Performance of soil-applied FeEDDHA isomers in delivering Fe to soybean plants in relation to the moment of application

FeEDDHA (iron(3+) ethylenediamine-N,N'-bis(hydroxyphenylacetic acid) products are commonly applied to mend and prevent Fe deficiency chlorosis in soil-grown crops. Plants mainly take up Fe in the progressed vegetative and in the reproductive stages. This study examined which of the principal constituents of FeEDDHA products (the isomers racemic o,o-FeEDDHA, meso o,o-FeEDDHA, and o,p-FeEDDHA), most effectively meets the Fe requirements of soybean plants (Glycine max (L.) Merr.) grown on calcareous soil in the aforementioned growth stages. FeEDDHA isomers were applied once, separately or in mixtures, at t = 0, in the progressed vegetative stage or in the reproductive stage. o,p-FeEDDHA did not significantly contribute to Fe uptake in either growth stage. Both racemic and meso o,o-FeEDDHA were effective in supplying plants with Fe, approximately to the same extent. The moment of application had a significant effect on yield and FeEDDHA pore water concentrations at harvest, but not on Fe uptake. To optimize yield while minimizing FeEDDHA dosage, FeEDDHA is best applied to soybean plants prior to the onset of chorosis.     

Monitoring Mycoplasma gallisepticum and Mycoplasma synoviae infection in breeder chickens after treatment with enrofloxacin.

Three experimental strains of breeder chickens were accidentally exposed to Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS), presumably from a newly introduced group of leghorn-type pullets. The experimental strains subsequently became infected and were diagnosed positive for MG and MS by the serum plate agglutination (SPA) test and confirmed by the hemagglutination inhibition (HI) test and the polymerase chain reaction (PCR) of tracheal swabs. Treatment with 10 mg/kg enrofloxacin via drinking water for 14 days was elected. Before and after initiation of treatment, MG and MS were monitored for changes by SPA, HI, PCR, and culture, with sampling intervals ranging from 1 wk to 7 wk. MG and MS SPA, HI, PCR, and culture were performed at each sampling period, with the exception of weeks 1.0 and 6.5. Week 1.0 included SPA and His for MG and MS. Week 6.5 included PCR and culture for MG and MS. The MG and MS SPA results were positive throughout the 29-wk trial period. MG HI titers declined until the last sampling, whereas the MS HI titers did not decline significantly. PCR for MG yielded only one positive result, which occurred before treatment. MS PCR remained positive throughout the trial period. MG was never isolated from any sample; however, one MS organism was isolated during treatment. The treatment regimen was effective for MG on the basis of PCR results. Treatment with enrofloxacin did not eliminate SPA reactions during the 29-wk trial period. MG HI titers remained in the suspicious range throughout the remainder of the trial period. Four weeks after the treatment ended, MG HIs were reduced by approximately 40%, with MS HIs remaining high throughout the 29-wk period. PCR appeared to be a sensitive and specific test on the basis of correlation with HIs. On the basis of the isolation of MS during treatment and continued subsequent PCR positive reactions, the treatment for MS with enrofloxacin was not as efficacious as for MG.     

Transmission of methicillin resistant Staphylococcus aureus among pigs during transportation from farm to abattoir

The prevalence of methicillin resistant Staphylococcus aureus (MRSA) in pigs at abattoirs is higher than in pigs sampled on farms. This study investigated whether MRSA negative pigs can become MRSA positive during transportation from the farm to the abattoir after exposure to other pigs and environmental sources of MRSA. Nasal swabs were collected from four batches of pigs during loading at the farm, on arrival at the abattoir and after stunning. Environmental wipes were taken from lorries after transporting pigs and from lairages after holding pigs. All pigs (n = 117) tested MRSA negative before transportation. On arrival at the abattoir, 12/117 (10.3%) pigs in two batches tested MRSA positive. In lorries that tested positive after transportation, the prevalence of MRSA positive pigs was 21.1%, whereas no MRSA was detected in pigs that had been transported in lorries that tested negative after transportation. At stunning, all batches and 70/117 (59.8%) pigs tested MRSA positive. Pigs can become MRSA positive in the short period of time during transportation from the farm to stunning at the abattoir.     

Epitope identification and <Emphasis Type="Italic">in silico</Emphasis> prediction of the specificity of antibodies binding to the coat proteins of <Emphasis Type="Italic">Potato Virus Y</Emphasis> strains

A phage library containing 2.7 × 10⁹ randomly expressed peptides was used to determine the epitopes of three monoclonal antibodies that bind to the coat protein of potato virus Y. Construction of the consensus sequences for the peptides obtained after three selection rounds indicated that each antibody recognized a different epitope located within the first 50 N-terminal amino acids of the coat protein. The location of the epitopes was confirmed by heterologous expression of the N-terminal part of the coat protein in Escherichia coli, and, subsequently, by performing an immunological test with the three antibodies. The accuracy of the phage library was demonstrated by predicting in silico the cross-reactivity of the three antibodies with other potyvirus family members. ELISA and in silico predictions revealed the same results in almost every case. The potential of peptide phage libraries to optimize the use of antibodies in plant virology is discussed.     

Display and selection of chicken IgA Fab fragments

Passive immune therapy is regaining interest to prevent and cure infectious diseases both in human and veterinary medicine. Therefore, systems are required that enable efficient targeted selection of antibodies originating from virtually any animal species. Here, a system for the selection of chicken IgA, using phage display, is described. A novel phagemid vector (pChick3) for the display and selection of chicken IgA antibodies in Fab format was developed. The functionality of pChick3 was demonstrated by construction of an immune antibody library using B cells from chickens infected with Eimeria acervulina. From this library, 10 different IgA fragments with specific binding to the E. acervulina antigen mix, the sporozoite or oocyst fractions were selected. These results demonstrate the efficiency and versatility of the pChick3 vector system that can readily be applied to construct libraries and subsequently select antibodies of the alpha isotype against a wide variety of pathogens and parasites.     

Quantification of transmission of livestock-associated methicillin resistant Staphylococcus aureus in pigs

Antimicrobial resistance in pigs becomes a public health issue when resistant organisms transfer from pigs to humans. Pigs are a large reservoir for livestock-associated (LA-)MRSA and people in contact with pigs are at risk for infection with LA-MRSA. Transmission and persistence of LA-MRSA within a pig population contributes to the maintenance of this zoonotic reservoir. Current knowledge on colonization and transmission of LA-MRSA in pigs is limited and mainly based on observational field surveys. Two experiments were performed to colonize pigs and quantify transmission of LA-MRSA between pigs. In the first experiment, colonization of six-week old piglets failed after intranasal inoculation, confirming the complexity of MRSA-colonization. In the second experiment, naive pigs got colonized after exposure to orally inoculated pigs. Subsequently, these contact-infected pigs transmitted MRSA to a new group of naive pigs. The reproduction ratio, R(0), was estimated with a SIS-model to quantify transmission between the first and second contact pigs as this resembles more the natural transmission. Two scenarios were evaluated, with different assumptions regarding infection status of individual pigs. R(0) varied between 3.7 and 4.3 and was significantly above 1, indicating a high probability of persistence of LA-MRSA, even without antimicrobial use.     

Stable recombinant alpaca antibodies for detection of Tulip virus X

For detection of the plant pathogenic Tulip virus X (TuVX), a panel of six recombinant antibodies was identified. To this end, a repertoire of variable domains from heavy-chain immunoglobulins (VHH) was cloned from an alpaca, which had been immunized with TuVX. Binding domains were selected by phage display and panning on immobilized TuVX particles. Recombinant VHH antibodies were tested for sensitivity in a sandwich ELISA, and were demonstrated to be readily able to distinguish TuVX-infected tulip leaf material from uninfected leafs. No cross-reactivity of the VHH antibodies to related flexiviridae was observed. Recombinant VHHs maintained their reactivity upon storage at −20°C for over a year. The effect of incubation at higher temperatures for prolonged time was studied. Two out of three VHH proteins retained activity after several weeks of storage at 37°C.     

Quantitative resistance to Botrytis cinerea from Solanum neorickii

Tomato (Solanum lycopersicum) is susceptible to gray mold (Botrytis cinerea). Quantitative resistance to B. cinerea was previously identified in a wild relative, S. neorickii G1.1601. The 122 F₃ families derived from a cross between the susceptible S. lycopersicum cv. Moneymaker and the partially resistant S. neorickii G1.1601 were tested for susceptibility to B. cinerea using a stem bioassay. Three putative quantitative trait loci (pQTL) were detected: pQTL3 and pQTL9 reducing lesion growth (LG) and pQTL4 reducing disease incidence (DI). For each pQTL, a putative homologous locus was identified recently in another wild tomato relative, S. habrochaites LYC4. pQTL3 was confirmed by assessing disease resistance in BC₃S₁ and BC₃S₂ progenies of S. neorickii G1.1601. pQTL4 was not statistically confirmed but the presence of the S. neorickii resistance allele reduced DI in all three tested populations. The reduction in LG of pQTL9 was not confirmed but rather, this locus conferred a reduced DI, similar to observations in the QTL study using S. habrochaites. The results are discussed in relation to other disease resistance loci identified in studies with other wild tomato relatives.     

A major genome region underlying artemisinin resistance in malaria

Evolving resistance to artemisinin-based compounds threatens to derail attempts to control malaria. Resistance has been confirmed in western Cambodia and has recently emerged in western Thailand, but is absent from neighboring Laos. Artemisinin resistance results in reduced parasite clearance rates (CRs) after treatment. We used a two-phase strategy to identify genome region(s) underlying this ongoing selective event. Geographical differentiation and haplotype structure at 6969 polymorphic single-nucleotide polymorphisms (SNPs) in 91 parasites from Cambodia, Thailand, and Laos identified 33 genome regions under strong selection. We screened SNPs and microsatellites within these regions in 715 parasites from Thailand, identifying a selective sweep on chromosome 13 that shows strong association (P = 10(-6) to 10(-12)) with slow CRs, illustrating the efficacy of targeted association for identifying the genetic basis of adaptive traits.