Inheritance of resistance to Bt canola in a field-derived population of Plutella xylostella

Crops expressing Bacillus thuringiensis (Bt) insecticidal Cry proteins are grown on millions of hectares. Recommendations to delay resistance are based on a high expression/refugia strategy that aims to kill resistant heterozygotes and enable some susceptible insects to survive. Leaf-dip bioassays on F1 crosses of Malaysian populations of diamondback moth (Plutella xylostella (L)) showed that CrylAc resistance was not fully recessive. The survival of ca 50% of heterozygotes on Bt canola (Brassica napus L) leaves expressing low concentrations of CrylAc agreed with a non-fully-recessive model for resistance. Extrapolations based on log dose-logit mortality regressions for heterozygotes using leaf-dip bioassays showed that a relatively high level of expression, of ca 2000 ng CrylAc mg(-1) total leaf protein, would be required to give 90% mortality to heterozygotes. If high enough levels of expression of Bt toxin to kill heterozygotes cannot be achieved and maintained under field conditions, the effectiveness of the high-dose/refugia strategy would be reduced.     

Sampling for the incidence of aphid-transmitted viruses in snap bean

ABSTRACT Data collected in 2002 and 2003 on Alfalfa mosaic virus and Cucumber mosaic virus incidences of infection in commercial snap bean fields in New York State were used to develop relationships between disease incidence (p(low)) and sample size while accounting for the inherent spatial aggregation of infected plants observed with these two viruses. For a plan consisting of 300 sampled plants (N = 60 quadrats, n = 5 plants per quadrat), estimating p(low) from the incidence of positive groups (p(high); testing of N = 60 grouped samples) provides the same precision in p(low) as testing 200 plants individually, up to about p(low) = 0.2. Above that, the confidence interval width for p(low) obtained via group testing becomes markedly larger than the width obtained by testing individual plants. Our results suggest using group testing until p(high) is in the range [0.35, 0.59], which corresponds to p(low) in [0.1, 0.2]. Results indicate that group testing can be more economical than the testing of individual plants without loss of precision, at lower incidences of infection. The approach presented provides a general framework for sampling and the estimation of incidence of other aphid-transmitted viruses in snap bean.     

Identification of immune response correlates for protection against bovine tuberculosis

Identification of an immune response correlate for protection against bovine tuberculosis would greatly facilitate the rational development of an effective vaccine. However, finding such a correlate has been a daunting task. Vaccination/challenge studies in cattle provide an ideal platform to compare induction of immune response parameters following vaccination and challenge, and assess the correlation of these parameters with protection. Protection against tuberculosis requires a Th 1-type cell-mediated immune response and induction of an antigen-specific interferon-gamma (IFN-gamma) response was the logical first choice in an investigation to identify an immune response correlate for protection. Calf vaccination studies showed that the subcutaneous injection of BCG vaccine induced significant protection against experimental challenge with Mycobacterium bovis. This protection was associated with strong whole blood IFN-gamma responses to bovine PPD 2-4 weeks after vaccination, but within the BCG-vaccinated groups, these responses were not correlated with protection. Use of a variety of vaccination strategies has shown that IFN-gamma responses in isolation were not necessarily associated with protection and concurrent IL-4 mRNA expression or antibody responses could also be induced. Collation of an immunological profile may be more informative than a study of individual cytokines. An indication of vaccine efficacy can be provided by the study of immune responses following challenge of the calves with M. bovis. IFN-gamma responses to ESAT-6, antibody responses following tuberculin skin testing and antigen-specific IL-4 mRNA expression all correlated with the severity of disease and indirectly provided an indication of protection. Future studies should be directed towards obtaining immunological profiles of calves following vaccination using techniques such as DNA microarray analysis, measurement of cytokine mRNA expression by real-time PCR, protein profiling by SELDI-TOF mass spectrometry as well as determining cytokine production by specific T cell sub-sets in individual protected animals.     

The bovine viral diarrhea virus (BVDV) NS3 protein, when expressed alone in mammalian cells, induces apoptosis which correlates with caspase-8 and caspase-9 activation

The bovine viral diarrhea virus (BVDV) strains exist as two biotypes, cytopathic (cp) and noncytopathic (ncp), according to their effects on tissue culture cells. It has been previously reported that cell death associated to cp BVDV in vitro is mediated by apoptosis. Here, experiments were conducted to determine the involvement of the NS3 protein in the induction of apoptosis. The NS3- and NS3Delta50 (deleted from the NH2-terminal 50 amino acids)-cDNA encoding sequences of BVDV NADL cp reference strain were cloned into adenoviral vectors (AdV) from which the BVDV gene of interest could be expressed from a tetracycline-responsive promoter. A549tTA cells infected in vitro with NS3 or NS3Delta50-expressing AdV showed cytopathic changes characterized by cell rounding and detachment, and nucleus chromatin condensation. DNA fragmentation assays, cytochrome c release, and activation of cellular caspases performed on these infected cells clearly correlated with the observed cytopathic changes with apoptosis. The BVDV NS3Delta50-induced apoptotic process was inhibited by caspase-8- and -9-specific peptide inhibitors (Z-IETD-FMK and Z-LEHD-FMK). Furthermore, apoptosis was inhibited in cells expressing the R1 subunit of herpes simplex virus type 2 ribonucleotide reductase (HSV2-R1) or hsp70, two proteins which are known to inhibit apoptosis associated with caspase-8 activation and cytochrome c release-dependent caspase-9 activation, respectively. Given that HSV2-R1, a specific inhibitor of the caspase-8 activation pathway, efficiently suppressed apoptosis and also prevented caspase-9 activation, the overall results indicate that the BVDV NS3/NS3Delta50 induces apoptosis initiated by caspase-8 activation and subsequent cytochrome c release-dependent caspase-9 activation.     

A New Natural Planthopper Vector of Stolbur Phytoplasma in the Genus Pentastiridius (Hemiptera: Cixiidae)

A new disease of sugar beet called Syndrome des Basses Richesses, which appeared in Burgundy and Franche-Comté, France, in 1991, is of uncertain aetiology. However, evidence for aerial transmission of the disease, symptom similarity with yellow wilt and preliminary results of phytoplasma detection, support the hypothesis of a phytoplasma being associated to the disease. A search for a natural phytoplasma vector, was conducted in Franche-Comté in 1997 and 1998, in an area where sugar beet crops had been affected since 1996. A cixiid, tentatively identified as Pentastiridius beieri, not described in the preceding years and not formerly reported as a phytoplasma vector, was present in sugar beet plots in high populations from June to August in 1997 and 1998. Individuals were captured and used for transmission experiments to periwinkle and sugar beet seedlings. They were further tested for the presence of a phytoplasma in their body, using PCR amplification of 16S rDNA of phytoplasmas. In 1997 and 1998, from 2% to 13.3% of the individuals carried a stolbur phytoplasma and insects which tested positive, appeared to have transmitted, through feeding, a stolbur phytoplasma to periwinkles and to sugar beets. This cixiid, whose vectoring capacity of stolbur phytoplasma to plants, is now clearly demonstrated, is available for experimental inoculations, in order to examine the role of phytoplasmas in the Syndrome des Basses Richesses, through the observation of symptom expression in phytoplasma-inoculated plants.     

Urea fertilization and foliar nutrient composition of western hemlock (Tsuga heterophylla (Raf.) Sarc.)

Foliar mineral contents of hemlock stands in coastal and Cascade Mountain zones of the Pacific Northwest were studied to determine the effect of urea fertilization on foliar nutrients. The results demonstrate that mineral concentrations vary among foliage age classes, stands, and zones. Concentrations of N, P, K, Ca, Mg, Al, Mn, Fe, Cu, Zn and B were significantly higher in the Cascade Mountain stands than in the coastal stands. Urea fertilization significantly increased the N content in all age classes of foliage but reduced the concentrations of P, Ca, Mg, Mn, Fe, Al and B, especially in coastal stands. The poor response of coastal stands to urea fertilization, therefore, is probably due to further reduction in already low foliar levels of essential nutrients other than N.     

A comparison of the efficiency of sprays of two drop-size ranges in an apple orchard

A two-season trial of a mist blower-sprayer with improved air-flow characteristics was made to assess the relative efficiency of 120 and 180 μm mass median diameter (MMD) drop-size sprays for the control of apple powdery mildew (Podosphaera leucotricha) and fruit tree red spider mite (Panonychus ulmi Koch) in a hedgerow-type apple orchard. The principal pesticide was binapacryl, applied at two rates for each drop-size. Under typical commercial conditions the smaller drop-size approximately halved the incidence of apple mildew found when the conventional 180 μm MMD drop-size spray was used; there was no conclusive advantage against red spider mite. Unexpectedly large variations occurred in the deposits from the weekly applications and were related to the humidity during spraying.     

A method for the estimation of 1,2-dibromoethane in vegetables

Residues of 1,2-dibromoethane (EDB) were readily estimated in treated vegetables down to levels of 0.2 mg/kg. Clean-up was effected by steam distillation into a toluene trap. This was followed by X-ray fluorescence determination of the collected EDB in the toluene. Good recoveries of added EDB were obtained at 0.5, 2, 5 and 20 mg/kg in cucumbers, pumpkins and zucchinis.     

Hemoperitoneum caused by the rupture of a granulosa cell tumor in a Holstein heifer

A case of a hemoperitoneum caused by the rupture of a granulosa cell tumor in a 9-month-old Holstein heifer is reported. Management of hemorrhagic shock in cattle is discussed.