Publications by authors named "Marlete Brum Cleff"

26 Publications

  • Page 1 of 1

Multiresistant bacteria isolated from domestic and wild animals with skin lesions were susceptible to native plants from Southern Brazil.

Nat Prod Res 2021 Jun 2:1-5. Epub 2021 Jun 2.

Department of Veterinary Clinic, Faculty of Veterinary, Federal University of Pelotas, Pelotas, RS, Brazil.

We evaluated the chemical composition, toxicity, and antibacterial activity of (SCH), (EUG), (PER), (EQU), (SOL), and (BRA). These plants were tested (7.5-0.01 mg/mL) against Gram-positive (G+;  = 32) and Gram-negative (G-;  = 26) isolates from animals (M07-A9, CLSI). Antibiogram (disk diffusion), chromatographic analysis (UPLC), and toxicity assay (HET-CAM) were also performed. A high incidence of resistance was noted, in which 18.4% (07/38) of G+ () and 17.7% (06/34) of G- () were multidrug-resistant. All bacteria were sensitive (MIC) to SCH (both 3.75 mg/mL), EUG (3.75 mg/mL and 7.5 mg/mL, respectively) and PER (both 7.5 mg/mL). SCH/EUG/PER highlighted as antibacterial, probably due to the major compounds (ethyl gallate, quinic acid, quercetin). These extracts showed normal embryonic development (SCH/EUG: 7.5-0.94 mg/mL). These findings highlighted the promising use of native plants for therapeutic purposes.
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http://dx.doi.org/10.1080/14786419.2021.1933972DOI Listing
June 2021

Dioctophimosis: A Parasitic Zoonosis of Public Health Importance.

Adv Exp Med Biol 2021 ;1306:129-142

Project Dioctophyme renale in Dogs and Cats (PRODIC), Department of Veterinary Clinics, Veterinary College, Federal University of Pelotas, Pelotas, RS, Brazil.

Dioctophyme renale, the giant kidney worm, is a renal nematode from domestic and wild mammals that has zoonotic potential. In humans, dioctophimosis has been reported in several countries, mainly on the Asian continent, totaling more than 40 cases, which describe the parasite mainly infecting the kidneys, bladder, urethra and skin. Infection in animals and humans is related to the ingestion of the infective larva (L3) present in the aquatic oligochaete annelid (mandatory intermediate host) or fish and anurans (facultative paratenic hosts). Thus, the infection is related to the habit of drinking water contaminated with the mandatory intermediate host, as well as raw or undercooked meat from the facultative paratenic hosts. Dioctophimosis destroys the renal parenchyma and, in some cases, can cause the death of its hosts. In this chapter, we discuss the main topics regarding dioctophimosis in humans, domestic and wild animals, highlighting its importance in public health.
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http://dx.doi.org/10.1007/978-3-030-63908-2_10DOI Listing
May 2021

Disseminated histoplasmosis in a cat rescued in Fortaleza, Brazil, and successfully treated with itraconazole - First case report identified molecularly.

Med Mycol Case Rep 2020 Dec 2;30:29-34. Epub 2020 Oct 2.

Faculty of Veterinary, Federal University of Pelotas, Rua Campus Universitario, 1, Pelotas/RS, 96010-900, Brazil.

An unneutered female cat of unknown age presented bloody lesions on the edematous face, and respiratory signs. Cytology and culture from the skin sample collected with fine-needle aspiration showed yeasts inside activated macrophages, and fungal growth characteristic of spp., which was molecularly confirmed that was var. capsulatum. The cat was successfully treated with oral itraconazole (10 mg/kg/daily) for 120 days. This is the first case report of feline histoplasmosis confirmed molecularly in Brazil.
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http://dx.doi.org/10.1016/j.mmcr.2020.09.005DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7559507PMC
December 2020

Jabuticaba [ (Poir.) Govaerts]: a Brazilian fruit with a promising application against itraconazole-susceptible and -resistant .

Nat Prod Res 2020 Aug 25:1-5. Epub 2020 Aug 25.

Department of Preventive Veterinary, Faculty of Veterinary, Federal University of Pelotas, Pelotas, RS, Brazil.

Hydroalcoholic extract from Jabuticaba peels was evaluated for the chemical profile, antioxidant potential, cytotoxicity, and anti- activities against both dimorphic phases. Higher phenolic content (14.91 ± 0.97) compared to flavonoid (2.05 ± 1.00) associated with the ellagic acid (1.93 ± 0.03; LC-MS), and a good scavenging ability against ABST and DPPH radicals was noted. On MDBK cells, elevated cell viability (>90%) was demonstrated between 0.39 and 0.097 mg/ml (MTT assay). Mycelial (CLSI M38-A2) and yeast (CLSI M27-A3) phases of 18 isolates of from cats ( = 8), dogs ( = 8) and humans ( = 2) were used. They were identified itraconazole-susceptible and itraconazole-resistant isolates in both phases, which were all inhibited (MIC of ≤1.56-6.25 mg/ml for both phases) and killed (MFC of ≤1.56-12.5 mg/ml for mycelial; ≤1.56-50 mg/ml for yeast) by Jabuticaba. For the first time, these findings highlighted the potential usefulness of hydroalcoholic extract from Jabuticaba peel as a promising antifungal against sporotrichosis.
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http://dx.doi.org/10.1080/14786419.2020.1810034DOI Listing
August 2020

Benzylidene-carbonyl compounds are active against itraconazole-susceptible and itraconazole-resistant Sporothrix brasiliensis.

Folia Microbiol (Praha) 2020 Dec 21;65(6):1033-1038. Epub 2020 Aug 21.

Department of Pharmaceutical Sciences, Institute of Environmental, Chemical and Pharmaceutical Sciences, Federal University of São Paulo (UNIFESP), Diadema, SP, 09913-030, Brazil.

We evaluated the antifungal activity of benzylidene-carbonyl compounds (LINS03) based on the structure of gibbilimbol from Piper malacophyllum Linn. Five analogues (1-5) were synthetized following a classic aldol condensation between an aromatic aldehyde and a ketone, under basic conditions. These were tested against itraconazole-susceptible (n = 3) and itraconazole-resistant (n = 5) isolates of Sporothrix brasiliensis by M38-A2 guidelines of CLSI. All of them were fungistatic (MIC ranged of 0.11-0.22 mg/mL (1); 0.08-0.17 mg/mL (2); 0.05-0.1 mg/mL (3); 0.04-0.33 mg/mL (4); and 0.04-0.3 mg/mL (5)), highlighting compounds 2 and 3. As fungicidal, compounds 1 and 2 were highlighted (MFC ranged of 0.22-0.89 mg/mL and 0.08-1.35 mg/mL, respectively), compared with the remaining (0.77-> 3.08 mg/mL (3); 0.08-> 2.6 mg/mL (4); and 0.59-> 2.37 mg/mL (5)). The inhibitory activity was related to the benzylidene-carbonyl, whereas the phenol group and the low chain homolog seems to contribute to some extent to the fungicidal effect. Compound 2 highlighted due to the considerable fungistatic and fungicidal activities, including itraconazole-resistant Sporothrix brasiliensis. These findings support the potential usefulness of benzylidene-carbonyl compounds as promising prototypes for the development of antifungal against sporotrichosis by Sporothrix brasiliensis, including against itraconazole-resistant isolates.
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http://dx.doi.org/10.1007/s12223-020-00814-4DOI Listing
December 2020

Feline sporotrichosis: a case series of itraconazole-resistant Sporothrix brasiliensis infection.

Braz J Microbiol 2021 Mar 9;52(1):163-171. Epub 2020 May 9.

Department of Veterinary Clinic, Faculty of Veterinary, Federal University of Pelotas, 1 Campus Universitário Capão do Leão, Pelotas/RS, Rio Grande do Sul, 96010-900, Brazil.

The treatment of feline sporotrichosis is a challenge for veterinary clinicians since refractory cases may occur, due either to patient and/or to pharmacological management errors or due to the development of antifungal resistance. Thus, we aimed to describe the therapeutic history of feline cases infected by itraconazole-resistant Sporothrix brasiliensis in an endemic region of Southern Brazil. Medical records of cats attended at the Veterinary Clinic Hospital (Pelotas/RS, Brazil) between 2016 and 2017 were reviewed. Twelve cases of infection by S. brasiliensis with that showed high minimum inhibitory concentration (MIC) values (≥ 4 μg/mL) to itraconazole by M38-A2 of CLSI were selected. At the hospital consultation, disseminated (cats 1-l0, 12) and localized (cat 11) skin lesions remained in the cats, even after treatment with fluconazole, ketoconazole (02/12), and itraconazole (ITZ, 09/12) performed before this study. High doses (25-100 mg/kg/day) of ITZ for up to 4 months (03/12, cats 2, 6, 12) or over 12 months (05/12, cats 1, 5, 7, 8, 11) did not provide a clinical cure, except for the association of ITZ plus potassium iodide (01/12, cat 12) for 3 months, which proved useful in infections with itraconazole-resistant S. brasiliensis. However, the combined issues of abandonment of therapy by owners for financial reasons, difficulties surrounding therapy administration (03/12, cats 6, 11, 12), and the inappropriate choice of medication (01/12, cat 6), together reflect the reality of this endemic region, which greatly compromises clinical healing. This study highlighted the occurrence of refractory cases by itraconazole-resistant S. brasiliensis in cats from Southern Brazil, as well as the abandonment of treatment and therapeutic errors. We warn of the need for antifungal susceptibility tests to adapt therapeutic protocols in feline sporotrichosis.
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http://dx.doi.org/10.1007/s42770-020-00290-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7966689PMC
March 2021

Susceptibility and resistance of Sporothrix brasiliensis to branded and compounded itraconazole formulations.

Braz J Microbiol 2021 Mar 24;52(1):155-162. Epub 2020 Apr 24.

Departamento de Veterinária Preventiva, Faculdade de Veterinária, Universidade Federal de Pelotas, Pelotas, Rio Grande do Sul, Brazil.

Itraconazole is the first drug of choice for the treatment of sporotrichosis and it is available at different concentrations for veterinary patients. However, therapeutic failure has been reported, limiting clinical treatment. This study evaluated the in vitro efficacy of brand-name and compounded itraconazole formulations against Sporothrix brasiliensis and estimated the itraconazole content in each tested formulation. Oral capsules were acquired from two brand-name products for human (H-IND) and veterinary (V-IND) uses, and three from compounding pharmacies in Pelotas, RS, for human (H-COMP1/H-COMP2) and veterinary (V-COMP) uses. Capsule purity was analyzed by liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS). Antifungal activity was determined against 29 Sporothrix brasiliensis by the M38-A2 guideline of CLSI. H-IND/H-COMP1/H-COMP2 had high efficacy against S. brasiliensis (approximately 70% of total isolated susceptible), V-COMP showed moderate efficacy (51.7%), and V-IND was the least effective formulation (37.9%). Thirty-four percent of the total isolates were resistant to all formulations. Furthermore, itraconazole content did not match the concentration indicated by the manufacturers, ranging from 387.70 to 7.81 μg/mg (H-COMP2 > V-COMP > H-IND > H-COMP1 > V-IND). Therefore, it is possible that the formulations showed different in vitro efficacy due to the difference in their itraconazole contents. Given the emergence of antifungal resistance for all formulations, the choice product to be used must follow susceptibility testing. Stringent quality control measures are recommended for product manufactures to assure drug content uniformity.
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http://dx.doi.org/10.1007/s42770-020-00280-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7966699PMC
March 2021

protection of the marjoram ( Linn.) essential oil in the cutaneous sporotrichosis by .

Nat Prod Res 2019 Oct 17:1-5. Epub 2019 Oct 17.

Departamento de Veterinária Preventiva, Universidade Federal de Pelotas , Pelotas , Brasil.

Thirty Wistar rats subcutaneously infected by an itraconazole-resistant received the oral daily treatment ( = 10, each) of control (CTL, saline solution), itraconazole (ITZ, 10 mg/kg) and marjoram essential oil (MRJ, 80 mg/kg) for 30 days. Weekly, the clinical evaluation and euthanasia for histopathology and fungal burden were performed. Only animals from MRJ evolved to the remission of the cutaneous lesion with a mild to absent presence of yeasts in footpad, besides decreased the fungal burden in the systemic organs compared to CTL and ITZ ( < 0.05), preventing the fungal spread, mainly in the liver and spleen. The antifungal activity may have been attributed to the majority composition of terpinen-4-ol (34.09%), γ-terpinene (14.28%) and α-terpinene (9.6%), which the mode of action was at the level of ergosterol complexation. These findings highlighted the antifungal and the systemic protective effects of MRJ, supporting the promising use in the treatment of cutaneous sporotrichosis.
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http://dx.doi.org/10.1080/14786419.2019.1678617DOI Listing
October 2019

Ovicidal and larvicidal potential of Rosmarinus officinalis to control gastrointestinal nematodes of sheep.

Rev Bras Parasitol Vet 2019 Oct-Dec;28(4):807-811

Laboratório de Microbiologia, Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas - UFPel, Pelotas, RS, Brasil.

Gastrointestinal Nematode Infection (GIN) are the main constraint to the production of small ruminants. Studies of medicinal plants have been an important alternative in the effort to control these parasites. Therefore, the purpose of this study was to evaluate the in vitro ovicidal and larvicidal activity of essential oil of Rosmarinus officinalis. The oil was extracted, analyzed by gas chromatography and tested on GIN eggs and larvae in six concentrations, 227.5mg/mL, 113.7mg/mL, 56.8mg/mL, 28.4mg/mL, 14.2mg/mL and 7.1mg/mL. To determine the ovicidal activity, GIN eggs were recovered from sheep feces and incubated for 48h with different concentrations of the oil. For the evaluation of larval migration, third-stage larvae (L3) were obtained by fecal culture, and associated with the essential oil for 24h at the same concentrations, after which they were left for another 24 hours on microsieves, followed by the count of migrating and non-migrating larvae. The assays of R. officinalis oil showed a significant (p<0.05) 97.4% to 100% inhibition of egg hatching and a significant (p<0.05) 20% to 74% inhibition of larval migration. The main constituent revealed by gas chromatography was Eucalyptol. The results indicate that R. officinalis essential oil has ovicidal and larvicidal activity on sheep GINs.
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http://dx.doi.org/10.1590/S1984-29612019060DOI Listing
January 2020

Chemical composition and cytotoxicity of extracts of marjoram and rosemary and their activity against Sporothrix brasiliensis.

J Med Microbiol 2017 Jul 12;66(7):1076-1083. Epub 2017 Jul 12.

Department of Pharmacology, Institute of Basic Sciences and Health, Federal University of Rio Grande do Sul, Porto Alegre/RS, Brazil.

Purpose: Motivated by increasing reports of antifungal resistance in human and animal sporotrichosis, this study evaluated the chemical composition, cytotoxicity and anti-Sporothrix brasiliensis activity of extracts of marjoram (Origanum majorana) and rosemary (Rosmarinus officinalis).

Methodology: Ten (INF10) and 60 min (INF60) infusions, a decoction and a hydroalcoholic extract (HAE, 70 %) were prepared from both plants (10 % w/v). The extract composition was analysed by liquid chromatography/mass spectrometry and the cytotoxicity was evaluated using a colorimetric assay in canine and feline kidney cells. Using a broth microdilution assay (CLSI M38-A2) adapted to the extracts, 30 Sporothrix brasiliensis isolates from dogs, cats and humans, and one Sporothrix schenckii were tested.Results/Key findings. The predominant phenolic compounds found in all extracts were 4-hydroxybenzoic acid, caffeic acid and chlorogenic acid. Luteolin was also one of the predominant compounds, but only in the HAE of marjoram. Extracts of marjoram maintained cell viability in concentrations up to 2.5 mg ml-1 for the feline cell line and up to 10 mg ml-1 for the canine cell line, whereas in rosemary, the cell viability for both kidney lines was maintained with concentrations up to 5 mg ml-1. The activity of rosemary extracts was low or absent. Among the marjoram extracts, HAE was highlighted and had fungistatic activity against Sporothrix brasiliensis (MIC5040 mg ml-1), including in all itraconazole-resistant isolates. S. schenckiisensu stricto was sensitive to marjoram extracts (MIC/MFC ≤5 mg ml-1), with the exception of INF10.

Conclusion: These findings support the potential usefulness of the HAE of marjoram in the treatment of sporotrichosis.
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http://dx.doi.org/10.1099/jmm.0.000517DOI Listing
July 2017

Polar Origanum vulgare (Lamiaceae) extracts with antifungal potential against Sporothrix brasiliensis.

Med Mycol 2018 Feb;56(2):225-233

Department of Preventive Veterinary, Federal University of Pelotas, Pelotas, RS, Brazil.

Oregano (Origanum vulgare) has anti-Sporothrix spp. activity, including against strains that are resistant to antifungal drugs. As the studies are limited to the essential oil, the aim of this study was to evaluate the chemical, antioxidant and cytotoxic properties of polar oregano extracts and their anti-Sporothrix brasiliensis activity. Aerial plant parts were used in the preparation of 10 min (INF10) and 60 min (INF60) infusions, a decoction (DEC) and a hydroalcoholic extract (HAE). Six phenolic acids and four flavonoids were identified and quantified through liquid-chromatography (LC-MS). Extracts in increasing order of total phenolic and flavonoid contents were HAE 40 mg/ml for the other extracts. This is the first report of oregano extracts showing antifungal activity against S. brasiliensis. Its use in the treatment of sporotrichosis may be considered upon toxicity and safe-use conditions are tested.
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http://dx.doi.org/10.1093/mmy/myx031DOI Listing
February 2018

Plants from Lamiaceae family as source of antifungal molecules in humane and veterinary medicine.

Microb Pathog 2017 Mar 26;104:232-237. Epub 2017 Jan 26.

Centro de Diagnóstico e Pesquisa em Micologia Veterinária, Departamento de Veterinária Preventiva, Faculdade de Veterinária, Universidade Federal de Pelotas (UFPEL), Pelotas, RS, Brazil.

This work aimed to review the main plants of Lamiaceae family with activity against pathogenic fungi of medical and veterinary interest. Published studies in the main international databases between January 2002 and June 2016 showed that 55 botanical species belonging to 27 genus presented antifungal activity in different forms of extractions, mainly essential oils. Pathogenic fungi of Aspergillus spp., Candida spp., Malassezia spp., Cryptococcus spp., Sporothrix spp., Microsporum spp., Trichophyton spp. and Epidermophyton spp. genus were in vitro sensitive to several plants of Lamiaceae family. Chemical molecules isolated were described as promising use as antifungals in mycoses, highlighting estragole, 1,8-cineole, terpineol-4, γ-terpinene, among others. However, it should be alert to need of universal standardization in the laboratories tests with natural products.
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http://dx.doi.org/10.1016/j.micpath.2017.01.050DOI Listing
March 2017

Dioctophyme renale (Nematoda: Enoplida) in domestic dogs and cats in the extreme south of Brazil.

Rev Bras Parasitol Vet 2017 Jan-Mar;26(1):119-121. Epub 2016 Dec 1.

Clínica Cirúrgica Veterinária, Departamento de Clínicas Veterinária, Faculdade de Veterinária, Universidade Federal de Pelotas - UFPel, Pelotas, RS, Brasil.

Dioctophyme renale is a zoonotic nematode that parasites the kidneys of wild and domestic carnivores, and it has been reported frequently in Brazil. The aim here was to register the number of cases of dogs and cats diagnosed with dioctophymosis by necropsy (1981 to 2014) and ultrasound examination (2010 to 2015) in Pelotas-RS. In this context, a survey was conducted on dioctophymosis cases diagnosed at the Veterinary Pathology Laboratory (LPV) and Veterinary Clinical Hospital (HCV) of the Federal University of Pelotas (UFPel), and at a specialist veterinary imaging diagnostics clinic. In total, 95 cases were registered. The high series of the disease in dogs can be related to the presence of a large number of stray and semi-domestic dogs in the city, and also due to the ingestion of intermediate hosts of D. renale parasitized with the infective larvae. Thus, it can be concluded that Pelotas is a city with favorable conditions for the occurrence of dioctophymosis with high rate of disease in recent years.
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http://dx.doi.org/10.1590/S1984-29612016072DOI Listing
May 2018

First isolation of Dioctophyme renale eggs from an urban environment and identification of those from animal urine.

Rev Bras Parasitol Vet 2017 Jan-Mar;26(1):89-91. Epub 2016 Dec 1.

Faculdade de Veterinária, Universidade Federal de Pelotas - UFPel, Pelotas, RS, Brasil.

Dioctophyme renale is a zoonotic parasite with worldwide distribution, although its occurrence is little known. The objective here was to evaluate the presence of parasite eggs in the environment and in the urine of dogs and cats in an urban area. Soil samples and urine were evaluated respectively by means of the Caldwell-Caldwell technique and urinalysis. Out of the 100 soil samples, 3% presented D. renale eggs, and out of the 43 urine samples, 18.6% were positive, including the feline samples. Thus, D. renale eggs are present in the urban environment, and dogs and cats are parasitized by this nematode, which therefore represents a risk to public health.
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http://dx.doi.org/10.1590/S1984-29612016064DOI Listing
May 2018

In Vitro Susceptibility of Sporothrix brasiliensis to Essential Oils of Lamiaceae Family.

Mycopathologia 2016 Dec 16;181(11-12):857-863. Epub 2016 Aug 16.

Departamento de Farmacologia, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil.

This study evaluated the chemical, cytotoxic and anti-Sporothrix brasiliensis properties of commercial essential oils of rosemary (Rosmarinus officinalis L.), oregano (Origanum vulgare L.) and marjoram (Origanum majorana L.). Chemical composition of the oils was identified through gas chromatography with flame ionization detector, and cytotoxicity was performed through MTT assay in VERO cell line. Anti-S. brasiliensis activity was performed according to the CLSI M38-A2 guidelines using isolates obtained from cats and dogs. The major compounds found were carvacrol in the oregano oil (73.9 %) and 1,8-cineole in rosemary and marjoram oils (49.4 and 20.9 %, respectively). All S. brasiliensis isolates were susceptible to the plant oils, including itraconazole-resistant ones. Marjoram and rosemary oils showed MIC of 0.56 and 1.12 mg ml, and MFC of 4.5 and 9 mg ml, respectively. For oregano oil, a strong antifungal activity was observed with MIC and MFC values ≤0.07 mg ml. The weakest cytotoxicity was observed for rosemary oil. Further studies should be undertaken to evaluate the safety and efficacy of these essential oils in sporotrichosis.
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http://dx.doi.org/10.1007/s11046-016-0047-yDOI Listing
December 2016

Cytotoxicity and anti-Sporothrix brasiliensis activity of the Origanum majorana Linn. oil.

Braz J Microbiol 2016 Oct - Dec;47(4):896-901. Epub 2016 Jul 29.

Universidade Federal do Rio Grande do Sul (UFRGS), Instituto de Ciências Básicas da Saúde, Departamento de Farmacologia, Porto Alegre, RS, Brazil.

The study aimed to evaluate the anti-Sporothrix sp. activity of the essential oil of Origanum majorana Linn. (marjoram), its chemical analysis, and its cytotoxic activity. A total of 18 fungal isolates of Sporothrix brasiliensis (n: 17) from humans, dogs and cats, and a standard strain of Sporothrix schenckii (n: 1) were tested using the broth microdilution technique (Clinical and Laboratory Standard Institute - CLSI M27-A3) and the results were expressed in minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC). The MIC and MIC of itraconazole against S. brasiliensis were 2μg/mL and 8μg/mL, respectively, and the MFC and MFC were 2μg/mL and >16μg/mL, respectively, with three S. brasiliensis isolates resistant to antifungal. S. schenckii was sensitive at MIC of 1μg/mL and MFC of 8μg/mL. For the oil of O. majorana L., all isolates were susceptible to MIC of ≤2.25-9mg/mL and MFC of ≤2.25-18mg/mL. The MIC and MIC were ≤2.25mg/mL and 4.5mg/mL, respectively, and the MFC values were twice more than the MIC. Twenty-two compounds were identified by gas chromatography with a flame ionization detector (CG-FID) and 1,8-cineole and 4-terpineol were the majority. Through the colorimetric (MTT) assay, the toxicity was observed in 70-80% of VERO cells between 0.078 and 5mg/mL. For the first time, the study demonstrated the satisfactory in vitro anti-Sporothrix sp. activity of marjoram oil and further studies are needed to ensure its safe and effective use.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5052358PMC
http://dx.doi.org/10.1016/j.bjm.2016.07.017DOI Listing
January 2017

The use of (1-3) β-glucan along with itraconazole against canine refractory sporotrichosis.

Mycopathologia 2014 Apr 21;177(3-4):217-21. Epub 2014 Mar 21.

Mestranda em Clínicas Veterinária, Universidade Federal de Pelotas, Capão do Leão, RS, Brazil,

Sporotrichosis, caused by the Sporothrix schenckii fungal complex, is a zoonotic mycosis distributed worldwide. Itraconazole is the treatment of choice for domestic animals although some fungal isolates have shown resistance to this drug. The objective of this study was to report, for the first time, the use of (1-3) β-glucan along with itraconazole in the treatment of a canine with sporotrichosis caused by Sporothrix brasiliensis. The animal had ulcerated and crusted lesions, especially on the nasal planum. Clinical samples were collected for a complete blood count, cytological analysis of the lesion, and fungal culture. Based on the results of the laboratory examination, and after the fungal culture, antibiotic therapy and treatment with itraconazole were initiated. Two additional fungal cultures were performed, which were positive. After 7 months of the animal treatment with itraconazole, the S. brasiliensis culture was still positive, so that the itraconazole was associated with (1-3) β-glucan. After four weekly applications of glucan, the complete elimination of the fungus was observed based on the fungal culture negative results. The results show, therefore, that (1-3) β-glucan with itraconazole promoted the case resolution, and it may be considered a promising alternative for the treatment of sporotrichosis in cases of resistance to conventional therapy.
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http://dx.doi.org/10.1007/s11046-014-9736-6DOI Listing
April 2014

Clinical and mycological analysis of dog's oral cavity.

Braz J Microbiol 2013 9;44(1):139-43. Epub 2013 Apr 9.

Centro de Diagnóstico e Pesquisa em Micologia Veterinária, Departamento de Veterinária Preventiva, Universidade Federal de Pelotas, Pelotas, RS, Brazil.

The oral microbiota of humans and animals is made up of a wide variety of yeasts and bacteria, but microbiota of dogs is not totally described. Although such identification is an important step to establish the etiopathogenesis and adequate therapy for the periodontal disease The aim of this study was to evaluate and correlate oral alterations with the presence of yeasts in oral cavity of female dogs. After clinical evaluation samples from healthy and from dogs with oral diseases were obtained from three different oral sites by swabs, curettes, millimeter periodontal probes and HA membrane tip in cellulose ester. Yeast identification was performed through macroscopic and microscopic colony features and biochemical tests. Dental calculus was the most prevalent occurrence in the oral cavity of 59 females. However, the isolation of yeasts was significantly higher (p < 0.05) in animals suffering from halitosis. Eleven yeast species were identified, namely: Malassezia pachydermatis, Rhodotorula spp., Candida albicans, C. catenulata, C. famata, C. guilliermondii, C. parapsilosis, C. intermedia, Trichosporon asahii, T. mucoides and Cryptococcus albidus. It could be concluded that the yeasts are part of the microbiota from the different sites of the oral cavity of the female canines studied without causing any significant alterations except halitosis.
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http://dx.doi.org/10.1590/S1517-83822013005000018DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3804190PMC
October 2013

Inhibitory effect of sodium hypochlorite and chlorhexidine digluconate in clinical isolates of Sporothrix schenckii.

Mycoses 2012 May 8;55(3):281-5. Epub 2011 Sep 8.

Laboratory of Infectious Diseases, Federal University of Pelotas, Brazil Federal University of Rio Grande do Sul, Brazil.

The susceptibility of Sporothrix schenckii isolates from clinical cases of canine, feline and human sporotrichosis, and from the environment, was evaluated with 4% sodium hypochlorite and 6.6% chlorhexidine digluconate using the broth microdilution, agar diffusion and direct exposure techniques. The minimal inhibitory concentration was smaller than 0.8% for chlorhexidine digluconate and between 8% and 4% for sodium hypochlorite. Inhibition zones were not found in agar diffusion for sodium hypochlorite, and zones averaging 1.9 mm were found for chlorhexidine digluconate. In the direct exposure test, sodium hypochlorite demonstrated best performance at 20 min of contact, as chlorhexidine digluconate presented little antimicrobial activity.
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http://dx.doi.org/10.1111/j.1439-0507.2011.02101.xDOI Listing
May 2012

[Occurrence of Cryptococcus neoformans in pigeon excrement in the city of Pelotas, State of Rio Grande do Sul].

Rev Soc Bras Med Trop 2010 Mar-Apr;43(2):198-200

Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS.

Introduction: Cryptococcus neoformans is an encapsulated yeast and is the etiological agent for human and animal cryptococcosis. It is found in sources within the environment, including pigeon excrement, and is an important cause of mortality among immunocompromised individuals worldwide.

Methods: Seventy different environments in the city of Pelotas, Rio Grande do Sul, were surveyed for the purpose of investigating Cryptococcus neoformans occurrences in pigeon excreta. The environments included buildings, public squares and outdoor locations in the city. After collection, chloramphenicol saline solution was added to the excreta, which were then homogenized and seeded onto Sabouraud agar with chloramphenicol and onto Niger agar, and incubated at 32 degrees C. Identification was performed by direct examination and by means of the phenoloxidase and urease tests, carbohydrate assimilation and culturing in CGB medium.

Results: Out of the sites investigated (n = 70), 26 (37.1%) of them contained pigeon excreta. These included historical buildings (n = 8), a church tower (n = 1), rice mills and warehouses (n = 7), a public square (n = 1) and outdoor locations (n = 9). Cryptococcus neoformans was isolated from 26.9% (n = 7/26) of these locations.

Conclusions: This study draws attention to isolation of this fungus in urban areas that present large accumulations of pigeon excrement. This represents a risk to public health, especially for immunocompromised individuals.
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http://dx.doi.org/10.1590/s0037-86822010000200018DOI Listing
August 2010

[Experimental cutaneous sporotrichosis: in vivo evaluation of itraconazole and terbinafine].

Rev Soc Bras Med Trop 2009 Nov-Dec;42(6):706-10

Pós-Graduação em Veterinária, Faculdade de Veterinária, Universidade Federal de Pelotas, Pelotas, RS.

The aim of this study was to evaluate the in vivo activity of itraconazole and terbinafine for treating experimental cutaneous sporotrichosis. Eighty Wistar rats were used, divided into four groups (TERB20, TERB30, ITRA and CONT). They were inoculated in the left plantar pad with 0.2 ml of solution containing 2x10(3) cell/ml of Sporothrix schenckii and treated with terbinafine (20 and 30 mg/kg), itraconazole (10 mg/kg) or placebo for 13 weeks. The lesions at the inoculation site were evaluated and measured weekly, along with their dissemination. Mycological and histopathological analyses were performed subsequently. The results showed that the animals in the ITRA group differed statistically in all parameters evaluated, in relation to CONT. For terbinafine, there were no statistical differences between the treated and control groups. It could be confirmed that itraconazole presented good activity for treating sporotrichosis, while terbinafine was ineffective for this disease at the doses used. However, more studies on the latter antifungal agent are needed.
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http://dx.doi.org/10.1590/s0037-86822009000600018DOI Listing
May 2010

In vitro activity of origanum vulgare essential oil against candida species.

Braz J Microbiol 2010 Jan 1;41(1):116-23. Epub 2010 Mar 1.

Programa de Pós-graduação em Ciências Veterinária, Faculdade Veterinária, Universidade Federal do Rio Grande do Sul , Porto Alegre, RS , Brasil.

The aim of this study was to evaluate the in vitro activity of the essential oil extracted from Origanum vulgare against sixteen Candida species isolates. Standard strains tested comprised C. albicans (ATCC strains 44858, 4053, 18804 and 3691), C. parapsilosis (ATCC 22019), C. krusei (ATCC 34135), C. lusitaniae (ATCC 34449) and C. dubliniensis (ATCC MY646). Six Candida albicans isolates from the vaginal mucous membrane of female dogs, one isolate from the cutaneous tegument of a dog and one isolate of a capuchin monkey were tested in parallel. A broth microdilution technique (CLSI) was used, and the inoculum concentration was adjusted to 5 x 10(6) CFU mL(-1). The essential oil was obtained by hydrodistillation in a Clevenger apparatus and analyzed by gas chromatography. Susceptibility was expressed as Minimal Inhibitory Concentration (MIC) and Minimal Fungicidal Concentration (MFC). All isolates tested in vitro were sensitive to O. vulgare essential oil. The chromatographic analysis revealed that the main compounds present in the essential oil were 4-terpineol (47.95%), carvacrol (9.42%), thymol (8.42%) and □-terpineol (7.57%). C. albicans isolates obtained from animal mucous membranes exhibited MIC and MFC values of 2.72 μL mL(-1) and 5 μL mL(-1), respectively. MIC and MFC values for C. albicans standard strains were 2.97 μL mL(-1) and 3.54 μL mL(-1), respectively. The MIC and MFC for non-albicans species were 2.10 μL mL(-1) and 2.97 μL mL(-1), respectively. The antifungal activity of O. vulgare essential oil against Candida spp. observed in vitro suggests its administration may represent an alternative treatment for candidiasis.
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http://dx.doi.org/10.1590/S1517-838220100001000018DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3768597PMC
January 2010

Efficacy of terbinafine and itraconazole on a experimental model of systemic sporotrichosis.

Braz J Microbiol 2008 Oct 1;39(4):734-7. Epub 2008 Dec 1.

Programa de Pós-Graduação em Ciências Veterinárias, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul , Porto Alegre, RS , Brasil.

Itraconazole is currently considered the drug of choice to treat the diverse clinical presentation of sporotrichosis. On the other hand terbinafine by virtue of its excellent in vitro activity is under comparative evaluation for its therapeutic potential for a wide range of fungal infections. In this study, our aim was to determine the in vivo efficacy of terbinafine and itraconazole on a experimental model of systemic sporotrichosis. 120 rats Wistar received an injection of 2×10(3) S. schenckii cells by via the lateral tail vein. After 3 days the animals were treated with terbinafine (250mg/kg) and itraconazole (100 mg/kg) and their respective diluents. In our model, terbinafine and itraconazole were effective in reducing the number of clinical lesions and positive organ cultures. There was statistical difference between the groups treated with the antifungals in relation to the control groups (p<0,05) concerning the clinical alterations, anatomic-pathological findings and in the positive organ cultures of the agent, being that the treated animals resulted in the absence and/or reduction of all the evaluated parameters. As for the treatments, terbinafine showed similar or higher activity that itraconazole in the evaluation of the testicle alteration (p=0,0004), as well as in the positive organ cultures of microorganism from the organ (p=0,0142). With these results it is possible to conclude that the antifungals studied are effective in the treatment of experimental systemic sporotrichosis.
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http://dx.doi.org/10.1590/S1517-838220080004000025DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3768454PMC
October 2008

[In vitro susceptibility of isolates of Sporothrix schenckii to terbinafine and itraconazole].

Rev Soc Bras Med Trop 2007 Jan-Feb;40(1):60-2

Programa de Pós-Graduação em Ciências Veterinárias, Universidade Federal do Rio Grande do Sul, Pelotas, RS.

The study objective was to determine the in vitro activity of terbinafine and itraconazole through the microdilution technique in broth (NCCLSM27-A2), adapted for dimorphic fungus, in relation to 12 isolates of Sporothrix schenckii. Six were from feline sporotrichosis, three from human sporotrichosis, one from a dog and two from human isolates originating from Instituto Oswaldo Cruz. The inoculum and antifungal concentrates were distributed on microplates that were incubated at 35 degrees C for five days. Minimum inhibitory concentration readings were made at the end of this period. The MIC for terbinafine ranged from 0.055 microg/ml to 0.109 microg/ml, and the MIC for itraconazole ranged from 0.219 microg/ml to 1.75 microg/ml. For both drugs, the MIC from the isolates from IOC was 0.875 microg/ml. The present study demonstrates the high susceptibility of Sporothrix schenckii to terbinafine. Further studies to correlate the in vitro susceptibility tests with the clinical response of patients with sporotrichosis are needed.
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http://dx.doi.org/10.1590/s0037-86822007000100012DOI Listing
July 2007

Adjuvant effect of green propolis on humoral immune response of bovines immunized with bovine herpesvirus type 5.

Vet Immunol Immunopathol 2007 Mar 14;116(1-2):79-84. Epub 2007 Jan 14.

Centro de Biotecnologia, Universidade Federal de Pelotas (UFPel), Campus Universitário, Caixa Postal 354, 96010-900, Pelotas, RS, Brazil.

Despite recent technological advances in vaccine production, most vaccines depend on the association with adjuvant substances. In this study, propolis, which has been attracting the attention of researchers due to its bioactive properties, was evaluated as an immunological adjuvant. The association of 40mg/dose of an ethanolic extract of green propolis with an inactivated oil vaccine against bovine herpesvirus type 5 (BoHV-5), resulted in a significant increase (P<0.01) in the neutralizing antibody levels, comparing to the bovines that received the same vaccine without propolis. Besides, propolis increased the percentage of animals with high antibody titers (above 32). Phenolic compounds such as artepillin C (3,5-diprenyl-4-hydroxycinnamic acid) and the derivatives of cinnamic acid besides other flavonoid substances were abundant in the propolis extract used, and they could be the main substances with adjuvant action. The effect of the green propolis extract on the humoral immune response can be exploited in the development of new vaccines.
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http://dx.doi.org/10.1016/j.vetimm.2007.01.003DOI Listing
March 2007

Differences in virulence between isolates of feline Sporotrichosis.

Mycopathologia 2005 Aug;160(1):43-9

Programa de Pós-Graduação em Ciências Veterinárias, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Avemda Bento Gonçalves no. 9090-91540-000, Porto Alegre, RS-Brazil.

Sporotrichosis is a chronic subcutaneous mycosis caused by Sporothrix schenckii. This work aimed to evaluate the virulence of two different isolates of S. schenckii from cutaneous (CUT) and systemic (SYS) forms of feline sporotrichosis. A standard inoculum with 2 x 10(3) yeast cells/ml was prepared from each of the isolates. The experimental infection was carried out with 0.1 ml of the inoculum from both isolates and then injected in the paw pads of Swiss albino mice of groups CUT and SYS. The clinical evolution of the disease and the diameter of the lesion at the inoculated sites were evaluated during nine weeks. Four necropsies were done to collect material from the lesions (p < 0.01). Group CUT demonstrated a more evident clinical evolution of the disease from week two to week five; large lesions in the paw pad on week four (p < 0.01); and a higher incidence of lesions in other parts of the body (p < 0.01) than group SYS (p < 0.01). S. schenckii was isolated from the inoculated site in groups SYS and CUT until days 30 and 45, respectively. Granulomas with yeast cells usually localized in the central area were observed in histopathology sections on days 15 and 30 post-inoculations. Those yeast cells decreased on day 45 being absent on day 62 when tissue repair initiated. The results showed that distinct clinical isolates of S. schenckii cause significant differences in the clinical evolution of sporotrichosis.
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http://dx.doi.org/10.1007/s11046-005-6866-xDOI Listing
August 2005