Immunogenicity of bovine parainfluenza type 3 virus proteins encapsulated in nanoparticle vaccines,following intranasal administration to mice

Stimulation of long lasting, protective immunity to respiratory viruses is often difficult to achieve with conventional respiratory vaccines. Polymeric nanoparticles, incorporating viral proteins have been shown to offer sustained release of antigen, with consequent prolongued stimulation of the respiratory immune system. In this paper the efficacy of two nanoparticle vaccines (poly-lactide-co-glycolide, PLGA; polymethylmethacrylate, PMMA), incorporating proteins of bovine parainfluenza type 3 virus (BPI-3) was investigated. As a preliminary to experiments in calves, it was considered essential to demonstrate immunogenicity of the experimental vaccine in mice. Mice immunised with PLGA nanoparticles, containing BPI-3 proteins, developed higher levels of virus-specific antibody than mice immunised with the PMMA vaccine or with soluble viral proteins alone. Immunoblotting using serum from the vaccinated mice, demonstrated strong reactions against the major BPI-3 proteins.     

In vitro effects of osteotomy angle and osteotomy reduction on tibial angulation and rotation during the tibial plateau-leveling osteotomy procedure

OBJECTIVE: To determine the effect of osteotomy angle, reduction technique, and tibial plateau rotation angle on angular and rotational limb deformities. STUDY DESIGN: Geometric comparison using bone models. METHODS: Rotational osteotomies were made in the proximal metaphysis of artificial tibias at 0 degrees, 10 degrees, 20 degrees, -10 degrees, and -20 degrees from perpendicular with respect to either the proximodistal and craniocaudal tibial axes. Negative-numbered angles represented osteotomies made from distal to proximal or caudal to cranial. Changes in tibial angulation and torsion were measured using a 3-dimensional digitizing instrument at tibial plateau rotation angles from 0 degrees to 30 degrees at 5 degrees increments. Two osteotomy reduction techniques were used: complete osteotomy reduction and alignment of the medial cortex. The mean of 5 measurements of torsional and angular tibial deformity for each of the 9 osteotomy orientations in each reduction technique group was obtained. RESULTS: All had increasing angular and rotational deformity as tibial plateau rotation angle increased. In the medially aligned cortex group, all tibias had valgus deformity, and 8 of 9 tibias were internally rotated. In the reduced osteotomy group, minimal angular deformity was seen in tibias with osteotomy variation along the proximodistal axis; however, tibias with osteotomy variation along the craniocaudal axis had angular deformity ranging from 6.0 degrees of varus deformity to 14.3 degrees of valgus deformity. Rotational deformity was affected similarly by osteotomy variation along either axis. Reduction technique had greater affect on angular and rotational deformity than osteotomy angle variation. CLINICAL RELEVANCE: These results suggest that osteotomy reduction may play a greater role in angular and rotational deformity than osteotomy angle, although extreme osteotomy angles should be avoided. To decrease the severity of deformity, we recommend that the osteotomy be made perpendicular to the craniocaudal and proximodistal axes and be completely reduced with less regard for alignment of the medial cortex.     

Circular external skeletal fixation stabilization of antebrachial and crural fractures in 25 dogs

Fracture stabilization using circular external skeletal fixation was evaluated in 14 dogs with antebrachial fractures and 11 dogs with crural fractures. Most dogs were consistently weight bearing on the stabilized limb by 3 days following surgery. Although all dogs developed minor wire/pin tract inflammation and eight dogs developed major wire/pin tract inflammation, postoperative lameness was not consistently associated with wire/pin tract complications. Fractures in 23 dogs achieved radiographic union (mean +/- standard deviation [SD], 61 +/- 21 days; median, 57 days) without additional surgery; two dogs required restabilization of their fractures with linear fixators. Twenty of the 21 owners that could be contacted felt their dog had no (n=15) or only a mild (n=5) intermittent lameness at the time of final, long-term (mean +/- SD, 37 +/- 17 months; median, 42 months) assessment.     

Use of pneumocystoplasty for overinflation of the swim bladder in a goldfish

A Ryukin goldfish was evaluated because of a 6-month history of progressive abdominal distention and positive buoyancy. Overinflation of the swim bladder was diagnosed, and the fish was anesthetized with tricaine methanesulfonate. Archimedes' principle was used to determine the volume of swim bladder that was removed surgically. The caudal swim bladder was exteriorized through an abdominal incision and 2 surgical clips were placed across it to limit its size. After surgery, the fish remained in a state of negative buoyancy in sternal and lateral recumbency on the bottom of the tank. Sutures were removed 15 days after surgery, but the fish died 24 days after surgery. A full necropsy could not be performed because of autolysis of the tissues, but the surgical clips and the swim bladder appeared unremarkable. Pneumocystoplasty may be a viable treatment for this condition.     

Clinical and hematologic effects of experimental infection of dogs with recently identified Babesia gibsoni-like isolates from Oklahoma

OBJECTIVE: To characterize clinical and hematologic responses in dogs following experimental inoculation with Babesia gibsoni-like isolates from infected dogs in Oklahoma. DESIGN: Prospective study. ANIMALS: 6 mixed-breed dogs. PROCEDURE: 2 dogs were inoculated with organisms from a naturally infected dog, and 3 were inoculated with organisms from a second naturally infected dog (1 of these 3 dogs was splenectomized 1 week prior to inoculation). One dog was not inoculated. Complete blood counts were performed weekly. RESULTS: In the 5 dogs inoculated with organisms, parasites were initially detected 1 to 5 weeks after inoculation, and severity of parasitemia peaked with 1.9 to 6.0% of RBC infected by 4 to 6 weeks after inoculation. Parasitemia was easily detectable (> 0.1% of RBC infected) for 3 to 4 weeks. Clinical abnormalities included lethargy, fever, and pale mucous membranes but were mild to nearly inapparent in 2 dogs. All dogs developed regenerative anemia and marked thrombocytopenia. Thrombocytopenia developed before and lasted longer than the parasitemia. Profound but transient neutropenia was detected in some dogs. The splenectomized dog developed more severe parasitemia and anemia and more pronounced clinical abnormalities. Three dogs with intact spleens recovered without treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that 2 or more genotypically distinct, but morphologically identical, small Babesia parasites can infect dogs in the United States. Compared with infection with small Babesia parasites from California, infection with these isolates resulted in less severe parasitemia and clinical abnormalities. Parasitemia was transient, indicating that identification of organisms in blood smears may be difficult in some dogs.     

Evaluation of intravenous administration of meloxicam for perioperative pain management following stifle joint surgery in dogs

OBJECTIVE: To compare preoperative administration of meloxicam and butorphanol to perioperative administration of butorphanol alone for control of postoperative signs of pain in dogs. ANIMALS: 40 client-owned dogs scheduled for surgical repair of a cranial cruciate ligament rupture. PROCEDURE: Group-1 dogs received butorphanol (0.2 mg/kg, IV) and meloxicam (0.2 mg/kg, IV) just prior to surgery. Group-2 dogs received butorphanol just prior to surgery (0.2 mg/kg, IV) and at incision closure (0.1 mg/kg, IV). Pain assessment began 1 to 2 hours before surgery and from extubation until 24 hours after surgery by obtaining the following measurements: the visual analog scale (VAS) score, cumulative pain score (CPS), adjusted cumulative pain score, modified cumulative pain score, and the adjusted modified cumulative pain score (AMCPS). Serum cortisol concentration was measured between 12 to 24 and between 1 to 2 hours prior to surgery, and at 30 minutes, and 1, 2, 4, 8, 18, and 24 hours after extubation. RESULTS: No significant differences between treatment groups were observed in CPS or VAS score. At 8, 9, 10, and 11 hours after extubation, meloxicam-butorphanol-treated dogs had a significantly lower AMCPS, compared with butorphanol-alone-treated dogs. Total serum cortisol concentration (area under the curve) during the measurement period was significantly lower in meloxicam-butorphanol-treated dogs, compared with butorphanol-alone treated dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Preoperative single dose administration of meloxicam-butorphanol is equivalent to or slightly better than the administration of 2 perioperative doses of butorphanol for the control of postoperative signs of pain in dogs.     

Identification of major allergens of Malassezia pachydermatis in dogs with atopic dermatitis and Malassezia overgrowth

Abstract We have previously shown that both atopic and normal dogs generate an IgG response to antigens of Malassezia pachydermatis . The aim of this study was to compare IgE responses to separated proteins of M. pachydermatis in 28 atopic dogs with Malassezia dermatitis and 22 clinically normal dogs using Western immunoblotting. Six different detection systems were evaluated in order to assess sensitivity and eliminate nonspecific binding and cross-reactivity. The protocol yielding the best results utilized a monoclonal mouse antidog IgE, an alkaline phosphatase conjugated goat antimouse IgG which had been passed through a canine IgG column 3 times, a chemiluminescent substrate and a digital imaging system. Proteins of 45, 52, 56 and 63 kDa were recognized by more than 50% of the atopic dog sera and thus represented major allergens. Only a minority of normal dogs showed faint IgE binding to these proteins. The results indicate that the majority of atopic dogs with Malassezia dermatitis have a greater IgE response than normal dogs, suggesting an IgE-mediated immune response may be clinically important in the pathogenesis of the disease.     

Temporal and spatial dynamics of insecticide resistance in Myzus persicae (Hemiptera: Aphididae)

Advances in understanding insecticide resistance in the peach-potato aphid, Myzus persicae (Sulzer), at the genotypic, biochemical and molecular levels have led to rapid and precise methods for the detection of several resistance mechanisms (elevated carboxylesterase, modified acetylcholinesterase or MACE, and knockdown resistance or kdr) in individual insects, and for monitoring their frequencies over space and time. This paper summarises the results of two long-term surveys of resistance dynamics in M persicae in England, based on samples collected directly from field and glasshouse crops or from four 12.2-m suction traps. The study showed marked fluctuations in resistance frequencies that probably reflect the counteracting forces of selection imposed by insecticides for aphids possessing more copies of esterase resistance genes, and selection against these forms when insecticide use is relaxed. There is growing evidence that several different resistance mechanisms in M persicae have associated fitness costs. In the case of esterase and MACE, these costs are apparently strong enough to effect a decline in resistance frequency over winter, and a more prolonged decline over successive cropping seasons when aphid numbers are insufficient to trigger intensive chemical applications. Changes in the overall frequency of resistance genotypes may also be influenced by the predominance of year-round parthenogenesis in M persicae in the UK, leading to non-random associations between mechanisms and selection operating on clonal lineages rather than individual genotypes.     

Phenotypic characterisation of Phaeoacremonium and Phaeomoniella strains isolated from grapevines: enzyme production and virulence of extra-cellular filtrate on grapevine calluses

Extra-cellular enzyme production of different Phaeoacremonium spp. and Phaemoniella chlamydospora isolates were used to assay the possibility of inter-specific characterisation. Isolates of Phaeoacremonium aleophilum, Phaeoacremonium angustius, P. viticola and Ph. chlamydospora were grown on solid media and the activities of extra-cellular amylases, lipases, proteases, cellulases, xylanases, laccase, polygalacturonase, pectate lyase, lignin peroxidase, manganese peroxidase, urease and chitinase were assayed. Phaeoacremonium species showed activities of a larger number of enzymes and also enzyme activity was frequently higher suggesting that Phaeoacremonium can be more virulent. To assay if the produced extra-cellular enzymes could reflect the virulence capacity of the two genera, calluses of Vitis vinifera L. (cvs. Baga and Maria Gomes) and of a rootstock (R3309) were inoculated with filtrated culture liquid medium of three isolates of Ph. chlamydospora and one of P. angustius. Filtrates from all strains decreased callus growth and membrane integrity, while soluble protein content of calluses decreased with the strains CAP 054 and 1AS. P. angustius (CAP 054) induced the more severe symptoms in all genotypes. Water content decreased together with an increase of osmolality in both cultivars but not in rootstock suggesting that osmorregulatory capacity is more affected in cultivars. Data show that: (1) Phaeoacremonium and Phaeomoniella genera have different patterns of extra- cellular enzymatic production; (2) these fungi produce extra-cellular compound(s) that induce(s) senescence symptoms in plant cells inhibiting callus proliferation; (3) among the strains tested in plant calluses the most virulent isolate (CAP 054) also produced higher amounts of some extra-cellular enzymes; (5) rootstock calluses were less sensitive to inoculation than grapevine calluses.