Multiplexed flow cytometric approach for detection of anti-SARS-CoV-2 IgG, IgM and IgA using beads covalently coupled to the nucleocapsid protein.

Flow cytometry has emerged as a promising technique for detection of SARS-CoV-2 antibodies. In this study, we developed an innovative strategy for simultaneous detection of immunoglobulin G (IgG), IgM and IgA. The SARS-CoV-2 nucleocapsid protein was covalently bound to functional beads surface applying sulpho-SMCC chemistry. BUV395 anti-IgG, BB515 anti-IgM, biotinylated anti-IgA1/IgA2 and BV421 streptavidin were used as fluorophore conjugated secondary antibodies. Serum and antibodies reaction conditions were optimized for each antibody isotype detection and a multiplexed detection assay was developed. This new cell-free assay efficiently discriminate COVID-19 negative and positive samples. The simultaneous detection of IgG, IgM and IgA showed a sensitivity of 88·5-96·2% and specificity of 100%. This novel strategy opens a new avenue for flow cytometry-based diagnosis.

Urinary parameters of Trichechus inunguis (Mammalia, Sirenia): reference values for the Amazonian Manatee.

The Amazonian manatee, Trichechus inunguis (Natterer 1883) is endemic to the Amazon Basin and is currently considered a vulnerable species. In order to establish normality ranges of urinary parameters to help monitor the health of this species in captivity, chemical urinalyses were performed on twelve males and nine females of various age groups. Urine was collected once a month for twelve months in the tanks just after being drained, by placing stainless steel containers under the genital slit of females and applying abdominal massages to males in order to stimulate urination. Quantitative data of glucose, urea, creatinine, uric acid and amylase levels were obtained using colorimetric spectrophotometry. Dip strips were also useful for routine analyses, despite only providing qualitative results. Normal range to glucose levels, regardless of sex or age class, was 3.0 to 3.6 mgxdL-1, coinciding with qualitative values of glucose measured by dip strips. Statistical differences observed in some parameter levels suggest that some urine parameters analysed must take into consideration the sex and the age class of the animal studied, being these differences less remarkable in creatinine and amylase levels. To this last one, statistical difference was detected only in the calve's urine (7.0 to 11.5 mgxdL-1) compared to other age classes samples (4.1 to 5.3 mgxdL-1). The results presented here may be used as comparative data in future research on urinalysis in related species.

Urinary parameters of Trichechus inunguis (Mammalia, Sirenia): reference values for the Amazonian Manatee / Parâmetros urinários de Trichechus Inunguis (Mammalia, Sirenia): valores-referência para o peixe-boi da Amazônia

The Amazonian manatee, Trichechus inunguis (Natterer 1883) is endemic to the Amazon Basin and is currently considered a vulnerable species. In order to establish normality ranges of urinary parameters to help monitor the health of this species in captivity, chemical urinalyses were performed on twelve males and nine females of various age groups. Urine was collected once a month for twelve months in the tanks just after being drained, by placing stainless steel containers under the genital slit of females and applying abdominal massages to males in order to stimulate urination. Quantitative data of glucose, urea, creatinine, uric acid and amylase levels were obtained using colorimetric spectrophotometry. Dip strips were also useful for routine analyses, despite only providing qualitative results. Normal range to glucose levels, regardless of sex or age class, was 3.0 to 3.6 mg.dL-1, coinciding with qualitative values of glucose measured by dip strips. Statistical differences observed in some parameter levels suggest that some urine parameters analysed must take into consideration the sex and the age class of the animal studied, being these differences less remarkable in creatinine and amylase levels. To this last one, statistical difference was detected only in the calve's urine (7.0 to 11.5 mg.dL-1) compared to other age classes samples (4.1 to 5.3 mg.dL-1). The results presented here may be used as comparative data in future research on urinalysis in related species.

O peixe-boi da Amazônia, Trichechus inunguis (Natterer 1883) é endêmico da Bacia Amazônica e é considerado atualmente como espécie vulnerável. Com o objetivo de estabelecer valores de normalidade para os parâmetros urinários com vistas a ajudar no monitoramento da saúde desta espécie em cativeiro, foi realizada urinálise química em doze machos e nove fêmeas de várias classes etárias. A urina era coletada uma vez por mês, durante doze meses, nos tanques, logo após esvaziamento, colocando-se coletores de aço inoxidável sob o orifício genital das fêmeas e aplicando massagem abdominal nos machos para estimular a micção. Dados quantitativos dos níveis de glicose, ureia, creatinina, ácido úrico e amilase foram obtidos por meio de espectrofotometria colorimétrica. O uso de tiras reativas também foi útil para análises de rotina, apesar de somente fornecerem resultados qualitativos. A faixa de normalidade para os níveis de glicose, independente de classe etária e sexo foi de 3,0 a 3,6 mg.dL-1, coincidindo com os valores qualitativos identificados pelas tiras reativas. Diferenças estatísticas observadas nos níveis de alguns parâmetros analisados sugerem que estes devem levar em consideração o sexo e a classe etária do animal estudado. Estas diferenças foram menos marcantes nos níveis de creatinina e de amilase, sendo que para esta última foi detectada diferença estatística apenas nas urinas de filhotes lactentes (7,0 a 11,5 mg.dL-1), em comparação às amostras das demais classes etárias (4,1 a 5,3 mg.dL-1). Os resultados aqui apresentados podem ser usados como dados comparativos em futuras pesquisas sobre urinálise em espécies relacionadas.