The Use of a GUS Transformant of Trichoderma Harzianum, Strain T3a, to Study Metabolic Activity in the Spermosphere and Rhizosphere Related to Biocontrol of Pythium Damping-off And Root Rot

The activity of Trichoderma harzianum in the spermosphere and rhizosphere of different plant species was studied by use of a beta-glucuronidase (GUS) transformant (strain T3a). Hereby, direct observation of micro-habitats supporting metabolic activity of T. harzianum is reported. Germination of conidia and mycelial growth were not supported by exudates from healthy roots of various ages. Instead, growth and activity of T. harzianum depended on access to dead organic substrates such as seed coats, decaying roots, and wounds, including those caused by infecting pathogens. A correlation between the GUS activity of T. harzianum and the biomass of Pythium ultimum in infected roots was established. On the basis of our observations, we suggest that the biocontrol ability of T. harzianum involves competition with the pathogen for substrates including the seed coat, and wounded or infected root tissue.     

Effects of Salinity and Soil–Drying on Radiation Use Efficiency,Water Productivity and Yield of Quinoa (Chenopodium quinoa Willd.)

Drought and salinity reduce crop productivity especially in arid and semi‐arid regions, and finding a crop which produces yield under these adverse conditions is therefore very important. Quinoa (Chenopodium quinoa Willd.) is such a crop. Hence, a study was conducted in field lysimeters to investigate the effect of salinity and soil–drying on radiation use efficiency, yield and water productivity of quinoa. Quinoa was exposed to five salinity levels (0, 10, 20, 30 and 40 dS m^?1) of irrigation water from flower initiation onwards. During the seed‐filling phase the five salinity levels were divided between two levels of irrigation, either full irrigation (FI; 95 % of field capacity) or non‐irrigated progressive drought (PD). The intercepted photosynthetically active radiation was hardly affected by salinity (8 % decrease at 40 dS m^?1) and did not differ significantly between FI and PD. Radiation use efficiency of dry matter was similar between salinity levels and between FI and PD. In line with this, no negative effect of severe salinity and soil–drying on total dry matter could be detected. Salinity levels between 20 and 40 dS m^?1 significantly reduced the seed yield by ca. 33 % compared with 0 dS m^?1 treatment owing to a 15–30 % reduction in seed number per m², whereas the seed yield of PD was 8 % less than FI. Consequently, nitrogen harvested in seed was decreased by salinity although the total N‐uptake was increased. Both salinity and drought increased the water productivity of dry matter. Increasing salinity from 20 to 40 dS m^?1 did not further decrease the seed number per m² and seed yield, which shows that quinoa (cv. Titicaca) acclimated to saline conditions when exposed to salinity levels between 20 and 40 dS m^?1.     

Actinobacillus pleuropneumoniae osteomyelitis in pigs demonstrated by fluorescent in situ hybridization.

Necrotizing osteomyelitis and fibrinopurulent arthritis with isolation of Actinobacillus pleuropneumoniae serotype 2 is reported in two pigs from a herd with lameness and mild coughing problems among 8 to 12-week-old pigs. Application of fluorescent in situ hybridization targeting 16S ribosomal RNA of A. pleuropneumoniae in formalin-fixed tissue was performed to verify the association of A. pleuropneumoniae with the bone and joint lesions. By in situ hybridization A. pleuropneumoniae was demonstrated as multiple microcolonies or single cells dispersed in focal fibrinonecrotizing pleuropneumonia, in joints with arthritis, and in bone necroses including lysis of growth plate and suppurative inflammation in the adjacent trabecular metaphysis, thus demonstrating that well-known infections manifest new, unusual lesions.     

The influence of diet on Lawsonia intracellularis colonization in pigs upon experimental challenge

The objective of this investigation was to study if different feeding strategies influence experimental infections of pigs with Lawsonia intracellularis, the causative agent of proliferative enteropathy. In three sequential trials, a total of 144 weaned pigs were fed five different diets all made from a standard diet based on wheat and barley as carbohydrate source and soybean as protein source. The five diets were: a standard diet (fine ground and pelleted), the standard diet fed as fermented liquid feed, the standard diet added 1.8% formic acid, the standard diet added 2.4% lactic acid and a diet similar to the standard diet (made from the same ingredients), but fed coarse ground. Twenty-four pigs on each diet were orally inoculated with L. intracellularis and growth performance and faecal excretion of bacteria were monitored. Twenty-four pigs fed the standard diet were included as not experimentally infected controls. Pigs in the first two trials were sacrificed 4 weeks post-inoculation, whereas animals in the third trial were sacrificed after 5 weeks. Pigs in all experimentally infected groups excreted L. intracellularis. The fermented liquid diet delayed the excretion of L. intracellularis and furthermore, pigs fed the standard diet supplemented with lactic acid had limited pathological lesions when the intestines were examined 4 weeks after inoculation. The growth performance was reduced in pigs experimentally challenged with L. intracellularis, however the prevalence and severity of diarrhea was limited.     

Deficit irrigation based on drought tolerance and root signalling in potatoes and tomatoes

Agriculture is a big consumer of fresh water in competition with other sectors of the society. Within the EU-project SAFIR new water-saving irrigation strategies were developed based on pot, semi-field and field experiments with potatoes (Solanum tuberosum L.), fresh tomatoes (Lycopersicon esculentum Mill.) and processing tomatoes as model plants. From the pot and semi-field experiments an ABA production model was developed for potatoes to optimize the ABA signalling; this was obtained by modelling the optimal level of soil drying for ABA production before re-irrigation in a crop growth model. The field irrigation guidelines were developed under temperate (Denmark), Mediterranean (Greece, Italy) and continental (Serbia, China) climatic conditions during summer. The field investigations on processing tomatoes were undertaken only in the Po valley (North Italy) on fine, textured soil. The investigations from several studies showed that gradual soil drying imposed by deficit irrigation (DI) or partial root zone drying irrigation (PRD) induced hydraulic and chemical signals from the root system resulting in partial stomatal closure, an increase in photosynthetic water use efficiency, and a slight reduction in top vegetative growth. Further PRD increased N-mineralization significantly beyond that from DI, causing a stay-green effect late in the growing season. In field potato and tomato experiments the water-saving irrigation strategies DI and PRD were able to save about 20-30% of the water used in fully irrigated plants. PRD increased marketable yield in potatoes significantly by 15% due to improved tuber size distribution. PRD increased antioxidant content significantly by approximately 10% in both potatoes and fresh tomatoes. Under a high temperature regime, full irrigation (FI) should be undertaken, as was clear from field observations in tomatoes. For tomatoes full irrigation should be undertaken for cooling effects when the night/day average temperature >26.5 °C or when air temperature >40 °C to avoid flower-dropping. The temperature threshold for potatoes is not clear. From three-year field drip irrigation experiments we found that under the establishment phase, both potatoes and tomatoes should be fully irrigated; however, during the later phases deficit irrigation might be applied as outlined below without causing significant yield reduction:

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Potatoes

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After the end of tuber initiation, DI or PRD is applied at 70% of FI. During the last 14 days of the growth period, DI or PRD is applied at 50% of FI.

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Fresh tomatoes

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From the moment the 1st truce is developed, DI is applied at 85-80% of FI for two weeks. In the middle period, DI or PRD is applied at 70% of FI. During the last 14 days of the growth period, DI or PRD is applied at 50% of FI.

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Processing tomatoes

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From transplanting to fruit setting at 4th-5th cluster, the PRD and DI threshold for re-irrigation is when the plant-available soil water content (ASWC) equals 0.7 (soil water potential, Ψ_soil = −90 kPa). During the late fruit development/ripening stage, 10% of red fruits, the threshold for re-irrigation for DI is when ASWC = 0.5 (Ψ_soil = −185 kPa) and for PRD when ASWC (dry side) = 0.4 (Ψ_{soil, dry side} = −270 kPa).

The findings during the SAFIR project might be used as a framework for implementing water-saving deficit irrigation under different local soil and climatic conditions.     

Selection of biological control agents for controlling soil and seed-borne diseases in the field

Different screening methods for selection of biological control agents (BCAs), for controlling soil and seed-borne diseases, are discussed. The shortcomings of laboratory methods focused on mechanism of action are discussed and we conclude that these methods should be used with caution if candidates with multifactorial or plant mediated mechanisms of control are to be obtained. In vitro screens may be useful for specific groups of microorganisms, thus, screens for antibiotics may be relevant for Streptomyces spp., and promising results have been obtained using soil plating or precolonized agar methods to screen for mycoparasitism and competitive saprophytic ability. Experience with screening in the Nordic programme Biological control of seed borne diseases in cereals is summarized. Research in the four participating countries – Finland, Sweden, Norway and Denmark – followed the same paradigm: that of obtaining antagonists, well adapted to different Nordic environments, and developing them as effective BCAs. Potential antagonists were isolated from different sources and in planta screening methods were developed in order to optimize selection of antagonists effective against a range of seed borne pathogens. Screens in the laboratory or greenhouse were followed by screening in the field. The different screening procedures are compared and evaluated.     

Fermented liquid feed and fermented grain to piglets- effect on gastrointestinal ecology and growth performance

A study was carried out to determine the microbial and nutritional characteristics of dry feed, liquid feed containing fermented liquid cereal grain, and fermented liquid compound feed, and their effect on gastrointestinal ecology and growth performance of piglets. Three dietary treatments were designed: dry feed (DRY); fermented liquid cereal grain feed (FLG); and fermented liquid feed (FLF). The FLF was prepared by removing three times daily 50% of the mixture (feed and water) stored in a tank at 20 °C and replacing it with an equal amount of fresh feed and water. The FLG was similarly prepared but fermenting only the cereals (barley and wheat) and adding the remaining ingredients immediately before feeding. One hundred and twenty weaners were fed the diets during 6 weeks from weaning. The dietary content of lysine (g/100 g protein) was 6.01 in the DRY diet, 6.21 in the FLG diet, and 5.46 in the FLF diet. Feeding piglets with the FLG diet resulted in a higher density of yeasts, a higher concentration of ethanol in the GIT, and a change of the bacterial population of the stomach compared to feeding FLF. The FLG group had a numerically higher daily feed intake and daily body weight gain compared to the FLF group, but the values were not significant. Feeding liquid feed containing fermented liquid cereal grains as a means of avoiding microbial degradation of free amino acids in the feed and increasing feed intake by improving palatability seems promising.     

Evaluation of the urinary Cortisol: creatinine ratio in the diagnosis of hyperadrenocorticism in dogs

The diagnostic accuracy of the urinary cortisol:creatinine ratio (CCR), with the cortisol being measured by ELISA, was evaluated by subjecting data from 18 dogs with and 20 dogs without hyperadrenocorticism to recelver operating characteristic (ROC) curve analysis. The area under the ROC curve (W 0–93, SE_w 0–044) was much higher than 045, indicating that the CCR did distinguish between dogs with and without hyperadrenocorticism.A cutoff value of about 60 × 10^-6 was assoclated with the highest sensitivity (1.0)and speciflcity (0–85). At the disease prevalence rate of the present study (0 47), the positive and negative predictive values were 0–87 and 1.0, respectively. These numbers indicate that canine hyperadrenocorticism may be safely excluded when the CCR Is below 60 × 10^-6 but that a test of higher specificlty (eg, the ACTH stimulation test) should be used to confirm the diagnosis of canine hyperadrenocorticism when the CCR Is above 60 × 10^-6.