Rabbits weighing 2 to 3 kg with induced Candida endophthalmitis in the left eye were given 5 mg/kg and 10 mg/kg pimaricin intravenously. In the course of investigation, no significant level of pimaricin was found in the vitreous of those animals receiving 5 to 10 mg/kg pimaricin intravenously, and only 3 microgram/ml aqueous was found after chronic therapy for 1 to 3 weeks. No ocular or retinotoxicity was noted, although toxicity was inherent in the liver, kidney, and endocrine glands. From these studies, it is concluded that intravenous therapy alone is not sufficient for treatment of fungal endophthalmitis due to high toxicity and inadequate ocular drug levels.

The Candida colonization and the dynamics of antibodies to Candida were studied in intensive care patients. 20 patients received no antifungal prophylaxis, 20 patients were treated by local application of Pimaricin into the mouth, nose, and into the respiratory tract during the time of intratracheal intubation. The frequencies and quantities of yeasts in the respiratory secretions were considerably reduced in the group receiving Pimaricin, whereas the amount of faecal colonization showed no difference between both groups of patients. During the 2nd-3rd week of hospitalization 17 patients of the untreated group and 4 patients of the treated group developed a significant rise of Candida antibodies. The association between the respiratory colonization and the antibody response indicates that the mucosa of the mouth and the respiratory tract is a main source of Candida invasion. In order to prevent that precondition of Candida endomycosis, routine ororespiratory antifungal prophylaxis is recommended, particularly in patients with prolonged intratracheal intubation.

Frequency of occurrence of strains of C. parapsilosis, their susceptibility to drugs and importance in pathogenesis of vaginal mycosis, was evaluated. Strains of C. parapsilosis were isolated in 44 cases which comprises 1.34% of a total number of 3275 tested Candida-like strains. Vaginal mycoses occurred in presence of Lactobacillus sp., normal pH values of vaginal contents, whereas leukocytosis was differing. Following drugs were used for testing susceptibility of C. parapsilosis: nystatin, pimaricin, amphotericin B, flucytosine, cotrimoxazole, miconazole, ketoconazole, tioconazole, fluconazole and itraconazole. It has been demonstrated that 38.3% strains of C. parapsilosis were exhibiting lowered activity toward amphotericin B, nystatin (23.5% resistant) and flucytosine (11.7% resistant). Tested strains were susceptible to remaining drugs within 94.1% and 100%.

A morphological interpretation is presented for data collected during growth of a filamentous organism, using a focused beam reflectance measurement (FBRM) system. The morphology of the organism was also obtained using conventional semiautomatic image analysis to support the interpretation of the FBRM data. The model organism employed is the filamentous soil-borne actinomycete Streptomyces natalensis, which produces the antifungal agent pimaricin. The organism was cultivated both in shake flasks and in a bench-scale stirred tank bioreactor. It was found that FBRM could be used to track changes in the morphology of the organism throughout the course of its growth on both scales. These changes were highlighted using both the median chord length and length-weighted mean chord length obtained from the chord length distribution measured with the FBRM probe. The ability of the FBRM probe to respond to changes in both the size and morphology of mycelial aggregates was supported by standard image analysis parameters, including equivalent diameter, convex area, and compactness.

The state of Michoacan is one of the main fresh pepper (Capsicum annuum L.) and tomato (Lycopersicon esculentum Mill.) producers in Mexico. During the last 5 years, pepper-producing areas in the state have become unproductive due to root-rotting pathogens, mainly Phytophthora capsici Leonian. Growers trying to overcome losses have increased tomato production in areas previously used for pepper production. Field-grown tomato plants with diseased green tomato fruits were observed in Tacambaro, Michoacan, during August 2002. Initially, brown-to-black lesions developed on fruits in contact with soil, followed by infection of the upper fruits in the raceme. Lesions enlarged and dark zonate "buckeye" bands were formed in the affected area. Diseased fruit turned mushy. Symptoms observed were similar to those described for buckeye rot of tomato (1). Diseased fruit were surface disinfested with 70% ethanol, cut into 0.5-cm slices, and incubated in a moist chamber to induce mycelial colonization. Isolation from mycelial tufts growing through the tomato slice was performed 3 days later, and mycelia was transferred to PARP selective medium (corn meal agar (CMA) plus ampicillin, pimaricin, rifampicin, and pentachloronitrobenzene). P. capsici was consistently isolated from diseased tomato fruits. Oomycete identification was based on sporangial and gametangial characteristics of cultures grown on CMA (1). Sporangia microscopically observed were spherical, broadly ellipsoid or obovoid with one papilla (occasionally two papillae), and deciduous with a long pedicel. Chlamydospores were not present (2). The isolates were heterothallic, and oogonia with amphigynous antheridia were observed in pairings with A1 and A2 isolates of P. capsici. Three isolates were A1 and two isolates were A2. To confirm pathogenicity, two experiments were performed using 20 healthy unwounded green tomatoes. One isolate of each mating type was tested. Isolates were grown for 5 days on CMA, and fruits were inoculated by placing P. capsici in contact with the fruit. Inoculated fruits were kept in a moist chamber at room temperature (17 to 20°C). Initial symptoms in the form of brown-to-black lesions appeared 24 h after inoculation. One week after inoculation, symptoms were similar to those observed in field-grown plants, and P. capsici was recovered from the margins of the diseased tissue. All inoculated fruits rotted. To our knowledge, this is the first report of P. capsici causing buckeye rot on tomato in Michoacan and of the presence of both mating types in the area. Reference: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society. St. Paul MN, 1996. (2) M. Aragaki and J. Y. Uchida. Mycologia 93:137, 2001.

Pigmented rabbits weighing 3 to 6 lbs were given bilateral intraocular injections of 250 to 1,000 mug pimaricin. Following their injection, blood, aqueous and vitreous levels were determined at various time intervals during the first 24 hours and at 24 hour intervals thereafter for one week. In subsequent studies, pigmented rabbits were given bilateral intraocular injections of 5,000 spores of A. fumigatus and 30 hours later received intraocular injections of 250 to 1,000 mug pimaricin. These studies show that 250 mug of intraocular pimaricin is well tolerated in the infected and normal animal eye with therapeutic ocular levels maintained for over 24 hours. Drug levels above 250 mug, although relatively notoxic in the normal eye resulted in irreversible damage to ocular structures in the infected eye that could not be resolved. Thus in the case of fungal endophthalmitis involving the anterior segment which will lead to the ultimate loss of the eye, an injection of 250 mug of pimaricin might preserve useful vision.

Frequency of appearance of Candida krusei in human reproductive tract, susceptibility of these microorganisms to drugs and their importance in pathogenesis of mycosis, were investigated. Strains C. krusei species constituted 6.7% of the total 1325 strains and 21.8% within other than C. albicans species yeast-like fungi. Vaginal mycosis caused by fungi from C. krusei species occurs in presence of Lactobacillus sp. and correct values of pH in vaginal contents, however changed leukocytosis in evident. Over 60% of patients with C. krusei infection were complaining that they feel disturbances within the reproductive tract. Investigation of susceptibility of C. krusei to drugs revealed highest activity of cotrimazine and pimaricin.

Evaluation of 25, 50, and 100 mug of pimaricin given intravitreally to albino rabbits with Aspergillus endophthalmitis revealed that 25 mug of pimaricin, while nontoxic to the retina, was ineffective in inhibiting the fungal process. A dose of 50 mug was significantly more effective in inhibiting fungal disease but also caused significant retinal damage with loss of retinal function and iridoplegia. Dosages above this level resulted in vitreous retraction and degeneration, iridoplegia, and retinal detachment. Twenty-five micrograms of pimaricin given in three dosages spaced three days apart may be effective in inhibiting fungal endophthalmitis.

The action of the polyene macrolide antibiotics mycotrienin, pimaricin, lucensomycin, tetramycin, rimocidin, nystatin, filipin, lagosin, pentaene antibiotic 2814P, flavomycoin, flavofungin, hexaene antibiotic 5001P, and candicidin, including perhydro derivatives of them, on wall-less stable protoplast type L-form and normal rod form cells of E. coli W1655F+ was studied. No inhibition of the normal rod form cells was detected. In contrast to these results the growth of the L-form cells was inhibited by all of the substances tested, with the exception of pimaricin. Further experiments have shown that the differences in sensitivity of normal and L-form cells cannot be explained by differences in sterol content, the target site of polyene antibiotics in sensitive eukaryotic cells. According to our results it is obvious that the cell wall of the normal cells functions as a penetration barrier to polyene antibiotics.