Hapten synthesis and monoclonal antibody-based immunoassay development for the detection of the fungicide kresoxim-methyl

Strobilurin fungicides have been increasingly used for fungus pest control since they were introduced in 1996. For pesticide residue detection, immunoassays constitute nowadays a valuable approach. This paper describes the synthesis of functionalized haptens of kresoxim-methyl, the production of monoclonal antibodies, and the development of enzyme-linked immunosorbent assays. On the one hand, a two-step conjugate-coated immunoassay was optimized using extended or short incubation times, with limits of detection of 0.4 ng/mL for the extended assay and 0.3 ng/mL for the rapid assay. On the other hand, an immunoassay was optimized following a procedure consisting of just one incubation step. This one-step assay had a limit of detection of 0.4 ng/mL. All of these assays showed a similar performance, with sensitivities well below common maximum residue limits for this pesticide (50 microg/kg) and lower than the detection limits of the usual chromatographic detection methods.     

Production and characterization of monoclonal antibodies specific to the strobilurin pesticide pyraclostrobin

Strobilurin fungicides are nowadays among the most important fungicides in the market of active agrochemicals. Pyraclostrobin, which belongs to the last generation of this family of molecules, shows a broader antifungal activity spectrum and higher efficiency and security profiles than previous fungicides. This paper describes the synthesis of functionalized haptens, the production of monoclonal antibodies, and the development of enzyme-linked immunosorbent assays (ELISA) for the detection of pyraclostrobin. A conformational analysis of hapten structure was performed, which provided relevant data concerning the length of the spacer arm. A very useful strategy has been followed for the screening of hybridomas, leading to the selection of a panel of high-affinity monoclonal antibodies to pyraclostrobin. Moreover, different immunoassays have been characterized using the conjugate-coated indirect ELISA format, and limits of detection below 0.1 microg/L have been obtained. Also, a simplified one-step procedure has been carried out with two indirect assays. Finally, these results have been compared with the performance of the same antibodies in the antibody-coated direct ELISA format.     

Carnation Fusarium wilt suppression in four composts

Fusarium wilt is now a major disease of carnation crops worldwide. Methyl bromide, which is used to remedy it, is environmentally unsafe. An alternative approach integrated into biological control is to grow crops in suppressive media. Suppressiveness of seven plant growth media to Fusarium oxysporum f. sp. dianthi was evaluated in bioassays with carnation (Dianthus cariophyllus) cv. Medea. These media were: (1) grape marc compost, (2) cork compost, (3) olive oil husk + cotton gin trash composted and mixed with rice husk, (4) spent mushroom compost mixed with peat, (5) coir fibre, (6) light peat and (7) vermiculite. In order to look for carnation Fusarium wilt suppressiveness indicators, growth medium pH and β-glucosidase activity were evaluated. Furthermore, F. oxysporum populations were measured in plant growth media at the beginning and end of bioassays. The compost media showed a range of suppressiveness in comparison with peat. Grape marc compost was the most effective plant growth medium in suppressing carnation Fusarium wilt. On the other hand coir fibre, peat and vermiculite were conducive for this disease. β-glucosidase activity and pH were positively correlated with disease severity as in other reports for tomato. Therefore, these two parameters are good indicators for carnation Fusarium wilt suppressiveness, and possibly for other F. oxysporum pathosystems. All composts showed similar F. oxysporum populations at the end of the bioassays to peat and vermiculite.     

Scintigraphic abnormalities of the pelvic region in horses examined because of lameness or poor performance: 128 cases (1993-2000)

OBJECTIVE: To identify scintigraphic abnormalities in the pelvic region of horses examined because of hind limb lameness or poor performance and determine the clinical relevance of areas of abnormal radiopharmaceutical uptake (ARU) in these horses. DESIGN: Retrospective study. ANIMALS: 128 horses. PROCEDURE: Medical records were reviewed, and information on signalment, history, admitting complaints, physical examination findings, and results of lameness examinations was recorded. Clinical relevance of areas of ARU was determined by comparison with results of other diagnostic tests. For horses with clinically relevant areas of ARU, follow-up information was obtained through telephone interviews with owners and trainers and analysis of race records. RESULTS: Areas of ARU were identified in the tuber coxae (25 horses), ischiatic tuber (9), hip joint (10), third trochanter (10), ilium (5), sacral tuber region (22), greater trochanter (1), cranial femoral cortex (1), skeletal muscle surrounding the pelvis (34), or multiple areas (11). In 44 horses, areas of ARU were associated with the primary cause of lameness; in 51, areas of ARU were not associated with the primary cause of lameness; and in 33, the primary cause of lameness was not determined. Thirty-six of the 44 horses with clinically relevant areas of ARU were available for follow-up; 15 (42%) had a good outcome. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that pelvic scintigraphy may be useful in identifying abnormalities in horses with hind limb lameness or poor performance.     

Predictive factors for the suppression of fusarium wilt of tomato in plant growth media

ABSTRACT Fusarium wilts are economically important diseases for which there are no effective chemical control measures. However, biological control and fertility management are becoming efficient alternatives for controlling this disease. Growth media formulated with composts that are able to suppress Fusarium wilt of tomato provide a control system that integrates both strategies. The aim of this study was to predict Fusarium wilt suppression of growth media using abiotic and biotic variables. Grape marc compost was the most effective medium used to suppress Fusarium wilt. Cork compost was intermediate, and light peat and expanded vermiculite were the most conducive growth media. The growth media evaluated were in a pH range of 6.26 to 7.97. Both composts had high beta-glucosidase activity. When pH and beta-glucosidase activity were taken into account as predictive variables, more than 91% of the variation in severity of Fusarium wilt was explained. This relationship illustrates the effect of nutrient availability and the degree of microbiostasis, two key factors in this pathosystem. Microbial populations involved in suppressiveness were cellulolytic and oligotrophic actinomycetes, fungi, and the ratios cellulolytic actinomycetes/cellulolytic bacteria, oligotrophic bacteria/copiotrophic bacteria, and oligotrophic actinomycetes/oligotrophic bacteria. Based on community level physiological profiles, different community structures were evident among growth media evaluated.     

Involvement of cytolytic and non-cytolytic T cells in the control of feline immunodeficiency virus infection

The appearance of non-cytolytic T cells that suppressed feline immunodeficiency virus (FIV) replication in vitro, and FIV-specific cytotoxic T cell (CTL) responses was compared in a group of seven, specific pathogen free (SPF) domestic cats following primary infection with the Glasgow(8) isolate of FIV (FIV(GL-8)). FIV proviral burdens were quantified in the blood and lymphoid tissues by real-time PCR. Non-cytolytic T cell suppression of FIV replication was measured by co-cultivating lymphoblasts prepared from the cats at different time-points during infection with FIV-infected MYA-1 cells in vitro. Non-cytolytic suppressor activity was detected as early as 1 week after infection, and was evident in all the lymphoid tissues examined. Further, this activity was present in subpopulations of T cells in the blood with normal (CD8(hi)) or reduced (CD8(lo)) expression of the CD8 molecule, and temporal modulations in non-cytolytic suppressor activity were unrelated to the circulating CD8(+) T cell numbers. Virus-specific CTL responses, measured by (51)Cr release assays, were not detected until 4 weeks after infection, with the emergence of FIV-specific effector CTLs in the blood. Throughout infection the response was predominantly directed towards FIV Gag-expressing target cells, and by 47 weeks after infection CTL responses had become localised in the lymph nodes and spleen. The results suggest that both non-cytolytic T cell suppression of FIV replication and FIV-specific CTL responses are important cellular immune mechanisms in the control of FIV replication in infected asymptomatic cats.     

An Echocardiographic Study of Atrial Fibrillation in Horses: Before and After Conversion to Sinus Rhythm

Two-dimensional and M-mode echocardiograms were recorded from 41 horses before they were successfully treated for atrial fibrillation. In addition, these examinations were performed in a subgroup of 20 horses after treatment, and the results were compared with pretreatment values. Atrial fibrillation in this group of horses was associated with a reduction of mean left ventricular fractional shortening (mean 31 %± 5.24%), and 22 of the 41 horses were below the reference range. The remaining mean M-mode variables were within the normal reference range, although 12 horses had increased left ventricular lumen dimensions in systole, and 8 horses had decreased left ventricular ejection times. Abnormal motion of the mitral valve was present in all horses and was characterized by the absence of A peaks, which were replaced by small diastolic undulations in 55% of the horses. In horses 1 to 20, after conversion to sinus rhythm, the mean fractional shortening increased (35.34%± 5.4%, P = .004), but there were no significant differences in heart rate or left ventricular lumen diameters in systole or diastole. These results suggest that ventricular function may be compromised by the presence of atrial fibrillation. However, this improved after correction of the arrhythmia.     

Variation in carbon concentration and wood density for five most commonly grown native tree species in central highlands of Ethiopia: The case of Chilimo dry Afromontane forest

ABSTRACT

Information regarding carbon concentration and wood density are lacking in Chilimo dry Afromontane forest.

The aim of this study was to estimate carbon concentration and wood density for Allophyllus abyssinicus, Olea europaea, Olinia rochetiana, Rhus glutinosa, and Scolopia theifolia. A total of 105, 30–50 mm thick wood discs were collected and oven dried at 102°C and 67°C to constant weight, chopped and finally grinded into 0.2 mm with a grinding mill. Carbon concentration was analyzed using the ash method, while wood density was estimated using the water displacement method. The highest carbon concentration (57.12%) was found for O. rochetiana, however, the lowest carbon concentration (56.43%) was found for A. abyssinicus. Stem parts had higher carbon concentration (56.98%) than branch (56.74%) and leave (54.53%) parts. O. europaea exhibited the highest wood density (0.67 g cm^?3) value than other species. However, the lowest wood density (0.42 g cm^?3) was exhibited for A. abyssinicus. Wood density was also showed a decreasing trend along with increases in stem height and maximum wood density (0.62 g cm^?3) was found under stump position, while, the minimum wood density (0.4 g cm^?3) was found under tree commercial height.     

Effects of light/darkness, thermal shocks and inhibitory components on germination of Pinus canariensis, Pinus halepensis and Pinus pinea

Knowledge of the germination characteristics of Pinus species can help in the understanding, prediction and management of the regeneration of pine forests. In the Canary Islands, several exotic pines (Pinus halepensis L. and Pinus pinea L.) were planted with Pinus canariensis Chr. Sm. Ex DC, the only native pine species, and there is now an interest in controlling these exotic species to restore the original forest. The main objective of the present study is to determine the germination response of P. canariensis, P. halepensis and P. pinea to different light regimes (darkness and light/darkness), thermal shocks and the presence of inhibitory substances from the leaves and litter of P. canariensis. P. halepensis seeds showed the highest viability and germination rate. Darkness accelerated P. halepensis germination, while exudates accelerated P. canariensis germination. Only treatments of 200 °C for 5 min and 300 °C for 5 min significantly decreased the germination of all three pine species. In the absence of strong differences in germination among species after treatments, the key for fire management or prescribed burning in this case may be the sprouting ability of P. canariensis. A possible management strategy to control the two obligate seeder exotic species could be to use one intense prescribed fire followed by a low-intensity prescribed fire after seed bank germination in the field, so in this way, only P. canariensis could resprout after fire and it would eliminate only the exotic pines in a mixed stand.