Estrus cycle effect on muscle tyrosine kinase activity in bitches

Estrus cycle is a well recognized cause of insulin resistance in bitches. The insulin receptor (IR) as well as the insulin-like growth factor-I receptor belong to the same subfamily of tyrosine kinase (TK) receptors. The objective of this study was to evaluate basal TK activity in muscle tissue of bitches during the estrus cycle. Twenty-four bitches were used in the study (7 in anestrus, 7 in estrus, and 10 in diestrus). Muscle samples, taken after spaying surgery to determine TK activity, were immediately frozen in liquid nitrogen and then stored at −80°C until the membranes were prepared by sequential centrifugation after being homogenized. TK activity was determined by Poly (Glu 4:Tyr 1) phosphorylation and expressed in cpm/μg of protein. TK activity was significantly lower (P < 0.001) in the animals in estrus (104.5 ± 11.9 cpm/μg of protein) and diestrus (94.5 ± 16.9 cpm/μg of protein) when compared with bitches in anestrus (183.2 ± 39.2 cpm/μg of protein). These results demonstrate, for the first time, lower basal TK activity in the muscle tissue of female dogs during estrus and diestrus, which may represent lower insulin signaling capacity, opening a new field of investigation into the molecular mechanisms of insulin resistance in dogs.     

High throughput phenotyping for aphid resistance in large plant collections

ABSTRACT: BACKGROUND: Phloem-feeding insects are among the most devastating pests worldwide. They not only cause damage by feeding from the phloem, thereby depleting the plant from photo-assimilates, but also by vectoring viruses. Until now, the main way to prevent such problems is the frequent use of insecticides. Applying resistant varieties would be a more environmental friendly and sustainable solution. For this, resistant sources need to be identified first. Up to now there were no methods suitable for high throughput phenotyping of plant germplasm to identify sources of resistance towards phloem-feeding insects. RESULTS: In this paper we present a high throughput screening system to identify plants with an increased resistance against aphids. Its versatility is demonstrated using an Arabidopsis thaliana activation tag mutant line collection. This system consists of the green peach aphid Myzus persicae (Sulzer) and the circulative virus Turnip yellows virus (TuYV). In an initial screening, with one plant representing one mutant line, 13 virus-free mutant lines were identified by ELISA. Using seeds produced from these lines, the putative candidates were re-evaluated and characterized, resulting in nine lines with increased resistance towards the aphid. CONCLUSIONS: This M. persicae-TuYV screening system is an efficient, reliable and quick procedure to identify among thousands of mutated lines those resistant to aphids. In our study, nine mutant lines with increased resistance against the aphid were selected among 5160 mutant lines in just 5 months by one person. The system can be extended to other phloem-feeding insects and circulative viruses to identify insect resistant sources from several collections, including for example genebanks and artificially prepared mutant collections.     

The pre-emptive effect of epidural ketamine on wound sensitivity in horses tested by using von Frey filaments

Objective To evaluate the pre‐emptive analgesic effect of pre‐incisional epidural ketamine. Study Design A blinded, randomized experimental study. Animals Sixteen mixed breed mares, 7.6 ± 2.8 years old, weighing 352 ± 32 kg. Methods In a pilot study, an incision was made on one lateral thigh using a lidocaine block and no further analgesics, and it was verified that the nociceptive threshold was lower on the incised side than nonincised side (p ≤ 0.05), and that von Frey filaments evoked a pain response. The 16 animals were divided into group A (ketamine, n = 9) and B (saline, n = 7). An epidural catheter was inserted 24 hours before the trials. The thigh was shaved bilaterally, and the right side was blocked (incised side) using lidocaine. Twenty‐five minutes later, ketamine (A) or saline (B) was administered epidurally. Five minutes later, a 10‐cm skin incision was made on the right side, and then sutured. Nociceptive threshold was determined with von Frey filaments at 1, 3, and 5 cm around the incision at 15‐minute intervals for 2 hours, then at 4, 6, and 8 hours. Behavioral alterations, heart and respiratory rates were recorded. Nociceptive thresholds from these points were averaged to obtain mean values at each time, converted to a logarithmic scale, and submitted to a nonparametric analysis (Mann–Whitney and one‐way repeated measures anova test, p ≤ 0.05). Results After 8 hours, the global range score revealed reduced hyperalgesia (p < 0.01) around the incision in 92% (4.65–4.27) of evaluated intervals in group A (ketamine). There were no significant changes in behavior, heart and respiratory rates. Conclusions It was concluded that pre‐emptive epidural ketamine reduced post‐incisional pain in the horse, and that von Frey filaments were able to quantify cutaneous sensitivity after tissue damage. Clinical relevance Epidural ketamine injection can reduce post‐incisional sensitivity in the horse.     

A multiplex PCR for the detection of Brucella spp. and Leptospira spp. DNA from aborted bovine fetuses

Bovine brucellosis and leptospirosis are important causes of bovine abortion around the world. Both diseases can be serologically diagnosed, but many factors may cause false positive and negative results. Direct methods based on bacteriological isolation are usually employed, but they are difficult, time consuming and dangerous. Monoplex polymerase chain reaction (PCR) have been successfully described for the detection of Brucella spp. and Leptospira spp. Aiming at improvement in the direct diagnosis, a multiplex PCR (mPCR) for the detection of these agents in aborted bovine fetuses is described. The detection threshold of the mPCR was evaluated in experimentally contaminated bovine clinical samples using a conventional proteinase K/SDS or a boiling-based extraction protocols. The mPCR was applied to two groups of clinical samples: 63 episodes of bovine abortion and eight hamsters experimentally infected with Leptospira interrogans serovar pomona. Adopting microbiological isolation as reference, the test showed a sensitivity of 100% in both groups of clinical samples. Seven samples collected from bovine fetuses were Brucella spp. culture negative but showed positive results in mPCR. Regarding Leptospira spp. detection, similar results were observed in three bovine clinical samples. All hamsters infected with Leptospira were positive in both microbiological culture and mPCR. The boiling extraction protocol showed better results in some clinical samples, probably by the removal of PCR inhibitors by heat treatment. The high sensitivity, simplicity and the possibility of detection of both bacteria in a single tube reaction support the use of the mPCR described in the routine diagnosis.     

Determination of melamine content in amino resins by near-infrared spectroscopy

Melamine-urea-formaldehyde resin production has increased in the last years due to low formaldehyde emissions and improved moisture resistance when applied on wood-based panels. Melamine is a relatively high-cost raw material in the resin formulation and has a significant effect on wood-based panel properties. It is important for industry to implement efficient quality control methods to determine melamine content in resins. The main objective of this work was to develop chemometric techniques for off-line monitoring of melamine content using near-infrared spectroscopy. The use of interval partial least squares regression allowed identifying the most interesting spectral ranges for analysis (4,400–4,800 and 5,200–5,600 cm^?1) and the best preprocessing methods to use in the determination of this property. When the appropriate spectral range and preprocessing methods are selected, it is possible to obtain calibration models with high correlation coefficients.     

Presence of Leptospiral DNA in Semen Suggests Venereal Transmission in Horses

The purpose of the present study was to detect leptospiral DNA by PCR in semen and urine samples of stallions to test for venereal transmission in horses. A total of 10 stallions from four herds were studied, and sampling was conducted in semen and urine for culture and PCR and serum for serology. From the 10 serum samples tested, 6 (60%) were seroreactive. No pure culture was obtained, but leptospiral DNA was detected by PCR in 50% of the semen samples and 30% of urine samples. The present study aimed to detect leptospiral DNA by PCR in semen and urine samples of stallions to test for venereal transmission in horses. Based on these findings, we suggest that there is potential transmission of leptospirosis in horses by sexual transmission.     

Postpartum Variations of Plasma IGF and IGFBPs,Oocyte Production and Quality in Dairy Cows: Relationships With Parity and Subsequent Fertility

The aim of this study was to determine whether postpartum variations of plasma IGF‐1 and IGFBP concentrations, oocyte production and quality were related to parity and subsequent conception rate in Holstein dairy cows. Holstein dairy cows [10 primiparous (PP) and 22 multiparous (MP)] were allotted in six batches and sampled once weekly between calving and oestrous synchronization treatment started at 71.2 ± 2.0 days postpartum. During the 3 weeks before treatment, ovum pick‐up (OPU) was performed twice weekly. Oocytes were scored on a 4‐point scale, and oocytes from OPU1, 3 and 5 were fertilized in vitro. Seventeen cows became pregnant after first and second AI and were considered as fertile (F), while the others were considered to be subfertile (SF). Logistic regression was carried out to investigate the relationships between repeated measurements and fertility including parity and batch effects in the models. Likelihood of fertility significantly increased when plasma urea and IGFBP‐3 concentrations decreased and was higher in PP compared with MP cows. There was a trend for fertility to increase when plasma IGF‐1 concentrations increased (p = 0.07). In vitro cleavage and development rates were similar between SF and F cows (46.4% and 28.3% in SF vs 55.0% and 22.1% in F). Parity had an effect on plasma IGF‐1 concentrations (PP: 61.65 ± 2.67 vs MP: 41.63 ± 5.81 ng/ml, p < 0.001), mean number of follicles aspirated per session (PP: 5.7 ± 1.3 vs MP: 9.5 ± 0.8, p < 0.05) and fertility (PP: 8/10 = 80% vs MP: 9/22 = 41%, p < 0.05) but not on the number of oocytes recovered per session nor their quality. In conclusion, postpartum plasma urea and IGFBP‐3 concentrations, but not oocyte production and quality before breeding, were related to subsequent conception rate in our experimental design. Parity had a significant effect on energy status, follicular growth and fertility and needs to be considered when investigating relationships between nutrition and reproduction.     

Ozone Formation Potentials of Volatile Organic Compounds and Ozone Sensitivity to Their Emission in the Megacity of São Paulo, Brazil

In the present study, a three-dimensional Eulerian photochemical model was employed to estimate the impact that organic compounds have on tropospheric ozone formation in the Metropolitan Area of São Paulo (MASP). In the year 2000, base case simulations were conducted in two periods: August 22–24 and March 13–15. Based on the pollutant concentrations calculated by the model, the correlation coefficient relative to observations for ozone ranged from 0.91 to 0.93 in both periods. In the simulations employed to evaluate the ozone potential of individual VOCs, as well as the sensitivity of ozone to the VOC/NO _x emission ratio, the variation in anthropogenic emissions was estimated at 15% (according to tests performed previously variations of 15% were stable). Although there were significant differences between the two periods, ozone concentrations were found to be much more sensitive to VOCs than to NO _x in both periods and throughout the study domain. In addition, considering their individual rates of emission from vehicles, the species/classes that were most important for ozone formation were as follows: aromatics with a kOH?>?2?×?10⁴ ppm^?1 min^?1; olefins with a kOH?<?7?×?10⁴ ppm^?1 min^?1; olefins with a kOH?>?7?×?10⁴ ppm^?1 min^?1; ethene; and formaldehyde, which are the principal species related to the production, transport, storage and combustion of fossil fuels.     

Comparative analysis of Andiodrilus pachoensis casts in forests and pastures of South-Eastern Amazon (Brazil)

Soil ecosystem engineers produce biogenic structures (casts, mounds, galleries) that strongly affect soil processes. A comparative analysis of the physical characteristics of the casts produced by the earthworm Andiodrilus pachoensis was carried out in four pastures sowed with Brachiaria brizantha and four primary forests on the deforestation front of the Amazonian forest. In both systems, we recorded surface cast density and spatial patterns as well as surface cast physical properties. Cast and vegetation distributions were mapped within plots of sizes ranging from 25 to 50 m², and cast production was monitored during 2 months. Cast spatial distribution varied greatly among plots and across spatial scales ranging from regular at small spatial scales (0–20 cm) to clumped for larger distance ranges (>80 cm). Cast density was not significantly correlated with grass tuft density and their production was independent from the presence of grass tufts in pastures or litter quantity in forests. Although bulk soil properties (pH, C content, structural stability, etc) differed between pastures and forests, cast physical properties did not differ significantly among the studied ecosystem.