compared with amphotericin B alone.

Amphotericin B↗

[Antifungal effect of three natural products on the genetic substance of Saccharomyces cerevisiae GL7 and Prototheca wickerhamii].

AIM: To investigate the antifungal effect of three natural products on the genetic substance of Saccharomyces cerevisiae GL7 and Prototheca wickerhamii. METHODS: The normal and treated cells were observed by confocal laser scanning microscope (CLSM) and image analysis to quantitatively described the cell morphology, area, DNA and RNA content. RESULTS: The morphology, area, DNA and RNA contents were changed greatly in the treated cells. CONCLUSION: Solasodine, 4'-hydroxy-3, 5-dimethoxystilbene and dictamnine directly or indirectly interfered the synthesis and function of genetic substance in S. cerevisiae and P. wickerhamii.

Alkaloids↗

Bovine mastitis due to Prototheca zopfi.

A report is given of a case of bovine mastitis caused by the alga Prototheca zopfi which was erroneously identified as the yeast Cryptococcus neoformans. The resistance of protothecal mastitis to treatment with antibiotics commonly used in veterinary practice is briefly discussed.

Animals↗

Natural and experimental bovine intramammary infection with Prototheca zopfii.

A naturally occurring case of chronic mastitis caused by Prototheca zopfii was detected in a dairy cow. The cow was observed for 12 weeks, and milk samples were cultured each week. Milk production was decreased, and high numbers (greater than 10(3)/ml) of the organism were detected in the milk. Six glands of 2 cows were inoculated with small numbers of P zopfii. All 3 cows were euthanatized, and at postmortem, a progressive pyogranulomatous inflammation was observed in all infected mammary glands. The organism was recovered from tissue and secretions from all inoculated glands and from lymph nodes draining these glands but not from other tissues.

Animals↗

Antimicrobial susceptibility of Prototheca zopfii isolated from bovine intramammary infections.

In vitro antimicrobial susceptibility tests were carried out on 48 strains of Prototheca zopfii, an achlorophyllous algae causing refractory mastitis in dairy cows; 27 antimicrobials were evaluated. All strains were susceptible to both myxin and nystatin. In addition, 22 strains were susceptible to amphotericin B, 21 to polymyxin B, and 18 to gentamicin. Only 1 strain was susceptible to kanamycin. All strains were resistant to ampicillin, bacitracin, carbenicillin, cephalothin, chloramphenicol, clotrimazole, cloxacillin, erythromycin, flucytosine, ketoconazole, lincomycin, miconazole, neomycin, nitrofurazone, novobiocin, oleandomycin, penicillin, rifampin, streptomycin, tetracycline, and vancomycin.

Animals↗

Bovine mammary protothecosis due to Prototheca zopfii.

Mastitis due to Prototheca zopfii was diagnosed in three of 28 cows in a dairy herd. As two cows continued to shed algae after 45 days they were slaughtered and organs were examined by cultivation, histology, immunohistochemistry and electron microscopy. Algae were restricted to the mammary glands and regional lymph nodes in which a granulomatous inflammation was seen. Algae were predominantly seen in macrophages but neutrophils also contained organisms. In macrophages both sporangiospores and sporangia were found, suggesting that intracellular proliferation may be responsible for the failure to overcome the infection. Serum samples from all cows were assayed for antibodies against P. zopfii in an enzyme-linked immunosorbent assay (ELISA). Although the highest titre was found in an infected cow the difference between the mean values of the titre in infected and non-infected cows was not significant.

Animals↗

[Endocarditis due to an alga of the genus Prototheca sp. A saprophyte of water and of tree sap?].

We report a case of a preterm neonate with very low birth weight (775 grs), He stayed 77 days in a neonatal care unit due to respiratory distress syndrome, with respiratory support for 27 days, and complications in gastrointestinal and nervous systems. In a structural normal heart, he developed infective endocarditis that was treated with good bacteriologic results. An echocardiogram showed the persistence of a mass in the right atrium, the mass was a thrombus and it was resected. Histopathological analysis and cultures of the thrombus reported Prototheca sp. that is an algae. The patient received medical treatment with amphothericin B with satisfactory clinical evolution. He is doing well 6 months later. This is the first report of literature of an algae causing endocarditis.

Amphotericin B↗

Identification and characterization of mitochondrial translation products in various yeasts and in Prototheca zopfii.

Mitochondrial genomes of different eucaryotes are not all alike. We have examined mitochondrial translation products in a number of yeasts (Candida krusei, Hansenula saturnus, Rhodotorula glutinis, Rhodotorula rubra, Torulopsis glabrata and Saccharomyces cerevisiae) and in Prototheca zopfii, a colorless alga, in order to determine whether certain proteins are invariably synthesized within mitochondria, how different these proteins are, and what additional proteins, if any, might be synthesized by diverse mitochondria. Using a variety of techniques and criteria, including immunological analysis and peptide mapping, we show that all the yeasts studied, and probably P. zopfii as well, make versions of the 3 large subunits of cytochrome c oxidase. Not all of these oxidase subunits are equally closely related to their counterparts in S. cerevisiae, however. Mitochondria of some of the yeasts studied do not make, or make only small amounts of, a counterpart to Varl, a major mitochondrially made protein in S. cerevisiae. Mitochondria of P. zopfii possibly do not make an apocytochrome b. T. glabrata, H. saturnus and the two Rhodotorula species each make one or more proteins whose relationship, if any, to mitochondrial translation products of S. cerevisiae is not apparent. These results provide new information about mitochondrial diversity. Whereas mitochondria of all the organisms that we have studied devote the major part of their synthetic effort to making the three large subunits of cytochrome c oxidase, and probably make certain other proteins in common, they do not all synthesize a completely identical set of proteins.

DNA, Mitochondrial↗

The ultrastructure of Prototheca wickerhamii.

Two clinical isolates from Prototheca wickerhamii were freeze-dried, fixed and studied by electron microscopy and were also examined growing in culture medium by phase contrast light microscopy. Resting spores placed on fresh medium developed cytoplasmic extensions which sequestrated before proliferation occurred. In the presence of adequate nutrients vegetative spores grew and subdivided to form up to 12 endospores within large sporangia which ruptured to release free spores every 5--6 hours. These proliferating or vegetative spores contained much more endoplasmic reticulum and mitochondria than the resting spores which contained more lipid, and often starch granules as well, but relatively few membranous organelles.

Cell Membrane↗

Peroxidase activity in mitochondria of Prototheca moriformis.

Peroxidase activity was investigated by the use of diaminobenzidine method in fixed cells of Prototheca moriformis. A strong peroxidase activity was observed in the mitochondria. DAB staining was unaffected by KCN, aminotriazole and antimycin A, but it was completely inhibitied by methanol-nitroferricyanide.

Animals↗

Innate immune reactions stimulated by a lipopolysaccharide-like component of the alga Prototheca (strain 289).

We report on the influence of an LPS-like molecule (aLPS) from the pathogenic alga, Prototheca (strain 289) on insect and murine innate immune reactions. Insect innate reactions to infection include nodule formation, a process of entrapping bacterial cells in aggregates of hemocytes. We recorded eicosanoid-dependent, dose-related nodulation reactions to aLPS in hornworms (Manduca sexta). The insect reaction was attenuated by pre-incubating the aLPS with polymyxin-B. Conversely, the murine macrophages reacted to challenge with Escherichia coli LPS by secreting cytokines, but did not react to aLPS. We infer that, while highly conserved with respect to intracellular mechanisms of interaction, insect and mammalian immune surveillance systems differ in recognition of LPS molecular types.

Animals↗

Development of a novel SNV-based genotyping method for Prototheca bovis.

We developed a novel single nucleotide variant (SNV)-based genotyping method for Prototheca bovis based on mitochondrial and plastid genome polymorphisms. Sequencing of five PCR amplicons targeting six informative SNVs discriminated field isolates with a Simpson's D value of 0.927, providing a simple and robust epidemiological typing method.

Journal Article↗

The purification and properties of malonic semialdehyde oxidative decarboxylase from Prototheca zopfii.

1. An enzyme, which in the presence of NAD(+) and CoA oxidizes malonic semialdehyde to acetyl-CoA, has been purified from an extract of the colourless alga Prototheca zopfii. 2. The purified enzyme has optimum pH7.5, is specific for NAD(+) and requires a thiol compound for maximum activity. 3. The enzyme is inhibited by arsenite, N-ethylmaleimide and urea. 4. The results are discussed in relation to those obtained by other workers with a similar bacterial enzyme, and a possible reaction sequence is proposed.

Arsenicals↗