A quantitative real‐time PCR assay for detection of Colletotrichum lindemuthianum in navy bean seeds

Bean anthracnose is a seedborne disease of common bean (Phaseolus vulgaris) caused by the fungal pathogen Colletotrichum lindemuthianum. Using seed that did not test positive for the pathogen has been proven to be an effective strategy for bean anthracnose control. To quantify the extent of anthracnose seed infection, a real‐time PCR‐based diagnostic assay was developed for detecting C. lindemuthianum in seeds of the commercial bean class navy bean. The ribosomal DNA (rDNA) region consisting of part of the18S rDNA, 5^.8S rDNA, internal transcribed spacers (ITS) 1, 2 and part of the 28S rDNA of seven races of C. lindemuthianum, 21 isolates of Colletotrichum species and nine other bean pathogens were sequenced with the universal primer set ITS5/ITS4. Based on the aligned sequence matrix, one primer set and a probe were designed for a SYBR Green dye assay and a TaqMan MGB (minor groove binder) assay. The primer set was demonstrated to be specific for C. lindemuthianum and showed a high sensitivity for the target pathogen. The detection limit of both assays was 5 fg of C. lindemuthianum genomic DNA. To explore the correlation between the lesion area and the DNA amount of C. lindemuthianum in bean seed, seeds of the navy bean cultivar Navigator with lesions of different sizes, as well as symptomless seeds, were used in both real‐time PCR assays.     

Nutrient availability and corn growth in a poultry litter biochar‐amended loam soil in a greenhouse experiment

Nutrient‐rich biochar produced from animal wastes, such as poultry litter, may increase plant growth and nutrient uptake although the role of direct and indirect mechanisms, such as stimulation of the activity of mycorrhizal fungi and plant infection, remains unclear. The effects of poultry litter biochar in combination with fertilizer on mycorrhizal infection, soil nutrient availability and corn (Zea mays L.) growth were investigated by growing corn in a loam soil in a greenhouse with biochar (0, 5 and 10 Mg/ha) and nitrogen (N) and phosphorus (P) fertilizer (0, half and full rates). Biochar did not affect microbial biomass C or N, mycorrhizal infection, or alkaline phosphomonoesterase activities, but acid phosphomonoesterase activities, water‐soluble P, Mehlich‐3 Mg, plant height, aboveground and root biomass, and root diameter were greater with 10 Mg/ha than with no biochar. Root length, volume, root tips and surface area were greatest in the fully fertilized soil receiving 10 Mg/ha biochar compared to all other treatments. The 10 Mg/ha biochar application may have improved plant access to soil nutrients by promoting plant growth and root structural features, rather than by enhancing mycorrhizal infection rates.     

Establishment and dissemination of hypovirulent strains of Cryphonectria parasitica in Greece

Chestnut blight caused by Cryphonectria parasitica has been one of the major reasons for the decline of chestnut cultivation in Greece over the last 50 years. A previous detailed study of the vc types of the fungus has revealed only four vc types in the entire country, those of EU‐1, EU‐2 and EU‐10 with the dominant being EU‐12 counting for 88% of the isolates. As the loss in orchard trees reached in some chestnut producing prefectures over 35% of the trees, the decision was taken to apply biological control on a nationwide scale. The project was implemented in 17 prefectures during the period 2007–2009. Hypovirulent (hv) strains of C. parasitica which were infected by the CHV‐1 subtype I (Italian subtype) viruses which occur naturally in Greece were used. The necessary hypovirulent inoculum was prepared at the Forest Research Institute during the period 2007–2009 on a large scale, however, with laboratory care. Three million inoculations were made around accessible developing cankers over three consecutive years by trained personnel. An extensive evaluation conducted in 2011 in the 12 of the 17 treated prefectures, where no natural hypovirulence had been found before, demonstrated not only the successful establishment of hypovirulence but also profound dissemination and healing of non‐inoculated cankers. The extent of dissemination varied significantly among sampling plots and among prefectures. The conclusion is drawn that CHV‐1 subtype I has successfully established and spread into chestnut orchards and coppice forests resulting in a gradual decline of chestnut blight.     

Transmission characteristics of low pathogenic avian influenza virus of H7N7 and H5N7 subtypes in layer chickens

Low pathogenic avian influenza virus (LPAIv) infections of H5 and H7 subtypes in poultry are notifiable to the OIE, hence surveillance programmes are implemented. The rate at which LPAIv strains spread within a flock determines the prevalence of infected birds and the time it takes to reach that prevalence and, consequently, optimal sample size and sampling frequency. The aim of this study was to investigate the transmission characteristics of an H7N7 and an H5N7 LPAIv in layer chickens. Two transmission experiments were performed, which consisted of 30 (first experiment) and 20 (second experiment) pairs of conventional layers, respectively. At the start of the experiments, one chicken per pair was inoculated with LPAIv and the other chicken was contact-exposed. Occurrence of infection was monitored by regularly collecting tracheal and cloacal swab samples, which were examined for the presence of virus RNA by RT-PCR. The results of the test were used to estimate the transmission rate parameter (β), the infectious period (T) and the basic reproduction ratio (R(0)). In addition, egg production and virus shedding patterns were quantified. For the H7N7 virus, the β, T and R(0) estimates were 0.10 (95% confidence interval (CI): 0.04-0.18) day(-1), 7.1 (95% CI: 6.5-7.8) days and 0.7 (95% CI: 0.0-1.7), respectively. With the H5N7 virus, only a few inoculated chickens (5 out of 20) became infected and no transmission was observed. This study shows that transmission characteristics of LPAIv strains may vary considerably, which has to be taken into account when designing surveillance programmes.     

The pathology of spontaneous paratuberculosis in the North American bison (Bison bison)

Gross and histopathologic examinations were performed on 70 North American bison (Bison bison) from a Mycobacterium avium paratuberculosis culture-positive herd. The bison examined were part of a breeding herd totaling 2,800 animals. Eight of 70 (11%) animals had gross findings of intestinal mucosal thickening, and 16 of 70 (23%) of the animals had enlarged mesenteric lymph nodes. Histologic lesions compatible with Johne's disease were diagnosed in 30 of 70 (43%) bison on the basis of the demonstration of noncaseating granulomatous inflammatory infiltrates and of one or more acid-fast bacilli characteristic of Mycobacterium avium paratuberculosis. A suspicious diagnosis of Johne's disease was obtained in 11 of 70 (16%) bison on the basis of the observation of noncaseating granulomatous inflammatory infiltrates without demonstrable acid-fast bacteria. Twenty-nine of 70 (41%) animals were assessed as histologically paratuberculosis free. Histologic results were compared to Johne's disease tests such as culture, serology, and polymerase chain reaction, which were performed on some of the cohort animals.     

Milk and serum concentration of ceftiofur following intramammary infusion in goats

Five dairy goats were used to determine the milk and serum concentrations along with elimination characteristics of ceftiofur following intramammary administration. One udder half of each goat was infused twice with 125 mg ceftiofur with a 24‐h interval between infusions. Milk samples were collected at 1, 2, 8, and 12 h after the last infusion and then every 12 h for a total of 7 days. Blood was collected from each animal at 3, 8, 12, and 24 h after infusion and then every 24 h for 6 days. Following a washout period of 1 week, the experiment was repeated using the opposite udder half. The elimination half‐life of ceftiofur from the mammary gland was 4.7 h. The concentration of ceftiofur was greater than published MIC₉₀ values for Staphylococcus spp. bacteria for 24 h. Ceftiofur was absorbed into systemic circulation from the mammary gland. The maximum concentration was 552 ng/mL at 3 h after infusion, and the serum elimination half‐life was 10 h. Intramammary infusion of 125 mg ceftiofur every 24 h can be expected to maintain drug concentration in milk above published MIC₉₀ for Staphylococcus spp.     

Sediment production from unpaved roads in a sub-tropical dry setting — Southwestern Puerto Rico

The threat imposed by increased sediment loading rates ranks among the most important stressors affecting coral reef ecosystems worldwide. This study represents an effort to quantify the effects of unpaved roads on erosion rates in a dry sub-tropical area of Puerto Rico and is intended to aid in developing scientifically-based erosion mitigation strategies. Hence, the specific objectives of this study were to: (1) measure sediment production rates from unpaved roads; (2) evaluate the effect of precipitation, rainfall erosivity, slope, plot length, and vegetation cover on sediment production rates; and (3) compare measured sediment production rates to published surface erosion data from roaded and natural sites in the Eastern Caribbean. Sediment production from nine abandoned road segments with varying slopes and plot lengths were measured with sediment traps in southwestern Puerto Rico from August 2003 to September 2005.     

Biplot analysis of trait relations of white lupin in Spain

A 2-year study of autumn-sown white lupin (Lupinus albus L.) was conducted under rainfed Mediterranean conditions in southern Spain. 108 landraces were evaluated for yield and major phonological and plant structural characteristics. Path coefficient and Genotype-Trait (GT) biplot analysis were used. Applying both types of analyses to the multiple trait data revealed that GT biplot graphically displayed the interrelationships among traits and facilitated visual comparison of accessions and selection. Wide variation was observed for trait phenology (dates of flowering and maturity), plant structure (plant and first order inflorescence height) and yield characters (first order branches, first order inflorescence and total yield). It was found that the accessions with the highest yields were the tallest and the latest to flower, both on the mainstem and on the lowest first order branch, and also were the latest to mature. GT biplots also had suggested as the accessions have been displayed according to their origins. The results showed that foreign germplasm may be an important material for improving some yield components such as number of pods and seeds per plant of locally adapted lupins in Spain. This revised version was published online in July 2006 with corrections to the Cover Date.     

THE USE OF ELECTROCARDIOGRAPHY IN THE DIAGNOSIS OF POOR RACING PERFORMANCE IN THE HORSE

Electrocardiograms were reocrded from 153 racehorses and 10 Olympic three-day-event horses. The horses were then grouped into those presented for routine examination, those in which there was a complaint of poor performance, those going to the Montreal Olympics, and those with upper respiratory tract abnormalities. When the ECGs of the four groups were compared on the basis of detection of abnormality, it was found that 93.2% of the poor performance group showed abnormalities of various types. The routine examination group showed 21.1% of horses with abnormal ECGs, and over half of these have been followed to the point where it can be said they raced unsuccessfully. In particular, this applied to those horses with T wave abnormalities in 4 or more leads, and to those with intraatrial block. There were significant T wave changes in 50% of the Olympic horses, and 43.7% of horses with laryngeal hemiplegia showed abnormality. While it can be concluded that T wave changes are highly significant findings in an ECG, it is also true that modification of the training programme is a way of helping trainers to get the best out of horses which might otherwise have a serious limitation of their performance potential.