Development and validation of UV spectrophotometric method for orbifloxacin assay and dissolution studies

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

Soil physical and microbiological attributes cultivated with the common bean under two management systems

Agricultural management systems can alter the physical and biological soil quality, interfering with crop development. The objective of this study was to evaluate the physical and microbiological attributes of a Red Latosol, and its relationship to the biometric parameters of the common bean (Phaseolus vulgaris), irrigated and grown under two management systems (conventional tillage and direct seeding), in Campinas in the state of Sao Paulo, Brazil. The experimental design was of randomised blocks, with a split-plot arrangement for the management system and soil depth, analysed during the 2006/7 and 2007/8 harvest seasons, with 4 replications. The soil physical and microbiological attributes were evaluated at depths of 0.00-0.05, 0.05-0.10, 0.10-0.20 and 0.20-0.40 m. The following were determined for the crop: density, number of pods per plant, number of beans per pod, thousand seed weight, total weight of the shoots and harvest index. Direct seeding resulted in a lower soil physical quality at a depth of 0.00-0.05 m compared to conventional tillage, while the opposite occurred at a depth of 0.05-0.10 m. The direct seeding showed higher soil biological quality, mainly indicated by the microbial biomass nitrogen, basal respiration and metabolic quotient. The biometric parameters in the bean were higher under the direct seeding compared to conventional tillage.

Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immuno assay in horses from São Paulo State, Brazil

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

A simple high-performance liquid chromatography assay for on-line determination of catecholamines in adrenal gland by direct injection on an ISRP column

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

Evaluation of the double agar gel immunodiffusion test and of the enzyme-linked immunosorbent assay in the diagnosis and follow-up of patients with chronic pulmonary aspergillosis

Coordenação de Aperfeiçoamento do Pessoal de Nível Superior (CAPES)

Effectiveness of hyperbaric oxygen for experimental treatment of schistosomiasis mansoni using praziquantel-free and encapsulated into liposomes: Assay in adult worms and oviposition

The treatment of schistosomiasis depends on a single drug: praziquantel (PZQ). However, this treatment presents limitations such as low and/or erratic bioavailability that can contribute to cases of tolerance. Improvements to the available drug are urgently needed and studies with a controlled system of drug release, like liposomes, have been gaining prominence. The present study evaluated the activity and synergy between liposomal-praziquantel (lip.PZQ) and hyperbaric oxygen therapy (HBO). Mice received doses of 60 or 100mg/kg PZQ or lip.PZQ, 50 days post-infection, and after the treatment, were exposed to HBO (3 atmosphere absolute - ATA) for 1h. The viability of adult worms and oviposition were analyzed, by necropsy and Kato-Katz examination performed after 15 days of treatment. A concentration of 100mg/kg of lip.PZQ+HBO was more effective (48.0% reduction of worms, 83.3% reduction of eggs/gram of feces) and 100% of the mice had altered of oograms (indicating interruption of oviposition) compared to other treatments and to the Control group (infected and untreated). It is known that PZQ requires participation of the host immune system to complete its antischistosomal activity and that HBO is able to stimulate the immune system. The drug became more available in the body when incorporated into liposomes and, used with HBO, the HBO worked as an adjuvant. This explains the decreases of oviposition and worms recovered form hepatic portal system.

Improvement of an enzyme immunosorbent assay for detecting antibodies against Dioctophyma renale

An available enzyme-linked immunosorbent assay (ELISA) was studied for the detection of anti-Dioctophyma renale antibodies in the sera of dogs using, detection of parasite eggs in urine sediment as a reference test. ELISA uses a soluble antigenic preparation of esophagus of D. renale and the optimal dilutions of the antigen, serum and conjugate were determined by means of checker board titration, using positive (n=13) and negative (n=27) reference serum. The specificity and sensitivity of the ELISA were 93.8% and 92.3% respectively and the kappa index was good (0.76). These results suggest that ELISA described may prove to be an effective serological test for detecting dogs infected and exposed to this parasite mainly dogs that are not eliminating parasite eggs through their urine.

Validation of HPLC-UV assay of caffeic acid in emulsions

An accurate, sensitive, precise and rapid reversed-phase high-performance liquid chromatographic method was successfully developed and validated for the determination of caffeic acid (CA) in emulsions. The best separation was achieved on a 250 × 4.6 mm, 5.0 µm particle size RP18 XDB Waters column using ethanol and purified water (40:60 v/v) adjusted to pH 2.5 with acetic acid as the mobile phase at a flow rate of 0.7 mL/min. Ultraviolet detection was performed at 325 nm at ambient column temperature (25°C). The method was linear over the concentration range of 10-60 µg/mL (r(2) = 0.9999) with limits of detection and quantification of 1.44 and 4.38 µg/mL, respectively. CA was subjected to oxidation, acid, base and neutral degradation, as well as photolysis and heat as stress conditions. There were no interfering peaks at or near the retention time of CA. The method was applied to the determination of CA in standard and pharmaceutical products with excellent recoveries. The method is applicable in the quality control of CA.

Validation of absolute quantitative real-time PCR for the diagnosis of streptococcus agalactiae in fish

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

Chronic osteomyelitis of the maxilla and mandible: microbiological and clinical aspects

Osteomyelitis of the mandible and maxilla are common in developing countries and their treatment may be long-standing and difficult. Thus, the aim of this study was to discuss the main biological aspects of the chronic osteomyelitis of the jaws of especial interest for dental team. These infections are associated with a complex microbiota composed mainly by anaerobic bacteria, sometimes associated with microorganisms originated from the skin and digestive tract. These data suggest that chronic osteomyelitis of the mandible and maxilla should be treated as anaerobic infections in most cases. In addition, local surgical treatments are relevant in the therapy outcome, associated to the use of antimicrobial agents, and the failure to accomplish them is a major cause of treatment failure.

Pan-Assay interference compounds (PAINS): warning signs in biochemical-pharmacological evaluations

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

Influence of sanitization on the physico-chemical and microbiological quality of organic and conventional broccoli

The present study was carried out in order to evaluate the effect of chlorinated and ozonized water on the physico-chemical characteristics of broccoli, produced under organic and conventional cultivation procedures. Organic and conventional broccolis were subjected to two sanitation treatments, using chlorine and ozone, and were kept under cold storage for seven days. Analyses of pH, titrable acidity, soluble solids and weight loss were performed and the content of Cu, Mn and Zn was determined. In addition, the presence of pesticides was verified. The results show no influence of the cultivation method or the sanitation treatment on sample weight loss. Cultivation methods and sanitizing treatments affected broccoli pH, titratable acidity, and soluble solids during the evaluation period. No differences regarding the metal content on organic and conventional broccoli were observed. Furthermore, the presence of organochlorine compounds, nor other pesticides, was not detected both in organic and conventional vegetables.

Effect of natural antimicrobials on microbiological and sensorial quality of fresh-cut cantaloupe melon

The effect of different natural antimicrobials on the microbiological and sensorial quality of fresh-cut Cantaloupe melons stored up to 10 days at 5°C was examined. Pieces of melon were washed for 1 min at 5ºC in water (control), vanillin (1000 mg/L and 2000 mg/L) or cinnamic acid (148.16 mg/L and 296.32 mg/L). Other antimicrobial treatments consisted of packaging the pieces of melon with an antimicrobial pad which contained cinnamic acid (148.16 mg/L and 296.32 mg/L). After 10 days of storage, significant differences among antimicrobials treatments and water treatment were found. In water treatment, the psychrotroph load was 3.63 ± 0.09 log cfu g-1 meanwhile on all antimicrobial treatments the values ranged from 3.04 ±0.13 log cfu g-1 to 3.28±0.1 log cfu g-1. Mesophilic growth in the control treatment averaged 6.79±0.06 log cfu g-1 meanwhile on antimicrobial treatments the counts were from 5.15±0.01 log cfu g-1 to 5.30±0.03 log cfu g-1. Total coliform levels were 7.8±0.1 log cfu g-1 when melon was washed in water, followed by washing with cinnamon (296.32 mg/L) at 6.5 log cfu g-1 and for the rest of the treatments were around 5.5 log cfu g-1. The treatments did not display differences among mould and yeast growth after 10 days of storage. The sensorial quality decreased throughout storage. However, at the end of storage, the scores ranged between 6.5 and 7, above the minimum level for marketability (level 5). Sensorial panelist noted a ‘sweet’ taste when vanillin was used as sanitizer. In all antimicrobial treatments, no relation was found between a higher dose and a higher microbial reduction. So, vanillin at 1000 mg/L in water or cinnamic acid at 148.16 mg/L provided in water dip or as a pad inside the trays could be optimal natural sanitizers to substitute the use of chlorine in fresh-cut products as Cantaloupe melon.

Manejo de transição para agricultura orgânica, sob cultivo de citros (Citrus sinensis L. Osbeck), favorece a diversidade de fungos no solo

Pós-graduação em Ciências Biológicas (Microbiologia Aplicada) - IBRC

Animal-side serologic assay for rapid detection of Mycobacterium Bovis infection in multiple species of free-ranging wildlife

Numerous species of mammals are susceptible to Mycobacterium bovis, the causative agent of bovine tuberculosis (TB). Several wildlife hosts have emerged as reservoirs of M. bovis infection for domestic livestock in different countries. In the present study, blood samples were collected from Eurasian badgers (n = 1532), white-tailed deer (n = 463), brushtail possums (n = 129), and wild boar (n = 177) for evaluation of antibody responses to M. bovis infection by a lateral-flow rapid test (RT) and multiantigen print immunoassay (MAPIA). Magnitude of the antibody responses and antigen recognition patterns varied among the animals as determined by MAPIA; however, MPB83 was the most commonly recognized antigen for each host studied. Other seroreactive antigens included ESAT-6, CFP10, and MPB70. The agreement of the RT with culture results varied from 74% for possums to 81% for badgers to 90% for wild boar to 97% for white-tailed deer. Small numbers of wild boar and deer exposed to M. avium infection or paratuberculosis, respectively, did not cross-react in the RT, supporting the high specificity of the assay. In deer, whole blood samples reacted similarly to corresponding serum specimens (97% concordance), demonstrating the potential for field application. As previously demonstrated for badgers and deer, antibody responses to M. bovis infection in wild boar were positively associated with advanced disease. Together, these findings suggest that a rapid TB assay such as the RT may provide a useful screening tool for certain wildlife species that may be implicated in the maintenance and transmission of M. bovis infection to domestic livestock.