Mário Paulo Amante Penatti

Prevalence of intestinal parasites in preschool children in the region of Uberlândia, State of Minas Gerais, Brazil

INTRODUCTION Children are an important high-risk group for helminth and protozoa infections. Daycare centers are environments where children have proven to be more susceptible to acquiring intestinal parasites. Thus, the purpose of this study was to verify the prevalence of intestinal parasites in children who attended the two daycare centers maintained by the local government of Uberlândia, State of Minas Gerais, Brazil. METHODS Fecal samples were collected from 133 children (73 children at the Public Preschool for Early Childhood Education, PPECE A, and 60 at the PPECE B) following identification according to sex and age and agreement to participate by parents or guardians who signed the free, informed consent form. The samples were examined by the Lutz method. RESULTS Coproparasitological tests performed on 133 children showed that 29.3% of them were parasitized for enteroparasites or commensals, 6.7% of the children presented polyparasitism. Among the protozoa, Giardia lamblia were the most prevalent and Hymenolepis nana were the most frequent among the helminths. CONCLUSIONS Thus, analysis of the results showed that intestinal parasites still represent a public health problem, especially among children and in areas where the socioeconomic and educational conditions are less favorable.

Escherichia coli strains from edema disease: O serogroups‚ and genes for Shiga toxin‚ enterotoxins‚ and F18 fimbriae

The objectives of the research were to determine the presence of the gene sequences for Shiga Toxin 2e (Stx2e), enterotoxins (ST-I, ST-II and LT-I), and F18 fimbriae in 144 Escherichia coli strains isolated from pigs with edema disease; to assess the ability of stx2e(+) strains to produce Stx2e; and to determine the O serogroups of the E. coli strains. Presence of the genes was determined by polymerase chain reaction (PCR), production of Stx2e was assessed by cytotoxicity for Vero and Hela cells, O serogroups were identified by agglutination with specific antisera. Of the 144 strains tested, 99 were stx2e(+) by PCR, but only 45 of these were Stx2e(+) in the cell culture assays. Among the 99 stx2e(+) strains, PCR detected the genes for F18ab, ST-I, ST-II, LT-I in 76, 40, 31 and 16 strains, respectively. Forty-one of the 99 sxt2e(+) strains belonged to O group 139; the rest did not belong to the classical edema disease O serogroups. It is likely that the enterotoxins, whose genes were detected at high frequency, are responsible for diarrhea seem in pigs with edema disease in Brazil.

Micro-leakage at the implant-abutment interface with different tightening torques in vitro

Objectives This study evaluated the microleakage at the implant/abutment interface of external hexagon (EH) implants and abutments with different amounts of bacteria and tightening torques. Material and Methods A bacterial suspension was prepared to inoculate the implants. The first phase of this study used nine EH implants and abutments that were divided into three groups with different amounts of bacterial suspension (n=3): V0.5: 0.5 µL; V1.0: 1.0 µL e V1.5: 1.5 µL, and tightened to the manufacturers recommended torque. The second phase of this experiment used 27 assemblies that were similar to those used in the first phase. These samples were inoculated with 0.5 µL of bacterial suspension and divided into three groups (n=9). T10: 10 Ncm; T20: 20 Ncm and T32: 32 Ncm. The samples were evaluated according to the turbidity of the broth every 24 hours for 14 days, and the bacteria viability was tested after that period. The statistical evaluation was conducted by Kruskal-Wallis testing (p<.05). Results During the first phase, groups V1.0 and V1.5 was presented with bacterial contamination in all samples after 24 h. During the second phase, two samples from group T10 and one from T20 presented positive results for bacterial contamination. Different amounts of bacterial solution led to overflow and contamination during the first 24 h of the experiment. The tightening torques did not statistically affect the microleakage in the assemblies. However, the group that was tightened to 32 Ncm torque did not show any bacterial contamination. Conclusion After 14 days of experimentation, the bacteria were proven to remain viable inside the implant internal cavity.

Enzymatic and hemolytic activity in different Candida species

BACKGROUND Candida species, in conditions of microbiota imbalance or decreased immune defenses, may be one of the main human fungal pathogens. Virulence factors constitute the mechanisms used by the fungus to avoid host defenses. AIMS This study aimed to investigate the in vitro production of virulence factors, such as hemolytic activity, and deoxyribonuclease (DNase), proteinase, and phospholipase activities in Candida spp. METHODS Fifty clinical isolates were analyzed for virulence factors: Candida albicans (15), Candida tropicalis (15), Candida parapsilosis (10), Candida glabrata (5), and Candida krusei (5). Hemolytic activity was determined in Sabouraud dextrose agar plates containing 3% glucose and 7% sheep red cells. Culture media containing, respectively, agar-base DNA, egg yolk, and bovine albumin were used to determine DNase, phospholipase and proteinase activities, respectively. RESULTS Forty-eight (96%) of 50 isolates showed hemolytic activity, with 10 (20%) positive for DNase, 19 (38%) for proteinase, and 16 (32%) for phospholipase. Statistically significant differences were observed between species for phospholipase (p<0.0001) and proteinase (p<0.05) production. CONCLUSIONS It is concluded that all species had hemolytic activity. DNase activity was detected in all species except in C. glabrata; proteinase activity was detected in C. albicans, C. tropicalis, and C. parapsilosis; and phospholipase activity was observed in C. albicans and C. tropicalis.

Different culture media containing methyldopa for melanin production by Cryptococcus species

INTRODUCTION Melanin production by species of Cryptococcus is widely used to characterize C. neoformans complex in mycology laboratories. This study aims to test the efficacy of methyldopa from pharmaceutical tablet as a substrate for melanin production, to compare the production of melanin using different agar base added with methyldopa, and to compare the melanin produced in those media with that produced in Niger seed agar and sunflower seed agar by C. neoformans, C. laurentii, and C. albidus. Two isolates of each species, C. neoformans, C. laurentii, and C. albidus, and one of Candida albicans were used to experimentally detect conditions for melanin production. METHODS The following media were tested: Mueller-Hinton agar (MHA), brain and heart infusion agar (BHIA), blood agar base (BAB), and minimal medium agar (MMA), all added with methyldopa, and the media Niger seed agar (NSA) and sunflower seed agar (SSA). RESULTS All isolates grew in most of the culture media after 24h. Strains planted on media BAB and BHIA showed growth only after 48h. All isolates produced melanin in MMA, MHA, SSA, and NSA media. CONCLUSIONS Methyldopa in the form pharmaceutical tablet can be used as a substrate for melanin production by Cryptococcus species; minimal medium plus methyldopa was more efficient than the BAB, MHA, and BHIA in the melanin production; and NSA and SSA, followed by MMA added with methyldopa, were more efficient than other media studied for melanin production by all strains studied.

FREQUENCY OF Candida SPECIES IN A TERTIARY CARE HOSPITAL IN TRIANGULO MINEIRO, MINAS GERAIS STATE, BRAZIL

Infections by Candida species are a high-impact problem in public health due to their wide incidence in hospitalized patients. The goal of this study was to evaluate frequency, susceptibility to antifungals, and genetic polymorphism of Candida species isolated from clinical specimens of hospitalized patients. The Candida isolates included in this study were obtained from blood cultures, abdominal fluids, and central venous catheters (CVC) of hospitalized patients at the Clinical Hospital of the Federal University of Uberlândia during the period of July 2010 - June 2011. Susceptibility tests were conducted by the broth microdilution method. The RAPD-PCR tests used employed initiator oligonucleotides OPA09, OPB11, and OPE06. Of the 63 Candida isolates, 18 (28.5%) were C. albicans, 20 (31.7%) were C. parapsilosis complex species, 14 (22.2%) C. tropicalis, four (6.4%) C. glabrata, four (6.4%) C. krusei, two (3.3%) C. kefyr, and one (1.6%) C. lusitaniae. In vitro resistance to amphotericin B was observed in 12.7% of isolates. In vitroresistance to azoles was not detected, except for C. krusei. The two primers, OPA09 and OPB11, were able to distinguish different species. Isolates of C. albicans and C. parapsilosis complex species presented six and five clusters, respectively, with the OPA09 marker by RAPD-PCR, showing the genetic variability of the isolates of those species. It was concluded that members of the C. parapsilosis complex were the most frequent species found, and most isolates were susceptible to the antifungals amphotericin B, flucozanole, and itraconazole. High genetic polymorphisms were observed for isolates of C. albicans and C. parapsilosis complex species, mainly with the OPA09 marker.

Phenotypic Characteristics of Yeasts of the Genus Candida and Cryptococcus in Differential Culture Media

In the clinical mycology laboratory, the identification of yeast species is done by screening in specific media, such as chromogenic agar for Candida species and Niger seed agar for Cryptococcus species, both of which are of clinical interest. This study aimed to evaluate the growth and morphological characteristics of yeasts of the Candida and Cryptococcus species in different culture media. Yeast species included in the study were: C. albicans, C. dubliniensis, C. glabrata, C. tropicalis, C. krusei, C. lipolytica, C. parapsilosis, C. metapsilosis, C. orthopsilosis, C. neoformans, C. gattii, C. flavescens, and C. albidus. The media used were Sabouraud dextrose agar, Sabouraud dextrose broth, hypertonic Sabouraud broth (plus 6.5% NaCl), Candida chromogenic agar, methyldopa agar, Niger seed agar and tobacco agar. Growth, color, size, presence of fringes, melanin and the appearance of the colony were evaluated. All isolates grew in the media used, except for the hypertonic Sabouraud broth; in Candida chromogenic agar, C. albicans and C. dubliniensis present a green color and C. tropicalis a blue color, while other species show colors including pink, purple, gray and white; in the Niger seed agar, C. neoformans, C. gattii and C. flavescens presented a brown color, while others had white colonies; in tobacco agar, the colors included white, cream and gray; and in methyldopa agar, all colonies were white. Some isolates presented colonies with fringes in the tobacco, methyldopa and Niger seed agar; the presence of melanin was observed by Cryptococcus isolates in the Niger seed and tobacco agar; the appearance of colonies in the media varied from opaque to shiny or mucoid, according to the isolate and the culture medium. All of the culture media used allowed the growth of the tested isolates, except for C. lipolytica, which did not grow in hypertonic Sabouraud broth. The isolates of Cryptococcus, C. krusei and C. dubliniensis presented a significant reduction of growth in hypertonic Sabouraud broth. K e y w o r d s

Occurrence of F42 colonization factor in Escherichia coli strains isolated from piglets with diarrhea

The objective of this study was to determine the presence of the colonization factor F42 in 168 strains of Escherichia coli isolated from diarrheic stools of newborn piglets. The presence of F42 in 12 (7.1%) strains was detected with the agglutination test. Through the Polymerase Chain Reaction (PCR) of F42 positive strains, gene encoding enterotoxins (ST-I, ST-II, LT-I and LT-II) were detected. The finding of ST-I/ST-II genes in 50% of the strains, ST-I (16%) and ST-II (25%) indicates a strong association of FC F42 with heat-stable enterotoxins (91%). In contrast, the thermolabile enterotoxin (LT-I and LT-II) genes were not detected. Serogroups of F42 positive strains were determined, serogroup O8 being the most prevalent (41,7%). Other serogroups, as there are O9, O11, O18, O32, O35, O98 and O101, were also identified. Thus, FC F42 was confirmed as an additional factor of virulence in the pathogenesis of porcine colibacillosis.