Epidemiology of rotavirus infection in children in Blantyre, Malawi, 1997-2007.

Acute gastroenteritis caused by rotavirus infection is an important cause of morbidity and mortality among infants and young children in Africa. From 1997 through 2007, we enrolled 3740 children <5 years of age with acute gastroenteritis who received hospital care at the Queen Elizabeth Central Hospital in Blantyre, Malawi. Group A rotavirus was detected in fecal specimens by enzyme immunoassay. Rotavirus strains were characterized for VP7 (G) and VP4 (P) types with use of reverse-transcription polymerase chain reaction. Overall, rotavirus was detected in one-third of children. The median age of children with rotavirus gastroenteritis was 7.8 months, compared with 10.9 months for those without rotavirus in stool specimens (P > .001). Rotavirus circulated throughout the year, with the detection proportion greatest during the dry season (from May through October). A total of 15 single rotavirus strain types were detected during the study period, with genotypes P[8]G1, P[6]G8, P[4]G8, P[6]G1, P[8]G3, and P[6]G9 comprising 83% of all strains characterized. Serotype G12 was detected for the first time in Blantyre during the final 2 years of study. Zoonotic transmission and viral reassortment contributed to the rich diversity of strains identified. Current rotavirus vaccines have the potential to greatly reduce the rotavirus disease burden in Malawi, but they will be required to protect against a broad range of rotavirus serotypes in a young population with year-round rotavirus exposure.

Short postexposure prophylaxis in newborn babies to reduce mother-to-child transmission of HIV-1: NVAZ randomised clinical trial.

BACKGROUND: In sub-Saharan Africa, most women present late for delivery with unknown HIV status, which limits the use of intrapartum nevirapine to prevent mother-to-child transmission of HIV. We aimed to determine whether post-exposure prophylaxis of nevirapine plus zidovudine given to babies only reduced transmission of HIV more than did a regimen of nevirapine alone. METHODS: We randomly assigned 1119 babies of Malawian women with HIV-1 who presented late (ie, within 2 h of expected delivery) to either nevirapine alone or nevirapine and zidovudine. Both drugs were given immediately after birth: one dose of nevirapine (2 mg/kg weight) was given as a single dose; babies in the nevirapine plus zidovudine group also received zidovudine twice daily for 1 week (4 mg/kg weight). Infant HIV infection was determined at birth and at 6-8 weeks. Primary outcome was HIV infection in babies at 6-8 weeks in those not infected at birth. Analysis was by intention to treat. FINDINGS: The overall rate of mother-to-child transmission at 6-8 weeks was 15.3% in 484 babies who received nevirapine and zidovudine and 20.9% in 468 babies who received nevirapine only (p=0.03). At 6-8 weeks, in babies who were HIV negative at birth, 34 (7.7%) babies who had nevirapine and zidovudine and 51 (12.1%) who received nevirapine only were infected (p=0.03)-a protective efficacy of 36%. This finding remained after controlling for maternal viral load and other factors at baseline. Adverse events were mild and of similar frequency in the two groups. INTERPRETATION: Postexposure prophylaxis can offer protection against HIV infection to babies of women who missed opportunities to be counselled and tested before or during pregnancy. The nevirapine and zidovudine regimen is safe and easy to implement.

Zinc homeostasis in Malawian children consuming a high-phytate, maize-based diet.

BACKGROUND: Zinc deficiency in children is an important public health concern in the developing world, and the consumption of predominantly cereal-based diets with a high phytate content may contribute to the risk. The gastrointestinal tract plays a central role in absorbing and conserving zinc, yet it has not been carefully studied in such children. OBJECTIVE: This study investigated zinc homeostasis in healthy, free-living Malawian children with habitually high-phytate diets to better understand the role of the gastrointestinal tract. DESIGN: We evaluated zinc homeostasis in 10 children aged 2-5 y who were consuming a maize-based diet (phytate:zinc molar ratio of 23:1). Zinc stable isotopes were administered orally and intravenously. The tracer and tracee were measured in urine and feces. RESULTS: Endogenous fecal zinc was high in comparison with results for this measure in previous studies. Typical correlations seen in subjects consuming a low-phytate diet between total absorbed zinc, the size of the exchangeable zinc pool, and endogenous fecal zinc were not observed. Fractional absorption of zinc was 0.24. CONCLUSIONS: Zinc homeostasis was perturbed, particularly by large, endogenous fecal zinc losses, in this vulnerable population. The effects of interventions to improve zinc status, including dietary phytate reduction, on zinc homeostasis merit further study.

Urea production and leucine oxidation in malnourished children with and without acute infection.

We compared the kinetics of urea production and leucine oxidation in severely malnourished Malawian children. We tested the hypotheses that the rate of urea production was directly proportional to the rate of leucine oxidation and that the relationship between the two is altered by acute infection. Thirty-six marasmic children, aged 12 to 60 months, were enrolled; 26 had acute infection and 10 did not. The rates of urea and CO(2) production were estimated using primed, constant, intravenous stable isotope-labeled tracer infusions followed by intermittent sampling of breath and blood. The rate of urea production was greater in infected children when compared to uninfected children (169 +/- 85 v 105 +/- 44 micromol urea x kg(-1) x h(-1), P <.02). For children with and without infection, the rates of leucine oxidation and urea production were directly correlated (r = 0.49 and r = 0.74, respectively; P <.01), but the slopes of the regression lines were different. In uninfected children the degree of wasting was correlated with the rates of urea production and leucine oxidation (r = 0.67 and r = 0.48, respectively; P <.05). These data suggest that the rates of leucine oxidation and urea production are both measures of nitrogen catabolism, that acute infection alters the relationship between the two, and that less nitrogen is lost as urea in children with more wasting.

HIV infection among paediatric in-patients in Blantyre, Malawi.

To investigate the impact of HIV infection on hospital admission and death we studied children admitted to paediatric medical and surgical wards in Blantyre, Malawi, in March 2000. Unselected children whose parents or guardians consented to HIV testing of the child were recruited and HIV infection was determined by serology, with confirmation in children aged 15 months or less by PCR. We assessed the prevalence of HIV infection by age, clinical diagnosis and outcome of admission. Of 1064 admissions, 991 were tested for HIV infection, and 187 (18.9%) were infected. HIV was most common in children aged less than six months, 53 of 166 (32%). Parents of HIV-infected children were better educated, and more likely to have died, than those of uninfected children. Clinical symptoms and signs were not adequately sensitive or specific to be used for diagnosis of HIV. HIV was common in children with malnutrition (prevalence 40%), lower respiratory tract infection (29%) and sepsis (28%), and less prevalent among children with malaria (11%) or surgical admissions (11%). Almost 30% of HIV-infected children died, compared with 8.9% of uninfected children, and HIV-infected children constituted over 40% of in-patient deaths.

Nevirapine and zidovudine at birth to reduce perinatal transmission of HIV in an African setting: a randomized controlled trial.

CONTEXT: Antenatal counseling and human immunodeficiency virus (HIV) testing are not universal in Africa; thus, women often present in labor with unknown HIV status without receiving the HIVNET 012 nevirapine (NVP) regimen (a single oral dose of NVP to the mother at the start of labor and to the infant within 72 hours of birth). OBJECTIVE: To determine risk of mother-to-child transmission of HIV when either standard use of NVP alone or in combination with zidovudine (ZDV) was administered to infants of women tested at delivery. DESIGN, SETTING, AND PARTICIPANTS: A randomized, open-label, phase 3 trial conducted between April 1, 2000, and March 15, 2003, at 6 clinics in Blantyre, Malawi, Africa. The trial included all infants born to 894 women who were HIV positive, received NVP intrapartum, and were previously antiretroviral treatment-naive. Infants were randomly assigned to NVP (n = 448) and NVP plus ZDV (n = 446). Infants were enrolled at birth, observed at 6 to 8 weeks, and followed up through 3 to 18 months. The HIV status of 90% of all infants was established at 6 to 8 weeks. INTERVENTION: Mothers received a 200-mg single oral dose of NVP intrapartum and infants received either 2-mg/kg oral dose of NVP or NVP (same dose) plus 4 mg/kg of ZDV twice per day for a week. MAIN OUTCOME MEASURES: HIV infection of infant at birth and 6 to 8 weeks, and adverse events. RESULTS: The mother-to-child transmission of HIV at birth was 8.1% (36/445) in infants administered NVP only and 10.1% (45/444) in those administered NVP plus ZDV (P =.30). A life table estimate of transmission at 6 to 8 weeks was 14.1% (95% confidence interval [CI], 10.7%-17.4%) in infants who received NVP and 16.3% (95% CI, 12.7%-19.8%) in those who received NVP plus ZDV (P =.36). For infants not infected at birth and retested at 6 to 8 weeks, transmission was 6.5% (23/353) in those who received NVP only and 6.9% (25/363) in those who received NVP plus ZDV (P =.88). Almost all infants (99%-100%) were breastfed at 1 week and 6 to 8 weeks. Grades 3 and 4 adverse events were comparable; 4.9% (22/448) and 5.4% (24/446) in infants receiving NVP only and NVP plus ZDV, respectively (P =.76). CONCLUSIONS: The frequency of mother-to-child HIV transmission at 6 to 8 weeks in our 2 study groups was comparable with that observed for other perinatal HIV intervention studies among breastfeeding women in Africa. The safety of the regimen containing neonatal ZDV was similar to that of a standard NVP regimen.

Late postnatal transmission of HIV-1 and associated factors.

BACKGROUND: The present study was undertaken to determine the risk and timing of late postnatal transmission (LPT) of human immunodeficiency virus type 1 (HIV-1). METHODS: Breast-fed infants previously enrolled in 2 trials of antiretroviral prophylaxis were monitored in Malawi. Kaplan-Meier and proportional hazard models assessed cumulative incidence and association of factors with LPT. RESULTS: Overall, 98 infants were HIV infected, and 1158 were uninfected. The cumulative risk of LPT at age 24 months was 9.68% (95% confidence interval, 7.80%-11.56%). The interval hazards at 1.5-6, 6-12, 12-18, and 18-24 months were 1.22%, 4.05%, 3.48%, and 1.27%, respectively. CONCLUSIONS: The risk of LPT beyond 6 months is substantial. Weaning at 6 months could prevent >85% of LPT.

The impact of breastfeeding on the health of HIV-positive mothers and their children in sub-Saharan Africa.

OBJECTIVE: We assessed the impact of breastfeeding by women infected with human immunodeficiency virus (HIV)-1 on their morbidity and risk of mortality and on the mortality of their children. METHODS: We analysed longitudinal data from two previous randomized clinical trials of mother-to-child transmission of HIV conducted between April 2000 and March 2003 in the Republic of Malawi, Africa. Mothers infected with HIV, and their newborns, were enrolled at the time of their child's birth; they then returned for follow-up visits when the child was aged 1 week, 6-8 weeks and then 3, 6, 9, 15, 18, 21 and 24 months. Patterns of breastfeeding (classified as exclusive, mixed or no breastfeeding), maternal morbidity and mortality, and mortality among their children were assessed at each visit. Descriptive and multivariate analyses were performed to determine the association between breastfeeding and maternal and infant outcomes. FINDINGS: A total of 2000 women infected with HIV were enrolled in the original studies. During the 2 years after birth, 44 (2.2%) mothers and 310 (15.5%) children died. (Multiple births were excluded.) The median duration of breastfeeding was 18 months (interquartile range (IQR)=9.0-22.5), exclusive breastfeeding 2 months (IQR=2-3) and mixed feeding 12 months (IQR=6-18). Breastfeeding patterns were not significantly associated with maternal mortality or morbidity after adjusting for maternal viral load and other covariates. Breastfeeding was associated with reduced mortality among infants and children: the adjusted hazard ratio for overall breastfeeding was 0.44 (95% confidence interval (CI)=0.28-0.70), for mixed feeding 0.45 (95% CI=0.28-0.71) and for exclusive breastfeeding 0.40 (95% CI=0.22-0.72). These protective effects were seen both in infants who were infected with HIV and those who were not. CONCLUSION: Breastfeeding by women infected with HIV was not associated with mortality or morbidity; it was associated with highly significant reductions in mortality among their children.

Detection and characterization of human caliciviruses in hospitalized children with acute gastroenteritis in Blantyre, Malawi.

The human caliciviruses (HuCVs), including Norovirus and Sapovirus, are recognized causes of acute gastroenteritis in children and adults. A 1-year study was undertaken in Blantyre, Malawi, to examine the prevalence, and genetic diversity, of human caliciviruses (HuCVs) amongst children under 5 years of age hospitalized with acute gastroenteritis. Using the reverse transcription-polymerase chain reaction (RT-PCR), combined with nucleotide sequencing of the RT-PCR products, HuCVs were detected in 34/398 (8.5%) of children. Twelve (35.3%) of the children were co-infected with additional enteric viruses (predominantly rotavirus). The HuCVs comprised 26 Noroviruses (6.5%) and 8 Sapoviruses (2.0%). Each of the Noroviruses belonged to genogroup II, and could be further classified into six genotypes, including GII/3 (18 strains), GII/4 (2 strains), GII/11 (1 strain), GII/13 (1 strain), GII/16 (2 strains), and a putative new genotype GII/20 (2 strains). Each of the Sapoviruses belonged to genogroup GIII. HuCVs are the second most commonly identified viral enteropathogens (after rotavirus) among hospitalized children with gastroenteritis in Malawi.

Gender differences in perinatal HIV acquisition among African infants.

OBJECTIVE: We investigated gender-specific risks of mother-to-child transmission (MTCT) at birth and at 6 to 8 weeks among infants born to HIV-infected African women. DESIGN: Follow-up study of infants enrolled in 2 randomized, phase III, clinical trials to prevent MTCT, conducted in Blantyre, Malawi, in southeast Africa. METHODS: Infants were enrolled at birth and monitored postnatally, and their HIV status was assessed at birth and at 6 to 8 weeks (assessment beyond 6-8 weeks is ongoing). Statistical analyses were stratified according to gender, and comparisons were made with descriptive, univariate, and multivariate statistical tests. MTCT was estimated at birth and at 6 to 8 weeks among infants who were not infected at birth. RESULTS: Overall, 966 boys and 998 girls were enrolled. The rate of HIV transmission at birth was 9.5% (187 of 1964 infants). However, at birth significantly more girls (12.6%) than boys (6.3%) were infected with HIV. This association remained significant after controlling for maternal viral load and other factors. Among infants who were uninfected at birth, 8.7% (135 of 1554 infants) acquired HIV by 6 to 8 weeks; of these infants, more girls acquired HIV (10.0%), compared with boys (7.4%). CONCLUSIONS: Female infants may be more susceptible to HIV infection before birth and continuing after birth. Alternatively, in utero mortality rates of HIV-infected male infants may be disproportionately higher and thus more HIV-infected female infants are born. In areas of sub-Saharan Africa, where HIV infection rates are high among women of reproductive age, the magnitude of the gender transmission differences observed in this study could have clinical, preventive, and demographic implications.

Abnormal vitamin A cytology and mortality in infants aged 9 months and less with measles.

We investigated the relationship between abnormal vitamin A cytology and mortality in infants less than 9 months of age with measles. In a 12-month period, 116 children of this age with measles consecutively admitted to Queen Elizabeth Central Hospital (QECH) in Blantyre, Malawi were enrolled in the study. All guardians of patients were interviewed and clinical information and consent were obtained in a standardised fashion. Conjunctival impression cytology (CIC) was attempted on all and collected from 93 (80%) children. The overall mortality was 16%. The proportional hazards model revealed that the presence of pneumonia on admission (hazard ratio 9.58, p = 0.0002) and an abnormal vitamin A CIC on admission (hazard ratio 6.40, p = 0.003) were independently associated with mortality. Our findings suggest a relationship between abnormal vitamin A CIC and fatality in infants under 9 months of age.

Post-neonatal infant mortality in Malawi: the importance of maternal health.

In a cohort study of mothers and their infants, information was collected from women attending the antenatal services of two hospitals in a rural area of Malawi and 561 of their babies were enrolled in a follow-up study. There were 128 with a low birthweight (LBW, <2500 g), 138 with fetal anaemia (FA, cord haemoglobin <12.5 g/dl), 42 with both and 228 with a normal birthweight and no FA. Infants were seen monthly for 1 year. Risk factors for post-neonatal infant mortality (PNIM) were calculated using Cox regression analysis adjusting for LBW and FA. PNIM was 9.3%. Respiratory infections and diarrhoeal disease were the principal attributable causes of death. PNIM increased with LBW (RR 3.08, 95% CI 1.51-6.23) but not significantly so with FA (RR 1.60, 95% CI 0.78-3.27). An additional effect on PNIM was observed with maternal HIV (RR 3.44, 95% CI 1.63-7.26) and malaria at the first antenatal visit (RR 2.26, 95% CI 1.09-4.73). Illiteracy was not associated with mortality. Placental malaria in HIV-seronegative mothers was significantly associated with increased PNIM. Improving birthweight through effective antimalarial control in pregnancy will lead to a reduction in PNIM. Reduction of HIV prevalence and prevention of mother-to-child transmission of HIV must be a main target for government health policy.

Whole-body leucine kinetics and the acute phase response during acute infection in marasmic Malawian children.

This study compared leucine kinetics and acute-phase protein and cytokine concentrations in three groups of Malawian children who were fed an isoenergetic, isonitrogenous diet: children with marasmus with (n = 25) and without (n = 17) infection and well-nourished children with infection (n =13). The hypotheses tested were that whole-body leucine kinetics will be less in marasmic acutely infected children than in well-nourished acutely infected children but greater than in marasmic uninfected children. Children were studied after 24 h of therapy using standard (13)C-leucine stable isotope tracer techniques. Well-nourished children with acute infection had greater leucine kinetic rates than did marasmic children with acute infection; nonoxidative leucine disposal was 153 +/- 31 versus 118 +/- 43 micromol leucine. kg(-1). h(-1), leucine derived from whole-body proteolysis was 196 +/- 34 versus 121 +/- 47, and leucine oxidation was 85 +/- 31 versus 45 +/- 13 (p < 0.01 for all comparisons). Leucine kinetic rates were similar in marasmic children with and without acute infection. Well-nourished children with acute infection increased their serum concentration of five of six acute-phase proteins during the first 24 h, whereas marasmic children with infection did not have any increases. The serum concentrations of IL-6 were elevated in well-nourished and marasmic children with infection. These data suggest that the cytokine stimulus for the acute-phase protein kinetic response to acute infection is present in marasmic children but that the acute-phase protein metabolic response is blunted by malnutrition.

Community-based dietary phytate reduction and its effect on iron status in Malawian children.

This study describes a community-based method used in rural Malawi to remove dietary phytate, an inhibitor of iron absorption, and notes an improvement in the iron status of ten children who participated in the trial. Phytate was removed by soaking maize flour in excess water with phytase and decanting the water before cooking the flour. Iron status, as measured by soluble transferrin receptor and zinc protoporphyrin, was improved but not normal.

The risk of human immunodeficiency virus-1 infection in twin pairs born to infected mothers in Africa.

We examined birth order and delivery route as risk factors for mother-to-child transmission of human immunodeficiency virus (HIV)-1 in 315 twin pairs born in Malawi during 1994-1998. No antiretroviral drugs were administered to these subjects. Infections were detected by polymerase chain reaction and were stratified as having occurred either in utero, perinatally, or postnatally. Risk of in utero infection for 630 infants (39 infections) did not differ by birth order (first born, 6.3%; second born, 6.0%). Similarly, in 260 vaginally delivered infants evaluated for perinatal infection (45 infections), risk did not differ by birth order (first born, 15.9%; second born, 18.7%); risk of perinatal infection was significantly lower in cesarean-delivered infants (odds ratio, 0.19 [95% confidence interval, 0.02-0.78]). There was no effect on postnatal transmission rates. Thus, in contrast to the authors of earlier studies, we did not find birth order to be an important risk factor for infection in twins. These findings indicate that birth-canal exposure is not a major contributor to a newborn's risk of HIV-1 infection.

Molecular epidemiology of cryptosporidiosis in children in Malawi.

Few studies have examined the molecular epidemiology of cryptosporidiosis in developing countries. In this study, DNA of 69 microscopy-positive human fecal samples collected from Malawi were examined by multilocus genetic analyses. From 43, 27 and 28 of the samples, the SSU rRNA, 70 kDa heat shock protein (HSP70) and 60 kDa glycoprotein (GP60) genes, respectively, were successfully PCR-amplified. Restriction analysis of the SSU PCR products showed that 41 of the 43 PCR-positive samples had C. hominis and 2 had C. parvum. Sequence analysis of the HSP70 and GP60 gene confirmed the species identification by SSU rRNA PCR-RFLP analysis, but also revealed high intraspecific variations. Altogether, six HSP70 subtypes and six GP60 subtypes (belonging to four subtype alleles) of C. hominis were found. Linkage disequilibrum analysis of the two genetic loci showed possible intraspecific recombination. Thus, cryptosporidiosis in the study area was largely caused by anthroponotic transmission. The high intraspecific variation and existence of genetic recombination were probably results of high transmission of cryptosporidiosis in this area.

Promiscuous expression of Epstein-Barr virus genes in Burkitt's lymphoma from the central African country Malawi.

Primary BL in Malawian children has a very high frequency association, approaching 100%, with the human herpesvirus EBV. A detailed study carried out on viral gene expression in these tumours, using both fresh material and methanol-fixed FNAs, showed, contrary to prediction, that most belong to a variant "class II" latency category, with lytic cycle-related genes also expressed. That is, in addition to EBNA1 expression, membrane proteins (LMP1/2A), immediate early (BZLF1) and early (IR2 and IR4) genes, a putative viral oncogene (BARF1), CST (BART) antisense transcripts and the viral bcl-2 homologue are expressed in a high proportion of the BLs. Most, but not all, express the small viral (EBER) RNAs. Two other significant observations were made: (i) in addition to expression of cellular cytokine (IL-10) transcripts in all tumours investigated, the normally silent viral IL-10 homologue was expressed in some tumours; (ii) whereas EBNA1 expression from its restricted Qp promoter was generally observed, the nonrestricted Cp/Wp promoter was also active in some tumours. Viral gene expression in the Malawian [endemic (e)] BLs appears to be more promiscuous than predicted from other studies, but expression accords with the cytopathologic picture of eBLs as a rapidly proliferating cell population accompanied by considerable necrosis, and a clinically diverse disease. A small-scale study of relapse Malawian BLs revealed a different picture of viral association, more akin to systemic BL than eBL, where EBV appears to be absent or present only at very low levels. The significance of these findings is considered.

Detection and characterisation of human astroviruses in children with acute gastroenteritis in Blantyre, Malawi.

In a 2-year hospital-based study of paediatric gastroenteritis in Blantyre, Malawi, astroviruses were detected by enzyme immunoassay in 15 (1.9%) of 786 inpatients and in 9 (2.3%) of 400 outpatients. Greater disease severity was noted in children coinfected with human immunodeficiency virus (HIV). Six human astrovirus (HAstV) genotypes were identified, including HAstV-1 (25%), HAstV-2 (21%), HAstV-3 (25%), HAstV-4 (13%), HAstV-5 (4%), and HAstV-8 (13%). Although astroviruses are not major causes of gastroenteritis among children admitted to hospital in Blantyre, concomitant HIV infection appears to be a risk factor for increased severity of disease.