Chromogenic culture media complements diagnostic cytology in the visual identification of pathogenic skin bacteria in dogs and cats.

In dogs and cats, bacterial skin infections (pyoderma and otitis externa) are a common cause for visiting the veterinary clinic. The most frequent skin pathogens are Staphylococcus pseudintermedius, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, often requiring different therapeutic antibiotic protocols. Unfavorably, existing diagnostics based on cytology cannot reveal bacterial species but only bacterial shapes such as cocci or rods. This microscopic limitation could be overcome by clinical translation of affordable chromogenic media, which enable species identification based on bacterial colonies growing in different colors and sizes. In this study, we determined how well inexperienced general veterinary clinicians identified bacterial pathogens from the skin and ears on two commercial (Chromatic™ MH and Flexicult® Vet) and one custom-made Mueller Hinton agar-based chromogenic medium. For this purpose, four veterinarians evaluated 100 unique samples representing 10 bacterial species. On average, clinicians correctly identified between 72.1 and 86.3% of bacterial species. Colony colors developed quickly on the Chromatic™ MH medium, leading to the highest 81.6% identification accuracy after 24 h incubation. However, Flexicult® Vet exhibited the highest accuracy of 86.3% after prolonged 48 h incubation. Evaluators easily recognized bacteria displaying uniquely colored colonies like green-brown Pseudomonas aeruginosa, blue Enterococcus faecalis, orange-brown Proteus spp., and red Escherichia coli. Oppositely, staphylococci shared uncharacteristically pale pink colonies causing misidentifications among the genus, deteriorating overall accuracy by around 10 percentage points (from 90.9%). Another reason for identification errors was the evaluators' inexperience, reflected in not recognizing colony size differences. For example, although Streptococcus canis exhibited the tiniest colonies, the species was frequently mistaken for other cocci. Finally, around 10% of errors were negligence-related slips due to unconsidered sample history. To conclude, the introduction of chromogenic media into veterinary clinics can significantly complement diagnostics in skin inflammations by identifying pathogen species in around 80% of cases. The extra information may help in therapeutic dilemmas on antibiotics and standard antimicrobial susceptibility testing. Additional personnel training and evaluation help by visuals, flowcharts, checklists, and, if necessary, microbiologists could further improve identification accuracy.

Multimodal Approach of Optical Coherence Tomography and Raman Spectroscopy Can Improve Differentiating Benign and Malignant Skin Tumors in Animal Patients.

As in humans, cancer is one of the leading causes of companion animal mortality. Up to 30% of all canine and feline neoplasms appear on the skin or directly under it. There are only a few available studies that have investigated pet tumors by biophotonics techniques. In this study, we acquired 1115 optical coherence tomography (OCT) images of canine and feline skin, lipomas, soft tissue sarcomas, and mast cell tumors ex vivo, which were subsequently used for automated machine vision analysis. The OCT images were analyzed using a scanning window with a size of 53 × 53 µm. The distributions of the standard deviation, mean, range, and coefficient of variation values were acquired for each image. These distributions were characterized by their mean, standard deviation, and median values, resulting in 12 parameters in total. Additionally, 1002 Raman spectral measurements were made on the same samples, and features were generated by integrating the intensity of the most prominent peaks. Linear discriminant analysis (LDA) was used for sample classification, and sensitivities/specificities were acquired by leave-one-out cross-validation. Three datasets were analyzed-OCT, Raman, and combined. The combined OCT and Raman data enabled the best sample differentiation with the sensitivities of 0.968, 1, and 0.939 and specificities of 0.956, 1, and 0.977 for skin, lipomas, and malignant tumors, respectively. Based on these results, we concluded that the proposed multimodal approach, combining Raman and OCT data, can accurately distinguish between malignant and benign tissues.

Evaluation of native canine skin color by smartphone-based dermatoscopy.

BACKGROUND: Human skin color, predominantly determined by the chromophores of melanin, hemoglobin, and exogenous carotenoids, is often measured to serve various medical and cosmetic applications. Although colorimetry has been used to evaluate the skin erythema in allergic dogs, the native canine skin color remains unknown. METHODS: We measured the skin color in 101 healthy dogs using a calibrated optical system with a smartphone and a mobile dermatoscope DermLite DL1. The results were retrieved in the CIELAB color system and compared to the human color ranges. RESULTS: The lightness (L*) of canine skin ranged from 28.5 to 78.3, which is slightly broader than that of human skin. There was a difference of 3.9 in redness (a*) between canine and human skin, but this variation could be attributed to the similarly valued colorimetric error of the optical system. Nonetheless, the skin yellowness was significantly different for dogs and humans (respective median b* of 12.3 versus 16.6, p < 0.01). This difference might be due to canids not being able to accumulate typically yellowish carotenoids. Furthermore, the native canine skin color did not exhibit a typical dependence between the coordinates of lightness (L*) and yellowness (b*), known as the individual typology angle, °ITA. CONCLUSION: We reported the first dataset of the native canine skin color in the CIELAB color space. We discovered a similarity in skin lightness and a difference in skin yellowness. However, further studies are needed for a more precise comparison of skin redness.

How Accurate Are Veterinary Clinicians Employing Flexicult Vet for Identification and Antimicrobial Susceptibility Testing of Urinary Bacteria?

Antibiotics are frequently used for treating urinary tract infections (UTI) in dogs and cats. UTI often requires time-consuming and expensive antimicrobial susceptibility testing (AST). Alternatively, clinicians can employ Flexicult Vet, an affordable chromogenic agar with added antibiotics for in-clinic AST. We investigated how well veterinary microbiologists and clinicians, without any prior experience, employ Flexicult Vet for the identification and AST of the most common canine and feline urinary pathogenic bacteria. We prepared 47 monoculture plates containing 10 bacterial species. The test's mean accuracy was 75.1% for bacteria identification (84.6% and 68.7% for microbiologists and clinicians, respectively) and 79.2% for AST (80.7% and 78.2%). All evaluators employed Flexicult Vet with the accuracies over 90% for the distinctively colored bacteria like Escherichia coli (red), Enterococcus faecalis (turquoise), and Proteus spp. (pale brown). However, the evaluators' experience proved important in recognizing lightly colored bacteria like Staphylococcus pseudintermedius (accuracies of 82.6% and 40.3%). Misidentifications of E. faecium additionally worsened AST performance since bacterial intrinsic resistance could not be considered. Finally, only 33.3% (3/9) of methicillin-resistant S. pseudintermedius (MRSP) were correctly detected. To conclude, Flexicult Vet proved reliable for certain urinary pathogens. In contrast, light-colored bacteria (e.g., Staphylococcus), often misidentified, require a standard AST.

Evaluation of Erythema Severity in Dermatoscopic Images of Canine Skin: Erythema Index Assessment and Image Sampling Reliability.

The regular monitoring of erythema, one of the most important skin lesions in atopic (allergic) dogs, is essential for successful anti-allergic therapy. The smartphone-based dermatoscopy enables a convenient way to acquire quality images of erythematous skin. However, the image sampling to evaluate erythema severity is still done manually, introducing result variability. In this study, we investigated the correlation between the most popular erythema indices (EIs) and dermatologists' erythema perception, and we measured intra- and inter-rater variability of the currently-used manual image-sampling methods (ISMs). We showed that the EIBRG, based on all three RGB (red, green, and blue) channels, performed the best with an average Spearman coefficient of 0.75 and a typical absolute disagreement of less than 14% with the erythema assessed by clinicians. On the other hand, two image-sampling methods, based on either selecting specific pixels or small skin areas, performed similarly well. They achieved high intra- and inter-rater reliability with the intraclass correlation coefficient (ICC) and Krippendorff's alpha well above 0.90. These results indicated that smartphone-based dermatoscopy could be a convenient and precise way to evaluate skin erythema severity. However, better outlined, or even automated ISMs, are likely to improve the intra- and inter-rater reliability in severe erythematous cases.

Evaluation of skin erythema severity by dermatoscopy in dogs with atopic dermatitis.

BACKGROUND: To estimate the extent and severity of atopic dermatitis (AD)-related skin lesions, clinical trials enrolling dogs with AD often use categorical scales such as the Canine Atopic Dermatitis Extent and Severity Index, 4th iteration (CADESI-04) and Canine Atopic Dermatitis Lesion Index (CADLI). Despite recent progress in the standardization of these AD-grading scales, the evaluation of the severity of skin lesions (including erythema) remains subjective. OBJECTIVES: To validate an optical set-up with a smartphone and a dermatoscope for the objective estimation of skin erythema severity in atopic dogs. ANIMALS: Forty-three dogs with AD. METHODS AND MATERIALS: An erythema index (EI) was calculated from calibrated skin images and compared to the dermatologist's erythema severity estimate using the erythema grading scale used in the CADESI-04, as well as an ad hoc Visual Analog Scale (VAS) with a continuous palette of red shades. RESULTS: We found a strong correlation based on the Spearman rank correlation coefficient between all erythema valuations: CADESI-04 and VAS: 0.93 [95% CI: (0.85, 0.96)]; CADESI-04 and EI: 0.85 (0.72, 0.92); VAS and EI: 0.82 (0.67, 0.91). There was a good agreement between the objective EI and CADESI-04-based estimates because 71% of samples were classified in the same erythema severity category. When comparing the EI and the VAS, the standard deviation of misestimates was 12% (maximum 100%). CONCLUSIONS AND CLINICAL RELEVANCE: The proposed optical set-up has the potential to make erythema severity estimation objective, thus leading to more reliable AD severity scales for the use in experimental canine AD models or in clinical trials enrolling atopic dogs.

Accuracy of an Affordable Smartphone-Based Teledermoscopy System for Color Measurements in Canine Skin.

Quality smartphone cameras and affordable dermatoscopes have enabled teledermoscopy to become a popular medical and veterinary tool for analyzing skin lesions such as melanoma and erythema. However, smartphones acquire images in an unknown RGB color space, which prevents a standardized colorimetric skin analysis. In this work, we supplemented a typical veterinary teledermoscopy system with a conventional color calibration procedure, and we studied two mid-priced smartphones in evaluating native and erythematous canine skin color. In a laboratory setting with the ColorChecker, the teledermoscopy system reached CIELAB-based color differences ΔE of 1.8-6.6 (CIE76) and 1.1-4.5 (CIE94). Intra- and inter-smartphone variability resulted in the color differences (CIE76) of 0.1, and 2.0-3.9, depending on the selected color range. Preliminary clinical measurements showed that canine skin is less red and yellow (lower a* and b* for ΔE of 10.7) than standard Caucasian human skin. Estimating the severity of skin erythema with an erythema index led to errors between 0.5-3%. After constructing a color calibration model for each smartphone, we expedited clinical measurements without losing colorimetric accuracy by introducing a simple image normalization on a white standard. To conclude, the calibrated teledermoscopy system is fast and accurate enough for various colorimetric applications in veterinary dermatology.

Comparison between rectal and body surface temperature in dogs by the calibrated infrared thermometer.

Because dogs tolerate conventional rectal temperature measurements poorly, a calibrated infrared thermometer was tested for assessing canine body surface temperature. Body surface temperature of 204 dogs was estimated on various sites (digit, snout, axilla, eye, gum, inguinal region, and anal verge). Having rectal temperature as the gold standard, temperature difference, Spearman's correlation coefficient, hyperthermia and hypothermia detection sensitivity and specificity, and stress response score was calculated for each measurement site. Although the canine body surface temperature was considerably lower than the rectal temperature, there was a moderate correlation between both temperatures. Spearman's coefficients were 0.60 (p < 0.001) for the inguinal region with a single operator and 0.50 (p < 0.001) for the gum with multiple operators. Measurement site on the gum additionally guaranteed hyperthermia detection sensitivity and specificity up to 90.0% (95% CI: [66.7 100]) and 78.6% (95% CI: [71.6 85.2]), respectively. Measurements with the infrared thermometer provoked a statistically significant lower stress response (mean stress scores between 1.89 and 2.48/5) compared to the contact rectal measurements (stress score of 3.06/5). To conclude, the correct body surface temperature measurement should include a calibrated thermometer, reliable sampling, and the control of external factors such as ambient temperature influence. The transformation of body surface temperature to the recognized rectal temperature interval allows more straightforward data interpretation. The gum temperature exhibited the best clinical potential since the differences to rectal temperatures were below 1°C, and hyperthermia was detected with the sensitivity of up to 90%.

Photoplethysmography in dogs and cats: a selection of alternative measurement sites for a pet monitor.

OBJECTIVE: Photoplethysmography (PPG) is an increasingly popular health and well-being tool for monitoring heart rate and oxygen saturation. Due to the pigmentation and hairiness of dogs and cats, a pulse oximeter is routinely placed solely on the tongue. As this approach is feasible only for pet monitor use during surgical procedures, we investigate PPG signal quality on several other measurement sites that would be better tolerated by conscious animals. APPROACH: Acquired PPG signals are analyzed by four signal quality indices: mean baseline, signal power, kurtosis, and tolerance score. MAIN RESULTS: In dogs, the metacarpus and tail can be substituted for oral pulse oximeter placement since both measurement sites exhibited high PPG signal kurtosis and were considered well-tolerated. In cats, the digit could be used with some limitations. SIGNIFICANCE: Pet monitors with pulse oximeter probes adjusted to promising measurement sites could enable veterinarians and owners to monitor animals when fully awake.

Detection of canine skin and subcutaneous tumors by visible and near-infrared diffuse reflectance spectroscopy.

Cancer is the main cause of canine morbidity and mortality. The existing evaluation of tumors requires an experienced veterinarian and usually includes invasive procedures (e.g., fine-needle aspiration) that can be unpleasant for the dog and the owner. We investigate visible and near-infrared diffuse reflectance spectroscopy (DRS) as a noninvasive optical technique for evaluation and detection of canine skin and subcutaneous tumors ex vivo and in vivo. The optical properties of tumors and skin were calculated in a spectrally constrained manner, using a lookup table-based inverse model. The obtained optical properties were analyzed and compared among different tumor groups. The calculated parameters of the absorption and reduced scattering coefficients were subsequently used for detection of malignant skin and subcutaneous tumors. The detection sensitivity and specificity of malignant tumors ex vivo were 90.0% and 73.5%, respectively, while corresponding detection sensitivity and specificity of malignant tumors in vivo were 88.4% and 54.6%, respectively. The obtained results show that the DRS is a promising noninvasive optical technique for detection and classification of malignant and benign canine skin and subcutaneous tumors. The method should be further investigated on tumors with common origin.

Properties of contact pressure induced by manually operated fiber-optic probes.

We assess the properties of contact pressure applied by manually operated fiber-optic probes as a function of the operator, probe contact area, and sample stiffness. First, the mechanical properties of human skin sites with different skin structures, thicknesses, and underlying tissues were studied by in vivo indentation tests. According to the obtained results, three different homogeneous silicone skin phantoms were created to encompass the observed range of mechanical properties. The silicon phantoms were subsequently used to characterize the properties of the contact pressure by 10 experienced probe operators employing fiber-optic probes with different contact areas. A custom measurement system was used to collect the time-lapse of diffuse reflectance and applied contact pressure. The measurements were characterized by a set of features describing the transient and steady-state properties of the contact pressure and diffuse reflectance in terms of rise time, optical coupling, average value, and variability. The average applied contact pressure and contact pressure variability were found to significantly depend on the probe operator, probe contact area, and surprisingly also on the sample stiffness. Based on the presented results, we propose a set of practical guidelines for operators of manual probes.

Impact of contact pressure-induced spectral changes on soft-tissue classification in diffuse reflectance spectroscopy: problems and solutions.

Review of the existing studies on the contact pressure-induced changes in the optical properties of biological tissues showed that the reported changes in transmittance, reflectance, absorption, and scattering coefficient are vastly inconsistent. In order to gain more insight into the contact pressure-induced changes observed in biomedical applications involving common probe-spectrometer diffuse reflectance measurement setups and provide a set of practical guidelines minimizing the influence of the changes on the analysis of acquired spectra, we conducted a series of in vivo measurements, where the contact pressure was precisely controlled, and the spectral and contact pressure information were acquired simultaneously. Classification of three measurement sites on a human hand, representing the natural variability in the perfusion and structure of the underlying tissue, was assessed by training and evaluating classifiers at different contact pressure levels and for different probe operators. Based on the results, three practical guidelines have been proposed to avoid classification performance degradation. First, the most suitable pressure level should be identified. Second, the pressure level should be kept in a narrow range during the acquisition of spectra. Third, applications utilizing probes equipped with a calibrated spring can use several classifiers trained at different contact pressure levels to improve classification performance.

Contact pressure-aided spectroscopy.

Contact pressure induced by manually operated fiber optic probes can significantly affect the optical properties of the studied tissue. If the contact pressure and the changes in optical properties are measured properly, then the complementary information can be used to obtain additional insight into the tissue physiology. However, as reliable assessment of the contact pressure in the existing diffuse reflectance setups is difficult, the impact of contact pressure is usually neglected or considered as a source of errors. We introduce a measurement system for controlled application of contact pressure and for the acquisition of diffuse reflectance spectra, which is suitable for in vivo studies and for overcoming the limitations of the existing measurement setups. A spectral-contact-pressure plane is proposed to present the combined information, highlighting the unique tissue response to the applied pressure.

Pressure-induced near infrared spectra response as a valuable source of information for soft tissue classification.

Acquiring near infrared spectra in vivo usually requires a fiber-optic probe to be pressed against the tissue. The applied pressure can significantly affect the optical properties of the underlying tissue, and thereby the acquired spectra. The existing studies consider these effects to be distortions. In contrast, we hypothesize that the pressure-induced spectral response is site- and tissue-specific, providing additional information for the tissue classification. For the purpose of this study, a custom system was designed for dynamic pressure control and rapid acquisition of spectra. The pressure-induced spectral response was studied at three proximate skin sites of the human hand. The diffuse reflectance and scattering were found to decrease with the applied contact pressure. In contrast, the concentrations of chromophores, and consequently the absorption, increased with the applied contact pressure. The pressure-induced changes in the tissue optical properties were found to be site-specific and were modeled as a polynomial function of the applied contact pressure. A quadratic discriminant analysis classification of the tissue spectra acquired at the three proximate skin sites, based on the proposed pressure-induced spectral response model, resulted in a high (90%) average classification sensitivity and specificity, clearly supporting the working hypothesis.