Enzyme-linked Immunosorbent Assay Absorbance Research Articles

The susceptibility and response of inbred and outbred Mimulus guttatus to infection by Cucumber mosaic virus

The deleterious effects of inbreeding have been well documented, but only recently have studies begun to explore the consequences of inbreeding for important ecological interactions. We examined the effects of inbreeding on the interaction between host and pathogen using the mixed-mating Mimulus guttatus (Scrophulariaceae) and Cucumber mosaic virus (CMV). Inbred (self) and outbred M. guttatus from two California populations (M5 and M13) were rub-inoculated with CMV and compared to sham-inoculated controls. Flower production by outbred plants in host population M5 showed little effect of the inoculation treatment, but inoculation reduced flower production of inbred plants by 12%, indicating that inbreeding reduces tolerance to CMV infection. This interaction fell short of significance, however. The effects of inbreeding and CMV inoculation on biomass in M5 varied significantly across the 15 families used in this experiment, indicating genetic variation in the effect of inbreeding on resistance or tolerance to CMV. CMV infection reduced biomass in host population M13, but there were no significant interactions between virus treatment and level of inbreeding for either flower production or biomass. Enzyme linked immunosorbent assay (ELISA) was used to detect CMV in host tissues. In both populations, mean ELISA absorbance values of inoculated plants were nearly identical for self and outcross hosts, indicating equal susceptibility to CMV. In outbred plants of population M5, flower production did not change with increasing ELISA absorbance, but in inbred plants it declined, indicating reduced tolerance to CMV infection. The results from this study suggest that pathogens may become increasingly detrimental as host populations become more inbred.

Apoptosis enzyme-linked immunosorbent assay distinguishes anticancer drugs from toxic chemicals and predicts drug synergism

The effects of anticancer drugs and toxic compounds on leukemic cells in culture were evaluated by enzyme-linked-immunosorbent assay (ELISA) based on the detection of apoptotic cells by a monoclonal antibody against single-stranded DNA. The concentrations of 13 anticancer drugs, which increased apoptosis ELISA absorbance, were similar to the concentrations decreasing long-term cell survival. Short-term metabolic tetrazolium-based 3-(4,5-dimethylthiazol-yl)-2,5-diphenyformazan bromide (MTT) assay was significantly less sensitive than apoptosis ELISA and the cell survival assay. In contrast to anticancer drugs, 12 toxic chemicals did not increase apoptosis ELISA absorbance at cytotoxic concentrations. The difference between two groups of compounds by apoptosis ELISA was especially large in cultures treated with twofold of concentrations producing 50% inhibition of cell growth: all anticancer drugs induced intense reaction (mean absorbance 2.0), while none of the toxic chemicals induced apoptosis. The application of apoptosis ELISA to chemosensitivity testing was evaluated by its ability to detect synergism of anticancer drug combinations. Among 66 drug combinations tested, only combination of nitrogen mustard with mithramycin was highly synergistic by the apoptosis ELISA, as defined by apoptosis induction with the combination containing each drug at 50% of effective concentration. This combination was also synergistic in the cell survival assay, producing significant cell kill while each drug alone had no effect on cell survival. This synergism was not detected by MTT assay. We conclude that apoptosis ELISA could be useful for drug development and chemosensitivity assessment as it can distinguish clinically useful anticancer drugs from toxic compounds, is as sensitive as the long-term cell survival assay and can detect anticancer drug synergism by rapid evaluation of apoptosis induction.

Detection and estimation of avian infectious bronchitis virus antigen by a novel indirect liquid-phase blocking enzyme-linked immunosorbent assay using chicken and rabbit affinity purified immunoglobulins

An indirect liquid-phase blocking (LPB) enzyme-linked immunosorbent assay (ELISA) using chicken and rabbit affinity purified immunoglobulin G (IgG) has been developed to detect and estimate avian infectious bronchitis virus (IBV) antigen concentration directly in infected allantoic fluid. The method is based on the principle of binding of specific IgG to the test IBV antigen and the assay of unbound IgG on an antigen-coated ELISA plate. The immunoglobulins are chicken N-terminal S2 peplomeric protein-specific IgG isolated by immunoaffinity chromatography on synthetic peptide coupled to CNBr-activated Sepharose 4B or rabbit polyclonal IgG purified from the serum using Protein A Sepharose 4B. The assay detected all tested IBV strains and field isolates propagated in chicken embryos. Signal to noise ratios were calculated from LPB ELISA absorbance units and a diagnostic threshold was established from the signal to noise ratio frequency distribution of samples positive or negative for IBV by virus titration or reverse transcription polymerase chain reaction. The relative sensitivity of the test ranged between 10 5 and 10 6 median egg infectious doses (EID 50 ) for chicken IgG and between 10 3 and 10 4 EID 50 for rabbit IgG, depending on the test strain. The assay is simple and takes less than 3 h to perform. It does not require expensive reagents and can be readily adapted to monitor the IBV antigen concentration in allantoic fluids during propagation of vaccine strains or in samples of freeze-dried, live-attenuated IBV vaccines.

Enzyme-linked immunosorbent assay (ELISA) for the specific detection of apoptotic cells and its application to rapid drug screening.

We have developed a solid-phase ELISA for the specific and sensitive detection of apoptotic cells. This method is based on the ability of a monoclonal antibody (MAb) against single-stranded DNA (ssDNA) to specifically identify apoptotic cells. The assay involves binding of cells to 96-well microtiter plates, treatment of the attached cells with formamide to denature DNA in apoptotic cells and one-step staining of the denatured DNA with a mixture of anti-ssDNA MAb and peroxidase-conjugated anti-mouse IgM. A near linear increase in signal was seen as the number of apoptotic cells increased from 500 to 5000. Untreated and necrotic cells or cells with single-stranded DNA breaks induced by H 2O 2 did not produce signal above the background level. In leukemic cell cultures grown, treated with ID 50 concentration of etoposide, stained and analyzed in the same 96-well assay plate, intense ELISA signal was detected. The ratio of absorbance values from drug resistant and drug-sensitive cell lines treated with etoposide was in agreement with the degree of resistance determined by growth inhibition assays. These data show that this ELISA has sufficient sensitivity for use in drug screening protocols. In breast cancer cell cultures treated with cisplatin, ELISA absorbance increased only after treatment with drug concentrations 10-fold higher than concentrations inducing 95% growth inhibition. In cultures treated with staurosporine, there was a near linear relation between the ELISA absorbance values and cytotoxicity in the range of 15–92% growth inhibition. The absence of apoptotic signal in breast cancer cells treated with cytotoxic concentrations of cisplatin indicated that this drug kills cells by non-apoptotic mechanisms, whereas apoptosis was the dominant mechanism of cell death caused by staurosporine. The formamide-MAb apoptosis ELISA described here may provide a basis for high-throughput screening of drugs based on their ability to induce or suppress apoptosis.

Immunoprophylaxis AgainstFasciola hepaticain Rabbits Using a Recombinant Fh15 Fatty Acid-Binding Protein

Previous studies of ours have demonstrated that a recombinant protein (Fh15) related to fatty acid-binding proteins did not induce significant protection in rabbits challenged 2 or 4 wk postimmunization over nonimmunized controls. In the current study, rabbits were immunized with Fh15 and challenged with Fasciola hepatica metacercariae 12 and 20 wk later. In the current study in which longer lag periods for challenge infection after the second immunization were used, worm burden reductions compared to adjuvant controls were a significant 43% and 76%, respectively. Importantly, rabbits immunized with Fh15 had significant numbers of immature flukes, 66% in the 12-wk period and 84% in the 20-wk lag period as compared to controls. In addition, liver lesions were clearly diminished in the vaccinated rabbits. Enzyme-linked immunosorbent assay absorbance values showed that immunized rabbits developed high antibody levels to Fh15 from 8 wk after the first immunization and did not increase after challenge. These results suggest that a recombinant F. hepatica molecule related to fatty acid-binding proteins induces protective (worm burden reductions), anti-fecundity (immature flukes), and anti-pathology (less liver lesions) effects in rabbits and may serve as a model for the immunoprophylaxis of fascioliasis.

Haemophilus somnus-induced IgE in calves vaccinated with commercial monovalent H. somnus bacterins.

The ability of commercially available Haemophilus somnus bacterins to elicit an immunoglobulin E (IgE) response was examined in healthy calves using enzyme-linked immunosorbent assay (ELISA) and western blotting techniques. Thirty five calves were utilized in this study. Calves in Group 1 (n=7) did not receive any H. somnus vaccination and served as negative controls. Calves in each of Groups 2-5 (n=7 each) were vaccinated on days 0 (primary) and 14 (booster) with one of four commercially available H. somnus bacterins. Sera were harvested on days 0 and 14 and at weekly intervals for a total of 45 days. Sera were tested for the presence of IgE antibodies using a bovine IgE-specific ELISA. Low levels of H. somnus-specific IgE were detected by ELISA in all animals prior to the initiation of the study. All bacterins induced IgE levels that were significantly higher than control levels. Two bacterins elicited higher IgE levels at all time points. Sera were adsorbed against washed whole cells of either Salmonella typhimurium, P. multocida, or H. somnus or extracts of H. somnus. ELISA absorbance values were significantly decreased by adsorption with washed whole cells or extracts of H. somnus, whereas adsorption with other gram-negative bacteria only minimally decreased ELISA absorbance values. These results indicate that commercially available H. somnus bacterins can induce IgE antibody as early as 14 days post-vaccination. This IgE can be detected 45 days after the primary vaccination. Results also indicate that H. somnus-specific IgE antibodies can be found in serum of some cattle, possibly induced by existing or previous sensitization.

New Source of Resistance to Cucumber mosaic virus in Capsicum frutescens.

A small-fruited pungent pepper accession, Capsicum frutescens 'BG2814-6', is resistant to several isolates of Cucumber mosaic virus (CMV). Resistance in BG2814-6 is incompletely penetrant and is controlled by at least two major recessive genes. The accession BG2814-6 and C. annuum 'Perennial', the leading source of CMV tolerance, appear to share one or more CMV resistance genes. CMV was detected in uninoculated leaves in a small percentage of both BG2814-6 and Perennial plants, indicating that resistance is not absolute in either genotype. Enzyme-linked immunosorbent assay absorbance values of samples taken from inoculated leaves corresponded well with visible viral symptoms for parental genotypes. While Perennial plants accumulated CMV antigen in inoculated leaves, CMV antigen was not detected in inoculated leaves of 73% of BG2814-6 plants, suggesting that there may be a mechanistic difference in resistance between the two genotypes.

Evaluation of serodiagnosis of toxoplasmosis by using the recombinant nucleoside triphosphate hydrolase isoforms expressed in Escherichia coli

The nucleoside triphosphate hydrolase (NTPase) isoforms termed, NTPase-I and NTPase-II of Toxoplasma gondii, were expressed in Escherichia coli as inclusion bodies and purified under denaturing condition. Furthermore, NTPase-I was refolded as an active form and purified under non-denaturing condition. The purified NTPase isoforms, both denatured and refolded, were tested for their usefulness as antigens for the serodiagnosis of acute toxoplasmosis in immunocompetent humans. The test was conducted by using the recombinant NTPase isoforms and comparing the enzyme linked immunosorbent assay (ELISA) absorbances with the Sabin–Feldman dye test titer. Seventy-three sera from dye test-positive patients, and 30 sera from subjects with no T. gondii infection were examined. The total positive rates in dye test positive sera were: 82% (60/73) for denatured NTPase-I; 78% (57/73) for denatured NTPase-II; and 63% (46/73) for refolded NTPase-I. For all three antigen types of recombinant NTPase, the positive rates of sera of acute toxoplasmosis suspected patients were 93% (13/14). A moderate correlation between the ELISA absorbance using these antigens and the dye test titer was observed with the correlation coefficients, 0.583 ( r 2) for denatured NTPase-I, 0.472 ( r 2) for denatured NTPase-II, and 0.604 ( r 2) for refolded NTPase-I in the linear regression analysis. There was no significant difference observed in the antigenicity between refolded and denatured NTPase-I, nor between the isoforms.

Detection of cucumber mosaic cucumovirus in weed species: a cautionary report on nonspecific reactions in ELISA

The reliability of enzyme-linked immunosorbent assay (ELISA) to detect cucumber mosaic cucumovirus (CMV) coat protein (CP) in three weed species (Taraxacum officinale, Gnaphalium obtusifolium, and Conyza canadensis) was compared with other detection methods, including Western blot analysis and a bioassay. Analysis of extracts prepared from these species by double antibody sandwich (DAS) ELISA yielded high absorbance values (A4O5nm) that were indicative of CMV concentrations of purified CMV standards of greater than 50 mg/mL. However, results from an ELISA procedure designed such that no viral antigen should be detected suggested that the strong ELISA absorbance values were nonspecific reactions with material(s) in the weed extracts. These findings were substantiated when no CMV CP was detected by Western blot analysis or infectious CMV was detected in bioassays involving inoculation of extracts onto an indicator host (Cucurbita pepo ‘Crookneck’ (squash)). In both DAS ELISA and indirect ELISA procedures, the nonspecific reaction was shown to result from the interaction of weed extract and alkaline phosphatase-conjugated immunoglobulin (including anti-CMV and goat anti-rabbit). Addition of bovine serum albumin to the extraction buffer for DAS ELISA or to the extraction buffer and at each antibody step for indirect ELISA dramatically reduced the nonspecific reactions.

The Genetic Basis of High Resistance to Rice Yellow Mottle Virus (RYMV) in Cultivars of Two Cultivated Rice Species.

Three cultivars of Oryza sativa (IR64, Azucena, and Gigante) and four cultivars of O. glaberrima (Tog5681, Tog5673, CG14, and SG329) were evaluated for their resistance to two isolates of rice yellow mottle virus (RYMV) by enzyme-linked immunosorbent assay (ELISA) and symptomatology. Cultivars Tog5681 and Gigante were highly resistant, and no symptoms were observed when either virus isolate was inoculated at 10 or 20 days postgermination and assayed by ELISA at 7, 14, 22, 35, 50, or 64 days postinoculation. Azucena showed a partial resistance, whereas the other cultivars were susceptible. Symptom appearance was associated with increase in ELISA absorbance in the systemically infected leaves. The best discrimination among the cultivars occurred when the plants were inoculated at 10 days postgermination. Crosses were made between the highly resistant (Gigante and Tog5681) and the susceptible (IR64) cultivars to determine the genetic basis of resistance to RYMV. Evaluation of F1 hybrids and interspecific progenies, as well as the segregation of resistance in F2 and F3 lines of the IR64 × Gigante cross, provided results consistent with the presence of a single recessive resistance gene common to Tog5681 and Gigante.

Development of a Field Enzyme‐Linked Immunosorbent Assay (ELISA) for Detection of α‐Amylase in Preharvest‐Sprouted Wheat

ABSTRACTA sandwich enzyme‐linked immunosorbent assay (ELISA) has been developed for detection of α‐amylase in preharvest sprouted wheat and adapted to rapid field‐use formats requiring 15–20 min to perform. Polyclonal and monoclonal antibodies were prepared to detect a mixture of high and low pI isozymes of α‐amylase and high pI isozymes only. All antibodies detected α‐amylase on immunoblots of either a crude wheat extract or of purified enzyme, but only the polyclonal antibodies functioned in a sandwich ELISA. Depending on the antibody combination, the tube ELISA detected either the high and low pI isozymes of α‐amylase or the high pI isozymes only with a detection limit of ≈0.5–1.0 ng/mL of amylase. Wheats with falling numbers (FN) of <350 sec could be discriminated from sound wheats, with decreasing FN producing increasing assay color. Using 130 wheat grain samples, ELISA absorbances for detection of both high and low pI isozymes and of high pI isozymes only were highly positively correlated with amylase enzyme activity and negatively correlated with FN. The correlations were similar for detection of both isozyme families and for detection of high pI isozymes only. Analyses of three sets of wheat samples from different environments demonstrated that the relationship between ELISA absorbance and FN had little dependence on wheat cultivar. The precision of sample analysis using the field ELISA was similar to the precision of FN test apparatus.

Chlamydia pneumoniae infection of the central nervous system in multiple sclerosis

Our identification of Chlamydia pneumoniae in the cerebrospinal fluid (CSF) of a patient with multiple sclerosis (MS) led us to examine the incidence of this organism in the CSF from 17 patients with relapsing-remitting MS, 20 patients with progressive MS, and 27 patients with other neurological diseases (OND). CSF samples were examined for C pneumoniae by culture, polymerase chain reaction assays, and CSF immunoglobulin (Ig) reactivity with C pneumoniae elementary body antigens. C pneumoniae was isolated from CSF in 64% of MS patients versus 11% of OND controls. Polymerase chain reaction assays demonstrated the presence of C pneumoniae MOMP gene in the CSF of 97% of MS patients versus 18% of OND controls. Finally, 86% of MS patients had increased CSF antibodies to C pneumoniae elementary body antigens as shown by enzyme-linked immunosorbent assay absorbance values that were 3 SD greater than those seen in OND controls. The specificity of this antibody response was confirmed by western blot assays of the CSF, using elementary body antigens. Moreover, CSF isoelectric focusing followed by western blot assays revealed cationic antibodies against C pneumoniae. Infection of the central nervous system with C pneumoniae is a frequent occurrence in MS patients. Although the organism could represent the pathogenetic agent of MS, it may simply represent a secondary infection of damaged central nervous system tissue. A therapeutic trial directed at eliminating C pneumoniae from the central nervous system may provide additional information on its role in MS.

Chlamydia pneumoniae infection of the central nervous system in multiple sclerosis

Our identification of Chlamydia pneumoniae in the cerebrospinal fluid (CSF) of a patient with multiple sclerosis (MS) led us to examine the incidence of this organism in the CSF from 17 patients with relapsing–remitting MS, 20 patients with progressive MS, and 27 patients with other neurological diseases (OND). CSF samples were examined for C pneumoniae by culture, polymerase chain reaction assays, and CSF immunoglobulin (Ig) reactivity with C pneumoniae elementary body antigens. C pneumoniae was isolated from CSF in 64% of MS patients versus 11% of OND controls. Polymerase chain reaction assays demonstrated the presence of C pneumoniae MOMP gene in the CSF of 97% of MS patients versus 18% of OND controls. Finally, 86% of MS patients had increased CSF antibodies to C pneumoniae elementary body antigens as shown by enzyme-linked immunosorbent assay absorbance values that were 3 SD greater than those seen in OND controls. The specificity of this antibody response was confirmed by western blot assays of the CSF, using elementary body antigens. Moreover, CSF isoelectric focusing followed by western blot assays revealed cationic antibodies against C pneumoniae. Infection of the central nervous system with C pneumoniae is a frequent occurrence in MS patients. Although the organism could represent the pathogenetic agent of MS, it may simply represent a secondary infection of damaged central nervous system tissue. A therapeutic trial directed at eliminating C pneumoniae from the central nervous system may provide additional information on its role in MS. Ann Neurol 1999;46:6–14

Influence of Infection withRenibacterium salmoninarumon Susceptibility of Juvenile Spring Chinook Salmon to Gas Bubble Trauma

Abstract During experiments in our laboratory to assess the progression and severity of gas bubble trauma (GBT) in juvenile spring chinook salmon Oncorhynchus tshawytscha, we had the opportunity to assess the influence of Renibacterium salmoninarum (Rs), the causative agent of bacterial kidney disease, on the susceptibility of salmon to GBT. We exposed fish with an established infection of Rs to 120% total dissolved gas (TDG) for 96 h and monitored severity of GBT signs in the fins and gills, Rs infection level in kidneys by using an enzyme-linked immunosorbent assay (ELISA), and mortality. Mortality occurred rapidly after exposure to 120% TDG, with a LT20 (time necessary to kill 20% of the population) of about 37 h, which is at a minimum about 16% earlier than other bioassays we have conducted using fish that had no apparent signs of disease. Fish that died early (from 31 to 36 h and from 49 to 52 h) had significantly higher infection levels (mean ± SE ELISA absorbance = 1.532 ± 0.108) than fish that sur...

Disparity between serological reactivity to Borrelia burgdorferi and evidence of past disease in a high-risk group.

A prevalence study of past Lyme borreliosis in persons with outdoor occupations was done. Consenting individuals (n = 302) were administered a questionnaire eliciting demographic and occupational data and a clinical history, and were asked to donate a serum specimen for detection of antibodies to Borrelia burgdorferi by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and borrelia inhibition assays, and for detection of potentially cross-reactive antibodies. Of 302 individuals, 77 (25%) had reactive antibodies detected by ELISA. Of these 302 individuals, 44 (15%) met the criteria of the Centers for Disease Control and Prevention for serological reactivity as evidenced by immunoblotting, and 70 (23%) had inhibitory activity. Through the clinical criteria employed, only 11 individuals with serological reactivity had prior illness compatible with Lyme borreliosis. Higher ELISA absorbances were positively correlated with age and duration of outdoor occupation. The results from three serological assays and the lack of reactivity to potentially cross-reactive infectious agents indicate that serological reactivity was due to exposure to B. burgdorferi. The disparity between serological reactivity and the clinical evidence of Lyme borreliosis suggests cumulative exposure to a nonpathogenic form of B. burgdorferi.

Antigenic variation in Bacteroides forsythus detected by a checkerboard enzyme-linked immunosorbent assay.

Evidence indicating that multiple serotypes of Bacteroides forsythus participate in rapidly progressing periodontal infections has not been reported previously. Our aim was to develop an assay for detecting subsets of B. forsythus clinical isolates which differ in serogroup membership and subsets of patients with immunoglobulin G (IgG) responses which differ in serogroup recognition. A checkerboard enzyme-linked immunosorbent assay (ELISA) was used to assess variation in the IgG binding profiles of 22 clinical isolates in sera from 28 patients with early-onset rapidly progressive periodontitis. To accommodate the maximum number of isolates and sera in a given assay run, a multiplate assay grid with standard 96-well microtest plates was established. Single dilutions of individual sera were placed in rows crossing columns of isolate-coated wells, and antigen-specific IgG immobilized in the wells was measured as ELISA absorbance. Pooled sera and isolates were assayed in parallel to serve as negative controls for variation in IgG binding profiles. Correlation and hierarchical cluster analysis of the absorbance data matrix showed that the isolates could be sorted into at least four clusters based on variations in their IgG binding profiles across different sera. Furthermore, at least two patient clusters were defined by variations in their serum IgG antigen recognition profiles across different isolates. We conclude that multiple serogroups of B. forsythus exist and that different serogroups are dominant in the antibody response of different patients. The method applied here could be used to serologically classify clinical isolates of other species which evoke a serum antibody response in patients.

Detection of Antibody against Lymphocystis Disease Virus in Japanese Flounder by Enzyme Linked Immunosorbent Assay.

Enzyme linked immunosorbent assay (ELISA) was applied to detect the antibody of Japanese flounder (Paralichthys olivaceus) against lymphocystis disease virus (LCDV). Purified Japanese flounder LCDV was used as a capture antigen of ELISA. Monitoring the immune response of Japanese flounder to LCDV was done using the sera obtained from apparently healthy, LCD-diseased and LCD-recovered fish. ELISA absorbances of these three groups were clearly different. Using the serum with high ELISA absorbance, the optimum ELISA condition was set up. Apparently healthy fish were injected with inactivated LCDV. One to three months after the injection, the antibody against LCDV was detected by ELISA established for LCDV and their ELISA antibody titer increased during that period.

Identification of the Major Epitope in the GL Protein of Equine Arteritis Virus (EAV) Recognized by Antibody in EAV-infected Horses Using Synthetic Peptides.

A panel of overlapping peptides covering the ectodomain of the GL protein of equine arteritis virus (EAV) was synthesized to identify the major epitope recognized by the EAV-seropositive horse sera by using enzyme-linked immunosorbent assay (ELISA). Three consecutive peptides, G15 (amino acid residues 75-90), G16 (79-94) and G17 (83-98), strongly reacted with 3 experimentally EAV-infected horse sera. G16 peptide was highly antigenic with all 8 EAV-infected horse sera tested. The ELISA absorbance values and the virus neutralization titers of antibodies in sera periodically collected from EAV-infected horses showed similar patterns. These results indicated that the major linear epitope of the EAV GL protein which was recognized with EAV-infected horse sera was mapped to residues from 83 to 90.

Enzyme-linked immunosorbent assay for serological survey of equine arteritis virus in racehorses.

To examine antibodies against equine arteritis virus (EAV), an enzyme-linked immunosorbent assay (ELISA) using purified virus antigen was developed. The results of ELISA were compared with those of serum neutralization (SN) tests. The ELISA absorbance values and the SN titers in sera collected weekly from EAV-infected horses showed a similar pattern. The ELISA could detect antibody to EAV in horses experimentally infected with not only a homologous virus strain, which was used as the ELISA antigen, but also a heterologous strain. Using the ELISA, serum samples collected in 1996 from racehorses in three prefectures (Hokkaido, Ibaraki, and Shiga) were examined and there was no evidence of recent EAV infection among these racehorse populations in Japan. The ELISA should be a simple and highly specific method for rapid screening of EAV infection in racehorses.

Lack of Association between Fusarium Foot Rot and Head Blight Resistance in Winter Rye

Foot rot and head blight caused by Fusarium culmorum (W.G. Sm.) Sacc. may lead to considerable loss of yield and grain quality in winter rye (Secale cereale L.). Breeding for resistance to the two diseases would be facilitated if the same resistance mechanisms were involved. The purpose of this study was to estimate the genetic correlation between the resistance to the two diseases in advanced winter rye breeding materials. Twenty self‐fertile inbred lines were grown at two locations in South Germany in 1992, 1993, and 1994 (six environments). Lines were arranged in three adjacent treatment blocks: no inoculation, inoculation for foot rot, and inoculation for head blight. Disease severity was rated on a 1‐to‐9 scale for foot rot and head blight. Besides the foot rot and head blight rating scale, ELISA (enzyme‐linked immunosorbent assay) was used to quantify the amount of Fusarium protein within stem issue from the foot rot treatment and relative grain weight was measured from the head blight treatment. In all but one environment, F. culmorum was re‐isolated from diseased stems at high percentages (≥90%). Disease severity was moderate for foot rot and head blight rating at all environments. Mean ELISA absorbance was 0.55 and mean relative grain weight 77.4%. Significant genotypic variation existed for all resistance traits. Broad‐sense heritabilities, calculated on an entry‐mean basis, were high for foot rot and head blight rating (h2 ≈ 0.8) and somewhat lower for ELISA absorbance and relative grain weight (h2 ≈ 0.5–0.6). No significant correlation was found between the resistances to foot rot and head blight. This was confirmed by a highly significant (P = 0.01) genotype‐plant organ interaction. Thus, resistance mechanisms most likely differ for the two diseases. Resistance selection requires separate multi‐environmental screening tests for F. culmorum foot rot and head blight.