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

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.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.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.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.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

CTX ϕ cluster analysis of <i>Vibrio cholerae</i> O1 El Tor Ogawa isolated during cholera outbreaks in Mozambique from 2012 to 2015.

<p>Showing a classic signature of <i>Vibrio cholerae</i> O1 El Tor variants B33 and CIRS 101. AMP- Ampicillin; TE-Tetracycline; NA- Nalidixic Acid; C-Chloramphenicol; CIP-Ciprofloxacin; SXT- Sulphamethoxazol-trimethropim; F- Nitrofurantoin; AZM- Azithromycin; rstR, ctxB and tcpA- <i>Vibrio cholerae</i> virulence genes; TLC-RS1, CORE-RTX and TCL-RS2—primers for the presence of CTX ϕ on chromosome 1; Chr II—Chromosome 2.</p

AST per district and per year, for <i>V</i>. <i>cholerae</i> O1 El Tor Ogawa isolated during cholera outbreaks in Mozambique from 2012 to 2015.

<p>In 2012 (Cuamba district in Niassa province and Montepuez <i>district</i> in Cabo Delgado province), 2013 (Cuamba district in Niassa province, Pemba city and Montepuez <i>districts</i> in Cabo Delgado province, Nampula city <i>district</i> in Nampula province and Alto-Molócue <i>district</i> in Zambezia province), 2014 (Nampula city <i>district</i> in Nampula province), 2015 (Lichinga city, Lago and Cuamba <i>districts</i> in Niassa province, Nampula city <i>district</i> in Nampula province and Morrumbala and Quelimane city <i>districts</i> in Zambézia province, Tete city and Moatize <i>districts</i> in Tete province, Beira city <i>district</i> in Sofala province and Matola city <i>district</i> in Maputo province). AMP- Ampicillin; TE-Tetracycline; NA- Nalidixic Acid; C-Chloramphenicol; CIP-Ciprofloxacin; SXT- Sulphamethoxazol-trimethropim; F- Nitrofurantoin; AZM- Azithromycin; rstR, ctxB and tcpA- <i>Vibrio cholerae</i> virulence genes; TLC-RS1, CORE-RTX and TCL-RS2—primers for the presence of CTX ϕ on chromosome 1; Chr II—Chromosome 2</p

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

<div><p>Background</p><p>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.</p><p>Methodology/Principal findings</p><p>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 <i>Vibrio cholerae</i>, as well as clinical, behavioral, and socio-demographic variables. We analyzed factors associated with confirmed, hospitalized, and fatal cholera using multivariate logistic regression models.</p><p>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 <i>Vibrio cholerae</i> 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.</p><p>Conclusions/Significance</p><p>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.</p></div

Estimated cholera incidence rates by site, 2011–2015, Mozambique.

<p>Estimated cholera incidence rates by site, 2011–2015, Mozambique.</p

Epidemic curves, Africhol surveillance zones, Mozambique, 2011–2015: Beira City, Mocuba District, and Pemba City.

<p>Epidemic curves, Africhol surveillance zones, Mozambique, 2011–2015: Beira City, Mocuba District, and Pemba City.</p

Tested and confirmed cases of cholera through Africhol, Mozambique, 2011–2015.

<p>Tested and confirmed cases of cholera through Africhol, Mozambique, 2011–2015.</p

Africhol surveillance zones and outbreak sites in Mozambique.

<p>Note: Numbers in brackets are (cases, deaths).</p

Epidemic curves, Africhol outbreak sites, Mozambique, 2011–2015: Cuamba District, Montepuez District and Nampula City.

<p>Epidemic curves, Africhol outbreak sites, Mozambique, 2011–2015: Cuamba District, Montepuez District and Nampula City.</p

Socio-demographic characteristics of cholera suspected cases, 2011–2015, Mozambique.

<p>Socio-demographic characteristics of cholera suspected cases, 2011–2015, Mozambique.</p