Genetic Diversity of the Cryptococcus gattii Species Complex in Mato Grosso State, Brazil.

Cryptococcosis is caused by fungi of the genus Cryptococcus. Owing to its importance, this study aimed to analyze the genetic diversity of C. gattii isolates from animals, humans, and the environment in Mato Grosso State (MT), Brazil, during November 2010-December 2017. All isolates of the C. gattii species complex were subjected to molecular genotyping via Restriction Fragment Length Polymorphism (PCR-RFLP) and Multi-locus Sequence Typing (MLST). PCR-RFLP analysis revealed that 21 isolates presented the genotype VGII, which is considered the most common and virulent genotype globally among. MLST analysis revealed the presence of 14 sequence types (STs), of which 5 are considered new genotypes. Clonal Complex (CC) CC182 (n = 5; 23,80%) and CC309 (n = 3; 14,28%) were the most frequent. CC distribution in relation to origin revealed that three CCs were found in animals with a predominance of CC182 (66,66%), while nine were found in humans, and two CCs were found in the environment. Extensive genetic variability was observed among the isolates in the State of Mato Grosso. STs belonging to the already described clonal complexes (CC) indicate the global expansion and adaptation of isolates in several other countries. Therefore, detection of clonal complexes and STs already described in other regions and the occurrence of new STs in the present study help further the current understanding of the geographic dispersion and genetic origin of the C. gattii species complex.

Cryptococcus gattii VGII in a Plathymenia reticulata hollow in Cuiabá, Mato Grosso, Brazil.

Little is known about the ecology of agents of cryptococcosis in Mato Grosso, without any data regarding to the sources of both agents in the environment. This study aimed to investigate Cryptococcus gattii and Cryptococcus neoformans associated with decay in tree hollows within the urban area of three different cities of Mato Grosso. Seventy-two environmental samples collected from 72 living trees in the cities of Cuiabá, Várzea Grande and Chapada dos Guimarães were sampled and analysed. One tree (Plathymenia reticulata, Leguminosae) in the city of Cuiabá yielded 19 colonies identified as C. gattii molecular type VGII. The isolation of C. gattii VGII in the downtown city of Cuiabá is important because it fits in the Northern Macroregion, suggesting expanding and urbanisation of this genotype in different Brazilian cities.

Implications of Mycoplasma hemofelis Bacterial Load on the Hematological Parameters of Naturally Infected Cats.

Mycoplasma hemofelis is the most pathogenic hemoplasma species that affect cats. M. hemofelis may cause an acute infection that leads to hemolytic anaemia. The objective of this study was to detect and to quantify the load of M. hemofelis in cats by conventional polymerase chain reaction (PCR) and by quantitative PCR (qPCR) and to describe the possible hematological changes. M. hemofelis DNA was detected in 28.6% of the randomly selected cats (42 of 147) attended at the Veterinary Hospital of the Federal University of Mato Grosso, Brazil. The agreement between conventional PCR and qPCR was substantive (k 0.6). Females were twice as likely to acquire infection as males (odds ratio, 2.31). There was no statistically significant association (P > .05) and little/no correlation between the hematological parameters and the average of bacterial load. The results indicate that M. hemofelis infection is not related to clinical signs and bacterial blood load in cats. The agreement between conventional and quantitative PCR made it possible to detect infection by M. hemofelis in a larger number of cats.

Molecular characterization of ovine zygomycosis in central western Brazil.

Zygomycosis is an important granulomatous disease that affects humans and animals, particularly sheep in tropical regions. Rhinofacial and nasopharyngeal zygomycosis were described in sheep in association with Conidiobolus spp. The present study characterized 5 samples of Conidiobolus isolated from 3 herds with clinical disease in Mato Grosso State, Brazil. The clinical and pathological findings were similar to nasopharyngeal zygomycosis. Based on morphological features, isolates were classified as Conidiobolus spp., and molecular phylogenetic analyses based on 18S ribosomal DNA grouped all isolates in a Conidiobolus lamprauges cluster. The current report describes the molecular characterization of ovine nasopharyngeal zygomycosis associated with C. lamprauges.

Identification of genes expressed by Cryptococcus gattii during iron deprivation.

Cryptococcus neoformans and C. gattii are pathogenic yeasts that cause life-threatening diseases in humans and animals. Iron is an essential nutrient for virtually every organism as it functions as a cofactor in numerous essential enzymatic reactions. In the literature, the competition for iron between microbes and mammalian hosts during infection is well documented. In this study, we used representational difference analysis (RDA) in order to gain a better understanding of how C. gattii responds to iron starvation. A total of 15 and 29 genes were identified as having altered expression levels due to iron depletion after 3 h and 12 h, respectively. Of these, eight genes were identified in both libraries. The transcripts were related to many biological processes, such as cell cycle, ergosterol metabolism, cell wall organization, transportation, translation, cell respiration and the stress response. These data suggest a remodeling of C. gattii metabolism during conditions of iron deprivation.

Molecular typing and in vitro antifungal susceptibility of Cryptococcus spp from patients in Midwest Brazil.

INTRODUCTION: Cryptococcosis is a systemic fungal infection that affects humans and animals, mainly due to Cryptococcus neoformans and Cryptococcus gattii. Following the epidemic of acquired immunodeficiency syndrome (AIDS), fungal infections by C. neoformans have become more common among immunocompromised patients. Cryptococcus gattii has primarily been isolated as a primary pathogen in healthy hosts and occurs endemically in northern and northeastern Brazil. We to perform genotypic characterization and determine the in vitro susceptibility profile to antifungal drugs of the Cryptococcus species complex isolated from HIV-positive and HIV-negative patients attended at university hospitals in Cuiabá, MT, in the Midwestern region of Brazil. METHODOLOGY: Micromorphological features, chemotyping with canavanine-glycine-bromothymol blue (CGB) agar and genotyping by URA5-RFLP were used to identify the species. The antifungal drugs tested were amphotericin B, fluconazole, flucytosine, itraconazole and voriconazole. Minimum inhibitory concentrations (MICs) were determined according to the CLSI methodology M27-A3. RESULTS: Analysis of samples yelded C. neoformans AFLP1/VNI (17/27, 63.0%) and C. gattii AFLP6/VGII (10/27, 37.0%). The MICs ranges for the antifungal drugs were: amphotericin B (0.5-1 mg/L), fluconazole (1-16 mg/L), flucytosine (1-16 mg/L), itraconazole (0.25-0.12 mg/L) and voriconazole (0.06-0.5 mg/L). Isolates of C. neoformans AFLP1/VNI were predominant in patients with HIV/AIDS, and C. gattii VGII in HIV-negative patients. The genotypes identified were susceptible to the antifungal drugs tested. CONCLUSION: It is worth emphasizing that AFLP6/VGII is a predominant genotype affecting HIV-negative individuals in Cuiabá. These findings serve as a guide concerning the molecular epidemiology of C. neoformans and C. gattii in the State of Mato Grosso.

In vitro susceptibility of Conidiobolus lamprauges recovered from sheep to antifungal agents.

Data regarding the susceptibility of Conidiobolus lamprauges is limited and there is no consensus about the optimal treatment for infections caused by Conidiobolus spp. In this context, the objective of this study was to evaluate the in vitro susceptibility of six C. lamprauges strains isolated from sheep conidiobolomycosis to amphotericin B, ketoconazole, fluconazole, itraconazole, posaconazole, voriconazole, anidulafungin, caspofungin, micafungin, flucytosine, and terbinafine using the CLSI M38-A2 microdilution technique. Terbinafine was the most active (MIC range <0.06-0.5 µg/mL). Resistance or reduced susceptibility was observed for amphotericin B and azole and echinocandin antifungals. Additional studies are necessary to determine the therapeutic potential of terbinafine as monotherapy or in combination therapy with other antifungals.

Natural infection of Leishmania (Viannia) braziliensis in Mus musculus captured in Mato Grosso, Brazil.

We report natural infection by Leishmania (Viannia) braziliensis in Mus musculus and Necromys lasiurus using molecular analyses (PCR-RFLP) of femoral bone marrow and skin fragments. The aim of this study was to detect infection by pathogenic species of Leishmania in small mammals in the state of Mato Grosso, Brazil. The animals were captured in Peixoto de Azevedo, a cutaneous leishmaniasis-endemic region located in the north of the state, from October 30 to November 3, 2008. Natural infection by Leishmania in synanthropic rodents may be a threat to humans due to cohabitation of human domiciles in this area.

Use of parasitological culture to detect Leishmania (Leishmania) chagasi in naturally infected dogs.

In Brazil, although the domestic dog is a major target for the control actions for visceral leishmaniasis, knowledge gaps of the Leishmania species present in those animals still exist in many endemic areas. The objective of this study was the use of parasitological culture as a diagnosis tool and identification of species of Leishmania and other trypanosomatids in the canine population in the city of Cuiaba/Mato Grosso. Biological samples such as blood, intact skin fragments, cutaneous ulcers, and bone marrow were collected during a cross-sectional study and cultured on biphasic medium (Novy-MacNeil-Nicolle [NNN]/Schneider's). Leishmania isolates were characterized through isoenzyme electrophoresis. Isolates were obtained from 11.2% (n=54) of the 482 animals studied considering the different anatomical sites investigated. Leishmania chagasi was confirmed in 8.3% (n=40) dogs and Trypanosoma caninum in 2.9% (n=14). The sample of intact skin presented a higher chance of isolation of L. chagasi in symptomatic dogs and bone marrow in asymptomatic dogs (p<0.05). The results presented in this study emphasize the value of culture and confirm, for the first time, the circulation of L. chagasi in the canine population in different neighborhoods of the city of Cuiaba and broaden the knowledge of the geographical distribution of T. caninum in Brazil.