Expression of erythropoietin in cats treated with a recombinant adeno-associated viral vector

OBJECTIVE: To characterize the biological effects of IM administration of a recombinant adeno-associated virus serotype 2 (rAAV2) vector containing feline erythropoietin (fEPO) cDNA and determine whether readministration of the vector or removal of muscle tissue at the injection sites alters those effects. ANIMALS: 10 healthy 7-week-old specific pathogen-free cats. PROCEDURE: Cats received 1 X 10(7) infective units (iU; n = 3), 1 X 10(8) iU (3), or 1 X 10(9) iU (2) of rAAV2-fEPO vector IM (day 0). Two control cats received an rAAV2 vector containing the LacZ gene (1 X 10(9) iU, IM). In all cats, hematologic variables and serum fEPO concentration were measured at intervals; anti-rAAV2 antibody titer was measured on day 227. In cats that did not respond to treatment, the rAAV2-fEPO vector was readministered. Injection sites were subsequently surgically removed. RESULTS: Compared with control cats, cats treated with 1 X 10(9) iU of rAAV2-fEPO vector had increased Hct and serum fEPO concentrations. One of these cats developed pure RBC aplasia; its Hct normalized following injection site excision. Cats receiving lower doses of vector had no response; on retreatment, 1 of those cats developed sustained erythrocytosis that persisted despite injection site removal and the others did not respond or responded transiently. Antibodies against rAAV2 were detected in all vector-treated cats. CONCLUSIONS AND CLINICAL RELEVANCE: Gene therapy may be an effective treatment for cats with hypoproliferative anemia. However, rAAV2-fEPO vector administration may result in pure RBC aplasia or pathologic erythrocytosis, and injection site removal does not consistently abolish the biological response.     

Encrusted cystitis secondary to Corynebacterium matruchotii infection in a horse

A 17-year-old gelding was evaluated because of dysuria, inappetence, and weight loss. Cystoscopy revealed severe mucosal ecchymoses with luminal hemorrhage and accumulations of crystalloid sludge. Analysis of a urine sample revealed isosthenuria, an alkaline pH, pyuria, hematuria, bacteriuria, and numerous calcium carbonate crystals. Histologic examination of bladder mucosa biopsy specimens revealed severe neutrophilic infiltration with mineralization. A diagnosis of encrusted cystitis exacerbated by sabulous urolithiasis was made. A Corynebacterium sp susceptible to penicillin, sulfonamide, and enrofloxacin was cultured from the urine and the bladder mucosa biopsy specimens. The horse was treated with penicillin G potassium, IV, for 5 days, followed by trimethoprim-sulfamethoxazole for 4 weeks. Bladder lavage was performed daily for the first 3 days with a balanced electrolyte solution and dimethyl sulfoxide in an attempt to aid expulsion of necrotic debris and crystalline sludge from the bladder. Molecular phylogenetic analysis based on the 16S rDNA gene sequence was used to identify the isolate and determine its phylogenetic position. Results indicated that the isolate was closely related to Corynebacterium matruchotii. To our knowledge, encrusted cystitis secondary to C matruchotii has not been previously identified in a horse.     

Correlation between blood and milk serum leptin in goats and growth of their offspring

Boer and Boer crossbred meat-type does were used in two experiments to determine whether goat milk serum contains leptin and to investigate possible correlations of milk and serum leptin in does and subsequent growth of their offspring. Blood and milk samples were collected within 2 h of kidding (d 0) from 20 (Exp. 1; spring) or 22 does (Exp. 2; the following fall). Blood milk samples were then collected again on d 0.5, 1, 3, 5, 7, 14, 21, 28, 35, 42, 49, and 56 (Exp. 1) or d 0.5, 1, 2, 3, 4, 5, 6, 7, 14, and 21 (Exp. 2). Body weights of kids were recorded on d 0, and BW of kids and does were recorded weekly beginning on d 7 (kids) or 21 (does), with BCS also recorded for does beginning on d 28 for Exp. 1 and on d 0.5, 1, 2, 3, 4, 5, 6, 7, 14, and 21 for Exp. 2. Leptin was detected in colostral milk and was influenced by days postpartum, decreasing (P < 0.001) over time with an average of 4.4 +/- 0.3 ng/mL (Exp. 1) and 18.1 +/- 1.0 ng/mL (Exp. 2) on d 0 compared with 1.0 +/- 0.3 ng/mL on d 56 (Exp. 1) and 2.9 +/- 0.2 ng/mL on d 21 (Exp. 2). Day postpartum and milk serum leptin were negatively correlated (P < 0.001) for Exp. 1 (r = -0.27) and Exp. 2 (r = -0.46). For Exp. 1 only, blood serum leptin tended (P = 0.09) to be influenced by day, with a weak positive correlation (r = 0.15; P < 0.02). Weak positive correlations (P < 0.01) were found between blood serum leptin and doe BCS (r = 0.42 in Exp. 1, and r = 0.13 in Exp. 2) and doe BW (r = 0.44 in Exp. 1, and r = 0.26 in Exp. 2), with the absence of a stronger relationship likely due in part to the short time period measured and the lack of significant changes in BCS and BW during that time. In conclusion, leptin was present in milk and blood serum of does, and blood serum leptin was weakly correlated with doe BW and BCS, but it was not related to kid BW. Therefore, further studies are needed to clarify the relationships involving milk and serum leptin in goats.     

Proximal tibial intraarticular ostectomy for treatment of canine cranial cruciate ligament injury

OBJECTIVE: To report a technique for surgical alteration of the slope of the tibial plateau by a proximal tibial intraarticular ostectomy (PTIO) after injury to the canine cranial cruciate ligament (CCL) and to determine the outcome. STUDY DESIGN: Prospective clinical study. ANIMALS: Dogs (n=52) with CCL injury in 60 stifle joints. METHODS: CCL injury was treated by lateral stifle arthrotomy, removal of CCL remnants, and appropriate meniscal surgery. PTIO was performed to remove a wedge of bone from the proximal aspect of the tibia. The ostectomy site was reduced and stabilized using a bone plate and screws applied to the medial surface of the tibia as well as a craniocaudal positional screw. Dogs were evaluated at 6 weeks, 6, and 12 months by complication assessment, lameness scores, stifle range of motion (ROM), thigh circumference, radiographic assessment, degenerative joint disease (DJD) scores, and surgeon and owner evaluation of function. RESULTS: Lameness scores improved by 6 and 12 months in all but 1 dog. Thigh circumference and DJD were increased at 6 and 12 months. Complications occurred in 20% of dogs with all but 1 occurring perioperatively or within 6 weeks; most common were injury to the long digital extensor tendon (4 dogs) and plate failure (3); 2 other dogs required surgery to treat complications. Most owners (98%) reported that lameness had improved by 12 months; 90% were extremely or very satisfied with the procedure and 90% would have the same procedure performed on another dog. CONCLUSION: PTIO to level the tibial plateau provided a satisfactory clinical outcome in dogs >20 kg with CCL injury and the complication rate was similar to tibial plateau levelling osteotomy (TPLO). Stifle osteoarthritis continued to progress radiographically. CLINICAL RELEVANCE: PTIO represents an alternative to TPLO that does not require specialized surgical equipment.     

Immunohistochemical identification of Campylobacter fetus in natural cases of bovine and ovine abortions

An immunohistochemistry (IHC) procedure for the detection of Campylobacter fetus antigens using an avidin-biotin complex technique was performed on formalin fixed bovine and ovine fetal tissues from 26 natural cases of Campylobacter spp. abortion (four ovine and 22 bovine). The species of Campylobacter isolated included C. fetus ssp. venerealis from 13 bovine fetuses, C. fetus ssp. fetus from two ovine and one bovine fetus, Campylobacter jejuni from seven bovine fetuses, Campylobacter lari from two ovine fetuses and an unspeciated Campylobacter species in one bovine fetus. Histologic lesions identified in the aborted fetuses included placentitis, serositis, pneumonia, gastroenteritis, hepatitis and encephalitis. Campylobacter fetus antigens were identified by IHC in 13 of 13 bovine fetuses from which C. fetus ssp. venerealis was isolated and in two of two ovine fetuses from which C. fetus ssp. fetus was isolated. The IHC stains were negative in tissues from seven bovine fetuses from which C. jejuni was isolated, one bovine fetus infected with C. fetus ssp. fetus, one bovine fetus infected with the unspeciated Campylobacter and two ovine fetuses infected with C. lari. In positive cases, the IHC stain most frequently identified bacteria in the lung and gastrointestinal tract. The C. fetus IHC procedure performed on formalin fixed tissues is a practical tool for the diagnosis of natural cases of ovine and bovine abortion caused by C. fetus.     

Risk factors for Clostridium piliforme infection in foals

OBJECTIVE: To determine risk factors for Clostridium piliforme infection in neonatal foals on a Thoroughbred breeding farm in California. DESIGN: Case-control and retrospective cohort studies. ANIMALS: 322 neonatal Thoroughbred foals either born on the study farm or born elsewhere but traveled to the farm with their dam during the 1998, 1999, and 2000 breeding seasons. PROCEDURE: Mare and foal records from 1998, 1999, and 2000 were examined, using case-control design methods to determine variables associated with increased risk of C. piliforme infection in foals. Important risk factors identified in the case-control study were then reevaluated by use of a retrospective cohort design, using data from all neonatal foals present on the farm during the 3-year study period. RESULTS: Foals born between March 13 and April 13 were 7.2 times as likely to develop C. piliforme infection as were those born at any other time of the foaling season. Foals of nonresident (visiting) mares were 3.4 times as likely to develop disease as were foals born to mares that were permanent residents of the study farm. Foals of mares < 6 years of age were 2.9 times as likely to develop disease as were foals born to older mares. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this research can be used to better understand the epidemiologic factors of C. piliforme infection in horses. High-risk foals can be closely monitored to aid in early diagnosis and treatment, resulting in the best possible clinical outcome for affected individuals.     

Seeking a niche: putative contributions of the hfq and bacA gene products to the successful adaptation of the brucellae to their intracellular home

Long-term residence of the brucellae in the phagosomal compartment of host macrophages is essential to their ability to produce disease in both natural and experimental hosts. Correspondingly, the Brucella spp. appear to be well adapted to resist the multiple environmental stresses they encounter in their intracellular home. This brief review will focus on the contributions of the hfq and bacA gene products to this adaptation. Studies with Brucella hfq mutants suggest that stationary phase physiology is critical for successful long-term residence in host macrophages. Analysis of Brucella bacA mutants, on the other hand, reveal very striking parallels between the strategies employed by the rhizobia to establish and maintain protracted intracellular residence in their plant host and those used by the brucellae during their long-term survival in the phagosomal compartment of host macrophages.     

Functional connectivity defined through cost-distance and genetic analyses: a case study for the rock-dwelling mountain vizcacha (<Emphasis Type="Italic">Lagidium viscacia</Emphasis>) in Patagonia,Argentina

Landscape connectivity can have profound consequences for distribution and persistence of populations and metapopulations. Evaluating functional connectivity of a landscape for a species requires a measure of dispersal rates through landscape elements at a spatial scale sufficient to encompass movement capabilities of individuals over the entire landscape. We evaluated functional connectivity for a rock-dwelling mammal, the mountain vizcacha (Lagidium viscacia), in northern Patagonia. Because of the strict association of mountain vizcachas with rocks, we hypothesized that connectivity for this species would be influenced by geology. We used molecular genetic estimates of gene flow to test spatially explicit models of connectivity created with GIS cost-distance analysis of landscape resistance to movement. We analyzed the spatial arrangement of cliffs with join counts and local k-function analyses. We did not capture and genotype individuals, but sampled at the population level through non-invasive collection of feces of mountain vizcachas. The model of landscape connectivity for mountain vizcachas based on geology was corroborated by the pattern of genetic structure, supporting the hypothesis that functional connectivity for mountain vizcachas is influenced by geology, particularly by the distribution of appropriate volcanic rocks. Analysis of spatial arrangement of cliffs indicated that occupied cliffs are clustered and confirmed that rivers act as barriers to dispersal for mountain vizcachas. Our methods could be used, within certain constraints, to study functional landscape connectivity in other organisms, and may be particularly useful for cryptic or endangered species, or those that are difficult or expensive to capture.     

Histochemical and immunohistochemical evidence of a bacterium associated with lesions of epizootic bovine abortion.

Epizootic bovine abortion (EBA), a tick-transmitted disease of pregnant cattle grazing foothill pastures, is a major cause of reproductive failure in California and adjacent states. Affected fetuses develop a chronic disease, resulting in late-term abortion or premature calving. Despite investigations spanning 50 years, to the authors' knowledge, the etiologic agent of EBA has not yet been isolated from affected fetuses or the tick vector. The diagnosis of EBA is based on gross and microscopic lesions. Recently, documentation that the etiologic agent is susceptible to antibiotics and identification of a unique 16S deltaproteobacterial rDNA gene sequence in 90% of thymus tissues from aborted fetuses have supported the role of a bacterial infection as the cause of EBA. To determine whether bacteria could be detected in the tissues, histochemical staining and immunohistochemical procedures were used on formalin-fixed, paraffin-embedded tissues. Use of a modified Steiner silver stain revealed small numbers of intracytoplasmic bacterial rods in 37 of 42 thymic samples from EBA-affected fetuses. Improved detection was achieved by use of immunohistochemical staining with serum from EBA-affected fetuses that resulted in detection of numerous bacterial rods in the cytoplasm of histiocytic cells in the thymus from all 42 EBA-affected fetuses. Immunohistochemical examination of additional tissues from 21 field and experimental EBA cases revealed positively stained intracytoplasmic bacterial rods in many organs with inflammatory lesions. Use of the modified Steiner stain and immunohistochemical staining of tissues from negative-control fetuses failed to reveal organisms. To the authors' knowledge, this is the first report to document morphologic evidence of a bacterium associated with the lesions of EBA.