Combur 10 test m test strips

The day before sacrifice, all animals were fasted and housed in metabolic cages designed for urine harvest, for approximately 15 h. Then, blood samples were obtained previous to sacrifice: (1) from the retro orbicular plexus for hematology and serum biochemistry, and (2) from the tail vein for coagulation.
Blood samples for hematology were poured into an EDTAK₂ tube. Analyses were conducted with an automatic hematologic analyzer Sysmex XT-1800i™ (LabX, ON, Canada). Also, blood extensions were obtained from each animal and stained with Brilliant Cresyl Blue to perform the reticulocyte count. Blood samples for serum biochemistry were poured into a separator gel tube and centrifuged (1500× g, 10 min, 20 °C). The serum was obtained and analyzed with a biochemical autoanalyzer Cobas c-311(Roche, Basilea, Switzerland). Blood samples for coagulation were poured into a sodium citrate (0.109 M) tube. After centrifugation, serum was obtained and analyzed with a coagulometer STart4 (GMI Inc., Ramsey, MN, USA). The urine was collected and analyzed using Combur 10 Test^® M test strips with a Cobas u 411^® analyzer (Roche, Basilea, Switzerland).

The participants collected at least 35 ml of their first-morning urine. At the workstation, the samples underwent a chemical analysis using Combur10 Test^® M test strips (Roche, Basel, Switzerland) to check and record any alterations in pH value, and detect the presence of leucocytes, protein, hemoglobin, and/or nitrite. This step was conducted by two experienced researchers simultaneously. For that, a test strip was immersed into every urine sample for about 1 s and then, after 1 min, the color presented on each pad of the strip was compared to the reference scale. The interpretation was conducted according to the manufacturer’s instructions, and the Guidelines from the Brazilian Society for Clinical Pathology and Laboratory Medicine (SBPC/ML; Andriolo et al., 2017 ). After that, two drops of each urine sample were used for the POC-CCA test. The remaining volume was filtered in a cone folded Whatman^® qualitative filter paper, Grade 3:6 μm (diameter 185 mm), previously labelled with the participant ID. After the filtration, the paper filters were left exposed on a sterilized bench to completely dry. They were then individually stored in hermetically sealed plastic bags, together with desiccant silica gel, and then transported at room temperature from the field to the laboratory, where the DNA extraction and NAATs tests were conducted.