Migration of retrobulbar wooden foreign body between diagnostic imaging and surgical extraction in a German shepherd dog

A 2‐year‐old, male castrated German shepherd dog was presented to the University of Tennessee Veterinary Medical Center (UTVMC) with periorbital swelling and conjunctival mucopurulent discharge 2 days following removal of a twig from the medial canthus by the owner. Diagnostic imaging was pursued due to the suspicion of a retrobulbar foreign body (FB). A cylindrical FB approximately 3.0 cm in length and 1.0 cm in diameter with concentric rings, suspected to be wooden material, was identified on computed tomography (CT) imaging. An attempt to remove the FB via a stab incision using ultrasound guidance was unsuccessful, and postmanipulation ultrasound confirmed the FB position was unchanged. An exploratory orbitotomy was performed, using the acquired CT images for guidance in locating the FB; however, the FB was not present at the predicted site. The CT imaging was repeated and showed that the FB had migrated rostrally approximately 3.0 cm, compared to the originally acquired study and its same location during attempted ultrasound‐guided removal. A combination of CT‐guided needle placement and contrast injection was then used with repeat imaging in an attempt to better localize the FB and its soft tissue tract. The dog was taken back into the operating room, and the wooden FB was successfully removed.     

Cumulative racing-speed exercise distance cluster as a risk factor for fatal musculoskeletal injury in Thoroughbred racehorses in California

Thoroughbred racehorses which suffered a fatal musculoskeletal injury (FMI) while racing or race training at a California racetrack during 9 months of 1991 were studied to determine the importance of intensive, high-speed exercise schedules prior to injury. Seventy-seven horses which sustained an FMI while racing and 45 horses which sustained an FMI while race training were successfully matched by race or timed workout session with one control horse and included in the analyses. Race and timed workout (racing-speed exercise) histories were obtained for the case and control horses. Two-month cumulative, racing-speed cutoff distances were calculated from the control horse sample by two methods. Median racing-speed exercise frequencies and distances of the control horses were used to estimate age-specific (2, 3, 4 and ≥ 5 years), 2-month cumulative, racing-speed distances (Method 1). For the second method, the last race or timed workout for each control horse occurring just prior to, or on the date of injury for the matched case horse was identified. Cumulative racing-speed distances 2 months prior to these exercise events were determined for each control horse and used to estimate median age-specific (2, 3, 4 and ≥ 5 years), 2-month cumulative racing-speed distances (Method 2). The cumulative cutoff distances estimated from both methods were used to classify each matched pair according to the presence or absence of a 2-month cumulative, racing-speed distance which exceeded the age-appropriate cutoff distance (exercise distance cluster) within 6 months prior to injury. Manlel-Haenszel matched-pair odds ratios and 95% confidence limits were calculated separately for the racing and race-training fatal injuries. The relative risk for racing FMI was significantly greater for those horses which ran 2-month, cumulative racing and timed workout distances in excess of the cutoff values determined with Method 1 (relative risk (RR) = 3.0, 95% confidence interval (CI) = 1.2, 7.6) and Method 2 (RR = 7.2, 95% CI = 2.6, 20.6). The relative risk for race-training FMI was significantly greater for those horses which ran 2-month, cumulative racing and timed workout distances in excess of the cutoff values determined with Method 2 (RR = 3.4, 95% CI =1.0, 13.2).     

Detection of fluoroquinolone resistance level in clinical canine and feline Escherichia coli pathogens using rapid real-time PCR assay

Fluoroquinolones are used to treat infections caused by Escherichia coli in canine and feline veterinary patients, particularly those infecting the urinary tract. The gyrA gene is a primary target causing fluoroquinolone resistance in Gram negative coliforms, with mutations in codons 83 and 87 generally associated with high-level of resistance E. coli clinical isolates. We have developed a fluorescence resonance energy transfer (FRET) quantitative PCR to identify enrofloxacin-resistance in clinical E. coli isolates that carry mutations in codons 83 and 87 of gyrA. This real-time quantitative PCR assay is rapid, economical, and sensitive compared with cultured antimicrobial susceptibility testing. The assay identified as few as four genome copies per reaction from culture and 19 genome copies in urine. For the 70 isolates tested, the sensitivity was 87.5% (95% CI = 75–95.3%) (n = 42/48), specificity was 100% (95% CI = 87.3–100%) (n = 22/22), whereas accuracy was 91.4% (95% CI = 82.3–97%) (n = 64/70). Furthermore, we were able to accurately differentiate between the wild type and mutants E. coli directly from infected canine urine samples (n = 5) within 2 h. These results were confirmed by sequence alignments of the PCR products and comparison with the susceptibility testing. The FRET-PCR assay appears to have promising clinical application as an early diagnostic tool for rapid and sensitive detection and differentiation of the level of fluoroquinolone resistance among clinical E. coli isolates that may facilitate design of the dosing regimen.     

Bovine NK cells acquire cytotoxic activity and produce IFN-gamma after stimulation by Mycobacterium bovis BCG- or Babesia bovis-exposed splenic dendritic cells

Early interactions of innate immune cell populations, such as dendritic cells (DC) and natural killer (NK) cells, can affect the ability of the acquired immune response to control infection of intracellular microorganisms. In this study, we investigated the activation of bovine NK cells by CD13(+) splenic DC stimulated with either Mycobacterium bovis BCG or Babesia bovis merozoites. Splenic DC were used either immediately after selection (cytokine(-)) or after exposure to GM-CSF, IL-4 and Flt3L for 72 h (cytokine(+)). Phenotypic analyses showed up-regulation of MHCII, CD80 and CD86 on cytokine(+) DC when compared to cytokine(-) DC. Purified NK cells (CD335(+)CD3(-)CD2(+/-)CD8alpha(+/-)) were co-cultured with microbial-exposed cytokine(-) DC or cytokine(+) DC in either transwell or cell-to-cell format and NK cell IFN-gamma production and cytotoxicity were assessed. NK cell IFN-gamma production was dependent on cell-to-cell contact. Microbial-stimulated cytokine(+) DC induced significantly more IFN-gamma production from NK cells than cytokine(-) cells. In contrast, cytotoxicity and perforin up-regulation were more pronounced in NK cells cultured with cytokine(-) DC than cytokine(+) DC. Therefore, activation of bovine NK cells by microbial-stimulated CD13(+) splenic DC is influenced by the maturation state of the DC suggesting different roles for the splenic DC during disease-induced maturation.     

Absence of three known benzimidazole resistance mutations in Trichostrongylus tenuis, a nematode parasite of avian hosts

Benzimidazole (BZ) resistance is widespread in nematode parasites of livestock, but very little is known about the levels of BZ resistance in parasites with avian hosts. We investigated BZ resistance in Trichostrongylus tenuis, a nematode parasite of red grouse, Lagopus lagopus scotica. BZ anthelmintics had been in use in this system for up to 15 years, yet existing phenotypic evidence for resistance was inconclusive. We screened 1530 individuals from 14 populations at the principal β-tubulin locus for BZ resistance (isotype 1, residue 200) and 940 of these at two further resistance sites (isotype 1, residue 167; isotype 2, residue 200). No BZ resistant genotypes were found. Alternative mechanisms may be responsible for BZ resistance in this system, or the method and timing of treatments may reduce selection pressure for BZ resistance by creating substantial refugia for susceptible genotypes.