Genomic Surveillance of SARS-CoV-2: Data Analysis and Assessment of Tunisian Strategy from January 2021 to February 2022.

Due to the emergence of the SARS-CoV-2 B.1.1.7 (Alpha) variant in the UK in 2020 and its risk of increased transmission, the Ministry of Health in Tunisia implemented a sequencing surveillance strategy for SARS-CoV-2. The aim of this study was to analyze SARS-CoV-2 genomic surveillance data in Tunisia (January 2021-February 2022) and to assess the implementation of the sequencing strategy for SARS-CoV-2 in accordance with national recommendations and the guidance for SARS-CoV-2 genomic surveillance for public health goals. A descriptive study of all sequenced RT-PCR samples sequenced (January 2021-February2022). An internal audit was also done to assess the compliance against standards covering national recommendations and the Guidance for SARS-CoV-2 genomic surveillance for public health goals. A total of 12 simple or composite requirements related to the following areas were included in the audit standards: sampling (one requirements); data collection/analysis (six requirements); partnership (one requirement); and ethical considerations (one requirement). A total of 4819 samples were sent to laboratories and 4278 samples were sequenced. A total of 3648 samples were classified. Positive variants of concern (VOC) samples were 80.92%, differentiated as follows: Alpha, 40.24%; Beta, 0.24%; Gamma, 0.03%; Delta, 45.26%; and Omicron, 14.19%. Three principal phases of VOCs per ISO-week were shown: Alpha 3/2021-25/2021; Delta 26/2021-2/2022; and Omicron 3/2022-6/2022. Levels of compliance were identified; from a total of 12 requirements, 7 were considered as "not met", 4 as "partially met", and 1 as "fully met" but including not totally achieved objectives. In conclusion, the internal audit of the national SARS-CoV-2 sequencing strategy revealed an overall "not met" level of compliance. The results offered a trigger to collaborate with all stakeholders to develop a surveillance strategy for early detection and response to outbreaks caused by VOCs.

Outbreak Investigation of Typhoid Fever in the District of Gabes, South of Tunisia.

Typhoid fever is a significant public health concern in many parts of the world, particularly in developing countries with poor sanitation and hygiene conditions. In July 2016, an outbreak of typhoid fever occurred in Ghannouche, located in the south of Tunisia. This paper reports the results of a field investigation undertaken to identify possible transmission pathways and risk factors in order to propose control and preventive measures. A retrospective cohort study including a passive and active case finding, as well as an environmental and bacteriological investigation was conducted from July to September 2016. A case was defined as a person residing or having stayed in Ghannouche and having presented from the beginning of June clinical signs suggestive of typhoid fever, with, for a confirmed case, laboratory isolation of S.Tyhi, and for a probable case, an epidemiological link with a confirmed case. Attack rates were determined, and risk ratios were estimated with respect to exposures. Unadjusted and adjusted odds ratios were estimated using binary logistic regression. Among the 628 subjects investigated, 102 cases of typhoid fever were identified (74 confirmed and 28 probable) with an overall attack rate of 16.24%. Over 56% of cases were male and those under 10 years old were most affected (38.2%% of cases) with a median age of 12 years (interquartile range 5 to 25 years). The main clinical signs were fever (95%) and diarrhea (57%). Young age (adjusted OR = 0.95 and 95% CI = 0.93-0.97), low level of education (adjusted OR = 4.76 and 95% CI = 1.34-16.81), and the habitat type Arab or rudimentary house (adjusted OR = 4.93 and 95% CI = 2.61-8.27) were the socio-demographic factors independently associated with typhoid fever. Typhoid fever was found to be associated with drinking softened water (adjusted OR = 2.64 and 95% CI = 1.16-4.82), eating raw fruit and vegetables from family gardens (adjusted OR = 6.13 and 95% CI = 3.66-11.06), and using uncontrolled waste disposal (adjusted OR = 3.52 and 95% CI = 2.03-6.94). A total of 110 drinking water samples were analyzed; out of the 38 samples of softened water, 12 were non-compliant and 5 were positive for Salmonella. The screening activity identified two asymptomatic carriers, one of whom was a softened water seller. We concluded that drinking softened water from informal or unauthorized sale units, consuming fruit and vegetables from family gardens, uncontrolled dumping of household waste, and poor socio-economic conditions increase the risk of typhoid fever in this region. Many recommendations were implemented to stop this outbreak and to prevent further episodes.

Baseline Seroprevalence of SARS-CoV-2 Specific Antibodies in Hot Spot Areas of Great Tunis, up to 3 Months Post Disease Onset in Tunisia.

The extent of the SARS-CoV-2 circulation and the COVID-19 epidemic in Tunisia three months after virus circulation was unknown. The aim of this study was to determine the extent of SARS-CoV-2 infection among household contacts of confirmed COVID-19 cases living in Hot spot areas of Great Tunis, Tunisia by estimating the seroprevalence of antibodies anti SARS-CoV-2 and to identify factors associated to seroprevalence at the first stage of the pandemic in order to guide decision making and to constitute a baseline for further longitudinal analysis of protective immunity to SARS-CoV-2. The National Observatory of New and Emerging Diseases (ONMNE), Ministry of Health Tunisia (MoH), with the support of the Office of the World Health Organization Representative in Tunisia and the WHO Regional Office for the Eastern Mediterranean (EMRO)), conducted a household cross-sectional survey on April 2020 in Great Tunis (Tunis, Ariana, Manouba and Ben Arous). The study was based on the WHO seroepidemiological investigation protocol for SARS-CoV-2 infection. SARS-CoV-2 specific antibodies (IgG and IgM) were qualitatively detected using a lateral immunoassay that detect SARS-CoV-2 nucleocapsid protein and administered by the interviewers. The included subjects were confirmed COVID-19 cases and their households contacts resided in hot spot areas (cumulative incidence rate ≥ 10 cases/100,000 inhabitants) of Great Tunis. Results: In total, 1165 subjects were enrolled: 116 confirmed COVID-19 cases (43 active cases and 73 convalescents cases) and 1049 household contacts resided in 291 households. The median age of participants was 39.0 with 31 years' interquartile range (Min = 8 months; Max = 96 years). The sex ratio (M/F) was 0.98. Twenty-nine per cent of participants resided in Tunis. The global crude seroprevalence among household contacts was 2.5% (26/1049); 95% CI 1.6-3.6%, 4.8%; 95% CI 2.3-8.7% in Ariana governorate and 0.3%; 95% CI 0.01%-1.8% in Manouba governorate. In multivariate analysis, the associated factors independently related to seroprevalence were age ≥25 years (aOR = 5.1; 95% CI 1.2-22.0), history of travel outside Tunisia since January 2020 (aOR = 4.6; 95% CI 1.7-12.9), symptomatic illness in the previous four months (aOR = 3.5; 95% CI 1.4-9.0) and governorate of residence (p = 0.02). The low seroprevalence estimated among household contacts in Great Tunis reflect the effect of public health measures early taken (national lockdown, borders closed, remote work), the respect of non-pharmaceutical interventions and the efficacy of COVID-19 contact-tracing and case management at the first stage of the pandemic in Tunisia.

Phylogenetic and amino acid signature analysis of the SARS-CoV-2s lineages circulating in Tunisia.

Since the beginning of the Coronavirus disease-2019 pandemic, there has been a growing interest in exploring SARS-CoV-2 genetic variation to understand the origin and spread of the pandemic, improve diagnostic methods and develop the appropriate vaccines. The objective of this study was to identify the SARS-CoV-2s lineages circulating in Tunisia and to explore their amino acid signature in order to follow their genome dynamics. Whole genome sequencing and genetic analyses of fifty-eight SARS-CoV-2 samples collected during one-year between March 2020 and March 2021 from the National Influenza Center were performed using three sampling strategies.. Multiple lineage introductions were noted during the initial phase of the pandemic, including B.4, B.1.1, B.1.428.2, B.1.540 and B.1.1.189. Subsequently, lineages B1.160 (24.2%) and B1.177 (22.4%) were dominant throughout the year. The Alpha variant (B.1.1.7 lineage) was identified in February 2021 and firstly observed in the center of our country. In addition, A clear diversity of lineages was observed in the North of the country. A total of 335 mutations including 10 deletions were found. The SARS-CoV-2 proteins ORF1ab, Spike, ORF3a, and Nucleocapsid were observed as mutation hotspots with a mutation frequency exceeding 20%. The 2 most frequent mutations, D614G in S protein and P314L in Nsp12 appeared simultaneously and are often associated with increased viral infectivity. Interestingly, deletions in coding regions causing consequent deletions of amino acids and frame shifts were identified in NSP3, NSP6, S, E, ORF7a, ORF8 and N proteins. These findings contribute to define the COVID-19 outbreak in Tunisia. Despite the country's limited resources, surveillance of SARS-CoV-2 genomic variation should be continued to control the occurrence of new variants.

Epidemiologic patterns of hepatitis A infection during the pre-vaccination era: A population-based survey in Tunisia in 2015.

BACKGROUND: In the 1980s, Tunisia was considered a country of high endemicity for hepatitis A virus (HAV). Since 2000, an epidemiologic shift has led to an increased incidence of symptomatic and severe forms of HAV infection. OBJECTIVES: In 2015, we conducted a cross-sectional nationwide household-based hepatitis A virus (HAV) seroprevalence study in the total population regardless of age, sex, or geographic origin using a stratified sampling design to make an overview of the HAV epidemiologic situation in Tunisia before vaccine implementation. RESULTS: A total of 6,322 individuals were enrolled. The HAV prevalence was 78.8%. The anti-HAV IgG seropositivity rate increased from 16% for ages 5-9 years to 45% for ages 10-14 years, 67% for ages 15-19 years, 87% for ages 20-24 years, and >90% for older age groups, which suggested an age at midpoint of population immunity (AMPI) in late adolescence. It was significantly higher in rural areas (P < 10-3) and varied significantly between and within regions (P < 10-4). CONCLUSIONS: In this study, although the overall AMPI suggestsintermediate endemicity, the regional AMPI varies from intermediate to very high endemicity profiles attributable to different socioeconomic determinants and conditions of sanitation and hygiene. In addition, it provides insights for the best decisions in terms of vaccination strategies.

[Clinical and epidemiological features of COVID-19-related deaths in Tunisia before the emergence of VOCs (March 2020-February 2021)]. / Caractéristiques cliniques et épidémiologiques des décès COVID-19 en Tunisie avant l'émergence des VOCs (mars 2020-février 2021).

Introduction: the purpose of this study was to describe the clinical and epidemiological features of COVID-19-related deaths in Tunisia notified at the ONMNE (National Observatory of New and emerging Diseases) between 2nd March 2020 and 28th February 2021 and to compare COVID-19-related deaths recorded in Tunisia with the international data. Methods: we conducted a national prospective longitudinal descriptive study of data collected from the National Surveillance System of SARS-CoV-2 infection of the ONMNE, Ministry of Health. All COVID-19-related deaths that occurred in Tunisia between March 2020 and February 2021 were included in this study. Data were collected from hospitals, municipalities and regional health departments. Death notifications were collected from multiple data sources (triangulation): The Regional Directorate of Basic Health Care, the ShocRoom (Strategic Health Operations Center), public and private health facilities, the Crisis Unit of the Presidency of the Government, the Directorate for Hygiene and Environmental Protection, the Ministry of Local Affairs and the Environment, as part of the follow-up of confirmed cases by the ONMNE team, positive RT-PCR / TDR post mortem results. Results: during this study, 8051 deaths were recorded, corresponding to a proportional mortality of 10.4%. The median age was 73 years, with an interquartile range of 17 years. Sex-ratio (M/F) was 1.8. The crude death rate was 69.1/100 000 inhabitants and fatality rate was 3.5%. The analysis of the epidemic curve showed 2 peaks of deaths on 29th October 2020 and 22nd January 2021, with 70 and 86 deaths notified respectively. The spatial distribution of mortality showed that the southern Tunisian region had the highest mortality rate. Patients aged 65 and over were most affected (73.7% of cases) with a crude mortality rate of 570.9/100,000 inhabitants and a fatality rate of 13.7%. Conclusion: prevention strategy based on public health measures must be reinforced by the rapid deployment of anti-COVID-19 vaccination, especially for people at risk of death.

Identification of transmission chains and clusters associated with COVID-19 in Tunisia.

BACKGROUND: The aim of this study was to characterize the transmission chains and clusters of COVID-19 infection in Tunisia. METHODS: All cases were confirmed by Reverse Transcriptase Polymerase Chain Reaction of a nasopharyngeal specimen. Contact tracing is undertaken for all confirmed cases in order to identify close contacts that will be systematically screened and quarantined. Transmission chains were identified based on field investigation, contact tracing, results of screening tests and by assessing all probable mode of transmission and interactions. RESULTS: As of May 18, 2020, 656 cases out of a total of 1043 confirmed cases of Coronavirus disease 2019 belong to 127 transmission chains identified during the epidemic (mean age 42.36 years, Standard deviation 19.56 and sex ratio 0.86). The virus transmission is the most concentrated in the governorate of Tunis (31.5%), Ariana (10.2%) and Ben Arous (10.2%). Virus transmission occurred 50 times (9.72% of secondary transmission events) between two different governorates. A maximum of seven generations of secondary infection was identified, whereas 62% of these secondary infections belong the first generation. A total of 11 "super spreader" cases were identified in this investigation. Four large clusters have been identified. The evolution of secondary cases highlighted two peaks: one in 2nd April and a second in 16 th April whereas imported cases caused local transmission of virus during the early phase of the epidemic. CONCLUSION: Correct contact tracing and early active case finding is useful to identify transmission chains and source of infection in order to contain the widespread transmission in the community.

Characteristics, onset, and evolution of neurological symptoms in patients with COVID-19.

BACKGROUND: A wide range of neurological manifestations has been described in COVID-19. METHODS: In this nationwide retrospective observational study, patients in Tunisia diagnosed with COVID-19 between the 2nd of March and the 16th of May 2020 were contacted by telephone. We collected demographic and clinical data and specified characteristics and evolution of main neurological symptoms. RESULTS: Of 1034 confirmed COVID-19 patients, 646 were included (mean age 42.17 years old) and 466 (72.1%) had neurological symptoms. Neurological symptoms were isolated 22.7% (n = 106). Headache was the most frequent neurological symptom (n = 279, 41.1%): mainly frontotemporal (n = 143, 51.1%) and mild or moderate (n = 165, 59.1%). When associated with fever (n = 143, 51.3%), headache was more likely to be severe and present at onset. Recovery was reported in 83.2%. Smell and taste impairment were found in 37.9% (n = 245) and 36.8% (n = 238) respectively. Among them, 65.3% (156/239) were anosmic and 63.2% (146/231) were ageusic. A complete improvement was found in 72.1% (174/240) of smell impairment and in 76.8% (179/233) of taste impairment. Myalgia (n = 241, 37.3%) and sleep disturbances (n = 241, 37.3%) were also frequent. Imported cases had more neurological symptoms (p = 0.001). In 14.5%, neurological symptoms preceded the respiratory signs (RS). RS were associated with more frequent (p = 0.006) and numerous (p < 0.001) neurological symptoms. CONCLUSIONS: Neurological symptoms in COVID-19 are frequent, can be isolated and present at onset. A total recovery is the most recorded outcome. RS are predictive of neurological symptoms. Studies in to virus and host genetics should be considered to understand the different phenotypes.

Assessment of sample pooling for SARS-CoV-2 molecular testing for screening of asymptomatic persons in Tunisia.

The aim of this study is to test a pooling approach for the RT-PCR test to detect low viral loads of SARS-CoV-2. We found that a single positive specimen can still be detected in pools of up to 10. Each laboratory should conduct its own evaluation and validation of pooling protocols according to its specific context.

Observational study of vaccine effectiveness 20 years after the introduction of universal hepatitis B vaccination in Tunisia.

OBJECTIVES: The objectives of this study were to estimate the national prevalence of hepatitis B infection in Tunisia using data from a nationwide survey, to compare results with those obtained in 1996 survey and to evaluate the impact of vaccination twenty years after its introduction. METHODS: A National household-based cross sectional and serological survey was undertaken in 2015 from randomly selected districts using two-stage sampling. Data collection was performed using standardized and pretested questionnaires and collected blood samples were tested for markers of hepatitis B virus infection. RESULTS: National point prevalence of Hepatitis B surface antigen was 1.7% (95% CI [1.6-1.9%]). The highest prevalence was found in the Center and South regions with respectively 2.3% (95% CI [2.0-2.7%]) and 2.2% (95% CI [1.8-2.8%]). Vaccine effectiveness (VE) was 88.6% (95% CI [81.5-93.0%]) and was higher among population aged less than 20 years 96.1% (95% CI [70.1-99.5%]) than those aged more than 20 years 59.0% (95% CI [32.0-75.3%]). VE was 85.6% (95% CI [65.8-93.9%]) is hyper-endemic areas and 89.1% (95% CI [80.3-94.0%]) in meso-endemic and hypo-endemic areas. CONCLUSIONS: The prevalence of Hepatitis B surface antigen decreased compared to previous estimations and classify Tunisia as a low endemic country as result to the introduction of vaccination since 1995.