Conventional direct microscopy with potassium hydroxide (KOH) and culture were found to lack the ability to establish a fast and specific diagnosis of dermatophytosis. A pan-dermatophyte nested-PCR assay was developed using a novel primer pair targeting the translation elongation factor 1-α (Tef-1α) sequences for direct detection and identification of most veterinary relevant dermatophytes in animal samples suspected to dermatophytosis. A total of 140 animal skin and hair samples were subjected to direct microscopy, culture, and ITS-RFLP/ITS-sequencing of culture isolates for the detection and identification of dermatophytosis agents. Nested-PCR sequencing was performed on all the extracted DNAs using a commercial kit after dissolving the specimens by mechanical beating. Nested-PCR was positive in 90% of samples, followed by direct microscopy (85.7%) and culture (75%). The degree of agreement between nested-PCR and direct microscopy (94.4%) was higher than with culture (83.3%). In 105 culture-positive cases, the measures of agreement for the identification of dermatophytosis agents were as follows: 100% between nested-PCR sequencing and ITS-RFLP/ITS-sequencing and 63.8% between nested-PCR sequencing and culture. The developed nested-PCR was faster as well as more sensitive and specific than conventional methods for detection and identification of dermatophytes in clinical samples, which was particularly suitable for epidemiological studies.
A 4-year-old Iranian boy developed erythematous, itchy and annular lesion on his face. Microscopic examination of the scraped samples with 10 % potassium hydroxide (KOH) revealed fungal septate hyphae and arthroconidia. The etiological agent was found to be Microsporum gypseum in mycological examinations. Amplification and restriction digestion of the internal transcribed spacers (ITS) of rDNA was not helpful for identification, but in ITS sequencing the isolate showed 98 % homology to Microsporum incurvatum strain CBS 172.64. Empirical treatment of the patient with griseofulvin for 4 weeks was successful. Other than our isolate, the ITS1 sequences of 38 strains from related species were retrieved from GenBank and phylogenetic tree using maximum likelihood method was constructed. The case isolate clustered apart from other strains of M. incurvatum. Pairwise comparison of ITS1 showed intraspecies variations of 0-13 nucleotides among M. incurvatum strains and an extensive interspecies variation of 33-80 bp and remarkable interspecies size polymorphism between the three sister species in the M. gypseum complex. The high level of ITS1 intraspecific variation is suitable for species identification rather than phylogeographic analysis of M. gypseum complex.
The objective of the present study was to compare the effects of propofol and ketofol on intraocular pressure, tear production and cardiorespiratory variables in dogs premedicated with midazolam. Six castrated adult mixed-breed dogs were used in a cross-over design with a one-week interval. Twenty minutes after premedication with midazolam (0.2 mg/kg), animals were assigned randomly to two groups and received either propofol (6 mg/kg) or ketofol (3 mg/kg; 1 : 1 mg/ml ratio) treatments intravenously. Intraocular pressure, tear production, heart rate, respiratory rate, rectal temperature and direct mean arterial blood pressure were measured at base (before induction), and at 5, 10, 15, 20 and 30 min after induction of anaesthesia. Blood gas samples were obtained at base (before induction), and at 5, 15 and 30 min after administration of treatments. Intraocular pressure showed significantly higher values at 5 min after induction in ketofol compared with propofol (16.1 ± 4.5 mm Hg vs 8.2 ± 1.2 mm Hg, respectively). There were no significant changes in tear production in either group. Significantly higher heart rate and mean arterial blood pressure were detected in ketofol at several time points. Respiratory depression occurred in both groups with no significant differences between them. In conclusion, although ketofol improved heart rate and mean arterial blood pressure and did not elicit more pronounced respiratory depression than propofol, it resulted in significantly higher values of intraocular pressure at 5 min after administration in dogs. Despite the small number of dogs in this study, our results indicate that ketofol should not be recommended for ophthalmic surgical procedures in dogs. Appropriate oxygenation should be provided when propofol is used for ophthalmic surgeries.
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