In vitro pharmacokinetics/pharmacodynamics modeling and efficacy against systemic candidiasis in Drosophila melanogaster of a bisaryloxypropanamine derivative.

The number of deaths due to systemic fungal infections is increasing alarmingly, which is aggravated by the limitations of traditional treatments and multidrug resistance. Therefore, the research and development of new therapeutic options against pathogenic fungi is an urgent need. To evaluate the fungicidal activity of a synthetic compound, 1,3-bis-(3,4-dichlorophenoxy)propan-2-aminium chloride (2j), through time-kill studies and pharmacokinetics/pharmacodynamics (PK/PD) modeling. The protective effect of the compound was also evaluated using the Drosophila melanogaster minihost model of candidiasis. Mathematical modeling of time-kill data of compound 2j was performed to obtain PD characteristics. Additionally, Toll-deficient D. melanogaster flies were infected with a Candida albicans strain and treated with 2j. We observed that compound 2j demonstrated a time- and dose-dependent fungicidal effect against Candida spp. and dermatophytes, even at low concentrations, and rapidly achieved kill rates reaching the maximum effect in less than one hour. The efficacy of the compound against systemic candidiasis in D. melanogaster flies was comparable to that achieved by fluconazole. These results support the potential of compound 2j as a systemic antifungal agent candidate and serve as a starting point for further studies involving mammalian animal models.

New insights on evolutionary aspects of Pythium insidiosum and other peronosporaleans.

BACKGROUND: The evolution of pathogenic mechanisms is a major challenge, which requires a thorough comprehension of the phylogenetic relationships of pathogens. Peronosporaleans encompasses a heterogeneous group of oomycetes that includes some animal/human pathogens, like Pythium insidiosum. OBJECTIVE: We analysed here the phylogenetic positioning and other evolutionary aspects related to this species and other peronosporaleans, using a multi-locus approach with one mitochondrial and three nuclear genes. METHODOLOGY: Phylogenetic patterns of 55 oomycetes were inferred by maximum likelihood and Bayesian analysis, and a relaxed molecular clock method was applied to infer the divergence time of some peronosporaleans branches. RESULTS: Pythium insidiosum was monophyletic with a major and polytomous clade of American isolates; however, Pythium spp. was found to be paraphyletic with Phytopythium sp. and Phytophthora spp. In general, peronosporaleans subdivided into four lineages, one of which evidenced a close relationship of P insidiosum, P aphanidermatum and P arrhenomanes. This lineage diverged about 63 million years ago (Mya), whereas P insidiosum diversified at approximately 24 Mya. The divergence of American and Thai isolates seems to have occurred at approximately 17 Mya, with further American diversification at 2.4 Mya. CONCLUSION: Overall, this study clarifies the phylogenetic relationships of P insidiosum regarding other peronosporaleans in a multi-locus perspective, despite previous claims that phylogenomic analyses are needed to accurately infer the patterns and processes related to the evolution of different lineages in this group. Additionally, this is the first time that a molecular clock was applied to study the evolution of P insidiosum.

Oral clioquinol is effective in the treatment of a fly model of Candida systemic infection.

BACKGROUND: Clioquinol was used in the 1950s-1970s as antimicrobial but its oral formulations were withdrawn from the market due to suspected neurotoxicity. Currently, there is possibility of repositioning of oral clioquinol formulations. OBJECTIVES: To evaluate the antifungal activity and toxicological parameters of clioquinol and the other two 8-hydroxyquinoline derivatives using alternative animal models and to study the interaction dynamic of clioquinol with Candida albicans. METHODS: We used Toll-deficient Drosophila melanogaster to test the protective effect of 8-hydroxyquinolines against C. albicans infection. Toxicological parameters were investigated in chicken embryo. A mathematical model-based analysis of the time-kill data of clioquinol was performed to obtain pharmacodynamic characteristics. RESULTS: Clioquinol fully protected D. melanogaster from the infection. The 8-hydroxyquinolines did not cause changes in opening of the beak and movement of the chicken embryo; however, clioquinol and compound 2 increased arterial pulsation. Compound 3 was lethal at 1 mg mL-1 . Effective concentration found in modelling indicated that clioquinol was highly effective against C. albicans (0.306 µg mL-1 ) in easily achievable serum levels; clioquinol rapidly achieved kill rate reaching the maximum effect after 13 hours. CONCLUSIONS: These results support the potential of clioquinol to be used as a systemic antifungal agent.

Genotyping of South American clinical isolates of Pythium insidiosum based on single nucleotide polymorphism-based multiplex PCR / Genotipagem de isolados clínicos de Pythium insidiosum da América do Sul utilizando polimorfismos de nucleotídeo único baseado em PCR multiplex

We aimed to genotype the South American clinical isolates of Pythium insidiosum using the single nucleotide polymorphisms (SNP) of the ribosomal DNA sequences (rDNA). Previously, an SNP-based multiplex-PCR was able to distinguish three different clades of P. insidiosum isolates. Thus, we used this assay to evaluate South American clinical isolates of P. insidiosum (n=32), standard strains from Costa Rica (n=4), Thailand (n=3), Japan (n=1), and India (n=1), a standard strain of Pythium aphanidermatum, and Brazilian environmental isolates of Pythium torulosum, Pythium rhizo-oryzae and Pythium pachycaule voucher (n=3). It was possible to allocate each American P. insidiosum isolate to clade I, the isolates of India, Japan, and Thailand to clade II, and the Thai isolate to clade III. P. aphanidermatum, P.torulosum, P.rhizo-oryzae and P.pachycaule voucher isolates were not amplified. For the first time, a P. insidiosum isolate from Uruguay, South America, was included in molecular analyzes. By SNP-based multiplex-PCR, it was possible to perform the identification and genotyping of the South American isolates of P. insidiosum, demonstrating similar genetic characteristics of these isolates.(AU)

O objetivo deste estudo foi genotipar isolados clínicos de Pythium insidiosum da América do Sul utilizando polimorfismos de nucleotídeo único (SNP) de sequências de rDNA. Anteriormente, um multiplex-PCR baseado em SNP foi capaz de distinguir P. insidiosum em três diferentes clados. Dessa forma, utilizamos este método para avaliar isolados clínicos de P. insidiosum da América do Sul (n=32), cepas padrão da Costa Rica (n=4), Tailândia (n=3), Japão (n=1) e Índia (n=1), uma cepa padrão de Pythium aphanidermatum e isolados ambientais brasileiros de Pythium torulosum; Pythium rhizo-oryzae e Pythium pachycaule voucher (n=3). Os isolados analisados foram alocados aos clados: I (americanos), II (isolados da Índia, Japão e Tailândia), e III (um isolado tailandês). P. aphanidermatum, P.torulosum, P.rhizo-oryzae e P.pachycaule voucher não foram amplificados. Pela primeira vez, um isolado de P. insidiosum do Uruguai foi incluído em análises moleculares. Através da multiplex-PCR baseada em SNP, foi possível realizar a identificação e genotipagem dos isolados sul-americanos de P. insidiosum, demonstrando características genéticas semelhantes entre esses isolados.(AU)

In vitro susceptibility of chromoblastomycosis agents to antifungal drugs: A systematic review.

Chromoblastomycosis (CBM) is a chronic granulomatous mycosis caused by dematiaceous fungi that affects cutaneous and subcutaneous tissues. The standard antifungal drug for treatment is itraconazole, followed by terbinafine. However, cure rates vary from 15% to 80% when these drugs are used as monotherapy. A systematic review of the in vitro susceptibility of CBM agents to antifungal drugs, alone and in combination, was conducted using the Cochrane methodology. Forty-seven search terms were included in the PICOS method of searching electronic databases. The search resulted in 35 studies, of which 8 evaluated antifungal drugs in combination. Based on minimum inhibitory concentrations (MICs), posaconazole, terbinafine, itraconazole and voriconazole were, in descending order, the most effective antifungal drugs against CBM agents in vitro. In drug combination studies, only terbinafine-voriconazole and itraconazole-caspofungin showed 100% synergy for Fonsecaea pedrosoi, Exophiala jeanselmei and Phialophora verrucosa. However, none of the combinations studied showed antagonism.

Melanin: Quantification and protection against oxidative stress in chromoblastomycosis agents.

Chromoblastomycosis (CBM) is a chronic cutaneous and subcutaneous infection caused by melanized fungal species. We quantified the extractable melanin of 77 strains of CBM agents distributed within five genera. Moreover, resistance to oxidative stress was evaluated in strains exposed or not to the melanin inhibitor tricyclazole. The median percentage of melanin mass extracted from dry fungal mass varied from 0.69 (Rhinocladiella similis) to 3.81 (Phialophora americana). Inhibition of melanin synthesis decreased survival rates to hydrogen peroxide. Together, these data highlight the importance of melanin in CBM agents.

Genotyping of South American clinical isolates of Pythium insidiosum based on single nucleotide polymorphism-based multiplex PCR / Genotipagem de isolados clínicos de Pythium insidiosum da América do Sul utilizando polimorfismos de nucleotídeo único baseado em PCR multiplex

ABSTRACT: We aimed to genotype the South American clinical isolates of Pythium insidiosum using the single nucleotide polymorphisms (SNP) of the ribosomal DNA sequences (rDNA). Previously, an SNP-based multiplex-PCR was able to distinguish three different clades of P. insidiosum isolates. Thus, we used this assay to evaluate South American clinical isolates of P. insidiosum (n=32), standard strains from Costa Rica (n=4), Thailand (n=3), Japan (n=1), and India (n=1), a standard strain of Pythium aphanidermatum, and Brazilian environmental isolates of Pythium torulosum, Pythium rhizo-oryzae and Pythium pachycaule voucher (n=3). It was possible to allocate each American P. insidiosum isolate to clade I, the isolates of India, Japan, and Thailand to clade II, and the Thai isolate to clade III. P. aphanidermatum, P.torulosum, P.rhizo-oryzae and P.pachycaule voucher isolates were not amplified. For the first time, a P. insidiosum isolate from Uruguay, South America, was included in molecular analyzes. By SNP-based multiplex-PCR, it was possible to perform the identification and genotyping of the South American isolates of P. insidiosum, demonstrating similar genetic characteristics of these isolates.

RESUMO: O objetivo deste estudo foi genotipar isolados clínicos de Pythium insidiosum da América do Sul utilizando polimorfismos de nucleotídeo único (SNP) de sequências de rDNA. Anteriormente, um multiplex-PCR baseado em SNP foi capaz de distinguir P. insidiosum em três diferentes clados. Dessa forma, utilizamos este método para avaliar isolados clínicos de P. insidiosum da América do Sul (n=32), cepas padrão da Costa Rica (n=4), Tailândia (n=3), Japão (n=1) e Índia (n=1), uma cepa padrão de Pythium aphanidermatum e isolados ambientais brasileiros de Pythium torulosum; Pythium rhizo-oryzae e Pythium pachycaule voucher (n=3). Os isolados analisados foram alocados aos clados: I (americanos), II (isolados da Índia, Japão e Tailândia), e III (um isolado tailandês). P. aphanidermatum, P.torulosum, P.rhizo-oryzae e P.pachycaule voucher não foram amplificados. Pela primeira vez, um isolado de P. insidiosum do Uruguai foi incluído em análises moleculares. Através da multiplex-PCR baseada em SNP, foi possível realizar a identificação e genotipagem dos isolados sul-americanos de P. insidiosum, demonstrando características genéticas semelhantes entre esses isolados.

In vitro interactions of azoles and echinocandins against clinical strains of Aspergillus flavus.

Combinations of an azole (itraconazole, voriconazole, or posaconazole) with an echinocandin (caspofungin, micafungin, or anidulafungin) were tested against 20 clinical isolates of Aspergillus flavus according to EUCAST guidelines. The interactions were determined using two endpoints-minimal effective concentration (MEC) and minimal inhibitory concentration (MIC)-via calculation of the fractional inhibitory concentration (FIC) index. A higher prevalence of synergistic interactions was observed for MIC, whereas indifference was the most frequent outcome according to MEC among the 20 strains. Combined treatment of A. flavus with these two important classes of antifungals should be explored further in in vivo studies.

In vitro antifungal susceptibility of clinical and environmental isolates of Aspergillus fumigatus and Aspergillus flavus in Brazil

ABSTRACT The in vitro susceptibility of 105 clinical and environmental strains of Aspergillus fumigatus and Aspergillus flavus to antifungal drugs, such as amphotericin B, azoles, and echinocandins was evaluated by the broth microdilution method proposed by the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Following the EUCAST-proposed breakpoints, 20% and 25% of the clinical and environmental isolates of A. fumigatus, respectively, were found to be resistant to itraconazole (Minimal Inhibitory Concentration, MIC > 2.0 mg/L). Voriconazole showed good activity against A. fumigatus and A. flavus strains, except for one clinical strain of A. fumigatus whose MIC was 4.0 mg/L. Posaconazole (≤0.25 mg/L) also showed appreciable activity against both species of Aspergillus, except for six A. fumigatus strains with relatively higher MICs (0.5 mg/L). The MICs for Amphotericin B ranged from 0.06 to 1.0 mg/L for A. fumigatus, but were much higher (0.5-8.0 mg/L) for A. flavus. Among the echinocandins, caspofungin showed a geometric mean of 0.078 and 0.113 against the clinical and environmental strains of A. flavus, respectively, but had elevated minimal effective concentrations (MECs) for seven of the A. fumigatus strains. Anidulafungin and micafungin exhibited considerable activity against both A. fumigatus and A. flavus isolates, except for one environmental isolate of A. fumigatus that showed an MEC of 1 mg/L to micafungin. Our study proposes that a detailed investigation of the antifungal susceptibility of the genus Aspergillus from different regions of Brazil is necessary for establishing a response profile against the different classes of antifungal agents used in the treatment of aspergillosis.

In vitro antifungal susceptibility of clinical and environmental isolates of Aspergillus fumigatus and Aspergillus flavus in Brazil.

The in vitro susceptibility of 105 clinical and environmental strains of Aspergillus fumigatus and Aspergillus flavus to antifungal drugs, such as amphotericin B, azoles, and echinocandins was evaluated by the broth microdilution method proposed by the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Following the EUCAST-proposed breakpoints, 20% and 25% of the clinical and environmental isolates of A. fumigatus, respectively, were found to be resistant to itraconazole (Minimal Inhibitory Concentration, MIC>2.0mg/L). Voriconazole showed good activity against A. fumigatus and A. flavus strains, except for one clinical strain of A. fumigatus whose MIC was 4.0mg/L. Posaconazole (≤0.25mg/L) also showed appreciable activity against both species of Aspergillus, except for six A. fumigatus strains with relatively higher MICs (0.5mg/L). The MICs for Amphotericin B ranged from 0.06 to 1.0mg/L for A. fumigatus, but were much higher (0.5-8.0mg/L) for A. flavus. Among the echinocandins, caspofungin showed a geometric mean of 0.078 and 0.113 against the clinical and environmental strains of A. flavus, respectively, but had elevated minimal effective concentrations (MECs) for seven of the A. fumigatus strains. Anidulafungin and micafungin exhibited considerable activity against both A. fumigatus and A. flavus isolates, except for one environmental isolate of A. fumigatus that showed an MEC of 1mg/L to micafungin. Our study proposes that a detailed investigation of the antifungal susceptibility of the genus Aspergillus from different regions of Brazil is necessary for establishing a response profile against the different classes of antifungal agents used in the treatment of aspergillosis.

Gamma-Decanolactone Improves Biochemical Parameters Associated with Pilocarpine-Induced Seizures in Male Mice.

BACKGROUND AND OBJECTIVE: Gamma-decanolactone (GD) is a monoterpene effective against seizures induced by pentylenetetrazole. The mechanism of action of GD is likely to be via glutamate antagonism. GD also inhibits intracellular reactive oxygen species (ROS) generation and the lipopolysaccharide-induced expression of inducible nitric oxide synthase (iNOS) and tumor necrosis factor-alpha (TNF-α) in vitro. Considering the neuropharmacological profile of GD studied so far, we investigated the effect of intraperitoneal administration of GD 100 and 300 mg/kg on pilocarpine (PIL)-induced status epilepticus (SE) in mice. METHODS: GD was administered 30 min before PIL. Behavioral (latency to first seizure and the percentage of clonic forelimb seizures), biochemical, and oxidative stress parameters were evaluated. DNA damage in the cerebral cortex of mice was assessed using the comet assay and mutagenic activity of GD was evaluated using Salmonella/microsome assay in TA100, TA98, TA97a, TA102, and TA1535 strains, with and without metabolic activation (S9 mix). RESULTS: The behavioral results showed that only the latency to the first clonic seizure increased in the groups treated with GD 300 mg/kg, but not when the animals received GD 100 mg/kg. Both GD doses were able to increase superoxide dismutase and catalase activities, inducing a decrease in ROS and nitrite production and in DNA damage in the cerebral cortex. GD was not able to induce base pair substitution and frameshift mutations in the absence or in the presence of metabolic activation. CONCLUSION: These findings demonstrate that GD does not improve behavioral parameters in the PIL model, but it was able to protect seizure-related oxidative stress and DNA damage in mice, without inducing gene mutations.

In vitro effects of Blepharocalyx salicifolius (H.B.K.) O. Berg on the viability of Echinococcus ortleppi protoscoleces.

Scolicidal agents are important in the treatment of cystic echinococcosis. This study evaluated the scolicidal activity of the plant Blepharocalyx salicifolius (H.B.K.) Berg against Echinococcus ortleppi protoscoleces. The parasite species was identified by amplifying a fragment of the gene cytochrome c oxidase subunit 1 (COX 1). B. salicifolius crude extract at concentrations of 100, 200, 300 and 400 mg/mL was analyzed at different times (5, 10, 15, 30, 45 and 60 min). N-butanol and ethyl acetate fractions (100 and 200 mg/ mL) were also analyzed at 5, 10, 15 and 30 min. Both fractions showed 100% scolicidal activity at the concentration of 200 mg/mL at 5 min. Gallic acid, identified as the major compound of the ethyl acetate fraction- was responsible for the observed scolicidal activity. The results showed that crude extract and fractions of B. salicifolius have scolicidal effect against E. ortleppi protoscoleces.

Antifungal activity of synthetic antiseptics and natural compounds against Candida dubliniensis before and after in vitro fluconazole exposure.

INTRODUCTION:: This study evaluated the susceptibilities of oral candidiasis-derived Candida albicans, fluconazole-resistant (FR) Candida dubliniensis, and fluconazole-susceptible (FS) C. dubliniensis to synthetic antiseptics [chlorhexidine gluconate (CHX), cetylpyridinium chloride (CPC), and triclosan (TRC)] and natural compounds (carvacrol, eugenol and thymol). METHODS:: Susceptibility tests were performed based on the M27-A3 reference method. The fluconazole-resistant C. dubliniensis strains were obtained after prolonged in vitro exposure to increasing fluconazole concentrations. The geometric mean values for minimum inhibitory concentrations and minimum fungicidal concentrations were compared among the groups. RESULTS:: Fluconazole-susceptible C. dubliniensis was more sensitive to CPC and TRC than FR C. dubliniensis and C. albicans were. However, eugenol and thymol were more active against FR C. dubliniensis. The fungicidal activities of CHX and TRC were similar for the three groups, and FR C. dubliniensis and C. albicans had similar sensitivities to CPC. CONCLUSIONS:: The resistance of C. dubliniensis to fluconazole affects its sensitivity the synthetic antiseptics and natural compounds that were tested.

Antifungal activity of synthetic antiseptics and natural compounds against Candida dubliniensis before and after in vitro fluconazole exposure

ABSTRACT INTRODUCTION: This study evaluated the susceptibilities of oral candidiasis-derived Candida albicans, fluconazole-resistant (FR) Candida dubliniensis, and fluconazole-susceptible (FS) C. dubliniensis to synthetic antiseptics [chlorhexidine gluconate (CHX), cetylpyridinium chloride (CPC), and triclosan (TRC)] and natural compounds (carvacrol, eugenol and thymol). METHODS: Susceptibility tests were performed based on the M27-A3 reference method. The fluconazole-resistant C. dubliniensis strains were obtained after prolonged in vitro exposure to increasing fluconazole concentrations. The geometric mean values for minimum inhibitory concentrations and minimum fungicidal concentrations were compared among the groups. RESULTS: Fluconazole-susceptible C. dubliniensis was more sensitive to CPC and TRC than FR C. dubliniensis and C. albicans were. However, eugenol and thymol were more active against FR C. dubliniensis. The fungicidal activities of CHX and TRC were similar for the three groups, and FR C. dubliniensis and C. albicans had similar sensitivities to CPC. CONCLUSIONS: The resistance of C. dubliniensis to fluconazole affects its sensitivity the synthetic antiseptics and natural compounds that were tested.

Pre- and postnatal evaluation of offspring rats exposed to Origanum vulgare essential oil during mating, gestation and lactation / Avaliação pré e pós-natal da prole de ratos expostos ao óleo essencial de Origanum vulgare durante acasalamento, gestação e lactação

Despite the increasing use of oregano (Origanum vulgare L.) essential oil for therapeutic purposes, pre- and postnatal development of animals offspring exposed to this oil has not yet been evaluated. In line with previous concerns of genotoxicity, in this study adult rats were exposed to different doses of oregano essential oil (3, 9 and 27% vol/vol) during pre-mating, mating, gestation and lactation. Prenatal screening included fetal development and uterine inspection, where the reproductive rate of females such as breeding, pregnancy, delivery, viability and post-implantation loss rate were measured. Postnatal evaluation of rat offspring included motor development, neuroendocrine and behavioral assessment. Body weight of rat dams and signs of dystocia were evaluated daily. Development of physic characteristics and reflex tests of puppies were also assessed. Additionally, these rats, when adults, were submitted to sexual and open field behavioral tests. The main differences among the groups were observed in the indices of mating, pregnancy and post-implantation loss (P < 0.01). Results demonstrated that the treatment of parental generation with oregano essential oil has the potential to affect the developing fetuses at the highest dose used, but without causing maternal toxicity and changes in general behavior and development of the progeny.

Apesar do aumento do uso do óleo essencial de orégano (Origanum vulgare L.) para fins terapêuticos, o desenvolvimento pré e pós-natal da progênie de animais expostos a este óleo ainda não foi avaliado. Partindo das suspeitas prévias de genotoxicidade desse óleo, neste estudo, ratos Wistar adultos foram expostos a diferentes doses do óleo essencial de orégano (3, 9 e 27% vol/vol) durante o acasalamento, a gestação e a lactação. Para a avaliação pré-natal, o desenvolvimento gestacional foi observado e os úteros inspecionados, assim como os índices reprodutivos das fêmeas, como a taxa de acasalamento, de gestação, parto e perdas pós-implantação. Na avaliação pós-natal, observou-se o desenvolvimento motor, neuroendócrino e comportamental da prole. Observou-se, diariamente, o peso das ratas e sinais de distocia. Após o parto, as características de desenvolvimento e testes de reflexos dos filhotes foram avaliadas, enquanto que, na puberdade, foram realizadas análises histopatológicas e dosagem hormonal. Adicionalmente, na idade adulta, esses ratos foram submetidos ao teste de comportamento em campo aberto e ao comportamento sexual. As principais diferenças entre os grupos foram nos índices de acasalamento, de gestação e de perdas pós-implantação (P < 0,01). Os resultados demonstram que o tratamento da geração parental com óleo essencial de orégano tem potencial para afetar os índices reprodutivos das ratas e o desenvolvimento dos fetos na maior dose utilizada, mas sem causar toxicidade materna e alterações no desenvolvimento geral e comportamental da sua progênie.

Pre- and postnatal evaluation of offspring rats exposed to Origanum vulgare essential oil during mating, gestation and lactation / Avaliação pré e pós-natal da prole de ratos expostos ao óleo essencial de Origanum vulgare durante acasalamento, gestação e lactação

Despite the increasing use of oregano (Origanum vulgare L.) essential oil for therapeutic purposes, pre- and postnatal development of animals offspring exposed to this oil has not yet been evaluated. In line with previous concerns of genotoxicity, in this study adult rats were exposed to different doses of oregano essential oil (3, 9 and 27% vol/vol) during pre-mating, mating, gestation and lactation. Prenatal screening included fetal development and uterine inspection, where the reproductive rate of females such as breeding, pregnancy, delivery, viability and post-implantation loss rate were measured. Postnatal evaluation of rat offspring included motor development, neuroendocrine and behavioral assessment. Body weight of rat dams and signs of dystocia were evaluated daily. Development of physic characteristics and reflex tests of puppies were also assessed. Additionally, these rats, when adults, were submitted to sexual and open field behavioral tests. The main differences among the groups were observed in the indices of mating, pregnancy and post-implantation loss (P < 0.01). Results demonstrated that the treatment of parental generation with oregano essential oil has the potential to affect the developing fetuses at the highest dose used, but without causing maternal toxicity and changes in general behavior and development of the progeny.(AU)

Apesar do aumento do uso do óleo essencial de orégano (Origanum vulgare L.) para fins terapêuticos, o desenvolvimento pré e pós-natal da progênie de animais expostos a este óleo ainda não foi avaliado. Partindo das suspeitas prévias de genotoxicidade desse óleo, neste estudo, ratos Wistar adultos foram expostos a diferentes doses do óleo essencial de orégano (3, 9 e 27% vol/vol) durante o acasalamento, a gestação e a lactação. Para a avaliação pré-natal, o desenvolvimento gestacional foi observado e os úteros inspecionados, assim como os índices reprodutivos das fêmeas, como a taxa de acasalamento, de gestação, parto e perdas pós-implantação. Na avaliação pós-natal, observou-se o desenvolvimento motor, neuroendócrino e comportamental da prole. Observou-se, diariamente, o peso das ratas e sinais de distocia. Após o parto, as características de desenvolvimento e testes de reflexos dos filhotes foram avaliadas, enquanto que, na puberdade, foram realizadas análises histopatológicas e dosagem hormonal. Adicionalmente, na idade adulta, esses ratos foram submetidos ao teste de comportamento em campo aberto e ao comportamento sexual. As principais diferenças entre os grupos foram nos índices de acasalamento, de gestação e de perdas pós-implantação (P < 0,01). Os resultados demonstram que o tratamento da geração parental com óleo essencial de orégano tem potencial para afetar os índices reprodutivos das ratas e o desenvolvimento dos fetos na maior dose utilizada, mas sem causar toxicidade materna e alterações no desenvolvimento geral e comportamental da sua progênie.(AU)

In vitro and ex vivo activity of Melaleuca alternifolia against protoscoleces of Echinococcus ortleppi.

Cystic echinococcosis is a zoonotic disease of difficult diagnosis and treatment. The use of protoscolicidal agents in procedures is of utmost importance for treatment success. This study was aimed at analysing the in vitro and ex vivo activity of Melaleuca alternifolia oil (tea tree oil - TTO), its nanoemulsion formulation (NE-TTO) and its major component (terpinen-4-ol) against Echinococcus ortleppi protoscoleces obtained from cattle. Concentrations of 2·5, 5 and 10 mg mL-1 of TTO, 10 mg mL-1 of NE-TTO and 1, 1·5 and 2 mg mL-1 of terpinen-4-ol were evaluated in vitro against protoscoleces at 5, 10, 15 and 30 min. TTO was also injected directly into hydatid cysts (ex vivo analysis, n = 20) and the viability of protoscoleces was evaluated at 5, 15 and 30 min. The results indicated protoscolicidal effect at all tested formulations and concentrations. Terpinen-4-ol (2 mg mL-1) activity was superior when compared with the highest concentration of TTO. NE-TTO reached a gradual protoscolicidal effect. TTO at 20 mg mL-1 showed 90% protoscolicidal action in hydatid cysts at 5 min. The results showed that TTO affects the viability of E. ortleppi protoscoleces, suggesting a new protoscolicidal option to the treatment of cystic equinococcosis.

Seroprevalence of Pythium insidiosum infection in equine in Rio Grande do Sul, Brazil / Soroprevalência da infecção por Pythium insidiosum em equinos no estado do Rio Grande do Sul

ABSTRACT: An epidemiological survey was carried out by performing an Enzyme Linked Immuno Sorbent Assay (ELISA) test to determine the seroprevalence of Pythium insidiosum infection in equine in Rio Grande do Sul State (RS), Brazil. The serological study covered seven geographical regions of RS, classified according to the Instituto Brasileiro de Geografia e Estatística (IBGE). The samples were obtained from official veterinary service (Serviço Veterinário Oficial, SVO) linked to the Secretaria da Agricultura, Pecuária e Agronegócio of RS (SEAPA-RS) to proceed the investigation of equine infectious anemia in 2014. Samples were collected during the months of September and October of 2013, covering the seven geographical regions of RS, and totalized 1,002 serum samples. The seroprevalence for P. insidiosum in RS was 11.1% (CI95% 9.23 to 13.22). The relative risk (RR) of the presence of antibodies anti-P. insidiosum was in the regions Southeast 11.17 (CI95%, 4.65 to 26.8), Porto Alegre 4.62 (CI95%, 1.70 to 12.55), Southwest 11.17 (CI95%, 4.65 to 26.8) and Northwestern 3.72 (CI95%, 1.52 to 9.09). The highest prevalence (69.1%) was observed in females with RR of 1.59 (CI95%, 1.11 to 2.27). When the presence of dams was evaluated, the seropositivity was evident in 74.4%, presenting an association of 2.13 (CI95%, 1.16 to 3.91) compared to farms without dams. In properties with veterinary assistance, the frequency of 72.7% and RR of 3.04 (CI95%,, 1,85 to 4,98) of seropositivity were observed. Due to the importance of pythiosis in horse herds, this study highlights the presence of anti-P. insidiosum antibodies in horses in RS, Brazil.

RESUMO: Um levantamento soroepidemiológico foi realizado através do teste de ELISA indireto para determinar a soroprevalência da infecção por Pythium insidiosum em equinos no estado do Rio Grande do Sul (RS), classificadas de acordo com o Instituto Brasileiro de Geografia e Estatística (IBGE). As amostras utilizadas eram provenientes do cadastro das propriedades do Serviço Veterinário Oficial (SVO), da Secretaria da Agricultura, Pecuária e Agronegócio do RS (SEAPA-RS), coletadas para o inquérito da anemia infecciosa equina de 2014. As coletas foram realizadas durante os meses de setembro e outubro de 2013, abrangendo as sete mesorregiões geográficas do RS, e totalizaram 1.002 amostras de soro. Do total das amostras testadas, 11.1% (CI95% 9.23 to 13.22) foram soropositivas para P. insidiosum. Constatou-se o risco relativo (RR) da presença de anticorpos anti-P. insidiosum nas regiões Sudeste 11,17(IC95%, 4,65-26,8), Porto Alegre 4,62 (IC95%, 1,70-12,55), Sudoeste 11,17 (IC95%, 4,65-26,8) e Noroeste 3,72 (IC95%, 1,52-9,09). Observou-se a maior soroprevalência (69,1%) em fêmeas com RR de 1,59 (IC95%, 1,11-2,27). Quanto à presença de açudes, evidenciou-se soropositividade em 74,4% das propriedades, apresentando associação de 2,13 (IC95%,1,16-3,91) em comparação com propriedades sem açude. Em propriedades com assistência veterinária, foi verificada a frequência de 72,7% e RR de 3,04 (IC95%,1,85-4,98). Diante da relevância da pitiose em rebanhos equinos, destaca-se a presença de anticorpos anti-P. insidiosum em equinos no estado do RS.

Seroprevalence of Pythium insidiosum infection in equine in Rio Grande do Sul, Brazil / Soroprevalência da infecção por Pythium insidiosum em equinos no estado do Rio Grande do Sul

An epidemiological survey was carried out by performing an Enzyme Linked Immuno Sorbent Assay (ELISA) test to determine the seroprevalence of Pythium insidiosum infection in equine in Rio Grande do Sul State (RS), Brazil. The serological study covered seven geographical regions of RS, classified according to the Instituto Brasileiro de Geografia e Estatística (IBGE). The samples were obtained from official veterinary service (Serviço Veterinário Oficial, SVO) linked to the Secretaria da Agricultura, Pecuária e Agronegócio of RS (SEAPA-RS) to proceed the investigation of equine infectious anemia in 2014. Samples were collected during the months of September and October of 2013, covering the seven geographical regions of RS, and totalized 1,002 serum samples. The seroprevalence for P. insidiosum in RS was 11.1% (CI95% 9.23 to 13.22). The relative risk (RR) of the presence of antibodies anti-P. insidiosum was in the regions Southeast 11.17 (CI95%, 4.65 to 26.8), Porto Alegre 4.62 (CI95%, 1.70 to 12.55), Southwest 11.17 (CI95%, 4.65 to 26.8) and Northwestern 3.72 (CI95%, 1.52 to 9.09). The highest prevalence (69.1%) was observed in females with RR of 1.59 (CI95%, 1.11 to 2.27). When the presence of dams was evaluated, the seropositivity was evident in 74.4%, presenting an association of 2.13 (CI95%, 1.16 to 3.91) compared to farms without dams. In properties with veterinary assista(AU)

Um levantamento soroepidemiológico foi realizado através do teste de ELISA indireto para determinar a soroprevalência da infecção por Pythium insidiosum em equinos no estado do Rio Grande do Sul (RS), classificadas de acordo com o Instituto Brasileiro de Geografia e Estatística (IBGE). As amostras utilizadas eram provenientes do cadastro das propriedades do Serviço Veterinário Oficial (SVO), da Secretaria da Agricultura, Pecuária e Agronegócio do RS (SEAPA-RS), coletadas para o inquérito da anemia infecciosa equina de 2014. As coletas foram realizadas durante os meses de setembro e outubro de 2013, abrangendo as sete mesorregiões geográficas do RS, e totalizaram 1.002 amostras de soro. Do total das amostras testadas, 11.1% (CI95% 9.23 to 13.22) foram soropositivas para P. insidiosum. Constatou-se o risco relativo (RR) da presença de anticorpos anti-P. insidiosum nas regiões Sudeste 11,17(IC95%, 4,65-26,8), Porto Alegre 4,62 (IC95%, 1,70-12,55), Sudoeste 11,17 (IC95%, 4,65-26,8) e Noroeste 3,72 (IC95%, 1,52-9,09). Observou-se a maior soroprevalência (69,1%) em fêmeas com RR de 1,59 (IC95%, 1,11-2,27). Quanto à presença de açudes, evidenciou-se soropositividade em 74,4% das propriedades, apresentando associação de 2,13 (IC95%,1,16-3,91) em comparação com propriedades sem açude. Em propriedades com assistência veterinária, foi verificada a frequência de 72,7% e RR de 3,04 (IC95%,1(AU)

Syzygium cumini seed extract ameliorates adenosine deaminase activity and biochemical parameters but does not alter insulin sensitivity and pancreas architecture in a short-term model of diabetes.

BACKGROUND: The effects of the aqueous seed extract of Syzygium cumini (ASc) in a short-term model of diabetes in rats are little explored. The present study was designed to evaluate the effect of the ASc on adenosine deaminase (ADA) activity and on biochemical and histopathological parameters in diabetic rats. METHODS: ASc (100 mg/kg) was administered for 21 days in control and streptozotocin (STZ)-induced (60 mg/kg) diabetic rats. ADA activity, lipoperoxidation (cerebral cortex, kidney, liver and pancreas) and biochemical (serum) and histopathological (pancreas) parameters were evaluated. RESULTS: The main findings in this short-term model of Diabetes mellitus (DM) were that the ASc (i) significantly reverted the increase of ADA activity in serum and kidney; (ii) ameliorated the lipoperoxidation in the cerebral cortex and pancreas of the diabetic group; (iii) demonstrated hypolipidemic and hypoglycemic properties and recovered the liver glycogen; and iv) prevented the HOMA-IR index increase in the diabetic group. Therefore, the ASc can be a positive factor for increasing the availability of substrates with significant protective actions, such as adenosine. Moreover, by maintaining glycogen and HOMA-IR levels, the extract could modulate the hyperglycemic state through the direct peripheral glucose uptake. CONCLUSIONS: Our data revealed that the short-term treatment with ASc has an important protective role under pathophysiological conditions caused by the early stage of DM. These results enhance our understanding of the effect of the ASc on the purinergic system in DM.