Characterizing Undernourished Children Under-Five Years Old with Diarrhoea in Mozambique: A Hospital Based Cross-Sectional Study, 2015­2019

Diarrhoea is associated with undernutrition and this association is related to increased morbidity and mortality in children under-five. In this analysis we aimed to assess the frequency and associated factors of undernutrition in children under-five with diarrhoea. A hospital-based cross-sectional study was conducted from January 2015 to December 2019 through a surveillance system in five sentinel hospitals in Mozambique. Sociodemographic and clinical information was collected, including anthropometry. A total of 963 children were analysed. The overall undernutrition frequency was 54.1% (95% CI: 50.9­57.2), with 32.5% (95% CI: 29.6­35.5) stunting, 26.6% (95% CI: 23.9­29.6) wasting and 24.7% (95% CI: 22.1­27.5) underweight. Children from Nampula province had 4.7 (p = 0.016) higher odds for stunting compared with children from Maputo. Children whose caregiver was illiterate had higher odds of being underweight 5.24 (p < 0.001), and the wet season was associated with higher odds 1.70 (p = 0.012) of being wasted. Children born under 2500 g of weight had 2.8 (p = 0.001), 2.9 (p < 0.001) and 2.6 (p = 0.010) higher odds for being underweighted, wasted and stunted, respectively. The HIV positive status of the children was associated with higher odds of being underweight 2.6 (p = 0.006), and stunted 3.4 (p = 0.004). The province, caregiver education level, wet season, child's birthweight and HIV status were factors associated with undernutrition in children with diarrhoea. These findings emphasise the need for additional caregiver's education on the child's nutrition and associated infectious diseases. More studies are needed to better understand the social context in which a child with diarrhoea and undernutrition is inserted.

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...

Molecular epidemiology of rotavirus a strains pre- and post-vaccine (rotarix®) introduction in mozambique, 2012­2019: emergence of genotypes g3p[4] and g3p[8]

Group A rotavirus (RVA) remains the most important etiological agent associated with severe acute diarrhea in children. Rotarix® monovalent vaccine was introduced into Mozambique's Expanded Program on Immunization in September 2015. In the present study, we report the diversity and prevalence of rotavirus genotypes, pre- (2012­2015) and post-vaccine (2016­2019) introduction in Mozambique, among diarrheic children less than five years of age. Genotyping data were analyzed for five sentinel sites for the periods indicated. The primary sentinel site, Mavalane General Hospital (HGM), was analyzed for the period 2012­2019, and for all five sites (country-wide analyses), 2015­2019. During the pre-vaccine period, G9P[8] was the most predominant genotype for both HGM (28.5%) and the country-wide analysis (46.0%). However, in the post-vaccine period, G9P[8] was significantly reduced. Instead, G3P[8] was the most common genotype at HGM, while G1P[8] predominated country-wide. Genotypes G9P[4] and G9P[6] were detected for the first time, and the emergence of G3P[8] and G3P[4] genotypes were observed during the post-vaccine period. The distribution and prevalence of rotavirus genotypes were distinct in pre- and post-vaccination periods, while uncommon genotypes were also detected in the post-vaccine period. These observations support the need for continued country-wide surveillance to monitor changes in strain diversity, due to possible vaccine pressure, and consequently, the effect on vaccine effectiveness.

Prevalência de Parasitoses Intestinais em Crianças nas Escolas Primárias da Província de Sofala, Moçambique / Prevalence of Intestinal Parasitosis in Primary School Children in Sofala Province Province, Mozambique

Introdução: O complexo de infecções intestinais parasitárias é causado por organismos que podem ser biologicamente classificados em protozoários ou helmintos. Os parasitas constituem o maior grupo de organismos responsável por infecções em humanos. A província de Sofala localiza-se na região centro de Moçambique e está dividida em 13 distritos (incluindo a Cidade da Beira). O presente estudo foi realizado com o objectivo de determinar a prevalência de parasitoses intestinais nas escolas primárias dos distritos da província de Sofala. Métodos: O estudo decorreu de Agosto de 2005 a Junho de 2007 em escolas primárias dos distritos da província de Sofala. Tratou-se de um estudo transversal, de amostragem multietápica que teve como alvo crianças em idade escolar. Foram colhidas amostras de fezes e estas foram processadas por técnicas de Ritchie e Kato-Katz. O estudo foi aprovado pelo Comité Nacional de Bioética para Saúde de Moçambique e todos os princípios éticos foram seguidos. Os dados foram analisados utilizando o pacote estatístico SPSS na sua versão 15.0 e o software Microsoft Office Excel 2007. Resultados: Foram envolvidos um total de 5.748 alunos. A prevalência global de parasitoses ao nível da província foi de 34,4%. O distrito de Muanza foi a mais prevalente com 59,4% e o distrito de Dondo a menos prevalente com 14,1%. As espécies Entamoeba coli (23,6%), T. trichiura (16,0%), A. duodenale (14,0%) e A. lumbricoides (12,7%) foram as mais prevalentes. Observaram-se diferenças estatisticamente significativas em relação a idade e o nível de escolaridade e à prevalência de parasitas intestinais (p <0,001). Discussão e Conclusões: A prevalência de parasitoses intestinais em crianças nas escolas primárias da província de Sofala foi elevada. A presença Entamoeba coli é um indicativo epidemiológico de ocorrência de contaminação fecal. Há necessidade de se realizar estudos mais detalhados considerando as características e disposições ambientais onde elas vivem.

Diferenças de género e sexo na susceptibilidade às doenças infecciosas / Gender and sex difference susceptibility to infectious diseases

As doenças infecciosas ocorrem de forma desproporcional entre homens e mulheres. Muitas vezes, a severidade e o resultado da doença estão geralmente relacionados ao sexo. Os factores inerentes a estas diferenças ainda estão a ser estudados. Contudo, alguns estudos apontam o sexo como um dos factores principais das difer- enças da resposta imunológica às infecções. As mulheres desenvolvem naturalmente uma imunidade inata, celular e humoral com algumas utuações devido aos níveis hormonais que naturalmente variam durante os ciclos menstruais, gravidez, menopausa ou pelo uso de anticonceptivos hormonais ou o uso de tratamentos hormonais. Entretanto, as hormonas masculinas também têm sido implicadas na imunoregulação. A testesterona por exemplo causa o declínio de certas respostas imunológicas e os indivíduos jovens do sexo masculino são muitas vezes susceptíveis a infecções severas comparados às mulheres ou a homens adultos. Por outro lado, o género joga um papel importante na susceptibilidade às infecções transmitidas por vectores. Outros estudos apontam que o padrão hormonal no seio da família e a química da pele in uenciam na susceptibilidade à picada por mosquitos e outros vectores nos homens e mulheres. As diferenças comportamentais, ocupacionais e culturais também estão relacionadas com a exposição a vectores, assim como o uso de medidas de protecção individual e a procura pelos cuidados de saúde. Tomando em conta estes aspectos, é importante que futuros estudos imunológicos, epidemiológi- cos, comportamentais, de conhecimentos atitudes e práticas tenham em conta o sexo como uma variável, para uma melhor interpretação dos resultados e melhores práticas de saúde, princi- palmente no que tange ao tratamento das doenças infecciosas.

Infectious diseases occur di erently in men and women. Usually the severity and the result of the disease are related to the gender. Although some studies indicate the gender as one of main factor of immunological di erences, the causes of these di erences are still to be revealed. However, Women naturally develop innate immune response, cellular and humoral response with some uctuations due to the level of hormones that varies during the menstrual cycle, pregnancy and menopause, or due to the use of anti-conception pill or the use of hormonal treatments. e hormones from males also are implicated in immunoregulation. Testosterone decreases certain immune responses and young males are more susceptible to severe infections compared to adult males and females. In other hand, the gender was shown to play an important role on the susceptibility to vector transmitted diseases where the hormonal pattern within the family and the chemistry of the skin in uence the susceptibility to mosquito's bite and other vectors in men and women. e behavior, occupational and cultural behaviors in uence the exposure to vectors, but also the use of individual protection measures and the search for health services. Taking together, it's very important that future studies address gender and sex di erences in their designs, in order to conduct better conclusions and heath practices related to infectious diseases treatment or prevention.

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....

Norovirus Genetic Diversity in Children under Five Years Old with Acute Diarrhea in Mozambique (2014­2015)

Norovirus (NoV) is the second most important cause of viral diarrheal disease in children worldwide after rotavirus and is estimated to be responsible for 17% of acute diarrhea in low-income countries. This study aimed to identify and report NoV genotypes in Mozambican children under the age of five years with acute diarrhea. Between May 2014 and December 2015, stool specimens were collected within the Mozambique Diarrhea National Surveillance (ViNaDia) and tested for NoV genogroups I (GI) and II (GII) using conventional reverse transcriptase-polymerase chain reaction (RT-PCR). Partial capsid and RNA-dependent RNA polymerase (RdRp) nucleotide sequences were aligned using the Muscle tool, and phylogenetic analyses were performed using MEGA X. A total of 204 stool specimens were tested for NoV. The detection rate of NoV was 14.2% (29/204). The presence of NoV was confirmed, by real-time RT-PCR (RT-qPCR), in 24/29 (82.8%) specimens, and NoV GII predominated (70.8%; 17/24). NoV GII.4 Sydney 2012[P31] was the predominant genotype/P-type combination detected (30.4%; 7/23). This is the first study which highlights the high genetic diversity of NoV in Mozambican children and the need to establish a continuous NoV surveillance system.