Molecular identification of Cryptosporidium spp. from fecal samples of felines, canines and bovines in the state of São Paulo, Brazil

The objective of this study was to obtain information of epidemiological nature through genotypic characterization of Cryptosporidium isolates from dogs, cats and bovines from the state of São Paulo, Brazil. The extraction of DNA from oocysts was carried out and polymerase chain reaction was accomplished using specific primers to 18S rRNA gene. The amplicons were directed sequenced. Seven cat samples, nine dog samples and nine bovine samples were analysed. From the seven cat samples the genotypic analyses revealed Cryptosporidium felis in all. These were the first genotypic characterization of Cryptosporidium from domestic felines in Brazil. In nine sequenced samples from dogs, genotypic identities compatible with Cryptosporidium canis were revealed in all samples. The genotypic analyses in bovines revealed Cryptosporidium parvum in eight samples and Cryptosporidium bovis in another sample, the last one being a non-zoonotic species, not related to clinical symptoms and described for the first time in Brazil. (c) 2007 Elsevier B.V. All rights reserved.

Comparison of conventional PCR, quantitative PCR, bacteriological culture and the Warthin Starry technique to detect Leptospira spp. in kidney and liver samples from naturally infected sheep from Brazil

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

Evaluation of a portable clinical analyzer for the determination of blood gas partial pressures, electrolyte concentrations, and hematocrit in venous blood samples collected from cattle, horses, and sheep

Objective To compare results reported for blood gas partial pressures, electrolyte concentrations, and Hct in venous blood samples collected from cattle, horses, and sheep and analyzed by use of a portable clinical analyzer (PCA) and reference analyzer (RA).Animals-Clinically normal animals (24 cattle, 22 horses, and 22 sheep).Procedures-pH; Pco(2); Po(2); total carbon dioxide concentration; oxygen saturation; base excess; concentrations of HCO(3)(-), Na(+), K(+), and ionized calcium; Hct; and hemoglobin concentration were determined with a PCA. Results were compared with those obtained for the same blood sample with an RA. Bias (mean difference) and variability (95% confidence interval) were determined for all data reported. Data were also subjected to analyses by Deming regression and Pearson correlation.Results-Analysis of Bland-Altman plots revealed good agreement between results obtained with the PCA and those obtained with the RA for pH and total carbon dioxide concentration in cattle, K(+) concentration in horses and sheep, and base excess in horses. Except for Na(+) concentration and Hct in horses and sheep, correlation was good or excellent for most variables reported.Conclusions and Clinical Relevance-Data from blood gas and electrolyte analyses obtained by use of the PCA can be used to evaluate the health status of cattle, horses, and sheep. Furthermore, the handheld PCA device may have a great advantage over the RA device as a result of the ability to analyze blood samples on farms that may be located far from urban centers. (Am J Vet Res 2010;71:515-521)

Molecular characterization of Cryptosporidium spp. from fecal samples of birds kept in captivity in Brazil

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

Histometric analysis of ligature-induced periodontitis in rats: a comparison of histological section planes

The purpose of this study was to analyze the histometry of ligature-induced periodontitis in rats at different histological section depths. Sixteen male adult Wistar rats were randomly assigned to two groups: ligature and control. In the ligature group, rats received a sterile 4/0 silk ligature around the maxillary right 2nd molar. Thirty serial sections containing the 1st and 2nd molars, in which the coronal and root pulp, cementoenamel junction (CEJ) in the mesial side of the 2nd molar, interproximal alveolar bone and connective fiber attachment were clearly visible, were selected for histometric analysis. The histological sections were clustered in groups of 10 sections corresponding the buccal (B), central (C) and lingual (L) regions of the of periodontal tissue samples. The distance between the CEJ in the mesial side of the 2nd molar and the attached periodontal ligament fibers (CEJ-PL) as well as the distance between the CEJ and the alveolar bone crest (CEJ-BC) were determined. From CEJ-PL and CEJ-BC distances measured for each specimen, the measurements obtained in the B, L and C regions were recorded individually and together. Data were submitted to statistical analysis. Significant differences (p<0.001) were observed between the control and ligature groups regarding CEJ-PL (0.05 mm and 0.26 mm, respectively) and CEJ-BC (0.47 mm and 0.77 mm, respectively) measurements. Regarding the depth of the buccal, central and lingual planes, the means of CEJ-PL and CEJ-BC of both groups showed no statistically significant differences (p>0.05). In conclusion, the selection of 10 serial sections of the central region of periodontal tissue samples at any depth can be considered as representative for the evaluation of periodontal ligament fiber attachment and bone loss in ligature-induced periodontitis in rats.

Influence of the type of dental trauma on the pulp vitality and the time elapsed until treatment: a study in patients aged 0-3 years

The purpose of the present study was to determine the influence of the type of trauma on the pulp vitality and the time elapsed until seeking dental care in children aged 0-3 years seen at the Baby Clinic of the Aracatuba Dental School, UNESP. A total of 1813 records were analyzed. Two hundred and three patients, corresponding to 302 traumatized teeth, were assessed clinically and radiographically. Hard-tissue injuries were the most frequent (52%), with a predominance of enamel crown fractures (41.4%), followed by concussions (12.6%) and intrusions (11.6%). Clinical and radiographic examination revealed that 72% of the traumatized teeth maintained pulp vitality. In the case of supporting-tissue lesions, 51.1% of the patients sought care within 1-15 days after injury, while in the case of hard-tissue injuries, 52.7% sought care only after 16 days. The results showed that supporting-tissue injuries had a significant influence on the faster seeking of dental care.

12-Month color stability of enamel, dentine, and enamel-dentine samples after bleaching

The study aimed to quantify the color regression of enamel (E), dentine (D), and combined enamel-dentine (ED) of differently bleached ED specimens over a period of 12 months in vitro. Two ED samples were obtained from the labial surfaces of bovine teeth and prepared to a standardized thickness with the enamel and dentine layer each 1 mm. The ED samples were distributed on four groups (each n=80), in which the different bleaching products were applied on enamel (1, Whitestrips; 2, Illumine 15%; 3, Opalescence Xtra Boost) or dentine surfaces (4, mixture of sodium perborate/distilled water). Eighty ED samples were not bleached (control). Color (L*a*b*) of ED was assessed at baseline, subsequently after bleaching and at 3, 6, and 12 months of storage after bleaching (each 20 samples/group). E and D samples were prepared by removing the dentine or enamel layer of ED samples to allow for separate color analysis. Bleaching resulted in a significant color change (Delta E) of ED specimens. Within the observation period, Delta L but not Delta b declined to baseline. L* values of E and D samples also declined and were not significantly different from control samples after 12 months, while b* values did not decrease to baseline. Generally, no differences between the bleaching agents could be observed. Color change of enamel, dentine, and combined ED of in vitro bleached tooth samples is not stable over time with regard to lightness. However, yellowness did not return to baseline within 1 year.

Influence of apical patency and filling material on healing process of dogs' teeth with vital pulp after root canal therapy

Foi propósito deste trabalho observar o processo de reparo de dentes de cães após obturação dos canais com dois cimentos diferentes, fazendo ou não a patência apical. Após uma sobreinstrumentação, os canais receberam um curativo de uma solução de corticosteróide-antibiótico por 7 dias, com o objetivo de obter invaginação de tecido conjuntivo para dentro dos canais. Após esse período, esse tecido foi totalmente removido em metade dos casos (grupos com patência apical) e preservados no restante dos casos (grupos sem patência apical). Os canais foram obturados pela técnica da condensação lateral empregando um cimento a base de hidróxido de cálcio (Sealer Plus) ou um cimento de Grossman (Fill Canal). Os animais foram sacrificados por overdose anestésica 60 dias após o tratamento endodôntico e as peças anatômicas foram obtidas e preparadas para análise histológica. Os dados obtidos foram analisados com base em diversos parâmetros histomorfológicos. Os resultados foram melhores nos grupos sem patência apical (p=0,01) do que nos grupos com patência. Dentre os cimentos estudados, os melhores resultados foram observados com o cimento Sealer Plus (p=0,01). em conclusão, tanto a patência apical (presença ou ausência) quanto o tipo de material obturador de canal influíram no processo de reparo apical de dentes de cães com polpas vitais após tratamento endodôntico. O emprego de um cimento a base de hidróxido de cálcio em dentes sem patência apical promoveu os melhores resultados, dentre as condições experimentais propostas.

Evaluation of mineral trioxide aggregate and calcium hydroxide cement as pulp-capping agents in human teeth

This study evaluated the histomorphologic response of human dental pulps capped with mineral trioxide aggregate (MTA) and Ca(OH)(2) cement (CH). Pulp exposures were performed on the occlusal floor of 40 human permanent premolars. After that, the pulp was capped either with CH or MTA and restored with composite resin. After 30 and 60 days, teeth were extracted and processed for histologic exam and categorized in a histologic score system. The data were subjected to Kruskal-Wallis and Conover tests (alpha = .05). All groups performed well in terms of hard tissue bridge formation, inflammatory response, and other pulpal findings. However, a lower response of CH30 was observed for the dentin bridge formation, when compared with MTA30 and MTA60 groups. Although the pulp healing with calcium hydroxide was slower than that of MTA, both materials were successful for pulp capping in human teeth.

Direct detection of infectious bursal disease virus from clinical samples by in situ reverse transcriptase-linked polymerase chain reaction

The presence of the very virulent (vv) Brazilian strain of infectious bursal disease virus (IBDV) was determined in the bursa of Fabricius, thymus and liver of 2-week-old broilers from a flock with a higher than expected mortality. For this purpose, a direct in situ reverse transcriptase (RT)-linked polymerase chain reaction (PCR) method was developed using specific primers for vvIBDV. Unlabelled forward and reverse biotinylated oligonucleotides were used for RT-PCR in a one-step method and the respective products were revealed by a direct enzymatic reaction. The results were compared with those obtained by standard RT-PCR using general primers for IBDV and virus isolation. The virus isolation, RT-PCR and in situ RT-PCR revealed positive results on the bursa of Fabricius in 86%, 80% and 100%, respectively. The in situ RT-PCR detected vvIBDV in all tested thymus and liver samples, whereas the standard RT-PCR detected virus in 80% and 90% of the samples, respectively. After three consecutive passages on chicken embryonated eggs, IBDV was isolated from 64% of the thymus samples and 30% of the liver samples. In the present study, no classical or antigenic variants of IBDV were detected. The developed in situ RT-PCR assay was able to detect the very virulent strain of IBDV with a higher sensitivity than the conventional RT-PCR and virus isolation.

Short-term evaluation of the pulpo-dentin complex response to a resin-modified glass-ionomer cement and a bonding agent applied in deep cavities

Objectives. To evaluate the response of the pulpo-dentin complex following application of a resin-modified glass-ionomer cement or an adhesive system in deep cavities performed in human teeth.Methods. Deep class V cavities were prepared on the buccal surface of 26 premolars. In Group I the cavity walls (dentin) and enamel were conditioned with 32% phosphoric acid and the dentin adhesive system One Step (Bisco, Inc., Itasca, IL, USA) was applied. In Groups 2 and 3, before total etching and application of bonding agent, the cavity floor was lined with the resin-modified glass-ionomer cement-Vitrebond (3M ESPE Dental Products Division, St. Paul, MN, USA) or the calcium hydroxide cement-Dycal (control group, Dentsply, Mildford, DE, USA), respectively. The cavities were restored using light-cured Z-100 composite resin (3M ESPE). The teeth were extracted between 5 and 30 days and prepared for microscopic assessment. Serial sections were stained with H/E, Masson's trichrome, and Brown and Brenn techniques.Results. In Group 1, the inflammatory response was more evident than in Groups 2 and 3. Diffusion of dental material components across dentinal tubules was observed only in Group 1, in which the intensity of the pulp response increased as the remaining dentin thickness decreased. Bacteria were evidenced in the lateral walls of two samples (Group 2) which exhibited no inflammatory response or tissue disorganization.Conclusions. Based on the experimental conditions, it was concluded total acid etching followed by application of One Step bonding agent cannot be recommended as adequate procedures. In this clinical condition the cavity walls should be lined with a biocompatible dental material, such as Vitrebond or Dycal. 2003 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Human pulp response after an adhesive system application in deep cavities

Objectives: To evaluate the pulpo-dentin complex response to a dentin adhesive application in deep cavities performed in human teeth.Methods: Deep class V cavities were prepared on the buccal surface of 46 premolars. The remaining dentin of the axial wall received 10% phosphoric acid and dentin adhesive (group DA), or was protected before the acid and dentin adhesive application with calcium hydroxide cement (group CH). Half of the teeth, which received the acid application directly over the axial wall, were contaminated prior to the procedures with dental plaque collected from the patient's own teeth (group DAC). The plaque was placed on the dentin for 5 min and then the cavity was washed. All teeth were restored with a light-cured composite resin. The teeth were extracted after 7, 30 or 60 days and prepared according to normal histologic techniques. Serial sections were stained with WE, Masson's trichrome and Brown & Brenn technique for demonstration of bacteria.Results: the histopathologic evaluation showed that in groups DA and DAC, the inflammatory response was more evident than in group CH. Also, the intensity of the pulp reaction increased as the remaining dentin thickness decreased. There was no statistical difference in the inflammatory response between the groups DA and DAC.Conclusion: Based on the experimental conditions, we concluded that the All Bond 2 adhesive system, when applied on dentin in deep cavities, showed an acceptable biocompatibility. However, the intensity of the pulpo-dentin complex response depends on the remaining dentin thickness. (C) 1999 Elsevier B.V. Ltd. All rights reserved.

Biocompatibility of resin-based materials used as pulp-capping agents

Aim To evaluate and compare the response of pulps of rats capped with resin-modified glass-ionomer cement (RMGIC) or self-etching adhesive system.Methodology Class I cavities were prepared on the occlusal surface of 54 maxillary first molars of 27 rats. Pulp exposure was performed on the cavity floor. The following resin-based materials were applied as pulp-capping agents: G1, Clearfil Liner Bond 2V (CLB 2V; Kuraray Co., Japan); G2, Vitrebond (VIT; 3M/ESPE, USA). In group 3 (control group), a calcium hydroxide/saline paste (CH; Labsynth, Brazil) was used. The cavities were restored with amalgam. After 7, 30 and 60 days, the animals were sacrificed and the jaws were processed for microscopic evaluation.Results Despite the inflammatory response caused by the experimental and the control materials at 7 days, pulpal healing associated with calcified barrier formation was observed at 60 days following the pulp therapy. Both resin-based materials promoted a large zone of cell-rich fibrodentine matrix deposition on the pulp horn related to the pulp exposure site, which was larger to VIT than to CLB 2V specimens. Tertiary dentine underneath the fibrodentine matrix was deposited by a layer of elongated pulpal cells. The remaining pulpal tissue exhibited normal histological characteristics. In the control group, healing and dentine-bridge formation was observed at 30 days. Pulpal breakdown occurred only when bacterial infection occurred.Conclusion Both experimental pulp-capping agents allowed pulpal healing characterized by cell-rich fibrodentine and tertiary dentine deposition as well as calcified barrier formation.

Clinical and microbiological performance of resin-modified glass-ionomer liners after incomplete dentine caries removal

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

Human pulp responses to in-office tooth bleaching

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