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PLOS One logoLink to PLOS One
. 2023 May 30;18(5):e0285977. doi: 10.1371/journal.pone.0285977

The prevalence of urogenital and intestinal schistosomiasis among school age children (6–13 years) in the Okavango Delta in Botswana

Nthabiseng A Phaladze 1,*, Lebotse Molefi 2, Olekae T Thakadu 3, Onalenna Tsima 1, Barbara N Ngwenya 3, Tuduetso L Molefi 4, Wananani B Tshiamo 1
Editor: Clement Ameh Yaro5
PMCID: PMC10228809  PMID: 37253026

Abstract

This study sought to investigate prevalence of urogenital and intestinal schistosomiasis among school age children 6–13 years in selected communities in the Okavango Delta. The termination of the Botswana national schistosomiasis control program in 1993 contributed to its neglect. An outbreak of schistosomiasis in 2017 at one of the primary schools in the northeastern part of the country resulted in 42 positive cases, indicating that the disease exists. A total of 1,611 school age children 6–13 years were randomly selected from school registers in 10 primary schools; from which 1603 urine and 1404 stool samples were collected. Macroscopic examination of urine and stool for color, odor, blood; viscosity, consistency, and the presence of worms. Urine filtration and centrifugation methods were used to increase sensitivity of detecting parasite ova. Kato-Katz and Formalin-Ether were used for the examination of stool samples. Data were analyzed using SPSS version 25. Results were expressed as odds ratio (OR) with their 95% CI and statistical significance set at p < 0.05. A total of (n = 1611) school age children 6–13 years participated in the study, mean age 9.7years (SD 2.06), females (54%) and males (46%). Results indicated an overall prevalence of SS. hematobium and S.mansoni at 8.7% and 0.64% respectively. Intensity of SS. hematobium was generally light (97.6%) and heavy intensity (2.4%). Results also revealed a knowledge deficit, about 58% of children had never heard of bilharzia even though they lived in communities where the disease was previously endemic. Learners who had a family member who previously suffered from schistosomiasis had higher knowledge than those who did not. Interestingly, these learners were likely to engage in risky behaviors compared to those with lower knowledge of the disease. An integrated approach that emphasizes health education, mass drug administration, water, sanitation, and hygiene infrastructure should be prioritized for prevention and control of schistosomiasis.

1. Introduction

Schistosomiasis is considered the second most neglected tropical disease (NTD) after hookworm. It mainly affects developing countries where water resources and poor sanitation allow development and infection of snails respectively [1]. Schistosomiasis affects a large proportion of children under 14 years of age, including at least 25 million preschool aged going children [2]. Schistosomiasis is a parasitic disease caused by blood vessel-dwelling flukes of the genus schistosoma. There are several species in the genus but primarily S. hematobium which causes urinary schistosomiasis, S. mansoni and S. japonicum which both cause intestinal schistosomiasis in humans [3]. are the most prevalent. The invading parasites in Botswana are S. mansoni and S. haematobium, with prevalence rates fluctuating depending on rainfall. Prevalence of schistosomiasis in Botswana has been as high as 80% or as low as 20% in defined geographical areas [4].

The effects of schistosomiasis among children are diverse and alarming, such as urethral and bladder fibrosis, hydronephrosis, hepatosplenomegaly, and can progress into bladder cancer and colorectal cancer as a possible late-stage complication [5, 6]. found that schistosomiasis has profound negative effects on child development, outcome of pregnancy, and agricultural productivity, thus a key reason why the most inhabitants of sub-Saharan Africa continue to live in poverty. Schistosomiasis causes inflammation of the mucosa, hyperplasia of cells and gut ulceration [7], which in turn decreases food intake and nutrient utilization within the host body [810], all of which contribute to childhood stunting [11]. The disease can lead to chronic ill health and is considered a major public health concern mostly in rural dwellers of tropical and sub-tropical regions of the world [12].

Human contact with snail habitats is crucial to the establishment of transmission and children are more frequently and more severely infected than adults. Swimming is a water contact activity that contributes most transmission. Children swim often, especially during warmer months and invariably urinate while doing so, thus introducing continual supply directly into the water [13]. However, a clear understanding of the snail species, their local distribution and infection status is therefore essential for effective control of schistosomiasis [14].

Previous KAP studies have shown differential knowledge levels among communities. A systematic literature review on schistosomiasis revealed that awareness and knowledge was low mainly among rural communities [15]. Other studies conducted in Ethiopia, Ghana, Côte d’Ivoire, Tanzania, and Mozambique also showed low levels of knowledge of the diseases [1618] while other studies conducted in Zimbabwe and Kenya showed higher level of knowledge [18]. The studies concluded that low levels of knowledge of the disease makes communities vulnerable and at risk of infection. Others [19, 20] explored the effect of gender on risk factors, more so that gender roles may dictate the frequency of water contacts among males and females, thereby pointing to exposure to risky behaviors. A study in Uganda found no differences among boys and girls in the frequency of water contact activities, while in adults, women had more water activities compared to men [20]. A meta-analysis revealed that males were more vulnerable to reinfection than females [19]. Other demographic characteristics showing strong association with reinfection included age [21].

In Botswana, the highest prevalence of S. mansoni infection (just over 80%) was recorded in 1983, 6 years after the exceptionally high in-flows of 1977. This wet period was followed by the dry conditions of the 1980s, including several dry spells where the Thamalakane River did not receive water for several consecutive months. This decline in flow was accompanied by a decline in the prevalence of S. mansoni infection, with low values recorded in 1989 (8.4%) and 1991 (6.7%) that match the declines in flow seen 6 years prior to each of these dates [4]. Comparison of flows with the prevalence of schistosome infection in young school children indicates that there is an inverse relationship between the rise of flow and fall of schistosome transmission, Appleton and colleagues further noted that there was little evidence of the active transmission of S. hematobium in the delta over the last few decades [4]. None of the communities in Ngamiland district mapped by Doumenge in 1987 had prevalence of S. haematobium infection exceeding 6.8% [22], although a prevalence of 32% at Pandamatenga was recorded [23]. This dominance of S. mansoni over S. haematobium is not confined to the Okavango Delta.

In Botswana, the termination of the National Schistosomiasis Control Program in 1993 contributed to its neglect. However, an outbreak of schistosomiasis in 2017 at one of the primary schools in the northeastern part of the country, resulted in 42 positive cases, an indication that the disease exists. Therefore, this study sought to determine the prevalence of urogenital and intestinal schistosomiasis among school age children 6 to 13 years in selected communities in the Okavango Delta.

2. Methods

2.1 Study design

We used the previous minimum estimates of the prevalence found by Appleton and colleagues [4] to estimate the minimum required sample size for a cross-sectional study design. With the prevalence of 20%, probability of type 1 error of 0.05 and the margin of error of 0.05, the estimated minimum sample size was 246. Assuming a non-response of 20%, the minimum sample size required is 308. However, a descriptive cross-sectional survey was administered to a total of 1,611 school age children 6–13 years who were randomly selected from the school registers in 10 primary schools from a population.

2.2 Study area and population

The study villages Kauxwi, Shakawe, Etsha 6 and Maun were purposively sampled based on their location across the three sections of the Okavango Delta: upper, middle and the distal part.

Kauxwi is situated on the north-eastern part of the panhandle and Shakawe on the south-western side. The two villages are about 10 km apart across the Okavango River. Etsha 6 is situated in the middle section, while Maun, the major urban centre, is in the south-eastern side, the distal section of the Okavango Delta, straddling the Thamalakane River. The three different sections of the Okavango Delta, on which the study sites are located, present varied hydrological conditions and flooding regimes that may pose a risk to bilharzia.

Secondly, schools within the four study sites were also purposively sampled based on their location to the river. Kauxwi Primary School was the only primary school in the village, while Shakawe village had two primary schools, Shakawe and Kathiana which were all selected because of their proximity to the river. Etsha 6 Primary School was selected as the only primary school in Etsha 6. In Maun, six primary schools were selected for being the closest to the Thamalakane River. These were Mathiba, Botswelelo, Thamalakane, Moremi, Letsholathebe and Thito Primary.

Lastly, respondents were sampled using stratified random sampling procedure based on grade or standard level to ensure representativeness [24]. School registers were sought from the school management, and where a grade had more than one class, the names were combined into one list to make one sampling frame. From each grade level, 40% of the children were randomly sampled using computer generated random numbers to take part in the school survey. Sample sizes per school are indicated in Table 1.

Table 1. Total enrolment of school age children in the 4 study sites and actual sample selected.

Village Primary Schools Total Enrolment Total Sample Actual Sample
Maun Letsholathebe 678 272 206
Moremi 607 242 112
Thito 211 85 55
Botswelelo 438 175 115
Thamalakane 363 142 110
Mathiba 908 363 275
Etsha 6 Etsha 6 635 254 185
Shakawe Kathiana 702 283 111
Shakawe 721 291 262
Kauxwi Kauxwi 465 187 180
Total 5728 2294 1611

2.3 Ethical considerations

Ethical approval was obtained from the University of Botswana Institutional Review Board and the Health Research Development Committee of the Ministry of Health and Wellness (HPDME: 13/18/1). Permission was also sought from the Botswana Ministry of Basic Education, Ngamiland Regional Education Office. We obtained written consent from parents and legal guardians and oral assent from children. The purpose of the study and the interview process were described, and the interviewers ensured that the children were comfortable. The assent form was read to the children, and they were requested to complete the form if they agreed to participate in the study, which was completely voluntary. They were assured of confidentiality and that they had the right not to answer any or some of the questions that made them feel uncomfortable and that they had the right to withdraw from the study at any time without penalty. Data were analyzed anonymously. Children who were found positive for S. mansoni and S. hematobium were treated according to the WHO clinical guidelines and Ministry of Health and Wellness.

2.4 Data collection

2.4.1 Parasitology

A total of 1603 urine and 1404 stool samples were collected from school age children in the selected schools to determine the prevalence and intensity of schistosomiasis. Macroscopic examination of urine for color, odor and blood was conducted for all urine samples. 10ml of urine was processed by filtration and examined for parasite ova at x40 magnification for schistosoma ova. To increase the sensitivity of detecting parasite ova, urine filtration and centrifugation methods were used to analyze urine samples concurrently [2527]. Positive diagnosis of S. haematobium was based on the detection of one terminal spined schistosoma ovum or more.

Kato-Katz and Formalin-Ether methods were used for the examination of stool samples. The technique was used to detect the presence of S. mansoni and other parasite ova in single stool specimens. All Stool samples were examined macroscopically for viscosity, color, odor, consistency, and the presence of worms. Kato-Katz and Formalin-Ether methods were run concurrently on each stool sample collected. The slides were examined under a microscope at x40 objective lens by experienced laboratory technicians and a medical laboratory scientist who counted the number of S. mansoni eggs and STH in the stool. Infection intensity was determined by multiplying the number of ova detected on Kato-katz by 24 to determine the number of eggs per gram(epg) of stool [2830].

2.5 Survey questionnaire

Primary data were collected using a structured survey questionnaire, capturing the socio-demographics of the participant, household water and sanitation facilities, family livelihood, hobbies and recreational activities, information sources, knowledge, and practices regarding bilharzia. The survey instruments were reviewed for face and content validity by a panel of experts within the research team. All questionnaires were translated into Setswana and back translated into English.

2.5.1 Data management and statistical analysis

Data quality assurance was ensured through training of research assistants on basic interviewing skills involving children, data capturing, coding, and processing of parasitology samples. Researchers did random spot-checks to review completed questionnaires. Data was entered into a password protected computer.

Quantitative data were analyzed using SPSS version 25. Means and standard deviations were used to summarize continuous variables while frequencies and proportions were used for categorical data such as knowledge, water, sanitation, and hygiene (WASH) facilities, recreational and parasitology results for the overall sample. To test for differences between categorical variables, we used the Fisher’s Exact test. For continuous ones, we used t-tests and one-way between-group ANOVA to test for significant differences among groups. Logistic regression was conducted to examine the effect of a set of selected predictor variables on likelihood of swimming in stagnant water, being a risky behavior. Assumptions for t-test, ANOVA and logistic regression were tested and found tenable. Prevalence of infection was estimated by dividing the number of individuals who tested positive with the total number of children examined. The results were expressed as odds ratio (OR) with their 95% CI and statistical significance set at p < 0.05.

3. Results

3.1. A total of (n = 1611) school age children 6 to 13 years participated in the study, mean age 9.73 years (SD 2.06), females (54%) and males (46%).

3.2 Parasitology results

The results indicate that Shakawe and Kauxwi Primary Schools had each 34 (12% and 21%) cases of S. haematobium while Kathiana and Estha 6 had 9 and 6 (3%) cases respectively. Cases of S. mansoni were very few ranging between 0.4–1% as indicated in Table 2. Although other soil transmitted helminths were not the focus of this study; Ascaris was highest (n = 20) in Shakawe Primary School than in schools.

Table 2. Parasitology results in the upper panhandle and middle sections of the Okavango Delta.

School Urine (N) SS. hematobium Stool (N) S. mansoni
Shakawe 276 34 260 1
Kathiana 286 9 272 1
Kauwxi 169 34 150 2
Etsha 6 178 6 165 1
909 83 847 5

Parasitology results from Maun primary schools reported few cases of S. haemotobium and S. mansoni and other soil transmitted helminths (see Table 3). Although other soil transmitted helminths were not the focus of this study; Ascaris and Taenia were identified, and about 50% (n = 14) was in Mathiba Primary School.

Table 3. Parasitology results of primary schools in Maun the distal part of the Okavango Delta.

School Urine (N) SS. hematobium Stool (N) S. mansoni
Thamalakane 43 0 43 0
Mathiba 259 0 151 0
Botswelelo 146 2 146 0
Thito 45 0 45 0
Moremi 108 0 87 0
Letsholathebe 93 1 85 2
694 3 557 2

3.3 Prevalence and Intensity of schistosomiasis

The overall prevalence of S. haematobium was 8.7%, and for S. mansoni 0.64% respectively. Other soil transmitted helminthes, Ascaris10.9% and hookworm 1.06%. Intensity for S. haematobium was generally light at 97.6% (n = 40), and heavy intensity 2.4% (n = 1) (see Table 4).

Table 4. Prevalence and Intensity of S. hematobium.

S. Hematobium Infection N (%)
Not infected 430 (91.3)
Infected 41 (8.7)
Light intensity Intensity 40 (97.6)
Heavy intensity 1 (2.4)

The prevalence and intensity of S.mansoni was very low at 0.64% (see Table 5) below, this was also reflected in the parasitology results (Tables 3 and 4).

Table 5. Prevalence and intensity of S. mansoni.

S. mansoni Infection N (%)
Not infected 469 (99.4)
Infected 3 (0.64)
No intensity Intensity 2 (66.7)
Light intensity 1 (33.3)

3.4 Knowledge of schistosomiasis

An independent-samples t-test revealed significant differences in age amongst females and males, with mean age for males (M = 9.95, SD = 2.13) a little bit higher than that of females (Table 6). Most learners 58% (n = 917) had not heard of bilharzia. Although more girls (n = 362, 53.7%) were aware of bilharzia than boys (n = 312, 47%), statistically there was no significant association between gender and awareness of bilharzia (χ2 (1, n = 1590) = .15, p = 0.70, phi = 0.01) An independent-samples t-test conducted to compare knowledge scores for i) bilharzia risk activities, ii) signs and symptoms and iii) overall combined knowledge scores by gender revealed no significance difference in scores for males and females (Table 6). The overall knowledge mean score is 4.71, SD = 2.68, signifying low knowledge of schistosomiasis.

Table 6. Association between selected variable by gender.

Variable N Females (n%) Males (n%) F P Effect
Age 1607 9.59 (1.99) 9.89 (2.13) 2.95 0.003
Awareness of Schistosomiasis 1590 53.7 (362) 46.3 (312) 0.15 0.70 0.01
Knowledge of Bilharzia risk factors 772 2.86 (1.6) 2.81 (1.46) -0.44 0.66
Knowledge of bilharzia Signs and symptoms 649 1.99 (1.50) 1.93 (1.53) -0.44 0.66
Knowledge composite 648 4.76 (2.68) 4.63 (2.69) -0.62 0.54

Superscript1 means p-value for Fishers’ exact test has been reported. Superscript means that the mean, standard deviation, and p-value for t-test have been reported.

Table 6 shows that according to the Fisher’s Exact test, only age was different among the gender groups, p = 0.00. One-way between-groups ANOVA was conducted to examine the effect of grade on knowledge scores of bilharzia, measured on total scores each pupil got correct on bilharzia risk activities, signs and symptoms. Learners were in their pre-existing grade level, being Standard 1, 2, 3, 4, 5, and 6. There were significant statistical differences among the six grade level groups: F (5, 637) = 16.92, p = 0 .00, with a large effect size (eta squared) of 0.11. Post hoc comparisons using Tukey HSD revealed significant differences between Grade 1 and all other Grades; Grade 2 significantly differed with three Grades (1, 3, and 6); Grade 3 with Grades 1 and 2, Grade 4 significantly differed with Grade 1 only; Grade 5 also differed with Grade 1 only while Grade 6 differed significantly with Grades 1 and 2. Assessment of mean scores indicates that Grade 1 scored lowest among all grades (M = 2.24; SD = 2.87) (Table 7). Furthermore, Table 3 shows that the mean scores increased from Grade 1 (being the lowest) to Grade 3, then decreased and almost levelled between Grade 4 and 5 and ultimately increased for Grade 6, which has the highest mean scores.

Table 7. Learners’ knowledge mean scores by grade level.

95% CI
Grade N M SD SE LB UB
1 63 2.24 2.87 0.36 1.52 2.96
2 72 3.92 3.36 0.40 3.13 4.71
3 109 5.07 2.44 0.23 4.61 5.54
4 117 4.98 2.49 0.23 4.53 5.44
5 126 4.90 2.48 0.22 4.47 5.34
6 156 5.46 2.06 0.17 5.13 5.78
Total 643 4.71 2.69 0.11 4.50 4.92

An independence-sample t-test was conducted to compare bilharzia knowledge scores for learners who had had a family member who suffered bilharzia and those who did not. There was a significance difference in mean knowledge scores for those who have had a family member who previously suffered from bilharzia (M = 5.98, SD = 1.90) and those who never had a family member infected with bilharzia (M = 4.56, SD = 2.78; t (80.76) = 4.94, p = 0.00). The effect size was very small (eta squared = 0.04), implying that only 4% of the variance in knowledge scores were explained by whether one had a family member infected with bilharzia or not. Mean knowledge scores for learners who had a family member previously infected with bilharzia was higher than those who did not. The mean difference was 1.42, 95% CI [0.85, 1.99].

Logistic regression was conducted to examine the effect of a set of selected predictor variables on the likelihood of swimming in stagnant water, being a risky behavior. The explanatory variables were grade level, gender, knowledge of bilharzia and whether one had had a family member who suffered from bilharzia while the response variable was whether one has swum in stagnant water or not. The overall model was statistically significant, χ2 (8, N = 626) = 36.01, p < .001. The model was able to correctly predict 72.7% of the cases and this was a small improvement from the null model which only classified 71.9% cases correctly. Knowledge of the disease and previous cases of infection within the family made a unique statistically significant contribution to the model, while grade level and gender did not (Table 8).

Table 8. Logistic regression predicting likelihood of engaging in a risky behavior.

95% CI for Odds Ratio
B SE Wald Df P Odds Ratio Lower Upper
Grade 3.80 5 0.58
Grade 1 0.04 0.42 0.01 1 0.92 1.04 0.46 2.38
Grade 2 -0.49 0.41 1.48 1 0.22 0.61 0.28 1.35
Grade 3 0.01 0.39 0.00 1 0.98 1.01 0.47 2.16
Grade 4 -0.28 0.39 0.50 1 0.48 0.76 0.35 1.63
Grade 5 -0.14 0.38 0.13 1 0.72 0.87 0.42 1.83
Gender 0.23 0.19 1.51 1 0.22 1.26 0.87 1.80
Knowledge of disease 0.16 0.04 19.70 1 0.00 1.19 1.10 1.29
Family member infected -0.76 0.31 5.94 1 0.02 0.47 0.26 0.86
Constant -1.07 0.47 5.26 1 0.02 0.35

The results show the odds ratio of 1.19 for knowledge of disease (bilharzia), indicating that for every additional unit of knowledge of bilharzia, the odds were 1.19 times higher that respondents would report swimming in stagnant water, controlling for other factors. This shows that among the learners, the higher the knowledge of bilharzia, the higher the likelihood of engaging in risky behaviors, such as swimming in stagnant water. For learners whose family members had suffered from bilharzia, the odds of swimming in stagnant water were 0.47 lower than those who did not have a family member infected with bilharzia. This implies that respondents who had family members infected with bilharzia were less likely to report swimming in stagnant water, thereby engaging in risky behavior.

3.5 Availability of sanitation and water facilities

Regarding availability of water and sanitation facilities, 55% used pit latrines, 23% the bush and 21.3% flush toilets. Those who used pit latrines (n = 879) and those who used the bush were at higher risk of contracting infection n = 366 (22.7%), OR = 0.7 (CI; 0.0–0.8), and [343 (21.3%), OR = 0.5(CI;0.0–0.9)]. The findings also indicate that about 49% accessed water from the river, while 26.9% and 1.4% accessed it from reserve tanks (Jojo) and boreholes respectively. Most (80.6%) reported that swimming in infected water, drinking dirty water (75%), walking barefooted in dirty water (75%) and washing clothes in dirty water (53%) could transmit infection.

In addition, 43.1% reported that they swim; of these 70.3% swam in the river or stream and that they swam over weekends and school holidays. However, only a few (6.2%, n = 77) reported previous infection, while (6.7%) had a family member diagnosed with schistosomiasis in the past.

4. Discussion

Since the termination of the national program in 1993, schistosomiasis has received little attention. The termination of the program has over the years hampered the goals set by the World Health Assembly Resolution [31] to be achieved in Botswana. The WHA Resolution advocated for the control of schistosomiasis morbidity in highly endemic areas and urged countries to attain a target of regular treatment of “at least 75% and up to 100% of all school-aged children at risk of morbidity by 2010” [31]. The schistosomiasis program was terminated when it became clear that prevalence of the parasite remained below 10% and intestinal schistosomiasis was no longer a public health problem in the district [4]; thus, surveillance and monitoring were halted. Some studies have argued that schistosomiasis and other soil transmitted helminths are neglected diseases which are perceived to have a very low burden of disease index. Globally, more than a third of the world’s population are currently infected and children suffer from profound physical deficits such as anemia, malnutrition, stunted growth, and cognitive delays [32].

This study revealed a knowledge deficit, about 58% of children had never heard of bilharzia even though they lived in communities where the disease was previously endemic. In fact, with the termination of the program, the health training institutions and the Ministry of Education excluded NTDs in their curricula. The knowledge deficit was observed among the school age children at different grade levels. The lower grades learners’ knowledge of schistosomiasis was lowest and increased with grades. This is because health education was more emphasized at higher grades than lower ones. The finding is not surprising as similar results were reported in a Swaziland study, where knowledge discrepancies regarding schistosomiasis among primary school children were revealed [33]. The renewed attention on NTDs by WHO in 2017 is timely for countries like Botswana to curb morbidity that may result from the endemic and chronic nature of schistosomiasis [34]. Another interesting finding in this study is that learners who had a family member who previously suffered schistosomiasis had higher knowledge than those who did not. This is in line with other studies which showed that learners from areas (e.g., schools) with high prevalence of schistosomiasis showed higher knowledge of the disease and took preventive measures against reinfection [35]. Another study conducted among children in Yemen also found that children from families with history of schistosomiasis infections had higher knowledge than those who did not [36]. This may be the case when family members are infected with schistosomiasis, they learn about the disease.

The results also showed knowledge and history of disease within family as determinants of schistosomiasis preventive behaviors. It is surprising that learners with higher knowledge of schistosomiasis are likely to engage in risky behaviors compared to those with lower knowledge of the disease. Though surprising, previous studies in health behavior have shown that knowledge on its own does not lead to desired health behavior [3739]. The finding indicates that knowledge alone may not predict behavior as there are multiple factors that are needed to promote pro-health behaviors. Environment, peers, cues to action and situational factors can also play a role. It is therefore necessary that billboards are placed next to stagnant water pools as cues and public swimming facilities. Okavango sub-district does not have recreational facilities such as public swimming pools where adolescents may relax. Providing knowledge without alternatives may not be effective on its own.

In this study, an overall prevalence of S.hematobium and S.mansoni was 8.7% and 0.64% respectively. In addition, the results demonstrated that the intensity of schistosomiasis was generally light at 97.6% and heavy intensity at 2.4%; this calls for continued monitoring of morbidity and that treatment is required.

It is possible that the prevalence of schistosomiasis was higher in the upper and mid sections of the Okavango Delta because, during data collection the Okavango River had more water in Kauxwi, Shakawe and Etsha 6 than in Maun, the distal part. Children in the upper and mid Okavango were observed swimming in the river, as such, an association between swimming in infected water and the risk of infection was established. A lot of children were observed swimming in the two communities which seemed to predispose them to infection [40]. They attributed the hyperendemicity of the disease in the two communities to be intensified by water contact activities such as bathing, fishing, and swimming in cercariae infested streams, rivers, and ponds.

In Maun, the results might have been influenced by the drought spell the country experienced, more specifically during data collection, the Thamalakane River water level was very low, and this is the river which runs through this urban village. This could explain the low infection transmission and low prevalence rates. In addition, the study findings demonstrate that while S. Mansoni might be on the decline, S. haematobium is on the rise, which makes it a public health concern. Mass drug administration (MDA) with praziquantel was conducted in all schools where children tested positive, but this should be repeated to make sure treatment is effective. The current WHO recommendation is that in areas with a prevalence over 10% but less than 50% MDA should be conducted once every two years to be able to break and sustain the control of schistosomiasis transmission. We recommend that all school age children 6–13 years in the Okavango Delta should receive praziquantel once every two years to sustain the transmission of schistosomiasis. Mass drug administration should be carried out together with other schistosomiasis control strategies such as snail control and health education to reach elimination goals. In 2017, the World Health Assembly adopted resolution [34], calling member states to develop or adapt national vector control strategies and operational plans aimed at reducing by at least 40% the incidence of vector borne diseases, including schistosomiasis by 2025. WHO reinforces snail control as part of its strategic approach to eliminate schistosomiasis as a public health problem.

Ngamiland district like other districts in Botswana, still has limited access to safe drinking water (running water and protected wells or rivers) and sanitation (flush toilets and pit latrines). In this study, almost half (49%) fetched water from the river. The study further revealed that the main source of portable water which are community standpipes were dry most of the time, further putting these communities at risk for WASH related diseases. This use of pit latrines and open defecation which is still practiced has the potential of contaminating water sources. To control and prevent transmission of schistosomiasis, public health education is critical and effective measures should be put in place to improve water supply, sanitation, and hygiene infrastructure. In addition, public health interventions that focus on behavioral change and WASH measures may enhance effects of MDA against schistosomiasis and STH [41].

4.1 Limitations of study

This study had some limitations, the single screening of urine and stool specimens. Most studies recommend collection of samples on two consecutive days from the children to optimize recovery of eggs and better estimate infection level and true prevalence in those communities [40, 42]. Secondly, in some areas the processing site of samples was too distant, and we are not sure that this could have in anyway affected urine specimen. Thirdly, the study was conducted when the Delta was very dry hence low prevalence rates. Lastly, the research team could not repeat the mass drug administration to children who had tested positive because of the Covid-19 movement restriction.

5. Conclusions

Our study indicated an overall prevalence of SS. hematobium and S.mansoni at 8.7% and 0.64% respectively. Intensity of SS. hematobium was generally light (97.6%) and heavy intensity (2.4%). The findings also revealed a knowledge deficit, about 58% of children had never heard of bilharzia even though they lived in communities where the disease was previously endemic. Learners who had a family member who previously suffered from schistosomiasis had higher knowledge than those who did not. Interestingly, these learners were likely to engage in risky behaviors compared to those with lower knowledge of the disease. An integrated approach including health education, mass drug administration, surveillance, water, sanitation, and hygiene infrastructure should be prioritized for the prevention and control of schistosomiasis in the Botswana national NTD strategy. There is also an urgent need to undertake a large-scale study to determine schistosomiasis prevalence nationwide.

Supporting information

S1 File

(SAV)

Acknowledgments

We thank the Ministry of Basic Education for availing the primary schools in Ngamiland; the Ministry of Health and Wellness for availing their laboratory facilities as well as provide praziquantel treatment; the Community Leadership in the study sites, the community, and parents for allowing their children to participate in the study. Our heartfelt gratitude to the study participants, our research assistants, and our postgraduate students. We would also like to extend our gratitude to Mr. Kesaobaka Molebatsi for his support with statistical data analysis.

Data Availability

All relevant data are within the paper and its Supporting Information files.

Funding Statement

Funding information has been removed from the Acknowledgments section. We would like to maintain the Funding Statement as it reads "This study was funded by the National Institute of Health Research (Grant Number: 16/136/33 NIHR). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.” Any additional information provided contrary to the original statement should be deleted.

References

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Decision Letter 0

Clement Ameh Yaro

7 Dec 2022

PONE-D-22-26018The prevalence of schistosomiasis among primary school aged children (6-13 years) in the Okavango Delta in Botswana.PLOS ONE

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Reviewer #2: Yes

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Reviewer #1: Reviewer’s Comments________________________________________

Title: The prevalence of schistosomiasis among primary school aged children (6-13 years) in the Okavango Delta in Botswana

Manuscript Number: PONE-D-22-26018

Overall comments

The paper is of interest to schistosomiasis control program of Botswana as it provides information on re-emerging of the disease after the termination of the Botswana national schistosomiasis control program in 1993. Had the snail survey been conducted along the parasitological survey, the study would have more informed the schistosomiasis control program of the country.

The primary objective/purpose of this study was to determine the prevalence and intensity of schistosomiasis among the study participants. The title also reflects this to some extent. The paper should also be structured accordingly in all sections, i.e., presenting prevalence and intensity first, followed by participants’ knowledge of schistosomiasis, and so on.

Although the paper reports on important information it is not well structured and written. Hence, it needs substantial revision to bring it to the acceptable level.

Specific comments

Title:

It is suggested that the term “schistosomiasis” in the title be replaced with “intestinal and urogenital schistosomiasis”. Additionally, the authors need to replace the phrase “...primary school aged children (6-13 years)...” in the title and throughout in the document be replaced with “...school-age children...”

Abstract:

� Page 1,Background: provide purpose/objective of the study

� Page 1,Methods: more details required for methods used for stool and urine examination, as well as for statistical analysis of the data.

� Page 3, lines 46- 48: Recommendation should be preceded by summary of the main findings.

Page 7: It would be appropriate to create the heading “Study area and population” with which the method section begins.

Page 7, lines 118 - 121: It is not necessary to outline specific objectives in an article as it is not a proposal. It is usual to provide purpose /objective statement only.

Page 9, lines 164 & 165: For individuals who have not reached the legal age of consent, you only need consent from parents or legal guardians. Individuals who have not reached the legal age of consent are not expected to complete the assent form but only give oral assent.

Page 10, lines 183 & 184: The statement that reads “Total Urine (n=1603) and stool (n=1404) samples collected from school aged children from all the selected schools to determine the prevalence and intensity of schistosomiasis.” lacks clarity and needs to be re-phrased to read “A total of 1603 urine and 1404 stool samples were collected from school age children in the selected schools to determine the prevalence and intensity of schistosomiasis.” This is just one example, otherwise there are many other such vague statements for which the paper needs considerable editorial revision.

Page 11, lines 185 & 186: For examination of S. haematobium infection, WHO recommends urine filtration method. It is not clear why the authors used additional centrifugation. The authors need to clarify this. The authors also need to re-write the urine and stool examination methods and cite appropriate references.

Page 14, Table 2: vertical lines in the table should be removed.

Page 19, Table 5:

� The figures in column “S. haematobium” add up to 83, and not to 156.

� It is not clear why “Ascaris/Taenia” is in the table.

� Show STH identified in the footnote.

Page 20, Table 6:

� It is not clear why “Ascaris/Taenia” is in the table.

� Show STH identified in the footnote.

Page 21, Table 7: The number of S. haematobium infected cases were 86(83 in Table 5 & 3 in Table 6). On the other hand, the authors have written the number of S. haematobium infected cases as 41. The discrepancy should be reconciled.

Page 22, Table 8: The number infected with S. mansoni was 3 while the intensity of infection was presented for 1. Here correction is needed.

Page 24, line 412: “S. Haematobium” on this line and in other section should be corrected as “S. haematobium”

Page 27, lines 464 & 465: The statement “Our data provide baseline information regarding infection status of Schistosoma and STH among school aged children living along the Okavango Delta.” is a significant statement and should be replaced with a summary of the findings.

Reviewer #2: The manuscript provides interesting data at a time when the activities on Neglected Tropical Diseases is aiming for a global elimination. Such situation and existing conditions require to be highlighted. The data provides vividly that there are missing gaps in the whole approach in the road to elimination as depicted by the situiation in the Okavango region. This may not be an isolated existing condition but could be prevailing in other regions in the sub Saharan region. I have identified some sections that require proper presentation in conformity to the generally required for publications.

1. I suggest that the figures given in all the tables should have an integer for example where it is given as .40 % it should be 0.40%. This suggestion is for all the presnetation in the manuscript.

2. Line 66 the reference need to be written in a proper way instead of the listed names. Usually it is the only first author as (Hajissa et al., 2018)

3. Line 85 should read as : Other studies conducted in Ethiopia...

4. line 87: Conducted in Zimbabwe and Kenya....

5. line 183: Total urine (n=1603 and stool ...

6. line 187: ova indicated a positive diagnosis of S. heamatobium ...

7. The methods should not be listed as points but contained in a discriptive way.

8. Lines 349 and 359 the Genus and species names: S. heamatobium and S. mansoni..

9. Line 390 grammar: who had a family member who previously suffered ....

10. Remove information between 409 - 422 as it is repetition of lines 394 -396.

11. Line 412: S. haematobium

12. Lines 433 and 436: reconstruction of the sentence. sustain control of schistosomiasis transmission...

13. Line 461. COVID-19 restriction

References section require proper presentation.

Ref 506, 509: The author is the same Appleton CC or Appleton C?

ref 527: Presentation of the author is not clear if, B Doumbo,OK

Care should be taken in the use of reference manager where the format has changed fromn Ref 593 - 626.

**********

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Reviewer #1: Yes: Berhanu Erko

Reviewer #2: Yes: Professor Takafira Mduluza

**********

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PLoS One. 2023 May 30;18(5):e0285977. doi: 10.1371/journal.pone.0285977.r003

Author response to Decision Letter 0


16 Mar 2023

We are pleased to resubmit a manuscript PONE-D-22-26018 The prevalence of urogenital and intestinal schistosomiasis among school age children (6-13 years) in the Okavango Delta in Botswana by Nthabiseng A. Phaladze (Principal investigator), and co-authors Lebotse Molefi, Olekae T. Thakadu, Onalenna Tsima, Barbara N. Ngwenya, Tuduetso L. Molefi and Wananani B. Tshiamo for consideration to be published in PLOS One.

We wish to take this opportunity to heartily thank the reviewers for their valuable time and useful contribution to this work. We appreciate the thorough inputs they have given which will no doubt help improve our manuscript. The following paragraphs contain point-by-point responses to the reviewers’ and editorial board comments:

A. Response on Journal Requirements:

1. Compliance of the Manuscript to PLOS ONE’s style requirements: We are happy to submit the revised manuscript in line with all the requirements of the journal.

2. Competing Interests Statement: I declare that the authors have no conflict of interest. Mr . Lebotse Molefi is a paid employee of Quality Anchor Consultants PTY. LTD, but was not affiliated with the study at the time of this study was conducted. There are no patents, products in development or marketed products associated with this research to declare. This does not alter our adherence to PLOS ONE policies on sharing data and materials.”

3. Funder: Funding information has been removed from the Acknowledgments section. We would like to maintain the Funding Statement as it reads "This study was funded by the National Institute of Health Research (Grant Number: 16/136/33 NIHR). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.” Any additional information provided contrary to the original statement should be deleted.

4. Copyright of Figures 1 & 2: have been deleted as advised since it was difficult to get permission from the copyright holder.

B. Comments to the Author:

1. Is the manuscript technically sound, and do the data support the conclusions?

We note that Reviewers agree on this aspect. We have also noted comments made by Reviewer #1.

2. Has the statistical analysis been performed appropriately and rigorously?

We note that Reviewers agree on the statistical analysis.

3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

We also note that Reviewers agree on this aspect.

4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in

submitted articles must be clear, correct, and unambiguous. Any

typographical or grammatical errors should be corrected at revision, so please

note any specific errors here.

We do note the Reviewers divergent views on this question; but do take comments made by Reviewer #1 seriously and have attended to typographical and grammatical errors.

5. Review Comments to the Author:

a. Reviewer #1

1. Title: The suggestion to change the title to “The prevalence of urogenital and intestinal schistosomiasis among school age children (6-13 years) in the Okavango Delta in Botswana” has been adopted.

The phrase “primary school aged children (6-13 years)” in the title and throughout in the document has been replaced with “school-age children in the title and throughout the text.

2. Abstract: Page 1, Background: line 1, the study purpose has been provided “This study sought to investigate prevalence and intensity of schistosomiasis among school age children 6-13 years in selected communities in the Okavango Delta”.

- Methods: More details of methods used for stool and urine examination, as well as for statistical analysis of data have been provided.

- Recommendation preceded by main findings as per advice.

3. Page 7, Specific objectives have been removed, and the study purpose maintained.

4. Page 7, Heading created “Study area and population”.

5. Ethical considerations: statement on assent by children corrected on page 9, lines 156-157.

6. The statement that reads “Total Urine (n=1603) and stool (n=1404) samples collected from school aged children from all the selected schools to determine the prevalence and intensity of schistosomiasis.” was re-phrased as suggested to read “A total of 1603 urine and 1404 stool samples were collected from school age children in the selected schools to determine the prevalence and intensity of schistosomiasis.” More details on methods used to analyze urine and stool samples are provided on Page 10.

7. Comment: For examination of S. haematobium infection, WHO recommends urine filtration method. It is not clear why the authors used additional centrifugation. The authors need to clarify this.

- Clarification: The majority of intestinal, urinary and blood parasites

can be detected microscopically in unstained or stained preparations,

either directly or following concentration by centrifugation. (Medical

Laboratory Manual for Tropical Countries Volume 1 page 178 (second

edition by Monica Cheesbrough (ELBS-English Language Book

Society/Tropical Health Technology/ 1987).

To increase the sensitivity of detecting parasite ova, urine filtration

and centrifugation methods were used to analyze urine samples

concurrently. Positive diagnosis of S. haematobium was based on the

detection of one terminal spined schistosoma ovum or more.

8. The urine and stool examination methods have been re-written and appropriate references cited.

9. Comments on Tables 2;5;6;7 &8 have been addressed and statistical corrections done. The issue of other soil transmitted helminths -Ascaris, Taenia have been removed from the tables. PLOS ONE does not allow footnotes. We added a sentence in the text to acknowledge that even though other STHs were not the focus, during urine and stool examination, they were identified.

10. The statement “Our data provide baseline information regarding infection status of Schistosoma and STH among school aged children living along the Okavango Delta.” The statement has been replaced with a summary of the findings.

b. Reviewer #2:

1. Integer inserted in all Figures given in all the tables and throughout the

manuscript.

2. Line 66: Referencing of Hajissa et al., 2018 corrected.

3. Lines 85; 87; 183 & 187 corrected as per advice.

4. Methods of examination for urine and stool samples written in a

narrative form; bullets removed.

5. Lines 349, 359 and 412: Genus and species names corrected as per

advice.

6. Repetition of information removed, and grammar corrected.

7. Lines 433 and 436: Reconstruction of the sentence corrected as

suggested.

8. Line 461: Covid-19 restriction accepted.

9. References Section corrected.

We hope these responses meet your expectations and awaiting your decision.

Sincerely,

Nthabiseng A. Phaladze PhD.

Professor of Nursing

University of Botswana School of Nursing

Email: phaladze@ub.ac.bw

Alternate email: nphaladze@yahoo.co.uk

Attachment

Submitted filename: Response to Reviewers_PONE-D-22-26018 JANUARY 20 2023.docx

Decision Letter 1

Clement Ameh Yaro

7 May 2023

The prevalence of urogenital and intestinal schistosomiasis among school age children (6-13 years) in the Okavango Delta in Botswana.

PONE-D-22-26018R1

Dear Dr. Phaladze,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.

If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Clement Ameh Yaro, Ph.D

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: (No Response)

**********

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If you choose “no”, your identity will remain anonymous but your review may still be made public.

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Reviewer #1: Yes: Berhanu Erko

**********

<quillbot-extension-portal></quillbot-extension-portal>

Acceptance letter

Clement Ameh Yaro

16 May 2023

PONE-D-22-26018R1

The prevalence of urogenital and intestinal schistosomiasis among school age children (6-13 years) in the Okavango Delta in Botswana.

Dear Dr. Phaladze:

I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department.

If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org.

If we can help with anything else, please email us at plosone@plos.org.

Thank you for submitting your work to PLOS ONE and supporting open access.

Kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Dr. Clement Ameh Yaro

Academic Editor

PLOS ONE

Associated Data

    This section collects any data citations, data availability statements, or supplementary materials included in this article.

    Supplementary Materials

    S1 File

    (SAV)

    Attachment

    Submitted filename: Response to Reviewers _PONE-D-20-35623 July 2021.docx

    Attachment

    Submitted filename: Response to Reviewers_PONE-D-22-26018 JANUARY 20 2023.docx

    Data Availability Statement

    All relevant data are within the paper and its Supporting Information files.


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