Polyclonal and monoclonal antibodies specific to the chrysanthemic acid moiety of pyrethroid insecticides

Two polyclonal and two monoclonal antibodies raised against chrysanthemic acid (CAA) were prepared for ELISA of pyrethroid insecticides with the common CAA moiety. The monoclonal antibody KCA226 showed the highest reactivity towards allethrin among the antibodies in C-ELISA, although KCA226 was more sensitive to higher concentrations of methanol than the polyclonal antibodies in ELISA. KCA226 was reacted linearly with allethrin at concentrations ranging from 1 to 10 μg litre^-1 in 0·5% methanol and from 300 to 10000 μg litre^-1 in 30% methanol. KCA226 reacted specifically with the pyrethroid insecticides with the CAA moiety but was much less reactive with CAA itself. These results suggest that C-ELISA based on KCA226 is useful for the assay of pyrethroid residues with the common CAA moiety. © 1998 Society of Chemical Industry     

Enzyme immunoassay for atrazine analysis in water and soil

Enzyme immunoassay (EIA) has been tested for the detection of atrazine in soil and water. EIA kits and atrazine-fortified samples were received from the International Atomic Energy Agency. Atrazine concentrations of about 0·01 μg litre^-1 could be detected and the central detection point was found at about 0·15 μg litre^-1 which is a reasonably sensitive region for atrazine. A validation study with spiked local water samples yielded acceptable results. No treatment was required for water samples. Extraction of atrazine from soil was done by simple shaking with methanol without any clean-up steps. Detection limits of 1×10^-2 μg litre^-1 for water and 5×10^-3 μg kg^-1 for soil were achieved. © 1998 SCI.     

A monoclonal antibody‐based ELISA for the analysis of the insecticide flucythrinate in environmental and crop samples

A competitive enzyme‐linked immunosorbent assay (ELISA) has been developed for the detection of the insecticide flucythrinate in environmental and food samples. Two types of haptens, the acid moiety that is the hydrolyzed product of flucythrinate, and the carboxylated propyl derivative of the alcohol moiety, were used to prepare monoclonal antibodies (MAbs). Five MAbs, which raised against the former hapten, were reactive with flucythrinate. Among them, MAb F1A27‐4 showed the highest activity toward flucythrinate, and did not cross‐react with other pyrethroids such as cycloprothrin, fenvalerate, fluvalinate, etofenprox and silafluofen. The assay conditions of indirect competitive ELISA with MAb F1A27‐4 were studied to optimize the detection of flucythrinate in environmental and food samples. Incubation at 4 °C in the assay buffer, pH 8, with 300 mM sodium chloride improved the sensitivity. The addition of rabbit serum albumin or rabbit antiserum and the presence of 50 ml litre^?1 of methanol reduced matrix effects of the samples. Under optimized conditions, the ELISA detected flucythrinate spiked in water, soil, and extracts of apple and tea samples down to 10 mg litre^?1, 0.2 mg litre^?1, 0.3 mg litre^?1 and 0.3 mg litre^?1, respectively. The mean recovery and CV ranged from 91% to 120% and from 5% to 12%, respectively. The ELISA results in apple samples correlated well with those from LC–MS analysis (r² = 0.99, n = 12). © 2001 Society of Chemical Industry     

Determination of residual 2,4,5-T in vegetables by high performance liquid chromatography with ultraviolet and fluorometric detection after derivatization with 2-(2,3-naphthalimino)ethyl trifluoromethanesulfonate (NE-OTf)

2,4,5-T was extracted with acetone at below pH 1·0 and the extract was concentrated. After adding 100 g litre^-1 sodium chloride solution to the residual solution, 2,4,5-T was extracted with ethyl acetate+hexane (20+80 by volume). The extract was evaporated to dryness and the residue was dissolved in acetonitrile. 18-crown-6, potassium fluoride and NE-OTf were added to the acetonitrile solution and then allowed to react at 50°C for 20 min. The product was injected to a HPLC with ultraviolet detection operated at 259 nm and fluorometric detection at 394 nm emission and 259 nm excitation. The determination limits of the 2,4,5-T derivative in the sample were 20 μg litre^-1 with UV detection and 10 μg litre^-1 with fluorometric detection. © 1997 SCI.     

Leaching of atrazine and hexazinone from Abies nordmanniana (Steven) spach plantations

The content of the herbicides atrazine and hexazinone was measured in drainage water from seven to ten-year-old plantations grown with Abies nordmanniana (Steven) Spach on two clayey soils in Denmark. The concentrations of atrazine varied between 0.06 and 7.79 μg litre^?1. The concentrations of hexazinone were different at the two locations, ranging from 0.07 to 2.09 μg litre^?1 at Bremersvold and from 3.47 to 42.66 μg litre^?1 at Koege. Metabolites of both herbicides were identified.     

Influence of different field sites on pesticide movement into subsurface drains

Pesticide movement to subsurface drains was monitored in two typical crop production areas in Germany. Field trials were conducted on two subsurfacedrained soils, a silt loam and a poorly structured sandy soil, under different climatic conditions. Over a period of one year, the drainflow was measured and the drain water was analysed for all applied herbicides. Different leaching behaviour was observed at the two field sites. Following autumn application of pendimethalin and isoproturon to the Soester Börde soil, maximum concentrations of about 62 μg litre^?1 for isoproturon and 0.7 μg litre^?1 for pendimethalin were observed in drainflow from this silt loam. The early occurrence of both herbicides in the drain water only two days after application is consistent with fast flow through macropores. In contrast, on the subsurfacedrained sandy soil in Brandenburg, isoproturon did not reach the drains until two months after autumn application and was found at maximum concentrations of only 1.4 μg litre^?1; pendimethalin was not detected in the drain water. Pesticide movement after spring application seemed to be of minor importance. At both locations, spring application led to low concentrations of pesticides in the drainflow (pendimethalin < 0.01 μ litre^?1; metolachlor ? 0.05 μ litre^?1; chloridazon ? 0.15 μ litre^?1; metamitron ? 0.02 μg litre^?1; terbuthylazine ? 1.4 μ litre^?1).     

Morphogenetic effects of diflubenzuron,an inhibitor of chitin deposition,on the coconut black-headed caterpillar (Nephantis serinopa myer)

Feeding the last instar larvae of Nephantis serinopa, the coconut black-headed caterpillar, on coconut leaves that had been dipped in diflubenzuron suspensions (0.2-4.0 g litre^?1) and dried, inhibited the moulting process and caused morphogenetic deformities of their pupae. The inhibition of moulting ranged from 27.5 to 75% The maximum effect was produced by suspensions of 4.0 g litre^?1 when only 5% of the larvae survived to become adults against 97.5% of untreated larvae.     

Reaction of saprophytic bacteria from potato peel extracts and plant pathogenic bacteria in ELISA with antisera to Erwinia chrysanthemi (serogroup O1Ha)

The specificity of two antisera raised to whole cells ofErwinia chrysanthemi (Ech), serogroup O₁H_a, was studied in double antibody sandwich (DAS-) ELISA with 100 strains of different plant pathogenic bacteria (PPB), including 39 Ech strains, and of one of these antisera with 900 saprophytic bacteria isolated from extracts of potato peelings of Dutch seed potatoes grown in several production areas.All tested European Ech strains from potato reacted positively while no reactions were observed with any of the other plant pathogenic bacterial species. Two saprophytes (A254 and A256), both identified as pectinolyticPseudomonas fluorescens species, cross-reacted strongly with polyclonal antibodies against Ech. Non-specific reactions were found in DAS-ELISA with 16 saprophytes. The detection limits for the individual saprophytes varied between c. 10⁵ and 10⁹ cells.ml^–1. The non-specific reactions were also found with monoclonal antibodies (mca 2A4) against a proteinase K resistent epitope of Ech and with antisera against other plant pathogens including an antiserum against potato virus Y^N. The non-specific reactions were observed in DAS-ELISA, but not in Ouchterlony double diffusion or immunofluorescence colonystaining, whereas A254 and A256 reacted in all tests, but only with antibodies against Ech. When in making dilution series potato peel extracts were used instead of phosphate buffered saline with 0.1% Tween 20, the 14 non-specifically reacting saprophytes only reacted at concentrations of 10⁹ cells.ml^–1 or higher. Only one of these 14 saprophytes was able to multiply on injured potato tuber tissue.In contrast to most saprophytic strains, the saprophytes A254 and A256 reacted strongly in ELISA in dilutions series made with potato peel extracts. A256 was able to grow on potato tuber tissue but only under low oxygen conditions; A254 did not grow at all on potato tissue.Defatted milk powder or bovine serum albumin added to the dilution buffer for the enzymeconjugated antibodies, drastically reduced the non-specific reactions, but not the reactions with A254 and A256.To reduce the cross-reaction with A254, an Ech antiserum was absorbed with A254. This resulted in a substantial drop in antibody reaction with the homologous antigen in Ouchterlony double diffusion.     

DPX-MP062—a potent compound for controlling the Egyptian cotton leafworm Spodoptera littoralis (Boisd.)

The effects of DPX-MP062 [methyl 7-chloro-2,3,4a,5-tetrahydro-2-[methoxycarbonyl(4-trifluoromethoxyphenyl)carbamoyl] indeno[1,2-e][1,3,4] oxadiazine-4a-carboxylate] a broad-spectrum insecticide with a novel mode of action, on the Egyptian cotton leafworm, Spodoptera littoralis, were studied in laboratory experiments. Egg hatch was affected by high concentrations (125 mg AI litre^-1) of DPX-MP062. Larvae that hatched from treated eggs were significantly affected at concentrations of 12·5 mg AI litre^-1 and greater. Larvae were fed castor bean leaves treated with DPX-MP062; 1st-instar larvae were the most susceptible development stage. Pupation and adult formation were determined in assays with 5th-instar larvae. There was strong suppression of adult formation; 65 and 91% at 0·5 and 0·75 mg AI litre^-1, respectively. Highly affected larvae died before pupation; slightly affected ones reached pupation 2–4 days later, were smaller than larvae in the untreated control, and were sometimes unable to develop into normal adults. Comparatively high concentrations (50 and 100 mg AI litre^-1) of the test compound were necessary to affect adults by ingestion, but no effects from contact application could be determined at a concentration of 100 mg AI litre^-1. © 1998 Society of Chemical Industry     

Deposition of aerially applied tebufenozide (RH5992) on balsam fir (Abies balsamea) and its control of spruce budworm (Choristoneura fumiferana [Clem.])

A field trial was conducted in 1994 to determine the foliar deposit of tebufenozide (RH5992), applied aerially, and its efficacy against spruce budworm, Choristoneura fumiferana (Clem.). A commercial 240 g litre^-1 formulation of the insecticide (Mimic 240LV) was mixed with water, dyed with a tracer dye (Rhodamine WT) and sprayed with a light fixed-wing aircraft. Six application strategies were tested. Five used 70 g AI ha^-1 in a spray volume of 1 or 2 litre^-1 ha^-1 with single or double applications; the sixth was an unsprayed control. Results show that the spectra of the spray applications were, with one exception, fairly uniform. Volume and number median diameters ranged from 100 to 130 μm and 27 to 72 μm, respectively. Mean number of drops cm^-2 on Kromekote cards were <2·0 for strategies where either 1 or 2 litre ha^-1 were sprayed. Nevertheless no one strategy produced droplet densities that were significantly different (P<0·05) from the other strategies. Tebufenozide recovered from foliage averaged 2·5 to 5·9 μg g foliage^-1 when 1 litre ha^-1 was sprayed and 5·8 to 6·8 μg g foliage^-1 after 2 litre ha^-1 were sprayed. When a single application was the strategy used, the mean number of droplets cm^-2 and μg tebufenozide g foliage^-1 ranged from 1·2 to 1·4 and 2·5 to 5·9, respectively. With double applications, the same response parameters ranged from 0·3 to 1·9 and 2·5 to 6·8, respectively. Budworm population reductions (%) and the number of larvae that survived tebufenozide treatments were significantly different (P<0·05) from the controls. After strategies that used 1 litre spray ha^-1, mean percentage population reductions ranged from 61·4 to 93·6 whereas populations were reduced by 85·6 to 98·3% when 2 litre ha^-1 were sprayed. After double applications the mean percentage population reductions ranged from 93·6 to 98·3, but single application strategies resulted in mean reductions of 61 to 86%. Mean population reductions in the controls were 61%. The mean number of larvae per branch that survived spray strategies of 1 litre ha^-1 ranged from 1·3 to 7·4, and from 0·4 to 1·3 when 2 litre ha^-1 was the spray volume. In the controls an average of 10·2 larvae survived. With one exception, mean percentage defoliation in the treated areas was also significantly less (P<0·05) than that in the control. Mean defoliation in trees sprayed at 1 litre spray ha^-1 ranged from 40 to 62·8% whereas those treated at 2 litre ha^-1 had mean defoliation levels from 31·5 to 62·8%. In contrast, average defoliation in the controls was 92·1%. When a single application was the spray strategy, mean defoliation ranged from 31·5 to 62·8%. These data imply that a double application of tebufenozide at 70 g in 2 litre ha^-1 was the most efficacious strategy. However, analyses of the data also show that the primary influence on deposits and defoliation was interactions between number of applications and spray. Nevertheless the two independent variables acted without significant interactions when influencing percentage reductions of spruce budworm populations. © 1998 SCI     

Communication to the Editor Protection of Wood against the House Longhorn Beetle Hylotrupes bajulus with Sodium Chloride and Potassium Chloride

The effectiveness of aqueous solutions of sodium chloride and potassium chloride as wood preservatives was tested against larvae of the house longhorn beetle Hylotrupes bajulus (L.). Immersion of pine (Pinus sylvestris L.) blocks for 24 h provided only a shallow protective layer. In a test series according to EN 46 a concentration of 200 g litre^-1 potassium chloride was required for complete larval mortality within 12 weeks; 200 g litre^-1 sodium chloride failed to achieve this. With vacuum impregnation in tests according to EN 47 a concentration of 100 g litre^-1 sodium chloride achieved complete mortality of larvae within four weeks. Vacuum impregnation of 100 g litre^-1 potassium chloride attained complete mortality only after 12 weeks. Ovipositing female beetles did not avoid wooden blocks treated with either salt. © 1997 SCI.     

Gas-liquid chromatographic determination of hydrazine in maleic hydrazide formulations and in samples stored at an elevated temperature

A method is described for the analysis of small amounts of hydrazine in maleic hydrazide formulations. Following derivative formation with pentafluorobenz-aldehyde, the pentafluorobenzaldehyde azine was extracted with hexane and determined by gas-liquid chromatography with electron-capture detection. Recoveries of 72-80% were obtained from samples fortified with 1 and 10 μg of hydrazine. The limit of detection was 0.05 mg kg^?1. Fourteen commercial formulations with maleic hydrazide concentrations ranging from 180-360 g litre^?1 were investigated. The hydrazine content of the maleic hydrazide used in these formulations ranged from less than 0.05 to 53 mg kg^?1. During the storage of two samples at 50°C for 10 weeks, the hydrazine contents increased from 2.2 to 124 and 0.4 to 54 mg litre^?1, respectively.     

Effects of some nonionic polyoxyethylene surfactants on uptake of ethirimol and diclobutrazol from suspension formulations applied to wheat leaves

The influence of a number of commercial nonionic polyoxyethylene surfactants on the foliar penetration and movement of two systemic fungicides, ethirimol and diclobutrazol, was studied in outdoor-grown wheat plants at different growth stages and post-treatment temperatures in two consecutive growing seasons. Both fungicides were applied as ca 0·2 μl droplets of aqueous suspension formulations containing 0·5 g litre^?1 of ¹⁴C-labelled active ingredient; surfactants were added to these suspensions at concentrations ranging from 0·2-10 g litre^?1. To achieve optimum uptake of each fungicide the use of surfactants with different physicochemical properties was required. For diclobutrazol, a lipophilic compound, uptake of radiolabel was best with surfactants of low mean molar ethylene oxide (E) content (5-6) but it was necessary to use concentrations of ca 5 g litre^?1 to attain this. The surfactant threshold concentration for uptake enhancement of radiolabel from ethirimol formulations (< 2 g litre^?1) was much lower than that for diclobutrazol but surfactants with E contents > 10 induced the greatest amount of uptake. For both fungicides, surfactants with an aliphatic alcohol hydrophobe were generally more efficient in promoting their uptake than those with a nonylphenol moiety. The sorbitan-based surfactant ‘Tween 20’ proved to be an effective adjuvant only for the ethirimol formulation; the uptake enhancing properties of the block copolymer ‘Synperonic PE/F68’ were weak. Uptake performance could not be related to the spreading properties of the respective formulations on the wheat leaf surface or to differences in solubilisation of the two fungicides by the surfactants. Although surfactants could substantially increase the amount of acropetal transport of radiolabel from both fungicides, none of those tested specifically promoted it; a constant proportion of the radioactive dose absorbed by a treated leaf was usually exported away from the site of application. The results are discussed in the light of current theories about the mode of action of surfactants as spray adjuvants.     

An Insecticidal and Acaricidal Polysulfide Metabolite from the Roots of Petiveria alliacea

The present study reports on the insecticidal and acaricidal potentials of dibenzyltrisulfide (DBTS) isolated from the roots of Petiveria alliacea L. using thin layer and high performance liquid chromatography. The 96-h LD₅₀ value (μg per tick) obtained for adult Boophilus microplus (Canestrini) topically treated with DBTS was 0·920. The LD₅₀ values obtained for three commercial acaricides dimethoate, lindane and carbaryl were 4·6, 9·3 and 6·9 μg per tick respectively. The IOD₅₀ and IHD₅₀ (concentrations inhibiting egg laying and hatching by 50% respectively) in μg per tick doses for DBTS were 0·22 and 0·24 respectively. The 24-h LD₅₀ dose (μg per insect) obtained for DBTS on adult Cylas formicarius elegantulus (Summer) was 0·193 μg per insect. The vapour from a stock solution of 5 g litre^-1 of DBTS was highly toxic to adult Hypothenemus hampei Ferr. inside coffee berries, inflicting 89% mortality within 24 h. © 1997 SCI.     

Leaching potential and decomposition of fluroxypyr in Swedish soils under field conditions

The mobility and decomposition of the herbicide fluroxypyr (4-amino-3,5-dichloro-6-fluoro-2-pyridyloxyacetic acid) was studied under field conditions in a sandy soil and a clay soil. Leachate was collected in lysimeters with undisturbed soil (sand) and in tile-drained plots (clay). Soil samples to a depth of one metre were also collected in both soils to characterize the temporal depth distribution of fluroxypyr in the profiles. The herbicide was applied as the I-methylheptyl ester of fluroxypyr at two rates, 187.5 and 375.0 g a.e. ha^?1, representing the normal and double the dose of the compound used for spring cereals. Some lysimeters received supplementary watering. Only two leachate samples (one from each soil) had concentrations of fluroxypyr above the detection limit (1 μg litre^?1), i.e. 2 and 5 μg litre^?1. Both samples were collected within two months after application, when less than 2 mm of drainage had been collected. The methylheptyl ester of fluroxypyr was not found in any of the samples. Fluroxypyr levels above the detection limit in soil (5 μg kg^?1 dry soil), were never found below the topsoil (0.2 m) in the clay profile, while, in the sandy profile, levels just above the detection limit were found occasionally in deeper soil layers. Concentrations were reduced to undetectable or very low levels within three months after spraying.     

乙草胺人工抗原的合成及其单克隆抗体的制备

通过乙草胺与3-巯基丙酸在碱性条件下反应合成了半抗原——乙草胺-巯基丙酸(AMPA)。采用活性酯法将半抗原AMPA分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联制备了人工抗原。通过紫外光谱测定了AMPA与BSA及OVA间的分子结合比分别为41和26。用AMPA-BSA免疫小鼠,制备得到的单克隆抗体的效价为1∶ 16×104。以AMPA-OVA作包被抗原,用乙草胺单克隆抗体建立了ELISA检测方法,IC50值为0.55 μg/L,方法的检测范围为0.04~5 μg/L,检测限为0.04 μg/L。     

Insecticidal activity of synthetic 2-carboxylbenzofurans and their coumarin precursors

Several 2-carboxylbenzofurans and their coumarin precursors, compounds 1 – 6 , and the novel compound 7 were synthesized and their insecticidal activity studied using adult sweet potato weevils (Cylas formicarius elegantulus (Summer)) at 24 μg, 48 μg and 72 μg per insect in 0.5 ml litre^?1 piperonyl butoxide (PPB) in acetone. The order of toxicity of the compounds as judged by the percentage mortality at 24 μg per insect at 48 h was 2 (100%) > 6 (95%) > 7 (85%) > 4 (75%)) > 3 (65%) > 1 (60%) > 5 (45%) > piperonyl butoxide (0.0%). Compound 2 was equitoxic to 24 μg of technical grade dimethoate. When administered in acetone alone, however, the compounds were only moderately active (i.e. > 120 μg per insect was required for 60% mortality at 48 h). The test compounds were more active at lower doses (48–72 μg per insect) when dissolved in dimethylsulfoxide or in an oil extract obtained from the fruit of Blighia sapida Koenig.     

Some physicochemical factors influencing foliar uptake enhancement of glyphosatemono(isopropylammonium) by polyoxyethylene surfactants

Structure-concentration–foliar uptake enhancement relationships between commercial polyoxyethylene primary aliphatic alcohol (A), nonylphenol (NP), primary aliphatic amine (AM) surfactants and the herbicide glyphosatemono(isopropylammonium) were studied in experiments with wheat (Triticum aestivum L.) and field bean (Vicia faba L.) plants growing under controlled-environment conditions. Candidate surfactants had mean molar ethylene oxide (EO) contents ranging from 5 to 20 and were added at concentrations varying from 0·2 to 10 g litre^?-1 to [¹⁴C]glyphosate formulations in acetone–water. Rates and total amounts of herbicide uptake from c. 0·2–μl droplet applications of formulations to leaves were influenced by surfactant EO content, surfactant hydrophobe composition, surfactant concentration, glyphosate concentration and plant species, in a complex manner. Surfactant effects were most pronounced at 0·5 g acid equivalent (a.e.) glyphosate litre^?-1 where, for both target species, surfactants of high EO content (15–20) were most effective at enhancing herbicide uptake: surfactants of lower EO content (5–10) frequently reduced, or failed to improve, glyphosate absorption. Whereas, at optimal EO content, AM surfactants caused greatest uptake enhancement on wheat, A surfactants gave the best overall performance on field bean; NP surfactants were generally the least efficient class of adjuvants on both species. Threshold concentrations of surfactants needed to increase glyphosate uptake were much higher in field bean than wheat (c. 2 g litre^?-1 and < 1 g litre^?-1, respectively); less herbicide was taken up by both species at high AM surfactant concentrations. At 5 and 10 g a.e. glyphosate litre^?-1, there were substantial increases in herbicide absorption and surfactant addition could cause effects on uptake that were different from those observed at lower herbicide doses. In particular, the influence of EO content on glyphosate uptake was now much less marked in both species, especially with AM surfactants. The fundamental importance of glyphosate concentration for its uptake was further emphasised by experiments using formulations with constant a.i./surfactant weight ratios. Any increased foliar penetration resulting from inclusion of surfactants in 0·5 g litre^?-1 [¹⁴C]glyphosate formulations gave concomitant increases in the amounts of radiolabel that were translocated away from the site of application. At these low herbicide doses, translocation of absorbed [¹⁴C]glyphosate in wheat was c. twice that in field bean; surfactant addition to the formulation did not increase the proportion transported in wheat but substantially enhanced it in field bean.     

Non-target effect of herbicides: I. effect of glyphosate and hexazinone on soil microbial activity. Microbial population,and in-vitro growth of ectomycorrhizal fungi

The effects of two herbicides, glyphosate (as a 359 g litre^?1 SL) and hexazinone (as a 50gkg^?1 granule) on soil microbial population, carbon dioxide evolution, and in-vitro growth of five species of ectomycorrhizal fungi were investigated. Glyphosate at 0–54 and 3.23 kg a.i. ha^?1 and hexazinone at 1. 2 and 8 kg a.i. ha^?1 did not reduce soil microbial population or carbon dioxide evolution in the long term (6 months). However, there was a significant short-term (2 months) effect of glyphosate on both fungal and bacterial counts at the 0.54 kg ha^?1 treatment. In in-vitro tests, Cenococcum graniforme. Hebeloma crustuliniforme and Laccaria laccata were more susceptible to both herbicides than was Suillus tomentosus. which was, in turn, more susceptible than Paxillus involutus. The growth of all five ectomycorrhizal fungi was significantly reduced when subjected to concentrations above 50 μl formulation litre^?1 (glyphosate) or 50 μg formulation litre^?1 (hexazinone).     

Development of Specific Recombinant Monoclonal Antibodies Against the Lipopolysaccharide of Ralstonia solanacearum Race 3

ABSTRACT Recombinant single-chain antibodies (scFvs) against the lipopolysaccharide of Ralstonia solanacearum (biovar 2, race 3) were successfully selected by phage display from a large combinatorial antibody library. Characterization with regard to cross-reaction and use in routine immunoassays showed that the selected antibodies had improved characteristics when compared with the polyclonal antiserum that is currently used for brown rot diagnosis of potato in the Netherlands. The isolated monoclonal scFvs reacted in both enzyme-linked immunosorbent assay (ELISA) and immunofluorescence cell staining with all race 3 strains tested, but with only some strains belonging to other races. Furthermore, only a few cross-reactions with saprophytic bacteria, which also cross-reacted with polyclonal antisera, were observed. Using ELISA, one of the recombinant antibodies detected as few as 5 x 10(3) bacteria in potato tuber extracts. Therefore, this antibody is potentially useful for detection of R. solanacearum race 3.