Feline sporotrichosis due to Sporothrix globosa in the Metropolitan region of Buenos Aires: A case study

Abstract We describe here the first case of feline sporotrichosis caused by Sporothrix globosa, occurring outside the epizootic area of Buenos Aires province, Argentina. Unlike cases reported with Sporothrix brasiliensis, on this occasion there was no clinical or serological evidence of zoonotic transmission through scratches or bites from the sick cat to the attending veterinarian or the person responsible for its care. This report aimed to improve the knowledge about the pathogenic profile of S. globosa.

Resumen En este trabajo se describe el primer caso de esporotricosis felina causada por Sporothrix globosa fuera del área epizoótica de Buenos Aires. A diferencia de los casos documentados de Sporothrix brasiliensis, en este no hubo evidencia clínica ni serológica de transmisión zoonótica por arañazos o mordeduras del gato enfermo a su dueño ni al veterinario que lo atendió. Con este reporte se espera contribuir a un mejor conocimiento sobre el perfil patogénico de S. globosa.

Feline sporotrichosis due to Sporothrix globosa in the Metropolitan region of Buenos Aires: A case study.

We describe here the first case of feline sporotrichosis caused by Sporothrix globosa, occurring outside the epizootic area of Buenos Aires province, Argentina. Unlike cases reported with Sporothrix brasiliensis, on this occasion there was no clinical or serological evidence of zoonotic transmission through scratches or bites from the sick cat to the attending veterinarian or the person responsible for its care. This report aimed to improve the knowledge about the pathogenic profile of S. globosa.

Seroprevalence of sporotrichosis infection in cats living in the Buenos Aires epizootic area.

This study aimed to perform a seroepidemiological survey on the prevalence of sporotrichosis among cats living in the Northern area of Buenos Aires, where a four-fold increase of Sporothrix brasiliensis infections were diagnosed during the last decade. For this purpose, an in-house indirect ELISA test sensitized with S. brasiliensis crude antigens was used. The ELISA test showed 100.0% sensitivity and 95.0% specificity. Antibodies against S. brasiliensis antigens were detected in 3.7% (9/241) of healthy cats evaluated, suggesting likely exposure or infection to this fungus. This ELISA test would be a valuable screening tool for diagnosing sporotrichosis and for seroepidemiological surveys.

S. brasiliensis is the primary cause of feline sporotrichosis in Argentina. The seroprevalence of sporotrichosis infection in urban localities of Buenos Aires province is reported for the first time. An ELISA test using S. brasiliensis crude antigens is also described.

Feline microRNAome in ovary and testis: Exploration of in-silico miRNA-mRNA networks involved in gonadal function and cellular stress response.

The aim of the study was to perform the first in-depth analysis of miRNAs in ovarian and testicular tissues of the domestic cat, a critical biomedical model. Specifically, potential miRNA involvement was explored in gonadal function, testis development, and cellular stress response to preservation protocols. We performed miRNA-sequencing on 20 ovarian and 20 testicular samples from 15 cats, including different ages and tissue treatments. Using fresh tissues (n = 15), we confirmed gonadal expression of 183 miRNA precursors and discovered additional 52 novel feline candidate precursors. We integrated the mRNA data from our previous study on the same age and treatment groups to create in-silico miRNA-mRNA networks and their functional enrichment, which allows comprehensive exploration into possible miRNA functions in cat gonads. Clusters of miRNAs united by shared differentially expressed mRNA targets are potentially involved in testicular development and spermatogenesis. MicroRNAs could play a significant role in ovarian tissue response to stress from microwave-assisted dehydration, with smaller roles in cellular response to vitrification in both ovary and testis. This new list of miRNAs with potential function in cat gonads is a major step towards understanding the gonadal biology, as well as optimizing fertility preservation protocols.

Sporotrichosis caused by Sporothrix brasiliensis in Argentina: Case report, molecular identification and in vitro susceptibility pattern to antifungal drugs.

Sporotrichosis is considered a neglected disease of humans and animals in many regions of the world and is the most frequent implantation mycosis in Latin America. OBJECTIVES: To illustrate the zoonotic importance of the disease, describing a case involving a veterinarian and an infant that acquired the disease from a domestic cat and to describe, genotype and characterize these new isolates. METHODS: Direct examination of tissue samples from the two patients and feline lesions revealed the presence of Sporothrix yeast-like organisms. Fungal cultures and molecular identification of the strains were performed. Since antifungal susceptibility data of animal-borne isolates are scarce, the in vitro susceptibility testing by a microdilution reference method was determined against azoles, amphotericin B and terbinafine. RESULTS: Fungal culture and sequence analysis of the ITS region of rDNA and calmodulin and ß-tubulin genes confirmed the diagnosis and the causative agent as Sporothrix brasiliensis. In all cases, terbinafine was the most active drug, followed by posaconazole, itraconazole and voriconazole; the least active drugs were amphotericine B and fluconazole. Lack of clinical response in the veterinarian and in the infant to itraconazole and potassium iodide, respectively was observed. CONCLUSIONS: This study contributed to the molecular epidemiology of Sporothrix species in Argentina and the characterization of the in vitro susceptibility pattern of S. brasiliensis isolates recovered from a cat and two humans involved in this case of zoonotic sporotrichosis. Bearing in mind the "One Health" concept, the experience described in the present study highlights the need for future strategies for sporotrichosis treatment, control and prevention.

Domestic carnivore interactions with wildlife in the Cape Horn Biosphere Reserve, Chile: husbandry and perceptions of impact from a community perspective.

BACKGROUND: Hundreds of millions of domestic carnivores worldwide have diverse positive affiliations with humans, but can provoke serious socio-ecological impacts when free-roaming. Unconfined dogs (Canis familiaris) and cats (Felis catus) interact with wildlife as predators, competitors, and disease-transmitters; their access to wildlife depends on husbandry, perceptions, attitudes, and behaviors of pet owners and non-owners. METHODS: To better understand husbandry and perceptions of impacts by unconfined, domestic carnivores, we administered questionnaires (n = 244) to pet owners and non-owners living in one of the last wilderness areas of the world, the Cape Horn Biosphere Reserve, located in southern Chile. We used descriptive statistics to provide demographic pet and husbandry information, quantify free-roaming dogs and cats, map their sightings in nature, and report experiences and perceptions of the impact of free-roaming dogs and cats on wildlife. We corroborated our results with an analysis of prey remains in dog feces (n = 53). With generalized linear models, we examined which factors (i.e., food provisioning, reproductive state, rural/village households, sex, and size) predicted that owned dogs and cats bring wildlife prey home. RESULTS: Thirty-one percent of village dogs (n = 121) and 60% of dogs in rural areas (n = 47) roamed freely day and/or night. Free-roaming dog packs were frequently observed (64% of participants) in the wild, including a feral dog population on Navarino Island. Dogs (31 of 168) brought home invasive muskrats (Ondatra zibethicus) and avian prey, and over half of all cats (27 of 51) brought home mainly avian prey. Birds were also the most harassed wildlife category, affected by one third of all dogs and cats. Nevertheless, dog-wildlife conflicts were hardly recognized (<9% of observed conflicts and suspected problems), and only 34% of the participants thought that cats might impact birds. Diet analysis revealed that dogs consumed livestock (64% of 59 prey occurrences), beavers (Castor canadensis, 14%), and birds (10%). The probability that dogs brought prey to owners' homes was higher in rural locations and with larger dogs. There was also evidence that cats from rural households and with an inadequate food supply brought more prey home than village cats. DISCUSSION: Although muskrat, beavers, and birds were brought home, harassed, or found in dog feces, free-roaming dogs and, to a lesser extent, cats are perceived predominantly in an anthropogenic context (i.e., as pets) and not as carnivores interacting with wildlife. Therefore, technical and legal measures should be applied to encourage neutering, increase confinement, particularly in rural areas, and stimulate social change via environmental education that draws attention to the possibility and consequences of unconfined pet interaction with wildlife in the southernmost protected forest ecoregion of the globe.

Conservation of spermatogonial stem cell marker expression in undifferentiated felid spermatogonia.

Spermatogonial stem cells (SSCs) are distinct in their ability to self-renew, transmit genetic information, and persist throughout the life of an individual. These characteristics make SSCs a useful tool for addressing diverse challenges such as efficient transgenic production in nonrodent, biomedical animal models, or preservation of the male genome for species in which survival of frozen-thawed sperm is low. A requisite first step to access this technology in felids is the establishment of molecular markers. This study was designed to evaluate, in the domestic cat (Felis catus), the expression both in situ and following enrichment in vitro of six genes (GFRA1, GPR125, ZBTB16, POU5F1, THY1, and UCHL1) that had been previously identified as SSC markers in other species. Antibodies for surface markers glial cell line-derived neurotrophic factor family receptor alpha 1, G protein-coupled receptor 125, and thymus cell antigen 1 could not be validated, whereas Western blot analysis of prepubertal, peripubertal, and adult cat testis confirmed protein expression for the intracellular markers ubiquitin carboxy-terminal hydrolase 1, zinc finger and BTB domain-containing protein 16, and POU domain, class 5, transcription factor 1. Colocalization of the markers by immunohistochemistry revealed that several cells within the subpopulation adjacent to the basement membrane of the seminiferous tubules and identified morphologically as spermatogonia, expressed all three intracellular markers. Studies performed on cheetah (Acinonyx jubatus) and Amur leopard (Panthera pardus orientalis) testis exhibited a conserved expression pattern in protein molecular weights, relative abundance, and localization of positive cells within the testis. The expression of the three intracellular SSC marker proteins in domestic and wild cat testes confirms conservation of these markers in felids. Enrichment of marker transcripts after differential plating was also observed. These markers will facilitate further studies in cell enrichment and IVC of felid SSCs enabling both production of transgenic domestic cats and preservation of the male genome from rare and endangered felids.

Experimental transmission of Cystoisospora felis-like coccidium from bobcat (Lynx rufus) to the domestic cat (Felis catus).

Cystoisospora felis is an ubiquitous coccidian of cats. The domestic cat (Felis catus) is its definitive host and several mammalian and avian species are its optional intermediate/transport hosts. Nothing is known if it is transmissible to wild felids. In the present study C. felis-like oocysts were found in two naturally infected bobcats (Lynx rufus) from Pennsylvania. To study transmission of C. felis-like parasite from bobcats to domestic cats, sporulated oocysts of C. felis-like from one bobcat were orally inoculated into interferon gamma gene knockout (KO) mice, and 56 days later tissues of KO mice were fed to two coccidian-free cats; two littermate cats were uninoculated controls. The inoculated cats and controls were euthanized five and seven days later, and their small intestines were studied histologically. One inoculated cat excreted C. felis-like oocysts seven days post inoculation (p.i.) and was immediately euthanized. Mature schizonts, mature male and female gamonts, and unsporulated oocysts were found in the lamina propria of small intestine; these stages were morphologically similar to C. felis of domestic cats. No parasites were seen in histological sections of small intestines of the remaining three cats. The experiment was terminated at seven days p.i. (minimum prepatent period for C. felis) to minimize spread of this highly infectious parasite to other cats. Although oocysts of the parasite in bobcats were morphologically similar to C. felis of domestic cats, the endogenous stages differed in their location of development. The bobcat derived parasite was located in the lamina propria of ileum whereas all endogenous stages of C. felis of domestic cats are always located in enterocytes of intestinal epithelium. Characterization of DNA isolated from C. felis-like oocysts from the donor bobcat revealed that sequences of the ITS1 region was only 87% similar to the ITS1 region of C. felis from domestic cats. These results indicate that the parasite in bobcat is likely different than C. felis of cats.

Enzyme immunoassays as a method for quantifying hair reproductive hormones in two felid species.

Non-invasive monitoring of wild felid reproductive states is important, given that many species reproduce poorly in captivity. Despite extensive work in faecal hormone analysis in felids, continued development of techniques is necessary, particularly with wild populations. In this study, our aims were as follows: (i) biochemical validation of enzyme immunoassays for estrogen, testosterone and progesterone in Canada lynx and domestic cat hair extracts; (ii) assessment of the use of hair reproductive hormones to differentiate between reproductive states (intact, estrus, pregnant and spayed/neutered), using domestic cats as a model; and (iii) assessment of the use of hair reproductive hormones to differentiate between age and sex, accounting for potential regional variability in wild lynx populations. Analysis of hair hormone levels showed prospective value in detecting pregnancy states, with pregnant domestic cats having higher levels of progesterone than spayed females. However, intact and pregnant cats did not differ in progesterone levels. Yet, two female domestic cats had higher levels of hair progesterone following a 38-day oral progestin treatment, perhaps providing a preliminary pharmacological validation of the method. Estrogen and testosterone did not differ statistically according to reproductive states of domestic cats, although intact males had higher levels of hair testosterone than neutered males. When we applied these techniques to lynx fur, we determined that hormone levels were not sufficiently precise to differentiate age classes. Hair reproductive hormone ratios differed between sexes, with the estrogen-to-progesterone ratio demonstrating the highest accuracy in differentiating males from females. Hair hormone levels differed regionally for wild lynx, indicating that spatial variability should be a consideration in wildlife hormone studies spanning large spatial scales. We conclude that use of hair hormone analysis by enzyme immunoassay may hold promise for differentiating sex in felids, but the technique will require further refinement and validation before it can be applied broadly and reliably.