Cutaneous Pythiosis in 2 Dogs, Italy.

We report cutaneous pythiosis in 2 dogs in Italy that had recurrent exposure to the same freshwater habitat. Phylogenetic analysis placed the isolates within Pythium insidiosum complex cluster IV, corresponding to P. periculosum. In Italy, pythiosis should be considered in differential diagnoses by human and veterinary health professionals.

Interdisciplinary approach to solve unusual mortalities in the European common frog (Rana temporaria) in two high-mountain ponds affected by climate change.

The global decline in amphibian populations is a major environmental issue. Chytridiomycosis, Ranaviruses and the red-leg syndrome have been identified in unusual mortality events. However, these infections do not account for all causes of declining amphibian populations. Moreover, several cases of amphibian mortality are difficult to solve without resorting to an interdisciplinary approach. Two cases of unusual mortality in Rana temporaria occurred at two high-mountain ponds (northwest Italy) in April and May 2021. Water and frog samples were analysed to understand the possible causes responsible for the unusual mortalities. Results of the main physicochemical (pH, conductivity, dissolved oxygen, chemical and biochemical oxygen demand) and nutrient (ammonia/ammonium, nitrite, nitrate, total phosphorus) parameters revealed a good condition of the water quality, with the absence of the main cyanotoxins (microcystins/nodularins). However, unseasonably high spring water temperatures were recorded in both ponds (12.73 °C and 14.21 °C for Frog Pond and Selleries Pond, respectively). Frogs (n = 50; snout-vent length: 7.0-9.8 cm; body mass: 85-123 g) collected from Frog Pond mainly presented bumps on the ventral cavity and dermal ulceration associated with the isolation of Carnobacterium maltaromaticum. On the other hand, frogs (n = 5; snout-vent length: 8.0-9.1 cm; body mass: 87-92 g) from Selleries Pond presented petechiae and dermal ulcerations on the rear limbs associated with the isolation of Aeromonas salmonicida and A. sobria. In both mortality events, the interdisciplinary approach revealed an association between frog mortalities and the isolation of bacteria. Isolated bacteria are considered opportunistic pathogens, and the high values of the water temperature has certainly led a stress on the frogs, favouring the spread of bacteria and the death of the frogs. Further studies are needed to assess the pathophysiological effects of the opportunistic bacteria here isolated, clarifying the interactions between emerging pathogens and climate change.

Cases of dermatophytosis caused by Trichophyton benhamiae var. luteum and T. europaeum, newly described dermatophytes within the T. benhamiae complex.

Trichophyton benhamiae var. luteum and T. europaeum - recently described dermatophytes within the T. benhamiae complex - were identified in nine cases of dermatophytosis involving guinea pigs, chinchillas and dogs. The diagnosis was obtained through direct hair/scale examination, culture and sequencing of the internal transcribed spacer region of ribosomal DNA.

Trichophyton benhamiae var. luteum et T. europaeum - dermatophytes récemment décrits au sein du complexe T. benhamiae - ont été identifiés dans neuf cas de dermatophytose de cobayes, de chinchillas et de chiens. Le diagnostic a été obtenu par examen direct des poils/écailles, culture et séquençage de la région ITS de l'ADN ribosomique.

Trichophyton benhamiae var. luteum y T. europaeum, dermatofitos recientemente descritos dentro del complejo T. benhamiae, se identificaron en nueve casos de dermatofitosis que involucraron a cobayas, chinchillas y perros. El diagnóstico se obtuvo a través del examen directo de pelo/escamas, cultivo y secuenciación de la región espaciadora transcrita interna del DNA ribosómico.

Trichophyton benhamiae var. luteum e T. europaeum - dermatófitos recém descritos dentro do complexo T. benhamiae - foram identificados em nove casos de dermatofitoses envolvendo porquinhos da Índia, chichilas e cães. O diagnóstico foi obtido por exame direto de pelos e escamas, cultura e sequenciamento da região espaçadora transcrita interna do DNA ribossomal.

In vitro and in vivo evaluation of a new phytotherapic blend to treat acute externa otitis in dogs.

Canine otitis externa is frequently encountered in veterinary practice, caused by primary factors with bacteria and yeast overgrowth acting as secondary and perpetuating factors. The pharmacological support includes anti-inflammatory, antimicrobials, and antimycotic drugs, but therapeutic failure and antimicrobial resistance are leading to alternative strategies based on phytotherapic products. This study aimed to evaluate an essential oil blend (Otogen® ) to treat otitis externa in dogs. The experimental design was divided in: (a) an in vitro approach, based on the European Normative UNI EN 1275:2006, to assess the efficacy of the product against the most frequently isolated microorganisms during otitis externa. (b) an in vivo part, 12 owned dogs presenting with acute otitis externa were enrolled. A significant growth reduction (>99.9%) of Malassezia pachydermatis and Candida albicans after 15 min of contact and Pseudomonas aeruginosa after 1 h of incubation was recorded. For Staphylococcus pseudintermedius, 50% of growth reduction were appreciated after 15 min. Results obtained in vivo after 7 days of blend administration, noted a significant improvement of all the considered parameters (most important were head shaking, erythema, and scraping). The results obtained may support the usefulness of the tested phytotherapic blend to manage acute otitis externa in dogs.

In Vivo Observation of Trombiculosis with Fluorescence-Advanced Videodermatoscopy.

Trombiculosis is a skin infestation by larvae of mites of the Trombiculidae family. We used fluorescence-advanced videodermatoscopy to diagnose trombiculosis in a woman in Italy with targetoid patches. This method might be useful for identifying atypical manifestations of trombiculosis.

Sarcoptic Mange of Fox Origin in Multiple Farm Animals and Scabies in Humans, Switzerland, 2018.

Fox-derived Sarcoptes scabiei mites caused an outbreak of mange on a farm in Switzerland in 2018. Pruritic skin lesions suggestive of S. scabiei mite infestation developed in 4 humans who had direct contact with affected farm animals but not foxes. Sarcoptic mange is continuously spreading; such outbreaks affecting humans could start occurring more frequently.

Comparison of two inoculation methods for Microsporum canis culture using the toothbrush sampling technique.

BACKGROUND: The fungal culture toothbrush method is a common method for obtaining material for fungal cultures to diagnose dermatophytosis. The optimal technique for inoculation onto the agar surface has not been studied. HYPOTHESIS/OBJECTIVES: To compare two inoculation techniques; the first involved pressing the toothbrush onto the plate surface (Procedure A) and the second involved pressing the toothbrush onto the agar, as well as transferring hairs and scales entrapped in the bristles. (Procedure B). The hypothesis was that transferring hairs onto the plate would increase the likelihood of obtaining positive cultures. ANIMALS: Twenty-six cattery-housed cats were sampled using the toothbrush technique. Two toothbrush samples were obtained from each cat. METHODS AND MATERIALS: The two toothbrush samples from each cat were randomized to Procedure A or B, and the investigator was blinded to inoculation technique. Cultures were performed on a medium specific for dermatophytes. The number of positive plates, and the presence and abundance of colonies of dermatophytes and contaminant moulds were compared between the two techniques. RESULTS: Twenty-one cats were culture-positive for Microsporum canis. Procedure A resulted in a significantly higher number (P < 0.01) of positive plates (20 of 21; 95%) compared with Procedure B (seven of 21; 33%). These results were due mainly to higher plate invasion by contaminant moulds, using Procedure B. CONCLUSIONS AND CLINICAL IMPORTANCE: Based upon the findings of this study, the optimum inoculation technique is to press toothbrush bristles onto agar plates to maximize growth of M. canis and minimize introduction of contaminant inoculation.

Tinea corporis caused by Trichophyton equinum in a rider and review of the literature.

BACKGROUND: Trichophyton equinum is a zoophilic dermatophyte usually causing ringworm in horses. Cases of skin infections in humans are very rare. CASE REPORT : ​We describe a case of tinea corporis caused by T. equinum on a buttock of a rider who often rode bareback with short trousers. CONCLUSION: Although T. equinum was considered in the past as a strict zoophilic dermatophyte, our case report confirms that it can cause dermatophytoses also in humans. A review of epidemiological and clinical features of human infections caused by T. equinum is presented.

Scrotal granulomatous aspergillosis in a dromedary camel (Camelus dromedarius).

BACKGROUND: This report describes a case of primary subcutaneous aspergillosis in a 7-year-old neutered male dromedary camel (Camelus dromedarius). CASE PRESENTATION: The animal developed a large nodular lesion in the right scrotum two years after surgical intervention for neutering. The mass had a firm consistency and was painful at palpation. Histopathology revealed dermal granulomatous inflammation with a necrotic centre, surrounded by plasma cells, macrophages, neutrophils, and sparse fungal hyphae characterised by parallel cell walls, distinct septa, and dichotomous branching. Fungal culture was not performed, but a panel of mono- and polyclonal antibodies specific for different fungal genera identified the hyphae as Aspergillus sp. CONCLUSIONS: The occurrence of subcutaneous lesions is a rare manifestation of aspergillosis in animals, and this appears to be the first case reported in the dromedary camel.

Dermoscopic features in 12 cats with dermatophytosis and in 12 cats with self-induced alopecia due to other causes: an observational descriptive study.

BACKGROUND: Dermoscopy is a noninvasive technique allowing rapid magnified in vivo observation of the skin and structures that lie beneath the skin surface. Various congenital and acquired hair shaft abnormalities may also be evaluated by dermoscopy. Additionally, characteristic features of Microsporum canis-induced tinea capitis and trichotillomania in humans have been reported. OBJECTIVES: To describe the dermoscopic findings observed in cats with patchy alopecia due to M. canis infection and in cats with self-inflicted hair loss. ANIMALS: Twenty-four client-owned cats presented at a veterinary referral practice. METHODS: Dermoscopy was performed with a hand-held nonpolarized light dermoscope at 10-fold magnification. The glass plate of the dermoscope was applied gently to the lesions and no sedation was required. RESULTS: Twelve cats were diagnosed with dermatophytosis and 12 with self-induced alopecia due to other causes. At 10-fold magnification, the most characteristic findings observed in circumscribed lesions of cats with dermatophytosis were opaque, slightly curved, broken hairs of a homogeneous thickness (comma-like structures) and a variable amount of brown-to-yellow greasy scales. In cats with self-induced alopecia, multiple hairs with a normal shaft cleanly broken at different lengths, short tufts of hairs broken at an equal level and hook-like and coiled hairs were observed. CONCLUSIONS AND CLINICAL IMPORTANCE: This observational descriptive study suggests that dermoscopy may represent a helpful noninvasive in vivo technique in the differential diagnosis of patchy alopecia in cats.

A pilot study of the efficacy of wipes containing chlorhexidine 0.3%, climbazole 0.5% and Tris-EDTA to reduce Malassezia pachydermatis populations on canine skin.

BACKGROUND: Wipes containing chlorhexidine and azole derivates have been recommended for veterinary use. No study has been published about their activity against Malassezia pachydermatis. HYPOTHESIS/OBJECTIVES: To evaluate the in vivo and in vitro activity of wipes soaked in a chlorhexidine, climbazole and Tris-EDTA solution against Malassezia pachydermatis. ANIMALS: Five research colony shar-pei dogs. METHODS: Wipes were applied once daily onto the left axilla, left groin and perianal area (protocol A), and twice daily on the right axilla, right groin and umbilical region (protocol B) for 3 days. In vivo activity was evaluated by quantifying Malassezia colonies through contact plates on the selected body areas before and after wipe application. The activity of the solution in which the wipes were soaked was assessed in vitro by contact tests following the European Standard UNI EN 1275 guidelines. RESULTS: Samples collected after wipe application showed a significant and rapid reduction of Malassezia yeast CFU. No significant difference in the Malassezia reduction was found between protocols A and B. In vitro assay showed 100% activity against Malassezia yeasts after a 15 min contact time with the wipe solution. CONCLUSIONS AND CLINICAL IMPORTANCE: Wipes containing chlorhexidine, climbazole and Tris-EDTA substantially reduced the M. pachydermatis population on the skin of dogs. The results, although this was an uncontrolled study performed on a small number of dogs, suggest that these wipes may be useful for topical therapy of Malassezia dermatitis involving the lips, paws, perianal area and skin folds.

In vitro effects of different ozone preparations on microorganisms responsible for endometritis in the mare.

Infectious endometritis is considered one of the major causes of infertility and it can affect up to 60% of barren mares. It is characterized by the presence of one or more microorganisms in the reproductive tract and it is treated with the administration of antibiotics, ecbolic agents and uterine lavages. Ozone, thanks to its antimicrobial properties that are based on its high oxidative potential, could represent an effective alternative treatment for endometritis. The aim of this study was to test in vitro the bactericidal and fungicidal properties of different ozone formulations, either as gas (experiment 1) or dissolved in two liquid matrices (experiment 2), specifically distilled water or oil (Neozone 4000, Cosmoproject, Parma, Italy), onto 6 different species of microorganisms isolated from mares with clinical endometritis, namely Escherichia coli, Staphylococcus aureus, Streptococcus equi subsp. Zooepidemicus, Pseudomonas aeruginosa, Klebsiella pneumoniae and Candida albicans. In the first experiment, 3 clinical antibiotic-resistant strains per each species were exposed to different conditions: to O2O3 gas mixtures (15 and 40 µg/ml for 1, 3 and 5 min), to 100 % O2 or left untreated. The results showed a reduction of the microbial count of over 99,9% for every pathogen, time and concentration of O2O3 gas mixtures tested. Furthermore, gaseous ozone showed both a time-dependant effect (5 vs 3 vs 1 min of exposure) and a concentration-dependant effect (40 vs 15 µg/ml) at 1 and 3 min, while after 5 min no differences were observed. In the second experiment, minimum inhibitory concentration (MIC), and minimum bactericidal/fungicidal concentration (MBC, MFC) of ozonated distilled water and ozonated oil were evaluated. Ozonated oil showed a bactericidal/fungicidal activity against all the strains tested (MIC range 12.5-25 % v/v, MBC/MFC range 12.5-50 % v/v) while ozonated distilled water didn't show an observable antimicrobial effect, discouraging its use as an antimicrobial agent for the treatment of endometritis. The results of this in vitro study indicate that both gaseous ozone and ozonated oil exerted remarkable antimicrobial activities and are promising alternative treatments for infectious endometritis, even when caused by antibiotic-resistant bacteria, and encourage further experiments in an effort to scale down or even prevent the use of antibiotics in equine reproduction.

Sarcoptic mange in Felidae: does Sarcoptes scabiei var. felis exist? A first molecular study.

Domestic and wild felids are considered suitable hosts for the parasitic mite Sarcoptes scabiei, and sarcoptic mange is reported in several felid species in the scientific literature. However, the historic classification of Sarcoptes mites into host-specific varieties does not include S. scabiei var. felis. It is unclear whether sarcoptic mange transmission in felids involves canids, other sympatric species, or exclusively felids. This study aimed to characterize the genetic structure of S. scabiei mites from domestic cats (Felis catus) and Eurasian lynx (Lynx lynx carpathicus), comparing them with Sarcoptes mites from sympatric domestic and wild carnivores. Ten Sarcoptes microsatellite markers were used to genotype 81 mites obtained from skin scrapings of 36 carnivores: 4 domestic cats, one dog (Canis lupus familiaris), 4 Eurasian lynx, 23 red foxes (Vulpes vulpes), and 4 grey wolves (Canis lupus lupus) from either Italy, Switzerland or France. Two genetic clusters of S. scabiei with a geographical distribution pattern were detected: mites from cats originating from Central Italy clustered with those from sympatric wolves. In contrast, all the other mites from Switzerland, France and Northern Italy clustered together. These results strengthen the previously advanced hypothesis that genetic variants of S. scabiei have a predominant geographic-related distribution with cryptic transmission patterns. These patterns may rely on the interactions between different hosts living in the same ecological niche rather than a simple infection among hosts belonging to the same taxon, reinforcing the idea that the S. scabiei historic classification into "var" might have little ongoing relevance.

Title: La gale sarcoptique chez les félidés : Sarcoptes scabiei var. felis existe-t-il ? Première étude moléculaire. Abstract: Les félidés domestiques et sauvages sont considérés comme des hôtes appropriés pour l'acarien parasite Sarcoptes scabiei, et la gale sarcoptique est signalée chez plusieurs espèces de félidés dans la littérature scientifique. Cependant, la classification traditionnelle des acariens du genre Sarcoptes en variétés spécifiques à l'hôte n'inclut pas S. scabiei var. felis. On ne sait pas si la transmission de la gale sarcoptique chez les félidés implique des canidés, d'autres espèces sympatriques ou exclusivement des félidés. Cette étude visait à caractériser la structure génétique des acariens S. scabiei des chats domestiques (Felis catus) et du lynx eurasien (Lynx lynx carpathicus), en les comparant aux Sarcoptes des carnivores domestiques et sauvages sympatriques. Dix marqueurs microsatellites de Sarcoptes ont été utilisés pour génotyper 81 acariens issus de grattages cutanés de 36 carnivores : 4 chats domestiques, un chien (Canis lupus familiaris), 4 lynx eurasiens, 23 renards roux (Vulpes vulpes) et 4 loups gris (Canis lupus lupus) d'Italie, de Suisse ou de France. Deux groupes génétiques de S. scabiei, qui suivent un modèle de distribution géographique, ont été détectés. Les acariens des chats originaires du centre de l'Italie se regroupent avec ceux des loups sympatriques. En revanche, tous les autres acariens de Suisse, de France et d'Italie du Nord sont groupés ensemble. Ces résultats renforcent l'hypothèse précédemment avancée selon laquelle les variants génétiques de S. scabiei ont une distribution géographique prédominante avec des schémas de transmission cryptiques. Ces modèles peuvent reposer sur les interactions entre différents hôtes vivant dans la même niche écologique plutôt que sur une simple transmission parmi des hôtes appartenant au même taxon, renforçant l'idée que la classification historique de S. scabiei en "var" a peu de pertinence.

Human adaptation and diversification in the Microsporum canis complex.

The Microsporum canis complex consists of one zoophilic species, M. canis, and two anthropophilic species, M. audouinii and M. ferrugineum. These species are the most widespread zoonotic pathogens causing dermatophytosis in cats and humans worldwide. To clarify the evolutionary relationship between the three species and explore the potential host shift process, this study used phylogenetic analysis, population structure analysis, multispecies coalescent analyses, determination of MAT idiomorph distribution, sexual crosses, and macromorphology and physicochemical features to address the above questions. The complex of Microsporum canis, M. audouinii and M. ferrugineum comprises 12 genotypes. MAT1-1 was present only in M. canis, while the anthropophilic entities contained MAT1-2. The pseudocleistothecia were yielded by the mating behaviour of M. canis and M. audouinii. Growth rates and lipase, keratinolysis and urea hydrolytic capacities of zoophilic M. canis isolates were all higher than those of anthropophilic strains; DNase activity of M. ferrugineum exceeded that of M. canis. The optimum growth temperature was 28 °C, but 22 °C favoured the development of macroconidia. Molecular data, physicochemical properties and phenotypes suggest the adaptation of zoophilic M. canis to anthropophilic M. ferrugineum, with M. audouinii in an intermediate position.

Microsporum mirabile and its teleomorph Arthroderma mirabile, a new dermatophyte species in the M. cookei clade.

A novel dermatophyte species is described in the Microsporum cookei clade. It differs significantly from known taxa in the two molecular markers analyzed, i.e., ITS and partial ß-tubulin (BT2). Morphologically the species was characterized by smooth- or only slightly rough-walled conidia, but isolates rapidly became pleomorphic with sparse, smooth- and thick-walled macroconidia in addition to microconidia. A teleomorph was found after mating.

Evaluation of the antifungal susceptibility of Malassezia pachydermatis to clotrimazole, miconazole and thiabendazole using a modified CLSI M27-A3 microdilution method.

BACKGROUND: In this study, we evaluated the antifungal susceptibility of Malassezia pachydermatis to clotrimazole (CTZ), miconazole (MCZ), and thiabendazole (TBD), azole derivatives employed in aural formulations labeled for treatment of canine otitis. METHODS: The procedure for in vitro testing was based on the indications of the Clinical and Laboratory Standards Institute (CLSI) M27-A3 microdilution method. A lipid-enriched medium was employed to enhance the yeast growth (Christensen's urea broth, with 0.1% Tween 80 and 0.5% Tween 40 as the lipid sources), while the inoculums size corresponded to approximately 1-5 × 10(5) yeast cells/mL. Microplates were incubated at 37°C and read 48 h after inoculation. Azole MICs inhibiting fungal growth were the lowest drug concentrations that showed an optical density of ≤ 50% of the (drug-free) growth control, as assessed by spectrophotometer (630 nm filter). RESULTS: All isolates were inhibited by the three azoles, with different minimum inhibitory concentration (MIC) values. Most isolates were inhibited by drug concentrations of 2-8 (CTZ), 1-4 (MCZ), or 16-32 (TBD) µg/mL. These results are partially in agreement with the findings of previous studies, in which substantially higher/lower MICs were occasionally reported. This is likely because of the different methodologies employed. Such discrepancies may not apply to clinical situations, where the compounds are applied topically. CONCLUSION AND CLINICAL IMPORTANCE: The concept that clinical failure is linked to increased MICs is debatable, because significantly higher concentrations (in most cases at least 1,000 × the MIC) of the antifungals that were included in our study are routinely used in formulated products.

A case of an apparent infestation by Proisotoma spp. springtails (Collembola: Isotomidae) in a cat.

This case report is presumed to be the first case of infestation of a cat by springtails which are small arthropods closely related to insects. The organisms, found by skin scrapings, were identified as Proisotoma spp. (Collembola: Isotomidae). The cat presented with dermatological lesions (itchy, furfuraceous dermatitis), and we speculate that they were due to this infestation. The pathogenic role of the Collembola was hypothesized because of the large number of organisms, the presence of eggs indicating an active reproduction cycle, the lack of other pathogens (fleas, mites or lice) and the clinical recovery accompanied by the disappearance of Collembola following treatment. The owner seemed to be affected by the infestation, because a few days after having purchased the cat, she developed a pruriginous papular dermatitis on the neck and the arms, which disappeared shortly after treatment of the kitten and a careful washing of all of its toys and other accoutrements.

Re-discovery of Trichophyton bullosum in North Africa as a cause of severe dermatophytosis in donkeys.

This article reports the first verified cases of infection by Trichophyton bullosum in Africa since the description of the fungus, isolated in 1933 from the coat of horses in Tunisia and Mali. We found the fungus in cutaneous samples obtained from donkeys suffering from severe dermatitis with areas of alopecia and scaling in the surroundings of Cairo (Egypt). Fungal elements (arthroconidia and hyphae) were seen at the microscopy of material collected by skin scraping and digested in NaOH. Fungal colonies grown on various culture media were identified through PCR and sequencing of the ITS rDNA region. Since the original report in Africa and the Middle East, only a few cases have been reported thus far in humans in France and two cases in horses in the Czech Republic and Japan. Trichophyton bullosum seems thus an infrequent cause of dermatophytosis. However, the actual prevalence of this pathogen may be underestimated due to the similarity with T. verrucosum, a predominant cause of infection in cattle, occasionally found on horses and donkeys. Indeed, the two fungi can be distinguished only via molecular methods, which are poorly employed in epidemiological studies on equine and bovine dermatophytosis. The present study results add to our knowledge on the ecology of this poorly explored dermatophyte, supporting the concept that equines are the primary hosts of T. bullosum and confirming the presence of this pathogen in Africa. At the same time, these are the first unequivocally documented infections in donkeys due to T. bullosum.

Concurrent Chorioptic Mange and Dermatophytosis in Dairy Goats: A Case Report.

A concurrent chorioptic mange and dermatophytosis outbreak occurred in a goat flock in northwestern Italy. Sanitation of the flock was obtained following pour-on eprinomectin application at a dose of 1 mg/kg; enilconazole was used for environmental disinfection against dermatophyte spores.