CHARACTERIZATION OF THE FUNGAL MICROBIOTA IN THE NOSTRILS AND RECTUM OF AMAZONIAN MANATEES (TRICHECHUS INUNGUIS) FROM A REHABILITATION PROGRAM IN BRAZIL.

The present study characterized the filamentous and yeast-like fungal microbiota of the nasal cavity and rectum of Amazonian manatees (Trichechus inunguis) undergoing rehabilitation at the Laboratory of Aquatic Mammals, National Institute of Amazonian Research, Manaus, Amazonas, and determined the antifungal susceptibility of these organisms. Nasal and rectal swabs were collected from 22 calves and three juveniles. The samples were seeded in Sabouraud agar supplemented with chloramphenicol 10%, incubated at 26°C, and observed daily for up to 7 d. The growth of different filamentous and yeast-like fungi was observed among the two anatomical sites. Filamentous fungi were categorized by macro- and microscopic characteristics of the colonies. Representatives of each group were selected for molecular identification based on the internal transcribed spacer region. Yeast identification was performed using MALDI-TOF MS and molecular analyses. Thirteen genera of filamentous fungi and six genera of yeasts were isolated and identified. The dominant filamentous species were Fusarium spp., Aspergillus spp., and Cochliobolus lunatus in the nostril samples and Aspergillus melleus in the rectal samples. Candida was the dominant genus among the identified yeasts at both anatomical sites. In the antifungal susceptibility test, 28 isolates showed resistance to fluconazole (78%), itraconazole (39%), and nystatin (42%). The knowledge of fungal microbiota composition of Amazonian manatees provides information that assists in monitoring the health status of individuals maintained in captivity, as these organisms can behave either as opportunists or as primary pathogens. Moreover, the composition and resistance of these organisms may vary among different rehabilitation institutions or different time frames of search, reinforcing the importance of constant in loco surveillance of these microorganisms. This study provides new perspectives on the fungal diversity in the microbiota of manatees and supports future studies concerning the clinical and epidemiological aspects and the impacts of these agents on the health of Amazonian manatees undergoing rehabilitation.

Emergence of zoonotic sporotrichosis due to Sporothrix brasiliensis in Minas Gerais, Brazil: A molecular approach to the current animal disease.

Sporotrichosis is a neglected fungal zoonosis with significant impacts on human and animal health. Accurate diagnosis, treatment, and understanding of the transmission dynamics of Sporothrix species are essential for mitigating the spread of sporotrichosis. This study aimed to identify the Sporothrix species involved in the ongoing outbreaks of animal sporotrichosis in the metropolitan region of Belo Horizonte, Minas Gerais, Brazil, and analyse the phylogenetic relationships between pathogenic species to investigate the outbreak origin. Additionally, to better understand the evolution of the disease, we conducted a retrospective survey of positive feline and canine cases from November 2017 to July 2021 with proven cultures for Sporothrix. A significant increase in animal cases over the last 4 years was observed, with cats being the most affected host. Sporothrix brasiliensis was the predominant agent in 100% of the clinical isolates (n = 180) molecularly identified. Phylogenetic and haplotype analysis points towards the cases isolated from Minas Gerais sharing the haplotype originating from a long-lasting outbreak of cat-transmitted sporotrichosis in Rio de Janeiro, however, with a secondary contribution from genotypes circulating in other outbreaks in Brazil. Thus, we present clear evidence of the circulation of different S. brasiliensis genotypes associated with animal sporotrichosis in the metropolitan region of Belo Horizonte. Genetic monitoring can contribute to understanding the causal agent for zoonotic sporotrichosis in epidemiological processes and help to implement disease prevention and control measures.

The Antimicrobial Applications of Nanoparticles in Veterinary Medicine: A Comprehensive Review.

Nanoparticles (NPs) are nanoscaled particles sized from 1-100 nm, which can be composed of inorganic or organic compounds. NPs have distinctive morphology, size, structure, and surface features, which give them specific properties. These particular attributes make them interesting for biological and medical applications. Due to these characteristics, researchers are studying the possible aptness of numerous nanoparticles in veterinary medicine, such as the capacity to act as a drug delivery system. The use of these NPs as a possible bactericidal or bacteriostatic medication has been studied against different bacteria, especially multiresistant strains and the ones that cause mastitis disease. The antibiofilm property of these nanostructures has also already been proved. The antiviral activity has also been shown for some important viral animal diseases; the antifungal activity had been demonstrated against both pathogenic and mycotoxigenic species. Therefore, this review aimed to elucidate the main clinical and preventive veterinary applications of inorganic and organic nanoparticles.

Inhibitory Effect of GRAS Essential Oils and Plant Extracts on the Growth of Aspergillus westerdijkiae and Aspergillus carbonarius Strains.

The effect of essential oils (obtained using hydrodistillation) and plant extracts (ethanolic, aqueous, and hexanic extractions) of 10 different plants cultivated in Brazil were tested using the diffusion agar method, with the objective of evaluating the inhibitory effect of the oils and extracts on the mycelial growth of Aspergillus westerdijkiae NRRL 3174 and A. carbonarius RC 2054 (UNRC). Of the 40 essential oils and plant extracts analyzed, oregano essential oil and plant extract, rosemary essential oil, and the clove ethanolic extract were the best choice to obtain the growth parameters (radial growth rates (mm day-1) and lag phase (h)) due the good results presented and the volume of oil/extract obtained. Comparing all the essential oils and plant extracts that were tested for growth parameters, the best results were obtained for the clove ethanolic extract for both strains assayed. These results demonstrated an outstanding potential use of some of these products in prevention of fungal contamination in food. However, further studies need to be conducted to determine the ability of these oils and extracts to inhibit or reduce ochratoxin A production.

Mycotoxins in Cattle Feed and Feed Ingredients in Brazil: A Five-Year Survey.

Mycotoxins are toxic secondary metabolites produced by a variety of fungi, which when ingested can cause several deleterious effects to the health of humans and animals. In this work, the detection and quantification of six major mycotoxins (aflatoxins-AFLA, deoxynivalenol-DON, fumonisins-FUMO, ochratoxin A-OTA, T-2 toxin-T-2 and zearalenone-ZON) in 1749 samples of feed and feed ingredients for cattle, collected in Brazil between 2017 and 2021, was carried out using enzyme-linked immunosorbent assay (ELISA). In total, 97% of samples were contaminated with at least one mycotoxin, yet, very few samples exceeded the lowest European Union guidance values for cattle, and the estimated daily intake also showed a low risk for the animals. However, co-occurrences were widely observed, as 87% of samples contained two or more mycotoxins at the same time, and the presence of more than one mycotoxin at the same time in feed can lead to interactions. In conclusion, the contamination of feed and feed ingredients for cattle with mycotoxins in Brazil is very common. Hence, the monitoring of these mycotoxins is of significant importance for food safety.

Household outbreak of sporotrichosis: towards the One Health approach.

Although sporotrichosis requires a broad approach for control, few reports have described the relationship between the index case and secondary contacts. In the present work, we report an outbreak involving a woman, a dog, and two cats from the same household environment, including the clinical and epidemiological aspects and outcomes, and discuss the importance of a One Health approach to face this neglected disease. The joint efforts of professionals such as veterinarians and physicians are essential for early diagnosis and surveillance, which contributes to the rapid identification and control of zoonotic sporotrichosis outbreaks.

Household outbreak of sporotrichosis: towards the One Health approach

ABSTRACT Although sporotrichosis requires a broad approach for control, few reports have described the relationship between the index case and secondary contacts. In the present work, we report an outbreak involving a woman, a dog, and two cats from the same household environment, including the clinical and epidemiological aspects and outcomes, and discuss the importance of a One Health approach to face this neglected disease. The joint efforts of professionals such as veterinarians and physicians are essential for early diagnosis and surveillance, which contributes to the rapid identification and control of zoonotic sporotrichosis outbreaks.

Characterization of animal sporotrichosis in a highly urbanized area.

An epidemiological characterization of animal sporotrichosis was carried out between 2017 and 2018, in a highly urbanized area in Brazil, including outcomes and the spatial distribution of the cases according to the health vulnerability index (HVI) of the study territory. One hundred and sixty-five cats and four dogs suspected of sporotrichosis were identified by a surveillance and control program previously implemented in the study area. One hundred and five of these animals (62.1 %) were considered positive for Sporothrix spp., of which 103 were cats and two were dogs. Cases predominated among male cats (53.4 %), although gender did not present a statistically significant association (p > 0.05) with sporotrichosis cases. Considering the positive cats, 55.4 % were not neutered and free access to the street was reported as a habit for 86.4 % of them. Roaming free in the streets increased the chance of infection by 2.54 times. We observed a high frequency of euthanasia or death (48.8 %) among 86 infected cats available to be included in the follow-up stage of the study, even when they were treated (46.5 %) and a low cure rate (31 %). The disease spread, unrelated to the HVI in the territory. Data produced suggested that avoiding access to the street seems to have greater importance to sporotrichosis control in cats than neutering and reinforces the importance of health education, especially in relation to responsible feline ownership. The free offer of diagnosis and treatment also should be taken into consideration as important measures to control the disease.

Implementation of an Animal Sporotrichosis Surveillance and Control Program, Southeastern Brazil.

We report the implementation of an animal sporotrichosis surveillance and control program that evaluates strategies to identify suspected and infected cats in a municipality in southeastern Brazil. All adopted measures reinforced the program, although strategies had different abilities to detect the presence of infection.

Spatial association between sporotrichosis in cats and in human during a Brazilian epidemics.

Sporotrichosis is a subcutaneous granulomatous disease caused by the fungus Sporothrix spp. In Brazil, S. brasiliensis is reported in regions of outbreaks and epidemics in the zoonotic form of the disease where cats play an important role in the transmission of the disease to humans. Therefore, it is important to assess how the presence of infected cats impacts the risk for sporotrichosis in humans. The objective of this study was to analyze the spatial association of sporotrichosis in cats and in humans from Belo Horizonte, a Brazilian city where an epidemics of sporotrichosis occurs since the first human case register in 2015, through an inhomogeneous Poisson process model. Feline and human cases of sporotrichosis recorded between January 2016 and June 2019 were georeferenced by address and spatial point patterns were generated. Feline case intensity and human demographic density were calculated using a kernel smoothed estimate. The distance to the nearest feline case was also compute. Model parameters were estimated by Maximum Likelihood Estimate. The model validation was performed by the evaluation of partial residual, leverage and influence measure. There were 343 cases of cats and 135 human cases of sporotrichosis. The average incidence of human sporotrichosis in the period was 1.343 per 100 thousand inhabitants, which is relatively low in relation to the population, but higher than that observed in other regions in zoonotic outbreak of the disease. The southern region of the municipality has a higher intensity of feline cases. According to the fitted model, the risk for human sporotrichosis is greater when at distances very close to a feline case, with a virtually stable effect for distances greater than 1 km. Regarding the intensity of feline cases there is a gradual increase in risk as the intensity of cases increases. From the leverage analysis it was observed that the model was particularly sensitive to the occurrence of human cases in the south and east regions, places with extreme values ​​of covariates. Poisson point process model seems to be a reasonable approach in spatial epidemiology when multiple sources of infection are involved, and there is a low incidence of the disease as long as it is reasonable to assume independence between cases. Interventions for disease prevention and control in humans are suggested to encompass disease control in cats and the search for feline cases, focused on diagnosis and control, close to reported human cases.

Associated factors and spatial patterns of the epidemic sporotrichosis in a high density human populated area: A cross-sectional study from 2016 to 2018.

An epidemiological characterization of human and feline sporotrichosis was carried out between 2016 and 2018, in a high density-populated area in Brazil. Professionals were trained to identify suspect cats and notify vets to interview the owners and collect swabs of the wounds from these animals. Mycological cultures were performed, and colonies identified as Sporothrix spp. Subsequently, data regarding the outcome from suspect animals were collected. Confirmed cases of human sporotrichosis (56) were also counted and analyzed for spatial distribution. 118 positive animals were observed. The prevalence of feline sporotrichosis was 8.36 ‰ (CI 95 %, 5.38-9.55 ‰). The odds for being positive in animals that lived only partially at home were 3.02 times greater than for those cats without access to the street (OR 3.02, CI 95 % 1,96-10,43). There was no statistically significant association between environmental variables and positive diagnosis, corroborating the hypothesis that direct transmission by infected cats plays a greater role in the occurrence and continuous outbreaks of sporotrichosis in Brazil. Among the positive animals, 61.90 % (CI 95 % 58.95-64.96) died, and they had an odds to die in the next six months 6.30 times greater than negative animals (p < 0.05, OR 6.30, CI 95 % 2,79-14,42). The case fatality rate was 55.08 % in cats (CI 95 % 49.20-51.15). The cause-specific death rate was 4.6 ‰ in cats (CI 95 % 3.4-6 ‰). Only 7.62 % (CI 95 % 7.12-8.16) positive cats were treated and cured. Among dead positive animals, 29.23 % were inappropriately discarded. In the study period 56 human cases were recorded in the Barreiro region. Regions with highest prevalence of feline sporotrichosis, had greater frequencies of both human and feline cases. This is the first report on the epidemic of sporotrichosis in Minas Gerais, Brazil. The free offer for treatment and veterinary care for these animals should be taken into consideration, as well as the collection and incineration of the dead ones, as measures of public health, followed by the guidance and care for the human patient.

In Vitro Activity of Neem (Azadirachta indica) Oil on Growth and Ochratoxin A Production by Aspergillus carbonarius Isolates.

Aspergillus carbonarius is a saprobic filamentous fungus, food spoiling fungus and a producer of ochratoxin A (OTA) mycotoxin. In this study, the in vitro antifungal activity of neem oil (0.12% p/p of azadirachtin) was evaluated against the growth of six strains of A. carbonarius and the production of OTA. Four different concentrations of neem oil were tested in addition to three incubation times. Only the concentration of 0.3% of neem oil inhibited more than 95% of the strain's growth (97.6% ± 0.5%), while the use of 0.5% and 1.0% of neem oil showed lower antifungal activity, 40.2% ± 3.1 and 64.7% ± 1.1, respectively. There was a complete inhibition of OTA production with 0.1% and 0.3% neem oil in the four strains isolated in the laboratory from grapes. The present study shows that neem essential oil can be further evaluated as an auxiliary method for the reduction of mycelial growth and OTA production.

Administração seriada de salina hipertônica 7, 5% na terapia para sepse grave decorrente da síndrome da diarreia hemorrágica aguda em cães / Serial administration of 7. 5% hypertonic saline solution in the therapy of severe sepsis due to acute hemorrhagic diarrhea syndrome in dogs

Na clínica de animais de companhia é frequente cães com síndrome da diarreia hemorrágica aguda associada a quadros de sepse, o que acarreta alta mortalidade. Nesse contexto, objetivou-se, em um ensaio clínico controlado aleatorizado de centro único, estudar os efeitos tardios da solução salina hipertônica a 7,5% em aplicações seriadas, sobre variáveis hemodinâmicas, clínicas e laboratoriais em cães com quadro de sepse grave decorrente desta síndrome. Para tal, 12 cães foram aleatoriamente distribuídos em dois grupos de igual número, sendo um controle (CON) e o outro, solução salina hipertônica 7,5% (SSH). Variáveis clínicas e laboratoriais foram avaliadas imediatamente após a admissão do paciente (T0), 24 (T24), 48 (T48) e 72 (T72) horas após a admissão. O grupo SSH recebeu Ringer com lactato, antibioticoterapia, analgésico e SSH 7,5% em bolus (5mL kg-1 em 4 minutos) no T24 e no T48. O grupo CON recebeu a mesma terapia acima, porém ao invés da utilização de SSH a 7,5%, administrou-se bolus de solução de Ringer lactato na mesma dose e tempos utilizado. As avaliações em cada tempo foram realizadas anteriormente à administração dos bolus, nos dois grupos. Avaliaram-se hemograma completo e as variáveis clínicas (escala AVDN, grau de desidratação, frequência respiratória e temperatura retal), frequência cardíaca (FC), pressão arterial sistólica (PAS). Os dados paramétricos foram avaliados pelos testes Student Newman Keuls e teste t de Student, e os não paramétricos pelo teste de Friedman e Mann Whitney, com nível de significância de P≥0,05. Nas variáveis clínicas estudadas não se observou diferença entre os grupos e entre os momentos avaliados. Observou-se diferença significativa no grupo SSH no T72, com elevação da PAS e redução da FC, fato não observado no grupo CON, onde esses parâmetros não se alteraram. O hematócrito e a concentração de hemoglobina diminuíram em ambos os grupos com o tempo. As contagens dos leucócitos totais e dos monócitos apresentaram uma elevação significativa no grupo SSH, estando os leucócitos dentro da faixa de normalidade no T72. Não houve diferenças significativas em relação aos neutrófilos segmentados, porém no grupo SSH verificou-se aumento de 9,5 vezes no T72 comparado com o T24 (P=0,09), enquanto que este aumento foi de apenas 2,5 vezes no grupo CON (P=0,30). Observou-se ainda redução nas contagens de plaquetas e na concentração de globulinas no grupo COM, enquanto essas variáveis se mantiveram estáveis no grupo SSH. Conclui-se que a administração seriada de SSH 7,5% se mostrou promissora no tratamento de cães com síndrome da diarreia hemorrágica aguda, pois auxilia na estabilização dos leucócitos, plaquetas e globulinas de cães com sepse grave decorrente da síndrome da diarreia hemorrágica aguda.(AU)

The association between acute hemorrhagic diarrhea syndrome and sepsis is frequent in dogs and causes high mortality. In this context we investigated in a randomized single-center controlled trial the late effects of 7.5% hypertonic saline solution in serial applications on hemodynamic, clinical and laboratory variables in dogs with severe sepsis due to the syndrome. Twelve dogs were randomly distributed into two groups of equal numbers, control (CON) and 7.5% hypertonic saline solution (SSH). Clinical and laboratory variables were evaluated immediately after admission of patients (T0), 24 (T24), 48 (T48) and 72 (T72) hours after the admission. The SSH group received Ringer with lactate, antibiotic therapy, analgesic and 5mL/kg-1 bolus of 7.5% SSH over 4 minutes on T24 and T48. The CON group received the same therapy, but instead of 7.5% SSH, Ringer with lactate bolus was given at the same dosis and times. All evaluations were performed prior to the administration of the bolus in both groups. Data collection included complete blood count and clinical variables (AVDN scale, degree of dehydration, respiratory rate and rectal temperature), heart rate (HR) and systolic blood pressure (SBP). Parametric data were evaluated by the Student Newman Keuls and Student t tests, and the nonparametric ones by the Friedman and Mann Whitney test, with a significance level of 0.05. There were no significant differences between the groups and moments in clinical variables. Nevertheless, the SSH group presented a significant elevation of SBP and HR reduction at T72. Hematocrit and hemoglobin concentration decreased in both groups over time. Total leukocyte and monocyte counts showed a significant elevation in the SSH group, however the leukocytes were within the normal range at T72. There were no significant differences in the segmental neutrophils, but a 9.5-fold increase in T72 compared to T24 (P=0.09) in the SSH group was observed, whereas this increase was only 2.5-fold in the CON group (P=0.30). A decrease in platelet counts and globulin concentration was observed in the CON group, while these variables remained stable in the SSH group. In conclusion, serial administration of 7.5% SSH is promising in the therapy of dogs with acute hemorrhagic diarrhea syndrome, since it assists in the stabilization of leukocytes, platelets and globulins in dogs with severe sepsis due to this syndrome.(AU)

Bakery by-products based feeds borne-Saccharomyces cerevisiae strains with probiotic and antimycotoxin effects plus antibiotic resistance properties for use in animal production.

The aim of this study was to select S. cerevisiae strains able to exert probiotic and antimycotoxin effects plus antibiotics resistance properties for use in animal production. S. cerevisiae LL74 and S. cerevisiae LL83 were isolated from bakery by-products intended for use in animal feed and examined for phenotypic characteristics and nutritional profile. Resistance to antibiotic, tolerance to gastrointestinal conditions, autoaggregation and coaggregation assay, antagonism to animal pathogens and aflatoxin B1 binding were studied. S. cerevisiae LL74 and S. cerevisiae LL83 showed resistance to all the antibiotics assayed (ampicillin, streptomycin, neomycin, norfloxacin, penicillin G, sulfonamide and trimethoprim). The analysis showed that exposure time to acid pH had a significant impact onto the viable cell counts onto both yeast strains. Presence of bile 0.5% increased significantly the growth of the both yeast strains. Moreover, they were able to tolerate the simulated gastrointestinal conditions assayed. In general, the coaggregation was positive whereas the autoaggregation capacity was not observed. Both strains were able to adsorb AFB1. In conclusion, selected S. cerevisiae LL74 and S. cerevisiae LL83 have potential application to be used as a biological method in animal feed as antibiotic therapy replacement in, reducing the adverse effects of AFB1 and giving probiotic properties.