The Hohoe Municipality is one of the twenty-five administrative districts/municipalities in the Volta Region . The Municipality lies within the middle zone of the region and shares boarders on the East with the Republic of Togo, on the Southwest with Kpando Municipality, Northwest with Biakoye District, on the North with Jasikan District, and on the South with Afadzato South District . The Municipality consists of 102 communities with a population of 167,016 people and a population density of 196.0 persons per square kilometres .
This descriptive cross-sectional study recruited 409 pregnant women (6 weeks to 36 weeks of gestation) and 194 postnatal mothers (6 weeks post) of ANC and PNC centres of Hohoe Municipal Hospital for the period March 2017.
We estimated the sample sizes for pregnant women using the regional prevalence of 47%  and 14% for post-partum mothers , using the Cochrane formula . Assuming z-statistic for 95% level of confidence and a 5% margin of error, the appropriate minimum sample size was estimated for the study.
Adjusting for a non-response rate of 5%, a total sample size of 402 was reached for pregnant women and 194 for post-partum mothers.
During data collection, thirty (30) pregnant women were randomly selected each day by balloting without replacement method. The balloting method allowed consented participants to either pick “Yes” or “No” of folded pieces that were placed in a container and thoroughly shaken to ensure randomization. Data was collected from participants who picked “Yes”. This was repeated until the desired sample size was attained. The same sampling procedure was done with regards to the postnatal mothers. Data were collected at PNC clinic and post-delivery wards of the Hospital.
With the aid of trained research assistants, data on socio-demographic characteristics and risk factors associated with anaemia (independent variables) in both pregnant and postpartum mothers were collected using a pre-tested semi-structured questionnaire. Haemoglobin concentration (Hb) were determined by finger-pricked blood test samples of participants using URIT-12 Haemoglobin photometer (URIT Medical Electronics Co., LTD, China). Anaemia was defined as Hb level lower than 11.0 g/dl (Hb < 11.0 g/dl) in pregnant women  and Hb level lower than 10.0 g/dl in postpartum mothers (Hb < 10.0 g/dl) . Capillary blood sampling from the finger was used because it provides a reliable and suitable alternative for sampling blood in the clinical and field settings [22,23,24].
Estimated blood loss data of postpartum mothers were obtained from the maternal delivery records. Blood loss volume of ≤300 ml was considered normal and > 300 ml–500 ml was considered abnormal. Blood loss volume (>500mls) was considered postpartum haemorrhage (PPH) .
Parasitaemia in blood samples were detected using standardized blood film and staining procedures . Three drops of blood were placed on a clean, dust free and dry frosted microscopic slide for thick blood film. Also a drop of blood was placed on the side of the thick film for the preparation of a thin film. A unique Identification number (ID) for each participant and date were written on each slide for easy identification. The slides were air dried and packed into slide boxes and transported to the SPH, UHAS laboratory. The dried slides were stained with 1% Giemsa stain for about 25–30 min. Buffered water (pH = 7) was used to rinse the stained slides. The prepared slides were examined under oil immersion with a light microscope (ocular magnification × 100). The thick film was used for the quantification of the malaria parasites while the thin film was used for identifying the malaria species. Parasite densities were estimated by counting the number of parasites per 200 white blood cells (WBCs) in a thick film by two microscopists. Counts of gametocyte were taken against 500 white blood cells in determining gametocyte density per microliter of blood.
Light microscope was used to read the slides, a sample was considered negative only after 200 high power fields have been read. Parasite counts were converted to parasite per μ1, with the assumption that there is an average of 8000 leucocytes per μ1 of blood. In cases where there was a 50% discrepancy between parasite counts or when there was a discrepancy qualitatively (negative versus positive), a third microscopist read the slide and his reading was final and was used in the analysis of parasite density. All slides were stored in appropriately labelled slide boxes and kept at the laboratory. As part of the quality control monitoring, randomly selected stained slides from each batch of slides were given to an independent microscopist at the Municipal hospital for the determination of the sensitivity and specificity of the readers. Hb readings were quality controlled by trained laboratory scientists from the School of Public Health (SPH) of the University of Health and Allied Sciences (UHAS), research laboratory throughout the study period.
This study was conducted in accordance with accepted principles on ethics in human experimentation and international conference on Harmonization/Good Clinical Practice (ICH/GCP). Ethical approval for the study was obtained from the Ethics Review Committee (ERC) of the University of Health and Allied Sciences (UHAS) with Ethical Approval number UHAS-ERC A.6  17–18. Permission was sought from the Hohoe Municipal Hospital before the commencement of the study. Written informed consent was obtained from participants on standard consent form before they were included in the study.
The data were entered into EpiData version 3.0 and exported to Stata version 14.1 for analysis. Descriptive statistics, frequencies and percentages were used for categorical variables. Normality was determined for continuous variables such Age, Hb, gravidity, parity and family size. Mean ± SD was determined for continuous variables using t-test. Chi-square test was used to determine association between independent variables (socio-demographic characteristics and risk factors) and the dependent variable (anaemia status). A univariate logistic regression was used to determine the strength of the association between the independent and dependent variable. The dependent variable considered in the univariate logistic regression was anaemia status. However, a multivariable logistic regression could not be used because only one variable in the univariate logistic regression showed a statistically significant association with the dependent variable (Anaemia status). Therefore, there was no need to use a multivariable logistic regression. A p-value less than 0.05 was considered statistically significant at 95% Confidence Interval (CI).