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Resonant laser ionization and spectroscopy are widely used techniques at radioactive ion beam facilities to produce pure beams of exotic nuclei and measure the shape, size, spin and electromagnetic multipole moments of these nuclei. However, in such measurements it is difficult to combine a high efficiency with a high spectral resolution. Here we demonstrate the on-line application of atomic laser ionization spectroscopy in a supersonic gas jet, a technique suited for high-precision studies of the ground- and isomeric-state properties of nuclei located at the extremes of stability. The technique is characterized in a measurement on actinium isotopes around the N=126 neutron shell closure. A significant improvement in the spectral resolution by more than one order of magnitude is achieved in these experiments without loss in efficiency.

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In April 2010, pink-orange spore masses that later turned brown were observed on 7 to 50% of the transplant lots during a routine screening of Belgian strawberry (Fragaria × ananassa, cv. Elsanta) for the latent presence of Colletotrichum acutatum using the petiole freeze method (4). These spore masses contained hyaline, canoe-shaped to allantoid conidia (mean size 7.5 × 1.8 µm), which is not consistent with C. acutatum spore morphology. Subsequently, a spore mass was transferred onto potato dextrose agar (PDA) and a gray-to-brown colony with whitish, aerial mycelium was produced, which is also not consistent with C. acutatum isolates. To identify the fungus, the ITS1-5.8S-ITS2 rDNA region was amplified by PCR and sequenced. The 485-bp region was 100% identical to that of Pilidium concavum specimen voucher BPI 1107275 (GenBank Accession No. AY487094). P. concavum (Desm.) Höhn. (synanamorph Hainesia lythri; teleomorph Discohainesia oenotherae) is a pathogen of strawberry causing tan-brown rot of fruit and is a common secondary invader of roots and dead strawberry plant parts (3). A recent strain of P. concavum from strawberry, isolate UPL 50, obtained from Brazil (L. Zambolim, Univ. Fed. de Viçosa, personal communication) showed similar colony, microscopic (mean spore size of 6.8 × 1.8 µm), and molecular (ITS sequence 98% identical to that of P. concavum specimen voucher BPI 1107275) features as the Belgian isolate. Pathogenicity tests were conducted on mature strawberry fruits by submerging 15 fruits per isolate for 3 min in a conidial suspension (2 × 106 conidia ml-1 of water) obtained from a 2-week-old colony on PDA. Controls were submerged in sterile distilled water. The inoculated fruits were incubated in a moist chamber at 25°C. Sunken, yellowish brown lesions with pink and later orange-brown spore masses were observed starting 3 days after inoculation on 88 and 94% of the fruit for the Brazilian and Belgian isolate, respectively. The control fruits remained healthy. The fungal isolates were reisolated from symptomatic fruits and their identity was confirmed based on morphological features. During a strawberry field survey in July 2010 in Sint-Truiden (Belgium), lesions typical of those described above were observed on eight strawberry fruits (cv. Elsanta). The fungus was isolated from the symptomatic tissue of two fruits and characterized as described above. Since P. concavum was latently present on strawberry transplants and caused disease on the fruits in the field, we conclude that P. concavum is a potential threat for Belgian strawberry production. Moreover, no strawberry cultivars with resistance to the pathogen have been reported. The disease has previously been reported on strawberry in South America and Poland (1,2), but to our knowledge, this is the first report of P. concavum on strawberry in Belgium. Although the spore and colony morphology of P. concavum is different from C. acutatum, the spore masses of P. concavum can easily be confused with the spore masses of C. acutatum when using the freeze method. This suggests the need for microscopic analysis of these spore masses during routine analyses. References: (1) L. Cedeno et al. Interciencia 26:113, 2001. (2) U. P. Lopes et al. New Dis. Rep. 21:7, 2010. (3) J. L. Maas. Compendium of Strawberry Diseases. The American Phytopathological Society St. Paul, MN, 1998. (4) J. C. Mertely and D. E. Legard. Plant Dis. 88:407, 2004.

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Estimates of ascospore maturity generated by models developed for Venturia pyrina in Victoria, Australia (NV and SV), Oregon, United States (OR), and Italy (IT) or for V. inaequalis in New Hampshire, United States (NH-1) or modified in Norway (NH-2) were compared with observed field ascospore release of V. pyrina from 21 site-year combinations. The models were also compared with ascospore release data from laboratory assays. In the laboratory assays, the forecasts of the NH-1 and NH-2 models provided the best fit to observed spore release. Under field conditions, the lag phases and slope coefficients of all models differed from those of observed release of ascospores. Identifying the precise time of bud break of pear to initiate degree-day accumulation was problematic at both Australian sites. This resulted in a higher deviance between bud break and first released ascospore compared with the sites in Norway and Belgium. Linear regressions of observed release against forecasted maturity generated similarly high concordance correlation coefficients. However, where differences were noted, they most often favored models that included adjustment for dry periods. The NH-2, IT, and NV models using pooled data also provided the most accurate estimates of 95% ascospore depletion, a key event in many disease management programs.

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Scab is one of the key parasites in fruit growth. In favourable weather conditions for the pathogen, a complete harvest can be destroyed if no control measurements are undertaken. The scab fungi on pear and apple are two distinct species. They have, however, a similar biological cycle. Despite the similarities, there are also clear differences and these differences are significant for the control of the pathogen. Pear scab does not only infect fruit and leaves as apple scab does, but also infects twigs. Especially in organic fruit growing, twig scab is a big problem. Once twig scab occurs, it seems to be impossible to get rid of scab in these orchards. The only possibility for the fruit grower in this case is a strict spraying schedule to ensure no further spread of the infection. The main goal of the project is a thorough study of the pear scab fungi (biology, sensitivity of different plant parts and cultivars, dispersal of the fungi and infection conditions, the pathogenicity and characterization of different biotypes) to unravel the life of the fungi and to develop a better control strategy. A better control strategy means a reduced fungicide use and a reduction of fungicide residue on the fruits at harvest, without a reduction of the quality of the fruits and cost effectiveness for the fruit grower. Special attention in the project goes to the role and the control of twig scab. The first results of this project will be shown.

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In fruit growing preharvest sprayings in the orchard are mainly applied to protect fruit from decaying. Next to multisite fungicides (captan, thiram, tolylfluanid) the most commonly used products recognized for the Belgium market are Bellis (pyraclostrobin & boscalid) and the combination of Topsin M (thiophanate-methyl) and Frugico (diethofencarb). In general the spraying schedule varies depending on weather conditions (infection risk), preharvest interval of available fungicides, fruitgrower and cultivar of pome fruit (apple/pear). Facing the climatological conditions before picking the residue loading on the fruit surface can differ enormously. Also wet (pre)grading is considered to decrease the product residue resulting to fruits which are less protected before entering the cold storage room. In this context a partially replacement of the preharvest treatments by one postharvest application could offer a reliable alternative to the PPP reduction program (Plant Protection Products) in the orchard. A standardized application method by dipping or drenching will cover the fruits homogenically resulting in a rationalized fungicide use compared to the preharvest sprayings in the orchard. For the Belgium market Philabuster (imazalil & pyrimethanil) is registered for postharvest treatments since for this product a proper solution for the waste water of postharvest uses was developed to protect surface waters (Funds technology). Philabuster provides an advanced mould control towards fruit rot pathogens Gloeosporium spp., Botrytis cinerea and Penicillium spp. In this context several trials were set up to evaluate the biological efficacy of Philabuster alone or in combination with preharvest sprayings in the orchard. In concrete different preharvest spraying schedules were applied in the last six weeks before harvest on apple and pear facing parameters as rational fungicide use, antifungal effectiveness and cost price. The purpose was to select the optimal combination in use of preharvest fungicides with Philabuster as postharvest treatment, which offer full protection towards all key pathogens in apple and pear.

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As new fungicides are mainly unisite action fungicides, the problem of fungicide resistance development is becoming more important every year. Combining chemical fungicides, which is the best anti-resistance strategy, is not always possible or recommended in the case when the number of available chemical fungicides are limited or a reduction in fungicide use is asked for. Therefore the use of salts as an anti-resistance strategy was looked upon. The salts evaluated were K(HCO3), KH2PO3, KHPO4 and K2SiO3. When using these salts as an anti-resistance strategy the efficacy obtained when spraying the compounds alone was often to low to be used in rotation with chemical fungicides. Only with K(HCO3)2 a good efficacy can be observed in some years. The variation in efficacy with K(HCO3)2 observed is higher for powdery mildew. Chitosan was also included in the trials against powdery mildew, however chitosan had no effect on the infestation.

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Stemphylium conidia and Pleospora ascospores were monitored in pear orchards in the region of Haspengouw in Belgium during the infectious periods of brown spot disease (end May - end August) in the years 2002, 2004 and 2005. The seasonal and daily dynamics of the captured fungal spores are discussed and a correlation analysis was performed to determine possible correlation with weather parameters. Furthermore the fungicide cover obtained by spraying upon climatological infection risk is compared with the inoculum pressure in the orchards. Pleospora ascospores were found until the end of May and discharge took place during rain events at the same time-points when Venturia inaequalis ascospores were detected. Although the fungi ejected during the same time period, the relative importance of the different spore peaks differed. The first conidia were detected at the end of May. After that date conidia were found almost every day. Seasonal dynamics of the conidia clearly differed between the years and also the number of spores retrieved differed. No spores were found below 7.5 degrees C and only at a temperature above 12.5 degrees C an increase in conidia was observed. When looking at the daily dynamics, a significant negative correlation was found between the aerial spore concentration and relative humidity and leaf wetness, and a significant positive correlation with wind, temperature and water vapor pressure deficit. When spraying upon a BSP-cast CR threshold value of 0.4, all days with spore discharges above 3 conidia/m3/day were covered in 2002 and 2004. In 2005, a year with very low infection risk and infestation, two periods with high spore discharge were not covered. The observations made show that incorporation of the inoculum pressure into the infection models will probably not lead to a big improvement of the infection models.

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Scab (Venturia inaequalis) is the principal disease endangering both integrated and organic apple production. Scab pressure tends to build up over the years and organic farmers rely mainly on copper and sulphur treatments for control. The use of Cu in crop protection received scrutiny in recent years as this metal tends to accumulate in soil and substrates. A number of alternative organic control substances have been proposed, with variable success in scab control. We investigated the effect of these alternative organic scab control measures on several apple varieties with low scab susceptibility. The choice of scab treatments had important effects on the mineral composition of leaves and fruits. As these values affect current and future yield in perennial crops, as well as storage quality, the use of certain scab control agents requires corrective application of nutrients during and in-between growth seasons.

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PHILABUSTER is a new post-harvest fungicide developed by Janssen Pharmaceutica N.V.. It provides an advanced mould control by post-harvest treatments of citrus and pome fruit. The product is formulated as a stable suspension concentrate intended for dilution in water before use. PHILABUSTER 400 SC contains 200 g/L imazalil and 200 g/L pyrimethanil. Both active ingredients have a different single site mode of action. Imazalil inhibits ergosterol biosynthesis (DMI), whereas pyrimethanil interferes with fungal enzyme secretion and methionine biosynthesis. Due to the combination of these low risk fungicides a good anti-resistance management can be obtained. In case of existing reduced sensitivity of a population to DMI or MBC fungicides, no cross-resistance with pyrimethanil was observed. PHILABUSTER showed good activity by post-harvest treatment against key pathogens on apple and pear Penicillium expansum (blue mold), Botrytis cinerea (gray mold) and Gloeosporium spp. (lenticel rot) in small and large scale experiments with artificial or natural infections. By dip treatment of large volumes of fruit (up to 50 tons) the depletion of both active ingredients in the treatment water was low, both when plastic or wooden bins were used. Lower dose rates resulted in an inferior and inconsistent residue level of both active ingredients on fruit. Possible advantages of post-harvest treatments versus field treatments for the control of storage diseases are discussed.

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The fungal pathogen Botrytis cinerea is the causal agent of grey mould, the most important fungal fruit rot disease in strawberry in Europe. Currently disease control for grey mould is based on preventive spraying every five to seven days during flowering and harvest. Replacing preventive spraying with applications based on infection warnings can optimize performance and reduce the amount of sprays needed. Success of this approach will depend on the accuracy of the model used to predict disease outbreak. For this reason three infection models (BOTEM, BoWaS, DSS-Italy) were evaluated during the growth seasons of 2003 and 2004. The experiments included June bearing, retarded June bearing and ever bearing strawberries. In all experiments the use of infection models leaded to a reduced number of fungicide applications. However the efficacy of the different models towards the control of B. cinerea also decreased compared to the efficacy obtained with a standard 7 day schedule. Best results were obtained with BOTEM, developed by HRI (Horticultural Research International, East-Malling, UK): 17-60% reduction in fungicide use and an efficacy between 66-93 depending on the growth season, culture practice and the fungicides used. Compared with routine preventive spraying, the Botrytis fruit rot percentage is slightly higher. A higher efficacy with Botrytis infection warnings can only be obtained if infection warnings change from curative to preventive. A retroactive evaluation of a preventive warning system was included. Making use of the 48h weather forecasts supplied by the Royal Meteorological Institute of Belgium (KMI) based on ALADIN for the region of Haspengouw, it was possible to replace 30 up to 100% of the curative application by preventive spraying depending on the experiment and the threshold set for the preventive model.

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Signum, a new fungicide developed by BASF, was applied during 6 successive years against fungal diseases in strawberries. The product is formulated as a water dispersible granule, containing 6.7 % pyraclostrobin and 26.7 % boscalid. Pyraclostrobin is similar in chemistry to other strobilurin fungicides like kresoxim-methyl and trifloxystrobin, registered for fruit disease control. Boscalid belongs to the class of carboxyanilides. Both components in the premix formulation combine two different biochemical modes of action in the fungal cell respiration. Therefore, this co-formulation gives a broad-spectrum activity and also a reduced resistance risk for different target pathogens. Botrytis cinerea is the most important disease on strawberry-fruits and thus the control of fruit rot is mainly focused on this fungus. In average over 6 years, Signum has not only given a very good control against Botrytis fruit rot, but it has also shown a high performance in the control of Colletotrichum. Besides, Signum provides good control of powdery mildew (Podosphaera aphanis) and limits the shift to other fruit rots like leather rot (Phytophthora cactorum and leak (Rhizopus, Mucor). The availability of several categories of fungicide families with a different mode of action gives opportunities in alternating different fungicides and is the best guarantee for a sustainable control of fruit rot in all kinds of strawberry production methods. Signum should be integrated in an overall disease management program. Trials, in which the applications of Signum were timed on disease forecasting, based on environmental factors favorable for Botrytis development, were very promising. This tool can also help in establishing the IPM-concept in the production of strawberries.

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We determined the human leucocyte antigen (HLA)-A, -B and -DR allele frequencies in recipients and donors of 115 cornea transplants, for recipients who developed graft rejection and those who did not. No difference in HLA allele frequencies of the recipients was found. The frequencies of the HLA-A26, -B35 and -B44 alleles in cornea donors were increased in recipients who developed graft failure. The detrimental effect on corneal graft survival of these alleles was significant (p < 0.001). No such effect was observed in renal transplantation. Corneal graft survival was similar when one or two A26, B35 or B44 alleles were present on the donor cornea. The negative effect was similar in magnitude to the previously reported negative effect of an HLA-B locus match between donor and recipient. When both a B-locus match and an A26, B35 or B44 allele were present, the negative effect on graft survival was twice as strong, indicating that different immune mechanisms are responsible for these phenomena.

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We determined HLA-A, -B, and -DR allele frequencies in kidney transplant recipients in relation to graft survival. Most recipients and donors were from African descent. The frequency of HLA-A30 was somewhat increased in recipients who rejected the graft. The frequency of HLA-B42 was significantly (P=0.002) increased in recipients who rejected the graft. Similar results were found for the HLA-DR3 allele; however, this effect was diminished when B42-, DR3+ individuals were analyzed. We further investigated the effect on transplant outcome of a unique African haplotype A30, B42, DR3, and of segments thereof, which have a high frequency in the local population. Log-rank analysis revealed that the negative effect on transplant outcome was least in A30-, B42+ recipients (P=0.417) and most pronounced in A30+, B42+ patients (P=0.006). We postulate that the negative effect on transplant outcome may reside in the A30, B42 segment of chromosome 6 and may be caused by a stronger than average immunoregulatory gene.

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Mixed lymphocyte reaction (MLR) assays were performed serially over 24 months in 19 first cadaver renal transplant recipients. Immunosuppression consisted of cyclosporine, methylprednisolone and azathioprine. Cyclosporine was withdrawn at 6 months postoperatively. The MLR reactivity gradually decreased over the first 3 months following transplantation. However, there was a significant increase in MLR reactivity at 12 months postoperatively after the cyclosporine withdrawal. This rebound effect in MLR reactivity following cyclosporine withdrawal could account for the increased incidence of acute rejection episodes.

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We analyzed the influence on allograft survival of pretransplant panel reactive antibodies (PRA) < 10%, PRA > 10%, autoantibodies, cold antibodies and a positive B cell crossmatch in 807 renal and 237 cardiac transplant recipients. Donors and recipients were predominantly of mixed ancestry (Khoi, San, Xhosa and Caucasoid). Log rank analysis showed that PRA < 10%, cold antibodies, and a positive B cell cross-match did not influence allograft survival. Autoantibodies were present only in renal recipients; they appeared to have a beneficial effect on allograft survival (P = 0.06). PRA > 10% appeared to have a detrimental effect on allograft survival in both renal (P = 0.07) and cardiac (P = 0.06) recipients. Since autoantibodies and PRA > 10% had opposing effects, the results of renal recipients were reanalyzed after omission of the recipients with autoantibodies and coexisting PRA > 10%. This resulted in augmentation of the protective effect for autoantibodies (P = 0.027) and of the detrimental effect for PRA > 10% (P = 0.020). Two-year survival curves showed that when autoantibodies coexisted with PRA > 10%, the long term, but not the short-term, detrimental effect of PRA > 10% was attenuated. Patients with a positive B cell cross-match clustered in the PRA > 10% group in both renal (PRA negative vs. PRA < 10%; P = 0.0251; PRA < 10% vs. PRA > 10%: P = 0.0011) and cardiac (PRA negative vs. PRA > 10%: P = 0.0085) recipients. We conclude that PRA > 10% is the best indicator to identify recipients at high risk for rejection, and that the influence of antibodies on graft survival can not reliably be established without taking coexisting antibodies into account.

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Serial mixed lymphocyte culture (MLC) was used to monitor the evolution of donor-specific responsiveness over the first 2 years after cadaveric renal transplantation. Lymphocytes obtained from 37 patients at 0, 1, 3, 6, 12, 18 and 24 months following transplantation were assayed in a one-way MLC using donor lymphocytes as stimulator cells. Donor-specific hyporesponsiveness developed in 66% of the patients with functioning grafts. Donor-specific sensitization was noted in 2 patients with functioning grafts and the MLC reactivity remained unchanged compared to the pretransplant value in 8 patients. The patients with donor-specific hyporeactivity tended to remain either free of rejection episodes or experienced early rejection episodes only. Thus, using serial MLC we were able to identify patients who developed hyporeactivity or tolerance to donor antigens. These patients may require less immunosuppression.

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The results of tissue typing on 115 recipient/donor pairs prior to corneal grafting were analyzed with the proportional hazard regression model for the incidence of the first rejection episode and for graft failure from rejection. Like other investigators, we found that a previously failed corneal graft and the degree of recipient corneal vascularization were significant risk factor for graft rejection. ABO blood group matching had no effect. The absence of mismatches in both the HLA-A and HLA-DR loci decreased the incidence of rejection. However, no difference was observed for the presence of one versus two mismatches. Regression results for the HLA-A and DR loci were not significant. Surprisingly, matching for one or both HLA-B alleles resulted in a significantly higher incidence of graft rejection episodes (P < 0.005) and of graft failure (P < 0.052). This adverse matching effect for the B locus was proportional to the number of mismatches.

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We studied C4A and C4B polymorphisms and HLA-B and -DR associations in the San, Khoi and Xhosa. C4A and C4B alleles were determined using conventional protein allotyping methods. The C4A*3, C4B*1 haplotype had a high frequency (30-55%) in all populations. The frequency of C4A*3, C4B*Q0 was 7-19%. The C4A*Q0, C4B*1 haplotype was frequent (15%) in the Khoi but very rare in the San (P < 0.001). C4A*12 A*91, C4B*Q0 was frequent in the Xhosa (15%) but rare in the San and Khoi (P < 0.001). Alleles C4A*5 and C4A*6, and the C4B*2 B*92 duplication were only found in the Xhosa. C4A alleles A*4, A*45, A*58, A*12, A*14, A*19 and the C4A*3 A*91 duplication were only found in the San/Khoi population group. In the San, fourteen extended haplotypes were found in a relatively high frequency (2-7%). In the Xhosa, one extended haplotype (B42, C4A*12 A*91, C4B*Q0, DR18) was found in a very high frequency (13%) and was characteristic for this group; five other extended haplotypes were found with a low frequency (< 3%).

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The genetic polymorphism of vitamin D binding protein (DBP) and of properdin Factor B (BF) was determined in unrelated Namibian San and Khoi, and in South African Blacks, Caucasoids and Cape Coloureds. Alleles have been confirmed by segregation patterns in family studies. The DBP phenotypes were identified by isoelectric focusing on ultrathin polyacrylamide gels and the BF phenotypes were identified by electrophoresis on 1% agarose gels; both methods were followed by immunofixation. The DBP and BF allele frequencies for all population groups were found to be in accordance with Hardy-Weinberg equilibrium. DBP*1S and BF*S allele frequencies in the San, Khoi and Blacks were similar; their frequency was far lower than in Caucasoids. The frequencies of the DBP*1F and BF*F were also similar in the San, Khoi and Blacks; however, the allele frequency was much higher in these groups than in Caucasoids. These differences were statistically significant (P < 0.001).

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