A survey of tick-borne pathogens in dogs and their ticks in the Pantanal biome, Brazil.

Tick and blood samples collected from domestic dogs in the Brazilian Pantanal were tested by molecular methods for the presence of tick-borne protozoa and bacteria. Among 320 sampled dogs, 3.13% were infected by Babesia vogeli (Piroplasmida: Babesiidae), 8.75% by Hepatozoon canis (Eucoccidiorida: Hepatozoidae), 7.19% by Anaplasma platys (Rickettsiales: Anaplasmataceae), and 0.94% by an unclassified Anaplasma sp. In three tick species collected from dogs, the following tick-borne agents were detected: (a) B. vogeli, An. platys and Ehrlichia canis (Rickettsiales: Anaplasmataceae), infecting Rhipicephalus sanguineus sensu lato (Ixodida: Ixodidae) ticks; (b) H. canis, an unclassified Anaplasma sp. and Rickettsia amblyommii (Rickettsiales: Rickettsiaceae), infecting Amblyomma cajennense sensu lato (Ixodida: Ixodidae) ticks, and (c) Rickettsia sp. strain Atlantic rainforest, an emerging human pathogen, infecting Amblyomma ovale ticks. Molecular analysis, based on a mitochondrial gene, revealed that the Am. cajennense s.l. ticks of the present study corresponded to Amblyomma sculptum, a member of the Am. cajennense species complex, and that Rh. sanguineus s.l. belonged to the tropical lineage. Whereas dogs are exposed to a number of tick-borne bacterial and protozoan agents in the Pantanal biome, humans are potentially exposed to infection by spotted fever group rickettsiae (e.g. R. amblyommii and Rickettsia sp. strain Atlantic rainforest) because both Am. sculptum and Am. ovale are among the most important human-biting ticks in Brazil.

Can the same species of Platynosomum (Trematoda: Dicrocoeliidae) infect both mammalian and avian hosts?

The importance of platynosomiasis has increased in feline veterinary practice, but aspects related to the specificity of Platynosomum spp. in definitive hosts requires further study. Although morphological traits suggest that the same species, P. illiciens, may infect both birds and mammals, the synonymies previously proposed have not been widely accepted, likely because host specificity is assumed. In addition, the name P. fastosum has frequently been used for parasites recovered from mammals. In the present study, metacercariae (n= 100/animal) of P. illiciens recovered from lizards (Hemidactylus mabouia) in Brazil were fed to Australian parakeets (Melopsittacus undulatus) and mice. Two parasites were recovered from the liver of one M. undulatus specimen during a necropsy that was performed 105 days after infection, and all mice were found to be infected with 37 ± 12 (18-48) parasites. The morphology of the P. illiciens obtained from the parakeet was similar to that of parasites obtained from mice and those described previously from naturally infected birds and mammals. Non-specificity of P. illiciens in hosts is discussed briefly, based on the parasitological and morphological results obtained during the avian experimental platynosomiasis and the epidemiology and geographical distribution of this parasite.

Phenotypic plasticity of male Schistosoma mansoni from the peritoneal cavity and hepatic portal system of laboratory mice and hamsters.

Morphometric analysis of Schistosoma mansoni male worms obtained from AKR/J and Swiss mice was carried out. Rodents infected by the intraperitoneal route with 80 cercariae of the schistosome (LE strain) were killed by cervical dislocation at 45 and 60 days post-infection and both peritoneal lavage and perfusion of the portal system were performed for the recovery of adult worms. Characteristics including total body length, the distance between oral and ventral suckers, extension of testicular mass and the number of testes were considered in the morphological analysis. Changes that occurred in S. mansoni recovered from the peritoneal cavity or from the portal system of AKR/J and Swiss mice included total body length and reproductive characteristics. Significant morphometric alterations were also observed when worms recovered from the portal system of both strains of mice were compared with the schistosomes obtained from hamsters (Mesocricetus auratus), the vertebrate host in which the LE strain had been adapted and maintained by successive passages for more than four decades. The present results reinforce the idea that S. mansoni has high plastic potential and adaptive capacity.

Random regression models using Legendre orthogonal polynomials to evaluate the milk production of Alpine goats.

The objective of this study was to identify the best random regression model using Legendre orthogonal polynomials to evaluate Alpine goats genetically and to estimate the parameters for test day milk yield. On the test day, we analyzed 20,710 records of milk yield of 667 goats from the Goat Sector of the Universidade Federal de Viçosa. The evaluated models had combinations of distinct fitting orders for polynomials (2-5), random genetic (1-7), and permanent environmental (1-7) fixed curves and a number of classes for residual variance (2, 4, 5, and 6). WOMBAT software was used for all genetic analyses. A random regression model using the best Legendre orthogonal polynomial for genetic evaluation of milk yield on the test day of Alpine goats considered a fixed curve of order 4, curve of genetic additive effects of order 2, curve of permanent environmental effects of order 7, and a minimum of 5 classes of residual variance because it was the most economical model among those that were equivalent to the complete model by the likelihood ratio test. Phenotypic variance and heritability were higher at the end of the lactation period, indicating that the length of lactation has more genetic components in relation to the production peak and persistence. It is very important that the evaluation utilizes the best combination of fixed, genetic additive and permanent environmental regressions, and number of classes of heterogeneous residual variance for genetic evaluation using random regression models, thereby enhancing the precision and accuracy of the estimates of parameters and prediction of genetic values.

Current applications of oogram methodology in experimental schistosomiasis; fecundity of female Schistosoma mansoni and egg release in the intestine of AKR/J mice following immunomodulatory treatment with pentoxifylline.

Oogram methodology permitted a direct assessment of the fecundity of Schistosoma mansoni and the passage of parasite eggs from mesenteric vessels into the intestinal lumen in AKR/J mice that had been infected via the intraperitoneal route with 60 cercariae of the trematode and later subjected to short-term subcutaneous treatments with pentoxifylline (PTX). The administration of PTX did not change oviposition kinetics, the individual fecundity of female parasites (as determined by the study of the ratio of second-stage S. mansoni eggs per g intestine/number of female parasites recovered from the portal system), nor the number of mature and dead eggs retained in the host tissue, though the drug has known immunomodulatory effects, as shown previously in experimental schistosomiasis. A better appraisal was also carried out, including the study of statistical parameters, concerning the utilization of the ratios of each stage of immature eggs (first to fourth) per g intestine/number of female worms from the portal system. The second-stage eggs had the lowest variability, confirming that the utilization of this stage as an indicator of the individual fecundity of parasite females is indeed viable. In the light of our findings, current uses of oogram methodology are discussed. Moreover, additional consideration is given to data obtained in the present study concerning intraperitoneal infection with S. mansoni cercariae in both untreated and treated mice of the AKR/J strain, such as the recovery of mature worms, eggs and free granulomas from the peritoneal cavity of these rodents.

Apoptotic mechanisms are involved in the death of Strongyloides venezuelensis after triggering of nitric oxide.

Despite progress in understanding the role of nitric oxide (NO) in the pathogenesis of helminth infections, the role in strongyloidosis is unknown. Firstly, we studied the production of NO in mice infected with Strongyloides venezuelensis as well as in macrophage cultures stimulated with parasite antigens. Somatic larvae 3 (L3) and excretory-secretory female antigens stimulate specific NO production measured by Griess reaction and expression of inducible NO synthase by RT-PCR and quantitative PCR. Moreover, mice infected with S. venezuelensis produce NO in migration stages. Secondly, we analysed the effect of NO production on L3 and females of S. venezuelensis using NO donors such as diethylenetriamine and 3,3-bis(aminoethyl)-1-hydroxy-2-oxo-1-triazene. Parasites died after NO donor treatment in a dose-dependent manner. Finally, apoptotic mechanisms are involved in the death of S. venezuelensis larvae.

Genetic and environmental factors that influence production and quality of milk of Alpine and Saanen goats.

Data from 1279 lactations of 783 Alpine and Saanen goats of the herd of our university in Minas Gerais, Brazil, were used to study environmental effects on and to estimate genetic parameters for milk production until 270 days of lactation (MP270) and for production and percentages of fat (PFAT and %FAT), protein (PPROT and %PROT), lactose (PLACT and %LACT), and total dry extract (PEXTR and %EXTR). Environmental effects were estimated by a statistical model that included contemporary group effect, type of kidding, genetic grouping, and kidding order. A multi-trait animal model with animal and permanent environment random effects was used to estimate genetic parameters and the significant environmental effects (fixed). Contemporary group influenced all traits; genetic grouping did not influence %LACT; type of kidding did not influence PFAT, %PROT or %LACT, and kidding order did not influence %FAT or %EXTR. Heritability and repeatability estimates were, respectively, 0.19 and 0.37 (MP270); 0.10 and 0.20 (PFAT); 0.12 and 0.24 (PPROT); 0.15 and 0.27 (PLACT); 0.13 and 0.24 (PEXTR); 0.21 and 0.34 (%FAT); 0.39 and 0.44 (%PROT); 0.17 and 0.29 (%LACT); 0.31 and 0.47 (%EXTR). Estimates of genetic correlations among MP270 and production of milk constituents were positive and high, but correlations between MP270 and %FAT, MP270 and %PROT, MP270 and %ESTR were moderate and negative. These heritability estimates show that satisfactory genetic gains can be obtained by selection, especially for milk constituents.

Effects of pentoxifylline during Schistosoma mansoni infection in Swiss mice: an analysis of worm burden, fecundity and liver histopathology.

The short-term effects of pentoxifylline (PTX) on granulomatous lesions during Schistosoma mansoni infection in Swiss mice were evaluated. Drug administration was initiated 42 and 140 days post-infection (DPI) for the acute and chronic infection groups, respectively. Treatment was carried out daily with 200 mg/kg (subcutaneous route) of the drug for five consecutive days. Recovery of parasites and tissues was performed at 49 DPI and 147 DPI, respectively. Liver histological analysis showed a decrease in the inflammatory reaction and fibrous content of the granulomas studied, and a significant reduction (P < 0.001) in their mean diameter was observed in the groups of rodents treated with PTX in acute and chronic infection, when compared to their respective control groups. However, no alteration in the number of S. mansoni recovered from the portal system was observed, and egg-laying kinetics was not notably modified by PTX treatment, and the immature stage distribution of S. mansoni eggs showed minor intrinsic variations with no statistical differences in the parameter second-stage/female/g among untreated mice and treated mice in acute and chronic infections, respectively, when evaluated by intestinal oograms. Data obtained indicate probable immunomodulatory effects of PTX in murine schistosomiasis both in acute and chronic infection.

Phoretic mites (Rhinoseius spp.) in Apodiformes from Cerrado and Pantanal Biomes in midwestern Brazil.

Nasal mites are commonly found in hummingbirds (Apodiformes). In most cases, endoparasitic mites were reported to parasitize the respiratory system, particularly the anterior nasal chambers, larynx, trachea, lungs, and the air and conjunctival sacs. However, some mites are only carried by hummingbirds, because flowers serving as a nutrient source for these small mites, such as for Apodiformes, a tropical nectar-feeding in their natural habitat. From August 2012 to May 2014, humminbirds were captured using mist nets, and examined for the presence of nasal mites (identified according to specific keys) in the Pantanal, and Cerrado biomes, from the Mato Grosso state, midwestern Brazil. Overall, 76 Apodiform specimens, from the Trochilidae family were captured, and 20 female nasal mites, identified as being from the Rhinoseius genus, were collected from eight hosts (10.5%), of the following species: Amazilia fimbriata, Amazilia versicolor, Eupetonema macroura, and Thalurania furcata. The Rhinoseius spp. has been reported on, in many ecosystems in the Americas, including Brazil, however, this is the first report on Apodiformes from the Pantanal biome. Phoretic mites are not responsible for direct damage to the population of Apodiformes, as they are not pathogenic. However, they compete for food with hummingbirds and decrease the quantity of food available in the environment. They also affect the reproductive dynamics of plants, feed on pollen, and interfere with the intricacies of the biodiversity where they live.

Effects of dietary supplementation with medicinal fungus in fasting glycemia levels of patients with colorectal cancer: a randomized, double-blind, placebo-controlled clinical study.

OBJECTIVE: The objective of this study was to evaluate the effects of dietary supplementation with Agaricus sylvaticus medicinal fungus in fasting glycemia of post-surgery patients with colorectal cancer. SCOPE: Proctology Ambulatory of Federal District Base Hospital-Brazil. SUBJECTS: Fifty-six colorectal cancer patients in postsurgery phase, stadiums phase I, II and II, which followed specific criteria of inclusion and exclusion. INTERVENTIONS: All patients were randomized in two groups: supplemented with Agaricus sylvaticus fungus (30 mg/kg/day) and placebo, and were followed up for six months. Subjects were divided later on according to BMI--Body Mass Index, sex and stage. Three fasting glycemia evaluations were carried out throughout the treatment. RESULTS: Subjects presented BMI medium = 24.65 kg/m2 (57.1% women and 42.9% men). The placebo group (average age 59.14 +/- 12.95 years) had initial glycemia levels of 94.36 +/- 15.34 mg/dL, after three months 98.12 +/- 15.54 mg/dL (p = 0.03) and in the sixth month 98.52 +/- 9.03 mg/dL (p = 0.01). The supplemented group (average age of 56.34 +/- 15.53 years) had initial glycemia levels of 95.92 +/- 11.64 mg/dL, after three months 94.88 +/- 12.24 mg/dL (p = 0.65) and, in the sixth month, a significant reduction to 92.86 +/- 6.82 mg/dL (p = 0.01). CONCLUSION: The results suggest that the dietary supplementation with Agaricus sylvaticus medicinal fungus can significantly reduce fasting glycemia levels of colorectal cancer patients in post-surgery phase.

Centrocestus formosanus (Trematoda: Heterophyidae) in Melanoides tuberculata (Gastropoda: Thiaridae) from Vila do Abraão, Ilha Grande, Rio de Janeiro, Brazil.

Pleurolophocercous cercariae found in the invasive gastropod Melanoides tuberculata (Müller, 1774) collected in a stream of the Vila do Abraão, Ilha Grande, Angra dos Reis, Rio de Janeiro, Brazil were used for experimental infection that enabled the identification of the heterophyid trematode Centrocestus formosanus (Nishigori, 1924). The parasite has been found in the locality since 2007, after two years of the introduction of M. tuberculata. Recently, from a sample of 483 specimens collected in June 2013, 101 (21%) were found infected with parasite. The potential environmental impacts caused by the parasite occurrence could be underestimated in the country, and actions to monitor and control both the parasite and the mollusk are necessary.

Experimental and molecular study of cercariae of Clinostomum sp. (Trematoda: Clinostomidae) from Biomphalaria spp. (Mollusca: Planorbidae) in Brazil.

Despite the large number of reports of species of Clinostomum from vertebrate hosts in South America, studies evaluating the molluscan transmitters of these parasites are scarce. In the present study, clinostomatoid cercariae shed from 0.02% (4/17,485) specimens of Biomphalaria spp., collected at the Pampulha reservoir, Belo Horizonte, Minas Gerais, Brazil, were used for experimental infection of Poecilia reticulata . Samples of cercariae from molluscs and metacercariae experimentally obtained from fish were subjected to morphological and molecular analyses and compared with species of Clinostomum reported in the Americas. The cercariae and metacercariae, here identified as Clinostomum sp., present general morphology similar to that reported for Clinostomum marginatum , however, from molecular point of view, differ significantly from North American C. marginatum and other species of Clinostomum reported in South America. These results suggest that the diversity of Clinostomum found in Brazil may be underestimated. Additional studies aimed at molecular characterization of South American species of Clinostomum, including the finding of specimens with sequences similar to that reported for C. marginatum in North America are required.

Phoretic mites (Rhinoseius spp. ) in Apodiformes from Cerrado and Pantanal Biomes in midwestern Brazil / Ácaros Foréticos (Rhinoseius spp. ) em Apodiformes dos Biomas Cerrado e Pantanal no Centro-Oeste do Brasil

Abstract Nasal mites are commonly found in hummingbirds (Apodiformes). In most cases, endoparasitic mites were reported to parasitize the respiratory system, particularly the anterior nasal chambers, larynx, trachea, lungs, and the air and conjunctival sacs. However, some mites are only carried by hummingbirds, because flowers serving as a nutrient source for these small mites, such as for Apodiformes, a tropical nectar-feeding in their natural habitat. From August 2012 to May 2014, humminbirds were captured using mist nets, and examined for the presence of nasal mites (identified according to specific keys) in the Pantanal, and Cerrado biomes, from the Mato Grosso state, midwestern Brazil. Overall, 76 Apodiform specimens, from the Trochilidae family were captured, and 20 female nasal mites, identified as being from the Rhinoseius genus, were collected from eight hosts (10.5%), of the following species: Amazilia fimbriata, Amazilia versicolor, Eupetonema macroura, and Thalurania furcata. The Rhinoseius spp. has been reported on, in many ecosystems in the Americas, including Brazil, however, this is the first report on Apodiformes from the Pantanal biome. Phoretic mites are not responsible for direct damage to the population of Apodiformes, as they are not pathogenic. However, they compete for food with hummingbirds and decrease the quantity of food available in the environment. They also affect the reproductive dynamics of plants, feed on pollen, and interfere with the intricacies of the biodiversity where they live.

Resumo Os ácaros nasais são comumente encontrados em beija-flores (Apodiformes). Na maioria dos casos, os ácaros foram relatados como endoparasitas, parasitando o sistema respiratório, particularmente as câmaras nasais anteriores, a laringe, a traqueia, os pulmões e os sacos aéreos e conjuntivais. No entanto, alguns ácaros são transportados apenas por beija-flores, pois flores servem como fonte de nutrientes para esses pequenos ácaros, assim como para Apodiformes tropicais alimentados de néctar em seu habitat natural. De agosto de 2012 a maio de 2014, beija-flores foram capturadas com redes de neblina e examinadas quanto à presença de ácaros nasais (identificados de acordo com chaves específicas) nos biomas Pantanal e Cerrado, no estado do Mato Grosso, Centro-Oeste do Brasil. No total, 76 exemplares Apodiformes da família Trochilidae foram capturados e 20 fêmeas de ácaros nasais, identificados como pertencentes ao gênero Rhinoseius, foram coletados de oito hospedeiros (10,5%), das seguintes espécies: Amazilia fimbriata, Amazilia versicolor, Eupetonema macroura e Thalurania furcata. Rhinoseius spp. foi relatado em muitos ecossistemas nas Américas, incluindo o Brasil, no entanto, este é o primeiro relato em Apodiformes do bioma Pantanal. Os ácaros foréticos não são responsáveis ​​por danos diretos à população de Apodiformes, pois não são patogênicos. No entanto, eles competem por comida com beija-flores e diminuem a quantidade de comida disponível no ambiente. Eles também afetam a dinâmica reprodutiva das plantas, se alimentam de pólen e interferem nas interações da biodiversidade em que vivem.

A method of decontaminating Strongyloides venezuelensis larvae for the study of strongyloidiasis in germ-free and conventional mice.

To study the possible influence of intestinal micro-organisms on the course of strongyloidiasis in mice, a method was developed to obtain axenic infective larvae of Strongyloides venezuelensis. Cultured larvae from conventional mice were treated with sodium hypochlorite 0.25% for 10 min, washed in distilled water and then exposed to various combinations of antibiotics for 30 or 60 min. Success was achieved with a combination of penicillin 180 mg/L and ceftazidime 1 mg/ml. Decontamination of the larvae was determined by aerobic and anaerobic culture and by inoculation into gnotobiotic mice. Viability was established by subcutaneous inoculation of larvae into germ-free and conventional mice. Preliminary results showed that gnotobiotic mice were more susceptible than conventional mice to infection with axenic S. venezuelensis larvae as judged by faecal egg excretion, recovery of worms in the small intestine and histopathological examination of the duodenal mucosa. These results suggest that the normal intestinal flora protects the host against experimental infection with S. venezuelensis.

In vivo cercaria-schistosomulum transformation: inhibitory effect of oxamniquine.

High doses of oxamniquine (given intramuscularly) produced inhibition of cercaria-schistosomulum transformation following intraperitoneal injections of cercariae into mice. Cercariae, tail-less cercarial bodies, and schistosomula were recovered from the peritoneal cavity of drug-treated mice in numbers significantly different from those recovered from untreated mice. Since untreated animals induced transformation of almost all the injected cercariae, the data suggest the compound is active during the process of host-larvae adaptation.

A marmoset model for some New World dermatotropic leishmaniases.

Callithrix penicillata is a good model for dermatotropic strains of some New World Leishmania species (L. mexicana mexicana, L. m. amazonensis, and L. braziliensis guyanensis). Both amastigotes and promastigotes readily infect these primates, with a relative short period of incubation. The developed lesions parallel those found in man, and despite the self healing presented by some marmosets, cell and serum immunological responses are presented. Skin alterations are confined to the site of inoculum, and visceralization is very rarely found. A good response to standard antimonial treatment makes the model useful for preclinical experiments with promising compounds.

Ultrasonographic criteria and tumor marker assay are good procedures for the diagnosis of ovarian neoplasia in preselected outpatients.

PURPOSE OF INVESTIGATION: To identify parameters for the diagnosis of ovarian neoplasia using ultrasonography (US) and serum tumor marker (TM: CA125, CA19.9, CA15.3, AFP, CEA and estradiol) assay. METHODS: Prospective study which included 373 women with increased ovarian volume (> 18 cm3 in premenopause and > 8 cm3 in postmenopause). US criteria (> or = 1) for surgery were: persistent (> 4 months) or increased cyst, cysts with > 1 thick septum or > or = 2 thin septa, cyst diameter > or = 7 cm, vegetation, calcification or cystic predominance (> 50%), solid tumor (> 50%). Doppler with a resistance index (RI) < 0.4 was considered abnormal. RESULTS: Laparotomy was performed in 164 (44%) patients with 66 (40.2%) benign neoplasias and 19 (11.6%) malignant cases (73.6% at Stage I or II). Two hundred and nine patients were maintained on clinical follow-up. The sensitivity for neoplasia and malignant neoplasia was, respectively, for RI: 17 and 63.6 and RI plus TM: 53.1 and 90.9. CONCLUSION: Ultrasound criteria and TM assay were indicated for the diagnosis of ovarian neoplasia.

Schistosoma mansoni: inflammatory foci around larvae in the peritoneal cavity of naive mice is radiosensitive.

Innate attack to Schistosoma mansoni cercariae was evaluated in irradiated mice. It was observed that 70% of the larvae from mice sacrificed one day after whole body irradiation with 400 or 800 rads were surrounded by cluster reactivities, without difference from controls. Differences were apparent on day 5 after irradiation with sub lethal (400 rads) or lethal doses (800 rads) suggesting that innate defence to infection take at least 5 days to be affected by low dose whole-body radiation.

Evaluation of the schistosomicidal efficacy of liposome-entrapped oxamniquine.

Oxamniquine (OXA) was successfully encapsulated in small unilamellar vesicles using a pH gradient method. This procedure led to a high drug encapsulation efficiency (> 85%) at a drug to lipid molar ratio of 1/10. Moreover, these liposomes were found to retain encapsulated OXA efficiently under dialysis conditions at 37 degrees C. Liposome-entrapped OXA (LOXA), OXA, and empty liposomes were tested against Schistosoma mansoni in a murine model. LOXA produced a significant reduction of the worm burden compared to the other preparations, when inoculated by subcutaneous route (s.c.) with 10 mg OXA/kg animal one day before the infection, and 3, 7, and 14 days after. However, LOXA was not effective when given 7 days before, or 35 days after infections. OXA, in the free form, was effective in relation to the untreated group, only when administered 3 days after the infection. Maximum effect of LOXA, with 97% reduction of the parasite number, was observed when the preparation was given s.c. one day before the infection. On the other hand, LOXA inoculated intraperitoneally one day before the infection didn't show any reduction of the parasite count. It can be concluded that LOXA is more effective than OXA for the treatment of experimental schistosomiasis, particularly when administered subcutaneously at a time close to the infection.

[Susceptibility of Biomphalaria tenagophila and Biomphalaria glabrata from a same region to 2 Schistosoma mansoni strains]. / Suscetibilidade de Biomphalaria tenagophila e Biomphalaria glabrata de uma mesma região a duas cepas de Schistosoma mansoni.

B. tenagophila snails from Ouro Branco, MG, showed positivity for S. mansoni, with infection rates of 5%, 10%, (SJ strain), and 1% (LE strain) using a pool of miracidia. The mollusks were found to be susceptive from the 3rd generation reared in laboratory onwards. The B. tenagophila (OB, MG) when individually exposed to 10 miracidia, showed infection rate of 2% for LE strain. B. glabrata snails from Gagé, MG, showed a positivity rate of 58% for S. mansoni (LE strain), under experimental conditions. The B. tenagophila from Cabo Frio, RJ and B. glabrata from Belo Horizonte, MG used as a control for SJ strain showed infection rates of 47%-85% and 36% respectivily. For the LE strain, B. glabrata (BH, MG) used as control showed infection rate of 40%-75%.