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Epithelioid cell granulomas experimentally induced by prototheca in the skin of mice: a light microscopic study.

Prototheca wickerhamii, an achlorophyllous algae, was previously found to induce massive epithelioid cell granulomas in the skin of mice. By means of light microscopy, examination was made of the histological reactions involved in epithelioid cell granulomas induced by intradermal and/or subcutaneous inoculation of Prototheca wickerhamii in BALB/c and ICR mice. Six BALB/c mice showed granuloma nodules while only three of six ICR mice did so. Based on the results of the present and previous studies, BALB/c mice may be considered a strain particularly vulnerable to contracting epithelioid cell granuloma and ICR mice, a resistant strain. In very early lesions at one week following initial prototheca inoculation, cellular infiltration with varying numbers of polymorphonuclear leukocytes, lymphocytes and some macrophages was observed throughout the dermis and subcutaneous fat tissue. In early lesions at one to two months after inoculation, focal granulomas composed of histiocytic cells and/or macrophages were observed. Mast cells were occasionally present among the histiocytic cell infiltrates. In the granulomatous lesions at two to three months, scattered eosinophils and some lymphocytes were seen. Central necrosis, with numerous neutrophils and many endospores surrounded by the granuloma, was often observed. In late stage lesions at six months, massive lymphocyte and plasma cell infiltration surrounding and/or intervening between vacuolated epithelioid cell clusters was evident. Histological reactions in epithelioid cell granuloma and the ultimate course of this disease can be staged from the histological point of view as follows: 1) diffuse inflammation, 2) cell proliferation leading to epithelioid cell formation, 3) hypertrophy of epithelioid cells with consequent formation of cell aggregates and/or organized granuloma and 4) degeneration of granuloma.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

[Udder compatibility of tetramisole and levamisole hydrochloride and suggestion for the prescription of their intramammary use in cattle against Prototheca zopfii].

Preparations with 4 percent Tetramisole or Levamisole hydrochloride were compatible in the mammary gland after application up to 40 ml/quarter (4 mg/kg body mass) for up to 6 milking times. 100 ml of the preparations were also clinically compatible in a single quarter (10 mg/kg body mass) on several days. After intramammary application of 30 ml/quarter (3 mg/kg body mass) Prototheca suppressing values in milk serum were evident for at least 6 hours. One application of 200 ml in one quarter has caused Prototheca suppressing values for 36 hours in milk serum up to dilution 1:4. Nilverm given to cattle in high dosage orally did not lead to Prototheca suppressing values in milk sera.

Animals

[Pilot study of the occurrence of Prototheca in fecal samples of horses].

Fecal samples of 146 horses were culturally investigated for occurrence of Prototheca spp. using a selective medium, developed by PORE (1973). Prototheca zopfii could be isolated in monoculture from 9 fecal samples (6.2%). The results of this pilot study show that horses too can harbour and shed Prototheca spp. in variable frequency.

Animals

[The occurrence of Prototheca in fecal samples of cattle].

Prototheca spp. were detected in 146 (48.7%) of culturally examined fecal samples of cattle, using the selective medium, developed by Pore (1973). 114 fecal samples (78.1%) contained a monoculture of Prototheca (P.) zopfii, while 10 fecal samples (6.8%) contained a monoculture of P. moriformis. In further 22 fecal samples (15.1%) a mixed culture of P. zopfii and P. moriformis was detected. The results of this study permit to conclude that cattle can harbour and shed Prototheca spp. in variable frequency.

Animals

The structural analysis of the mitochondrial SSUrRNA implies a close phylogenetic relationship between mitochondria from plants and from the heterotrophic alga Prototheca wickerhamii.

The gene for the mitochondrial small subunit rRNA (SSUrRNA) from the heterotrophic alga Prototheca wickerhamii has been isolated from a gene library of extranuclear DNA. Sequence and structural analyses allow the determination of a secondary structure model for this rRNA. In addition, several sequence motifs are present which are typically found in SSUrRNAs of various mitochondrial origins. Unexpectedly, the Prototheca RNA sequence has more features in common with mitochondrial SSUrRNAs from plants than with that from the green alga Chlamydomonas reinhardtii. The phylogenetic relationship between mitochondria from plants and algae is discussed.

Base Sequence

Prototheca associated with banana.

Prototheca stagnora was found to be a habitant of older harvested banana (Musa sapientum) and plantain (M. paradisiaca) stumps while P. wickerhamii colonized fresh Musa sp. stumps and flower bract water of Heliconia sp. While Prototheca sp. were known to habituate woody plants, this is the first evidence that herbaceous plants also serve as habitats.

Fruit

The effect of amphotericin B on the ultrastructure of Prototheca species.

Prototheca zopfii and prototheca wickerhamii stains were exposed to subinhibitory concentrations of the antimycotic amphotericin B, and the effect of the treatment on their ultrastructure was assessed. The results revealed ultrastructural changes in the treated cells, expressed by swelling of mitochondria, degradation of cell organelles, accumulation of microbody like structures, lipid droplets and starch granules in the cytoplasm, and changes in the inner layer of the cell wall.

Amphotericin B

Prototheca zopfii: natural, transient, occurrence in pigs and rats.

Domestic swine faeces and fresh faeces from trapped barnyard rats were heavily contaminated with Prototheca zopfii, a cause of dairy cow mastitis. When the pigs and rats were maintained on Prototheca-free diets, the transient intestinal population of P. zopfii decreased precipitously and disappeared. When combined with the information that other farm animals excrete P. zopfii, it was concluded that contaminated animal feed may be the source of large numbers of P. zopfii in the farm environment. We found P. zopfii in wet spoiled feed. Rats are logical vectors for contamination of feed.

Animal Feed

Cutaneous manifestations of Prototheca infections.

The genus Prototheca comprises several species, the most prevalent of which is Prototheca wickerhamii. These achlorophyllic algae are found in the slime flux of trees and in freshwater environs. Cutaneous infection with these microbes was first reported 30 years ago. Subsequent case reports have described other manifestations including systemic disease. Asymptomatic and stationary cutaneous plaques may be seen in otherwise healthy patients. Susceptibility to infection is ill defined but may involve an inability of neutrophils to effectively eradicate phagocytosed organisms. Characteristic morula are seen histologically. Antifungal medications are the most effective therapy.

Aged

Prototheca zopfii isolated from a patient with olecranon bursitis.

Prototheca zopfii was isolated from a patient with olecranon bursitis. Olecranon bursitis caused by Prototheca is a distinct clinical entity. Recognition of this infection was made by observing characteristic organisms in tissue and subsequent isolation of the organism.

Aged

Meningitis due to Prototheca wickerhamii in a patient with AIDS.

The first documented case of algal meningitis due to Prototheca wickerhamii is reported in a patient with AIDS. The initial CSF culture yielded only Cryptococcus neoformans. P. wickerhamii was isolated on four subsequent lumbar punctures. The patient died, and at autopsy the alga was isolated from leptomeninges over the brain and about the spinal cord. Histologic sections from numerous locations of the brain revealed masses of cryptococci and prototheca.

AIDS-Related Opportunistic Infections

[Fluconazole and itraconazole susceptibility testing with clinical yeast isolates and algae of the genus Prototheca by means of the Etest].

Preliminary own results suggest, that the Etest (produced by AB BIODISK, Solna, Sweden) performed on casitone medium meets the requirements of a routine test of yeast susceptibility to fluconazole and itraconazole. Testing of 46 clinical yeast isolates, of 5 strains of Exophiala dermatitidis and 4 strains of algae of the genus Prototheca revealed species-, genus- and strain-specific variations of the susceptibility to fluconazole and itraconazole. Candida glabrata was less susceptible to both triazoles than the other Candida species with exception of Candida krusei. Exophiala dermatitidis was highly susceptible to itraconazole. Prototheca wickerhamii and P. zopfii were resistant to both triazoles. Casitone medium is most appropriate for the determination of susceptibility to fluconazole and itraconazole by the Etest. The results of the Etest were comparable with those of a breakpoint test (microdilution method).

Antifungal Agents

Rapid identification of Prototheca species by the API 20C system.

The conventional auxanographic method of testing for the assimilation of carbohydrates and alcohols by the various species of Prototheca requires at least 2 weeks of incubation at 25 to 30 degrees C before definitive results are obtained. Even though Prototheca spp., in culture as well as in fixed tissues, can be identified more rapidly by fluorescent-antibody techniques in which species-specific reagents are used, such diagnostic facilities and reagents are not available in most diagnostic laboratories. The API 20C clinical yeast identification system, a commercially available ready-to-use micromethod, was found to permit the definitive identification of P. stagnora, P. wickerhamii, and P. zopfii within 4 days.

Carbohydrate Metabolism

Prototheca wickerhamii tenosynovitis.

Prototheca wickerhamii is an algae-like organism rarely implicated in skin or soft tissue infections. We describe the first case (to our knowledge) of Prototheca tenosynovitis which followed median nerve release for carpal tunnel syndrome. Clinical presentation, operative findings, histopathology, microbiology and treatment of this unusual infection are discussed.

Amphotericin B

First isolation of Prototheca species in Spain.

One-hundred forty samples of waste water were studied in search of algae of the genus Prototheca, that are potentially pathogenic to man. The samples were taken from collector outlets opening onto the Guadelquivir River. Nineteen cultures of Prototheca were isolated and identified as P. wickerhamii 9, P. zopffi 8 and P. stagnore 2. This is the first report of these algae in Spain and it is to be anticipated that these opportunist human pathogens will eventually be found to cause disease in the general population.

Chlorophyta

Growth of Prototheca isolates on n-hexadecane and mixed-hydrocarbon substrate.

Prototheca zopfii, an achlorphyllous alga, was capable of using hydrocarbons as sole carbon and energy source. The ability of P. zopfii to use hydrocarbons did not correlate with source of isolation. Seventy-five percent of the P. zopfii cultures recovered from sewage, plants, or animals utilized hydrocarbons. Other Prototheca species and P. zopfii that did not utilize hydrocarbons were isolated simultaneously from several sources with isolates that did use hydrocarbons. Species type rather than source of isolation was the predominant factor that determined hydrocarbon utilization.

Alkanes

Effect of growth rate on the macromolecular composition of Prototheca zopfii, a colorless alga which divides by multiple fission.

Prototheca zopfii, a eukaryote that divides by multiple fission, was investigated to determine how growth rate controls daughter cell number. The macromolecular composition, cell size, and number of nuclei per cell were determined in cultures during balanced growth in various media. Cellular mass, ribonucleic acid (RNA), deoxyribonucleic acid (DNA), carbohydrate, and nuclear number increased as positive linear functions of growth rate, whereas nuclear ploidy remained constant with a value of 0.098 pg of DNA/nucleus. The ratios of RNA to protein, protein to mass, and carbohydrate to mass were unaffected by growth rate, whereas the ratios of DNA to protein and RNA to DNA could be expressed as curvilinear functions of growth rate, the former negative and the latter positive. The dependency of normalized gene dosage (DNA/protein) on growth rate appeared as a distinguishing feature of multiple fission. Determination of the normalized rates of protein and RNA synthesis revealed that both increase linearly with growth rate. It is concluded that Prototheca zopfii may exist in a number of physiological states which are characterized by a unique size and macromolecular composition and which are dictated by growth rate.

Carbohydrates