Outcome of parasitological examinations in dogs in Germany: a retrospective survey.

Dog faecal samples examined from January 2019 to December 2019 were retrospectively analysed for frequency of endoparasites. The examinations were performed with several different methods: 29,219 samples were examined by flotation method and sodium acetate-acetic acid-formalin concentration (SAFC) technique, 1,330 samples by Baermann-Wetzel migration technique, 12,221 samples using a Giardia coproantigen enzyme-linked-immunosorbent assay (ELISA), 1,180 samples using a Cryptosporidium coproantigen ELISA, 1,671 samples by polymerase chain reaction (PCR) testing for Giardia duodenalis and 447 samples by PCR testing for Cryptosporidium spp.. A total of 7.1% of the samples were positive for parasites in the microscopical examination using the flotation method and SAFC technique. The parasites found included Cystoisospora spp. (2.8%), Giardia duodenalis (2.3%), Ancylostomatidae (1.8%), Toxocara canis (1.6%), Trichuris vulpis (0.7%), Toxascaris leonina (0.5%), Capillaria spp. (0.2%), Angiostrongylus vasorum (0.2%), Crenosoma vulpis (0.1%), Taeniidae (0.1%), Sarcocystis spp. (0.03%), Dipylidium caninum (0.01%), Diphyllobothrium latum (< 0.01%), Spirurida (< 0.01%) and Opisthorchiidae (< 0.01%). Using the Baermann-Wetzel migration technique, Angiostrongylus vasorum was found in 0.75% and Crenosoma vulpis in 0.3% of the samples. ELISAs for Giardia duodenalis and Cryptosporidium spp. revealed 13.9% and 1.0% positive faecal samples, and Giardia duodenalis and Cryptosporidium spp. PCRs 19.4% and 2.0%, respectively. Dogs in the first year of life were more frequently infected with parasites than older animals. In the microscopic examination using flotation method and SAFC technique, the significantly highest detection rates were found in dogs up to six months of age (p < 0.001).

Frequency of potentially pathogenic bacterial and fungal isolates among 28,887 endometrial samples from mares, with an emphasis on multi-drug resistant bacteria in Germany (2018-2022).

Antimicrobial-resistant bacteria pose a serious threat to the wellbeing of animals and humans. In equine reproduction, endometritis caused by facultative microbial pathogens is a condition, which is usually treated with antibiotics. Data from Germany on prevalence of facultative pathogenic microorganisms cultured in samples from the equine uterus and the frequency of multi-drug resistant (MDR) bacteria is lacking. The aim of the study was to provide representative numbers for both. Microbiological culture results (n = 28,887) of endometrial samples submitted to a large veterinary diagnostic laboratory from 2018-2022 were analyzed. An average of 25.9 % of the culture results showed growth of facultative pathogenic bacteria. The dominant isolated bacteria were ß-hemolytic streptococci (79.7 %) followed by Escherichia (E.) coli variatio haemolytica (5.2 %). E. coli were cultured in 4.3 % of the samples and occurred more often than Klebsiella pneumoniae (3.9 %), Candida species (2.9 %), Pseudomonas aeruginosa (2.0 %), and Staphylococcus aureus (1.5 %). Antibiotic susceptibility testing revealed sensitivity of ß-hemolytic streptococci towards penicillins in almost 100 % of the cultured samples (99.5 %). E. coli-isolates were sensitive to gentamicin in 96.2 % of the cases. The frequency of multidrug-resistant extended spectrum beta-lactamase (ESBL)-positive bacteria and methicillin-resistant Staphylococcus aureus (MRSA) was 3.1 % of all positive culture results. The number of ESBL-positive isolates (n = 159) and MRSA was stable from 2018-2022. In conclusion, the situation regarding occurrence of MDR bacteria in Germany is favorable, but should further be monitored.

Challenges in microbiological identification of aerobic bacteria isolated from the skin of reptiles.

INTRODUCTION: Bacterial pathogens are often involved in dermatitis in reptiles. Exact identification of reptile-specific 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. CONCLUSIONS: Most of the aerobic bacterial isolates cultured from reptile skin achieved high scores by MALDI-TOF MS. However, in the majority of category III isolates MALDI-TOF MS results were different from those of the molecular analysis. This strengthens the need to carefully examine low-scored results for plausibility and to be familiar with the occurrence and morphology of relevant reptile-specific bacterial species (e.g. Devriesea agamarum) as well as with the limits of the database used.

German-Wide Analysis of the Prevalence and the Propagation Factors of the Zoonotic Dermatophyte Trichophyton benhamiae.

For about 10 years, a new variant of the pathogen Trichophyton (T.) benhamiae has appeared in Germany, characterized by a previously unobserved culture phenotype with a strong yellow reverse. A few studies suggest that this new variety is now the most common zoophilic dermatophyte in Germany. The guinea pig is the main carrier. Exact prevalence measurements are not yet available. Thus, the aim of our ongoing study was to collect data on the frequency and geographic distribution of the pathogen and its phenotypes (white and yellow) in humans and guinea pigs throughout Germany. Our former studies have already shown that animals from large breeding farms are particularly heavily affected. In contrast to this, 21 small, private breedings were sampled and husbandry conditions recorded. This placed us in a position to identify propagation factors and to give recommendations for containment. For animals from private breedings, we detected T. benhamiae with a prevalence of 55.4%, which is a reduction of nearly 40% compared with animals from large breeding farms. As risk factors, we identified the type of husbandry and the contact to other breedings. Furthermore, certain animal races, like Rex guinea pigs and races with long hair in combination with curls were predestined for colonization with T. benhamiae due to their phenotypic coat characteristics. A prevalence for infections with T. benhamiae of 36.2% has been determined for symptomatic pet guinea pigs suspected of having dermatophytosis and is comparable to the study of Kraemer et al. showing a prevalence of 34.9% in 2009 in Germany. The prevalence in humans is stable with about 2-3% comparing the data of 2010-2013 and 2018 in Thuringia. The new type of T. benhamiae was by far the most frequent cause in all settings.

[Aerobic bacteria from skin lesions in reptiles and their antimicrobial susceptibility]. / Aerobes Keimspektrum und Resistenzlage bei Hautläsionen von Reptilien.

OBJECTIVE: Bacterial skin infections are common in reptiles. Although many such infections are influenced by multifactorial problems, specific treatment of bacterial infections is an important consideration. The objective of this study was to evaluate the range of aerobic bacteria in skin lesions of reptiles and to determine their antimicrobial susceptibility. MATERIAL AND METHODS: Swabs of skin lesions from 219 reptiles were cultured for aerobic bacteria between January 2017 and June 2018. Isolates were identified based on growth on selective agar plates, biochemical parameters, as well as MALDI-TOF MS. Antibiotic susceptibility testing was carried out using the microdilution method. RESULTS: A total of 306 isolates were identified, mostly gram-negative, including Pseudomonas spp. (n = 48), Citrobacter spp. (n = 31, only in chelonians), aerobic spore-forming bacteria (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), Morganella spp. (n = 11) as well as 78 other gram-negative and 12 other gram-positive bacteria. Colonization with 2 (n = 80) or more (n = 16) bacterial isolates was seen in 96 animals. Antibiotic susceptibility testing was carried out with 208 of the 306 isolated bacteria. Many isolates were sensitive (minimal inhibitory concentration [MIC] in µg/ml ≤ breakpoint) to enro- (E) and marbofloxacin (M): 86.4 % MIC ≤ 0.5 (E) and 95.5 % MIC ≤ 1 (M) for Pseudomonas spp., 86.4 % MIC ≤ 0.5 (E) and 90.9 % MIC ≤ 1 (M) for Citrobacter spp., 75.0 % MIC ≤ 0.5 (E) and 100 % MIC ≤ 1 (M) for Aeromonas spp. Trimethoprim/sulfamethoxazol proved to be effective against most of the Citrobacter spp. (90.9 % MIC ≤ 2/38) and Aeromonas spp. (75.0 % MIC ≤ 2/38). Amikacin was effective against nearly all Pseudomonas spp. (97.7 % MIC ≤ 16), Citrobacter spp. (95.5 % MIC ≤ 16) and Aeromonas spp. (93.8 % MIC ≤ 16). CONCLUSION AND CLINICAL RELEVANCE: The majority of isolates were gram-negative; the clinical relevance of individual isolates must, however, be evaluated on a case by case basis. Many of the isolated bacteria were sensitive to fluoroquinolones as well as aminoglycosides. Susceptibility testing is recommended since use of these antibiotics should be limited and for every tested group of antibiotics resistant isolates were found.

Antifungal in vitro Activity of Essential Oils against Clinical Isolates of Malassezia pachydermatis from Canine Ears: A Report from a Practice Laboratory.

BACKGROUND: The yeast Malassezia pachydermatis is a commensal but also the most isolated pathogenic yeast in canine otitis externa. Essential oils (EOs) exhibit antimicrobial activity and thus could be an alternative for treatment of otitis. OBJECTIVE: The aim of this study was to analyse the in vitro efficacy of EOs against M. pachydermatis isolates from canine ears. MATERIALS AND METHODS: Twenty-two EOs were tested for their in vitro activity against 15 M. pachydermatis isolates by agar disc diffusion. In addition, diluted EOs were tested and volatile antifungal activity was analysed by vapour assay. RESULTS: Most EOs showed in vitro efficacy against M. pachydermatis. A different susceptibility pattern of tested isolates was found. Overall, the most potential EO was lemon grass. Also, cinnamon leaf, clove, manuka, Indian melissa, oregano, palmarosa, and winter savory oil exhibited a strong antifungal activity. Volatile activity was proven by the inhibiting effect in vapour assay. CONCLUSION: EOs show in vitro activity against M. pachydermatis. This study gives scientific background for the use of EOs in veterinary medicine. Diverse susceptibility patterns of different M. pachydermatis strains emphasise the helpfulness of an aromatogram as one lead for the best choice of an EO for therapy.

DETECTION OF OPHIDIOMYCES OPHIODIICOLA IN TWO CAPTIVE BOCOURT WATER SNAKES ( SUBSESSOR BOCOURTI) AND ONE CAPTIVE PUEBLAN MILK SNAKE ( LAMPROPELTIS TRIANGULUM CAMPBELLI).

Two captive Bocourt water snakes ( Subsessor bocourti) presented with chronic white skin lesions on their heads; Ophidiomyces ophiodiicola was identified by culture and polymerase chain reaction (PCR) in skin scrapings from both snakes. Histopathology performed in one Bocourt water snake revealed fungal hyphae in epidermal structures of lesions. One Pueblan milk snake ( Lampropeltis triangulum campbelli) from the same zoologic institution presented with yellow crusts and white blisters on its body, from which O. ophiodiicola was identified by culture and PCR. Two of the three snakes apparently recovered from lesions after multiple natural sheds, whereas the third snake died. This is the first report of O. ophiodiicola infection in Bocourt water snakes and in a Pueblan milk snake, as well as the first report of O. ophiodiicola in France.

[Occurrence of Ehrlichia canis in dogs living in Germany and comparison of direct and indirect diagnostic methods]. / Vorkommen von Ehrlichia canis bei in Deutschland lebenden Hunden sowie Vergleich direkter und indirekter Diagnostikmethoden.

OBJECTIVE: To study the occurrence of Ehrlichia (E.) canis in dogs living in Germany and evaluate the possibilities and limits of direct and indirect diagnostic methods. MATERIAL AND METHODS: The first part of the study was a retrospective analysis of routine samples, which had been examined for E. canis antibodies using an indirect immunofluorescence antibody test (IFAT). The examination was part of a laboratory profile for the detection of travel-related diseases (travel disease profile) or was performed on an individual basis. In the second part, samples which were examined within a travel disease profile, including E. canis antibodies, were further evaluated for E. canis DNA by a polymerase chain reaction (PCR). Medical histories were obtained of E. canis-positive dogs (animals which were positive in IFAT and/or PCR) and for a part of the negative animals. RESULTS: For 2015, 11.8% of 12220 samples had antibodies for E. canis. Dogs, which were examined with a travel disease profile, displayed antibodies in 5.6% cases of 1172 animals (investigation period February to March 2016). Of the E. canis-positive dogs (n = 67), 91% were positive with only the IFAT, 7.5% with the IFAT and PCR and 1.5% were only positive by PCR. Anamnesis showed that particularly imported dogs without symptoms were controlled for travel diseases during their first year in Germany. CONCLUSION AND CLINICAL RELEVANCE: Although endemic E. canis infections play a minor role in Germany, infections with this pathogen are still of importance in this country, particularly because of the import of dogs. Therefore, a medical history helps in early ehrlichiosis diagnosis and to start an adequate treatment. Pathogen detection in imported E. canis-seropositive dogs, which often did not display clinical symptoms in this study, was frequently negative in blood samples by PCR. The diagnostic method should be chosen depending on the disease phase and the underlying question (symptoms or preventive screening).

MICs of 32 antimicrobial agents for Rhodococcus equi isolates of animal origin.

OBJECTIVES: The aim of this study was to determine the MICs of 32 antimicrobial agents for 200 isolates of Rhodococcus equi of animal origin by applying a recently described broth microdilution protocol, and to investigate isolates with distinctly elevated rifampicin MICs for the genetic basis of rifampicin resistance. METHODS: The study included 200 R. equi isolates, including 160 isolates from horses and 40 isolates from other animal sources, from the USA and Europe. MIC testing of 32 antimicrobial agents or combinations thereof followed a recently published protocol. A novel PCR protocol for the joint amplification of the three rpoB regions in which rifampicin resistance-mediating mutations have been reported was applied to isolates with elevated rifampicin MICs. The amplicons were sequenced and screened for mutations. RESULTS: Susceptibility testing revealed a rather uniform distribution of MICs for most of the antimicrobial agents tested. The lowest MICs were seen for clarithromycin, rifampicin and imipenem. Six isolates (3%) exhibited distinctly higher MICs of rifampicin than the remaining 194 isolates. In five of these six isolates, single bp exchanges, which resulted in the amino acid exchanges Gln513Leu, Asp516Val, His526Asp or Ser531Leu, were detected in the rifampicin resistance-determining region 1 of the rpoB gene, with Gln513Leu representing a novel substitution for R. equi. CONCLUSIONS: This study shows the MIC distribution of 32 antimicrobial agents for a large collection of R. equi isolates of animal origin from two continents. Isolates that exhibited distinctly elevated MICs of rifampicin were only rarely detected.