Quality control ranges for testing broth microdilution susceptibility of Flavobacterium columnare and F. psychrophilum to nine antimicrobials.

A multi-laboratory broth microdilution method trial was performed to standardize the specialized test conditions required for the fish pathogens Flavobacterium columnare and F. psychrophilum. Nine laboratories tested the quality control (QC) strains Escherichia coli ATCC 25922 and Aeromonas salmonicida subsp. salmonicida ATCC 33658 against 10 antimicrobials (ampicillin, enrofloxacin, erythromycin, florfenicol, flumequine, gentamicin, ormetoprim/sulfadimethoxine, oxolinic acid, oxytetracycline, and trimethoprim/sulfamethoxazole) in diluted (4 g l-1) cation-adjusted Mueller-Hinton broth incubated at 28 and 18°C for 44-48 and 92-96 h, respectively. QC ranges were set for 9 of the 10 antimicrobials. Most of the minimal inhibitory concentration (MIC) distributions (16 of 18, 9 drugs at both temperatures) for A. salmonicida ATCC 33658 were centered on a single median MIC ± 1 two-fold drug dilution resulting in a QC range that spanned 3 dilutions. More of the E. coli ATCC 25922 MIC distributions (7 of 16) were centered between 2 MIC dilutions requiring a QC range that spanned 4 dilutions. A QC range could not be determined for E. coli ATCC 25922 against 2 antimicrobials at the low temperature. These data and their associated QC ranges have been approved by the Clinical and Laboratory Standards Institute (CLSI), and will be included in the next edition of the CLSI M49-A Guideline. This method represents the first standardized reference method for testing fish pathogenic Flavobacterium spp.

Comparison of transvaginal natural orifice transluminal endoscopic surgery (NOTES®) and laparoscopy for elective bilateral ovariectomy in standing mares.

OBJECTIVE: To compare surgical trauma, perioperative pain, surgical time, and complication rate in mares undergoing standing bilateral ovariectomy by natural orifice transluminal endoscopic surgery (NOTES) or laparoscopy. STUDY DESIGN: Experimental study. ANIMALS: Healthy mares (n = 12). METHODS: Ovariectomy was performed with a vessel-sealing device by transvaginal NOTES (n = 6) using a flexible endoscope and specialized instruments or by bilateral flank laparoscopy (n = 6). Preoperative and postoperative complete blood count (CBC), plasma fibrinogen concentration, serum amyloid A concentration, peritoneal nucleated cell count, and total protein concentration were compared using repeated measures ANOVA. Surgical times were compared using an unpaired t-test. Mares were monitored for 2 weeks postoperatively and necropsy was performed at 3 weeks (n = 6) or 3 months (n = 6) to assess short- and medium-term effects of each procedure. Complication rate, necropsy, and bacteriology findings were compared using a Fishers exact test. Significance was set at P < .05. RESULTS: All surgical procedures were performed successfully and all but one laparoscopy mare were bright, alert, and maintained a good appetite until euthanasia. Mean ± SD surgical time was similar between groups (NOTES: 100 ± 40 minutes; laparoscopy: 107 ± 47 minutes). There were no significant temporal differences in vital parameters, CBC, plasma fibrinogen concentration, serum amyloid A concentration, peritoneal nucleated cell count, and total protein concentration between groups. CONCLUSIONS: Bilateral ovariectomy by transvaginal NOTES or laparoscopy resulted in minimal inflammation and surgical trauma.

Leaching of Mycobacterium avium Subsp paratuberculosis in Soil under In Vitro Conditions.

Mycobacterium avium subsp paratuberculosis (Map), the causative agent of Johne's disease, has a robust ability to survive in the environment. However, the ability of Map to migrate through soil to drainage tiles or ground water, leave the farm, and leak into local watersheds is inadequately documented. In order to assess the ability of Map to leach through soil, two laboratory experiments were conducted. In the first study, 8 columns (30 cm long each) of a sandy loam soil were treated with pure cultures of Map. Two soil moisture levels and two Map concentrations were used. The columns were leached with 500 mL of water once a week for three weeks, the leachate was collected, and detection analysis was conducted. In the second experiment, manure from Map negative cows (control) and Map high shedder cows (treatment) were deposited on 8 similar columns and the columns were leached with 500 mL of water once a week for four weeks. Map detection and numeration in leachate samples were done with RT-PCR and culture techniques, respectively. Using RT-PCR, Map could be detected in the leachates in both experiments for several weeks but could only be recovered using culture techniques in experiment one. Combined, these experiments indicate the potential for Map to move through soil as a result of rainfall or irrigation following application.

Serologic survey for selected infectious diseases in raccoons (Procyon lotor) in Indiana, USA.

The objective of this study was to characterize the antibody prevalence of important livestock and domestic animal pathogens in raccoons (Procyon lotor) trapped and sampled in 39 forest patches in north-central Indiana, USA, between 2004 and 2005. A total of 459 serum samples were tested for antibodies to Leptospira serovars, 512 for Canine distemper virus (CDV) antibodies, and 340 for antibodies to Porcine reproductive and respiratory syndrome virus (PRRSV). In total, 47, 16, and 0% of the samples were seropositive for at least one Leptospira serovar, CDV, and PRRSV, respectively. Most of the positive Leptospira results were to serovars grippotyphosa (36%), autumnalis (22%), and hardjo (22%). No statistically significant correlation was found between antibody prevalence estimates for different Leptospira serovars. A significant association was found between body weight and antibodies for Leptospira serovars and CDV. In addition, age (adult vs. juvenile) was significantly associated with the presence of CDV antibody, with adults exhibiting a higher prevalence than juveniles. This study confirmed that raccoons in Indiana, USA, are exposed to different Leptospira interrogans serovars and CDV and that age and weight are associated with the presence of antibodies for both pathogens.

The use of harvested white-tailed deer (Odocoileus virginianus) and geographic information system (GIS) methods to characterize distribution and locate spatial clusters of Borrelia burgdorferi and its vector Ixodes scapularis in Indiana.

Ixodes scapularis (Say) is the vector for Borrelia burgdorferi (Bb) the causative agent of Lyme disease (LD). The increased number and presence of ticks in the environment pose a significant health risk to people and many domestic animals including dogs, cats, and horses. This study characterized the distribution and expansion of I. scapularis and Bb and identified areas of increased risk of LD transmission in Indiana using geographical information systems (GIS) and spatial analysis. A cross-sectional sampling was performed for 3 consecutive years (2005-2007). A total of 3,412 harvested white-tailed deer (Odocoileus virginianus) were searched for ticks at Department of Natural Resources manned deer check-in stations. Hunters were asked for verbal permission to search the deer and to indicate on a road atlas where the deer was killed. All deer points were digitized into a GIS database. Identification of clustering in space and time for these organisms was performed using geostatistical software. Multiple spatial clusters of I. scapularis-infested deer were identified in western Indiana. B. burgdorferi was isolated from tick pools in 11 counties. In addition to the I. scapularis clusters, one spatial cluster of Bb-infected ticks was identified. Our current survey results and cluster analysis indicate that the western geographic regions of Indiana should be considered by the healthcare community to be at increased risk of LD compared with the rest of Indiana.

Catheter related bacteremia in rats: A preliminary report on the effect of methylene blue coated catheter.

The safety and efficacy of methylene blue (MB) coated indwelling jugular vein/cranial vena cava catheter made up of polyurethane material was tested in a rat model, receiving bacterial culture suspension of Pseudomonas aeruginosa, and Staphylococcus aureus. Daily blood samples were collected from the catheter and peripheral vein for bacterial culture. The clinical parameters (rectal temperature, respiratory rate, total white blood cell count, and loss in body weight) were not different between the groups. All the rats became bacteremic with similar changes in the number of colony forming units in the catheter and peripheral samples. Histopathological lesions were not different between the groups. The findings suggest that rats receiving MB coated catheters behaved similar to non-coated catheters. Based on the results it can be concluded that for this type of gross contamination, catheter coating alone may not eliminate infection/bacteremia.

Characterization of turkey coronavirus from turkey poults with acute enteritis.

The present study was to characterize turkey coronavirus associated with turkey poult enteritis and mortality. Intestinal contents or intestines from affected turkey poults and inoculated turkey embryos contained coronaviruses as revealed by electron microscopy or were positive for turkey coronavirus by immunofluorescent antibody assay. Sucrose density gradient ultracentrifugation of the virus-containing intestinal homogenate yielded two opalescent bands corresponding to the buoyant densities of 1.14-1.15 and 1.18-1.20 g/ml, respectively. Coronaviral particles from intestinal contents or the sucrose density gradient preparation were mainly spherical in shape and had envelope and central depression. They were surrounded by a fringe of regularly spaced petal-shaped projections attached to the particles by a short stalk. Purified viruses hemagglutinated rabbit erythrocytes with a titer of 16. Major protein bands of purified viruses analyzed by SDS-PAGE were located at 200, 100-110, 50-60, and 30-35 kDa. The patterns of protein bands were consistent with those of Minnesota or Quebec turkey coronavirus isolates. A 568 bp nucleotide fragment of turkey coronavirus spike protein gene was amplified from RNA of inoculated turkey embryo intestine or purified virus. Sequence analysis of the 568 bp PCR product revealed high degree of identity with the corresponding spike protein gene sequence of human and bovine coronaviruses. The results indicated that turkey coronavirus was associated with turkey poults with acute enteritis.