Rotavirus genotypes in children under five years hospitalized with diarrhea in low and middle-income countries: Results from the WHO-coordinated Global Rotavirus Surveillance Network.

Rotavirus is the most common pathogen causing pediatric diarrhea and an important cause of morbidity and mortality in low- and middle-income countries. Previous evidence suggests that the introduction of rotavirus vaccines in national immunization schedules resulted in dramatic declines in disease burden but may also be changing the rotavirus genetic landscape and driving the emergence of new genotypes. We report genotype data of more than 16,000 rotavirus isolates from 40 countries participating in the Global Rotavirus Surveillance Network. Data from a convenience sample of children under five years of age hospitalized with acute watery diarrhea who tested positive for rotavirus were included. Country results were weighted by their estimated rotavirus disease burden to estimate regional genotype distributions. Globally, the most frequent genotypes identified after weighting were G1P[8] (31%), G1P[6] (8%) and G3P[8] (8%). Genotypes varied across WHO Regions and between countries that had and had not introduced rotavirus vaccine. G1P[8] was less frequent among African (36 vs 20%) and European (33 vs 8%) countries that had introduced rotavirus vaccines as compared to countries that had not introduced. Our results describe differences in the distribution of the most common rotavirus genotypes in children with diarrhea in low- and middle-income countries. G1P[8] was less frequent in countries that had introduced the rotavirus vaccine while different strains are emerging or re-emerging in different regions.

The Global Landscape of Pediatric Bacterial Meningitis Data Reported to the World Health Organization-Coordinated Invasive Bacterial Vaccine-Preventable Disease Surveillance Network, 2014-2019.

BACKGROUND: The World Health Organization (WHO) coordinates the Global Invasive Bacterial Vaccine-Preventable Diseases (IB-VPD) Surveillance Network to support vaccine introduction decisions and use. The network was established to strengthen surveillance and laboratory confirmation of meningitis caused by Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria meningitidis. METHODS: Sentinel hospitals report cases of children <5 years of age hospitalized for suspected meningitis. Laboratories report confirmatory testing results and strain characterization tested by polymerase chain reaction. In 2019, the network included 123 laboratories that follow validated, standardized testing and reporting strategies. RESULTS: From 2014 through 2019, >137 000 suspected meningitis cases were reported by 58 participating countries, with 44.6% (n = 61 386) reported from countries in the WHO African Region. More than half (56.6%, n = 77 873) were among children <1 year of age, and 4.0% (n = 4010) died among those with reported disease outcome. Among suspected meningitis cases, 8.6% (n = 11 798) were classified as probable bacterial meningitis. One of 3 bacterial pathogens was identified in 30.3% (n = 3576) of these cases, namely S. pneumoniae (n = 2177 [60.9%]), H. influenzae (n = 633 [17.7%]), and N. meningitidis (n = 766 [21.4%]). Among confirmed bacterial meningitis cases with outcome reported, 11.0% died; case fatality ratio varied by pathogen (S. pneumoniae, 12.2%; H. influenzae, 6.1%; N. meningitidis, 11.0%). Among the 277 children who died with confirmed bacterial meningitis, 189 (68.2%) had confirmed S. pneumoniae. The proportion of pneumococcal cases with pneumococcal conjugate vaccine (PCV) serotypes decreased as the number of countries implementing PCV increased, from 77.8% (n = 273) to 47.5% (n = 248). Of 397 H. influenzae specimens serotyped, 49.1% (n = 195) were type b. Predominant N. meningitidis serogroups varied by region. CONCLUSIONS: This multitier, global surveillance network has supported countries in detecting and serotyping the 3 principal invasive bacterial pathogens that cause pediatric meningitis. Streptococcus pneumoniae was the most common bacterial pathogen detected globally despite the growing number of countries that have nationally introduced PCV. The large proportions of deaths due to S. pneumoniae reflect the high proportion of meningitis cases caused by this pathogen. This global network demonstrated a strong correlation between PCV introduction status and reduction in the proportion of pneumococcal meningitis infections caused by vaccine serotypes. Maintaining case-based, active surveillance with laboratory confirmation for prioritized vaccine-preventable diseases remains a critical component of the global agenda in public health.The World Health Organization (WHO)-coordinated Invasive Bacterial Vaccine-Preventable Disease (IB-VPD) Surveillance Network reported data from 2014 to 2019, contributing to the estimates of the disease burden and serotypes of pediatric meningitis caused by Streptococcus pneumoniae, Haemophilus influenzae and Neisseria meningitidis.

The CAPACITI Decision-Support Tool for National Immunization Programs.

OBJECTIVES: Immunization programs in low-income and middle-income countries (LMICs) are faced with an ever-growing number of vaccines of public health importance recommended by the World Health Organization, while also financing a greater proportion of the program through domestic resources. More than ever, national immunization programs must be equipped to contextualize global guidance and make choices that are best suited to their setting. The CAPACITI decision-support tool has been developed in collaboration with national immunization program decision makers in LMICs to structure and document an evidence-based, context-specific process for prioritizing or selecting among multiple vaccination products, services, or strategies. METHODS: The CAPACITI decision-support tool is based on multi-criteria decision analysis, as a structured way to incorporate multiple sources of evidence and stakeholder perspectives. The tool has been developed iteratively in consultation with 12 countries across Africa, Asia, and the Americas. RESULTS: The tool is flexible to existing country processes and can follow any type of multi-criteria decision analysis or a hybrid approach. It is structured into 5 sections: decision question, criteria for decision making, evidence assessment, appraisal, and recommendation. The Excel-based tool guides the user through the steps and document discussions in a transparent manner, with an emphasis on stakeholder engagement and country ownership. CONCLUSIONS: Pilot countries valued the CAPACITI decision-support tool as a means to consider multiple criteria and stakeholder perspectives and to evaluate trade-offs and the impact of data quality. With use, it is expected that LMICs will tailor steps to their context and streamline the tool for decision making.

Evaluation of Tween 80 incorporated media to increase pathogen isolation from peritoneal fluid of CAPD patients at Hospital Kuala Lumpur.

INTRODUCTION: Continuous ambulatory peritoneal dialysis (CAPD)-associated peritonitis remains a major complication in patients on CAPD leading to increased morbidity and mortality. Successful therapy of peritonitis is highly dependent on a positive microbiological culture because narrow spectrum antibiotics are essential to efficiently combat infection. Therefore, this study evaluated the performance of Tween 80 containing media at three different concentrations (0.1%, 1.0% and 2.0%) to increase the pathogen yield from peritoneal fluid in comparison with the standard culture media. MATERIALS AND METHODS: Peritoneal fluid samples (n=121) obtained from CAPD patients suspected of peritonitis at Hospital Kuala Lumpur were analysed macroscopically and microscopically prior to culture. All samples were cultured on seven different culture media, including sheep blood agar, MacConkey agar, Sabouraud dextrose agar, brain heart infusion agar and Tween 80 incorporated blood agar. All plates were incubated at an optimum temperature up to 48 hours. RESULTS AND CONCLUSION: Among all the culture media investigated, 0.1% to 2.0% Tween 80 incorporated blood agar yielded the highest positive culture (23/121) in comparison with all other standard media, thus lowering the negative culture rate among CAPD patients. Statistical analysis by Chi Square revealed significant differences (p <0.001) between the three concentrations of Tween 80 tested in this study. Among the three different concentrations of Tween 80 optimised in this study, blood agar containing 0.1% Tween 80 generated the best results, achieved by optimum growth of all Gram-positive organisms, Gram-negative organisms and yeast cells simultaneously. Using a small amount of detergent at low cost significantly increased the pathogen yield during CAPD-associated peritonitis.

Diagnosis of neonatal meningitis: is it time to use polymerase chain reaction?

Group B Streptococcus (GBS) is a predominant causative pathogen of neonatal meningitis that is associated with a high rate of mortality and morbidity. The establishment of antenatal screening and intrapartum chemoprophylaxis has led to a significant reduction in the incidence rate of invasive GBS disease in developed countries. However, these strategies are not routinely practiced in most developing countries. To ensure good recovery of infants affected with GBS, a prompt diagnostic strategy and appropriate therapy are essential. We highlight here the case of a preterm male infant diagnosed with early-onset of GBS meningitis diagnosed by using polymerase chain reaction (PCR) method on the cerebrospinal fluid (CSF) of the infant. Initially the pathogen was not isolated in both blood and CSF cultures as sampling was performed after the administration of antibiotics. Hence, PCR was a crucial diagnostic test in facilitating the detection of the pathogen in CSF. We believe that PCR is a potentially fast and precise diagnostic method for infection in a newborn.

Global impact of rotavirus vaccine introduction on rotavirus hospitalisations among children under 5 years of age, 2008-16: findings from the Global Rotavirus Surveillance Network.

BACKGROUND: Rotavirus vaccine use in national immunisation programmes has led to declines in hospital admissions for rotavirus gastroenteritis among children; however, the global impact of rotavirus vaccine introduction has not been described using primary data. We describe the impact of rotavirus vaccine introduction on admissions for acute rotavirus gastroenteritis in primarily low-income and middle-income countries, using 9 years of data from the WHO-coordinated Global Rotavirus Surveillance Network (GRSN). METHODS: Between Jan 1, 2008, and Dec 31, 2016, children younger than 5 years of age who were admitted to hospital with acute gastroenteritis were prospectively enrolled in GRSN sites. We included sites that enrolled children and collected stool specimens monthly and tested at least 100 specimens annually in the impact analysis, with a separate analysis taking into account site continuity. We compared proportions of acute gastroenteritis cases positive for rotavirus in the pre-vaccine and post-vaccine periods and calculated mean proportion changes for WHO regions, with 95% CIs; these findings were then compared with interrupted time series analyses. We did further sensitivity analyses to account for rotavirus vaccination coverage levels and sites that collected specimens for at least 11 months per year and tested at least 80 specimens per year. We also analysed the age distribution of rotavirus-positive cases before and after vaccine introduction. FINDINGS: 403 140 children younger than 5 years of age admitted to hospital with acute gastroenteritis from 349 sites in 82 countries were enrolled over the study period, of whom 132 736 (32·9%) were positive for rotavirus. We included 305 789 children from 198 sites in 69 countries in the impact analysis. In countries that had not introduced rotavirus vaccine in their national immunisation programmes, rotavirus was detected in 38·0% (95% CI 4·8-73·4) of admissions for acute gastroenteritis annually whereas in those that have introduced the vaccine, rotavirus was detected in 23·0% (0·7-57·7) of admissions for acute gastroenteritis, showing a 39·6% (35·4-43·8) relative decline following introduction. Interrupted time series analyses confirmed these findings. Reductions by WHO regions ranged from 26·4% (15·0-37·8) in the Eastern Mediterranean Region to 55·2% (43·0-67·4) in the European Region and were sustained in nine countries (contributing up to 31 sites) for 6-10 years. The age distribution of children with rotavirus gastroenteritis shifted towards older children after rotavirus vaccine introduction. INTERPRETATION: A significant and sustained reduction in the proportion of hospital admissions for acute gastroenteritis due to rotavirus was seen among children younger than 5 years in GRSN sites following rotavirus vaccine introduction. These findings highlight the need to incorporate rotavirus vaccines into immunisation programmes in countries that have not yet introduced them and underline the importance of high-quality surveillance. FUNDING: The GRSN receives funding from Gavi, the Vaccine Alliance and the US Centers for Disease Control and Prevention. No specific funding was provided for this Article.

Vitamin D deficiency in patients with tuberculous meningitis and its relationship with treatment outcome.

SETTING: Data on vitamin D deficiency in tuberculous meningitis (TBM) and its relationship with treatment outcomes are limited. Some of the beneficial effects of vitamin D might be mediated through interleukin-1ß (IL-1ß). OBJECTIVE: To assess the frequency of vitamin D deficiency among TBM patients, its association with treatment outcomes and correlation between vitamin D and IL-1ß levels in cerebrospinal fluid (CSF). DESIGN: We prospectively studied a consecutive sample of human immunodeficiency virus-negative patients with TBM treated at a hospital in southern India. We defined good outcome as survival without severe neurological disability. Serum total 25-hydroxy vitamin D (25[OH]D) and IL-1ß levels in CSF were estimated on pretreatment samples. RESULTS: We studied 40 patients with TBM; 22 (55%) patients had stage 3 disease. Treatment outcome was poor in 21 (53%) patients: 15 (38%) patients died and 6 (15%) had severe neurological disability. The overall mean serum 25(OH)D level was 32.30 ± 16.38 ng/ml. Ten (25%) patients had vitamin D deficiency (<20 ng/ml), and 12 (30%) patients had vitamin D insufficiency (20-30 ng/ml). However, pretreatment serum 25(OH)D levels did not differ significantly by outcome (good vs. poor outcome: 28.30 ± 14.96 vs. 35.92 ± 17.11 ng/ml, P = 0.141). Moreover, IL-1ß levels in CSF did not correlate with serum 25(OH)D levels (Spearman's ρ 0.083, P = 0.609). CONCLUSION: Vitamin D deficiency/insufficiency is common among patients with TBM. However, serum 25(OH)D levels are not associated with IL-1ß levels in CSF or treatment outcome.

FLOOZY of petunia is a flavin mono-oxygenase-like protein required for the specification of leaf and flower architecture.

The mechanisms that determine the relative positions of floral organs, and thereby their numbers, is a poorly understood aspect of flower development. We isolated a petunia mutant, floozy (fzy), in which the formation of floral organ primordia in the outermost three floral whorls and one of the two bracts at the base of the flower is blocked at an early stage. In addition, fzy mutants fail to generate secondary veins in leaves and bracts and display a decreased apical dominance in the inflorescence. FZY encodes an enzyme with homology to flavin mono-oxygenases and appears to be the ortholog of YUCCA genes of Arabidopsis. FZY is expressed in young leafs and bracts and in developing flowers. In young floral meristems FZY is expressed in the center of the meristem dome and, later, expression becomes localized on the flanks of the initiating petal and stamen primordia and at several sites in maturing anthers and carpels. These findings indicate that FZY is involved in synthesizing a signaling compound that is required for floral organ initiation and specification of the vascularization pattern in leaves. Although fzy mutants contain normal auxin levels, ectopic expression of FZY results in excessive auxin accumulation, suggesting that the signaling compound is auxin.

Modulation of cell proliferation in the embryonic retina of zebrafish (Danio rerio).

We describe light-microscopically the development of the embryonic zebrafish eye with particular attention to cell number, cell proliferation, and cell death. The period from 16 to 36 hr post fertilization (hpf) comprises two phases; during the first (16-27 hpf) the optic vesicle becomes the eye cup, and during the second (27-36 hpf) the eye cup begins to differentiate into the neural retina and pigmented epithelium. All cells in the eye primordium are proliferative prior to 28 hpf, and the length of the cell cycle has been estimated to be 10 hr at 24-28 hpf (Nawrocki, 1985). Our cell counts are consistent with that estimate at that age, but not at earlier ages. A 10-hr cell cycle predicts that the cell number should increase by 7% per hr, but during 16-24 hpf the cell number increased by only 1.5% per hr. Despite the low rate of increase, all cells labeled with bromo-deoxyuridine, so all were proliferative. We considered three possible explanations for the nearly-constant cell number in the first phase: proliferation balanced by cell emigration from the eye, proliferation balanced by cell death, and low proliferation caused by a transient prolongation of the cell cycle. We excluded the first two, and found direct support for the third. Previous examinations of the cell cycle length in vertebrate central nervous system have concluded that it increases monotonically, in contrast to the modulation that we have shown. Modulation of the cell cycle length is well-known in flies, but it is generally effected by a prolonged arrest at one phase, in contrast to the general deceleration that we have shown.

The morphogenesis of the zebrafish eye, including a fate map of the optic vesicle.

We have examined the morphogenesis of the zebrafish eye, from the flat optic vesicle at 16 hours post fertilization (hpf) to the functional hemispheric eye at 72 hpf. We have produced three-dimensional reconstructions from semithin sections, measured volumes and areas, and produced a fate map by labeling clusters of cells at 14-15 hpf and finding them in the 24 hpf eye cup. Both volume and area increased sevenfold, with different schedules. Initially (16-33 hpf), area increased but volume remained constant; later (33-72 hpf) both increased. When the volume remained constant, the presumptive pigmented epithelium (PE) shrank and the presumptive neural retina (NR) enlarged. The fate map revealed that during 14-24 hpf cells changed layers, moving from the PE into the NR, probably through involution around the margin of the eye. The transformation of the flat epithelial layers of the vesicle into their cup-shaped counterparts in the eye was also accompanied by cellular rearrangements; most cells in a cluster labeled in the vesicle remained neighbors in the eye cup, but occasionally they were separated widely. This description of normal zebrafish eye development provides explanations for some mutant phenotypes and for the effects of altered retinoic acid.