Bacterial-associated diarrhea in the dog: a critical appraisal.

The clinical documentation of enteropathogenic bacteria causing diarrhea in dogs is clouded by the presence of many of these organisms existing as normal constituents of the indigenous intestinal flora. The diagnosis of a putative bacterial enteropathogen(s) in dogs should be made based on a combination of parameters, including signalment and predisposing factors, clinical signs, serologic assays for toxins, fecal culture, and PCR. Relying on results of fecal culture alone is problematic, because C perfringens, C difficile, Campylobacter spp, and pathogenic and non-pathogenic E coli are commonly isolated from apparently healthy dogs [10,13,33]. Nevertheless, culture may be useful in procuring isolates for the application of molecular techniques, such as PCR, for detection of specific toxin genes or molecular typing of isolated strains to establish clonality in suspected outbreaks. The oversimplistic attempt to characterize bacterially associated diarrhea by anatomic localization of clinical signs should be discouraged, because most of the previously mentioned bacteria have been associated with small and large intestinal diarrhea. Accurate diagnosis of infections may require diagnostic laboratories to incorporate PCR-based assays using genus- and species-specific primers to facilitate detection of toxin genes and differentiation of species that appear phenotypically and biochemically similar. There has been tremendous interest in the application of microarray technology for the simultaneous detection of thousands of genes or target DNA sequences on one glass slide. This powerful tool could be used for detection of specific pathogenic bacterial strains in fecal specimens obtained from dogs in the future.     

Validation of the 13C-urea breath test for use in cheetahs (Acinonyx jubatus) with Helicobacter.

Historically, therapeutic monitoring for prescribed eradication treatment of Helicobacter in cheetahs (Acinonyx jubatus) with associated gastritis has been accomplished only through endoscopic biopsies. The 13C-urea breath test (UBT) can offer an alternative to repeated biopsies for therapeutic monitoring. Five male and five female cheetahs and one male Sumatran tiger (Panthera tigris) were studied. All were clinically healthy before and after this investigation. Breath samples of end-tidal expiration were taken before and after administration of a 13C-enriched urea solution through a gastroesophageal tube. Twenty-milliliter breath samples were taken at 10, 20, 30, and 40 min after administration of the urea solution. The results of the breath analysis were compared with the results of rapid urease testing, histopathologic examination, and impression smears of gastric biopsies taken at the time of the breath test. The sensitivity and specificity for the 13C-UBT in this investigation were 100%. and the positive predictive value and negative predictive value were both 100%. Although the 13C-UBT is a good noninvasive diagnostic tool for monitoring the presence of Helicobacter sp. in the gastric mucosa, endoscopy should still be used for initial diagnosis and grading of gastritis and for monitoring the progression of disease in cheetahs. The 13C-UBT is a valuable, simple, accurate, and sensitive tool for monitoring eradication of Helicobacter during therapy for clinical gastritis.     

Trichoderma Biocontrol of Colletotrichum acutatum and Botrytis cinerea and Survival in Strawberry

Trichoderma isolates are known for their ability to control plant pathogens. It has been shown that various isolates of Trichoderma, including T. harzianum isolate T-39 from the commercial biological control product TRICHODEX, were effective in controlling anthracnose (Colletotrichum acutatum) and grey mould (Botrytis cinerea) in strawberry, under controlled and greenhouse conditions. Three selected Trichoderma strains, namely T-39, T-161 and T-166, were evaluated in large-scale experiments using different timing application and dosage rates for reduction of strawberry anthracnose and grey mould. All possible combinations of single, double or triple mixtures of Trichoderma strains, applied at 0.4% and 0.8% concentrations, and at 7 or 10 day intervals, resulted in reduction of anthracnose severity; the higher concentration (0.8%) was superior in control whether used with single isolates or as a result of combined application of two isolates, each at 0.4%. Only a few treatments resulted in significant control of grey mould. Isolates T-39 applied at 0.4% at 2 day intervals, T-166 at 0.4%, or T-161 combined with T-39 at 0.4% were as effective as the chemical fungicide fenhexamide. The survival dynamics of populations of the Trichoderma isolates (T-39, T-105, T-161 and T-166) applied separately was determined by dilution plating and isolates in the mixtures calculated according to the polymerase chain reaction (PCR) using repeat motif primers. The biocontrol isolates were identified to the respective species T. harzianum (T-39), T. hamatum (T-105), T. atroviride (T-161) and T. longibrachiatum (T-166), according to internal transcribed spacer sequence analysis.     

Does sand content in spawning substrate result in early larval emergence? Evidence from a lithophilic cyprinid fish

The spawning success of lithophilic salmonids is strongly influenced by the fine sediment content (“fines”) of spawning substrates, yet knowledge on the impacts of fines on the spawning of non‐salmonid lithophiles remains limited, despite their ecological and socio‐economic importance in European rivers. Consequently, the aim here was to use an ex‐situ experiment to investigate the impact of sand content on egg survival and timing of larval emergence of the surface‐spawning cyprinid European barbel Barbus barbus. Thirty incubator boxes within a recirculating system were filled with one of five experimental sediment mixtures (0%–40% sand by mass) that each contained 300 fertilised eggs at a depth of 50 mm. Emerged, free‐swimming larvae were captured and counted daily to assess grain‐size effects on larval survival and emergence. Specifically, total proportion of emerged larvae, cumulative daily proportion of emerged larvae and time required to reach 50% emergence were measured during the study. Whilst the proportion of sand in the sediments did not have a significant impact on egg‐to‐emergence survival (mean survival per treatment 75%–79%), it significantly affected the timing of larval emergence to the water column; early emergence was detected in treatments with elevated sand content (on average, 50% emergence after 12–13 days versus 19 days in the control). Similar to findings from salmonid studies, these results suggest high sand content in spawning gravels can influence timing of larval emergence and potentially cyprinid lithophilic fish survival.     

Elimination of cetirizine following administration of multiple doses to exercised thoroughbred horses

Cetirizine is an antihistamine used in performance horses for the treatment of hypersensitivity reactions and as such a withdrawal time is necessary prior to competition. The objective of the current study was to describe the disposition and elimination of cetirizine following oral administration in order to provide additional serum concentration data upon which appropriate regulatory recommendations can be established. Nine exercised thoroughbred horses were administered 0.4 mg/kg of cetirizine orally BID for a total of five doses. Blood samples were collected immediately prior to drug administration and at various times postadministration. Serum cetirizine concentrations were determined and selected pharmacokinetic parameters determined. The serum elimination half‐life was 5.83 ± 0.841 h. Average serum cetirizine concentrations were still above the LOQ of the assay (0.05 ng/mL) at 48 h (final sample collected) postadministration of the final dose.     

Pharmacokinetics of methocarbamol and phenylbutazone in exercised Thoroughbred horses

Methocarbamol (MCBL) is commonly used in performance horses for the treatment of skeletal muscle disorders. Current regulatory recommendations for show horses and racehorses are based on a single oral dose of 5 g, although doses in excess of this are often administered. The goal of the current study was to characterize the disposition of MCBL following higher dose administration and administration in combination with another commonly used drug in performance horses, phenylbutazone (PBZ). Exercised Thoroughbred horses were administered various doses of MCBL as a sole agent and MCBL in combination with PBZ. Blood samples were collected at various times, concentrations of MCBL and PBZ measured using LC‐MS/MS and pharmacokinetic parameters calculated using compartmental analysis. Following administration of 15 g of MCBL, either as part of a single‐ or multiple‐dose regimen, a number of horses exceeded the Association of Racing Commissioners International and the United States Equestrian Federation's recommended regulatory threshold at the recommended withdrawal time. There was not a significant difference between horses that received only MCBL and those that received MCBL and PBZ. Results of the current study support an extended withdrawal guideline when doses in excess of 5 g are administered.     

Sialidase activity in Mycoplasma synoviae

Eleven strains of the avian pathogen Mycoplasma synoviae were evaluated for the presence of sialidase activity with the use of the fluorogenic substrate 2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid and the sialidase inhibitor 2-deoxy-2,3- didehydro-N-acetylneuraminic acid. The kinetics of in vitro growth in modified Frey medium were also assessed for each strain. Five strains had been isolated from clinically symptomatic chickens, and strains WVU 1853T and K3344 have been demonstrated to be capable of reproducing disease in specific-pathogen-free chickens. All strains exhibited sialidase activity, although the amount varied 65-fold among strains (P < 0.0001) from 1.3 x 10(-7) to 2.0 x 10(-9) activity units per colony-forming unit. Strains originally isolated from clinically symptomatic birds had more (P < 0.05) sialidase activity than strains from asymptomatic birds. Strain WVU1853T exhibited the most sialidase activity (P < 0.0001) and grew to the highest culture density (P < 0.0001) among strains, but across strains, the rank correlation of growth rate with sialidase activity was not significant. Negligible activity was detected in conditioned culture supernatant fluid. This is the first report of sialidase activity in pathogenic strains of M. synoviae, which suggests a potential enzymatic basis for virulence of the organism.