Optimization and validation of analytical conditions for cholesterol and cholesterol oxides extraction in chicken meat using response surface methodology

The analytical conditions for the extraction of cholesterol and cholesterol oxides in chicken meat were optimized by means of response surface methodology. The separation and identification were performed by normal phase HPLC using UV and refractive index (RI) detectors, and the confirmation of the 11 cholesterol oxides identities in the samples was verified by HPLC-APCI-MS. The developed methodology showed good analytical performance, presenting recovery levels from 84 to 103% and detection limits varying from 0.01 to 0.06 microg/g for UV detection and from 1.98 to 2.12 microg/g for RI detection. The present study demonstrated the presence of 22 R-hydroxycholesterol, 24 S-hydroxycholesterol, and 22 S-hydroxycholesterol for the first time in chicken meat.     

Pathogenic and molecular comparison of Puccinia kuehnii isolates and reactions of sugarcane varieties to orange rust

Sugarcane orange rust, a disease caused by Puccinia kuehnii, was first reported in Brazil in 2009. There are no studies comparing the Brazilian P. kuehnii collections and the reaction of important sugarcane varieties under controlled conditions. This work compared the reaction of seven sugarcane varieties inoculated with six different P. kuehnii isolates from Brazilian sugarcane areas and verified the pathogenic and genetic variability of these isolates. The incubation (I) and latency (L) disease periods, disease severity (SEV), total number of lesions (TNL), total number of sporulating lesions (TNSL), and percentage of sporulating lesions (%SL) were evaluated. Furthermore, ITS1 and IGS ribosomal sequences of all P. kuehnii isolates used in this study were compared with pathogen sequences from 13 different countries. The disease incubation ranged from 7 to 10 days and the latency ranged from 10 to 21 days. SEV and TNL showed large variations and few significant differences between the reaction of the varieties to P. kuehnii, in contrast with the variables TNSL and %SL. The P. kuehnii isolates did not compose different virulent races, but the isolate from one site (Araras) was a more aggressive race. The ITS1 and IGS ribosomal sequences of six P. kuehnii isolates were identical with each other and to most P. kuehnii American sequences deposited at GenBank. The studied sequences of P. kuehnii isolates differed from the sequences from Asia, Tahiti and Oceania.     

Arterial pulse wave propagation velocity in healthy dogs by pulse wave Doppler ultrasound

The aim of this study was to prospectively evaluate the carotid-femoral pulse wave velocity (PWV) values in healthy dogs using pulse wave Doppler ultrasound. A secondary aim was to determine the feasibility of this method and to report the intra- and interobserver reproducibilities of the PWV in conscious dogs. The data were studied in 30 healthy, adult, male (n?=?15) and female (n?=?15) dogs. The time interval marked between the R wave peak of the electrocardiogram and the intersection of the blood flow wave upstroke of the Doppler spectrum with the baseline of zero frequency was determined for the carotid (T₁) and for the femoral (T₂) arteries. The distance covered by the pulse wave (L) was determined. The PWV was then calculated using the following formula: L/T₂ - T₁. The mean values of PWV calculated from the total sample (n?=?30) evaluated were 13.41?±?2.20 m/s. No significant statistical difference was observed for the PWV measurements between males (14.82?±?3.18 m/s) and females (12.64?±?2.45 m/s). The analysis revealed no intra nor interobserver differences. A reasonable reproducibility of the PWV measurements was showed by intraclass correlation coefficients (ICC), and the coefficients of variation (CV). These data demonstrate that noninvasive vascular Doppler analysis is a feasible and reproducible method to determine the carotid-femoral PWV in dogs.     

Detection of Bemisia tabaci Mediterranean cryptic species on soybean in São Paulo and Paraná States (Brazil) and interaction of cowpea mild mottle virus with whiteflies

Since the detection of Bemisia tabaci Middle East-Asia Minor 1 (MEAM1) species in Brazil in the early 1990s, this whitefly has been the predominant species and the main viral vector in the country. In this work, whiteflies were collected from commercial soybean fields near and far from greenhouses where Mediterranean (MED) species had previously been detected infesting vegetable crops. Results indicated that MEAM1 was the predominant whitefly species in soybean in most sampling sites, while MED was found colonizing soybean plants in open field conditions alone and/or with MEAM1 in several places. Among the tested insects, MED species was mostly detected harbouring the facultative endosymbiont Hamiltonella. We also detected cowpea mild mottle virus (CPMMV) infecting soybean plants. Transmission assays demonstrated that MED species was more efficient transmitting CPMMV from beans to beans and from soybean to soybean plants than MEAM1, while MEAM1 was more efficient in transmitting the virus from soybean to bean than MED. Performance assays indicated that adult emergence of both species on CPMMV-infected soybean plants was higher when compared with the emergence on healthy plants. Moreover, nonviruliferous MED and MEAM1 adults preferred to settle more often on healthy plants, while viruliferous adults settled more often on CPMMV-infected soybean plants. As MED has already been found in soybean open fields in São Paulo and Paraná States, Brazil, and it is a good vector of CPMMV, we anticipate problems related to whitefly management and to increase in the incidence of the virus in soybean.