Frequent Infection of Cats With SARS-CoV-2 Irrespective of Pre-Existing Enzootic Coronavirus Immunity, Brazil 2020.

Carnivores such as cats and minks are highly susceptible to SARS-CoV-2. Brazil is a global COVID-19 hot spot and several cases of human-to-cat transmission have been documented. We investigated the spread of SARS-CoV-2 by testing 547 domestic cats sampled between July-November 2020 from seven states in southern, southeastern, and northeastern Brazil. Moreover, we investigated whether immune responses elicited by enzootic coronaviruses affect SARS-CoV-2 infection in cats. We found infection with significantly higher neutralizing antibody titers against the Gamma variant of concern, endemic in Brazil during 2020, than against an early SARS-CoV-2 B.1 isolate (p<0.0001), validating the use of Gamma for further testing. The overall SARS-CoV-2 seroprevalence in Brazilian cats during late 2020 validated by plaque reduction neutralization test (PRNT90) was 7.3% (95% CI, 5.3-9.8). There was no significant difference in SARS-CoV-2 seroprevalence in cats between Brazilian states, suggesting homogeneous infection levels ranging from 4.6% (95% CI, 2.2-8.4) to 11.4% (95% CI, 6.7-17.4; p=0.4438). Seroprevalence of the prototypic cat coronavirus Feline coronavirus (FCoV) in a PRNT90 was high at 33.3% (95% CI, 24.9-42.5) and seroprevalence of Bovine coronavirus (BCoV) was low at 1.7% (95% CI, 0.2-5.9) in a PRNT90. Neutralizing antibody titers were significantly lower for FCoV than for SARS-CoV-2 (p=0.0001), consistent with relatively more recent infection of cats with SARS-CoV-2. Neither the magnitude of SARS-CoV-2 antibody titers (p=0.6390), nor SARS-CoV-2 infection status were affected by FCoV serostatus (p=0.8863). Our data suggest that pre-existing immunity against enzootic coronaviruses neither prevents, nor enhances SARS-CoV-2 infection in cats. High SARS-CoV-2 seroprevalence already during the first year of the pandemic substantiates frequent infection of domestic cats and raises concerns on potential SARS-CoV-2 mutations escaping human immunity upon spillback.

Retrospective surveillance of severe acute respiratory syndrome coronavirus 2 in pets from Brazil.

BACKGROUND AND AIM: The emerging concerns regarding the new Coronavirus's ability to cause infection in pets has led to animal testing and worrisome findings reported all over the world in domesticated and wild animals. This study aimed to investigate severe acute respiratory syndrome coronavirus (SARS-CoV)-2 by quantitative reverse transcription-polymerase chain reaction in dog and cat samples with the clinical presentation for respiratory or gastrointestinal disease in Brazil. MATERIALS AND METHODS: One hundred and twenty-five samples were collected from 12 states of Brazil that originated from the gastrointestinal, upper respiratory tract, and other sites, including some pools of samples from before the onset of the pandemic including blood and/or urine samples. They were tested for RT-PCR detection of respiratory or gastrointestinal pathogens through Respiratory or Diarrhea RT-PCR Panels in the TECSA (Tecnologia em Saninade Animal - Animal Health Technology) Veterinary Medicine Laboratory. This work was conducted in compliance with ethical standards. RESULTS: Seven different microorganisms that can cause respiratory and/or gastrointestinal clinical signs were detected in cats (Feline Coronavirus [FCoV], Feline Parvovirus, Feline Leukemia Virus, Feline Calicivirus, Mycoplasma felis, Campylobacter spp., and Cryptosporidium spp.) and three in dogs (canine distemper virus, Cryptosporidium spp., and Babesia spp.). CONCLUSION: Although the samples corresponded to the beginning of coronavirus disease-19 spread in Brazil and clinically correlated with the expected viral replication sites, none of the animals tested positive for SARS-CoV-2; reassuringly, four cats tested positive or FCoV none of them were positive for SARS-CoV2. The epidemiological surveillance of SARS-CoV-2 in pets is considered a one health issue, important for monitoring the disease evolution, spread and minimizing the animal-human health impacts, and directing Public Health Policies.

Canine Olfactory Detection of SARS-COV2-Infected Patients: A One Health Approach.

The aim of the present study is to apply the canine olfactory sensitivity to detect COVID-19-positive axillary sweat samples as a One Health approach in Latin America. One hundred volunteers with COVID-like symptoms were invited to participate, and both axillary sweat samples for dog detection and nasopharynx/oropharynx swabs for qPCR were collected. Two dogs, previously trained, detected 97.4% of the samples positive for COVID-19, including a false-negative qPCR-test, and the positive predictive value was 100% and the negative predictive value was 98.2%. Therefore, we can conclude that canine olfactory sensitivity can detect a person infected with COVID-19 through axillary sweat successfully and could be used as an alternative to screen them without invasive testing.

First report of severe acute respiratory syndrome coronavirus 2 detection in two asymptomatic cats in the state of Pernambuco, Northeastern Brazil.

BACKGROUND AND AIM: Despite worldwide case reports, including Brazilian cases, no frequency study on infection of pets by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been conducted to date in Brazil. Accordingly, the present study was aimed to assess dogs and cats belonging to positive owners in Recife, Northeastern Brazil. MATERIALS AND METHODS: This was a longitudinal prospective study on dogs and cats in the city of Recife whose owners were in isolation at home due to a confirmed laboratory diagnosis of SARS-CoV-2 through reverse-transcriptase polymerase chain reaction (RT-qPCR). Oral and rectal swabs from the pets were tested for the presence of SARS-CoV-2-specific RNA by means of RT-qPCR. RESULTS: Among the pets tested, 0/16 dogs and 2/15 cats were positive for SARS-CoV-2. Interestingly, the two positive cats were owned by two unrelated asymptomatic veterinary students, which, therefore, post a warning to veterinarians worldwide. CONCLUSION: The findings herein indicate that cats may act as sentinels for human cases, particularly sharing households with asymptomatic human cases. Although with small sampling and convenient recruiting, the presence of infected cats by SARS-CoV-2 was most likely due to close cat-human contact with positive owners, posting a human-animal health threat when pets share the same bed and interact with owners without protection, particularly during owner self-isolation. Thus, infected owners should follow the same human preventive guidelines with their pets to avoid spreading infection.

A DNA vaccine against yellow fever virus: development and evaluation.

Attenuated yellow fever (YF) virus 17D/17DD vaccines are the only available protection from YF infection, which remains a significant source of morbidity and mortality in the tropical areas of the world. The attenuated YF virus vaccine, which is used worldwide, generates both long-lasting neutralizing antibodies and strong T-cell responses. However, on rare occasions, this vaccine has toxic side effects that can be fatal. This study presents the design of two non-viral DNA-based antigen formulations and the characterization of their expression and immunological properties. The two antigen formulations consist of DNA encoding the full-length envelope protein (p/YFE) or the full-length envelope protein fused to the lysosomal-associated membrane protein signal, LAMP-1 (pL/YFE), aimed at diverting antigen processing/presentation through the major histocompatibility complex II precursor compartments. The immune responses triggered by these formulations were evaluated in H2b and H2d backgrounds, corresponding to the C57Bl/6 and BALB/c mice strains, respectively. Both DNA constructs were able to induce very strong T-cell responses of similar magnitude against almost all epitopes that are also generated by the YF 17DD vaccine. The pL/YFE formulation performed best overall. In addition to the T-cell response, it was also able to stimulate high titers of anti-YF neutralizing antibodies comparable to the levels elicited by the 17DD vaccine. More importantly, the pL/YFE vaccine conferred 100% protection against the YF virus in intracerebrally challenged mice. These results indicate that pL/YFE DNA is an excellent vaccine candidate and should be considered for further developmental studies.

A DNA vaccine against yellow fever virus: development and evaluation

As vacinas 17D / 17DD do vírus atenuado da febre amarela (FA) são a única proteção disponível contra a infecção por FA, que continua sendo uma fonte significativa de morbidade e mortalidade nas áreas tropicais do mundo. A vacina atenuada do vírus da FA, que é usada em todo o mundo, gera tanto anticorpos neutralizantes duradouros quanto respostas fortes de células T. No entanto, em raras ocasiões, esta vacina tem efeitos colaterais tóxicos que podem ser fatais. Este estudo apresenta o projeto de duas formulações antigênicas não-virais baseadas em DNA e a caracterização de suas propriedades imunológicas e de expressão. As duas formulações de antígeno consistem de DNA codificador da proteína de envelope de comprimento total (p / YFE) ou a proteína de envelope de comprimento total fundida ao sinal de proteína de membrana associada ao lisossomo, LAMP-1 (pL / YFE), visando desviar o processamento de antígeno / apresentação através dos compartimentos precursores do complexo principal de histocompatibilidade II. As respostas imunes desencadeadas por estas formulações foram avaliadas em fundos de H2b e H2d, correspondendo às linhagens de camundongos C57Bl / 6 e BALB / c, respectivamente. Ambas as construções de DNA foram capazes de induzir respostas muito fortes de células T de magnitude similar contra quase todos os epítopos que também são gerados pela vacina YF 17DD. A formulação pL / YFE apresentou um melhor desempenho global. Além da resposta das células T, também foi capaz de estimular altos títulos de anticorpos neutralizantes anti-YF comparáveis ​​aos níveis desencadeados pela vacina 17DD. Mais importante, a vacina pL / YFE conferiu 100% de proteção contra o vírus YF em camundongos intracerebralmente desafiados. Estes resultados indicam que o DNA de pL / YFE é um excelente candidato a vacina e deve ser considerado para futuros estudos de desenvolvimento.

Antifungal activity of lectins against yeast of vaginal secretion

Lectins are carbohydrate-binding proteins of non-imune origin. This group of proteins is distributed widely in nature and they have been found in viruses, microorganisms, plants and animals. Lectins of plants have been isolated and characterized according to their chemical, physical-chemical, structural and biological properties. Among their biological activities, we can stress its fungicidal action. It has been previously described the effect of the lectins Dviol, DRL, ConBr and LSL obtained from the seeds of leguminous plants on the growth of yeasts isolated from vaginal secretions. In the present work the experiments were carried out in microtiter plates and the results interpreted by both methods: visual observations and a microplate reader at 530nm. The lectin concentrations varied from 0.5 to 256µg/mL, and the inoculum was established between 65-70% of trammitance. All yeast samples isolated from vaginal secretion were evaluated taxonomically, where were observed macroscopic and microscopic characteristics to each species. The LSL lectin did not demonstrate any antifungal activity to any isolate studied. The other lectins DRL, ConBr and DvioL, showed antifungal potential against yeast isolated from vaginal secretion. These findings offering offer a promising field of investigation to develop new therapeutic strategies against vaginal yeast infections, collaborating to improve women's health.

Heavy metal biosorption by chitin and chitosan isolated from Cunninghamella elegans (IFM 46109)

Chitin and chitosan were extracted from mycelial biomass of Cunninghamella elegans and the performance for copper, lead and iron biosorption in aqueous solution was evaluated. The growth curve of C. elegans was accomplished by determination of biomass, pH, glucose and nitrogen consumption. Chitin and chitosan were extracted by alkali-acid treatment and the yields were 23.8 and 7.8 percent, respectively. For the adsorption analysis, the process of heavy uptake metal sorption was evaluated using polysaccharides solutions (1 percent w/v). The rate of metallic biosorption was dependent upon the concentration and pH of metal solutions, and the best results were observed with pH 4.0. Chitosan showed the highest affinity for copper and chitin for iron adsorption. The results suggest that C. elegans (IFM 46109) is an attractive source of production of chitin and chitosan, with a great potential of heavy metals bioremediation in polluted environments.