Pitiose cutânea equina de localização atípica tratada topicamente com solução de anfotericina B e DMSO

Background: Cutaneous lesions by Pythium insidiosum infection are commonly observed in horses, especially in those living at flooded environments. Equine pythiosis is characterized by the development of tumoral masses that are frequently located at distal limbs, ventral abdomen, thorax, breast and face. The lesions are usually granulomatous, serosanguineous and ulcerated, most often destroyed by self-mutilation due to the intense pruritus. The proposed treatment includes surgical excision followed by antifungal drugs administration, which can be done systemically or topically. Amphotericin B and dimethyl sulfoxide (DMSO) in association has been successfully used for cutaneous pythiosis topical treatment due to the DMSO property to carry any substance through plasmatic membranes.Case: The present report concerns a 12-year-old mixed breed gelding presenting with self-mutilation of a tumoral mass located at the left flank. The owners reported that the horse had initially presented a small wound that had evolved to a 20-cm in diameter mass in 4 weeks. Tissue samples were collected, processed and stained by the Gomori's methenamine silver (GMS) method. The histopathological analysis revealed Pythium insidiosum hyphae in a granulomatous tissue, especially located at peripheral region, where kunkers were present. Surgical excision of the mass followed by cauterization was indicated as initial treatment, and due to financial reasons, the owners elected only the topical antifungal therapy to control the fungus infection after surgery. Flunixin meglumine was also administrated for five days aiming the control of pain and inflammation. The wound was cleaned with povidone-iodine solution and rinsed with a solution containing, 50 mg, of amphotericin B in 10 mL of sterile water and 10 mL of DMSO. This procedure was carried Out twice a day. The wound healed fast due to an excellent centripetal epithelialization. and the horse was discharged after 64 days showing only 5% of the initial wound area. The owner reported by telephone the complete healing and hair growth 10 days after discharge.Discussion: Despite the atypical location of the tumoral lesion described at the present report, the history and clinical manifestations, especially the intense pruritus, showed similarity with other characteristic reports of equine cutaneous pythiosis. The diagnosis was confirmed by the histopathological examination showing hyphae structures, as described to be evidences of the presence of Pythium insidiosum in the tissue. The surgical procedure was the first step to provide remission of clinical signs, and one day after surgery the pruritus desapeared. After excision of the granulomatous tissue and cauterization, daily topical administration of amphotericin B associated with DMSO was effective in destroying the infectious agent, as observed by the excellent epithelization. A pink granulation tissue grew up providing an ideal surface for epithelial migration and the healing process progressed quickly. Centripetal epithelialization reduced the wound area until 3% of the initial area in 64 days of treatment, when the remaining wound was found almost completely healed and covered with hair. At the present report, the horse presenting pythiosis was only topically treated. The recommended therapy using amphotericin B and DMSO solution was effective, economically viable and low risk, considering that the systemic antifungal therapy usually suggested is expensive and extremely nephrotoxic. The atypical location of the lesion on the left flank shows that any anatomical region can be affected by the fungus, since the conditions for its development were present.

Susceptibility of citrus species to Alternaria alternata, the causal agent of the Alternaria brown spot

Alternaria alternata, the causal agent of Alternaria brown spot (ABS), causes necrosis on leaves, twigs, and fruit, reducing the productivity and quality of fruits. Tangerines and their hybrids are highly susceptible to the disease. Species, hybrids, and cultivars of Citrus from the germplasm bank of the Estacao Experimental de Citricultura de Bebedouro, São Paulo, Brazil, were evaluated in 2004 and 2005 with respect to their resistance to A. alternata, both through natural infection and by inoculation. Detached leaves were also used to demonstrate susceptibility or resistance to the disease. Ten cultivars of Satsumas (Citrus unshiu), and 14 cultivars of Clementine mandarin (C. clementina) did not show any symptoms of the disease in their leaves, either through natural infection or when inoculated in the field. The Burguess SRA-412, Wallent SRA-438, Carvalhais, Ampefy SRA-459, Ananas SRA, and Macaque SRA-426 mandarin hybrids (C. reticulata) did not show symptoms of the disease under natural or artificial infection in the field. Some cultivars of C. deliciosa, C. tangerina, C. erythrosa, and C. temple showed symptoms of the disease, even though no previous record of their susceptibility to Alternaria brown spot had been previously reported. The hybrids Fairchild, Nova, Page, Fortune, and Sunburst were susceptible to the disease. However, Fremont mandarin (a crossing between C. clementina and C. reticulata), Encore (C. nobilis x C. deliciosa), and Fallglo (C. reticulata x C. paradisi) did not show symptoms in field, and few symptoms were verified in detached leaves. These materials are promising for the cultivation of tangerines, and will enable genetic improvement for the development of cultivars resistant to Alternaria brown spot. (c) 2007 Elsevier B.V. All rights reserved.

Biofertilizer for control of Guignardia citricarpa, the causal agent of citrus black spot

Biofertilizers generated from the anaerobic and aerobic digestion of cattle manure, known as Bio1 and Bio2, respectively, were studied with regard to their microbial composition and effect on the mycelial growth of Phyllosticta citricarpa, the causal agent of citrus black spot (CBS). Two field experiments were conducted to determine the biofertilizer's potential (Bio1) in controlling CBS (2001/2002 and 2002/2003 crops). It was observed that the greatest number of microorganisms was found in the aerobically produced biofertilizer. Bio2 did not inhibit the mycelial growth of P. citricarpa. Mycelial growth of P. citricarpa was inversely proportional to the Bio1 biofertilizer concentration. In the 2001/2002 cropping season, the Biol effect in controlling CBS was directly proportional to its concentration, at the rate of 0 (healthy fruit), with R-2 = 0.88. Biol had a significant effect in controlling CBS, at a concentration of 10%, during the 2001/2002 cropping season, with DI values of 0.246 and 0.229 for the. 10 and 20% doses, respectively, compared to DI of 0.329 for the control. A directly proportional effect of the biofertilizer concentration on the percentage of fruits with a rating of zero was. also observed in the 2002/2003 cropping season, with R-2 = 0.48. However, even at doses higher than in the preceding cropping season, the biofertilizer was less effective, possibly due to a higher occurrence of the disease. Copper oxychloride and combined applications of copper oxychloride and carbendazim plus mancozeb controlled the disease. The possibility of using the biofertilizer as a protective biofungicide to replace copper oxychloride, especially in organic agriculture, should be explored. (c) 2005 Elsevier Ltd. All rights reserved.

The predatory mite Stratiolaelaps scimitus as a control agent of the fungus gnat Bradysia matogrossensis in commercial production of the mushroom Agaricus bisporus

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

A new method for inoculation of fruit with Guignardia citricarpa, the causal agent of citrus black spot

Citrus black spot (CBS) is a fungal disease, caused by Guignardia citricarpa, that has a high economic impact on citrus. Although G. citricarpa has been associated with black spot of citrus, an adequate pathogenicity test is still not available. Thus, our objective was to develop and evaluate a simple, safe, and practical pathogenicity test. We used fruits from Pera-Rio and Valencia sweet orange trees from two different orchards, located in the State of São Paulo, Brazil. Inoculation was performed by placing six disks colonized by G. citricarpa, onto the peel of healthy fruits, previously bagged. In the Pera-Rio sweet orange grove, initial symptoms of the false melanose type resulting from the inoculations were observed 55 days after inoculation (dai). In the Valencia grove, initial symptoms also of the false melanose type resulting from the inoculations occurred 73 dai. A total of 92.8% and 86.6% of the Pera Rio and Valencia fruits inoculated, respectively, showed symptoms of CBS. Citrus black spot symptoms were not observed in any of the control fruits.

Genetic Diversity and Population Differentiation of the Causal Agent of Citrus Black Spot in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genetic diversity and antifungal susceptibility testing of Trichosporon asahii isolated of Intensive Care Units patients

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Potential tuberculostatic agent: Micelle-forming pyrazinamide prodrug

Pyrazinamide was condensed with the poly(ethylene glycol)-poly(aspartic acid) copolymer (PEG-PASP), a micelle-forming derivative was obtained that was characterized in terms of its critical micelle concentration (CMC) and micelle diameter. The CMC was found by observing the solubility of Sudan III in Poly(ethylene glycol)-poly(pyrazinamidomethyl aspartate) copolymer (PEG-PASP-PZA) solutions. The mean diameter of PEG-PASP-PZA micelles, obtained by analyzing the dynamic light-scattering data, was 78.2 nm. The PEG-PASP-PZA derivative, when assayed for anti-Mycobacterium activity, exhibited stronger activity than the simple drug.

Molecular typing and antifungal susceptibility of clinical sequential isolates of Cryptococcus neoformans from São Paulo State, Brazil

The antifungal susceptibility profiles and the genetic variability of 83 sequential clinical isolates of Cryptococcus neoformans, including four Cryptococcus gattii isolates, obtained from 38 São Paulo AIDS patients with cryptococcal meningitis were assessed by electrophoretic karyotyping and random amplified polymorphic DNA (RAPD) analysis. The majority of the Cryptococcus neoformans isolates were highly susceptible to amphotericin B and fluconazole. Twenty percent of the minimum inhibitory concentration values for amphotericin B varied from 0.5 to 1 mu g mL(-1). For fluconazole, 22% occurred in the range 8-16 mu g mL(-1). Sequential isolates from nine patients showed a trend towards lower susceptibility to fluconazole, flucytosine, itraconazole and amphotericin B. The results of molecular typing by electrophoretic karyotyping and RAPD analysis showed the presence of 22 electrophoretic karyotypes (EK) and 15 RAPD profiles that were highly correlated. Our results provided evidence for the occurrence of genetic changes in some strains associated with microevolution during the course of infection. We also observed both microevolution and simultaneous coinfection with two distinct Cryptococcus neoformans strains in one patient. In some patients, we found changed EK- and RAPD patterns in association with increased MIC values.

Amphotericin B Microemulsion Reduces Toxicity and Maintains the Efficacy as an Antifungal Product

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Spectrophotometric determination of ceftazidime in pharmaceutical preparations using neocuproin as a complexing agent

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Pothomorphe umbellata: Antifungal activity against strains of Trichophyton rubrum

Trichophyton rubrum is a dermatophyte, which can cause infections in human skin, hair and nail. Pothomorphe umbellata (L.) Miq. (Piperaceae) is a native Brazilian plant, in which phytochemical studies have demonstrated the presence of steroids, 4-nerolidylcatechol, sesquiterpenes and essential oils. The objective of this study was to analyze the in vitro activity of extracts and fractions of P. umbellata on resistant strains of T. rubrum. The microdilution plate method was utilized to test Tr1, H6 and Delta TruMDR2 strains of T rubrum; Delta TruMDR2 strain was obtained from H6 by TruMDR2 gene rupture, which is involved in multiple drugs resistance. The highest antifungal activity to all strains was observed for dichloromethane and hexane fractions of the 70% ethanolic extract which showed minimal inhibitory concentration (MIC) and minimal fungicide concentration (MFC) of 78.13 mu g/mL. This antifungal activity was also obtained by 70% ethanolic extract, which presented MIC and MFC of 78.13 mu g/mL to Delta TruMDR2, whereas the MIC values for Tr1 and H6 were 78.13 and 156.25 mu g/mL, respectively. Our results suggest the potential for future development of new antifungal drugs from P umbellata, especially to strains presenting multiple resistance. (C) 2012 Elsevier Masson SAS. All rights reserved.

Antitumor and immunomodulatory effects of the naphthoquinone 5-methoxy-3,4-dehydroxanthomegnin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)