High levels of pro-inflammatory SARS-CoV-2-specific biomarkers revealed by in vitro whole blood cytokine release assay (CRA) in recovered and long-COVID-19 patients.

BACKGROUND: Cytokines induced by SARS-CoV-2 infection play a crucial role in the pathophysiology of COVID-19 and hyperinflammatory responses have been associated with poor clinical outcomes, with progression to severe conditions or long-term subacute complications named as long-COVID-19. METHODS: In this cross-sectional study, we aimed to evaluate a set of antigen-specific inflammatory cytokines in blood from recovered COVID-19 individuals or who suffered a post-acute phase of SARS-CoV-2 infection compared to healthy individuals with no history of COVID-19 exposition or infection. Interferon-gamma (IFN-γ), IFN-γ-induced protein 10 (IP-10), tumor necrosis factor (TNF), IL-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, and IL-17A were quantified by multiplex cytometric bead assay and enzyme-linked immunosorbent assay after stimulation of whole blood with recombinant Spike protein from SARS-CoV-2. Additionally, all participants have evaluated for anti-(S) protein-specific IgG antibodies. Clinical specimens were collected within two months of COVID-19 diagnosis. RESULTS: A total of 47 individuals were enrolled in the study, a median age of 43 years (IQR = 14.5), grouped into healthy individuals with no history of infection or exposure to SARS-CoV-2 (unexposed group; N = 21); and patients from the Health Complex of the Rio de Janeiro State University (UERJ), Brazil, who were SARS-CoV-2 positive by RT-PCR (COVID-19 group)-categorized as recovered COVID-19 (N = 11) or long-COVID-19 (N = 15). All COVID-19 patients presented at least one signal or symptom during the first two weeks of infection. Six patients were hospitalized and required invasive mechanical ventilation. Our results showed that COVID-19 patients had significantly higher levels of IFN-γ, TNF, IL-1ß, IL-2, IL-6, IL-8, and IP-10 than the unexposed group. The long-COVID-19 group has presented significantly high levels of IL-1ß and IL-6 compared to unexposed individuals, but not from recovered COVID-19. A principal-component analysis demonstrated 84.3% of the total variance of inflammatory-SARS-CoV-2 response in the first two components, and it was possible to stratify IL-6, TNF, IL-1ß, IL-10, and IL-2 as the top-five cytokines which are candidates to discriminate COVID-19 group (including long-COVID-19 subgroup) and healthy unexposed individuals. CONCLUSION: We revealed important S protein-specific differential biomarkers in individuals affected by COVID-19, bringing new insights into the inflammatory status or SARS-CoV-2 exposition determination.

Two for the price of one: Co-infection with Rickettsia bellii and spotted fever group Rickettsia in Amblyomma (Acari: Ixodidae) ticks recovered from wild birds in Brazil.

The bacterium Rickettsia bellii has been detected in 25 species of ticks in the American continents, but its pathogenic potential is considered as undetermined. A possible role for this species in the phenomenon of transovarial exclusion of pathogenic members of the spotted fever group (SFG) of Rickettsia has been suggested and co-infections with pathogenic species have been reported infrequently in both North and South America. Traditional methods for the molecular detection of rickettsial agents in ticks focus largely on the identification of sequences found in SFG Rickettsia, an approach that may overlook the presence of co-infections with R. bellii. Two novel, species-specific polymerase chain reaction (PCR) assays, targeting the genes encoding the surface cell antigen (Sca), autotransporter proteins sca9 and sca14, were developed and validated for the detection of R. bellii using 150 Amblyomma ticks collected from wild birds in Brazil. Co-infection of R. bellii infected ticks was evaluated using a novel PCR assay targeting the ompA sequence characteristic of SFG Rickettsia. Preliminary species-level identification was achieved by restriction fragment length polymorphism (RFLP) analysis and subsequently confirmed by sequencing of amplicons. Nine out of seventy-three Amblyomma longirostre and one of two Amblyomma calcaratum ticks were shown to be co-infected with R. bellii and Rickettsia amblyommatis, while two out of sixty-seven Amblyomma sp. haplotype Nazaré ticks were recorded as co-infected with R. bellii and the Rickettsia parkeri-like bacterium, strain ApPR. Interestingly, our data represent the first records of R. bellii in association with A. calcaratum and Amblyomma sp. haplotype Nazaré. The novel PCR-RFLP systems reported herein, provide an alternative, rapid and cost-efficient (relative to strategies based on sequencing or real-time PCR), approach to evaluate rickettsial co-infection of ticks, a potentially significant phenomenon that has most likely been underestimated to date.

Contaminação fúngica no leite humano e em sítios anatômicos de lactantes e lactentes / Fungal contamination in human milk and in the anatomic sites of breastfeeding mothers and infants

As infecções bacterianas ou fúngicas causam quadro clínico de mastite, que motiva desmame precoce. Os micro-organismos patogênicos, como leveduras do gênero Candida, quando em número elevado no intestino, podem causar disbiose. Nesta pesquisa, foram realizadas a detecção e a identificação de microbiota fúngica nas amostras de leite humano e de sítios anatômicos de mulheres e crianças atendidas pelo Banco de Leite Humano do Instituto Fernandes Figueira. A virulência dos isolados de levedura foi determinada pelos testes de atividade proteolítica. De 64 amostras analisadas, 81% foram positivas para fungos, com maior prevalência de Candida albicans (73%), seguida do complexo C. parapsilosis (15,4%). Perfis semelhantes aos verificados no total de amostras foram encontrados nas amostras de leite, nas mamas e na cavidade oral, sugerindo-se a ocorrência de associação entre a infecção cutânea da mãe e do lactente com o leite ingerido. O perfil associado à virulência dos isolados de Candida foi determinado pelo teste de produção de proteases, e 100% das amostras mostraram resultados fortemente positivos, indicando alto grau de infecciosidade. A alta prevalência de C. albicans nas amostras coletadas de mamas, no leite e na cavidade oral, é importante fator de risco à saúde de lactentes.

Cryptococcus neoformans isolados de Periplaneta americana recolhidas de ambientes públicos e sua relevância para saúde humana e animal / Cryptococcus neoformans isolated from Periplaneta americana collected from public environments and its relevance to human and animal health

O objetivo deste trabalho foi verificar a presença de Cryptococcus neoformans em baratas (Periplaneta americana) recolhidas de ambientes públicos. A motivação para esta pesquisa deve-se ao fato de esta espécie de barata sersinantrópica e veiculadora de vários agentes patogênicos para o homem e animais, o que é relevante para a saúde pública. Os insetos foram recolhidos vivos, mediante emprego de armadilhas, e também mortos, totalizando 150 amostras. A pesquisa da presença do agente nos insetos foi feita no Laboratório de Leveduras Patogênicas e Ambientais, Departamento de Microbiologia e Imunologia Veterinária do Instituto de Veterinária da UFRRJ, por processamento do exoesqueleto e de seu conteúdo interno, com semeadura em meio de cultivo contendo dopamina. As confirmações de isolamento foram realizadas por meio de testes de macromorfologia, micromorfologia, auxanograma e provas complementares. Não se obteve o isolamento de Cryptococcus neoformans do exoesqueleto; os cinco obtidos (3,3por cento) foram do conteúdo interno dos insetos que, após sorogrupagem em meio CGB,revelaram-se pertencentes ao sorogrupo AD. As baratas da espécie Periplaneta americana são, portanto, consideradas um potencial vetor para a levedura estudada.

The objective of this paper was to check the presence of Cryptococcus neoformans in cockroaches (Periplaneta Americana) collected in public places. The research’s motivation was based on the fact that this species of cockroach are synanthropic and vehicle of several pathogenic agents for men and animals, a relevant fact for public health. A total of 150 samples were collected and the research for the presence of the agents in the insects was conducted in the Laboratory of Pathogenic and Environmental Yeasts, Department of Microbiology and Immunology Veterinary,UFRRJ, by processing of the exoskeleton and the internal content and sowing in cultivation medium containing dopamine. The confirmation of isolation was made by the macromorphology and micromorphology tests, performance ofauxanographic method, and complementary proofs. Isolation of Cryptococcus neoformans was not obtained from the exoskeleton. Five isolates (3.3%) were obtained from the internal content of the insects. The serogrouping in CGB medium showed that they were from the AD serum group. We consider that the cockroachesof the species Periplaneta americana are a potential vector for the studied yeast.