Prevalence of Bartonella sp. in United States military working dogs with infectious endocarditis: a retrospective case-control study.

OBJECTIVES: Bartonella infection has been associated with endocarditis in humans, dogs, cats and cattle. In order to evaluate the importance of this pathogen as a possible source of endocarditis in United States military working dogs (MWDs), we performed a retrospective case-control study on 26 dogs with histological diagnosis of culture negative endocarditis (n = 18), endomyocarditis (n = 5) or endocardiosis (n = 3) and 28 control dogs without any histological cardiac lesions. METHODS: DNA was extracted from paraffin embedded cardiac valves and tissues from case and control dogs and submitted to PCR testing with primers targeting the Bartonella gltA gene. PCR-RFLP using four restriction endonucleases and partial sequencing was then performed to determine the Bartonella species involved. RESULTS: Nineteen (73%) cases were PCR positive for Bartonella, including B. henselae (8 dogs), B. vinsonii subsp. berkhoffii (6 dogs), B. washoensis (2 dogs) and B. elizabethae (1 dog). Only one control dog was weakly PCR positive for Bartonella. Based on the type of histological diagnosis, 13 (72.2%) dogs with endocarditis, 3 (60%) dogs with endomyocarditis and all 3 dogs with endocardiosis were Bartonella PCR positive. CONCLUSIONS: Bartonella sp. Infections were correlated with cardiopathies in US military working dogs. Systemic use of insecticides against ectoparasites and regular testing of MWDs for Bartonella infection seem highly appropriate to prevent such life-threatening exposures.

Seroepidemiology of Bartonella vinsonii subsp berkhoffii exposure among healthy dogs.

OBJECTIVE: To determine seroprevalence of antibodies to Bartonella vinsonii subsp berkhoffii and risk factors for seropositivity among working dogs owned by the US government. DESIGN: Cross-sectional study. ANIMALS: 1,872 dogs. PROCEDURE: An ELISA was used to detect antibodies to B vinsonii subsp berkhoffii. RESULTS: Antibodies to B vinsonii subsp berkhoffii were detected in 162 dogs (8.7%; 95% confidence interval, 7.4 to 10.0%). Dogs living in the southeast, plains states, southwest, and south-central were significantly more likely to be seropositive than were dogs living in other regions of the United States. German Shepherd-type dogs were significantly less likely to be seropositive than were dogs of other breeds, and dogs entering training programs or that had been rejected from a training program were significantly more likely to be seropositive than were dogs used for narcotics detection and dogs trained to patrol or detect explosives. Dogs used by the border patrol or Federal Aviation Administration were more likely to be seropositive than were dogs used by the Department of Defense or customs service. Odds that dogs would be seropositive were significantly higher for dogs stationed in the southern United States, the northeastern United States, or a foreign country, compared with dogs stationed in all other regions of the United States. CONCLUSIONS AND CLINICAL RELEVANCE: Overall, 8.7% of this diverse group of healthy dogs was found to be seropositive for antibodies to B vinsonii subsp berkhoffii, and seropositivity rates were associated with location, suggesting either that there are multiple vectors for the organism or that the major vector for the organism depends on geographic and environmental factors.

Evaluation of butylated hydroxytoluene as a cryopreservative added to whole or skim milk diluent for bull semen.

Effect of five concentrations of butylated hydroxytoluene on the percent motility and percent intact acrosomes was evaluated for bull sperm frozen and thawed in whole and skim milk diluents. Twelve ejaculates were frozen in .5-ml straws in moving nitrogen vapor after each ejaculate was split among the five butylated hydroxytoluene treatments within the two diluents. Prefreeze and postthaw percent motility and postthaw percent intact acrosomes were evaluated at 0 and 4 h. Before freezing, sperm motilities were similar in whole or skim milk diluents with and without butylated hydroxytoluene. Upon thawing, sperm motility was 10 percentage points higher in whole milk diluent containing .5 and .75 mM butylated hydroxytoluene than in samples without it. After 4 h, percentage motile sperm was similar among all whole milk treatments. Sperm motility was similar among all skim milk treatments at both 0 and 4 h. Percent intact acrosomes were similar among all treatments for each diluent. Oxidation in whole milk diluent stored at 4 degrees C for 1 wk was reduced in the presence of 1.0, 2.0, and 4.0 mM butylated hydroxytoluene.

Cryopreservation of murine embryos with trehalose and glycerol.

Several concentrations of trehalose (0.0, 0.04, 0.1, 0.25 M) in combination with three concentrations of glycerol (1.0, 1.5, 2.0 M) were evaluated for the cryopreservation of murine embryos. Embryos were transferred through increasing concentrations of glycerol in Dulbecco's phosphate-buffered saline with 10% fetal calf serum (PBS + FCS) to reach the final glycerol concentrations. They were then randomly assigned to one of the concentrations of trehalose. A total of 506 morulae were packaged individually in 0.25-ml plastic straws and cooled from ambient temperature at 1.0 degrees C/min in a programmable methanol freezer. Embryos were seeded at -7 degrees C and then cooled to -25 degrees C at 0.3 degrees C/min before being plunged into liquid nitrogen. After thawing and a one-step dilution of glycerol, embryos were cultured for 48 hr and viability was determined by blastocoel formation. Highest viability (70.0%) after 48 hr in culture was obtained for embryos frozen in 1.5 M glycerol plus 0.10 M trehalose as compared to 31% viability for embryos frozen with glycerol alone. These observations suggest that trehalose can be used in combination with glycerol as a cryoprotectant and that a high rate of viability can be achieved after a one-step dilution of the cryoprotectants.