Effects of bovine leukemia virus infection on milk neutrophil function and the milk lymphocyte profile

The effects of bovine leukemia virus (BLV) on the immune response have been extensively investigated; however, its effects on mammary gland immunity are only speculative. Although BLV has a tropism for B cells, it can affect both adaptive and innate immunities because these systems share many effector mechanisms. This scenario is the basis of this investigation of the effects of BLV on mammary gland immunity, which is largely dependent upon neutrophilic functions. Thus, the present study sought to examine neutrophilic functions and the lymphocyte profile in the milk of naturally BLV-infected cows. The viability of the milk neutrophils and the percentage of milk neutrophils that produced reactive oxygen species (ROS) or phagocytosed Staphylococcus aureus were similar between BLV-infected and BLV-uninfected dairy cows. Furthermore, the expression of CD62L and CD11b by the milk neutrophils and the percentage of milk neutrophils (CH138⁺ cells) that were obtained from the udder quarters of the BLV-infected cows were not altered. Conversely, the median fluorescence intensity (MFI) representing intracellular ROS production and the phagocytosis of S. aureus, the expression of CD44 by the milk neutrophils and the percentage of apoptotic B cells were lower in the milk cells from BLV-infected dairy cows, particularly those from animals with persistent lymphocytosis (PL). The lymphocyte subsets were not different among the groups, with the exception of the percentage of CD5⁻/CD11b⁻ B cells, which was higher in the milk cells from BLV-infected cows, particularly those with PL. Thus, the present study provides novel insight into the implications of BLV infection for mammary gland immunity.     

Development and validation of a set of standard area diagrams to estimate severity of potato early blight

Standard area diagrams (SADs) to assess the severity of potato early blight (Alternaria grandis) on leaves of potato (Solanum tuberosum L.) were developed and validated. The proposed SADs include images of leaves with 10 distinct disease severities (0.1, 1, 3, 5, 10, 20, 40, 60, 80 and 100 %). The SADs were validated by 12 raters who had no previous experience in evaluating plant disease. Lin’s concordance correlation analysis of estimated vs. actual disease severity (based on image analysis) showed that precision and accuracy improved for most raters using the SADs, compared to assessments made without the SADs. The SADs improved accuracy (coefficient of bias, C _b ?=?0.97 and 0.99, without and with SADs, respectively) and agreement (Lin’s concordance correlation coefficient, ρ _c ?=?0.91 and 0.98 without and with SADs, respectively) of the estimates of severity. Severity estimates were more reliable when using SADs (coefficient of determination, R ² ?=?0.80 unaided and R ² ?=?0.95 with SADs, and the intra-class correlation ρ?=?0.86 without SADs and ρ?=?0.97 using the SADs). The SADs improved raters’ ability to accurately, precisely and reliably estimate potato early blight severity, and as such can be used to assess severity for several purposes, including breeding for resistance, fungicide screening, and pathotype characterization.     

RNAi‐based Bean golden mosaic virus‐resistant common bean (Embrapa 5.1) shows simple inheritance for both transgene and disease resistance

Bean golden mosaic virus (BGMV) is the causal agent of bean golden mosaic of common beans. A transgenic bean line that has been developed based on RNA interference to silence the BGMV rep gene showed immunity to the virus. Crosses were done between the transgenic line and six bean cultivars followed by four backcrosses to the commercial cultivars ‘Pérola’ and ‘BRS Pontal’. The transgene locus was consistently inherited from the crosses analysed in a Mendelian fashion in the segregating populations. The disease resistance reaction co‐inherited with the transgene. Nevertheless, the expression of disease resistance displayed a dosage effect phenomenon in the F₁ generation. The analysis of the homozygous near‐isogenic lines in field conditions, under high BGMV disease incidence, indicated that the transgenic lines were completely resistant. These results show the strength of the disease resistance obtained, the stability of the trait across generations and its usefulness in the management of a disease for which there is no reported Phaseolus germplasm with immunity.     

Efficacy of imiquimod 5% cream in the treatment of equine sarcoids: a pilot study

Imiquimod is an immune response modifier with potent antiviral and antitumour activity. The objective of this pilot study was to evaluate the efficacy of an imiquimod 5% cream (Aldaratrade mark: 3M, Saint Paul, MN, USA) as a topical treatment for equine sarcoids. Fifteen horses with a total of 19 tumours were enrolled, including mixed (7), fibroblastic (5), flat (3), verrucous (2), and nodular (2) types. Baseline data included history, physical examination, tumour location, measurement and digital photography. Imiquimod was applied by the owners three times a week until complete resolution of the tumour or 32 weeks, whichever occurred first. Tumours were measured and photographed every 4 weeks. Treatment efficacy was defined as 75% or greater reduction of tumour size by the end of the trial. Four sarcoids were withdrawn from the study. Twelve of the remaining 15 tumours (80%) showed more than 75% reduction in size and nine (60%) totally resolved between 8 and 32 weeks. The most common adverse effects of exudation, erythema, erosions, depigmentation and alopecia were limited to the tumour and adjacent areas. The results suggest that topical imiquimod is a therapeutic option for the treatment of equine sarcoids, although more detailed studies are required to corroborate these initial findings.     

Heart rate variability parameters of myxomatous mitral valve disease in dogs with and without heart failure obtained using 24-hour Holter electrocardiography

Time-domain heart rate variability (HRV) parameters and the correlation between echocardiography and Holter examinations in dogs with myxomatous mitral valve disease (MMVD) were determined. Holter examination was also performed at different time frames: an entire 24-hour period, a four-hour period during sleep, and a four-hour period while awake. Ten healthy (control group) and 28 MMVD dogs, 15 with and 13 without heart failure, were evaluated. The SDANN (sd of the mean normal RR intervals for all five-minute segments during 24-hour Holter) and pNN(50) (percentage of differences between adjacent normal RR intervals that are >50 ms computed over 24-hour Holter) variables were significantly lower in the dogs with MMVD heart failure. The differences in HRV between the groups were only detected during the 24-hour evaluation period (P<0.05). There were high correlations (canonical analysis) between Holter and echocardiography examinations when considering pNN(50), SDANN, and LA/AO (left atrial to aortic root ratio) (r=0.92; P<0.05), indicating that both are important in evaluating MMVD dogs. SDANN and pNN(50) are measures of parasympathetic control of the heart, and thus, it is possible to infer that the MMVD dogs exhibit parasympathetic withdrawal during the development of heart failure.     

Normalization of wood density in biomass estimates of Amazon forests

Wood density is an important variable in estimates of biomass and carbon flux in tropical regions. However, the Amazon region lacks large-scale wood-density datasets that employ a sampling methodology adequate for use in estimates of biomass and carbon emissions. Normalization of the available datasets is needed to avoid bias in estimates that combine previous studies of wood density that used wood sampling at diverse positions in the bole or with various methods of density determination. This paper examines the question of whether regressions for radial variation and for variation in wood density along the bole, both developed in dense forest in central Amazonia (CA), are suitable for the open forests in southern Amazonia (SA) that are currently the target of most of Amazonia's deforestation activity. The wood density of the heartwood and density of full disks or slices (bark, sapwood and heartwood) in each tree were measured to assess the radial variation. For variation along the length of the bole, wood densities at breast height and at the top of the bole were used. Moisture content of the bole was measured in SA and compared with values reported by studies from CA in similar dense forest. Comparing regressions that predict full-disk density from heartwood density, the pattern of radial variation differs slightly and significantly between the two forest types (ANCOVA p = 0.006); the slopes have similar values but the intercepts differ. Variation along the bole in the two forest types does not differ significantly (p = 0.144), so the CA model for predicting mean bole density from the density of a slice at breast height gives an unbiased estimate of the mean bole density when applied to SA trees. In SA the mean moisture content of the bole was 0.416 (±0.068 S.D.; n = 223 trees). Moisture content of the bole had a strong inverse relationship with basic wood density (r = −0.77), which explains the lower moisture content in the trees in CA relative to SA. A much weaker inverse relationship was found between moisture content and green wood density (r = −0.292). The relationship between wood basic density and green (‘fresh’) density presented in this study provides an alternative means of obtaining basic wood density directly in the field when oven drying of samples is not possible.     

Distinguishing sporulating and nonsporulating lesions as a method for evaluating the resistance of sugarcane genotypes to orange rust

Sugarcane breeding programmes rank the resistance of genotypes to Puccinia kuehnii, causal agent of orange rust, according to levels of disease severity. However, during the screening stages, this method of assessment can lead to precipitous elimination of genotypes with promising agronomic traits but showing mild symptoms of rust such as flecks or lesions that do not produce spores. This study aimed to propose a new method to classify the resistance of sugarcane genotypes to orange rust by counting sporulating lesions. Five sugarcane varieties with different levels of resistance to P. kuehnii were inoculated with two pathogen populations under controlled conditions. The disease severity (SEV), total number of lesions (TNL), and total number of sporulating lesions (TNSL) were evaluated in a 20 cm leaf fragment from the most diseased leaf. The TNL and TNSL evaluations were performed at 11, 16 and 21 days after inoculation (DAI) and SEV at 21 DAI. The thresholds of 80% and 8% of sporulating lesions (SL) separated susceptible from the intermediate varieties and intermediate from the resistant ones, respectively. It is proposed that the method of counting sporulating lesions be used in screening genotypes for resistance to P. kuehnii in sugarcane breeding programmes.