Ecological interactions,management lessons and design tools in tropical agroforestry systems

During the 1980s, land- and labor-intensive simultaneous agroforestry systems (SAFS) were promoted in the tropics, based on the optimism on tree-crop niche differentiation and its potential for designing tree-crop mixtures using high tree-densities. In the 1990s it became clearer that although trees would yield crucial products and facilitate simultaneous growing of crops, they would also exert strong competitive effects on crops. In the meanwhile, a number of instruments for measuring the use of growth resources, exploratory and predictive models, and production assessment tools were developed to aid in understanding the opportunities and biophysical limits of SAFS. Following a review of the basic concepts of interspecific competition and facilitation between plants in general, this chapter synthesizes positive and negative effects of trees on crops, and discusses how these effects interact under different environmental resource conditions and how this imposes tradeoffs, biophysical limitations and management requirements in SAFS. The scope and limits of some of the research methods and tools, such as analytical and simulation models, that are available for assessing and predicting to a certain extent the productive outcome of SAFS are also discussed. The review brings out clearly the need for looking beyond yield performance in order to secure long-term management of farms and landscapes, by considering the environmental impacts and functions of SAFS.

     

Effects of various densities of Ophiostoma ips inoculations on Pinus sylvestris in north‐western Spain

The aim was to determine the inoculation density above which Scots pine (Pinus sylvestris) is overcome by the blue‐stain fungus Ophiostoma ips that is associated with the bark beetle Ips sexdentatus. In north‐western Spain, stems of 16 Scots pines were inoculated at various densities (0, 400, 800 or 1600 inoculi/m²) along circumferential 100 or 150 cm wide inoculation belts. Each inoculum consisted of a 5 mm diameter cylinder of malt extract agar colonized by the fungus. Three months later, all trees were harvested and trunk resinosis and foliage colour were visually assessed. The percentage of healthy, desiccated, resin soaked, and blue‐stained sapwood, as well as growth productivity indices, were calculated from stem disks cut from within the inoculated zone of each tree. Sapwood‐specific hydraulic conductivity (Ks) of each tree was measured in the middle of the inoculated zone. All parameters of tree vigour changed dramatically to the worse when inoculation densities were above 400 inoculi/m², and foliage changed from green to yellow‐green or yellow when an inoculation density of 800 instead of 400 was used. The percentage loss of sapwood‐specific conductivity (PLC) increased from 30 to 90% and the percentage of healthy, conductive sapwood dropped from 85 to 35% at 800 inoculi/m². No effect of the width of the inoculation belt was observed, and there was no relationship between tree productivity indices and the level of resistance. A non‐linear negative relationship was found between PLC and the percentage of healthy sapwood. It is concluded that tree resistance was overcome and that trees were going to die when the inoculation density was ≥800 inoculi/m².     

The role of head computed tomography in equine practice

This retrospective study describes the computed tomography (CT) findings in 59 horses presented with diseases of the head over 8 years that underwent CT examination of this region, including dental or sinonasal diseases (Group A) (n = 42), osseous and/or articular diseases (Group B) (n = 11) and soft tissue diseases (Group C) (n = 6). For Group A, radiographic and CT findings comparison was possible. Computed tomography had higher sensitivity (100%) and specificity (96.7%) than radiography in diagnosing dental disease. Compared to CT, radiographic identification of sinus involvement was less sensitive, particularly for ventral conchal and sphenopalatine sinuses and presented an overall sensitivity of 43.5 and 16.7%, respectively. In Group B CT allowed identification of a higher number of bone fragments and fractures in the maxillary, lacrimal, sphenoidal, temporal and zygomatic bones not identified radiographically. Accurate identification of CT changes in the temporomandibular joint and temporohyoid articulation was also possible. Group C included both intra‐ and extra‐cranial disease, retrobulbar masses being the most representative pathology (n = 3). In this group, CT was considered the gold standard for detection of periorbital diseases. We conclude that CT is an imaging technique with high diagnostic value for evaluating the equine head, yielding additional information over multiple radiographic views, which may alter the outcome of the case. Additionally, this paper reports several conditions not previously described using CT.     

Lack of a type-2 glycosyltransferase in the fish pathogen Flavobacterium psychrophilum determines pleiotropic changes and loss of virulence

Flavobacterium psychrophilum is an important fish pathogen, responsible for Cold Water Disease, with a significant economic impact on salmonid farms worldwide. In spite of this, little is known about the bacterial physiology and pathogenesis mechanisms, maybe because it is difficult to manipulate, being considered a fastidious microorganism. Mutants obtained using a Tn4351 transposon were screened in order to identify those with alteration in colony morphology, colony spreading and extracellular proteolytic activity, amongst other phenotypes. A F. psychrophilum mutant lacking gliding motility showed interruption of the FP1638 locus that encodes a putative type-2 glycosyltransferase (from here on referred to as fpgA gene, Flavobacterium psychrophilum glycosyltransferase). Additionally, the mutant also showed a decrease in the extracellular proteolytic activity as a consequence of down regulation in the fpgA mutant background of the fpp2-fpp1 operon promoter, responsible for the major extracellular proteolytic activity of the bacterium. The protein glycosylation profile of the parental strain showed the presence of a 22 kDa glycosylated protein which is lost in the mutant. Complementation with the fpgA gene led to the recovery of the wild-type phenotype. LD₅₀ experiments in the rainbow trout infection model show that the mutant was highly attenuated. The pleiotropic phenotype of the mutant demonstrated the importance of this glycosyltranferase in the physiology and virulence of the bacterium. Moreover, the fpgA mutant strain could be considered a good candidate for the design of an attenuated vaccine.     

Variation in pathogenicity among the three subspecies of Phytophthora alni on detached leaves,twigs and branches of Alnus glutinosa

Pathogenicity tests were carried out on leaves, twigs and branches of Alnus glutinosa using several isolates of Phytophthora alni ssp. alni, P. alni ssp. multiformis and P. alni ssp. uniformis in vitro. Healthy fresh leaves were collected from disease‐free areas and inoculated with mycelium on agar discs or by dipping in zoospore suspensions. In addition, twigs and branches were collected from both disease‐free and disease‐affected areas, inoculated with mycelium on agar discs and incubated at four temperatures (15, 20, 25, 30°C). All subspecies tested were pathogenic but with varied level of virulence. In inoculation tests on foliage, wounding was a key factor in causing infections: lesions on inoculated wounded leaves were larger than on non‐wounded leaves. In the twig and branch inoculation tests, no differences in virulence were observed among the P. alni subspecies in terms of sampling locations, but lesions differed in size according to incubation temperature, with the largest lesions occurring on tissues incubated at 25°C. The work is the first to report foliar necrosis caused by P. alni on A. glutinosa. P. alni ssp. uniformis was the least virulent of the subspecies in branch inoculations. These findings demonstrate that various tissues of A. glutinosa could act as sources of pathogen inoculum and may disseminate alder Phytophthora in natural ecosystems.     

Kinetic characterization of the oxidation of chlorogenic acid by polyphenol oxidase and peroxidase. Characteristics of the o-quinone

Chlorogenic acid is the major diphenol of many fruits, where it is oxidized enzymatically by polyphenol oxidase (PPO) or peroxidase (POD) to its o-quinone. In spectrophotometric studies of chlorogenic acid oxidation with a periodate ratio of [CGA]0/[IO4-]0 < 1 and [CGA]0/[IO4-]0 > 1, the o-quinone was characterized as follows: lambda(max) at 400 nm and epsilon = 2000 and 2200 M-1 cm-1 at pH 4.5 and 7.0, respectively. In studies of o-quinone generated by the oxidation of chlorogenic acid using a periodate at ratio of [CGA]0/[IO4-]0 > 1, a reaction with the remaining substrate was detected, showing rate constants of k = 2.73 +/- 0.17 M-1 s-1 and k' = 0.05 +/- 0.01 M-1 s-1 at the above pH values. A chronometric spectrophotometric method is proposed to kinetically characterize the action of the PPO or POD on the basis of measuring the time it takes for a given amount of ascorbic acid to be consumed in the reaction with the o-quinone. The kinetic constants of mushroom PPO and horseradish POD are determined.     

Pear distillates from pear juice concentrate: effect of lees in the aromatic composition

Pear juice obtained from pear concentrate was fermented at room temperature using Saccharomyces cerevisiae (BDX, ENOFERM, France) as the fermentation microorganism. During the fermentation process, total sugars were measured. High performance liquid chromatography analyses were used to monitor the fermentation process and to characterize the pear wine. The pear wine obtained was distilled with its lees using three different equipments: a glass alembic (a glass pot still coupled to a glass column), a copper alembic, and a glass alembic with the addition of 5 g/L of copper shavings to the pot still. The same distillations were repeated with the wine without its lees (separated by decanting). Several distillation fractions were collected, up to a total of 500 mL of distillate. Gas chromatography was used to identify and quantify the volatile compounds in each fraction, and the methanol and ethanol contents. Based on these results, the heart fraction was defined. ANOVA tests were performed on the heart fractions to determine quantitative differences between some volatile compounds depending on the equipment used and the presence or absence of the wine lees. From this series of ANOVA tests, it can be concluded that the concentrations of the compounds that are considered to have a negative effect on the quality of the distillates (methanol, ethyl acetate, furfural) decrease or do not change when they are distilled in the presence of lees and in the copper alembic. In addition, the concentrations of the positive compounds (ethyl decanoate and ethyl-2-trans-4-cis-decadienoate) increase in the presence of lees for all of the equipment tested. So, it can be assumed that the distillation of pear wine with its lees in copper alembic leads to a better quality product.     

Characterization of protein fractions from Bt-transgenic and non-transgenic maize varieties using perfusion and monolithic RP-HPLC. Maize differentiation by multivariate analysis

Protein fractions from transgenic Bt and non-transgenic maize varieties, extracted by the Osborne solvent fraction procedure, were characterized for the first time by perfusion and monolithic RP-HPLC in very short analysis times. Albumins and globulins from different transgenic Bt maizes as well as from their non-transgenic isogenic varieties were eluted in four peaks using perfusion RP-HPLC, whereas prolamins and glutelins were separated in seven peaks. Monolithic RP-HPLC enabled the separation of maize proteins in a large number of peaks showing 6 and 10 main peaks for albumins and globulins, respectively. Prolamins migrated at retention times higher than 5 min as seven peaks, whereas glutelins were separated in three main peaks appearing at retention times higher than 6.0 min. Moreover, chromatograms of the whole protein extracts showed 8 and 11 components for perfusion and monolithic RP-HPLC, respectively. A comparison of the chromatograms of the whole protein extracts relative to transgenic and non-transgenic varieties evidenced quantitative differences on the percentages of area, mainly for peaks 2 and 3 by perfusion RP-HPLC and for peaks 3 and 7 by monolithic RP-HPLC. A discriminant analysis based on these proteic profiles was carried out to classify and predict transgenic Bt maize lines, achieving 100% correct classification using perfusion RP-HPLC.