A duplex-PCR bioassay to detect a Trichoderma virens biocontrol isolate in non-sterile soil

This paper describes the identification and utilisation of a sequence-characterised amplified region (SCAR) marker specific for the Trichoderma virens biocontrol isolate GV4. The marker was developed from a RAPD-PCR amplification product unique to isolate GV4. When used as a hybridisation probe in Southern blot analysis, it hybridised to the DNA of the species T. virens alone and not to that of other Trichoderma species or closely related genera Gliocladium and Verticillium. The marker also produced a GV4-specific RFLP, distinguishing it from other T. virens isolates when probed to blots with HindII, BamHI or PstI genomic DNA digests. Primers designed from the sequence of the RAPD marker produced a diagnostic amplification product of 346 bp for GV4 alone, distinguishing it from all other test isolates. With the exception of one, test isolates did not produce an amplification product with the SCAR primers. The exception was a single Verticillium psalliotae isolate (ICMP5509) that produced a product of approx. 400 bp that was easily distinguished from the 346 bp product of GV4. The reliability of the SCAR-based diagnostic test was further improved with the introduction of a positive PCR reaction control to each test, achieved by converting the test to a duplex PCR system. Two universal primers flanking the two ITS and the 5.8S region of the ribosomal gene complex were introduced to each reaction to provide a test for PCR reaction inhibitors to eliminate false negatives in the diagnosis. Amplification of this multi-copy genomic region did not reduce diagnostic sensitivity of the single copy SCAR marker. To further increase the sensitivity of detecting GV4 propagules while maintaining a fast sample assessment assay, soil was amended with cornmeal, as a nutrient source, and a mix of antibiotics to favour Trichoderma growth. The soil mix was subsequently incubated for 5 d before total DNA was extracted. Under these conditions, the duplex soil PCR assay detected GV4 down to a concentration of 10 spores g⁻¹ soil in non-sterile agricultural field soil. This study is the first to report the use of a duplex-PCR diagnostic bioassay for a species within the Hypocrea/Trichoderma genus.     

MULTICENTRIC LYMPHOMA WITH INTESTINAL INVOLVEMENT IN A DOG

A 5-year-old, male German shepherd dog was evaluated for a 5-week history of vomiting, diarrhea, lethargy, and weight loss. In abdominal radiographs, there was an unusual corrugated appearance within a loop of intestine. Sonographically, this loop also had a similar corrugated appearance with alternating hyperechoic and hypoechoic bands. Lymph nodes within the abdomen and thorax were enlarged, and a thoracic mass was also detected. Ultrasound-guided fine-needle aspirates of the mesenteric lymph nodes were highly suggestive of lymphoma. After postmortem examination, a diagnosis of multicentric lymphoma with involvement of the gastrointestinal tract was made.     

Morphology and classification of 96 primary cardiac neoplasms in Fischer 344 rats

Ninety-six primary cardiac neoplasms were identified from 79,971 Fischer 344 (F344) rats used in chronic toxicity and carcinogenicity studies by the National Toxicology Program (NTP) and National Cancer Institute (NCI), for an overall incidence of 0.1%. Neoplasms were classified as: 60 endocardial schwannomas, 23 intramural schwannomas, eight atriocaval mesotheliomas, three paragangliomas, one pericardial mesothelioma, and one hemangioma. Metastases occurred in four rats with endocardial schwannoma. Histological appearance of the endocardial and intramural schwannomas was consistent with origin from nerve sheath. Two of six endocardial schwannomas available for immunohistochemical staining were weakly positive for S-100 antigen. The atriocaval mesotheliomas, while morphologically resembling adenocarcinoma, were positive for vimentin and keratin, indicating mesothelial origin. Seventy of the 96 cardiac neoplasms occurred in rats 2 years of age or older at time of death. There were no sex or treatment-related differences in the incidence of these neoplasms, with the exception of atriocaval mesothelioma, which was more common in males.     

Ovarian choriocarcinoma in the mouse

Choriocarcinoma is one of the rarest ovarian tumors in any animal species. This paper describes the gross and microscopic appearance of seven such neoplasms in B6C3F1 mice. Mean age at death was 47 weeks. Tumors were described at necropsy as dark or hemorrhagic cystic lesions. On microscopic examination tumors were composed of hematocysts, intercellular hemorrhage, and cytotrophoblasts, syncytiotrophoblasts, and/or trophoblastic giant cells. Cytotrophoblasts, syncytiotrophoblasts, and occasional giant cells were present in three cases while the other four tumors contained only trophoblastic giant cells.     

Mechanisms of cuticular uptake of xenobiotics into living plants: 1. Influence of xenobiotic dose on the uptake of three model compounds applied in the absence and presence of surfactants into Chenopodium album, Hedera helix and Stephanotis floribunda leaves

This study determined the uptake of three model compounds, applied in the presence and absence of surfactants, into the leaves of three plant species (Chenopodium album L, Hedera helix L and Stephanotis floribunda Brongn). The results with 2-deoxy-D-glucose, 2,4-dichlorophenoxyacetic acid and epoxiconazole in the presence ofsurfactants (the polyethylene glycol monododecyl ethers C12EO3, C12EO6, C12EO10 and a trisiloxane ethoxylate with mean EO of 7.5, all used at one equimolar concentration and therefore different percentage concentrations) illustrate that the initial dose (nmol mm(-2)) of xenobiotic applied to plant foliage is a strong positive determinant of uptake. This held true for all the xenobiotics studied over a wide concentration range in the presence of these surfactants. Uptake on a unit area basis (nmol mm(-2)) could be related to the initial dose of xenobiotic applied per unit area (ID) by an equation of the form: Uptake = a [ID]b at time t = 24h. ID is given by the mass of xenobiotic applied, M divided by the droplet spread area, A. Total mass uptake is then calculated from an equation of the form: Total Uptake = a [ID]b x A.     

Liverpool Veterinary School: the first 100 years

In 1904, Liverpool veterinary school became the first UK veterinary school to be incorporated into a university. This weekend, it marks its centenary with a symposium celebrating veterinary science and its contribution to animal and human health and welfare. Here, Alison Kraft traces the history of the school and argues that its foundation was a turning point for veterinary education in the UK.     

A biomechanical comparison of headless tapered variable pitch compression and ao cortical bone screws for fixation of a simulated midbody transverse fracture of the proximal sesamoid bone in horses

OBJECTIVE: To compare mechanical properties and failure characteristics of 2 methods of fixation for repair of a transverse, midbody fracture of the proximal sesamoid bone (PSB): 4.5-mm AO cortical bone screw (AO) placed in lag fashion and 4/5-mm Acutrak (AT) self-compressing screw. STUDY DESIGN: An in vitro biomechanical evaluation of intact forelimb preparations and forelimb preparations with a simulated midbody PSB fracture stabilized by a bone screw. SAMPLE POPULATION: Sixteen paired and 8 unilateral cadaveric equine forelimbs. METHODS: A midbody transverse osteotomy was created in the medial PSB of bilateral forelimbs of 8 equine cadavers. The osteotomized PSB in 1 forelimb from each cadaver was repaired with an AO screw. The osteotomized PSB in each contralateral limb was repaired with an AT screw. Eight unilateral intact control limbs were also studied. Mechanical properties were determined from axial compression, single cycle to failure, load-deformation curves. Failure characteristics were determined by evaluation of video images and radiographs. RESULTS: No statistically significant differences were found between repair groups. Both AO and AT groups had significantly lower mechanical properties than intact limbs except for stiffness. CONCLUSION: AO and AT constructs were mechanically comparable when used to stabilize a simulated midbody fracture of the medial PSB. Both constructs were mechanically inferior to intact limbs. Clinical Relevance- The AT screw should be considered for clinical use because of the potential for less soft tissue impingement and superior biocompatibility compared with the stainless-steel AO screw. However, postoperative external coaptation is necessary to augment initial fracture stability for either fixation method, and to maintain a standing metacarpophalangeal joint dorsiflexion angle between 150 degrees and 155 degrees.     

Mechanisms of cuticular uptake of xenobiotics into living plants: evaluation of a logistic-kinetic penetration model

The objective of this study was to determine whether a logistic-kinetic penetration model could be applied to whole plant uptake. Uptake over 24 h was determined for three model compounds, applied in the presence and absence of surfactants, into the leaves of two plant species. Data for two time intervals were used in the model to predict uptake at intermediate intervals and compared with experimental results. Overall, the model fit the whole plant uptake data well. The study confirmed that an increase (or decrease) in active ingredient (ai) concentration or an increase in contact area will have no effect on the penetration rate factor, q, within the normal working concentration range. This enabled uptake to be predicted at different times for concentrations of ai not already studied, having first derived q for one concentration of the formulation of interest and having 24 h (maximum) uptake results for all formulations and concentrations of interest. The advantages of the models and equations described are that few variables are required, and they are simple to measure.