Analysis of the effects of storage temperature and contamination on aerobic bacterial culture results of bronchoalveolar lavage fluid.

BACKGROUND: Storage temperature of bronchoalveolar lavage fluid (BALF) impacts cytological evaluation. The effect of storage temperature before bacterial culture has not been evaluated. OBJECTIVES: To assess whether BALF storage temperature alters aerobic bacterial culture results. ANIMALS: Eight healthy, male, intact, purpose-bred Beagles. METHODS: Prospective, controlled investigation. Samples of BALF were collected sterilely. Half of each sample was reserved for controls, and half was inoculated with 104 colony forming units per milliliter (cfu/mL) Bordetella bronchiseptica and 102 cfu/mL Escherichia coli. Control and inoculated samples each were separated into 4 aliquots (1 plated immediately; 3 stored at 4, 24, or 37°C, respectively, for 24 hours before aerobic bacterial culture). Colony counts were compared across treatments for each organism. RESULTS: In inoculated samples, a statistical difference could not be detected in growth of E. coli or B. bronchiseptica between the baseline culture and BALF stored at 4°C for 24 hours before culture. However, for E. coli, growth in cfu/mL at both 24 and 37°C was higher compared to baseline (P < .05) and compared to 4°C (P < .05). For B. bronchiseptica cfu/mL, growth at 37°C was significantly different (P = .003) compared to both baseline and 4°C. CONCLUSIONS AND CLINICAL IMPORTANCE: Samples of BALF may be stored at 4°C for 24 hours before culture without substantially altering culture results. Inappropriate storage or shipment temperature (room temperature or exposure to heat) can result in overgrowth of E. coli or B. bronchiseptica, which could alter clinical decisions.

Minimum inhibitory concentration and killing properties of rifampicin against canine Staphylococcus pseudintermedius isolates from dogs in the southeast USA.

BACKGROUND: Meticillin-resistant (MR) staphylococcal pyoderma in dogs has led to increased use of alternate antibiotics such as rifampicin (RFP). However, little information exists regarding its pharmacodynamics in MR Staphylococcus pseudintermedius. HYPOTHESIS/OBJECTIVES: To determine the minimum inhibitory concentration (MIC) and killing properties of RFP for canine Staphylococcus pseudintermedius isolates. METHODS: The MIC of RFP was determined using the ETEST® for 50 meticillin-susceptible (MS) and 50 MR S. pseudintermedius isolates collected from dogs. From these isolates, two MS isolates (RFP MIC of 0.003 and 0.008 µg/mL, respectively) and two MR isolates (RFP MIC of 0.003 and 0.012 µg/mL, respectively) were subjected to time-kill studies. Mueller-Hinton broth was supplemented with RFP at 0, 0.5, 1, 2, 4, 8, 16 and 32 times the MIC for 0, 2, 4, 10, 16 and 24 h. The number of viable colony forming units in each sample was determined using a commercial luciferase assay kit. RESULTS: The MIC50 and MIC90 were the same for MS and MR isolates, at 0.004 µg/mL and 0.008 µg/mL, respectively. Rifampicin kill curves were not indicative of concentration-dependency, suggesting time-dependent activity. Two isolates (MS 0.003 and 0.008 µg/mL) exhibited bacteriostatic activity, whereas two others (MR 0.003 and 0.012 µg/mL) exhibited bactericidal activity. CONCLUSIONS AND CLINICAL IMPORTANCE: This study demonstrated that MS and MR S. pseudintermedius isolates were equally susceptible to rifampicin and that dosing intervals should be designed for time-dependent efficacy. These data can support pharmacokinetic studies of RFP in dogs with susceptible infections caused by S. pseudintermedius.

Presurgical antiseptic efficacy of chlorhexidine diacetate and providone-iodine in the canine preputial cavity.

Antiseptic flushing of the canine prepuce and its exclusion from the surgical field are recommended before abdominal surgery to reduce the risk of bacterial contamination. The authors cultured the preputial cavity of 60 dogs prior to and following flushing with 0.05% chlorhexidine diacetate, 1% povidone-iodine, or 0.9% saline control. Bacterial growth was evaluated using a semiquantitative method, and bacterial organisms were subsequently identified. There were no significant differences between povidone-iodine and the saline control in any of the variables assessed. Chlorhexidine resulted in a significant decrease in the proportion of positive postflush cultures compared with povidone-iodine. Although not significant, the difference in adverse reactions between povidone-iodine (25%) and chlorhexidine diacetate (5%) suggests clinical relevance. Based on the results of this study, a 2 min flush with 0.05% chlorhexidine diacetate is recommended for presurgical preparation of the preputial cavity.

An inversion of 25 base pairs causes feline GM2 gangliosidosis variant.

In G(M2) gangliosidosis variant 0, a defect in the beta-subunit of lysosomal beta-N-acetylhexosaminidase (EC 3.2.1.52) causes abnormal accumulation of G(M2) ganglioside and severe neurodegeneration. Distinct feline models of G(M2) gangliosidosis variant 0 have been described in both domestic shorthair and Korat cats. In this study, we determined that the causative mutation of G(M2) gangliosidosis in the domestic shorthair cat is a 25-base-pair inversion at the extreme 3' end of the beta-subunit (HEXB) coding sequence, which introduces three amino acid substitutions at the carboxyl terminus of the protein and a translational stop that is eight amino acids premature. Cats homozygous for the 25-base-pair inversion express levels of beta-subunit mRNA approximately 190% of normal and protein levels only 10-20% of normal. Because the 25-base-pair inversion is similar to mutations in the terminal exon of human HEXB, the domestic shorthair cat should serve as an appropriate model to study the molecular pathogenesis of human G(M2) gangliosidosis variant 0 (Sandhoff disease).

Effects of 3 biologic dressings on healing of cutaneous wounds on the limbs of horses.

Three biologic dressings [split-thickness allogeneic skin (STS)], allogeneic peritoneum (P), and xenogenic porcine small intestinal submucosa (PSIS)] were studied to determine their effects on bacterial proliferation, inflammatory reaction, vascularization, and overall healing and to compare the effects of these dressings with the effects of a nonbiologic dressing, a nonadherent synthetic pad (NASP). A medial wound (3 cm in diameter) and 2 lateral wounds (2 cm in diameter) were created at the junction of the proximal and middle thirds of each metacarpus and metatarsus in 5 horses. Each medial wound and the proximolateral wound received an STS, P, PSIS, or NASP dressing on day 8 after wounding. The other lateral wound received an NASP dressing. Bacterial proliferation, inflammatory reaction (histologic changes), and drhessing vascularization were evaluated 6 d after application of the dressing. Percentages of contraction and epithelialization, as well as healing time, were determined when the wounds had completely epithelialized. The practical applicability of the different dressings to equine wound management was also assessed. No significant difference was detected in the parameters evaluated among the treated wounds or between the treated and control wounds. The biologic dressings had no effect on infection, inflammatory response, or healing time. Vascularization was not identified in any of the biologic dressings. The PSIS and P dressings required numerous applications over the study period. The STS dressings are more practical than PSIS and P dressings owing to ease of application and stability. Thus, these biologic dressings offer no apparent advantage over a nonbiologic dressing for treatment of small granulating wounds.

Isolation and characterization of multipotential mesenchymal stem cells from feline bone marrow.

OBJECTIVE: Although several types of stem cells have been isolated from rodent and human tissues, very few data exist on stem cell isolation from nonrodent animals, which seriously limits the advancement of stem cell biology and its ultimate translation to human clinical applications. Domestic cats are used frequently in biomedical research and are the preferred species for studies of normal physiology and disease, particularly in neuroscience. Therefore, the objective of this study was to characterize mesenchymal stem cells (MSC) from feline bone marrow for use in research on the application of stem cells to human health problems for which cats are the preferred model. METHODS: Mesenchymal stem cells from feline bone marrow were isolated by standard methodology developed for other species and characterized according to morphology, growth traits, cell-surface antigen profile, and differentiation repertoire in vitro. RESULTS: Feline mesenchymal stem cells exhibit a fibroblast-like morphology with bipolar or polygonal cell bodies and possess a cell-surface antigen profile similar to their rodent and human counterparts. Feline MSC exist at a frequency of 1 in 3.8 x 10(5) bone marrow mononuclear cells and are capable of differentiation to adipocytic, osteocytic, and neuronal phenotypes when exposed to appropriate induction media. CONCLUSIONS: Mesenchymal stem cells isolated from feline bone marrow possess several traits typical of MSC from other species. Characterization of feline mesenchymal stem cells will facilitate future studies of stem cell biology and therapeutics for which the domestic cat is an indispensable model.