Application of a putative fatty-acid binding protein to discriminate serologically the two European quarantine root-knot nematodes, Meloidogyne chitwoodi and M. fallax, from other Meloidogyne species

Two major proteins, Mcf-A67 and Mcf-B66, were identified by mini two-dimensional polyacrylamide gel electrophoresis in order to distinguish the two European quarantine root-knot nematodes, Meloidogyne chitwoodi and M. fallax, from eight other species. These quarantine proteinic markers have been microsequenced after enzymatic digestion. The internal amino acid sequences exhibit similarities to members of a family of low molecular weight intracellular lipid-binding proteins. Moreover, to explore a simple, rapid, and inexpensive way to identify the two quarantine nematodes, dot blot hybridizations were performed using an antiserum (A67) produced from the longest amino-acid sequence of the protein Mcf-A67. Although several proteins stained on the M. chitwoodi and M. fallax western blot membranes, the two nematodes were easily distinguished from other root-knot nematodes, on dot blot assays with soluble proteins extracted from a single female. Because of its specificity and sensitivity, the use of the A67 antiserum to improve the diagnosis of the two European quarantine root-knot nematodes is discussed.     

SENSITIVITY AND SPECIFICITY OF RADIOGRAPHIC DETECTION OF CANINE ELBOW INCONGRUENCE IN AN IN VITRO MODEL

Canine elbow incongruence is believed to be the consequence of underdevelopment of the radius. The purpose of this study is to determine the sensitivity and specificity of radiography to detect elbow incongruence in an in vitro model and to assess the optimal elbow angle and radiographic beam position. Five normal cadaveric canine left forelimbs were used. A four-pin, type 1 external fixator with a linear motor side bar was fixed to the cranial part of the radius of each limb and a 1 cm segment of bone was removed from the mid-diaphysis to allow radial shortening. Each elbow was subjected to the same protocol. They were radiographed at two different angles (90 degrees and 135 degrees) of flexion, with 10 different radiographic beam positions (centered on the humeral condyle, 3 cm cranial, 3 cm caudal, 3 cm distal, 3 cm proximal, 3 cm cranio-proximal, 3 cm cranio-distal, 3 cm caudo-proximal, 3 cm caudo-distal and on the shoulder joint) and at four different level of radial shortening (0, 1, 2 and 3 mm). In addition, a radiographic view centered on the elbows flexed at 135 degrees was made after simulating weight bearing. The acquired digital images were independently evaluated by three evaluators unaware of the elbows status. The elbows were judged normal, incongruent or borderline based on specific criteria. The sensitivity for detection of elbow incongruence at and beyond 2 mm was excellent at 90 degrees (median = 100% for all views) and good at 135 degrees (median = 80%) of flexion with no difference between examiners. The sensitivity at 1 mm of incongruence was unchanged at 135 degrees but was reduced at 90 degrees of flexion (median = 60%) with a significant difference between the evaluators. The specificity was significantly different between the evaluators and ranged from 70% to 90% at 90 degrees of flexion and from 50% to 80% at 135 degrees. The lowest specificities at 90 degrees were obtained with the proximal displacements of the X-ray beam. Simulating weight bearing significantly decreased the sensitivity at 1 mm (from 80% to 50%) and 3 mm (from 100% to 80%) of incongruence and slightly increased the specificity (from 55% to 65%). Radiography is a sensitive and specific test to detect moderate-to-severe radio-ulnar incongruence (2 mm and over) if the elbow is flexed at a 90 degrees angle regardless of the radiographic beam position. Finally, canine elbow incongruence appeared reduced after an in vitro weight-bearing simulation.     

MAGNETIC RESONANCE IMAGING APPEARANCE OF SUSPECTED ISCHEMIC MYELOPATHY IN DOGS

Ischemia and infarction of the spinal cord is a known cause of acute spinal injury in dogs. Currently, the diagnosis of spinal cord infarction in small animals is based on history, clinical signs, and the exclusion of other differentials with radiography and myelography. It is a diagnosis only confirmed through necropsy examination of the spinal cord. The aim of this paper is to describe the Magnetic resonance imaging (MRI) findings of the spinal cord of dogs with suspected spinal cord infarcts to utilize this technology for antemortem support of this diagnosis. This retrospective study evaluated the spinal MR examinations of 11 dogs with acute onset of asymmetric nonpainful myelopathies. All patients except one (imaged at 2 months) were imaged within 1 week of clinical signs and managed conservatively with minimal medical and no surgical intervention. They were followed clinically for a minimum of 4 months after discharge. MR findings in all dogs were characterized by focal, intramedullary, hyperintense lesions on T2-weighted images with variable contrast enhancement similar to what is reported in humans. Though it could not be used to diagnose spinal cord infarction definitively, MRI was useful in excluding extramedullary spinal lesions and supporting intramedullary infarction as a cause of the acute neurologic signs. Together with the history and clinical examination findings, MRI is supportive of a diagnosis of spinal cord infarction.     

Plasma Adrenocorticotropin Concentration in Healthy Horses and in Horses With Clinical Signs of Hyperadrenocorticism

Pituitary adenomas are commonly reported in older horses. The typical clinical signs associated with this condition, also known as equine Cushing's disease (ECD), are related to increased adrenocorticotropin (ACTH) production resulting in hyperadrenocorticism. The primary purpose of this study was to determine whether plasma ACTH concentrations differed between cushingoid and healthy horses. The second objective was to determine the effects of blood sample handling techniques on ACTH concentrations. A commercial human ACTH radioimmunoassay (RIA) was used to quantify equine plasma ACTH. Intra-assay and interassay variations, as well as dilutional parallelism were determined during the RIA validation. Plasma ACTH concentrations were evaluated in a group of healthy equids composed of 18 horses and 9 ponies, and in 22 equids with a clinical diagnosis of hyper adrenocorticism (11 horses and 11 ponies). The mean plasma ACTH concentrations in healthy horses and ponies, (18.68 ± 6.79 pg/mL (mean ± SD) and 8.35 ± 2.92 pg/mL, respectively), were significantly different (P = .009). The mean plasma ACTH concentration in horses and ponies with ECD, (199.18 ± 182.82 pglmL and 206.21 ± 319.56 pg/mL, respectively), were significantly higher than the mean ACTH concentration in the control animals (P < .001). Plasma ACTH concentrations appeared to be a sensitive and specific indicator of ECD in horses and ponies. ACTH concentrations measured in plasma samples kept at room temperature (19°C) as long as 3 hours after blood collection were not statistically different from those of samples kept at 1°C. J Vet Intern Med 1996;10:1–6. Copyright 1996 by the American College of Veterinary Internal Medicine .     

Polymorphonuclear proteolytic activity and milk composition change

Relationships between Quarter Milk Cell Count (QMCC) and Tank Milk Cell Count (TMCC) with milk biochemical and technological parameters in milk and dairy products were investigated. All parameters measured were affected by the increase of TMCC and QMCC between 0 and 600 000 cells/mL. The variable effect of lactation stage which is different for different authors, is discussed. The three mechanisms, measured during the inflammation of the udder, implicated in the modification of milk quality are described (a decrease in synthesis, a decrease in the milk barrier permeability and an increase in proteolytic activities). The direct effect of plasmin in caseinolysis is well known; the specific role of the increase of somatic cells (especially PMN) in the modification of milk quality is described. Several specific proteolytic activities of PMN are described and the impact of these activities on caseinolysis is evaluated. Two hypothetical mechanisms of caseinolysis by PMN are suggested and a synthetic scheme of the role of plasmin, bacteria and somatic cells in caseinolysis is discussed.     

Monitoring of atrazine treatment on soil bacterial,fungal and atrazine-degrading communities by quantitative competitive PCR

We report the development of quantitative competitive (QC) PCR assays for quantifying the 16S, 18S ribosomal and atzC genes in nucleic acids directly extracted from soil. QC-PCR assays were standardised, calibrated and evaluated with an experimental study aiming to evaluate the impact of atrazine application on soil microflora. Comparison of QC-PCR 16S and 18S results with those of soil microbial biomass showed that, following atrazine application, the microbial biomass was not affected and that the amount of 16S rDNA gene representing 'bacteria' increased transitorily, while the amount of 18S rDNA gene representing fungi decreased in soil. In addition, comparison of atzC QC-PCR results with those of atrazine mineralisation revealed that, in response to atrazine treatment, the amount of atzC gene increased transitorily in soil pre-treated with atrazine, suggesting that accelerated atrazine biodegradation in soil could be due to a transient increase in the size of the atrazine mineralising community.     

Detection of Salmonella organisms and assessment of a protocol for removal of contamination in horse stalls at a veterinary teaching hospital

OBJECTIVES: To assess methods of detecting environmental contamination with Salmonella organisms and evaluate a cleaning and disinfection protocol for horse stalls in a veterinary teaching hospital. DESIGN: Original study. SAMPLE POPULATION: 37 horses with diarrhea likely to be caused by Salmonella infection and their stall environments. PROCEDURES: Fecal samples were collected from horses daily during hospitalization; samples were obtained from stall sites after cleaning and application of disinfectants. Fecal and environmental samples were cultured for Salmonella spp and tested via polymerase chain reaction (PCR) assay to detect Salmonella DNA. RESULTS: 1 horse died and 2 were discharged prior to sample collection. Fecal samples from 9 of 34 horses yielded growth of Salmonella organisms on bacteriologic culture, and 23 yielded positive results via PCR assay on > or = 1 occasion. Among environmental samples from 21 stalls, salmonellae were detected at > or = 1 stall site on 6 of 78 occasions, and > or = 1 stall site yielded positive results via PCR assay on 69 of 77 occasions. Salmonella DNA was detected more frequently in samples of stall drains, cracks, and corners. Salmonella spp were cultured from samples of 3 stalls after both initial and second cleaning and disinfection cycles, but no organisms were detected in samples obtained after use of a peroxygen disinfectant. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that stalls in which horses with salmonellosis were housed should only be used to accommodate newly hospitalized horses after samples (collected after 2 cycles of cleaning and disinfection) from drains, cracks, and corners yield negative results on bacteriologic culture.     

Thoracoscopic treatment of bullous emphysema in 3 dogs

OBJECTIVE: To report thorascopic partial lobectomy for treatment of bullous emphysema in dogs. STUDY DESIGN: Prospective clinical study. ANIMALS: Three dogs with spontaneous pneumothorax. METHODS: Thoracoscopy without pulmonary exclusion was used to identify bulla. The thorascope was introduced into the thorax lateral to the xyphoid process, and instrument portals were made at different levels along the thoracic wall between the third and tenth intercostal spaces. The thorascope was passed through the mediastinum to view the opposite pleural cavity. After identification of bullae, the affected lung was excised using an endoscopic stapler, and the incision line was checked for air leakage. Thoracic drains were used for air aspiration for 2 days after surgery. RESULTS: Bullae were confirmed histologically as emphysematous lesions. Lung inflation did not interfere with identification of bullae or with surgery. All dogs had full recovery without recurrence for 18 to 29 months after surgery. CONCLUSIONS: Identification and ablation of bulla can be performed thoracoscopically without pulmonary exclusion in dogs. CLINICAL RELEVANCE: Thoracoscopy offers several advantages compared with thoracotomy for treatment and diagnosis of idiopathic pneumothorax, including ease of identification of bullae and reduced postoperative pain and morbidity.     

Gene discovery in Eimeria tenella by immunoscreening cDNA expression libraries of sporozoites and schizonts with chicken intestinal antibodies

Specific antibodies were produced ex vivo from intestinal culture of Eimeria tenella infected chickens. The specificity of these intestinal antibodies was tested against different parasite stages. These antibodies were used to immunoscreen first generation schizont and sporozoite cDNA libraries permitting the identification of new E. tenella antigens. We obtained a total of 119 cDNA clones which were subjected to sequence analysis. The sequences coding for the proteins inducing local immune responses were compared with nucleotide or protein databases and with expressed sequence tags (ESTs) databases. We identified new Eimeria genes coding for heat shock proteins, a ribosomal protein, a pyruvate kinase and a pyridoxine kinase. Specific features of other sequences are discussed.     

Quantitative PCR monitoring of the effect of azoxystrobin treatments on Mycosphaerella graminicola epidemics in the field

Quantitative PCR and visual monitoring of Mycosphaerella graminicola epidemics were performed to investigate the effect of curative and preventative applications of azoxystrobin in wheat field crops. A non-systemic protectant and a systemic curative fungicide, chlorothalonil and epoxiconazole, respectively, were used as references. PCR diagnosis detected leaf infection by M graminicola 3 weeks before symptom appearance, thereby allowing a clear distinction between curative and preventative treatments. When applied 1 week after the beginning of infection, azoxystrobin curative activity was intermediate between chlorothalonil (low effect) and epoxiconazole. When applied preventatively, none of the fungicides completely prevented leaf infection. There was some indication that azoxystrobin preventative treatments may delay fungal DNA increase more than epoxiconazole at the beginning of leaf infection. Both curative and preventative treatments increased the time lapse between the earliest PCR detection and the measurement of a 10% necrotic leaf area. Azoxystrobin only slightly decreased the speed of necrotic area increase compared with epoxiconazole. Hence, azoxystrobin activity toward M graminicola mainly resides in lengthening the time lapse between the earliest PCR detection and the measurement of a 10% necrotic leaf area. Information generated in this way is useful for optimal positioning of azoxystrobin treatments on M graminicola.