Introduction Bacterial pathogens are often involved in dermatitis in reptiles. Exact identification of reptilespecific but otherwise uncommon bacterial species may be challenging. However, identification is crucial to evaluate the importance of the detected bacterial species. Objective The aim of this study was to assess the number of aerobic bacterial isolates cultured from skin-derived samples of reptiles which were not reliably identified by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS), and to determine their identity. Material and methods Routine bacterial diagnostics were performed on 235 skin samples, and 417 bacterial isolates were analysed by MALDI-TOF MS. The isolates were grouped into categories based on their first score: category I (� 2.00), category II (� 1.70 and < 2.00), and category III (< 1.70). Isolates from category III were further investigated by 16S rRNA gene sequencing and the following criteria were applied: query cover 100%, e-value rounded to 0.0 and sequence identity (%) > 98.00% for genus identification, and > 99.00% for species identification. Results The majority of bacterial isolates were in category I (85.1%) or category II (8.4%). In category III (6.5%) results achieved at first by MALDI-TOF MS corresponded to the results of the molecular analysis in 8.0% of isolates at the species level and in 24.0% at the genus level. Bacterial isolates classified as category III were heterogenic in genus (e.g. Chryseobacterium, Devriesea, Pseudomonas, Staphylococcus, Uruburuella), and some have only been described in reptiles so far.
Copy number variations (CNVs) of the KITLG gene seem to be involved in the oncogenesis of digital squamous cell carcinoma (dSCC). The aims of this study were (1) to investigate KITLG CNV in giant (GS), standard (SS), and miniature (MS) schnauzers and (2) to compare KITLG CNV between black GS with and without dSCC. Blood samples from black GS (22 with and 17 without dSCC), black SS (18 with and 4 without dSSC; 5 unknown), and 50 MS (unknown dSSC status and coat colour) were analysed by digital droplet PCR. The results are that (1) most dogs had a copy number (CN) value > 4 (range 2.5–7.6) with no significant differences between GS, SS, and MS, and (2) the CN value in black GS with dSCC was significantly higher than in those without dSCC (p = 0.02). CN values > 5.8 indicate a significantly increased risk for dSCC, while CN values < 4.7 suggest a reduced risk for dSCC (grey area: 4.7–5.8). Diagnostic testing for KITLG CNV may sensitise owners to the individual risk of their black GS for dSCC. Further studies should investigate the relevance of KITLG CNV in SS and the protective effects in MS, who rarely suffer from dSCC.
Zusammenfassung Gegenstand und Ziel Bakterielle Hautinfektionen kommen bei Reptilien häufig vor. Obwohl viele dieser Infektionen durch multifaktorielle Probleme verursacht werden, ist eine spezifische Behandlung nötig. Ziel der Studie war, das Keimspektrum und die Resistenzlage von aeroben Bakterien in Hautläsionen von Reptilien zu untersuchen. Material und Methoden Tupferproben dermaler Läsionen von 219 Reptilien wurden bakteriologisch untersucht (01/2017–06/2018). Die Identifizierung der Bakterien erfolgte anhand von Selektivnährböden, biochemischen Parametern sowie MALDI-TOF-MS, die Erstellung der Antibiogramme mittels Mikrodilutionsmethode. Ergebnisse Bei den insgesamt identifizierten 306 Keimisolaten handelte es sich überwiegend um gramnegative Spezies. Pseudomonas spp. (n = 48), Citrobacter spp. (n = 31, nur bei Schildkröten), aerobe Sporenbildner (n = 30), Aeromonas spp. (n = 20), Acinetobacter spp. (n = 20), Proteus spp. (n = 15), Staphylococcus spp. (n = 15), Klebsiella spp. (n = 13), Enterococcus spp. (n = 13) sowie Morganella spp. (n = 11) machten den Hauptteil aus, daneben konnten weitere gramnegative (n = 78) und grampositive (n = 12) Bakterienspezies identifiziert werden. Mischkulturen mit 2 (n = 80) oder mehr (n = 16) Keimen traten bei 96 Tieren auf. Von 208 der 306 Isolate wurden Antibiogramme erstellt. Gegenüber Enro- (E) und Marbofloxacin (M) waren viele Isolate sensibel (minimale Hemmkonzentration [MHK] in µg/ml ≤ Grenzwert), beispielsweise Pseudomonas spp. (E: 86,4 % MHK ≤ 0,5; M: 95,5 % MHK ≤ 1), Citrobacter spp. (E: 86,4 % MHK ≤ 0,5; M: 90,9 % MHK ≤ 1) und Aeromonas spp. (E: 75,0 % MHK ≤ 0,5; M: 100 % MHK ≤ 1). Trimethoprim/Sulfamethoxazol erwies sich als wirksam gegen die meisten Citrobacter- (90,9 % MHK ≤ 2/38) und Aeromonas- (75,0 % MHK ≤ 2/38) Isolate. Amikacin war wirksam gegen fast alle Pseudomonas spp. (97,7 % MHK ≤ 16), Citrobacter spp. (95,5 % MHK ≤ 16) sowie Aeromonas spp. (93,8 % MHK ≤ 16). Schlussfolgerung und klinische Relevanz Das Keimspektrum von Reptilienhautläsionen umfasst vor allem gramnegative Bakterien, deren klinische Relevanz für jeden Einzelfall abzuwägen ist. Viele Isolate dieser Studie waren sensibel für Fluorchinolone sowie Aminoglykoside. Da der Einsatz dieser Antibiotika zurückhaltend erfolgen sollte und gegenüber jeder getesteten Antibiotikagruppe auch resistente Isolate identifiziert wurden, wird eine Antibiogrammerstellung empfohlen.
Most canine intestinal tumours are B-cell or T-cell lymphomas or carcinomas. They have to be distinguished from cases of enteritis. Non-invasive biomarkers such as miRNAs would be a step towards faster diagnosis. The aim of this study was to investigate shifts in miRNA expression in tissue samples collected from cases of enteritis, carcinoma and lymphoma of the small and large intestine to better understand the potential of miRNA as biomarkers for tumour diagnosis and classification. We selected two oncogenic miRNAs (miR-18b and 20b), two tumour suppressive miRNAs (miR-192 and 194) and two potential biomarkers for neoplasms (miR-126 and 214). They were isolated from FFPE material, quantified by ddPCR, normalised with RNU6B and compared with normal tissue values. Our results confirmed that ddPCR is a suitable method for quantifying miRNA from FFPE material. Expression of miR-18b and miR-192 was higher in carcinomas of the small intestine than in those of the large intestine. Specific miRNA patterns were observed in cases of enteritis, B-cell and T-cell lymphoma and carcinoma. However, oncogenic miR-18b and 20b were not elevated in any group and miR-126 and 214 were down-regulated in T-cell and B-cell lymphoma, as well as in carcinomas and lymphoplasmacytic enteritis of the small intestine.
(1) Background: Devriesea (D.) agamarum is a potential cause of dermatitis and cheilitis in lizards. The aim of this study was to establish a real-time PCR assay for the detection of D. agamarum. (2) Methods: Primers and probe were selected targeting the 16S rRNA gene, using sequences of 16S rRNA genes of D. agamarum as well as of other bacterial species derived from GenBank. The PCR assay was tested with 14 positive controls of different D. agamarum cultures as well as with 34 negative controls of various non-D. agamarum bacterial cultures. Additionally, samples of 38 lizards, mostly Uromastyx spp. and Pogona spp., submitted to a commercial veterinary laboratory were tested for the presence of D. agamarum using the established protocol. (3) Results: Concentrations of as low as 2 × 104 colonies per mL were detectable using dilutions of bacterial cell culture (corresponding to approximately 200 CFU per PCR). The assay resulted in an intraassay percent of coefficient of variation (CV) of 1.31% and an interassay CV of 1.80%. (4) Conclusions: The presented assay is able to detect D. agamarum in clinical samples, decreasing laboratory turn-around time in comparison to conventional culture-based detection methods.
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