A new lineage nomenclature to aid genomic surveillance of dengue virus.

Dengue virus (DENV) is currently causing epidemics of unprecedented scope in endemic settings and expanding to new geographical areas. It is therefore critical to track this virus using genomic surveillance. However, the complex patterns of viral genomic diversity make it challenging to use the existing genotype classification system. Here, we propose adding 2 sub-genotypic levels of virus classification, named major and minor lineages. These lineages have high thresholds for phylogenetic distance and clade size, rendering them stable between phylogenetic studies. We present assignment tools to show that the proposed lineages are useful for regional, national, and subnational discussions of relevant DENV diversity. Moreover, the proposed lineages are robust to classification using partial genome sequences. We provide a standardized neutral descriptor of DENV diversity with which we can identify and track lineages of potential epidemiological and/or clinical importance. Information about our lineage system, including methods to assign lineages to sequence data and propose new lineages, can be found at: dengue-lineages.org.

Socioecological systems analysis of potential factors for cholera outbreaks and assessment of health system's readiness to detect and respond in Ilemela and Nkasi districts, Tanzania.

BACKGROUND: Cholera outbreaks are a recurrent issue in Tanzania, with Ilemela and Nkasi districts being particulary affected. The objective of this study was to conduct a socio-ecological system (SES) analysis of cholera outbreaks in these districts, identifying potential factors and assessing the preparedness for cholera prevention and control. METHODS: A cross-sectional study was carried out in Ilemela and Nkasi districts of Mwanza and Rukwa regions, respectively in Tanzania between September and October 2021. A SES framework analysis was applied to identify potential factors associated with cholera outbreaks and assess the readiness of the districts to cholera prevention and control. RESULTS: Ilemela is characterised by urban and peri-urban ecosystems while Nkasi is mainly rural. Cholera was reported to disproportionately affect people living along the shores of Lake Victoria in Ilemela and Lake Tanganyika in Nkasi, particularly fishermen and women involved infish trading. The main potential factors identified for cholera outbreaks included defecation in the shallow ends and along the edges of lakes, open defecation, bathing/swimming in contaminated waters and improper waste disposal. The preparedness of both districts for cholera prevention and response was found to be inadequate due to limited laboratory capacity, insufficient human resources, and budget constraints. CONCLUSION: People of Ilemela and Nkasi districts remain at significant risk of recurrent cholera outbreaks and the capacity of the districts to detect the disease is limited. Urgent preventive measures, such as conducting considerable community awareness campaigns on personal hygiene and environmental sanitation are needed to alleviate the disease burden and reduce future cholera outbreaks.

Community knowledge, attitude and practices regarding zoonotic viral haemorrhagic fevers in five geo-ecological zones in Tanzania.

BACKGROUND: Viral haemorrhagic fevers (VHF) cause significant economic and public health impact in Sub-Saharan Africa. Community knowledge, awareness and practices regarding such outbreaks play a pivotal role in their management and prevention. This study was carried out to assess community knowledge, attitude and practices regarding VHF in five geo-ecological zones in Tanzania. METHODS: A cross-sectional study was conducted in Buhigwe, Kalambo, Kyela, Kinondoni, Kilindi, Mvomero, Kondoa and Ukerewe districts representing five geo-ecological zones in Tanzania. Study participants were selected by multistage cluster sampling design. A semi-structured questionnaire was used to collect socio-demographic and information related to knowledge, attitude and practices regarding VHFs. Descriptive statistics and logistic regression were used for the analysis. RESULTS: A total of 2,965 individuals were involved in the study. Their mean age was 35 (SD ± 18.9) years. Females accounted for 58.2% while males 41.8%. Most of the respondents (70.6%; n = 2093) had never heard of VHF, and those who heard, over three quarters (79%) mentioned the radio as their primary source of information. Slightly over a quarter (29.4%) of the respondents were knowledgeable, 25% had a positive attitude, and 17.9% had unfavourable practice habits. The level of knowledge varied between occupation and education levels (P < 0.005). Most participants were likely to interact with a VHF survivor or take care of a person suffering from VHF (75%) or visit areas with known VHF (73%). There were increased odds of having poor practice among participants aged 36-45 years (AOR: 3.566, 95% CI: 1.593-7.821) and those living in Western, North-Eastern and Lake Victoria zones (AOR: 2.529, 95% CI: 1.071-6.657; AOR: 2.639, 95% CI: 1.130-7.580 AOR: 2.248, 95% CI: 1.073-3.844, respectively). CONCLUSION: Overall, the knowledge on VHF among communities is low, while a large proportion of individuals in the community are involved in activities that expose them to the disease pathogens in Tanzania. These findings highlight the need for strengthening health educational and promotion efforts on VHF targeting specific populations.

Genomic characterization of Dengue virus circulation in Ethiopia.

In Ethiopia, dengue virus (DENV) infections have been reported in several regions, however, little is known about the circulating genetic diversity. Here, we conducted clinical surveillance for DENV during the 2023 nationwide outbreak and sequenced DENV whole genomes for the first time in Ethiopia. We enrolled patients at three sentinel hospital sites. Using RT-PCR, we screened serum samples for three arboviruses followed by serotyping and sequencing for DENV-positive samples (10.4% of samples). We detected two DENV serotypes (DENV1 and DENV3). Phylogenetic analysis identified one transmission cluster of DENV1 (genotype III major lineage A), and two clusters of DENV3 (genotype III major lineage B). The first showed close evolutionary relationship to the 2023 Italian outbreak and the second cluster to Indian isolates. Co-circulation of DENV1 and DENV3 in some regions of Ethiopia highlights the potential for severe dengue. Intensified surveillance and coordinated public health response are needed to address the threat of severe dengue outbreaks.

A new lineage nomenclature to aid genomic surveillance of dengue virus.

Dengue virus (DENV) is currently causing epidemics of unprecedented scope in endemic settings and expanding to new geographical areas. It is therefore critical to track this virus using genomic surveillance. However, the complex patterns of viral genomic diversity make it challenging to use the existing genotype classification system. Here we propose adding two sub-genotypic levels of virus classification, named major and minor lineages. These lineages have high thresholds for phylogenetic distance and clade size, rendering them stable between phylogenetic studies. We present an assignment tool to show that the proposed lineages are useful for regional, national and sub-national discussions of relevant DENV diversity. Moreover, the proposed lineages are robust to classification using partial genome sequences. We provide a standardized neutral descriptor of DENV diversity with which we can identify and track lineages of potential epidemiological and/or clinical importance. Information about our lineage system, including methods to assign lineages to sequence data and propose new lineages, can be found at: dengue-lineages.org.

Estimating Risk of Introduction of Ebola Virus Disease from the Democratic Republic of Congo to Tanzania: A Qualitative Assessment.

Between April 2018 and November 2020, the Democratic Republic of Congo (DRC) experienced its 11th Ebola virus disease (EVD) outbreak. Tanzania's cross-border interactions with DRC through regular visitors, traders, and refugees are of concern, given the potential for further spread to neighboring countries. This study aimed to estimate the risk of introducing EVD to Tanzania from DRC. National data for flights, boats, and car transport schedules from DRC to Tanzania covering the period of May 2018 to June 2019 were analyzed to describe population movement via land, port, and air travel and coupled with available surveillance data to model the risk of EVD entry. The land border crossing was considered the most frequently used means of travel and the most likely pathway of introducing EVD from DRC to Tanzania. High probabilities of introducing EVD from DRC to Tanzania through the assessed pathways were associated with the viability of the pathogen and low detection capacity at the ports of entry. This study provides important information regarding the elements contributing to the risk associated with the introduction of EBV in Tanzania. It also indicates that infected humans arriving via land are the most likely pathway of EBV entry, and therefore, mitigation strategies including land border surveillance should be strengthened.

Aedes aegypti abundance, larval indices and risk for dengue virus transmission in Kinondoni district, Tanzania.

BACKGROUND: Tanzania has experienced periodic dengue outbreaks with increased incidence since 2010. However, there is limited information on vector dynamics and transmission risk in most parts of the country. This study was conducted to determine Aedes mosquito abundance, larval indices and dengue virus infection rate as risk indicators for DENV transmission in Kinondoni district, Dar es Salaam, Tanzania. METHODS: A cross-sectional study was conducted in three wards of Kinondoni district in Tanzania between December 2019 and January 2020. In each ward, three streets were randomly selected for adult and immature mosquito sampling. The adult mosquitoes were collected using Mosquito Magnet traps, while mosquito larvae and pupae were inspected in water-holding containers in the selected household compounds. The detection of dengue virus (DENV) in female Aedes mosquitoes was done using a one-step reverse transcription-polymerase chain reaction (RT-PCR) method. RESULTS: Of the 1416 adult female mosquitoes collected, Ae. aegypti accounted for 16.8% (n = 238). A total of 333 water-holding containers were inspected and 201 (60.4%) had at least an Aedes larvae or pupae. Water-holding containers supporting the breeding of Aedes larvae and pupae included discarded car tires, flowerpots and small and large plastic containers. The overall House Index, Container Index and Breteau Index were 55.1%, 60.4% and 114.2, respectively. None of the 763 female Aedes mosquitoes tested by RT-PCR was found to be infected with DENV. CONCLUSION: The presence and abundance Ae. aegypti mosquitoes and the large proportion of water-holding containers infested with the mosquito larvae and pupae put residents of Kinondoni district at high risk of DENV transmission. Our findings emphasize the need for continuous mosquito vector surveillance and control to prevent the possibility of future DENV outbreaks in Tanzania.

Viral haemorrhagic fevers and malaria co-infections among febrile patients seeking health care in Tanzania.

BACKGROUND: In recent years there have been reports of viral haemorrhagic fever (VHF) epidemics in sub-Saharan Africa where malaria is endemic. VHF and malaria have overlapping clinical presentations making differential diagnosis a challenge. The objective of this study was to determine the prevalence of selected zoonotic VHFs and malaria co-infections among febrile patients seeking health care in Tanzania. METHODS: This facility-based cross-sectional study was carried out between June and November 2018 in Buhigwe, Kalambo, Kyela, Kilindi, Kinondoni, Kondoa, Mvomero, and Ukerewe districts in Tanzania. The study involved febrile patients seeking health care from primary healthcare facilities. Blood samples were collected and tested for infections due to malaria, Crimean-Congo haemorrhagic fever (CCHF), Ebola virus disease (EVD), Marburg virus disease (MVD), Rift Valley fever (RVF) and yellow fever (YF). Malaria infections were tested using rapid diagnostics tests while exposure to VHFs was determined by screening for immunoglobulin M antibodies using commercial enzyme-linked immunosorbent assays. The Chi-square test was used to compare the proportions. RESULTS: A total of 308 participants (mean age = 35 ± 19 years) were involved in the study. Of these, 54 (17.5%) had malaria infection and 15 (4.8%) were positive for IgM antibodies against VHFs (RVF = 8; CCHF = 2; EBV = 3; MBV = 1; YF = 1). Six (1.9%) individuals had both VHF (RVF = 2; CCHF = 1; EVD = 2; MVD = 1) and malaria infections. The highest co-infection prevalence (0.6%) was observed among individuals aged 46‒60 years (P < 0.05). District was significantly associated with co-infection (P < 0.05) with the highest prevalence recorded in Buhigwe (1.2%) followed by Kinondoni (0.9%) districts. Headache (100%) and muscle, bone, back and joint pains (83.3%) were the most significant complaints among those infected with both VHFs and malaria (P = 0.001). CONCLUSIONS: Co-infections of VHF and malaria are prevalent in Tanzania and affect more the older than the younger population. Since the overlapping symptoms in co-infected individuals may challenge accurate diagnosis, adequate laboratory diagnosis should be emphasized in the management of febrile illnesses.

Circulation of dengue serotype 1 viruses during the 2019 outbreak in Dar es Salaam, Tanzania.

Dengue is an important mosquito-borne viral disease in humans in tropical and subtropical countries. In 2019, a total of 6917 dengue cases were reported in Tanzania based on serological analysis. The aim of this study was to confirm the presence of dengue virus (DENV) and conduct its genetic characterization. A total of 191 serum samples were collected from the outpatients seeking care from health facilities in Kinondoni and Ilala districts between March and May 2019. All the samples were initially tested for the presence of non-structural protein 1 and anti-DENV immunoglobulin G (IgG) and IgM using a commercial OnSite Duo Dengue Ag-IgG/IgM rapid test. Of the 191 sera, 110 (57.6%) were DENV seropositive. The presence of DENV ribonucleic acid was confirmed in 18.2% of the seropositive sera by reverse transcription polymerase chain reaction (RT-PCR). The RT-PCR products were cleaned and partial sequences of DENV polyprotein gene determined using dideoxynucleotide cycle sequencing followed by phylogenetic analysis. We present the occurrence of DENV serotype 1 (DENV-1) during the 2019 outbreak in Tanzania. The DENV-1 strains reported in the present study are highly identical and cluster with Asian DENV-1 strains indicating the possibility of intercontinental spread of DENV through globalization. We advocate for the need for molecular surveillance of dengue viruses during outbreaks to provide rapid evidence of the disease to guide public health interventions.

Seroprevalence and associated risk factors of selected zoonotic viral hemorrhagic fevers in Tanzania.

OBJECTIVE: To determine the seroprevalence of selected zoonotic viral hemorrhagic fevers (VHFs) and their associated risk factors in Tanzania. METHODS: Blood samples were collected from consenting outpatients and community members in eight districts selected from five ecological zones of Tanzania. Serum was harvested and tested for the presence of immunoglobulin G (IgG) and M (IgM) antibodies against Crimean-Congo hemorrhagic fever (CCHF), Ebola virus disease (EVD), Marburg virus disease (MVD), Rift Valley fever (RVF), and yellow fever (YF). RESULTS: The presence of IgM and IgG antibodies against CCHF, EVD, MVD, RVF, and YF was detected in 64 of 500 samples (12.8%). The prevalences of IgM and IgG antibodies to CCHF, EVD, MVD, RFV, and YF were 2.0%, 3.4%, 1.2%, 4.8%, and 1.4%, respectively. Contact with wild animals (OR = 1.2, CI = 1.3-1.6) and keeping goats (OR = 1.3, CI = 1.5-1.9) were significantly associated with RVF, while contact with bats (OR = 1.2, CI = 1.1-1.5) was associated with MVD. CONCLUSION: The findings of this study provide evidence of exposure to CCHF, EVD, MVD, RVF, and YF in Tanzania. Since most of these VHFs occurred without apparent clinical forms of the disease, these findings call for the need to strengthen the surveillance system and management of febrile illnesses in Tanzania.

Dengue Virus Infection and Associated Risk Factors in Africa: A Systematic Review and Meta-Analysis.

Dengue contributes a significant burden on global public health and economies. In Africa, the burden of dengue virus (DENV) infection is not well described. This review was undertaken to determine the prevalence of dengue and associated risk factors. A literature search was done on PubMed/MEDLINE, Scopus, Embase, and Google Scholar databases to identify articles published between 1960 and 2020. Meta-analysis was performed using a random-effect model at a 95% confidence interval, followed by subgroup meta-analysis to determine the overall prevalence. Between 1960 and 2020, 45 outbreaks were identified, of which 17 and 16 occurred in East and West Africa, respectively. Dengue virus serotype 1 (DENV-1) and DENV-2 were the dominant serotypes contributing to 60% of the epidemics. Of 2211 cases reported between 2009 and 2020; 1954 (88.4%) were reported during outbreaks. Overall, the prevalence of dengue was 29% (95% CI: 20-39%) and 3% (95% CI: 1-5%) during the outbreak and non-outbreak periods, respectively. Old age (6/21 studies), lack of mosquito control (6/21), urban residence (4/21), climate change (3/21), and recent history of travel (3/21) were the leading risk factors. This review reports a high burden of dengue and increased risk of severe disease in Africa. Our findings provide useful information for clinical practice and health policy decisions to implement effective interventions.

Seroprevalence and associated risk factors of chikungunya, dengue, and Zika in eight districts in Tanzania.

BACKGROUND: This study was conducted to determine the seroprevalence and risk factors of chikungunya (CHIKV), dengue (DENV), and Zika (ZIKV) viruses in Tanzania. METHODS: The study covered the districts of Buhigwe, Kalambo, Kilindi, Kinondoni, Kondoa, Kyela, Mvomero, and Ukerewe in Tanzania. Blood samples were collected from individuals recruited from households and healthcare facilities. An ELISA was used to screen for immunoglobulin G antibodies against CHIKV, DENV, and ZIKV. RESULTS: A total of 1818 participants (median age 34 years) were recruited. The overall CHIKV, DENV, and ZIKV seroprevalence rates were 28.0%, 16.1%, and 6.8%, respectively. CHIKV prevalence was highest in Buhigwe (46.8%), DENV in Kinondoni (43.8%), and ZIKV in Ukerewe (10.6%) and Mvomero (10.6%). Increasing age and frequent mosquito bites were significantly associated with CHIKV and DENV seropositivity (P < 0.05). Having piped water or the presence of stagnant water around the home (P < 0.01) were associated with higher odds of DENV seropositivity. Fever was significantly associated with increased odds of CHIKV seropositivity (P < 0.001). Visiting mines had higher odds of ZIKV seropositivity (P < 0.05). CONCLUSIONS: These findings indicate that DENV, CHIKV, and ZIKV are circulating in diverse ecological zones of Tanzania. There is a need to strengthen the control of mosquito-borne viral diseases in Tanzania.

Forty-two years of responding to Ebola virus outbreaks in Sub-Saharan Africa: a review.

Introduction: Ebola virus disease (EVD) is one of the deadliest haemorrhagic fevers affecting humans and non-human primates. Thirty-four outbreaks have been reported in Africa since it was first recognised in 1976. This review analysed 42 years of EVD outbreaks and identified various challenges and opportunities for its control and prevention in Sub-Saharan Africa. Methods: A literature search of relevant articles on EVD was done in PubMed, Web of Science and Google Scholar electronic databases. Articles published from 1976 to 2019 were reviewed to document reports of EVD outbreaks in Sub-Saharan Africa. Data extraction focused on the year of outbreak, geographical spread, virus strain involved, number of cases and deaths, case fatality, and outbreak management. Analyses of trends in case fatality were performed by calculating ORs between times. Results: In the past four decades, a total of 34 EVD outbreaks affecting 34 356 cases and causing 14 823 deaths were reported in 11 countries in Sub-Saharan Africa. The overall case fatality rate (95% CI) was 66% (62 to 71) and did not change substantially over time (OR in 2019 vs 1976=1.6 (95% CI 1.5 to 1.8), p<0.001). The results of this review indicate that challenges to control EVD outbreaks are related to epidemiological, sociocultural and health system factors. Conclusions: Sub-Saharan Africa continues to face considerable challenges in EVD control, whereby there are no significant changes in case fatality rates observed during the past four decades. Socioeconomic and cultural processes need to be critically considered to shape the community behaviours that lead to exposure to EVD outbreaks. Areas that need to be addressed to prevent future EVD outbreaks include a broad-based, one-health approach, effective communication, social mobilisation, and strengthening of the health systems.

Load and Prevalence of Antimicrobial-Resistant Escherichia coli from Fresh Goat Meat in Arusha, Tanzania.

Given the potential public health risks associated with a burgeoning goat meat industry in Tanzania, we estimated the load of Escherichia coli and the prevalence of antibiotic-resistant strains for goat meat by using a cross-sectional study design (June to July 2015). Five large (n = 60 samples) and five small (n = 64 samples) slaughterhouses were sampled over a period of four to six visits each. Meat rinsate was prepared and plated onto MacConkey agar, and presumptive E. coli colonies were enumerated and reported as CFU per milliliter of rinsate. In total, 2,736 presumptive E. coli isolates were tested for antibiotic drug sensitivity by using breakpoint assays against 11 medically important antibiotics. E. coli was recovered from almost all the samples (96.8%), with counts ranging from 2 to 4 log CFU ml-1, and there was no significant difference (P = 0.43) in recovery according to facility size (average, 3.37 versus 3.13 log CFU ml-1, large and small, respectively). Samples from large facilities had relatively higher prevalence (P = 0.026) of antibiotic-resistant E. coli compared with small facilities. This was mostly explained by more ampicillin (30.1 versus 12.8%) and amoxicillin (17.6 versus 4.5%) resistance for large versus small facilities, respectively, and more tetracycline resistance for small facilities (5.6 versus 10.6%, respectively). Large slaughter operations may serve as foci for dissemination of antibiotic-resistant bacteria via food products. More effective hygiene practices during slaughter and meat handling would limit the probability of transmitting antibiotic-resistant E. coli in goat meat.

Prevalence of Antibiotic-Resistant Fecal Escherichia coli Isolates from Penned Broiler and Scavenging Local Chickens in Arusha, Tanzania.

We compared the prevalence of antibiotic-resistant Escherichia coli isolates from household-level producers of broiler (commercial source breeds) and local chickens in the Arusha District of Tanzania. Households were composed of a single dwelling or residence with independent, penned broiler flocks. Free-range, scavenging chickens were mixed breed and loosely associated with individual households. A total of 1,800 E. coli isolates (1,200 from broiler and 600 from scavenging local chickens) from 75 chickens were tested for their susceptibility against 11 antibiotics by using breakpoint assays. Isolates from broiler chickens harbored a higher prevalence of antibiotic-resistant E. coli relative to scavenging local chickens, including sulfamethoxazole (80.3 versus 34%), followed by trimethoprim (69.3 versus 27.7%), tetracycline (56.8 versus 20%), streptomycin (52.7 versus 24.7%), amoxicillin (49.6 versus 17%), ampicillin (49.1 versus 16.8%), ciprofloxacin (21.9 versus 1.7%), and chloramphenicol (1.5 versus 1.2%). Except for resistance to chloramphenicol, scavenging local chickens harbored fewer resistant E. coli isolates (P < 0.05). Broiler chickens harbored more isolates that were resistant to ≥7 antibiotics (P < 0.05). The higher prevalence of antibiotic-resistant E. coli from broiler chickens correlated with the reported therapeutic and prophylactic use of antibiotics in this poultry population. We suggest that improved biosecurity measures and increased vaccination efforts would reduce reliance on antibiotics by these households.