Intestinal protozoa in hospitalized under-five children with diarrhoea in Nampula - a cross-sectional analysis in a low-income setting in northern Mozambique.

BACKGROUND: In Mozambique, infection by intestinal parasites is reported all over the country. However, infection in children with diarrhoea is mostly focused in the southern region of Mozambique. This work aims to determine the frequency and potential risk factors for infection by Cryptosporidium spp., Giardia lamblia, and Entamoeba histolytica in children under-five years hospitalized with diarrhoea in Hospital Central de Nampula, northern Mozambique. METHODS: A cross-sectional hospital-based surveillance was conducted between March 2015 and January 2018 in children admitted with diarrhoea in Hospital Central de Nampula. Sociodemographic information was obtained through semi-structured interviews applied to the children's caregivers. A single stool sample was collected from each child to detect antigens from Cryptosporidium spp., G. lamblia, and E. histolytica using an immune-enzymatic technique. Crude and adjusted odds ratios (with 95% Confidence Intervals) were obtained by logistic regression models to identify factors associated with infection by Cryptosporidium spp. and G. lamblia. RESULTS: The median age and interquartile intervals of our sample population was 12 months (8-20). Intestinal protozoa were detected in 21.4% (59/276). Cryptosporidium spp. was the most common protozoa (13.9% - 38/274), followed by G. lamblia (9.1% - 25/274) and E. histolytica (0.4% - 1/275). Children with illiterate caregiver's (p-value = 0.042) and undernourished (p-value = 0.011) were more likely to be infected by Cryptosporidium spp. G. lamblia was more common in children living in households with more than four members (p-value = 0.039). E. histolytica was detected in an eleven month's child, co-infected with Cryptosporidium spp. and undernourished. CONCLUSION: Cryptosporidium spp. and Giardia lamblia were the most common pathogenic intestinal protozoa detected in children with diarrhoea hospitalized in the Hospital Central de Nampula. Our findings obtained highlight the importance of exploring the caregiver's education level, children's nutritional status for infections with Cryptosporidium spp., and living conditions, namely crowded households for infections with G. lamblia in children younger than five years.

Rotavirus A infection in children under five years old with a double health problem: undernutrition and diarrhoea - a cross-sectional study in four provinces of Mozambique.

BACKGROUND: Mozambique has a high burden of group A rotavirus (RVA) infection and chronic undernutrition. This study aimed to determine the frequency and potential risk factors for RVA infection in undernourished children under 5 years old with diarrhoea in Mozambique. METHODS: The analysis was conducted using data from March 2015 to December 2017, regarding children under 5 years old with at least one type of undernutrition. Anthropometric measures were used to calculate indices of weight-for-age, weight-for-height and height-for-age through the Z-Scores. RVA results were extracted from the National Diarrhoea Surveillance database. Descriptive statistics, chi-square test was used for qualitative variables and organized in contingency tables and 95% Confidence Intervals (CI) were considered for the calculation of RVA infection proportion and in the multiple logistic regression models to estimate the adjusted odds ratios (AOR). RESULTS: Of the 842 undernourished children included in the analysis, 27.2% (95% CI: 24.3-30.3%) were positive for RVA. The rate of RVA infection was 42.7% (95% CI: 38.0-47.5%) in the pre-vaccine period, with great reduction to 12.2% (95% CI: 9.4-15.6%) in the post-vaccine period. Most of the RVA undernourished children had severe wasting (33.3%) and severe stunting (32.0%). The risk of infection was significantly high in children from 0 to 11 months (p-value < 0.001) when compared to the age group of 24-59 months. A higher proportion of RVA infection was detected in households with five or more members (p-value = 0.029). Similar proportions of RVA were observed in children fed only by breast milk (34.9%) and breast milk with formula (35.6%). A higher proportion of undernourished HIV-positive children co-infected with RVA (7.4%) was observed. CONCLUSIONS: The frequency of RVA infection in undernourished children declined following the introduction of the vaccine in Mozambique. Beyond the temporal variation, Maputo province, age and crowded households were also associated to RVA infection. A high proportion of RVA infection was observed in children with severe wasting and a triple burden of disease: undernutrition, RVA and HIV, highlighting the need to conduct follow-up studies to understand the long-term impact of these conditions on children's development.

Enterovirus detection in stool samples from Mozambican children with acute gastroenteritis.

Enteroviruses (EV) are predominantly enteric viruses, present in all parts of the world causing disease in humans with a broad spectrum of clinical presentations. The purpose of this study was to identify non-polio enteroviruses (NPEV) in stool samples collected from children with acute gastroenteritis (AGE) symptoms of unknown etiology in four provinces (Maputo, Nampula, Sofala and Zambézia) of Mozambique. From June 2014 to March 2018, 327 stool samples were collected from children hospitalized with AGE in health care units. NPEVs were detected in 52 samples (52/327; 15.9%) and were more frequent in children under 5 years of age. The age group from 12 to 23 months was the most affected and showed more severity of disease. We also identified 26 different EV-types with the following detection pattern EV-B>EV-C>EV-A. The major EV-types were EV-A119 (9/52; 17.3%) and EV-C99 (8/52; 15.4%), accounting for 32.7% of the total. In addition to EV-A119, other uncommon EV-types were also identified, such as EV-B75, EV-B97 and EV-C113. The current study shows a high heterogeneity of EV types circulating in children with AGE in Mozambique as well as the identification of rarely described enteroviruses.

Rotavirus A infection in pre- and post-vaccine period: Risk factors, genotypes distribution by vaccination status and age of children in Nampula Province, Northern Mozambique (2015-2019).

Mozambique introduced the monovalent rotavirus vaccine (Rotarix®, GSK Biologicals, Rixensart, Belgium) in September 2015. Previous analysis, showed that Nampula province continues reporting a high frequency of Rotavirus A (RVA) infection and the emergence of G9P[6], G9P[4] and G3P[4] genotypes. This analysis aimed to determine the RVA frequency; risk factors; genotype distribution by vaccination status and age between pre- and post-vaccine periods in children under-five years old with diarrhea in Nampula. A cross-sectional, hospital-based surveillance study was conducted in the Hospital Central de Nampula in Mozambique. Socio-demographic and clinical data were collected to assess factors related to RVA infection in both periods. Stool specimens were screened to detect RVA by ELISA, and positive samples were genotyped. Between 2015 (pre-vaccine period) and 2016-2019 (post-vaccine period), 614 stool specimens were collected and tested for RVA in which 34.9% (67/192) were positive in pre-vaccine period and 21.8% (92/422) in post-vaccine (p = 0.001). In the post-vaccine period, age, year, and contact with different animal species (chicken, duck, or multiple animals) were associated with RVA infection. RVA infection was higher in children partially vaccinated (40.7%, 11/27) followed by the fully vaccinated (29.3%, 56/191) and the unvaccinated (15.3%, 21/137) (p = 0.002). G1P[8] and G9P[4] were common in vaccinated children less than 12 months. The present analysis showed that RVA infection reduced slightly in the post-vaccine period, with a high proportion of infection and genotype diversity in children, under 12 months of age, vaccinated. Further research on factors associated with RVA infection on vaccinated compared to unvaccinated children and vaccination optimization should be done.

Early impact of rotavirus vaccination in children less than five years of age in Mozambique.

BACKGROUND: Mozambique introduced rotavirus vaccine (Rotarix, GSK Biologicals) in the National Immunization Program in September 2015 with the objective of reducing the burden of total diarrheal disease and specifically severe rotavirus disease. This study aimed to evaluate the early impact of rotavirus vaccine in reducing all-cause diarrhea and rotavirus-specific hospitalizations. METHODS: We analysed stool specimens collected from children under five years old, between January 2014 and June 2017 within the National Surveillance for Acute Diarrhea. We compared annual changes in rotavirus positivity, median age of children hospitalized for rotavirus and the number of all-cause for diarrheal hospitalizations. Rotavirus detection was performed using enzyme immunoassay. RESULTS: During this period, 1296 samples were collected and analyzed. Rotavirus positivity before vaccine introduction was 40.2% (39/97) in 2014 and 38.3% (225/588) in 2015, then after vaccine introduction reduced to 12.2% and 13.5% in 2016 and 2017, respectively. The median age of children hospitalized for rotavirus was 9 and 11 months in 2014 and 2015 and 10 months in 2016 and 2017. Rotavirus hospitalizations exhibited a seasonal peak prior to vaccine introduction, between June and September in 2014 and 2015, coinciding with winter period in Mozambique. After vaccine introduction, the peak was delayed until August to December in 2016 and was substantially diminished. There was a reduction in all-cause acute diarrhea hospitalizations in children aged 0-11 months after vaccine introduction. CONCLUSION: We observed a reduction in rotavirus positivity and in the number of all-cause diarrhea hospitalizations after vaccine introduction. The data suggest rotavirus vaccine is having a positive impact on the control of rotavirus diarrheal disease in Mozambique.

Enterovirus detection in stool samples from Mozambican children with acute gastroenteritis

Enteroviruses (EV) are predominantly enteric viruses, present in all parts of the world causing disease in humans with a broad spectrum of clinical presentations. The purpose of this study was to identify non-polio enteroviruses (NPEV) in stool samples collected from children with acute gastroenteritis (AGE) symptoms of unknown etiology in four provinces (Maputo, Nampula, Sofala and Zambézia) of Mozambique. From June 2014 to March 2018, 327 stool samples were collected from children hospitalized with AGE in health care units. NPEVs were detected in 52 samples (52/327; 15.9%) and were more frequent in children under 5 years of age. The age group from 12 to 23 months was the most affected and showed more severity of disease. We also identified 26 different EV-types with the following detection pattern EV-B>EV-C>EV-A. The major EV-types were EV-A119 (9/52; 17.3%) and EV-C99 (8/52; 15.4%), accounting for 32.7% of the total. In addition to EV-A119, other uncommon EV-types were also identified, such as EV-B75, EV-B97 and EV-C113. The current study shows a high heterogeneity of EV types circulating in children with AGE in Mozambique as well as the identification of rarely described enteroviruses.

Microscopia e Ensaio de Imunoabsorção Enzimática: detecção de Cryptosporidium spp. em fezes de crianças moçambicanas / Microscopy and Enzyme-Linked Immunosorbent Assay for Cryptosporidium spp. detection in diarrheic feces of Mozambican children

A diarreia causada por Cryptosporidium spp. é comum em crianças e culmina com elevadas taxas de morbi-mortalidade, sobretudo nos países em desenvolvimento. A técnica convencional para o diagnóstico de Cryptosporidium spp. é a microscopia óptica, contudo, o Ensaio de Imunoabsorção Enzimática (ELISA) também tem sido usado. O objectivo deste estudo foi de comparar a performance do ELISA (ensaio comercial) em relação a microscopia óptica na detecção de Cryptosporidium spp. em fezes diarreicas de crianças admitidas no Hospital Geral de Mavalane (HGM) de Maio de 2014 a Fevereiro de 2015. No total, 130 amostras de fezes foram testadas, primeiramente por microscopia óptica e depois por ELISA. A estatística descritiva, sensibilidade, especi cidade e valores preditivos negativo e positivo foram usados para analisar os dados.A microscopia permitiu a identi cação de Cryptosporidium spp., Entamoeba hystolitica/díspar/moshovskii, Giardia intestinalis, Ascaris lumbricóides, Trichuris trichiura e Balantidium coli. Doze (9.2%) amostras foram positivas para Cryptosporidium spp., contra 22 (16.9%) identi cadas por ELISA. Esta diferença foi estatisticamente signi cativa (p = 0.002). A frequência de detecção deste patógeno por microscopia foi maior em crianças dos 12 aos 23 meses (4.6%), enquanto por ELISA foi maior em crianças dos zero aos 11 meses. A sensibilidade do ELISA na detecção de antígenos de Cryptosporidium spp. foi de 58.3%, enquanto a especi-cidade foi de 81.4%. Os resultados encontrados ressaltam a necessidade de uma ferramenta de diagnóstico complementar tal como o ELISA, para con rmar o diagnóstico em pacientes com sinais clínicos típicos de criptosporidiose mas com resultado negativo por microscopia.

Diarrhea caused by Cryptosporidium spp. is common in children and ends with high morbidity and mortality rates, mostly in developing countries. e conventional technique for the diagnosis of Cryptosporidium spp. is the optical microscopy, however, the Enzyme-linked immunosorbent assay (ELISA) is also being implemented. e aim of this study was to compare the performance of ELISA (commercial immunoassay) in relation to optical microscopy to detect Cryptosporidium spp. in diarrheic feces of children admitted to Mavalane General Hospital from May 2014 to February 2015. Overall 130 stool samples were tested rstly by optical microscopy and then by ELISA. Descriptive statistics, sensitivity, speci city, negative and positive predictive value were used to analyze the data. e microscopy detected 12 (9.2%) Cryptosporidiumspp., positive samples and ELISA identi ed 22 (16.9%). is di erence was statistically signi cant (p = 0.002). e frequency of detection by microscopy was higher in children from 12 to 23 months (4.6%), while by ELISA was higher in those from zero to 11 months. e microscopy enabled the identi cation of other parasites such as Entamoeba hystolitica/díspar/moshovskii, Giardia intestinalis, Ascaris lumbricóides, Trichuris trichiura and Balantidium coli. e sensitivity of the ELISA in detection of Cryptosporidium spp. antigens was 58.3%, while the speci city was 81.4%. Taking all together, our results highlight the need of a complementary diagnostic tool, such as this ELISA, to con rm the diagnosis in patients with typical clinicalsymptoms of cryptosporidiosis but negative results by microscopy.

Early impact of rotavirus vaccination in children less than five years of age in Mozambique

Mozambique introduced rotavirus vaccine (Rotarix, GSK Biologicals) in the National Immunization Program in September 2015 with the objective of reducing the burden of total diarrheal disease and specifically severe rotavirus disease. This study aimed to evaluate the early impact of rotavirus vaccine in reducing all-cause diarrhea and rotavirus-specific hospitalizations....

Detecção de vírus gastroentéricos e caracterização molecular dos rotavírus em crianças hospitalizadas com diarreia aguda e comorbidades na cidade de Maputo, Moçambique

As gastroenterites virais estão entre as mais importantes causas de morbidade e mortalidade em crianças com menos de cinco anos de idade, principalmente nos países em desenvolvimento. Este estudo teve como objetivos i) descrever a frequência de detecção de rotavírus do grupo A (RV-A), norovírus (NoV) e astrovírus humanos (HAstV); ii) caracterizar molecularmente os RV-A. Para tal, foram coletadas 123 amostras fecais de crianças no Hospital Geral de Mavalane, cidade de Maputo, Moçambique, entre novembro 2011 e fevereiro de 2012. Os testes laboratoriais foram realizados no Laboratório de Virologia Comparada e Ambiental (LVCA). O RV-A foi detectada por electroforese em gel de poliacrilamida (PAGE) e ensaio imunoenzimático (EIE), enquanto NoV e HAstV foram detectadas por RT-PCR. A caracterização molecular de G-tipo dos RV-A foi realizado pelo sequenciamento da VP7 e P-tipo através de semi-nested PCR multiplex Todas crianças foram hospitalizadas com diarréia, entretanto, a maioria também apresentava outros quadros infecciosos (comorbidades), como a malária (23,6%), infecções respiratórias agudas (21,1%), HIV / SIDA (13,8%) e desnutrição protéico-calórica (30,1%). Os agentes virais foram detectados em 27 (22%) das crianças estudadas. RV-A, NoV e HAstV foram detectadas em 4 (3,3%), 13 (10,6%), e 10 (8,1%) crianças, respectivamente. Das treze amostras positivas para NoV, oito foram classificadas como sendo do genogrupo II (GII) e uma do genogrupo I (GI). Três amostras positivas para RV-A foram genotipadas, duas foram caracterizadas como G12P[6] e outra como G12P[8]. A alta frequência de comorbidades pode explicar a detecção relativamente baixa de agentes virais, especialmente RV-A. A presença de RV-A G12 em Moçambique ilustra a emergência deste genótipo, como observado em outros países. Este é o primeiro estudo que aborda a caracterização molecular de RV-A e a descrição da ocorrência de NoV e HAstV em Moçambique. Os resultados encontrados neste estudo demonstram a importância de se estabelecer uma vigilância ativa da síndrome diarréica no país.