Infectious bronchitis virus replication in the chicken embryo related cell line

The susceptibility of the chicken embryo related (CER) cell line to infectious bronchitis virus (IBV M41) was characterized after five consecutive passages in CER cells. Virus replication was monitored by cytopathic effect observation, electron microscopy, indirect immunofluorescence, and reverse transcription polymerase chain reaction (RT-PCR). At 96 h post-infection (p.i.), the cytopathic effect was graded 75% by cell fusion, rounding up of cells and monolayer detachment, and the electron microscopy image characterized by coronavirus morphology. Cytoplasmic fluorescence was readily observed by from 24 h p.i. onwards, and at all times the respective viral RNA from IBV-infected monolayers was demonstrated by RT-PCR. Extra-cellular virus was measured by virus titration performed on chicken kidney cells and embryonated chicken eggs, and respective titres ranged from 4.0 to 6.0 log(10) EID50/ml on embryonated chicken eggs, and from 2.0 to 6.0 log(10) TCID50/ml on both CER cells and chicken kidney cells studied from 24 to 120 h p.i. These results confirmed that the M41 strain replicated well in the CER cell line.

Effect of lidocaine on the minimum alveolar concentration of sevoflurane in dogs

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

Índices de falha de transferência de imunidade passiva (FTIP) em bezerros holandeses e nelores, às 24 e 48 horas de vida: valores de proteína total, de gamaglobulina, de imunoglobulina G e da atividade sérica de gamaglutamiltransferase, para o diagnóstico de FTIP

Com o objetivo de determinar os índices de falha de transferência de imunidade em bezerros holandeses e nelores foram selecionadas, aleatoriamente, 413 amostras sanguíneas de animais de ambas as raças. Os filhos de vacas pluríparas e os bezerros holandeses apresentaram maiores níveis séricos de proteína total, da fração gamaglobulina e de IgG, do que bezerros da raça Nelore. Contudo, os índices de falha de transferência de imunidade passiva foram mais elevados nos animais da raça Holandesa, às 24 e 48 horas de idade. Estabeleceram-se valores de alguns componentes séricos para o diagnóstico de falha de transferência de imunidade passiva, de acordo com o desafio antigênico ambiental.

Cardiopulmonary and analgesic effects of caudal epidurally administered ropivacaine in cattle

Objective To assess cardiopulmonary and analgesic effects after administration of ropivacaine into the caudal epidural space of cattle.Study design Prospective, single-dose trial.Animals Eight healthy mixed breed cows aged 8 +/- 5 years and weighing 507 +/- 112 kg.Methods Caudal epidural anesthesia was produced in cows with 0.75% ropivacaine (0.11 mg kg(-1)). Onset time, duration and cranial spread of analgesia were recorded. Heart rate (HR), respiratory rate (f(R)), rectal temperature (RT), and mean arterial blood pressure (MAP) were measured prior to epidural administration (T-0) and at 15, 30, 60, 120, 180 and 240 minutes after epidural administration (T-15, T-30, T-60, T-120, T-180 and T-240). Arterial blood acid-base balance (pH, standard bicarbonate and base excess), gas tension (PaO2, PaCO2, SaO(2)) and electrolytes (Na+, K+, iCa(2+), Cl-) were recorded at T-0, T-30, T-60, T-120, T-180 and T-240. Ataxia was evaluated at T-0, T-30, T-60, T-120, T-180 and T-240 and at 1 hour intervals thereafter until analgesia was no longer present in each animal.Results Epidurally administered ropivacaine induced variable analgesia extending bilaterally from the coccyx to S3. Time to onset of analgesia and mean duration in the perineal area were 15 +/- 4 and 359 +/- 90 minutes, respectively. Respiratory rate and RT increased from T-120 to T-240 when compared to the value at T-0. Ionized calcium and chloride concentrations increased at T-180 and T-240 when compared to T-0. The other variables were not significantly different from baseline values (p > 0.05). Four animals were mildly ataxic.Conclusion and clinical relevance Ropivacaine (0.75%, 0.11 mg kg(-1)) can be administered by caudal epidural injection to produce prolonged bilateral perineal analgesia with minimal ataxia and cardiopulmonary changes in standing cattle.

Comparative Histopathological Analysis of Human Pulps after Class I Cavity Preparation with a High-Speed Air-Turbine Handpiece or Er:YAG Laser

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Marginal adaptation of Class 2 adhesive restorations

Objective: Control of microleakage represents a challenge for posterior composite restorations. The technique for composite placement may reduce microleakage. The null hypothesis of this in vitro study was that centripetal incremental insertion of composite resin would result in less microleakage than that obtained with the oblique incremental technique or bulk technique. Method and Materials: Standardized Class 2 preparations were made in 60 caries-free extracted third molars and randomly assigned to 3 groups ( n = 20): ( 1) oblique incremental insertion technique ( control), ( 2) centripetal incremental insertion technique, and ( 3) bulk insertion. The teeth were restored with a total-etch adhesive and micro-hybrid composite resin. The specimens were isolated with nail varnish except for a 2-mm-wide area around the restoration and then thermocycled ( 1,000 thermal cycles, 5 degrees C/ 55 degrees C; 30-second dwell time). The specimens were immersed in an aqueous solution of 50% silver nitrate for 24 hours, followed by 8 hours of immersion in a photo-developing solution and subsequently evaluated for leakage. The microleakage scores ( 0 to 4) obtained from the occlusal and cervical walls were analyzed with median nonparametric tests ( P <.05). Results: The null hypothesis was rejected. All techniques attained statistically similar dentin microleakage scores ( P =.15). The centripetal insertion technique displayed significantly less microleakage than the oblique technique at the enamel margins ( P =.04). Conclusion: None of the techniques eliminated marginal microleakage in Class 2 preparations. However, in occlusal areas, the centripetal technique performed significantly better than the other techniques.

Assessing the effects of vector control on dengue transmission

The dengue virus is an arbovirus transmitted to humans through mosquito Aedes aegypti. To describe the dynamics of dengue disease within these two populations we develop a compartment model taking into account chemical controls and mechanical control applied on the mosquitoes, which are the currently available controlling mechanisms to prevent dengue disease. To mimic seasonal variations, some parameters of the model are allowed to depend on time in order to divide a calendar year in favorable and unfavorable periods regarded to the development of vector population. (c) 2007 Elsevier B.V. All rights reserved.

Feeding habits of the spider crab Libinia spinosa H. Milne Edwards, 1834 (Decapoda, Brachyura) in Ubatuba Bay, São Paulo, Brazil

The main goal of this study was the identification of the items of the diet of the L. spinosa, based on the stomach contents analysis. The crabs were obtained from Ubatuba region north-eastern shore of São Paulo State. In the laboratory, all the individuals were dissected, the stomach was retreated and fixed in 10% formaline. The alimentary items were identified under stereomicroscope and analysed by the method of Frequency of Occurrence. A total of 194 stomachs was analysed and nine alimentary items were obtained. Unindentified material was found in 98% of analysed stomach and poriferan were present in less then 1% of stomachs. These results pointed a diversified diet explored by this crab, as well as the employment of some different methods for food intake. This suggested that these crabs could occupy different position in the trophic chain.

Disintegration of magnetic tablets in human stomach evaluated by alternate current Biosusceptometry

Oral administration is the most convenient route for drug therapy. The knowledge of the gastrointestinal transit and specific site for drug delivery is a prerequisite for development of dosage forms. The aim of this work was to demonstrate that is possible to monitor the disintegration process of film-coated magnetic tablets by multi-sensor alternate current Biosusceptometry (ACB) in vivo and in vitro. This method is based on the recording of signals produced by the magnetic tablet using a seven sensors array and signal-processing techniques. The disintegration was confirmed by signals analysis in healthy human volunteers' measurements and in vitro experiments. Results showed that ACB is efficient to characterize the disintegration of dosage forms in the stomach, being a research tool for the development of new pharmaceutical dosage forms. (C) 2003 Elsevier B.V. All rights reserved.

Production of pro- and anti-inflammatory cytokines by monocytes from patients with paracoccidioidomycosis

Monocytes and macrophages can produce a large repertoire of cytokines and participate in the pathogenesis of granulomatous diseases. We investigated the production of pro- and anti-inflammatory cytokines by monocytes from patients with active paracoccidioidomycosis. Peripheral blood monocytes from 37 patients and 29 healthy controls were cultivated with or without 10 mug/ml of lipopolysaccharide (LPS) for 18 h at 37 degreesC, and the cytokine levels were determined in the culture supernatants by enzyme immunoassay. The results showed that the endogenous levels of tumor necrosis factor alpha (TNF-alpha), interleukin-1 beta (IL-1beta), IL-6, IL-8, IL-10 and transforming growth factor beta detected in the supernatant of patient monocytes cultivated without stimulus were significantly higher than those produced by healthy controls. These data demonstrated that monocytes from patients with active paracoccidioidomycosis produce high levels of cytokines with both inflammatory and anti-inflammatory activities. However, patient monocytes produced significantly lower TNF-alpha and IL-6 levels in response to LPS when compared to normal subjects, suggesting an impairment in their capacity to produce these cytokines after LPS stimulation. Concentrations of IL-1beta, IL-8 and IL-10 in cultures stimulated with LPS were higher in patients than in controls. These results suggest that an imbalance in the production of pro- and anti-inflammatory cytokines might be associated with the pathogenesis of paracoccidioidomycosis. (C) 2003 Editions scientifiques et medicales Elsevier SAS. All rights reserved.

Biological properties of medicinal plants: a review of their antimicrobial activity

Plants have been used for thousands of years to flavor and conserve food, to treat health disorders and to prevent diseases including epidemics. The knowledge of their healing properties has been transmitted over the centuries within and among human communities. Active compounds produced during secondary vegetal metabolism are usually responsible for the biological properties of some plant species used throughout the globe for various purposes, including treatment of infectious diseases. Currently, data on the antimicrobial activity of numerous plants, so far considered empirical, have been scientifically confirmed, concomitantly with the increasing number of reports on pathogenic microorganisms resistant to antimicrobials. Products derived from plants may potentially control microbial growth in diverse situations and in the specific case of disease treatment, numerous studies have aimed to describe the chemical composition of these plant antimicrobials and the mechanisms involved in microbial growth inhibition, either separately or associated with conventional antimicrobials. Thus, in the present work, medicinal plants with emphasis on their antimicrobial properties are reviewed.

PCR detection of staphylococcal enterotoxin genes in Staphylococcus aureus strains isolated from raw and pasteurized milk

Milk is considered a nutritious food because it contains several important nutrients including proteins and vitamins. Conversely, it can be a vehicle for several pathogenic bacteria such as Staphylococcus aureus. This study aimed to analyze the frequency of genes encoding the staphylococcal enterotoxins (SEs) SEA, SEB, SEC, SED, SEE, SEG, SEH, SEI and SEJ in S. aureus strains isolated from raw or pasteurized bovine milk. S. aureus was found in 38 (70.4%) out of 54 raw milk samples at concentrations of up to 8.9 X 10(5) CFU/ml. This microorganism was present in eight samples of pasteurized milk before the expiration date and in 11 samples analyzed on the expiration date. of the 57 strains studied, 68.4% were positive for one or more genes encoding the enterotoxins, and 12 different genotypes were identified. The gene coding for enterotoxin A, sea, was the most frequent ( 16 strains, 41%), followed by sec (8 strains, 20.5%), sed (5 strains, 12.8%). seb (3 strains. 7.7%) and see (2 strains, 5.1%). Among the genes encoding the other enterotoxins, seg was the most frequently observed (11 strains. 28.2%), followed by sei (10 strains) and seh and sej (3 strains each). With the recent identification of new SEs, the perceived frequency of enterotoxigenic strains has increased. suggesting that the pathogenic potential of staphylococci may be higher than previously thoughts however, further studies are required to assess the expression of these new SEs by S. aureus. and their impact in foodborne disease. The quality of Brazilian milk is still low. and efforts from the government and the entire productive chain are required to attain consumer safety. (C) 2008 Elsevier B.V. All rights reserved.

Cytochemical characterization of the endomembranous system during the oocyte primary growth in Serrasalmus spilopleura (Teleostei, Characiformes, Characidae)

The morphophysiological changes that occur during oocyte primary growth in Serrasalmus spilopleura were studied using ultrastructural cytochemical techniques. In the previtellogenic oocytes endoplasmic reticulum components, Golgi complex cisternae and vesicles, lysosomes, multivesicular bodies and some electron-dense vesicles react to acid phosphatase (AcPase) detection. The endoplasmic reticulum components, Golgi complex cisternae and vesicles also react to osmium tetroxide and potassium iodide impregnation (KI). These structures, except for the Golgi complex cisternae, are strongly contrasted by osmium tetroxide and zinc iodide impregnation (ZIO). Some electron-dense vesicles are ZIO-stained, while microvesicles in the multivesicular bodies and other large isolated cytoplasmic vesicles are contrasted by KI. At primary oocyte growth, the activity of the endomembranous system and the proliferation of membranous organelles are intense. The biosynthetic pathway of the lysosomal proteins such as acid phosphatase, involves the endoplasmic reticulum, Golgi complex, vesicles with inactive hydrolytic enzymes and, finally, the lysosomes. The oocyte endomembranous system have reduction capacity and are involved in the metabolism of rich in SH groups. (c) 2005 Published by Elsevier Ltd.

Morphological regional differences of epithelial cells along the midgut in Diatraea saccharalis Fabricius (Lepidoptera : Crambidae) larvae

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

Prevalence of dog intestinal parasites and risk perception of zoonotic infection by dog owners in São Paulo State, Brazil

Coprological examination was used to estimate the prevalence of gastrointestinal parasites in stray and domiciled dogs from Botucatu, São Paulo State, Brazil. Risk factors for dog infection were assessed in relation to demographic, husbandry and management data. The dog owners completed a questionnaire survey on some aspects of dog parasitism such as parasite species, mechanisms of infection, awareness of zoonotic diseases and history of anthelmintic usage. Parasites were found in the faeces of 138 dogs, with an overall prevalence of 54.3%. Dogs harbouring one parasite were more common (31.4%) than those harbouring two (18.5%), three (3.2%) or four (1.2%). The following parasites and their respective frequencies were detected: Ancylostoma (37.8%), Giardia (16.9%), Toxocara canis (8.7%), Trichuris vulpis (7.1%), Dipylidium caninum (2.4%), Isospora (3.5%), Cryptosporidium (3.1%) and Sarcocystis (2.7%). Stray dogs were found more likely to be poliparasitized (P < 0.01) and presented higher prevalence of Ancylostoma, T. canis and Giardia (P < 0.01) than domiciled ones. Toxocara canis was detected more frequently in dogs with < 6 months of age (P < 0.05) and no effect of sex or breed could be observed (P > 0.05). Except for Ancylostoma, that showed a significantly higher prevalence in dogs living in a multi-dog household (P < 0.01), parasite prevalences were similar in single- and multi-dog household. The answers of dog owners to the questionnaire showed that the majority does not know the species of dog intestinal parasites, the mechanisms of transmission, the risk factors for zoonotic infections, and specific prophylactic measures. The predominance of zoonotic species in dogs in the studied region, associated with the elevated degree of misinformation of the owners, indicates that the risk of zoonotic infection by canine intestinal parasite may be high, even in one of the most developed regions of Brazil.