A faster inoculation assay for Armillaria using herbaceous plants

Armillaria (honey fungus) is a virulent necrotrophic pathogen that causes Armillaria root disease. Conventional Armillaria inoculation assays use young saplings as hosts and consequently are cumbersome, frequently conducted outdoors, and take many years from establishment to analysis of infection. We have developed and evaluated a faster inoculation assay for Armillaria that uses herbaceous plants as hosts, is carried out in controlled conditions, and reduces experimental durations to 3 months. Plant species of known susceptibility to Armillaria and comparisons between virulent A. mellea and opportunistic A. gallica were used to validate the assay. Mortality and diagnostic symptoms of Armillaria root disease such as epiphytic rhizomorphs and mycelial fans were used to assess levels of infection. We also attempted to reduce assay preparation time by substituting woody inocula with agar inocula, but typical symptoms of Armillaria root disease were only observed on plants infected with woody inocula. Through our assay, we identified five new potential herbaceous hosts of Armillaria: Kniphofia hirsuta, Hordeum vulgare, Lobelia cardinalis, Nicotiana tabacum and Helenium hoopesii – further expanding the extensive list of plants with susceptibility to Armillaria and suggesting infection of herbaceous species may be more widespread than currently acknowledged.     

The Response to Epsom Salt Sprays of Mature Apple Trees of Three Varieties on Two Contrasting Rootstocks

Mature trees of Beauty of Bath, Tydeman’s Late Orange and Laxton’s Superb on M.VII rootstock, growing under commercial-type conditions, were severely deficient in magnesium as shown by leaf analysis and typical symptoms. Five sprays yearly of 2% Epsom salt (MgSO₄.7H₂ O) raised the concentration of magnesium in the leaves and largely eliminated symptoms: the concentrations of calcium and, to a lesser extent, of potassium in leaves were reduced. Over six years, there was little effect of Epsom salt on growth as measured, but crop weight was increased by 19–39% according to variety: this was largely, but not entirely, due to increase in fruit set. The proportion of fruit that dropped was decreased for Beauty of Bath and for Tydeman’s Late Orange sprayed with Epsom salt and the weight per 100 fruits of the late varieties Tydeman’s Late Orange and Laxton’s Superb was increased. Untreated trees on M.II rootstock, in contrast to those on M.VII, showed almost no foliar symptoms of magnesium deficiency, made more growth and produced larger crops; the concentrations of magnesium and calcium in the leaves were higher whereas potassium was usually lower. Epsom salt sprays did not increase growth or cropping; indeed, there was a tendency for sprays to reduce the crop weight of Beauty of Bath on M.II, indicating that they should not be used indiscriminately.

Epsom salt foliar sprays increased the “available” magnesium in the soil to a level considered to be medium—high; this effect was mainly shown in the top 6 in. of soil and gradually diminished to the 18–24 in. zone. Nevertheless, in the year following the termination of treatments, the proportion of foliage affected in Laxton’s Superb on M.VII, previously sprayed with Epsom salt, was almost as great as that of untreated trees and the concentration of magnesium in the leaves was as low. It appeared that magnesium in the soil did not reach the leaves.

The increase in “available” magnesium due to sprays was greater in soil carrying trees on M.II than M.VII, a result probably due to the extra quantity of spray required for these larger trees.     

The kinetics of insecticide action. Part IV: the in-vivo distribution of pyrethroid insecticides during insect poisoning

The distribution of pyrethroids in insects has been studied using a combination of mathematical modelling and experimental observation. This approach has resulted in the formulation of a physiological model of the pharmaco-kinetics of cypermethrin applied topically to larvae of Spodoptera littoralis. Development of this model from simpler two- and three-compartment models is described. Simple models, whilst capable of complex behaviour, consider only the average rates and magnitudes of pharmaco-kinetic processes over whole animals, and cannot account for differences in concentration of insecticide between individual tissues. This can be achieved by using physiological models, but these require more experimental information for their validation. Moreover, unless simplifying assumptions are made, analytical solutions are not feasible for the large number of equations necessary to define such models. The modelling studies prompted an investigation of (1) in-vivo binding of insecticide to insect tissues, (2) the sizes of body compartments, and (3) the factors which affect the distribution of toxicant between these compartments. Binding has a marked effect on pharmacokinetic profiles and may result in oscillatory behaviour. During poisoning, the total bound cypermethrin increases proportionally to the cube root of the elapsed time. This results in a rapid rate of increase over early elapsed times (< 3h) which slows to approach a more linear form thereafter. Average sizes for the body compartments of larvae of Spodoptera littoralis Boisd, and the steady-state distribution of cypermethrin in these compartments are described. Although the haemolymph, which acts as the main distributive phase during poisoning, forms the largest compartment by volume, it has a low affinity for cypermethrin and distribution reaches steady state within 5 min after topical application. The nerve cord (the target tissue), which is the smallest compartment, has the highest steady-state concentration of cypermethrin. The distribution of cypermethrin in larval tissues is related to the ratios of tissue dry matter to water content.     

The role of altered uterine-embryo synchrony on conceptus growth in the pig

This study was conducted to determine whether inducing an embryo-uterine asynchrony during the preimplantation period would alter fetal and(or) placental size at term. Yorkshire gilts (n = 24) were checked twice daily for estrus and bred to a Yorkshire boar 24 h after the first exhibition of estrus. Embryos (1 to 4 cells) were flushed from the oviducts of each donor gilt on d 2.5 of gestation and transferred in equal numbers to the oviducts of a recipient gilt on d 1.5, 2.5, or 3.5 of the estrous cycle. Gilts were slaughtered on d 112 of gestation (calculated on the age of the conceptus) and fetal and placental weight, placental surface area, and implantation site lengths were determined. Although litter sizes were similar (9.1+/-0.9), conceptuses transferred to d 3.5 recipients became heavier fetuses (1.44+/-0.05 vs 1.23+/-0.04 kg, P < 0.001), with larger placental surface areas (1,793+/-60 vs 1,459+/-43 cm2, P < 0.01), and longer implantation sites (32.1+/-1.5 vs 24.9+/-0.6 cm, P < 0.001) than those transferred to recipients on d 2.5. These data demonstrate that oviductal transfer of embryos into a reproductive tract that is more advanced by as little as 24 h can result in alterations in placental growth and function during gestation.     

Pleuroscopic diagnosis of gastroesophageal squamous cell carcinoma in a horse

A 15-year-old Quarter Horse gelding was examined because of chronic intermittent colic of 40 days' duration. The clinical signs included acute onset of depression, ptyalism, abdominal splinting, and rolling within an hour of eating. An intramural mass of the esophagus was identified during esophagoscopy. A large soft-tissue density surrounding the distal portion of the esophagus, creating a stricture, was identified on an esophagogram. With the horse slightly sedated, pleuroscopy was performed, allowing direct visualization and biopsy of the mass. The histopathologic diagnosis was squamous cell carcinoma.     

Relationships of testicular iron and ferritin concentrations with testicular weight and sperm production in boars

The inverse relationship of testicular size and circulating follicle-stimulating hormone (FSH) concentrations has been documented, and accompanying this relationship is the change in color of the parenchymal tissue of the testes. Large testes (300 to 400 g) are pink to light red and small testes (100 g) are dark maroon with color gradations for weights in between. It was hypothesized that this color most likely represented an iron protein. Chromatographic analysis of testicular tissue indicated that the Fe was associated primarily with ferritin, and immunohistochemistry showed that Leydig cells were the primary location of ferritin storage within the testes. Concentrations of Fe and ferritin were higher in small testes and decreased as testes weight increased (P < 0.05). As testicular Fe concentrations increased, daily sperm production (DSP) and total DSP declined (P < 0.05). Genotyping six generations of Meishan x White composite boars (n = 288) for a quantitative trait locus that is indicative of elevated FSH and small testes in boars indicated that the Meishan genotype had elevated testicular iron concentrations and darker color in conjunction with reduced total DSP (P < 0.01). It is not thought the elevated iron concentrations affect testicular weights but are probably a result of elevated FSH and FSH inducement of Fe transport. The storage of Fe in Leydig cells may provide a reservoir of Fe for easy access by Sertoli and germ cells, but still provide a degree of protection to germ cells from ionic iron.     

GnRH antagonist inhibition of gonadotropin and steroid secretion in boars in vivo and steroid production in vitro

The hormone GnRH has a stimulatory effect on gonadotropin synthesis and secretion. The objective of the first study was to evaluate concentrations of FSH and LH in plasma of boars after successive treatment with SB75, a GnRH antagonist. Thirteen boars greater than 1 yr of age (eight White Composite [WC] and five Meishan [MS]) were injected once daily with SB75 (10 microg/kg of body weight) for 4 d. Plasma concentrations of LH and testosterone (T) decreased after 1 h from the first dose of SB75. After 12 h of treatment, LH gradually returned to pretreatment concentrations, but T remained suppressed (< 2 ng/mL) until after the last injection of SB75. There was a modest, but significant, reduction in FSH during treatment with SB75. The prolonged inhibitory effect of SB75 on suppression of plasma T concentrations, in the presence of pretreatment concentrations of LH, implied direct effects of SB75 at the testis. In the second experiment, testicular tissue from adult boars was incubated in the presence of three doses of human chorionic gonadotropin (hCG; 0, .5, and 5 IU) with SB75 (250 ng/mL) or with Deslorelin, a GnRH agonist (500 ng/mL). Samples of media were collected every hour for 3 h, and concentrations of T and estrone (E1) were determined by RIA. Concentrations of T and E1 increased with time in response to treatment with hCG. Co-treatment with SB75 decreased media concentrations of T (P < .01) and E1 (P < .03) compared to controls (77.9 vs 85.7 +/- 2.0 and 4.7 vs 5.3 +/- .2 ng/g). In contrast, treatment with Deslorelin had no effect on the amount of T (P > .50) or E1 (P > .26) released with all dosages of hCG. These results indicate that a GnRH antagonist has a direct effect on the testis, decreasing amounts of T and E1 released from the Leydig cells; however, treatment with a GnRH agonist had no direct effect on release of these gonadal steroids. Thus, it remains unresolved whether the site of action of GnRH antagonist on testicular steroidogenesis is through a testicular GnRH receptor or through some other mechanism.     

Hypercalcemia in a dog: a challenging case

An 18-month-old, spayed female, mixed-breed dog was referred for investigation of persistent hypercalcemia. After extensive diagnostic evaluation, a tentative diagnosis of occult lymphosarcoma (LSA) was made and the dog was euthanized. At necropsy, infection with Heterobilharzia americana was diagnosed. In endemic areas, schistosomiasis should be included in the differential diagnosis of hypercalcemia, and a fecal examination should be performed in every dog with a hypercalcemia of unknown origin.     

Bluetongue virus serotypes 1 and 3 infection in Poll Dorset sheep

Objective To study the clinical signs following bluetongue virus serotypes 1 and 3 infection in Poll Dorset sheep.
Design A clinical and pathological study.
Procedure Twenty Poll Dorset sheep were inoculated with bluetongue virus serotypes 1 or 3, each inoculum having a different passage history. The sheep were examined daily and their clinical appearance and rectal temperatures recorded. Heparinised and non-heparinised blood samples were taken at intervals for virological and serological study. Gross pathological findings were recorded for several sheep at necropsy and tissue samples were collected from three sheep for virological studies.
Results All inoculated sheep developed clinical disease. The clinical signs and gross pathological changes varied considerably but were consistent with damage to the vascular endothelial system. There was a decline in the titres of infectious bluetongue virus and of antigen in tissues collected between 7 and 12 days after infection.
Conclusions The severity of disease was related to the speed of onset and duration of pyrexia and not the development or titre of viraemia. Generally, those animals with sensitive mouths, depression, coronitis, recumbency and reluctance to move were the most debilitated. Whole blood was the most reliable source of infectious virus from acutely and chronically infected and convalescent animals. However, tissue samples particularly spleen, collected from dead or killed animals suffering from either peracute or acute forms of disease were most appropriate for the rapid confirmation of a clinical diagnosis.