Colonização e lesão em fêmeas ingurgitadas do carrapato Rhipicephalus sanguineus causadas pelo fungo Metarhizium anisopliae

O presente trabalho teve como objetivo verificar a forma de penetração do fungo Metarhizium anisopliae [METSCH. (SOROKIN, 1883)] em carrapatos da espécie Rhipicephalus sanguineus (LATREILLE, 1806), assim como as lesões infringidas nos tecidos internos do ácaro. A forma de aderência e penetração do fungo foi estudada através da microscopia eletrônica de varredura e a ação do fungo nos tecidos internos avaliada em secções histológicas convencionais. Para observação destes eventos, realizaram-se infecções experimentais em 11 grupos de fêmeas ingurgitadas do carrapato R. sanguineus contendo 12 fêmeas ingurgitadas cada. Para tal, as fêmeas ingurgitadas foram banhadas durante 3 minutos, sob agitação manual, em suspensão com concentração 108 conídios/mL. No caso dos grupos controle o banho foi realizado apenas no veículo da suspensão. Os carrapatos foram processados para histopatologia e microscopia eletrônica em diversos tempos após a infecção, a saber: 1 e 18h, e um, dois, três, quatro, cinco, seis, sete, nove e onze dias. Observou-se que a maior parte dos conídios germinou em até 18h após a inoculação e que o fungo penetrou no ácaro através do tegumento 48h após a infecção. Após a penetração, o fungo invadiu o corpo do hospedeiro promovendo uma colonização difusa, sem preferência aparente por tecidos específicos. Dentre as lesões nos tecidos internos do ácaro, ressalta-se o rompimento da parede intestinal e vazamento do conteúdo para a hemocele. A morte do hospedeiro ocorreu entre 96 e 120h pós-infecção, e a esporulação do patógeno sobre o cadáver do ácaro iniciou-se em torno de 120 a 144h pós-infecção. Espera-se, com este trabalho, contribuir para o desenvolvimento e viabilização de técnicas de controle biológico dos carrapatos por fungos como alternativa ao uso de acaricidas.

Patogenicidade de isolados de Beauveria bassiana para ovos, larvas e ninfas ingurgitadas de Rhipicephalus sanguineus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Controle de larvas de Boophilus microplus por Metarhizium anisopliae em pastagens infestadas artificialmente

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Eventos externos e internos da infecção de larvas e ninfas de Rhipicephalus sanguineus por Metarhizium anisopliae

Examinaram-se a adesão, a germinação, a penetração e a colonização de larvas e ninfas de Rhipicephalus sanguineus por Metarhizium anisopliae, assim como as lesões infringidas pelo fungo nas respectivas fases do ciclo de vida do ácaro. Realizaram-se infecções experimentais em 11 grupos contendo 250 larvas e 11 grupos contendo 75 ninfas de R. sanguineus, por meio de banho, durante três minutos sob agitação manual, em suspensão contendo 10(8) conídios/ml do fungo. Nos grupos-controles, o banho foi realizado usando o veículo da suspensão. Larvas e ninfas foram processadas para um estudo histopatológico e de microscopia eletrônica de varredura nos seguintes tempos após a infecção: uma e 18 horas, e um, dois, três, quatro, cinco, seis, sete, nove e 11 dias. A germinação dos conídios ocorreu em até 18 horas pós-inoculação, e o fungo penetrou nas larvas e ninfas através do tegumento, dois e três dias após a infecção, respectivamente. Após penetração, o fungo invadiu o corpo das larvas e ninfas, promovendo uma colonização difusa, sem preferência aparente por tecidos específicos. Lesões significativas não foram observadas. A morte das larvas e ninfas ocorreu no terceiro e quarto dias pós-infecção, e a esporulação do patógeno sobre o cadáver foi iniciada no sexto dia pós-infecção.

In vitro acaricidal activity of neem (Azadirachta indica) seed extracts with known azadirachtin concentrations against Rhipicephalus microplus

The effect of four extracts from neem seeds (Azadirachta indica) containing 2000, 5000, 9000 and 10,000 ppm of azadirachtin A (AZA), quantified by high-performance liquid chromatography (HPLC) and diluted to 1.25%; 2.5%; 5.0%; 10.0% and 12.8% was verified by in vitro tests with engorged females and larvae of the cattle tick Rhipicephalus micro plus. The results from the bioassays with the engorged females showed that the main toxic effect of the extracts was reduction of the reproductive parameters, with a sharp drop in the number of eggs laid and the hatching rate, mainly when the extracts were diluted to 10.0% and 12.8%. The product effectiveness (PE) calculations for all the solutions tested showed that the AZA solution at 10,000 ppm (N10) was the most effective. However, statistical analysis of the PE data obtained for the proportional AZA concentrations in the different diluted extracts showed significance (P<0.05) of the effects included in the model (extract dilution, principle effect (classificatory) of the assay (extract) and the interaction between the two), indicating significant variations due to the dilution, the test and the interaction between the two factors in the tests with engorged females. For solutions N2, N5, and N9, it was not possible to estimate LC(90) values in the dilution range tested. The lowest LC(50) was observed for extract N5, and although extract N10 was the only extract for which the LC(90) could be estimated within the range tested, the LC(50) was higher than for N5 and N9. These results suggest that substances other than AZA present in the extracts influenced the efficacy, especially up to a certain LC range. In the tests with larvae, no mortality was observed, indicating zero effectiveness of all the extracts tested. The results of the tests with engorged females showed that the neem extracts had acaricide activity, inhibiting egg laying and the larval hatching rate. Complementary studies are necessary to develop new methods to isolate and/or identify other substances besides AZA contained in this plant, to enable using products made from it as acaricides. (C) 2011 Elsevier B.V. All rights reserved.

A Nonstaining and Tasteless Hydrophobic Salt of Chlorhexidine

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Overall-Mouth Disinfection by Photodynamic Therapy Using Curcumin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Association of ABO gene mutations resulting in a rare B subgroup

Background and Objectives B subgroups are rare and the genetic analysis reported to date has been limited.Materials and Methods Serological and molecular investigations were performed in blood from a B-subgroup donor.Results Red cells did not react with anti-B and anti-AB reagents. However, cells absorbed anti-B. Red cells presented positive reactions with anti-H, and saliva secreted H substance. The molecular study demonstrated a B allele with the substitutions 467C>T, 646T>A, 681G>A, 771C>T, 796C>A, 803G>C, 829G>A and an 0 allele with the sequence of 002.Conclusions It is probable that the presence in exon 7 of some of the 002 substitutions could have weakened the enzymatic activity of the encoded B transferase.

Eficácia do bochecho de quitosana a 0,4% sobre o biofilme e bactérias orais

The purpose of this study was evaluate the effectiveness of the chitosan at 0.4 with high molecular weight and high deacetylation degree mouthrinse over the total decrease of the streptococci, Streptococcus mutans, lactobaci/li and over the perceptible bacterial film and gingival bleeding indices. For that, a total of 68 healthy students between 11 and 13 years old, not allergic to crustacean and not users of antibiotics or antimicrobial agent for the last three months or during the treatment, was selected. From those, thirty two individuaIs used the mouthrinse test, and thirty six, the control one. The participants rinsed 10 mL of the solutions twice a day, one during the moming (which was supervised), and another one during the aftemoon (which was not supervised), for fifteen days. The saliva collect for the microbiological analysis, as well as the perceptible bacterial film and gingival bleeding indices check, were made before the use ofthe mouthrinses (base line), immediately after the last mouthrinse on the day (zero time) and fifteen days after (fifteen time). These data were collected at school and the saliva was carried inside the ice to the laboratory. The samples were diluted, and 0.1 mL ofthe 10 -1 dilution was seeded in Rogosa SL agar, for further analysis of the total of lactobaci/lus~ 0.1 mL of the 10-4 dilution in Mitis Salivarius with bacitracin, for S. mutans analysis; and 0.1 mL of the 10-6 dilution in Mitis Salivarius for the analysis ofthe total of streptococcus. The Rogosa SL agar plates were incubated in aerobic at 37°C for 72 hours and the MSB and the MS were incubated in anaerobic in Gaspak@ jars at 37°C for 48 hours for further count ofColonies Former Units (CFUs). The assay was made in duplicate for each bacterial group analyzed. The number of CFUs transformed in LOGlO was analyzed according to the following tests: ANOV A, t of Paired and Not Paired Student, Friedman, Man-Whitney and square-qui test. On the base line, alI the variables analyzed were similar on both tested groups. On both groups, for the total of streptococcus there was no significant difference along the time and for S. mutans there was a statistic significant increase of the CFUs from the base line to the zero time. For the total of lactobaccilus there was no significant difference on the test group along the time, and on the control there was a significant increase ofthe CFUs ITom the base line to the zero time. For both groups, there was significant decrease ofthe perceptible bacterial film index along the time, and that can be explained by the mechanic effect of the mouthrinse over the bacterial film and by the participation of the students on the research which could have motivated him to a better toothbrushing (Hawthome effect). The gingival bleeding index also showed a decrease along the time, even though it was not significant. Therefore, the conclusion of this study was that the chitosan at 0.4 % mouthrinse was not effective on the CFUs reduction of the three bacterial groups analyzed, as well as on the reduction of the perceptible bacterial film and gingival bleeding indices

Efeito in vitro da terapia fotodinâmica sobre bactérias orais em suspensão e na formação de biofilme : ensaio com azul de metileno e toluidina ativados por luz halógena

Photodynamic therapy (PDT) has been proposed as an alternative method for the treatment of biofilm-dependent oral diseases like dental caries. This therapy consists of simultaneous action of a visible light (L) and a photosensitizer (FS) in the presence of oxygen, which leads to production of different reactive oxygen species that can interact with the bacterial cell components, and promote cell death. This study aims to evaluate the antimicrobial action of PDT on oral bacteria in suspension, as well as the formation of mono and multi-species biofilms, in vitro, from a standard strain of Streptococcus mutans (ATCC 25175) and saliva samples, respectively. The dye methylene blue (MB) and toluidine blue (TB) were used at a concentration of 100 mg/ L and activated by halogen light (600 to 750 nm) from a modified hand held photopolymerizer (Ultralux ®, Dabi Atlante, Ribeirão Preto , São Paulo, Brazil.). Planktonic cultures were prepared and submitted to different experimental conditions: 1. PDT using TB 2. PDT using MB, 3. L+ FS- , 4. TB + L - ; 5. MB+ L-; 6. L- FS- (negative control) and 7. administration of 0.12% chlorhexidine digluconate (positive control) (Periogard ®, Colgate-Palmolive Company, New York, USA). The immediate and mediated action of PDT on bacterial suspensions, as well as its effect on biofilm formation were observed from the number of colony-forming units per milliliter (CFU/mL) and measures optical density (OD). The data were statistically analyzed using the Kruskal-Wallis and Mann-Whitney test for the significance level of 5%. According to the results, the PDT showed no antibacterial action on suspensions of S. mutans, regardless of the dye used. PDT with MB activated by halogen light was able to reduce 86.6% CFU/mL multi-species planktonic cultures, however, this reduction was not significant (p > 0.05). PDT showed antibacterial effect, mediate on multi-species planktonic cultures with TB (p < 0.001) and MB (p < 0.001), activated by halogen light. PDT was able to prevent the formation of multispecies biofilm, through the activation of TB by halogen light (p = 0.01). We conclude that activation of the dye toluidine blue and methylene blue, by halogen light (PDT) showed antimicrobial activity, compared to multi-species planktonic cultures prepared from saliva samples

Estudo das ligas titânio-zircônio resultantes do processo de fundição plasma-skull para aplicações como biomateriais

The aim of this work was to study a series of 11 different compositions of Ti-Zr binary alloys resistance to aggressive environment, i. e., their ability to keep their surface properties and mass when exposed to them as a way to evaluate their performance as biomaterials. The first stage was devoted to the fabrication of tablets from these alloys by Plasma-Skull casting method using a Discovery Plasma machine from EDG Equipamentos, Brazil. In a second stage, the chemical composition of each produced tablet was verified. In a third stage, the specimen were submitted to: as-cast microstructure analysis via optical and scanning electron microscopy (OM and SEM), x-ray dispersive system (EDS) chemical analysis via SEM, Vickers hardness tests for mechanical evaluation and corrosion resistence tests in a 0.9% NaCl solution to simulate exposition to human saliva monitored by open circuit potential and polarization curves. From the obtained results, it was possible to infer that specimens A1 (94,07 wt% Ti and 5,93% wt% Zr), A4 (77,81 wt % Ti and 22,19 wt % Zr) and A8 (27,83 wt% Ti and 72,17 wt% Zr), presented best performance regarding to corrosion resistance, homogeneity and hardness which are necessary issues for biomaterials to be applied as orthopedic and odontological prosthesis

Tropical dermatology: Venomous arthropods and human skin Part II. Diplopoda, Chilopoda, and Arachnida

Members of arthropod classes Chilopoda (centipedes), Diplopoda (millipedes), and Arachnida (spiders and scorpions) cause tissue injury via bites, stings, and/or a release of toxins. A few members of the Acari subclass of Arachnida (mites and ticks) can transmit a variety of infectious diseases, but this review will cover the noninfectious manifestations of these vectors. Dermatologists should be familiar with the injuries caused by these arthropods in order to initiate proper treatment and recommend effective preventative measures. (J Am Acad Dermatol 2012;67:347.e1-9.)

Ectopic ossification presenting as osteoid metaplasia in a salivary mucocele in a Shih Tzu dog

Background: Salivary mucocele is an accumulation of saliva in a single or multiloculated cavity lined by connective tissue that is contiguous to a salivary gland-duct complex and is the most common condition affecting the salivary glands in dogs. Occasionally, different types of metaplastic lesions, such as squamous and osseous metaplasia - which are rare lesions in animals - can be observed in association with salivary mucocele.Case presentation: A right facial enlargement was suddenly observed in a 4-year-old non-spayed female Shih-Tzu dog. The lesion presented itself as a soft and fluctuant mass located in the right side of the face near to the neck. Histologically, the mass consisted of a cavitary formation without an epithelial lining. Additionally, microscopic examination revealed the presence of osteoid-producing cells which gave rise to areas of bone formation, probably induced by irritation due to the presence sialoliths. Such cells and bone formations were also present in the cavity wall, consequently leading us to classify the condition as a salivary mucocele with osseous metaplasia.Conclusions: In the present case, the pathogenesis was probably associated with the presence of sialoliths, which can behave as etiological agents for the metaplastic lesion. The occurrence of osteoid metaplasia is a rare peculiar condition in the canine salivar y gland, and due to the rarity and lack of information about this specific disease, no clinical data can yet be associated with the development of salivary mucocele with osseous metaplasia in dogs.

Salivary cortisol to assess the hypothalamic-pituItary-adrenal axis in healthy children under 3 years old

Objective: To establish reference concentration intervals for salivary cortisol in healthy children, in the morning and in the afternoon, investigating factors that interfere with the concentration measured and the possibility that circadian rhythms are present.Methods: A controlled observational study was carried out with 91 children aged 45 days to 36 months, selected at random and living in Santo Andre, state of São Paulo, Brazil. Inclusion criteria were: healthy, well-nourished, free from fever and corticoid use, subdivided by age group (five subsets) at 6-month intervals. Saliva was collected during home visits in the morning and afternoon. Cortisol was radioimmunoassayed with cortisol 3-oxime-bovine albumin antiserum.Results: the five subsets exhibited higher cortisol concentration during the morning than in the afternoon (p < 0.001), and this difference passed 30% from 1 year of age onwards. Mean concentrations, in nmol/L, were 557.86 (morning) and 346.36 (afternoon). A negative linear correlation was observed between morning concentrations and hours' sleep and frequency of meals (p < 0,05), and in the afternoon with anthropometric measurements (p < 0.05).Conclusions: Reference values for normal salivary cortisol in healthy children were established. At:45 days it was possible to observe circadian rhythms, which reached maturity at 12 months of life. Sleep and food deprivation increased morning cortisol levels.

Prevalence of gastric lesions (ulcers and/or erosions) and their relationship to possible stressfull factors in asymptomatic Quarter Horse foals: endoscopic survey

A úlcera gástrica constitui-se numa das mais importantes causas de desconforto abdominal em eqüinos jovens. Com o objetivo de se verificar a prevalência de lesões gástricas (úlceras e/ou erosões) e sua relação com fatores como estresse, idade, e sexo, sessenta potros da raça Quarto de Milha não portadores de sinais clínicos compatíveis com lesões gástricas foram submetidos à gastroscopia. Os potros foram divididos em quatro faixas etárias de 15 animais cada uma, sendo: 1 a 30 dias, 31 a 60 dias, 61 a 90 dias e 91 a 120 dias de idade. A prevalência de lesões gástricas foi de 43,3%. Animais com idade entre 61 a noventa dias foram os mais acometidos. Não houve diferença significativa entre as faixas etárias. Descamações do epitélio aglandular ocorreram em nove potros (60%) entre um e trinta dias, em seis (40%) entre 31 e sessenta dias e em apenas dois (6,6%) com idade superior a sessenta dias. Fatores considerados estressantes, como infestação intensa por carrapatos (29), problemas respiratórios (3), dermatopatias (3), babesiose (2), onfaloflebite (1), diarréia (1), problemas ortopédicos (1) e ferida lacerante com presença de miíase (1) não influenciaram a ocorrência das lesões. Machos e fêmeas foram igualmente acometidos.