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.

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.

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.

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.

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.

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.

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.

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.

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.

Dermanyssus gallinae in non-avian hosts: A case report in a dog and review of the literature.

Non-avian attacks of the worldwide distributed mite Dermanyssus gallinae are occasionally reported. However, it is widely accepted that their occurrence is underestimated. The present study aims to describe the first Italian case of dermanyssosis in a dog, to molecularly characterize the mites collected from the patient and the animal enclosure, where poultry and dog were confined, and to review the current literature on the non-avian attacks by D. gallinae. The dog was successfully treated with an oral sarolaner-based product, followed by a spot-on formulation of imidacloprid and moxidectin. The infestation source was likely attributable to poultry and confirmed by molecular identification of D. gallinae sensu strictu. Ten articles on non-avian D. gallinae attacks in domestic animals and wildlife were retrieved, pointing out the need for more awareness amongst practitioners. The therapeutic effect of available antiparasitic drugs, currently used off-label, should also be better explored in non-avian hosts.

Diagnostic and therapeutic management of Cryptococcosis in a kitten with practical considerations to veterinary pediatric therapeutic approach.

A 3-months-old male domestic kitten was referred for repeated seizures. Analysis revealed Cryptococcus neoformans. Levetiracetam and fluconazole were administered without significant clinical improvements and without negativization. Hypothesizing resistance, therapy was switched to amphotericin B. Seizure disappeared. Haematological controls highlighted transitory increasing of CK, BUN, ALP and cholesterol. PCR repeated two weeks after the treatment was negative for Cryptococcus neoformans. Nowadays the cat is 5 years old, and no seizures occurred since the age of 5 months.

Does the Introduction of Alien Species Represent a Sanitary Threat for Native Species? The Case of the Eastern Cottontail Sylvilagus floridanus in Italy.

Introduction of alien species is a well-known threat to biodiversity. Where newly introduced, alien species may pose a risk for the local ecological community by competing for resources or by introducing pathogens. Sylvilagus floridanus is an American lagomorph introduced into Europe in the second half of 20th century, for hunting. This study evaluated the structure and epidemiological characteristics of the gastrointestinal parasite community in an introduced population of S. floridanus in the Province of Alessandria (Piedmont Region-Italy). Three alien parasites were reported out of 271 animals: Obeliscoides cuniculi in the stomach, Trichostrongylus calcaratus in the small intestine, and Passarulus nonnanulatus in the large intestine. All these nematodes are commonly reported in S. floridanus in its natural range, but they represent alien species in Europe. The report of these alien parasites is an example of the unexpected consequences caused by the introduction of non-native vertebrates. The documented introduction of new pathogens may alter the parasite community of the native lagomorphs, with possible long-term effects on local ecological dynamics.

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.

Antifungal Resistance Regarding Malassezia pachydermatis: Where Are We Now?

Malassezia pachydermatis is a yeast inhabiting the skin and ear canals in healthy dogs. In the presence of various predisposing conditions it can cause otitis and dermatitis, which are treated with multiple antifungal agents, mainly azole derivatives. This manuscript aims to review the available evidence regarding the occurrence of resistance phenomena in this organism. Various findings support the capacity of M. pachydermatis for developing resistance. These include some reports of treatment failure in dogs, the reduced antifungal activity found against yeast isolates sampled from dogs with exposure to antifungal drugs and strains exposed to antifungal agents in vitro, and the description of resistance mechanisms. At the same time, the data reviewed may suggest that the development of resistance is a rare eventuality in canine practice. For example, only three publications describe confirmed cases of treatment failure due to antifungal resistance, and most claims of resistance made by past studies are based on interpretive breakpoints that lack sound support from the clinical perspective. However, it is possible that resistant cases are underreported in literature, perhaps due to the difficulty of obtaining a laboratory confirmation given that a standard procedure for susceptibility testing of M. pachydermatis is still unavailable. These considerations highlight the need for maintaining surveillance for the possible emergence of clinically relevant resistance, hopefully through a shared strategy put in place by the scientific community.

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.

Azole resistance of Malassezia pachydermatis causing treatment failure in a dog.

A 5-year-old neutered female toy Poodle chronically treated with systemic and topical azoles to control recurrent Malassezia dermatitis/otitis was presented because of the loss of treatment efficacy. Minimum Inhibitory Concentrations (MICs) obtained in vitro for various azoles (especially itraconazole and ketoconazole) against Malassezia strains isolated from the dog were increased by several-fold compared with MICs obtained for control isolates. These results reinforced the assumption based on clinical observation, i.e. the development of azole resistance.

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.

Cladosporium cladosporioides-complex infection in a mixed-breed dog.

A 3-year-old female mixed-breed dog was referred with a 2-day history of serious dyspnea, coughing, lethargy, anorexia, and a low-grade right anterior lameness. At presentation, the dog had an increased respiratory rate, dull heart and lung sounds, and cyanotic mucous membranes. It was hyperthermic and slightly dehydrated. Laboratory findings showed mild neutrophilia with a left shift, while serum biochemistry variables were in the normal range. However, urinalysis revealed mild proteinuria and rare erythrocytes and leukocytes on sediment examination. Thoracic radiographs showed a diffuse mixed interstitial and alveolar pattern with an air bronchogram, while appendicular radiographs showed a right humeral interrupted brush-like periosteal reaction. Thoracic ultrasonography revealed mediastinal lymph node enlargement. Cytology from a fine-needle aspirate of mediastinal lymph nodes revealed a pyogranulomatous lymphadenitis with numerous fungal hyphae. Culture on Sabouraud dextrose agar isolated dark fungal colonies with microscopic features consistent with Cladosporium spp. Sequencing of the internal transcribed spacer region identified the fungus as a species of the Cladosporium cladosporioides-complex.

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.

Infection by Microsporum canis in Paediatric Patients: A Veterinary Perspective.

Microsporum canis is a dermatophyte fungus of which cats and dogs are recognized as the natural hosts. M. canis is also easily transmitted to humans, causing lesions to the glabrous skin (tinea corporis) and to the head (tinea capitis). The present study describes some cases of infection with M. canis in children from a veterinary perspective, highlighting some important features of this clinical entity (e.g., the necessity to identify the animal source of infection with appropriate diagnostic tests; the fact that infected cats may present with no or atypical dermatological signs; and the importance of the environment as a fungal reserve).