Development of an Arabidopsis thaliana-based bioassay for investigating seed colonization by mycotoxigenic Aspergillus species

Seeds of Arabidopsis thaliana are hosts for three mycotoxin-producing Aspergillus species, A. flavus , A. nidulans and A. parasiticus, enabling an Aspergillus - Arabidopsis infection (AAI) assay to be developed. The AAI assay involved inoculation of 10- to 12-mg aliquots of uniformly cultivated, surface-sterilized A. thaliana seeds in microcentrifuge tubes. Use of microcentrifuge tubes facilitated qualitative and quantitative analyses of post-infection characteristics such as sporulation and mycotoxin production. Cultivation of A. thaliana seeds under uniform environmental conditions is necessary to limit genotype-independent seed-lot variability. Using the A. nidulans oxylipin mutant, Δ ppoABC , and two well-characterized A. thaliana pathogen-defence mutants, ein2-1 and pad4-1 , the AAI assay permitted genetic analysis of seed infection and mycotoxin production. Sporulation, but not mycotoxin production, was impaired in A. nidulans ΔppoABC , while A. thaliana ein2-1 and pad4-1 had a small but detectable influence on A. nidulans sporulation that appeared to be dependent on seed age.     

Biological and molecular characterization of a novel carmovirus isolated from angelonia

ABSTRACT A new carmovirus was isolated from Angelonia plants (Angelonia angustifolia), with flower break and mild foliar symptoms, grown in the United States and Israel. The virus, for which the name Angelonia flower break virus (AnFBV) is proposed, has isometric particles, approximately 30 nm in diameter. The experimental host range was limited to Nicotiana species, Schizanthus pinnatus, Myosotis sylvatica, Phlox drummondii, and Digitalis purpurea. Virions were isolated from systemically infected N. benthamiana leaves, and directly from naturally infected Angelonia leaves, using typical carmovirus protocols. Koch's postulates were completed by mechanical inoculation of uninfected Angelonia seedlings with purified virions. Isometric particles were observed in leaf dips and virion preparations from both Angelonia and N. benthamiana, and in thin sections of Angelonia flower tissue by electron microscopy. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis of dissociated purified virus preparations, a major protein component with a molecular mass of 38 kDa was observed. Virion preparations were used to produce virus-specific polyclonal antisera in both Israel and the United States. The antisera did not react with Pelargonium flower break virus (PFBV), Carnation mottle virus (CarMV), or Saguaro cactus virus (SgCV) by either enzyme-linked immunosorbent assay or immunoblotting. In reciprocal tests, antisera against PFBV, CarMV, and SgCV reacted only with the homologous viruses. The complete nucleotide sequence of a Florida isolate of AnFBV and the coat protein (CP) gene sequences of Israeli and Maryland isolates were determined. The genomic RNA is 3,964 nucleotides and contains four open reading frames arranged in a manner typical of carmoviruses. The AnFBV CP is most closely related to PFBV, whereas the AnFBV replicase is most closely related to PFBV, CarMV, and SgCV. Particle morphology, serological properties, genome organization, and phylogenetic analysis are all consistent with assignment of AnFBV to the genus Carmovirus.     

RESEARCH PAPER: Incidence of elevation of cardiac troponin I prior to and following routine general anaesthesia in dogs

ObjectiveTo estimate the incidence of raised cTnI after general anaesthesia in dogs and to explore major risk factors influencing this.Study designProspective clinical study.AnimalsA total of 107 (ASA physical status 1?2) dogs, 63% male and 37% female, median age 5 years (range 0.3–13.4), median weight 24.4 kg (range 4.2–66.5 kg) undergoing anaesthesia for clinical purposes.MethodsVenous blood samples were taken within 24 hours prior to induction and 24 hours after the termination of anaesthesia. Serum concentrations of cardiac troponin I were measured using a chemiluminescent enzyme immunometric assay with a lower level of detection of 0.20 ng mL^?1 (below this level <0.20 ng mL^?1). Continuous data were assessed graphically for normality and paired and unpaired data compared with the Wilcoxon signed ranks and Mann–Whitney U‐tests respectively. Categorical data were compared with the Chi squared or Fisher’s exact test as appropriate (p < 0.05).ResultsOf the 107 dogs recruited, 100 had pre‐ and post‐anaesthetic cTnI measured. The median pre‐anaesthesia cTnI was ‘<0.20’ ng mL^?1 (range ‘<0.20’–0.43 ng mL^?1) and the median increase from pre‐anaesthesia level was 0.00 ng mL^?1 (range ?0.12 to 0.61 ng mL^?1). Fourteen dogs had increased cTnI after anaesthesia relative to pre‐anaesthesia (14%, 95% CI 7.2–20.8%, range of increase 0.03–0.61 ng mL^?1). Six animals had cTnI levels that decreased (range 0.02–0.12 ng mL^?1). Older dogs were more likely to have increased cTnI prior to anaesthesia (OR = 5.32, 95% CI 1.35–21.0, p = 0.007) and dogs 8 years and over were 3.6 times as likely to have an increased cTnI after anaesthesia (95% CI 1.1–12.4, p = 0.028).Conclusion and clinical relevanceIncreased cTnI after anaesthesia relative to pre‐anaesthesia levels was observed in a number of apparently healthy dogs undergoing routine anaesthesia.     

Reproductive and growth responses of gilts to exogenous porcine pituitary growth hormone

Forty gilts (mean wt = 72 kg) were administered daily either vehicle (C = control) or 70 micrograms porcine growth hormone (pGH)/kg BW. After 30 d of treatment, eight gilts per group (Exp. 1) were slaughtered and blood, uteri and ovaries were collected. Follicular fluid (FFl) was collected and granulosa cells (GC) were cultured. The remaining gilts (Exp. 2) were treated for up to 35 additional days and examined twice daily for estrus. Estrusal gilts were removed from the experiment. Noncyclic gilts (n = 9 of 12 pGH; n = 4 of 12 C) were slaughtered on d 66 and their ovaries were examined. Ovarian weights were not different for pGH and C gilts in either Exp. 1 (P greater than .1) or Exp. 2 (P = .09). Uterine weights were greater for pGH-treated than for C gilts (P less than .007) in Exp. 1, but not in Exp. 2. Concentrations of estradiol (E2) in plasma and FF1 and of progesterone (P) in plasma and FF1 were not different for pGH and C gilts. Concentrations of insulin-like growth factor-I (IGF-I) in FF1 and in serum were greater for pGH than for C gilts (P less than .01). Concentration of P in serum-free medium of cultured GC was lower for GH than for C (P less than .05) in the presence or absence of gonadotropins in Exp. 1. The FSH-stimulated secretion of P was also lower for GC of pGH-treated gilts in Exp. 2, indicating a failure of GC to differentiate in culture. Only one pGH gilts in Exp. 2 manifested estrus, compared with seven C gilts (P less than .025). In Exp. 1, ADG was higher (P less than .03) and feed/gain lower (P less than .07) for pGH gilts. Longissimus muscle area (LMA) was not different (P = .19) between groups. Backfat thickness (BF) was lower (P less than .005) in pGH than in C in both Exp. 1 and 2. We conclude that exogenous pGH increased growth rate, improved feed efficiency and altered carcass traits in gilts. However, these effects were associated with impaired ovarian development of prepubertal gilts and a low incidence of estrus.     

Additive and nonadditive differences in postweaning growth and carcass characteristics of Devon, Hereford, and reciprocal-cross steers.

Postweaning growth and carcass characters of 110 steers from a complete two-breed diallel of the Devon and Hereford breeds were examined under two environments. Additive and nonadditive effects were estimated using linear contrasts for several growth and carcass traits. Steers from each of the four breed groups were grown postweaning to slaughter in high- and low-nutrition environments. Weights were recorded every 2 mo. At slaughter, hot carcass weight, longissimus muscle area, kidney and channel fat, and subcutaneous fat at nine sites were measured. Heterosis for postweaning growth rate was 3.9% (P less than .01) and for slaughter weight 5.0% (P less than .01). Within the low-nutrition environment during periods of slow and fast growth, the Devons and Herefords performed differently. The growth rate of the steers differed in the two environments; however, heterosis for slaughter weight was of the same magnitude in both environments. No differences existed between the straightbreds or between the reciprocal crosses for slaughter weight. Crossbred carcasses were 7.4% heavier (P less than .01) than the straightbred carcasses; however, this effect was removed after adjustment for differences in slaughter weight. Heterosis for longissimus muscle area and carcass fatness were not significant after adjusting for carcass weight. Additive differences occurred for carcass traits. Devon carcasses had more kidney and channel fat (P less than .05) at a constant hot carcass weight and differences occurred in the partitioning of fat within the subcutaneous depot. No significant maternal effects were observed for the carcass traits measured. Crossbreeding increased carcass weight without altering composition, and relative performance was not affected by the diverse environments.     

Live animal measurement of carcass traits by ultrasound: assessment and accuracy of sonographers.

The establishment and evaluation of an assessment system to accredit sonographers for measuring the carcass traits of subcutaneous fat depths and longissimus muscle area (LMA) on potential breeding animals by real-time ultrasound is described. Repeatability of operators, variation between the animal's left and right sides, and variations in technique were assessed from measurements and repeat measurements of 30 cattle by up to eight operators at three testing sessions. Accuracy of carcass data was determined by repeatability of measurements, variability between measurers, between left and right sides of the carcass, and variation due to handling and dressing procedures. Correlations with carcass data averaged .92 for rump fat, .90 for rib fat, and .87 for LMA. Residual SD averaged .81 mm, .88 mm, and 5.1 cm2. A very experienced sonographer can measure LMA only marginally less accurately than it can be measured on the carcass. In Session 3, the SE between repeat fat measurements for accredited sonographers averaged .43 mm, indicating that fat depths can be measured more accurately, but when comparing measurements from different operators, adjustments may be required for differences in technique, otherwise overall accuracy will be about the same, approximately 1 mm. Scanned rump fat measurements were consistently approximately 20% higher than on the chilled, hanging carcass 24 h after slaughter; after applying the standard correction factor of 1.17, LMA measurements were similar. Scan and carcass rib fat measurements were similar for animals with less than or equal to 10 mm of fat cover, above which carcass measurements tended to be higher.