ABSTRACT: Molds and yeasts are significant components in dairy products and reports have documented the presence of it milk and cheese. This study identified the molds and yeasts in raw milk and cheese in a Metropolitan Area of Porto Alegre, South Brazil. The study included a total of 143 samples (113 of raw milk and 30 of creamed cheese). All samples were obtained between 2020 and 2024. Milk samples came from producers in the Metropolitan Region of South Brazil and cheese samples were directly collected in the commercial wrappings provided by the producers. Fungal isolation was performed using serial dilutions plated on Sabouraud Dextrose Agar and Malt Extract Agar media (25 ºC - 37 ºC for 5 to 7 days). The identification was made by using conventional and molecular methods. Yeasts and molds were isolated from 88/62% of the samples (65 raw milk and 23 cheese samples), being Candida and Aspergillus the most isolated genera. Forty-four yeasts were identified in the genera Candida, Cryptococcus, Rhodotorula, and Trichosporon. Within the Candida genus, the species C. parapsilosis, C. tropicalis and C. albicans were identified. Thirty-one molds from the genera Aspergillus, Penicillium, Geotrichum, Fusarium, and Rhizopus were found. The results showed that the studied products contain different types of yeasts and filamentous fungi. The isolation of these fungal genera demonstrated their direct relevance to dairy product quality and safety, as several species are capable of causing chemical and organoleptic deterioration, reducing shelf life, and posing potential risks due to their ability to produce mycotoxins, emphasizing the importance of fungal monitoring throughout the dairy production chain.

Background: Fungal diseases in bovines might cause considerable economic losses for farmers and the livestock industry due to decreased productivity caused by various factors. The most important causes described in the literature are: direct infections, secondary complications like reduced growth productivity and reproduction, weight loss, decreased milk production, production of mycotoxins, treatment costs, and even mortality. The purpose of this manuscript is to describe a retrospective data about fungal isolated from bovines at the Sector of Veterinary Mycology, School of Veterinary Medicine of the Federal University of Rio Grande do Sul (UFRGS) during a 38-year period of routine diagnosis (1986-2023). Materials, Methods & Results: Two-hundred and thirty-two (n = 232) mycological cultures were made from bovine samples from specimens of various systems [integumentary (n = 84); 27 respiratory (n = 27); reproductive (n = 41) and digestive (n = 36)] and 44 samples of quarter milk from subclinical mastitis. Culture media used were as the following: Sabouraud dextrose agar (SDA) with chloramphenicol; SDA with chloramphenicol & cyclohexamide; and yeast extract-malt agar with chloramphenicol. Trichophyton verrucosum was the most dermatophyte zoophilic diagnosticated from the hair samples. From milk samples the yeast Candida spp. was the most prevalent pathogen isolated. Samples from cattle with respiratory disorders (n = 27) showed the presence of fungi (18 = 67%) with a great majority being hyalohyphomycetes (15 = 55%). In relationship to positive cultures (49 = 64%) from abortion (77 samples), hyalohyphomycetes (n = 37) were the most isolated fungi, comprising 75.5% of the total isolates, with the genus Aspergillus spp. (n = 19; 10 A. fumigatus) being the most prevalent within this group; yeasts (7 = 14%) were the 2nd type of fungi cultured from abortion specimens. Also, some dairy and beef cattle feed samples were analyzed for fungal contamination showing various fungal species: rice straw (Fusarium sp., Mucor sp. and Aspergillus flavus); sorghum silage (Candida albicans); alfalfa (A. flavus, A. niger and Rhizopus sp.); barley (A. clavatus) and feed (A. clavatus). Discussion: The results of fungi isolated from several bovine organs over a period of 38 years are intended to emphasize the need to routinely investigate a possible fungal etiology in various clinical conditions, at least as a differential diagnosis. Mycoses affecting ruminants are usually diagnosed after the animal's death and occur after fungal propagules invade via the airborne, digestive, or intramuscular routes for subsequent hematogenous or lymphatic dissemination to various organs. Due to the fact that the fungi isolated in the present study are ubiquitous, complementary tests, especially histopathological analysis, are often necessary to ascertain with certainty that it is a mycotic condition. Knowledge of the diseases affecting cattle in Brazil, whether caused by bacteria, viruses, fungi, or other etiologies, is very important for decision-making aimed at reducing their occurrence and minimizing the resulting economic losses. The world literature cites parasitic and bacterial diseases, as well as management factors, as the most common causes of losses due to carcass condemnation. Fungi are usually more associated with meat alteration and contamination during refrigerated storage. The fungi, whether filamentous or yeasts, described in the present manuscript are recognized worldwide as being associated with the occurrence of various clinical conditions in several animal species, as well as in humans.

Background: Molds are filamentous saprophytic fungi with worldwide distribution, which are found in various environmental substrates. Among them, the Aspergillus genus stands out as one of the main causes of lung disease. Commercial poultry such as chickens and turkeys, as well as captive and free-ranging wild birds species are most frequently affected. The infection occurs by the inhalation of conidia, which are dispersed in the air by wind. The most frequent acute clinical manifestation is usually observed in young animals, often with episodes of outbreaks in poultry farms, whereas chronic disease is more frequently observed in adult birds. In slaughterhouses, carcass inspection is very important for the detection of certain diseases and for monitoring their occurrence. The objective of this study was to assess the occurrence of aerosacculitis caused by molds in commercial broilers slaughtered in one establishment under Federal Inspection Service (SIF) through mycological, histopathological and molecular diagnosis.Materials, Methods & Results: The samples (n = 76) came from 9 flocks localized in the Vale do Taquari - State of Rio Grande do Sul, RS, Brazil. Lungs were collected from broilers with airsacculitis, directly from the slaughter line and kept under refrigeration (4 oC). The evaluation and condemnation of carcasses were performed by the sanitary inspection department of the slaughterhouse. The molds were identified as the causal agent through fungal culture, histopathology and PCR analysis. Sixteen (21.0%) lung samples were positive in mycological culture, being Aspergillus sections Fumigati and Flavi the most frequent, and 2 (2.7%) samples showed fungal elements (hyaline, branching septate hyphae). The respective isolates were identified as Aspergillus fumigatus sensu stricto following the amplification of β-tub gene fragments and gel electrophoresis. Among the remaining 58 samples: no fungal growth in 61.9% (47/76) cultures and in 14.4% (11/76) were isolated colonies of hyaline septate fungi and of zygomycetes which were considered as common environmental fungi belonged to genera: Aspergillus, Penicillium, Fusarium, Scopulariopsis, Mucor and Rhizopus.Discussion: Poultry become infected through aspiration in large numbers of conidia of molds. According to the literature,A. fumigatus sensu stricto, which belongs to the section Fumigati, is the most ubiquitous in the environment and frequently associated with lung diseases in humans and animals. Identification of fungi to species level is important in order to determine the etiology of the disease and also to detect potential novel agents. Our results corroborate previous studies that showed A. fumigatus to be the prevalent etiological agent identified in commercial poultry. Although in smaller quantities, isolates of the section Flavi were also recovered from the lungs of healthy chickens. In the present study, only A. fumigatus was cultivated from airsacculitis cases. In poultry, many environmental factors may be present and influence the development of aspergillosis, including excessive ammonia and humidity, inadequate temperature and degraded litter. Our results are in agreement with other research where Aspergillus was detected in the lungs of chicken condemned by the inspection service. The eco-epidemiology of Aspergillus sections Fumigati and Flavi needs to be reevaluated, especially considering the transmission from environment to animals and humans. Further studies should be conducted in order to correlate clinical aspects with molecular diversity and antifungal profile among Aspergillus, as well as, investigate mold diversity isolated from commercial poultry in locations where the poultry industry is economically significant. Keywords: moulds, Aspergillus spp., Aspergillus section Fumigati, Aspergillus section Flavi, poultry, airsacculitis.

Background: Sporotrichosis is a subcutaneous mycosis of humans and animals. In recent years, several cities in Brazil have recorded an increase in cases due to zoonotic transmission through cats. The classic clinical presentation is transmitted through plant debris and the 2nd way (zoonotic form) is caused by animal transmission. The alternative zoonotic route has gained attention due to the rapid and significant spread of cat-transmitted sporotrichosis in South America. The aim of this work was to identify the Sporothrix species responsible of feline sporotrichosis based on reference standard and molecular methods. Cases: All isolates were recovered from skin of 27 cats with sporotrichosis between 2020 and 2024. Fifteen cats sampled were from state of Santa Catarina (microregion of Florian & oacute;polis) and, 12 from state of Rio Grande do Sul (11 from microregion of Porto Alegre / 1 from Serafina Corr & ecirc;a city), Brazil. Clinical diagnosis of sporotrichosis was made through examination (inspection and palpation of skin and/or mucous lesions, palpation of superficial lymph nodes and cardiopulmonary auscultation) and cytopathology. The cytopathological examination was performed either by imprint in ulcerated lesions/swabs. Yeast-like structures were observed in cytopathological exam. Fungal growth in mycological culture (Sabouraud Cycloheximide-Chloramphenicol Agar, incubated at 25 degrees C and 37 degrees C for 10 to 30 days) was monitored by forming dark black colonies. Physiological tests (assimilation of sucrose and raffinose), microscopy of hyphae/conidia and macroscopic colony characteristics (colors and texture) were conducted for the identification of the species. All isolates included in the study exhibited similar macroscopic morphology in their filamentous form. Microscopic observation of the isolates showed presence of septate hyphae showing at the end of conidiophores conidia arranged in a "daisy-like" pattern (sympodial formation). Assimilation tests using sucrose and raffinose were negative. Sporothrix brasiliensis was the only agent involved in feline sporotrichosis. The identity of isolates was confirmed using specific primers for the CAL gene and sequencing analyses. Discussion: According to the literature, our result corroborates reports from other authors who describe cases of sporotrichosis caused by Sporothrix brasiliensis in several states in Brazil. Although S. brasiliensis is predominant, the identification of fungal species is essential to determine the etiology of sporotrichosis in different areas of the country, and to indicate a possible change in the current scenario of the disease. The transboundary expansion of the species occurs between states and more cases of cat-cat/cat-dog and cat-human transmission may occur; situation described in this work. In the state of Rio Grande do Sul (RS), S. brasiliensis was the only isolated species. Sporothrix brasiliensis is considered the most virulent species within pathogenic clade and represents an important not only in Brazil, but also in other South American countries; among them are the Paraguay, Argentina and Chile with a description of feline sporotrichosis. In Panama (Central America) the 1st case of zoonotic transmission was reported, however the species of Sporothrix genus was not discriminated through molecular diagnosis. In conclusion, the real situation of sporotrichosis in Brazil can be assessed through the clinical diagnosis of sporotrichosis in felines, as well as in humans that have had zoonotic transmission. In addition, laboratory diagnosis, with the respective determination of the species involved. This way the disease will become more visible and effective control measures can be taken within the scope of public health.

Background: Mycoses in Chelonians have different features; they can be affected by fungi causing shell, tegumental, and/ or systemic disease, which can be considered as conventional agents. Geotrichum species are colonies similar to yeast, producing arthroconidia without blastoconidia. The diseases of the shell are one important disorder that might affect the survival of chelonians. They are more often found in aquatic species kept in captivity and are usually caused by poor environmental conditions. Also, bites and accidents that hurt the shell are factors that allow pathogenic organisms, such as bacteria, fungi, or even algae, to colonize and penetrate the shell. This can lead to conditions called septicemic cutaneous ulcerative disease, which causes deep ulcerations This report describes a case of cutaneous hyalohyphomycosis caused by Geotrichum candidum in a turtle. Case: A 3-year-old female Red-Eared Slider Turtle weighing 0.3 kg attacked by a dog 2 weeks earlier and presenting many deeps wounds with necrotic areas in the carapace as the one relevant clinical feature, was assisted at the Service of Dermatology, Hospital de Cl & iacute;nicas Veterin & aacute;rias, UFRGS, Porto Alegre, Brazil. Samples were obtained from the shell scrapings and the wounded necrotic areas of shell tissues were removed for routine microbiological and histological diagnosis. Direct microscopic examination with mount in 10% KOH revealed the presence septate hyaline hyphae. Bacteriological cultures done on Blood agar and yielded few colonies of Citrobacter freundii. For mycological diagnosis, the material was inoculated onto Sabourad's Dextrose Agar supplemented with chloramphenicol and Mycosel Agar. Smooth, one-celled, subglobose to cylindrical arthroconidia quite variable in size, consistent with description of Geotrichum candidum. Histology findings of biopsied material stained with Hematoxylin and Eosin (HE) and Periodic Acid-Schiff (PAS), revealed the presence of numerous hyaline septate hyphae among necrotic tissues. Immediately after the clinical examination, in view of the extensive injuries, a subcutaneous single dose of 10,000 UI of benzathine penicillin G, a long therm-acting antibiotic, was administered and a daily topical application of 10% iodine was recommended. Supportive therapy was not necessary due to the animal general good health. The turtle was reexamined only 45 days later showing a little improvement of the lesions and new collected material yielded no bacteria but only pure colonies of G. candidum and still the presence of hyaline septate hyphae on the histopathologic assessment. Then, a treatment with topical nystatin was prescribed and 6 months later, the success of the therapy, was confirmed by the complete resolution of the wounds and the negative results of the third fungal and histological evaluations. Discussion: Geotrichosis has already been reported in animals, including lizards and snakes and reports about Geotrichum species as etiological agents among chelonians have been increasing in recent years. Although the conditions that enable opportunistic infections are similar for all saprobic fungi, clinical cases by Geotrichum species are rare in hosts. We believe that misreporting or misdiagnosis with other species, like Candida, could be the reason it is rarely reported. In this report, and in the other reviewed cases of animal geotrichosis, no predisposing factors are mentioned and the bite probably caused the wound in the shell and allowed the growth of the pathogenic fungus. The appropriate medium and the cultivation of Geotrichum candidum, as well as the use of molecular analysis, can contribute to the successful isolation of these species and make a correct final diagnosis.

This is the first description of cutaneous mucormycosis in buffalo in the Brazilian Amazon biome. All buffalo showed apathy, inappetence, weight loss, reluctance to move, and prolonged sternal decubitus. Of the four affected animals, two died 15 and 30 days after the appearance of clinical signs. In the initial phase, the skin lesions were rounded areas with dry central regions, sensitive to palpation, with protruding edges and diameters ranging from 8 cm to 15 cm. These areas of necrosis were isolated or coalescing and present mainly on the limbs and sides. In an advanced stage of the disease, there was detachment of the skin from the necrotic areas with extensive wound formation, which sometimes exposed the subcutaneous tissue. The histopathology of the skin showed a multifocal inflammatory infiltrate composed of intact and degenerated eosinophils surrounded by epithelioid macrophages. At the center of these areas was a focally extensive area of epidermal ulceration characterized by intact and degenerated neutrophils, the necrosis of epithelial cells, and the accumulation of fibrin and erythrocytes. The mycological culture was positive for Rhizopus sp. The diagnosis of cutaneous dermatitis caused by Rhizopus sp. was based on clinical signs, macroscopic and histopathological findings, and the identification of the fungus by mycological and molecular techniques.

Background: Dermatophytes, fungi of universal distribution, invade semi or fully keratinized structures, such as skin, fur/hair and nails. In companion animals (cats, dogs, or small mammals like rabbits, guinea pigs, and chinchillas) as well as in large animals (mainly in horses and cattle). Frequently are responsible for skin diseases including alopecia and crusts. This work reported a case of equine ringworm due to Nannizzia gypsea (Microsporum gypseum) detected from the clinical sample by SYBR-Green real-time PCR. The strategy was based on the DNA extraction directly from the infected hair followed by real-time PCR and melting-point analysis. Case: A 2-year-old horse was referred to the Veterinary Clinic Hospital, Federal University of Rio Grande do Sul (HCV-UFRGS), Porto Alegre, Brazil, presenting circular areas of alopecia and lesions with dry aspect and thin powdery scales and hairs broken at their base mainly on head and neck. No previous antifungal treatment was carried out. The sample was obtained by plucking the hair with forceps and scales from the peripheral area of the lesions. For mycological diagnosis, hair specimen was clarified and examined microscopically using 10% potassium hydroxide (KOH) for the visualization of arthroconidia (ectothrix type). The infected hair was plated onto MycoselTM Agar and Mycosel Agar with nicotinic acid requirement, incubated at 25-30 degrees C for 10-15 days. Microscopic features (macroconidia) and colony characteristics (colors and texture) were conducted for the differentiation of the species within the genus Microsporum. In addition, real-time PCR was applied for direct analysis of the fungal DNA obtained from the hair sample. Qiagen DNeasy((R)) plant mini DNA extraction kit protocol was used to extract DNA from the hair sample according to the manufacturer's instructions. A realtime PCR was performed using the pan-dermatophyte primers for detecting a DNA fragment encoding chitin synthase 1 using SYBR Green PCR Mix. The melting curve data were obtained by continuous fluorescence acquisition from 60 to 95 degrees C with a ramp rate of 0.3C. Microscopic examination of hair sample was negative. The culture was positive and dermatophyte present in the hair sample was confirmed as Nannizzia gypsea (M. gypseum) following the amplification of CHS1 gene. The hair sample melted at 83.78 degrees C, showing that the isolated clinical curve was distinct from the control (M. canis) melted at 85.3 degrees C. Discussion: Animals can be infected by a variety of dermatophytes. Nannizzia gypsea (Microsporum gypseum) is a geophilic keratinophilic fungus with a worldwide distribution which may cause infections in animals and humans, particularly children and rural workers during warm humid weather. Usually produces a single inflammatory skin or scalp lesion. The dermatophytic infection in horses is generally follicular and the most common clinical sign is one or many circular areas of alopecia with variable erythema, scaling and crusting. Is extremely important the culture of samples from skin lesions, because many agents may be involved and, frequently KHO test is negative. Conventional methods (direct exam and fungal culture) lacks the ability to make an early and specific diagnosis. The qPCR assay introduced in this study allows the specific detection of relevant dermatophytes in veterinary medicine in a short time. In the case reported here, dermatophytosis due Nannizzia gypsea (Microsporum gypseum) in a horse was confirmed based on mycological diagnosis and SYBR-Green real-time PCR.

ABSTRACT The aims of this study were to search for the presence of bacterial and fungal growth in urine samples from cats with urethral obstruction, to identify the causative infectious agents, and to compare select clinical-laboratory parameters with urine culture results after urethral catheterization. Twenty-eight male cats were enrolled in this prospective study. Urethral catheterization was performed aseptically, and the catheter was maintained for 24-48 hours with a sterile closed collection system. No antibiotics were used during this period. The first urine sample was obtained at the time of presentation, and the second, 24-48 hours after catheter removal, both by cystocentesis. Two cats (7.1%) presented a positive bacterial urine culture on the first sample. The frequency of positive urine cultures in the second sample was 52.4% (11/21 cats). One third of bacterial pathogens were Gram-positive, one third were Gram-negative, and one third were mixed isolates. Fungal cultures were all negative. There was significant association of positive urine culture results with pyuria (P=0.0128). A high frequency of positive urine cultures after urinary catheter removal was observed, despite respecting the standards of care for urethral catheters. Still, these results should be interpreted with caution, since intermittent bladder flushing was performed disconnecting the collecting system, which may have represented a critical point for bacterial contamination. It is emphasized that urine cultures should be considered as follow up in cats with urethral obstruction, after urethral catheter removal.

Lay Summary The aim of this work was to develop a molecular detection strategy for dermatophytes by SYBR-Green real-time PCR of hair samples from animals. The melting curve analysis of the CHS1 fragment revealed to be reproducible, showing a single distinct peak for distinct dermatophyte species and allowed the diagnosis of dermatophytosis in dogs and cats caused mainly by Trichophyton mentagrophytes, Microsporum sp., and Nannizzia gypsea). The classical dermatophytes diagnosis is based on mycological culture and microscopy observation both human and animal hair, skin, and nail samples. The aim of this work was to develop the new in-house real-time PCR with pan-dematophyte reaction for detection and identification of the main dermatophytes directly from hair samples, providing a simple and rapid diagnosis of dermatophytosis in dogs and cats. An in-house SYBR-Green real-time PCR was designed and used for detecting a DNA fragment encoding chitin synthase 1 (CHS1). A total of 287 samples were processed by culture, microscopic examination with KOH 10%, and real-time PCR (qPCR) analysis. Melting curve analysis of the CHS1 fragment revealed to be reproducible, showing a single distinct peak for each species of dermatophyte, namely Trichophyton mentagrophytes, T. verrucosum, Microsporum canis, and Nannizzia gypsea (formerly M. gypseum). Then, out of the 287 clinically suspected cases of dermatophytosis, 50% were positive for dermatophytes by qPCR, 44% by mycological culture, and 25% by microscopic examination. Microsporum canis was identified in 117 samples tested by culture and 134 samples tested by qPCR, followed by N. gypsea in 5 samples (either tested by culture or qPCR) and T. mentagrophytes detected in 4 and 5 samples when tested by culture or qPCR, respectively. Overall, qPCR allowed the diagnosis of dermatophytosis in clinical samples. The results suggest this newly proposed in-house real-time PCR assay can be used as alternative diagnosis and rapid identification of dermatophytes frequently associated to clinical hair samples of dogs and cats.

This study aimed to prospectively evaluate the risk factors of infection by Aelurostrongylus abstrusus in Brazilian cats with cough and/or radiographic changes, using as diagnostic tools the Baermann method (BM), polymerase chain reaction (PCR) of feces, bronchoalveolar lavage fluid (BALF), and cytology. Forty-three cats that were presented with cough or lung radiographic abnormalities compatible with bronchoalveolar disease were included in the study. After clinical evaluation, feces samples were collected to investigate lungworm parasitism through BM and PCR. BALF was performed to provide samples for cytology, bacteriology, and fungal culture. Stool PCR was considered the gold standard for diagnosis tests, and the other methods were evaluated by their agreement. PCR presented 74% (32/43) of positivity for A. abstrusus, while in the BM, 41% (18/43) were positive. BM showed sensitivity of 56.25% and specificity of 100% when compared with PCR. No larva was found in the cytological evaluation of 21 BALF samples. Lungworm is an important cause of bronchopulmonary disease in domestic cats in Brazil and should be included as a differential diagnosis when a cat is presented with cough or radiographic abnormalities. BM is a sensitive, non-invasive, and cheap technique to diagnose the disease, but it is not as sensitive as PCR.

Trichophyton verrucosum is the most commonly dermatophyte involved in cattle ringworm. This work reported a case of bovine dermatophytosis due to Trichophyton verrucosum detected from the clinical sample by SYBR-Green real-time PCR. The strategy was based on the DNA extraction directly from the infected hair followed by real-time PCR and melting-point analysis. A faster and differential diagnosis was observed when compared to the conventional mycological methodology for detection and identification of Trichophyton verrucosum.

Background: Dermatophytes, fungi of universal distribution, invade semi or fully keratinized structures, such as skin, fur/ hair and nails. The various species of dermatophytes are classified into three genera anamorphic: Microsporum, Trichophyton and Epidermophyton. The genus Epidermophyton includes only E. floccosum, that rarely affects animals. The main species responsible for the disease in dogs and cats are Microsporum canis, M. gypseum and Trichophyton mentagrophytes, which were characterized through conventional mycological methodology (microscopic examination with KOH and culture). Molecular methodologies, such as real-time PCR, can contribute to a rapid laboratory diagnosis, helping clinicians to initiate an early antifungal treatment. This case report describes a case of canine dermatophytosis due to Trichophyton mentagrophytes detected from a clinical sample by SYBR-Green real-time PCR. Case: A 8-year-old dog, rescued from the street, was referred to a private veterinary clinic in the city of Canoas, RS, Brazil, presenting generalized lymphadenomegaly, crusted lesions all over the body, generalized alopecia, signs of excoriation and epistaxis. Initially, were administered prednisone [1 mg/kg every 48 h, BID] and cephalexin [30 mg/kg, BID]. Weekly baths with benzoyl peroxide were also given. The therapy was not clinically successful. Wood's Lamp Test was negative. As a differential diagnosis, PCR for detection of Leishmania was negative. Complete blood count and serum biochemical assay were also performed. For mycological diagnosis, hair specimen was clarified and examined microscopically using 10% potassium hydroxide (KOH) for the visualization of chains of arthroconidia (ectothrix invasion of hair). The infected hair was plated onto MycoselTM Agar, incubated at 28 degrees C for 15 days. Microscopy of hyphae/ conidia and macroscopic colony characteristics (colors and texture) were conducted for the differentiation of the species within the genus Microsporum and Trichophyton. In addition, real-time PCR was applied for direct analysis of the fungal DNA obtained from the hair sample. Microscopic examination was negative. The dermatophyte present in the hair sample was confirmed as Trichophyton mentagrophytes by culture and qPCR (melting-point analysis). The patient was treated with systemic itraconazole [10 mg/ kg SID - 90 days]. Twice-weekly application of 2.5 % miconazole and 2% chlorhexidine shampoo until complete cure. Discussion: Dermatophytosis is often listed as self-limiting infection; however, animal dermatophytosis can spread between pets, as well as a zoonotic transmission to humans. The literature on dermatophytosis indicates that Microsporum canis is the predominant etiological agent, followed by M. gypseum. Trichophyon mentagrophytes that appear in a lower percentage of isolation. The culture of hair, even with specific medium containing chloramphenicol and cyclohexamide, may present contaminating fungi, not related to dermatophytosis, which can inhibit or override the growth of dermatophytes. The use of real-time PCR provided a faster and specific diagnosis of dermatophytosis when compared to the conventional mycological methodology for detection and identification of T. mentagrophytes, which takes around 10 to 15 days for culture. It is possible to use this technique as an alternative diagnosis for dermatophytes associated to clinical hair samples of dogs.

Sporotrichosis is a disease caused by thermally dimorphic fungi belonging to the Sporothrix schenckii complex. We report the occurrence of multiple cases of sporotrichosis in cats in Porto Alegre (RS), Brazil, from 2015 to 2019. A set of 21 fungal isolates were collected and the partial sequence of calmodulin (CAL) gene compared. All isolates were identified at phenotypic and molecular level as S. brasiliensis. Phylogenetic analysis showed the isolates clustered in two distinct groups, suggesting the presence of multiple genetic variants of S. brasiliensis in the region.

A ten year-old Holstein cow had an intermittent bloody diarrhea, evolving to anorexia and recumbency, followed by death. Mycotic segmental enteritis was diagnosed based on the pathological and immunohistochemical findings. Rhizopus microsporus was identified as the causal agent through fungal culture and PCR analysis. Intestinal mucormycosis is poorly described in cattle and should, therefore, be included as a differential diagnosis in cases of diarrhea and death in ruminants, especially when there is disruption of the normal balance of the alimentary microbiota.

This report describes a case of tracheobronchitis caused by Aspergillus fumigatus in a cow. A 4-year-old Jersey cow was submitted for necropsy examination. Grossly, the mucosa of the trachea and bronchi was diffusely reddened and was covered by a moderate amount of yellow, caseous material, sometimes with small foci of grey ‘cotton-like’ appearance, suggestive of filamentous fungus. Microscopical analysis of the trachea and bronchi showed marked diffuse mucosal and submucosal necrosis with a large amount of cellular debris, fibrinous exudation and inflammatory infiltration dominated by macrophages and neutrophils. There were large numbers of septate fungal hyphae with acute bifurcation angles and conidia and vesicles typical of Aspergillus spp. that were stained by the Grocott method. A. fumigatus growth in mycological culture from the trachea and bronchi was confirmed by molecular identification. Tracheobronchitis caused by A. fumigatus infection in cattle is poorly described and requires further research to improve the therapeutic management of the disorder.

Background: Cryptococcosis is a serious fungal infection contracted by humans and animals, and the most common systemic mycosis found in cats. This disease is often contracted through inhalation of fungal propagules. The Central Nervous System (CNS) may be infected through local extension (nasal and frontal sinuses) or via hematogenous route. Similarly to CNS bacterial infection, the clinical signs of neurological dysfunction may be attributed to mass effect (gelatinous mass of fungal microorganisms and fungal granuloma formation) or to a more disseminated inflammatory response to invading microorganisms. The objective of this study is to report one case of a patient with cryptococcal granulomas in the central nervous system and one case of a patient with neurological signs associated to cryptococcosis.Cases: A 3-year-old male mixed breed feline was admitted Veterinary Clinic, located in Porto Alegre, RS, Southern Brazil. The patient presented unsourced behavioral changes, vestibular ataxia and dysphagia caused by inability of coordination. The following tests were performed: complete blood count test, biochemical analysis, computed tomography scan (CT scan), fluid analysis, radiography and toxoplasmosis test. The following medicine were administrated for treatment: fluconazole, dexamethasone, mannitol, phenobarbital and levetiracetam. Fluid therapy was also part of the treatment. Immediately after death, the cat was submitted for necropsy, and a fungal granulomatous meningoencephalomyelitis was diagnosed. Cryptococcus sp. was identified as the causal agent through pathological findings, fungal culture and PCR analysis. One year later, another feline was admitted to the same clinic (a 2-year-old female mixed breed) presenting hypersalivation, tremors and excessive vocalization. The patient had contact with the deceased feline. The following tests were performed: complete blood count test, biochemical analysis, computed tomography scan (CT scan), cerebrospinal fluid analysis, and radiography. The following medicine were administrated for treatment: fluconazole, prednisolone, phenobarbital, potassium citrate and cefalotine. This patient is frequently monitored by a veterinarian and presents adequate health conditions after the occurrence. Cryptococcus sp. was identified as the causal agent through fungal culture and cytology (cerebrospinal fluid).Discussion: Cats are the most frequently infected animals with the involvement of the upper and or lower respiratory tract, subcutaneous granuloma, and disseminated infections. These animals present a higher quality of life when the disease is diagnosed and treated early. Peripheral enhancement of intracranial cryptococcal granulomas has been demonstrated in felines. These fungal granulomas often present evidence of significant perilesional edema. The most substantial evidence for diagnosis of cryptococcosis is the identification of the microorganism in Cerebrospinal fluid (CSF) samples. The mechanism of the lesion is cellular death, probably caused by secondary atrophy from distortion and tissue compression due to the expansion of Cryptococcus cysts in the cerebral parenchyma. There are several reports of long-lasting remission or cure of CNS cryptococcosis treated with medicine combinations, including flucytosine and/or triazole antifungal agents (itraconazole and fluconazole.) The isolated use of flucytosine may contribute to development of drug resistance. One controversial alternative that may be beneficial to the patient is the administration of low prednisolone doses to counter perilesional edema in the initial stages of treatment. The utilization of glucocorticoids after diagnosis was associated to higher survival rates in animals.

ABSTRACT: Susceptibility testing is essential to inform the correct management of Aspergillus infections. In this study we present antifungal susceptibility profile of A. fumigatus isolates recovered from lungs of birds with and without aspergillosis. Fifty three isolates were tested for their antifungal susceptibility to voriconazole (VRC), itraconazole (ITZ), amphotericin (AMB) and caspofungin (CSP) using the M38-A2 broth microdilution reference method. Five isolates were resistant to more than one antifungal drug (CSP + AMB, VRC + ITZ and AMB + ITZ). Fifteen (28%) isolates with susceptible increased exposure (I) to ITZ were sensible to VRC. Resistance to AMB (>2μg/mL) was observed in only four isolates. Eleven (21%) A. fumigatus present resistance to ITZ (13%) and VRC (8%). Fungal isolation from respiratory samples has been regarded as being of limited usefulness in the ante mortem diagnosis of aspergillosis in birds. However, the results suggest that the detection and antifungal susceptibility profile may be helpful for monitoring of therapy for avian species and where antifungal resistance might be emerging and what conditions are associated to the event.

Background: Deep fungal infections of the orbit and nasal passages causing rhinitis and ulcerative keratomycosis are uncommonly reported in cats. Hyalohyphomycetes and phaeohyphomycetes have rarely been associated with this disorder. Sino-orbital fungal diseases are emerging and more invasive than sino-nasal fungal diseases with poor response to therapy and a worse prognosis. Brachycephalic feline breeds seem to be at increased risk for development of upper respiratory fungal diseases. Diagnosis is based on the demonstration of fungal hyphae by cytology or histology and definitive confirmation by fungal culture and molecular methods. This is the first case report of a cat with clinical mixed fungal ball with Aspergillus and Scopulariopsis in Brazil.Case: A 3-year-old male Persian cat, in São José city, Santa Catarina, Brazil, was presented with exophthalmos and corneal ulcer of the left eye and protrusion, hyperemia, quemosis and fibroses of the left third eyelid. The retropulsion of the globe was negative in this eyeball and a presumptive diagnosis of a retrobulbar mass was made. The patient underwent a surgical procedure for inspection and collection of samples for bacterial and mycological culture. Culture revealed no bacterial growth, however, unique and abundant growth of Aspergillus spp. was present. A subconjunctival enucleation of the left eye was made and the mass was sent for histopathology examination. Histology showed inflammatory proliferative necrotizing pyogranulomatous reaction; with the presence of severe fungal infection evidenced by large number of hyaline septated regular and irregular mold hyphae. Molecular identification was performed using panfungal primers (ITS3-F / ITS4-R). Patient was treated with systemic itraconazole associated with amphotericin B and topical clotrimazole. A mass started to grow rapidly in the left pterygopalatine fossa and was surgically removed, but recurrence occurred seven days after. After 22 days of treatment, the animal died suddenly with a history of acute inspiratory dyspnea and cyanosis at the time prior to death. The diagnosis of sino-orbital fungal disease in the feline was based on clinical signs, mycological culture, histopathology and molecular methods.Discussion: Sino-orbital fungal diseases rare in cats and can result in significant injuries to the upper respiratory tract and eyes, sometimes resulting in enucleation and death. It seems feasible that a brachycephalic facial conformation may be an important risk factor for the development of sino-nasal fungal diseases in cats. Despite using selected drugs and eye enucleation to treat the disease, the cat developed a rapid growing oral mass that probably caused acute inspiratory dyspnea and death. Since no controlled studies exist on the treatment of feline fungal diseases, these cases are a challenge to the feline practitioner and this type of clinical manifestation should be included in the differential diagnosis of upper respiratory and ocular diseases.