Cholera and diarrheal diseases in Cuamba District, Niassa Province, Mozambique: Systematic healthcare facility-based surveillance strengthening, characteristics of suspected cholera and diarrheal patients, and incidence of diarrheal diseases.

BACKGROUND: Mozambique is one of the countries in Africa that is continuously at risk of cholera outbreaks due to poor sanitation, hygiene, and limited access to potable water in some districts. The Mozambique Cholera Prevention and Surveillance (MOCA) project was implemented in Cuamba District, Niassa Province to prevent and control cholera outbreaks through a preemptive cholera vaccination, strengthened surveillance system for cholera and diarrheal diseases, and better understanding of cholera-related healthcare seeking behavior of local populations, which may further guide the national cholera control and prevention strategies. This article presents the surveillance component of the MOCA project. METHODOLOGY/PRINCIPAL FINDINGS: A prospective healthcare facility (HCF)-based surveillance of cholera and diarrheal disease was conducted in six HCFs in the District of Cuamba from March 2019 to December 2020. A systematic surveillance procedure has been put in place with capacity building in selected sentinel HCFs and a basic microbiology laboratory established on-site. Patients presenting with suspected cholera or other diarrheal symptoms were eligible for enrollment. Clinical data and rectal swab samples were collected for laboratory confirmation of Vibrio Cholerae and other pathogens. A total of 419 eligible patients from six HCFs were enrolled. The median age was 19.8 years with a similar age distribution between sentinel sites. The majority were patients who exhibited diarrhea symptoms not suspected of cholera (88.8%; n = 410). Among those, 59.2% (210/397) were female and 59.9% (235/392) were 15 years and above. There were 2 cholera cases, coming outside of the catchment area. The incidence of diarrheal diseases ranged from 40-103 per 100,000 population. No Vibrio cholerae was isolated among surveillance catchment population and Escherichia coli spp. (82/277; 29.6%) was the most common pathogen isolated. CONCLUSION/SIGNIFICANCE: Efforts were made to strengthen the systematic surveillance of suspected cholera with standardised patient screening, enrolment, and diagnostics. The first basic microbiology laboratory in Niassa Province established in Cuamba District under the MOCA project needs to be integrated into the national network of laboratories for sustainability. No reports of laboratory confirmed cholera cases from the surveillance catchment area may be highly related to the pre-emptive oral cholera vaccine (OCV) mass vaccination campaign conducted in 2018 and the use of drugs by local populations prior to visiting the sentinel HCFs. Continued systematic cholera surveillance is needed to closely monitor the cholera endemicity and epidemics, and further evaluate the long-term impact of this vaccination. High incidence of diarrheal illnesses needs to be addressed with improved water, sanitation, and hygiene (WaSH) conditions in Cuamba District. Efforts integrated with the prioritization of prevention measures are fundamental for the control of cholera in the country.

Pre-emptive oral cholera vaccine (OCV) mass vaccination campaign in Cuamba District, Niassa Province, Mozambique: feasibility, vaccination coverage and delivery costs using CholTool.

INTRODUCTION: Mozambique suffers from regular floods along its principal river basins and periodic cyclones that resulted in several cholera epidemics during the last decades. Cholera outbreaks in the recent 5 years affected particularly the northern provinces of the country including Nampula and Niassa provinces. A pre-emptive oral cholera vaccine (OCV) mass vaccination campaign was conducted in Cuamba District, Niassa Province, and the feasibility, costs, and vaccination coverage assessed. METHODS: WHO prequalified OCV (Euvichol-Plus), a killed whole-cell bivalent vaccine containing Vibrio cholerae O1 (classical and El Tor) and O139, was administered in two doses with a 15-day interval during 7-31 August 2018, targeting around 180 000 people aged above 1 year in Cuamba District. Microplanning, community sensitisation, and training of local public health professionals and field enumerators were conducted. Feasibility and costs of vaccination were assessed using CholTool. Vaccination coverage and barriers were assessed through community surveys. RESULTS: The administrative coverage of the first and second rounds of the campaign were 98.9% (194 581) and 98.8% (194 325), respectively, based on the available population data that estimated total 196 652 inhabitants in the target area. The vaccination coverage survey exhibited 75.9% (±2.2%) and 68.5% (±3.3%) coverage for the first and second rounds, respectively. Overall, 60.4% (±3.4%) of the target population received full two doses of OCV. Barriers to vaccination included incompatibility between working hours and campaign time. No severe adverse events were notified. The total financial cost per dose delivered was US$0.60 without vaccine cost and US$1.98 including vaccine costs. CONCLUSION: The pre-emptive OCV mass vaccination campaign in remote setting in Mozambique was feasible with reasonable full-dose vaccination coverage to confer sufficient herd immunity for at least the next 3 to 5 years. The delivery cost estimate indicates that the OCV campaign is affordable as it is comparable with Gavi's operational support for vaccination campaigns.

Mapeamento de áreas críticas de cólera em moçambique / Mapping critical cholera areas in Mozambique

Moçambique é signatário da iniciativa global para a eliminação da cólera até ao ano de 2030, estratégia voltada para a interrupção da transmissão da doença, redução da mortalidade e eliminação da cólera como problema de saúde pública. Em Moçambique, a cólera é altamente sazonal, com acentuada concentração de casos durante o período quente e chuvoso, especialmente nas províncias de Cabo Delgado, Nampula, Tete e Sofala. É neste contexto, que se conduziu a análise de situação da cólera com o objectivo de identificar e mapear todas as zonas quentes e de alto risco para a doença em todo território nacional. E através de uma análise epidemiológica multinível da cólera e da Diarréia aguda, em todos os distritos e Postos administrativos do país, com recurso a dados retrospetivos dos anos 2017 à 2021, foram identificadas as zonas quentes e de alto risco da cólera em Moçambique de que se faz a presente descrição. Os métodos para mapeamento de zonas quentes foram concebidos a partir da orientação do GTFCC para identificação de zonas quentes de cólera, revistos e adaptados ao contexto nacional. Os dados utilizados foram dos casos de cólera, casos, internamentos e óbitos por Diarréia aguda notificados a nível distrital, registados na base de dados nacional de saúde (SIS-MA) e reportados pelos sistemas de gestão de dados da vigilância epidemiológica das Direcções Provinciais de Saúde (DPS's). Uma "abordagem experimental" para inferir a ocorrência de cólera a partir da ocorrência de doença diarreica foi realizada, através de dados de doença grave e óbitos por Diarréia aguda em adultos. Uma ferramenta Excel foi desenvolvida para facilitar a entrada e análise de dados por província. Todos os distritos do país foram classificados de acordo com a carga de cólera ou doença diarreica aguda e os com elevada pontuação foram selecionados e seus postos administrativos (PA) foram mapeados. Indicadores epidemiológicos adicionais e factores de risco foram utilizados para afinar e qualificar a selecção dos PA zonas quentes ou de alto risco da cólera. Foram identificados 250 Postos administrativos críticos para intervenção no contexto da eliminação da cólera, destes 75 são zonas quentes e 175 são zonas de alto risco com uma estimativa global de 14 587 782 habitantes afectados. Neste contexto, recomenda-se conceber, validar e implementar um plano nacional de eliminação da cólera com os subplanos específicos para cada pilar anexados e estabelecer um programa nacional de eliminação da cólera.

Conditions to eliminate cholera in Mozambique - the pathway for the development of the national cholera plan.

Cholera disproportionately affects the most vulnerable segments of the population, particularly those who have low or no access to basic water, sanitation, and hygiene (WASH). Despite some improvements in WASH conditions, cholera still represents a persistent challenge in Mozambique, where outbreaks occur almost every year, with high case fatality rates, posing a threat to the country's economic development. The Government of Mozambique has started developing a revised National Cholera Plan (NCP), which aligns with "ending cholera-a global roadmap to 2030" launched by the Global Task Force on Cholera Control (GTFCC) in 2017. Ending cholera represents a critical step towards achieving the sustainable development goals and requires effective prevention and control interventions, ensuring that no one is left behind. The NCP must use a multi-sector approach and broad stakeholder collaboration with well-coordinated roles and functions of different partners to address major areas for cholera elimination - water and sanitation, health care services and management, epidemiology and surveillance, and health and hygiene promotion. Every cholera death is preventable. In this review, we reiterate the need for effective coordinated actions to control and eliminate cholera in Mozambique and decrease the cholera burden, enabling a healthy population over the generations.

Microbiological contamination of improved water sources, Mozambique.

Objective: To assess if water from improved sources are microbiologically safe in Niassa province, Mozambique, by examining the presence of total coliforms in different types of water sources. Methods: We conducted a cross-sectional household survey in two rural districts of Niassa province during the dry season, from 21 August to 4 October 2019. We observed water sources and conducted microbiological water quality tests and structured household interviews. Findings: We included 1313 households, of which 812 (61.8%) used water from an improved source. There was no significant difference in presence of total coliforms between water sampled at improved and unimproved water sources, 62.7% (509 samples) and 65.7% (329 samples), respectively (P-value = 0.267). Households using improved water sources spent significantly longer time collecting water (59.1 minutes; standard deviation, SD: 55.2) than households using unimproved sources (49.8 minutes; SD: 58.0; P-value < 0.001). A smaller proportion of households using improved sources had access to water sources available 24 hours per day than that of households using unimproved sources, 71.7% (582 households) versus 94.2% (472 households; P-value < 0.001). Of the 240 households treating water collected from improved sources, 204 (85.4%) had total coliforms in their water, while treated water from 77 of 107 (72.0%) households collecting water from an unimproved source were contaminated. Conclusion: Current access to an improved water source does not ensure microbiological safety of water and thereby using access as the proxy indicator for safe drinking and cooking water is questionable. Poor quality of water calls for the need for integration of water quality assessment into regular monitoring programmes.

Molecular characterization of diarrheagenic Escherichia coli isolates from children with diarrhea: A cross-sectional study in four provinces of Mozambique: Diarrheagenic Escherichia coli in Mozambique.

OBJECTIVES: Analyze the frequency of diarrheagenic Escherichia coli (DEC) pathotypes and their antimicrobial resistance profiles among children aged <15 years with diarrhea in four Mozambican provinces. METHODS: A cross-sectional hospital-based surveillance program of diarrhea was implemented in Maputo, Sofala, Zambézia, and Nampula. A single stool sample was collected from each child from May 2014 to May 2017. Culture methods and biochemical characterization were performed to detect E. coli strains. DEC pathotypes were determined by conventional polymerase chain reaction targeting specific virulence genes. Antimicrobial susceptibility was assessed by the Kirby-Bauer method. RESULTS: From 723 specimens analyzed by culture, 262 were positive for E. coli. A total of 208 samples were tested by polymerase chain reaction for DEC identification, of which 101 (48.6%) were positive for a DEC pathotype. The predominant pathotypes were enteroaggregative (66.3%, 67/101), enteropathogenic (15.8%, 16/101), enterotoxigenic (13.9%, 14/101), and enteroinvasive E. coli (4.0%, 4/101). No Shiga toxin-producing E. coli was identified. Regardless of the province, the most frequent pathotype was enteroaggregative E. coli. Isolated DEC presented high frequency of resistance to ampicillin (97.8%), tetracycline (68.3%), chloramphenicol (28.4%), nalidixic acid (19.5%), and gentamicin (14.4%). CONCLUSION: Children with diarrhea in Mozambique had DEC and higher resistance to ampicillin and tetracycline.

The seasonality of cholera in sub-Saharan Africa: a statistical modelling study.

BACKGROUND: Cholera remains a major threat in sub-Saharan Africa (SSA), where some of the highest case-fatality rates are reported. Knowing in what months and where cholera tends to occur across the continent could aid in improving efforts to eliminate cholera as a public health concern. However, largely due to the absence of unified large-scale datasets, no continent-wide estimates exist. In this study, we aimed to estimate cholera seasonality across SSA and explore the correlation between hydroclimatic variables and cholera seasonality. METHODS: Using the global cholera database of the Global Task Force on Cholera Control, we developed statistical models to synthesise data across spatial and temporal scales to infer the seasonality of excess (defined as incidence higher than the 2010-16 mean incidence rate) suspected cholera occurrence in SSA. We developed a Bayesian statistical model to infer the monthly risk of excess cholera at the first and second administrative levels. Seasonality patterns were then grouped into spatial clusters. Finally, we studied the association between seasonality estimates and hydroclimatic variables (mean monthly fraction of area flooded, mean monthly air temperature, and cumulative monthly precipitation). FINDINGS: 24 (71%) of the 34 countries studied had seasonal patterns of excess cholera risk, corresponding to approximately 86% of the SSA population. 12 (50%) of these 24 countries also had subnational differences in seasonality patterns, with strong differences in seasonality strength between regions. Seasonality patterns clustered into two macroregions (west Africa and the Sahel vs eastern and southern Africa), which were composed of subregional clusters with varying degrees of seasonality. Exploratory association analysis found most consistent and positive correlations between cholera seasonality and precipitation and, to a lesser extent, between cholera seasonality and temperature and flooding. INTERPRETATION: Widespread cholera seasonality in SSA offers opportunities for intervention planning. Further studies are needed to study the association between cholera and climate. FUNDING: US National Aeronautics and Space Administration Applied Sciences Program and the Bill & Melinda Gates Foundation.

Immunogenicity of Reduced-Dose Monovalent Type 2 Oral Poliovirus Vaccine in Mocuba, Mozambique.

BACKGROUND: The monovalent type 2 oral poliovirus vaccine (mOPV2) stockpile is low. One potential strategy to stretch the existing mOPV2 supply is to administer a reduced dose: 1 drop instead of 2. METHODS: We conducted a randomized, controlled, open-label, noninferiority trial (10% margin) to compared immunogenicity after administration of 1 versus 2 drops of mOPV2. We enrolled 9-22-month-old infants from Mocuba district of Mozambique. Poliovirus neutralizing antibodies were measured in serum samples collected before and 1 month after mOPV2 administration. Immune response was defined as seroconversion from seronegative (<1:8) at baseline to seropositive (≥1:8) after vaccination or boosting titers by ≥4-fold for those with titers between 1:8 and 1:362 at baseline. The trial was registered at anzctr.org.au (no. ACTRN12619000184178p). RESULTS: We enrolled 378 children, and 262 (69%) completed per-protocol requirements. The immune response of mOPV2 was 53.6% (95% confidence interval, 44.9%-62.1%) and 60.6% (52.2%-68.4%) in 1-drop and 2-drop recipients, respectively. The noninferiority margin of the 10% was not reached (difference, 7.0%; 95% confidence interval, -5.0% to 19.0%). CONCLUSION: A small loss of immunogenicity of reduced mOPV2 was observed. Although the noninferiority target was not achieved, the Strategic Advisory Group of Experts on Immunization recommended the 1-drop strategy as a dose-sparing measure if mOPV2 supplies deteriorate further.

Immunogenicity of reduced-dose monovalent Type 2 oral poliovirus vaccine in mocuba, mozambique

The monovalent type 2 oral poliovirus vaccine (mOPV2) stockpile is low. One potential strategy to stretch the existing mOPV2 supply is to administer a reduced dose: 1 drop instead of 2.

Spectrum of nutrition-specific and nutrition-sensitive determinants of child undernutrition: a multisectoral cross-sectional study in rural Mozambique.

BACKGROUND: Despite an increasing need for multisectoral interventions and coordinations for addressing malnutrition, evidence-based multisectoral nutrition interventions have been rarely developed and implemented in low-income and middle-income countries. To identify key determinants of undernutrition for effectively designing a multisectoral intervention package, a nutrition survey was conducted, by comprehensively covering a variety of variables across sectors, in Niassa province, Mozambique. METHODS: A cross-sectional household survey was conducted in Niassa province, August-October 2019. Anthropometric measurements, anaemia tests of children under 5 years of age and structured interviews with their mothers were conducted. A total of 1498 children under 5 years of age participated in the survey. We employed 107 background variables related to possible underlying and immediate causes of undernutrition, to examine their associations with being malnourished. Both bivariate (χ2 test and Mann-Whitney's U test) and multivariate analyses (logistic regression) were undertaken, to identify the determinants of being malnourished. RESULTS: Prevalence rates of stunting, underweight and wasting were estimated at 46.2%, 20.0% and 7.1%, respectively. Timely introduction of solid, semi-solid or soft foods to children of 6-8 months of age was detected as a determinant of being not stunted. Mother-child cosleeping and ownership of birth certificate were a protective factor from and a promoting factor for being underweight, respectively. Similarly, availability and consumption of eggs at the household level and cough during the last 2 weeks among children were likely to be a protective factor from and a promoting factor for being wasted, respectively. CONCLUSION: Timely introduction of solid, semi-solid or soft foods could serve as an entry point for the three sectors to start making joint efforts, as it requires the interventions from all health, agriculture and water sectors. To enable us to make meaningful interprovincial, international and inter-seasonal comparisons, it is crucially important to develop a standard set of variables related to being malnourished.

Epidemiological profile of health consultations during the Mozambique 9th National Cultural Festival, August 2016.

INTRODUCTION: The use of mass gatherings as spaces to practice health surveillance has been growing in recent years. In Mozambique, the 9th National Festival of Culture in 2016 was selected for this practice. A specific public health surveillance system to facilitate rapid detection of outbreaks and other health-related events was implemented for this event with real time data collection and analysis. METHODS: A descriptive epidemiological evaluation of all the health consultations that occurred in fixed posts prepared for the event was conducted. The data were collected through electronic mobile system (tablets) in real time, with the aid of a form designed for this purpose and sent directly to the incident command system (ICS). RESULTS: During the event, a total of 355 patients were assisted, 52.3% were female, 87.0% were from Beira city and the artists were the group that most frequently sought health care at 59.4%. The largest number of visits took place on the third day (36.4%). People over 45 years of age were the age group that most frequently sought health care (30.8%). The main provisional diagnoses of those who were attended to during the festival was arterial hypertension (20.3%), followed by febrile syndrome (19.0%), with falls being the most frequent causes of trauma during the festival (60.0%). CONCLUSION: The system of monitoring in real time using mobile technologies proved to be efficient for the monitoring of the main health events during the mass gatherings. This profile of health consultations encourages the health sector to plan strategies and actions geared to the reality of care for this type of event.

Oral cholera vaccine coverage during a preventive door-to-door mass vaccination campaign in Nampula, Mozambique.

BACKGROUND: In addition to improving water, sanitation and hygiene (WASH) measures and optimal case management, the introduction of Oral cholera vaccine (OCV) is a complementary strategy for cholera prevention and control for vulnerable population groups. In October 2016, the Mozambique Ministry of Health implemented a mass vaccination campaign using a two-dose regimen of the Shanchol™ OCV in six high-risk neighborhoods of Nampula city, in Northern Mozambique. Overall 193,403 people were targeted by the campaign, which used a door-to-door strategy. During campaign follow-up, a population survey was conducted to assess: (1) OCV coverage; (2) frequency of adverse events following immunization; (3) vaccine acceptability and (4) reasons for non-vaccination. METHODOLOGY/PRINCIPAL FINDINGS: In the absence of a household listing and clear administrative neighborhood delimitations, we used geospatial technology to select households from satellite images and used the support of community leaders. One person per household was randomly selected for interview. In total, 636 individuals were enrolled in the survey. The overall vaccination coverage with at least one dose (including card and oral reporting) was 69.5% (95%CI: 51.2-88.2) and the two-dose coverage was 51.2% (95%CI: 37.9-64.3). The campaign was well accepted. Among the 185 non-vaccinated individuals, 83 (44.6%) did not take the vaccine because they were absent when the vaccination team visited their houses. Among the 451 vaccinated individuals, 47 (10%) reported minor and non-specific complaints, and 78 (17.3%) mentioned they did not receive any information before the campaign. CONCLUSIONS/SIGNIFICANCE: In spite of overall coverage being slightly lower than expected, the use of a mobile door-to-door strategy remains a viable option even in densely-populated urban settings. Our results suggest that campaigns can be successfully implemented and well accepted in Mozambique in non-emergency contexts in order to prevent cholera outbreaks. These findings are encouraging and complement the previous Mozambican experience related to OCV.

Multi-site cholera surveillance within the African Cholera Surveillance Network shows endemicity in Mozambique, 2011-2015.

BACKGROUND: Mozambique suffers recurrent annual cholera outbreaks especially during the rainy season between October to March. The African Cholera Surveillance Network (Africhol) was implemented in Mozambique in 2011 to generate accurate detailed surveillance data to support appropriate interventions for cholera control and prevention in the country. METHODOLOGY/PRINCIPAL FINDINGS: Africhol was implemented in enhanced surveillance zones located in the provinces of Sofala (Beira), Zambézia (District Mocuba), and Cabo Delgado (Pemba City). Data were also analyzed from the three outbreak areas that experienced the greatest number of cases during the time period under observation (in the districts of Cuamba, Montepuez, and Nampula). Rectal swabs were collected from suspected cases for identification of Vibrio cholerae, as well as clinical, behavioral, and socio-demographic variables. We analyzed factors associated with confirmed, hospitalized, and fatal cholera using multivariate logistic regression models. A total of 1,863 suspected cases and 23 deaths (case fatality ratio (CFR), 1.2%) were reported from October 2011 to December 2015. Among these suspected cases, 52.2% were tested of which 23.5% were positive for Vibrio cholerae O1 Ogawa. Risk factors independently associated with the occurrence of confirmed cholera were living in Nampula city district, the year 2014, human immunodeficiency virus infection, and the primary water source for drinking. CONCLUSIONS/SIGNIFICANCE: Cholera was endemic in Mozambique during the study period with a high CFR and identifiable risk factors. The study reinforces the importance of continued cholera surveillance, including a strong laboratory component. The results enhanced our understanding of the need to target priority areas and at-risk populations for interventions including oral cholera vaccine (OCV) use, and assess the impact of prevention and control strategies. Our data were instrumental in informing integrated prevention and control efforts during major cholera outbreaks in recent years.

Antibiotics resistance in El Tor Vibrio cholerae 01 isolated during cholera outbreaks in Mozambique from 2012 to 2015.

RATIONALE: Mozambique has recorded cyclically epidemic outbreaks of cholera. Antibiotic therapy is recommended in specific situations for management and control of cholera outbreaks. However, an increase in resistance rates to antibiotics by Vibrio cholerae has been reported in several epidemic outbreaks worldwide. On the other hand, there are few recent records of continuous surveillance of antibiotics susceptibility pattern of V. cholerae in Mozambique. GOALS: The purpose of this study was to evaluate antibiotics resistance pattern of Vibrio cholerae O1 Ogawa isolated during Cholera outbreaks in Mozambique to commonly used antibiotics. METHODOLOGY: We analyzed data from samples received in the context of surveillance and response to Cholera outbreaks in the National Reference Laboratory of Microbiology from the National Institute of Health of Mozambique, 159 samples suspected of cholera from cholera treatment centers of, Metangula (09), Memba (01), Tete City (08), Moatize (01), Morrumbala (01) districts, City of Quelimane (01), Lichinga (06) and Nampula (86) districts, from 2012 to 2015. Laboratory culture and standard biochemical tests were employed to isolate and identify Vibrio cholerae; serotypes were determined by antisera agglutination reaction in blade. Biotype and presence of important virulence factors analysis was done by PCR. Antibiotics susceptibility pattern was detected by disk diffusion method Kirby Bauer. Antibiotic susceptibility and results were interpreted by following as per recommendations of CLSI (Clinical and Laboratory Standards Institute) 2014. All samples were collected and tested in the context of Africhol Project, approved by the National Bioethics Committee for Health. RESULTS: Among isolates from of Vibrio cholerae O1 El Tor Ogawa resistance to Sulphamethoxazole-trimethropim was 100% (53/53) to Trimethoprim-, being 100% (54/54) for Ampicillin, 99% (72/74) for Nalidixic Acid, 97% (64/66) to Chloramphenicol, 95% (42/44) for Nitrofurantoin and (19/20) Cotrimoxazole, 83% (80/97) Tetracycline, 56% (5/13) Doxycycline, 56% (39/70) Azithromycin and 0% (0/101) for Ciprofloxacin. PCR analysis suggested strains of V. cholerae O1 being descendants of the current seventh pandemic V. cholerae O1 CIRS 101 hybrid variant. The V. cholerae O1 currently causing cholera epidemics in north and central Mozambique confirmed a CTXΦ genotype and a molecular arrangement similar to the V. cholerae O1 CIRS 101. CONCLUSION: Although V. cholerae infections in Mozambique are generally not treated with antibiotics circulating strains of the bacteria showed high frequency of in vitro resistance to available antibiotics. Continuous monitoring of antibiotic resistance pattern of epidemic strains is therefore crucial since the appearance of antibiotic resistance can influence cholera control strategies.

Antibiotics resistance in El Tor Vibrio cholerae 01 isolated during cholera outbreaks in Mozambique from 2012 to 2015

Rationale: Mozambique has recorded cyclically epidemic outbreaks of cholera. Antibiotic therapy is recommended in specific situations for management and control of cholera outbreaks. However, an increase in resistance rates to antibiotics by Vibrio cholerae has been reported in several epidemic outbreaks worldwide. On the other hand, there are few recent records of continuous surveillance of antibiotics susceptibility pattern of V. cholerae in Mozambique. Goals: The purpose of this study was to evaluate antibiotics resistance pattern of Vibrio cholerae O1 Ogawa isolated during Cholera outbreaks in Mozambique to commonly used antibiotics. Methodology: We analyzed data from samples received in the context of surveillance and response to Cholera outbreaks in the National Reference Laboratory of Microbiology from the National Institute of Health of Mozambique, 159 samples suspected of cholera from cholera treatment centers of, Metangula (09), Memba (01), Tete City (08), Moatize (01), Morrumbala (01) districts, City of Quelimane (01), Lichinga (06) and Nampula (86) districts, from 2012 to 2015. Laboratory culture and standard biochemical tests were employed to isolate and identify Vibrio cholerae; serotypes were determined by antisera agglutination reaction in blade. Biotype and presence of important virulence factors analysis was done by PCR. Antibiotics susceptibility pattern was detected by disk diffusion method Kirby Bauer. Antibiotic susceptibility and results were interpreted by following as per recommendations of CLSI (Clinical and Laboratory Standards Institute) 2014. All samples were collected and tested in the context of Africhol Project, approved by the National Bioethics Committee for Health. Results: Among isolates from of Vibrio cholerae O1 El Tor Ogawa resistance to Sulphamethoxazole-trimethropim was 100% (53/53) to Trimethoprim-, being 100% (54/54) for Ampicillin, 99% (72/74) for Nalidixic Acid, 97% (64/66) to Chloramphenicol, 95% (42/44) for Nitrofurantoin and (19/20) Cotrimoxazole, 83% (80/97) Tetracycline, 56% (5/13) Doxycycline, 56% (39/70) Azithromycin and 0% (0/101) for Ciprofloxacin. PCR analysis suggested strains of V. cholerae O1 being descendants of the current seventh pandemic V. cholerae O1 CIRS 101 hybrid variant. The V. cholerae O1 currently causing cholera epidemics in north and central Mozambique confirmed a CTXΦ genotype and a molecular arrangement similar to the V. cholerae O1 CIRS 101. Conclusion: Although V. cholerae infections in Mozambique are generally not treated with antibiotics circulating strains of the bacteria showed high frequency of in vitro resistance to available antibiotics. Continuous monitoring of antibiotic resistance pattern of epidemic strains is therefore crucial since the appearance of antibiotic resistance can influence cholera control strategies.

Cholera Incidence and Mortality in Sub-Saharan African Sites during Multi-country Surveillance.

BACKGROUND: Cholera burden in Africa remains unknown, often because of weak national surveillance systems. We analyzed data from the African Cholera Surveillance Network (www.africhol.org). METHODS/ PRINCIPAL FINDINGS: During June 2011-December 2013, we conducted enhanced surveillance in seven zones and four outbreak sites in Togo, the Democratic Republic of Congo (DRC), Guinea, Uganda, Mozambique and Cote d'Ivoire. All health facilities treating cholera cases were included. Cholera incidences were calculated using culture-confirmed cholera cases and culture-confirmed cholera cases corrected for lack of culture testing usually due to overwhelmed health systems and imperfect test sensitivity. Of 13,377 reported suspected cases, 34% occurred in Conakry, Guinea, 47% in Goma, DRC, and 19% in the remaining sites. From 0-40% of suspected cases were aged under five years and from 0.3-86% had rice water stools. Within surveillance zones, 0-37% of suspected cases had confirmed cholera compared to 27-38% during outbreaks. Annual confirmed incidence per 10,000 population was <0.5 in surveillance zones, except Goma where it was 4.6. Goma and Conakry had corrected incidences of 20.2 and 5.8 respectively, while the other zones a median of 0.3. During outbreaks, corrected incidence varied from 2.6 to 13.0. Case fatality ratios ranged from 0-10% (median, 1%) by country. CONCLUSIONS/SIGNIFICANCE: Across different African epidemiological contexts, substantial variation occurred in cholera incidence, age distribution, clinical presentation, culture confirmation, and testing frequency. These results can help guide preventive activities, including vaccine use.

Epidemic waves of cholera in the last two decades in Mozambique.

INTRODUCTION: Africa is increasingly affected by cholera. In Mozambique, cholera appeared in the early 1970s when the seventh pandemic entered Africa from the Indian subcontinent. In the following decades, several epidemics were registered in the country, the 1997-1999 epidemic being the most extended. Since then, Mozambique has been considered an endemic area for cholera, characterized by yearly outbreaks occurring with a seasonal pattern. At least three pandemic variants are thought to have originated in the Indian subcontinent and spread worldwide at different times. To understand the epidemiology of cholera in Mozambique, whether the disease re-emerges periodically or is imported by different routes of transmission, we investigated clinical V. cholerae O1 isolated during 1997-1999 and 2012-2014 epidemics. METHODOLOGY: By detecting and characterizing seven genetic elements, the mobilome profile of each isolate was obtained. By comparing it to known seventh pandemic reference strains, it was possible to discern among different V. cholerae O1 variants active in the country. RESULTS: During 1997-1999, epidemic strains showed two different genetic profiles, both related to a pandemic clone that originated from India and was reported in other African countries in the 1990s. Isolates from 2012-2014 outbreaks showed a genetic background related to the pandemic strains currently active as the prevalent causative agent of cholera worldwide. CONCLUSIONS: Despite cholera being endemic in Mozambique, the epidemiology of the disease in the past 20 years has been strongly influenced by the cholera seventh pandemic waves that originated in the Indian subcontinent.