Crop beta-glucanase activity limits the effectiveness of a recombinant cellulase used to supplement a barley-based feed for free-range broilers

1. The supplementation of diets rich in soluble polysaccharides with microbial cellulases and hemicellulases decreases digesta viscosity and promotes broiler performance. 2. In contrast, recent experiments suggest that polysaccharidases are ineffective for improving the nutritive value of pasture biomass used by free-range broilers. However, the feasibility of using cellulases and hemicellulases to improve the utilisation of cereal-based feeds by pastured poultry remains to be established. 3. A study was undertaken to investigate the capacity of a recombinant cellulase from Clostridium thermocellum to improve the nutritive value of a barley-based feed for free-range pastured broilers of the RedBro Cou Nu x RedBro M genotype. 4. The results show that supplementation of a barley-based diet with a recombinant beta-glucanase had no effect on the performance of free-range broilers, foraging in legume-based diets from d 28 to 56. In addition, the results confirm that the lack of effect of the recombinant enzyme in improving the nutritive value of the barley-based feed does not result from enzyme proteolysis or inhibition in the gastrointestinal tract. 5. Significantly, beta-glucanase activity was identified in the crop of non-supplemented animals. The data suggest that endogenous cellulases originated both from the barley-based feed and from the crop microflora. 6. The results presented here suggest that in older birds of slow-growing genotypes associated with free-range production systems, previously unknown sources of beta-glucanases, such as the feed and microbial symbiotic microflora, can affect the effectiveness of exogenous enzymes added to the feed.     

Development and validation of a standard area diagram set to aid assessment of severity of loquat scab on fruit

A standard area diagram set (SAD) to aid visual assessment of loquat scab (caused by Fusicladium eriobotryae) severity on fruit was developed and evaluated for improving accuracy, precision and reliability of visual estimates. The SAD set contains eight black and white diagrams of diseased fruit with severity values from 2 % to 98 %. To evaluate the SADs, a group of 20 raters (comprising 10 ‘experienced’ and 10 ‘inexperienced’ raters) assessed the same set of 50 images three times, the first without SADs and the second and third using the SADs as an aid. Only for the group of inexperienced raters did SADs significantly improve accuracy (bias correction factor, C _b?=?0.93 without SADs and 0.98 with SADs), precision (correlation coefficient, r?=?0.88 without SADs and r?=?0.96 with SADs) and overall agreement (Lin’s concordance correlation coefficient, ρ _c?=?0.82 without SADs and ρc with SADs = 0.95) of the estimates. Accuracy and precision of the estimates by inexperienced raters were significantly higher than those obtained by the experienced raters, especially for the second assessment with SADs. Inter-rater reliability was improved when SADs were used by inexperienced raters, whereas a high degree of intra-rater reliability was obtained by both experienced and inexperienced raters when using SADs. The SADs developed in this study were useful for obtaining more accurate, precise and reliable assessments of loquat scab for inexperienced raters, and should be used as an aid for assessing scab in epidemiological studies or monitoring for decision-making purposes.     

Species composition and genetic structure of Fusarium graminearum species complex populations affecting the main barley growing regions of South America

Members of the Fusarium graminearum species complex (FGSC), such as F. graminearum and F. asiaticum, are the main cause of fusarium head blight (FHB) of wheat and barley worldwide. In this study, 117 FGSC isolates obtained from commercial barley grain produced in Argentina (n = 43 isolates), Brazil (n = 35), and Uruguay (n = 39) were identified to species and trichothecene genotypes, and analysed using amplified fragment length polymorphism (AFLP) and sequence‐related amplified polymorphism (SRAP) markers. In addition, reductase (RED) and trichothecene 3‐O‐acetyltransferase (Tri101) were sequenced for a subset of 24 isolates. The majority of the isolates (n = 103) were identified as F. graminearum, which was the only species found in Argentina. In Uruguay, only one F. cortaderiae isolate was found among F. graminearum isolates. In Brazil, F. graminearum also dominated the collection (22/35), followed by F. meridionale (8/35), F. asiaticum (2/35), F. cortaderiae (2/35) and F. austroamericanum (1/35). Species were structured by trichothecene genotype: all F. graminearum were of the 15‐acetyldeoxynivalenol (ADON), F. meridionale, F. asiaticum and F. cortaderiae were of the nivalenol (NIV), and F. austroamericanum was of the 3‐ADON genotype. Both AFLP and SRAP data showed high levels of genetic variability, which was higher within than among countries. Isolates were not structured by country of origin. SRAP analysis grouped F. graminearum in a separate cluster from the other species within the complex. However, AFLP analysis failed to resolve the species into distinct clades with partial clustering of F. meridionale, F. austroamericanum, F. asiaticum and F. graminearum isolates.     

A two-enzyme constituted mixture to improve the degradation of Arthrospira platensis microalga cell wall for monogastric diets

The main goal of this study was to test a rational combination of pre-selected carbohydrate-active enzymes (CAZymes) and sulphatases, individually or in combination, in order to evaluate its capacity to disrupt Arthrospira platensis cell wall, allowing the release of its valuable nutritional bioactive compounds. By the end, a two-enzyme constituted mixture (Mix), composed by a lysozyme and a α-amylase, was incubated with A. platensis suspension. The microalga cell wall disruption was evaluated through the amount of reducing sugars released from the cell wall complemented with the oligosaccharide profile by HPLC. An increase of the amount of reducing sugars up to 2.42 g/L in microalgae treated with the Mix relative to no treatment (p < .05), as well as a 7-fold increase of oligosaccharides amount (p < .001), were obtained. With resort of fluorescence microscopy, a 36% reduction of fluorescence intensity (p < .001) was observed using Calcofluor White staining. In the supernatant, the Mix caused a 1.34-fold increase in protein content (p = .018) relative to the control. Similarly, n-6 polyunsaturated fatty acids (PUFA) (p = .007), in particular 18:2n-6 (p = .016), monounsaturated fatty acids (MUFA) (p = .049) and chlorophyll a (p = .025) contents were higher in the supernatant of microalgae treated with the enzyme mixture in relation to the control. Taken together, these results point towards the disclosure of a novel two-enzyme mixture able to partial degrade A. platensis cell wall, improving its nutrients bioavailability for monogastric diets with the cost-effective advantage use of microalgae in animal feed industry.     

Profitability of fungicide applications for managing soybean rust in scenarios of variable efficacy and costs: A stochastic simulation

Soybean rust (SBR) in Brazil is controlled with fungicides, which have shown variable, sometimes declining, efficacy over time. A Monte Carlo simulation framework was implemented as a decision tool and to estimate the probability for a fungicide programme being profitable depending on efficacy and total cost defined by the user. Probability distributions were fitted to slopes and intercepts of the disease–yield relationship and severity in the untreated plots reported in the literature, as well as historical records of soybean price. Simulations of disease reduction conditioned to predefined control efficacy and total application costs were split into scenarios that combined two categories of severity (high and low) and two attainable yield classes (high and low). These categories were defined based on the median of severity (57.8%) and median of the intercept (yield when severity is zero, 2995.1 kg/ha). Probability matrices were constructed relating fungicide efficacy and costs. A higher frequency of break-even events occurred in scenarios of high disease pressure and higher yield. Yearly simulations, starting with 79.4% efficacy, assuming two rates of decline previously determined for tebuconazole (high decline rate), showed that the programme may remain profitable during the first 5 to 7 years of use, in contrast to cyproconazole (low decline rate), a fungicide that would be profitable during the entire decade. This approach was shown to be useful and can be adapted to other diseases of soybean and other crops, as long as damage functions are available. An interactive web app was developed to perform the simulations accessible at alvesks.shinyapps.io/rusty-profits/.     

Measuring lesion attributes and analysing their spatial patterns at the leaf scale using digital image analysis

Digital image analysis was used to quantify size, shape and relative positions of individual plant disease lesions to determine their spatial distribution pattern at the leaf scale. Rice brown spot was used as a necrotrophic pathogen causing numerous discrete lesions. A 50‐leaf subsample was selected from an existing data set of 350 images of leaves taken from the field, and analysed for disease severity using image analysis. Further measurements included size, shape and the relative positions of lesions for all leaves with severity > 8% (n = 25) and an additional 25‐leaf sample with severity <8%. A total of 3964 necrotic and/or halo areas were selected using a manually defined threshold in the computer program Assess . There were significant and positive associations (Pearson's r > 0.81; P < 0.001) between the size‐related measurements (lesion area, longest and shortest axis). Coalesced areas, formed by interconnection of lesions and associated haloes, and a high number of small lesions were found with an increase in severity, suggesting a secondary cycle and autoinfection process. Results from quadrat‐based (Poisson distribution and Spatial Analysis by Distance IndicEs) and distance‐based (point‐process Poisson) spatial methods were in good agreement and, together with a Taylor power law model, suggested a shift from random to predominantly aggregated patterns of lesions at severities approaching 10%. This framework, which is applicable to other foliar diseases, proved useful in providing quantitative knowledge of epidemic processes at the leaf scale. Finally, these results may be useful in improving simulation models and disease assessment methods.     

Development and evaluation of a standard area diagram set for the severity of phomopsis leaf blight on eggplant

A standard area diagram set (SADs) with eight severity values ranging from 0.5 to 32% was evaluated as a tool to improve the accuracy of the estimates of Phomopsis leaf blight severity on eggplant by ten inexperienced raters. A first assessment in a 50-leaf dataset of digital images was performed unaided (UN, no SAD). Two further aided (A) assessments, conducted two (A1) and four weeks after the first (A2) unaided one were performed. The precision of the estimates, as indicated by the Pearson’s correlation coefficient (r), improved significantly when using the aid (A1: r = 0.96 and A2: r = 0.96) compared to unaided (UN: r = 0.79). However, the generalized bias (C _b) was not significantly affected because the unaided estimates were already quite accurate (C _b = 0.93). The overall concordance (ρ _c) was significantly improved due to the large gains in precision when using the SADs. The raters’ estimates were more uniform (ρ _c > 0.9) when using the aid, with progressive gains in the reliability of the estimates among them, indicated by the concordance correlation coefficient statistics. The SADs proposed in this study will be useful in severity estimation during field work involving multiple raters, especially as use of the SADs results in less variable estimates, improving accuracy and reliability among raters.