Table 1 Study time points

Recruitment

(ANC visit)

–  Full questionnaire administered by a trained healthcare worker to collect information on demographic (eg. age, area of residence), socioeconomic factors (eg. education, occupation), number of previous pregnancies,, bed net ownership and use, adherence to malaria chemoprevention, barriers to administration of IPTp, use of ITNs, risk perception of COVID-19 and Malaria will be administered

–  Clinical examination assessing the general wellbeing and nutritional status of the woman, along with routine measurements (including weight, height, auscultation, blood pressure and temperature). Gestational age will be assessed, when available, by obstretic ultrasound and, if unavailable, by pelvis examination performed by experienced midwives

–  Collection of blood sample that will be analyzed as follows:

oMalaria diagnostic test with microscopy and, when positive, parasite count. Positive samples will be sent to the ISS. Molecular diagnosis will be performed to confirm microscopy results and to discriminate between Plasmodium species. Plasmodium falciparum positive samples will be analyzed for detection of single nucleotide polymorphisms in P. falciparum genes associated with artemisinin and SP resistance. Women diagnosed with malaria will be treated and followed according to Uganda Clinical Guidelines

 o HIV diagnostic test

 o Hemoglobin levels

 o Blood glucose levels

 o Syphilis testing (RPR)

All women found to be HIV + at study entry will be referred for further evaluation and treatment

ANC visits

–  Physical examination

–  Tests to be performed as per study procedure:

–  HB estimation at first contact and at 26 weeks with every pregnant woman:

–  HIV testing: first contact and 36w contact

–  Malaria screening: each ANC visit (maximum 8 times). Malaria diagnostic test with microscopy and, when positive, parasite count. Positive samples will be sent to the ISS. Molecular diagnosis will be performed to confirm microscopy results and to discriminate between Plasmodium species. Plasmodium falciparum Positive samples will be analyzed for detection of single nucleotide polymorphisms in P. falciparum genes associated with artemisinin and SP resistance. Women diagnosed with malaria will be treated and followed according to Uganda Clinical Guidelines

Any spontaneous visits to the hospital related with pregnancy and/or malaria

–  Standardized history

–  Physical exam including temperature, pulse, and blood pressure measurement

–  Patients who are febrile (tympanic temperature > 38.0˚C) or report history of fever in the past 24 h will have blood obtained by finger prick for a thick blood smear. If the thick blood smear is positive, the patient will be diagnosed with malaria. If the thick blood smear is negative, the patient will be managed by study physicians for a non-malarial febrile illness. If the patient is afebrile and does not report a recent fever, a thick blood smear will not be obtained, except when following routine testing schedules

–  In patients with positive microscopy for P. falciparum, the first, pre-therapy blood sample collected as DBS will be sent for molecular diagnosis confirmation and for genetic analysis of antimalarial resistance to ISS. Furthermore, for hospitalized patients, parasitemia will be reassessed at day 3, as per WHO protocol [27]

–  Recruitment of the subpopulation of non-pregnant individuals will be undertaken at this time point

Delivery

–  Delivery information: study staff will document details of the delivery, including date and time, type of delivery, estimated blood loss and any maternal, obstetrical or neonatal complications

–  Fetal outcomes: stillbirths, low birth weight, preterm birth

–  Infant information: Apgar score and birth weight with calibrated scales. At the time of delivery, women will undergo repeat rapid HIV testing based on national guidelines. If women are found to have become HIV-infected during pregnancy, both the mother and their newborn will be referred for care following local prevention of mother-to-child transmission guidelines [26]

–  Analyses of the Placenta Blood by microscopy to detect placental parasitemia