Detection of Adenoviruses and Astroviruses in Patients and Marine Animals in the Republic of Guinea

Detection of Adenoviruses and Astroviruses in Patients and Marine Animals in the Republic of Guinea | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL This is a preprint and has not been peer reviewed. Data may be preliminary. 14 May 2025 V1 Latest version Share on Detection of Adenoviruses and Astroviruses in Patients and Marine Animals in the Republic of Guinea Authors : André Saa TOGBODOUNO 0009-0005-8211-3811 [email protected] , Roland TENKIANO , Emmanuel Saa MILLIMONO , René Tamba Tolno , Jacqueline Sia Mara , Ramatoulaye BALDE , Lansana II SOUMAH , Moussa KOLIÉ , Sanaba Boumbaly , Boubacar Sidy Sily BAH , and Mohamed Sahar TRAORE Authors Info & Affiliations https://doi.org/10.22541/au.174720075.52390810/v1 925 views 128 downloads Contents Abstract Abstract Supplementary Material Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract Introduction: Gastroenteritis is a major public health concern, particularly in low-income countries. Adenovirus types 40 and 41 are associated with diarrhoea in humans. Approximately 1.7 billion cases of diarrhoea occur each year, with over 525,000 deaths among young children globally. This study aims to determine the prevalence of adenoviruses and astroviruses in patients and marine animals in the Republic of Guinea. Methods: Biological samples were collected from three localities (Kindia, Gueckédou, and Conakry) and analysed using qPCR and RT-PCR. Results were examined based on the type of diarrhoea, age, gender, season, and socio-professional category. Statistical analysis was performed using the z-test to evaluate the significance of observed differences. Results: Positivity rates varied by location: Kindia showed the highest prevalence, particularly in liquid diarrhoea (7.41%). Adenoviruses were more frequent (33.23% of 250 samples), while astroviruses were detected in only 0.71% of cases. Adenovirus prevalence was higher during the rainy season and among children (8.19%), followed by young adults (5.26%). Differences based on gender and socio-professional categories were not statistically significant (p=0.52). One case of astrovirus (out of 40 samples) was detected in marine animals. Conclusion: The study highlights a significant circulation of adenoviruses, especially among children and during the rainy season. The results suggest that environmental and sanitary factors may influence transmission. Authors: André Saa TOGBODOUNO 1* , Roland TENKIANO 1 , Emmanuel Saa MILLIMONO 1 , René Tamba Tolno 1 , Jacqueline Sia Mara 4 , Ramatoulaye BALDE 1 , Lansana II SOUMAH 1 , Ibrahima Sory SOW1,Moussa KOLIÉ 5 ,Sanaba Boumbaly 3 , Boubacar Sidy Sily BAH 2 , Mohamed Sahar TRAORE 1 . Affiliations: 1 Institute of Applied Biology Research of Guinea 2 University of Kindia 3 Virology Research Centre 4 Regional Hospital of Kindia 5 Prefectural Hospital of Gueckédou *Corresponding Author: [email protected] | Phone: +224 620 03 33 79 /Orcid: https://orcid.org/0009-0005-8211-3811 Director: [email protected] | Phone: +224 628 52 92 04 Author Contributions André Saa TOGBODOUNO: Sample collection, transportation from Gueckédou to the laboratory, and manuscript writing. Lansana II SOUMAH: Development of the research protocol. Aboubacar Sidy Sily BAH: Revision and validation of the research protocol. Emmanuel Saa MILLIMOUNO, Tamba René Tolno, and Moussa KOLIÉ: First round of manuscript revision. Mohamed Sahar TRAORE: Critical review and final editing of the manuscript. Ibrahima Sory SOW: Statistical data analysis. Roland TENKIANO: Sample collection in Kindia and Conakry. Jacqueline Sia Mara: Sample preprocessing. Ramatoulaye BALDE and Sanaba Boumbaly: Molecular analysis of samples. Abstract Introduction: Gastroenteritis is a major public health concern, particularly in low-income countries. Adenovirus types 40 and 41 are associated with diarrhoea in humans. Approximately 1.7 billion cases of diarrhoea occur each year, with over 525,000 deaths among young children globally. This study aims to determine the prevalence of adenoviruses and astroviruses in patients and marine animals in the Republic of Guinea. Methods: Biological samples were collected from three localities (Kindia, Gueckédou, and Conakry) and analysed using qPCR and RT-PCR. Results were examined based on the type of diarrhoea, age, gender, season, and socio-professional category. Statistical analysis was performed using the z-test to evaluate the significance of observed differences. Results: Positivity rates varied by location: Kindia showed the highest prevalence, particularly in liquid diarrhoea (7.41%). Adenoviruses were more frequent (33.23% of 250 samples), while astroviruses were detected in only 0.71% of cases. Adenovirus prevalence was higher during the rainy season and among children (8.19%), followed by young adults (5.26%). Differences based on gender and socio-professional categories were not statistically significant (p=0.52). One case of astrovirus (out of 40 samples) was detected in marine animals. Conclusion: The study highlights a significant circulation of adenoviruses, especially among children and during the rainy season. The results suggest that environmental and sanitary factors may influence transmission. Keywords: Adenovirus, Astrovirus, Diarrhoea, Prevalence, Guinea Acute gastroenteritis represents a major public health concern, particularly in developing countries. Human adenoviruses (HAdVs) and astroviruses (HAstVs) are responsible for approximately 70% of cases, primarily affecting children under the age of five (Hartman RM, 2023). Adenoviruses are double-stranded DNA viruses, with serotypes 40 and 41 specifically associated with diarrhoeal diseases in humans (Ghebremedhin, 2014). Globally, an estimated 1.7 billion cases of childhood diarrhoea occur annually, resulting in approximately 525,000 deaths among children under five years old (WHO, 2023), particularly in sub-Saharan Africa and South Asia (Dadonaite B, 2023). Enteric viruses such as HAdVs and HAstVs are thought to cause over 200,000 human deaths annually (Stuempfig ND, 2023). Astroviruses, which are single-stranded RNA viruses, are also a major cause of diarrhoea, particularly among young children and immunocompromised individuals (Dennehy PH, 2001; Troeger C., 2018). These pathogens are frequently implicated in both outbreaks and sporadic cases. In Guinea, specific detection of HAdVs and HAstVs is crucial for improving patient management and guiding epidemiological investigations. Nevertheless, despite advanced diagnostic techniques, a significant proportion of gastroenteritis cases remain unexplained (X. Lu et al., 2006; Yinda CK, 2019). This study aims to evaluate the prevalence of human adenoviruses and astroviruses in patients presenting with diarrhoea and vomiting, as well as in marine animals. Polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR) techniques were employed to detect astroviruses. The study seeks to enhance local epidemiological understanding and inform strategies to combat diarrhoeal diseases in the hospitals of Gueckédou, Kindia, and the marine environment of Conakry, and more broadly throughout Guinea. 2. Materials 2.1. Study Area Conakry, the capital city of the Republic of Guinea in West Africa, is located on the Kaloum Peninsula and bordered by the Atlantic Ocean and surrounding islands. The city features diverse topography, including coastal plains and hills, and is predominantly covered by mangrove vegetation. Its tropical climate is characterised by average temperatures ranging from 24.6°C to 37.5°C, with heavy rainfall, particularly during July and August. Covering an area of 450 km², Conakry had a population of over 1.66 million in 2014, composed mainly of young people from the Soussou ethnic group, known for its rich cultural traditions and folklore. The city is administratively divided into 13 autonomous municipalities governed by a central governorate (MATD, 2024, matd.gov.gn). Kindia, on the other hand, holds a strategic geographic position and benefits from significant natural resources. Spanning 500 km², it is surrounded by plateaus averaging 400 m in elevation, with peaks reaching up to 1,117 m. The region is well-irrigated, with an extensive hydrographic network. Its humid tropical climate alternates between dry and rainy seasons, with temperatures ranging from 15°C to 38°C. In 2018, Kindia had an estimated population of around 296,000, composed primarily of youth and women from diverse ethnic backgrounds. The local economy is primarily based on agriculture and livestock farming (RGPH-3, 2014, https://www.stat-guinee.org). Lastly, the prefecture of Guéckédou, located in the forested region of Guinea, spans a vast area of 4,400 km². As of 2016, its population was estimated at approximately 610,000, predominantly from the Kissi ethnic group. The region is intersected by numerous watercourses and features a savannah climate, with an annual average temperature of 26.4°C and significant summer rainfall. Rich in biodiversity and natural resources, Guéckédou serves as an important commercial hub, notably for rice, coffee, and palm-derived products (Köppen classification / Guinea / Guéckédou prefecture). 2.3. Sample Collection A total of 290 samples were collected for this study: 250 from patients aged between 0 and 65 years suffering from diarrhoea—100 from Kindia Regional Hospital and 150 from Guéckédou Prefectural Hospital—and 40 from marine animals in Conakry (comprising 19 fish and 21 crabs). The collection process complied with the ethical guidelines for research in Guinea and was conducted with informed consent. Each sample was accompanied by a detailed data sheet documenting the participant’s identity, age, sex, symptoms, and date of collection. Samples were sequentially numbered, with between 6 and 14 samples collected daily. 2.4. Transport Samples were stored in 1.5 mL Eppendorf tubes and kept in cool boxes containing dry ice. They were transported within 72 hours to the Guinean-Russian Laboratory at the Institute of Applied Biology Research of Guinea (IRBAG), where the analyses were conducted. 3. Methodology 3.1. Sampling This six-month study (from 5 July to 5 December 2022) was based on a random sampling approach. It was designed as a prospective and cross-sectional investigation targeting patients aged 0 to 65 years from Guéckédou Prefectural Hospital, Kindia Regional Hospital, and marine animals. 3.2. Inclusion Criteria All patients aged 0–65 years presenting with symptoms such as diarrhoea or vomiting who gave informed consent were included in the study. 3.3. Study Variables Variables included patients and marine animals, age, sex, socio-professional status, origin of patients, and sampling location. 3.4. Exclusion Criteria Patients aged 0–65 years presenting with symptoms but who did not provide informed consent were excluded from the study. 4. DNA Extraction of Human Adenoviruses and Astroviruses The extraction process followed the protocol of the Ribo-Sorb Rotor-Gene 6000 AmpliSens® kit. 4.1. Preparation Each 100 µL sample, including positive and negative controls, was mixed with 450 µL of lysis solution and incubated at 65°C for 15 minutes before centrifugation. An internal control (10 µL) was added, followed by further incubation at 65°C. Subsequently, 30 µL of absorbent solution was introduced and the mixture centrifuged at 13,000 rpm. 4.2. Washing Steps The washing procedure involved three steps: wash solution 1, wash solution 2 (70% ethanol), and wash solution 3 (acetone), each followed by centrifugation. After removing the supernatant, 50 µL of elution buffer was added. The extracted DNA was either immediately used for amplification or stored at -20°C. 4.3. Final Eluate Real-time PCR was performed using the AmpliSens® kit, which includes a hot-start polymerase to reduce non-specific priming. Each reaction comprised 15 µL of master mix combined with 10 µL of extracted DNA. Fluorescent dyes enabled real-time monitoring of amplification, with the method achieving a sensitivity threshold of 1×10⁴ copies for each targeted pathogen. 5. Data Analysis Data analysis was conducted using statistical methods including the calculation of the p-value, a key indicator in hypothesis-driven scientific research (Oliver Y. Chen et al., 2023). A 95% confidence interval (CI) and Microsoft Excel software were employed to process and interpret the collected data. 6. Results Figure 1: Characterisation and Origin of the Samples Figure 1 illustrates the positivity rates of samples collected from three regions: Kindia, Gueckédou, and Conakry. In Kindia, liquid diarrhoea exhibited a slightly higher positivity rate (7.41%) compared to bloody diarrhoea (6.85%). In Gueckédou, both rates were lower (3.41% and 3.23% respectively), indicating reduced pathogen circulation in the area. In Conakry, samples collected from marine animals showed a low positivity rate (2.50%). Overall, Kindia recorded the highest level of contamination, and liquid diarrhoea appeared more frequently associated with positive cases. Table 1: Distribution of samples according to the pathogens studied. Adenovirus 250 12 32.52 [2,15; 7,45] 0,514 Astrovirus 40 1 0,71 [0; 7,34] Total 290 13 33.23 [2.10; 6.86] - From Table 1, it appears that adenoviruses are the most frequently detected, with 250 samples tested and 12 positive results (33.23%). Astrovirus was detected in 1 out of 40 samples (0.71%). The confidence intervals reflect potential variability in actual infection rates. A statistical comparison using the z-test yielded a p-value of 0.514 (> 0.05), indicating that the difference in detection rates between the two viruses is not statistically significant. Table 2: Samples by Age Group 1 0 – 15 122 10 8.19 [3.33 -13.06] 2 16 – 30 78 1 1.28 [0 - 3.78] 3 31– 45 19 1 5.26 [0 -15.30] 4 46− 65 31 0 0 [0 - 0] Total 250 12 4.8 [0.26 - 3.74] This reveals that among the 250 samples tested, the overall positivity rate was 4.8%. The highest rate was observed in the 0–15 years age group (8.19%), followed by the 31–45 years group (5.26%). The 16–30 years group showed a rate of 1.28%, while no positive cases were detected among individuals aged 46–65. These findings suggest an age-related variation in positivity, with higher prevalence among younger individuals. Table 3: Distribution of Positive Cases by Sex 1 Female 167 7 4.19 [0.01%, 7.23%] 0.52 2 Male 83 5 6.02 [0.91%, 11.14%] Total 250 12 10.21 [2.15%, 7.45%] From Table 3, positivity rates between males (6.02%) and females (4.19%) appear slightly different. However, the p-value of 0.52 far exceeds the conventional significance threshold (0.05), suggesting no statistically significant difference in infection rates between sexes. The overlapping 95% confidence intervals reinforce this interpretation. The small difference in percentages likely results from sample size variation rather than true difference in prevalence. Table IV: Distribution of Results by Season 1 Dry season 81 2 1.62 [-0.91- 5.85 ] 2 Rainy season 169 10 16.9 [2.36 - 9.47 ] Total 250 12 4.8 [2.15 - 7.45 ] Table IV shows that the prevalence of positive cases was higher during the rainy season (16.9%) compared to the dry season (1.62%). The wide and null-inclusive confidence interval for the dry season reflects uncertainty, likely due to the low number of positive cases. These results suggest seasonal variation, with higher transmission during periods of increased humidity. Table V: Socio-Professional Distribution of Human Adenovirus Positive Cases Drivers/Labourers 1 0 0.00% [0.0 _ 79.35] 1 Civil Servants 29 1 3.45% [0.61 _17.18] 0.463 Traders 42 0 0.00% [0.0_8.38] 0.020 Pupils/Students 11 1 9.09% [1.62_37.74] 1 Housewives 18 1 5.56% [0.99_25.76] 1 Farmers 8 0 0.00% [0.0 _32.44] 1 Others (Children) 141 9 6.38% [3.39_11.69] 0.025 Total 250 12 4.80% [2.77_8.20] 0.68 Analysis of Table V shows that housewives (5.56%), students (9.09%), civil servants (3.45%), and children (6.38%) were among the groups with positive cases reported in the cities of Gueckédou and Kindia. The p-value of 0.68 exceeds the standard significance level, indicating no statistically significant association between occupational category and positive results. Differences in infection rates may be attributed to factors such as healthcare access, hygiene practices, or education. Table 6: Distribution of Seafood Product Samples 1 Fish 21 0 0 [0 ; 15,46 ] 2 Crabs 11 0 0 [0 ; 25,88 ] 3 Others (molluscs) 8 1 12,5 [2,24 ; 47,09 ] Total 40 1 2,5 [0,44 ; 12,88 ] According to Table VI, none of the fish or crab samples tested positive, while one positive case (12.5%) was detected among other marine animals (molluscs). Although the majority of tests were negative, this does not equate to a zero-risk scenario. The presence of a single positive case highlights a potential, albeit low, risk of virus transmission in marine animals. 7. Discussion The study on the detection of adenoviruses and astroviruses in patients and marine animals in the Republic of Guinea provides evidence for the circulation of these viruses, based on case distribution across various variables and associated risk factors. Positivity rates varied across the sites of Kindia, Gueckédou, and Conakry. In Kindia, watery diarrhoea (7.41%) showed a slightly higher positivity rate compared to bloody diarrhoea (6.85%). This may indicate that cases of watery diarrhoea are more frequently associated with the presence of adenoviruses or astroviruses, although other clinical or environmental factors may also contribute. In contrast, lower rates in Gueckédou suggest reduced circulation of these viral agents in that region. Marine animal testing in Guinea yielded a low positivity rate (2.50%), suggesting limited exposure or low transmission frequency of these viruses among marine species. In terms of frequency, adenoviruses were the predominant pathogen, with 33.23% positive samples out of 250 tested, compared to only 0.71% for astroviruses. While this difference suggests higher prevalence, the z-test yielded a p-value of 0.514, indicating no statistically significant difference between the detection rates (Table I). These findings are consistent with reports from other regions, such as South-East Asia and Latin America (Mousavi Nasab, 2020), where adenoviruses are also more common than astroviruses. Age-based analysis shows the highest positivity in children aged 0–15 years (8.19%), followed by young adults aged 26–45 (5.26%), with much lower rates in other age groups (Table II). This suggests increased susceptibility in younger children, consistent with findings from other studies in low-resource settings, where children under five are most affected by viral gastroenteritis (Maksimov, 2015). This may reflect immature or absent immunity in this demographic. Although the positivity rates for males (6.02%) and females (4.19%) differ slightly, the p-value of 0.523 confirms no significant difference. This aligns with previous studies that reported no gender-based difference in virus prevalence (e.g., Hoffmann, 2018). The confidence intervals include the null value, suggesting that the observed low prevalence may reflect limited viral circulation. Students and children showed higher positivity rates (around 9%), potentially due to close social contact. However, this was not supported by statistical analysis (p > 0.05), implying no significant association between socio-professional category and infection. Other factors such as healthcare access, hygiene, and awareness may be more relevant. Regarding marine animals, the absence of positive cases in fish and crabs may indicate low prevalence, yet the single positive case among molluscs signals that the risk of zoonotic or foodborne transmission, though low, cannot be entirely ruled out (Table VI). This aligns with other studies on animal reservoirs of enteric viruses, which report rare but possible interspecies transmission (Griffiths, 2013). These findings are broadly consistent with research from other regions (e.g., Hoffmann, 2018), although environmental and social factors vary by context. 8. Conclusion This study highlights the role of environmental conditions particularly humidity and temperature in the frequency of gastrointestinal infections, with higher rates during the rainy season. Although certain groups, such as children and students, showed increased positivity, no statistically significant differences were observed, pointing instead to hygiene and healthcare access as key factors. The detection of a single positive case in marine animals suggests a low but non-negligible risk of interspecies transmission. These findings call for further research, particularly on seasonal dynamics, socio-economic determinants, and the inclusion of animal health surveillance within a ”One Health” framework. Abbreviations HAdV: Human Adenoviruses HAstV: Human Astroviruses PCR: Polymerase Chain Reaction RT-PCR: Reverse Transcription Polymerase Chain Reaction MATD: Ministry of Territorial Administration and Decentralisation RGPH-3: Third General Population and Housing Census IRBAG: Institute of Applied Biological Research of Guinea N: number of samples n: number of positive cases %: prevalence Funding This study was made possible through financial support from the Ministry of Higher Education, Scientific Research and Innovation of the Republic of Guinea, and the Directorate General of the Institute of Applied Biological Research of Guinea (IRBAG). Conflict of Interest The authors confirm that there are no conflicts of interest of any kind related to this study. Ethical Statement This study was conducted in accordance with current research ethics in the Republic of Guinea. The hospitals, sites visited, patients, and relevant authorities all granted approval for the study. 9. 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Gut virome analysis of Cameroonians reveals high diversity of enteric viruses, including potential interspecies transmitted viruses. mSphere, 4(1), e00585-18. https://doi.org/10.1128/mSphere.00585-18 Supplementary Material File (figure.doc) Download 38.00 KB File (tables.doc) Download 50.50 KB Information & Authors Information Version history V1 Version 1 14 May 2025 Copyright This work is licensed under a Non Exclusive No Reuse License. Keywords adenovirus astrovirus epidemiology fecal contaminant virus classification Authors Affiliations André Saa TOGBODOUNO 0009-0005-8211-3811 [email protected] Institute of Applied Biology Research of Guinea View all articles by this author Roland TENKIANO Institute of Applied Biology Research of Guinea View all articles by this author Emmanuel Saa MILLIMONO Institute of Applied Biology Research of Guinea View all articles by this author René Tamba Tolno Institute of Applied Biology Research of Guinea View all articles by this author Jacqueline Sia Mara Regional Hospital of Kindia View all articles by this author Ramatoulaye BALDE Institute of Applied Biology Research of Guinea View all articles by this author Lansana II SOUMAH Institute of Applied Biology Research of Guinea View all articles by this author Moussa KOLIÉ Prefectural Hospital of Gueckédou View all articles by this author Sanaba Boumbaly Virology Research Centre View all articles by this author Boubacar Sidy Sily BAH University of Kindia View all articles by this author Mohamed Sahar TRAORE Institute of Applied Biology Research of Guinea View all articles by this author Metrics & Citations Metrics Article Usage 925 views 128 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation André Saa TOGBODOUNO, Roland TENKIANO, Emmanuel Saa MILLIMONO, et al. 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