An assay for quantitative virulence in Rhynchosporium commune reveals an association between effector genotype and virulence

Detailed knowledge of the evolutionary genetics of virulence is needed to understand and predict host–pathogen dynamics. This study used a virulence assay based on digital image analysis and treated virulence as a quantitative rather than a binary trait. Such quantitative data may better reflect the genetic underpinning of virulence in many pathogen systems and provide better resolution in statistical investigations. A greenhouse experiment based on a common garden design was conducted to measure virulence (% of leaf area covered by lesions) of 126 genetically distinct isolates of the barley scald pathogen, Rhynchosporium commune, originating from nine field populations around the world. Virulence in this pathosystem was found to be a quantitative trait with a continuous distribution in all populations. By comparing population genetic differentiation for virulence and neutral microsatellite markers (i.e. a Q_ST/G_ST comparison), evidence that virulence is under stabilizing selection across populations was found. Heritability values were high and ranged from 0·52 to 0·96 with a mean heritability of 0·84. Virulence was positively correlated with spore production as predicted by the trade‐off theory of virulence evolution. Furthermore, an association analysis between virulence and sequence haplotypes of three known necrosis‐inducing effector genes (NIP1, NIP2 and NIP3) revealed a significant effect of NIP2 haplotypes and NIP1 deletions. Overall, the results support a quantitative model for virulence in the R. commune–barley pathosystem and very high evolutionary potential for this trait.     

Metabolic and microbial responses in western crossbred and Meishan growing pigs fed a high-fiber diet

Four Duroc x White composite crossbred (21.8 +/- 1.0 kg BW) and four 12-wk-old Meishan purebred (20.7 +/- 1.6 kg BW) growing barrows were used to determine the relative breed differences in metabolic and microbial responses to a high-fiber diet. The pigs were trained to consume 700 g of a diet containing 35% (as-fed basis) dehydrated alfalfa meal once daily. The pigs' daily intakes of DM, N, GE, NDF, and ADF were 610 g, 16.6 g, 2.64 Mcal, 150 g, and 88 g, respectively. On d 12 after surgical catheterization of the portal vein, ileal vein, and carotid artery, a 3-d total urine and feces collection was conducted. On d 24 after surgery, each pig was placed in an open-circuit calorimeter, and its catheters were connected to a system for simultaneous measurements of oxygen consumption by portal-drained viscera and by whole body, and the net portal absorption of VFA after a 24-h fasting and during a 5-h postprandial period. The VFA measured included acetic, propionic, isobutyric, butyric, isovaleric, and valeric acids. A second 3-d total urine and feces collection was conducted on d 30 after surgery. There were no differences (P = 0.13) between the first and second collections in apparent total-tract digestibility coefficients for nutrients and N retention of pigs. Compared with Duroc x White composite crossbred pigs, Meishan pigs had lower (P = 0.05) apparent digestibility coefficients for DM, N, NDF, hemicellulose, and N retention, but their portal-drained viscera used a greater (P = 0.05) fraction of whole-body oxygen consumption. No differences (P = 0.12) were found between Duroc x White composite crossbred and Meishan pigs in total viable bacteria and cellulolytic bacteria from fecal samples, in vitro digestibility of alfalfa NDF fractions by fecal inocula, whole-body oxygen consumption, net portal absorption of VFA, total energy of absorbed VFA, and the potential of absorbed VFA for meeting the energy needs for whole-body heat production. These results indicate that, in contrast to previous beliefs, the ability of Meishan growing pigs to utilize a high-fiber diet is not superior to that of Duroc x White composite crossbred growing pigs.     

Campylobacter monitoring in German broiler flocks: an explorative time series analysis

Campylobacter, a major zoonotic pathogen, displays seasonality in poultry and in humans. In order to identify temporal patterns in the prevalence of thermophilic Campylobacter spp. in a voluntary monitoring programme in broiler flocks in Germany and in the reported human incidence, time series methods were used. The data originated between May 2004 and June 2007. By the use of seasonal decomposition, autocorrelation and cross-correlation functions, it could be shown that an annual seasonality is present. However, the peak month differs between sample submission, prevalence in broilers and human incidence. Strikingly, the peak in human campylobacterioses preceded the peak in broiler prevalence in Lower Saxony rather than occurring after it. Significant cross-correlations between monthly temperature and prevalence in broilers as well as between human incidence, monthly temperature, rainfall and wind-force were identified. The results highlight the necessity to quantify the transmission of Campylobacter from broiler to humans and to include climatic factors in order to gain further insight into the epidemiology of this zoonotic disease.     

Rhabdovirus-like particles in the sporocyst of the trematode Brachylaimus fuscatus from terrestrial mollusc Ponsadenia duplocincta

Rhabdovirus-like particles containing nucleoprotein in a helix with eight turns was demonstrated by transmission electron microscopy in subtegumental cells and syncytial layer of tegument in Brachylaimus fuscatus sporocyst. This is the first case of virus infection in trematodes. The possibility of transfer of the virus to intra-molluscan parasitic generations of the trematode is discussed with respect to the peculiarities of the life cycles of the family Brachylaimidae.     

Inheritance of virulence in Fusarium circinatum,the cause of pitch canker in pines

Wildtype strains of Fusarium circinatum, the causal agent of pitch canker, were crossed to obtain an F₁ generation. Progeny of this cross were tested for virulence by inoculating Pinus radiata seedlings, and were found to induce a wide range of lesion lengths. Two strains from the F₁ generation that induced long lesions (= high virulence) were used as parents to produce an F₂ generation, followed by a second round of selection for high virulence to obtain an F₃ generation. Mean lesion lengths were not significantly different between the three generations (P ≥ 0.196). A parallel set of crosses was performed to select for low virulence by using progeny in the F₁ and F₂ generations that induced short lesions as parents for F₂ and F₃ generations, respectively. In this case, both rounds of selection resulted in a significant reduction in mean lesion length, from 33.8 ± 0.8 mm in the F₁ generation, to 19.7 ± 0.7 and 12.9 ± 0.7 mm in the F₂ and F₃ generations, respectively. Thus it is apparent that F. circinatum retains the genetic capacity for avirulence to pines, which could reflect a lack of strong selection for virulence in nature. Progeny of a cross between high and low virulence parents manifested nearly continuous variation in lesion lengths, consistent with virulence being a quantitatively inherited trait. Based on this cross, broad‐sense heritability (H²) was determined to be 0.74, which suggests that virulence is under strong genetic control.     

Influence of anions on dispersion and physical properties of the A horizon of a Red-brown earth

Three groups of anions were distinguished in order of effectiveness with respect to the dispersion and flocculation of soil samples treated with Fe(III) polycations: phosphate and fulvate citrate, oxalate, silicate and tartrate and salicylate, catechol, aspartate, lactate and acetate. This was also the order of the amount of anion adsorbed by the soil. The addition of phosphate and fulvate to soil samples with a net charge of zero lowered the zero point of charge producing particles with a net negative charge. This increased the amount of dispersible clay present from 0 to 9% by weight of soil. The sorption of phosphate and fulvate by soil samples with a net positive charge reduced the zero point of charge and caused flocculation of dispersed clay. Electrophoretic and electron microscopic studies confirmed the dispersion-flocculation phenomena.Treatments which produced dispersed clay led to increased bulk densities, plastic limits and moduli of rupture but lower porosities, water holding capacities and hydraulic conductivities. The sorption of anions on soil samples with a net charge of zero reduced friability.     

Urethral conformation and position in relation to urinary incontinence in the bitch.

The conformation of the urethra, the position of the bladder neck and the distance between the external urethral orifice and the cranial pubic brim were compared in 30 continent and 30 incontinent bitches with incompetence of the urethral sphincter mechanism, using the measurements made from vagino-urethrograms. The bladder neck was significantly (P less than 0.001) further caudal in incontinent dogs than in continent dogs. Its position was not affected by neutering and could not be explained by the degree of urethral curvature. The distance from the external urethral orifice to the cranial pubic brim was correlated (P less than 0.001) with bodyweight but was not significantly different in the continent and incontinent bitches. In neutered animals, the distance between the cranial pubic brim and the external urethral orifice was significantly (P less than 0.05) shorter than in entire animals.     

Attempted transmission of Ehrlichia risticii by field-captured Dermacentor variabilis (Acari: Ixodidae)

The capability of field-collected American dog ticks, Dermacentor variabilis, to infect horses with Ehrlichia risticii, causative agent of Potomac horse fever (PHF), was examined by allowing adult ticks collected from horse farms with a history of PHF to feed on susceptible horses. More than 500 male and female ticks attached and fed on 3 test horses; however, no clinical or serologic evidence of PHF was observed in treated or control horses. All horses were challenge exposed with E risticii-infective blood by inoculation at 60 to 65 days after ticks fed, and all developed clinical PHF with subsequent seroconversion. The data, therefore, indicated that adult D variabilis, a common parasite of horses on Maryland premises where PHF is enzootic, may not serve as a vector of E risticii.     

Automated identification of sugar beet diseases using smartphones

Cercospora leaf spot (CLS) poses a high economic risk to sugar beet production due to its potential to greatly reduce yield and quality. For successful integrated management of CLS, rapid and accurate identification of the disease is essential. Diagnosis on the basis of typical visual symptoms is often compromised by the inability to differentiate CLS symptoms from similar symptoms caused by other foliar pathogens of varying significance, or from abiotic stress. An automated detection and classification of CLS and other leaf diseases, enabling a reliable basis for decisions in disease control, would be an alternative to visual as well as molecular and serological methods. This paper presents an algorithm based on a RGB‐image database captured with smartphone cameras for the identification of sugar beet leaf diseases. This tool combines image acquisition and segmentation on the smartphone and advanced image data processing on a server, based on texture features using colour, intensity and gradient values. The diseases are classified using a support vector machine with radial basis function kernel. The algorithm is suitable for binary‐class and multi‐class classification approaches, i.e. the separation between diseased and non‐diseased, and the differentiation among leaf diseases and non‐infected tissue. The classification accuracy for the differentiation of CLS, ramularia leaf spot, phoma leaf spot, beet rust and bacterial blight was 82%, better than that of sugar beet experts classifying diseases from images. However, the technology has not been tested by practitioners. This tool can be adapted to other crops and their diseases and may contribute to improved decision‐making in integrated disease control.