Epidemiological Surveillance: Genetic Diversity of Rotavirus Group A in the Pearl River Delta, Guangdong, China in 2019

Objective This study aimed to understand the epidemic status and phylogenetic relationships of rotavirus group A(RVA)in the Pearl River Delta region of Guangdong Province,China.Methods This study included individuals aged 28 days–85 years.A total of 706 stool samples from patients with acute gastroenteritis collected between January 2019 and January 2020 were analyzed for 17 causative pathogens,including RVA,using a Gastrointestinal Pathogen Panel,followed by genotyping,virus isolation,and complete sequencing to assess the genetic diversity of RVA.Results The overall RVA infection rate was 14.59%(103/706),with an irregular epidemiological pattern.The proportion of co-infection with RVA and other pathogens was 39.81%(41/103).Acute gastroenteritis is highly prevalent in young children aged 0–1 year,and RVA is the key pathogen circulating in patients 6–10 months of age with diarrhea.G9P[8](58.25%,60/103)was found to be the predominant genotype in the RVA strains,and the 41 RVA-positive strains that were successfully sequenced belonged to three different RVA genotypes in the phylogenetic analysis.Recombination analysis showed that gene reassortment events,selection pressure,codon usage bias,gene polymorphism,and post-translational modifications(PTMs)occurred in the G9P[8]and G3P[8]strains.Conclusion This study provides molecular evidence of RVA prevalence in the Pearl River Delta region of China,further enriching the existing information on its genetics and evolutionary characteristics and suggesting the emergence of genetic diversity.Strengthening the surveillance of genotypic changes and gene reassortment in RVA strains is essential for further research and a better understanding of strain variations for further vaccine development.

Neonatal erythema multiforme associated with a rotavirus infection:A case report

BACKGROUND Erythema multiforme(EM)is an extremely rare condition in neonates,and studies suggest its association with certain infections and neonatal vaccinations;however,few specific etiological agents have been identified.Rotavirus,a common pathogenic gastrointestinal virus in the neonatal period that is preventable via vaccination,has not been identified as a possible etiology.We report the case of a neonate who was referred for skin lesions presenting as EM,where a meticulous workup identified rotavirus as the sole causative agent.CASE SUMMARY A 14-day-old male infant was admitted to our hospital with a 1-day history of skin lesions.No medical history or medication intake was recorded.Except for the complaint of skin lesions,the caregivers denied any abnormal symptoms.Multiple tests,including routine laboratory evaluations,were performed to identify the cause of skin lesions.Serological tests for Immunoglobulin M for Toxoplasma,Rubella,Cytomegalovirus,Herpes Simplex Virus,and Epstein-Barr virus viral-capsid antigen were all negative.Multiple polymerase chain reaction(PCR)tests for respiratory viruses and bacterial pathogens were negative(including the severe acute respiratory syndrome coronavirus 2).Multiple PCR tests for gastrointestinal viruses and bacterial pathogens demonstrated evidence of rotavirus infection.No growth was reported in the blood and urine cultures.The patient received intravenous fluids for hydration;meanwhile,no other medications were prescribed.The lesions improved rapidly without specific treatment,and full recovery was achieved within a week.CONCLUSION The possibility of rotavirus,a major cause of pediatric gastrointestinal infections,being a trigger for neonatal EM should be considered.

Clinical factors predicting rotavirus diarrhea in children:A crosssectional study from two hospitals

BACKGROUND Rotavirus is still a significant contributing morbidity and mortality in pediatric patients.AIM To look at clinical signs and symptoms and laboratory findings that can predict rotavirus gastroenteritis compared to non-rotavirus gastroenteritis.METHODS This was a cross-sectional study with medical records obtained from December 2015 to December 2019.Inclusion criteria for this study include all hospitalised pediatric patients(0-18 years old)diagnosed with suspected rotavirus diarrhea.The receiver operating curve and Hosmer-Lemeshow test would be used to assess the final prediction findings'calibration(goodness of fit)and discrimination performance.RESULTS This study included 267 participants with 187(70%)rotavirus-diarrhea cases.The patients were primarily male in both rotavirus(65.2%)and non-rotavirus(62.5%)groups.The median age is 1.33 years old(0.08-17.67 years old).Multivariate analysis shows that wet season(OR_(adj)=2.5;95%CI:1.3-4.8,Padj=0.006),length of stay(LOS)≥3 days(OR_(adj)=5.1;95%CI:1.4-4.8,Padj=0.015),presence of abdominal pain(OR_(adj)=3.0;95%CI:1.3-6.8,Padj=0.007),severe dehydration(OR_(adj)=2.9;95%CI:1.1-7.9,Padj=0.034),abnormal white blood cell counts(OR_(adj)=2.8;95%CI:1.3-6.0,Padj=0.006),abnormal random blood glucose(OR_(adj)=2.3;95%CI:1.2-4.4,Padj=0.018)and presence of fecal leukocytes(OR_(adj)=4.1,95%CI:1.7-9.5,Padj=0.001)are predictors of rotavirus diarrhea.The area under the curve for this model is 0.819(95%CI:0.746-0.878,P value<0.001),which shows that this model has good discrimination.CONCLUSION Wet season,LOS≥3 d,presence of abdominal pain,severe dehydration,abnormal white blood cell counts,abnormal random blood glucose,and presence of fecal leukocytes predict rotavirus diarrhea.

Prevalence and Genotypes of Rotavirus A and Human Adenovirus among Hospitalized Children with Acute Gastroenteritis in Fujian, China, 2009-2017

Gastroenteritis is an infectious diarrhea that has been considered as an important cause of hospitalizations and death in children aged < 5 years, particularly in developing countries. Unsanitary water, contaminated food, poor hygiene, and inadequate disposal of waste and feces are all risk factors for gastroenteritis, resulting in the higher incidence in developing countries. Gastroenteritis is generally caused by viral infections, among which rotavirus (RV) infections have been reported to be the most common, especially among young children aged < 5 years with acute gastroenteritis in Asia and Africa[1]. Other viruses associated with acute gastroenteritis include human Adenovirus (HAdV), Norovirus, Sapovirus (SaV), human Astrovirus (HAstV), and Aichi virus. Recent research has reported that adenovirus types 40 and 41, belonging to species F, cause gastroenteritis and were therefore termed as enteric adenoviruses. In addition, non-enteric HAdV species such as A, B, C, and D have been associated with diarrheal.

Lentinan administration alleviates diarrhea of rotavirus-infected weaned pigs via regulating intestinal immunity

Background:Lentinan(LNT)may regulate many important physiological functions of human and animals.This study aimed to verify whether LNT administration could relieve diarrhea via improving gut immunity in rotavirus(RV)-challenged weaned pigs.Methods:Twenty-eight weaned pigs were randomly fed 2 diets containing 0 or 84 mg/kg LNT product for 19 d(n=14).RV infection was executed on d 15.After extracting polysaccharides from LNT product,its major monosaccharides were analyzed.Then,LNT polysaccharide was used to administrate RV-infected IPEC-J2 cells.Results:Dietary LNT supplementation supported normal function of piglets even when infected with RV,as reflected by reduced growth performance loss and diarrhea prevalence,and maintained gut immunity(P<0.05).The polysaccharide was isolated from LNT product,which molecular weight was 5303 Da,and major monosaccharides included glucose,arabinose and galactose.In RV-infected IPEC-J2 cells,this polysaccharide significantly increased cell viability(P<0.05),and significantly increased anti-virus immunity via regulating pattern recognition receptors and host defense peptides(P<0.05).Conclusion:Those results suggest that LNT administration increases the piglets’resistance to RV-induced stress,likely by supporting intestinal immunity.

Expression of NF-kappaB in rotavirus-induced damage to the liver and biliary tract in neonatal mice

BACKGROUND:Biliary atresia, the etiology of which still remains unclear, occurs exclusively in newborns and most are infected with rotavirus. In this study, we aimed to investigate the histopathological patterns of different kinds of rotavirus in the liver and biliary tract of neonatal mice and the expression of NF-κB in the liver and biliary tract of infected mice. METHODS:Twenty-three adult mice (8 were male and 15 female) were divided into 8 breeding pairs, and each pair (1 male and 2 females) was housed in a cage in a laminar flow hood. Newborn mice, 24-48 hours old were randomly divided into A, B and C groups. The A and B groups were respectively inoculated with MMU18006 and SA11 rotavirus through the intraperitoneal route, while group C as blank control was only inoculated with culture medium. The liver was dissected after 5, 10, 15, 21 and 28 days; the weight of each mouse and the histopathological patterns in the liver were recorded. The expression of NF-κB in the liver and intrahepatic bile ducts was detected by immunohistochemical staining and the expression intensity was analyzed with a GT-2 imaging instrument. RESULTS:The average increase in weight of infected mice was significantly slower than that of the normal control, while the growth rate of group A (injected with MMU18006 rotavirus) was slower than that of group B (SA11 rotavirus). In infected mice, the acute and chronic inflammation of liver and intra-and extra-hepatic bileducts was more significant in group A. Stenosis was found in most intrahepatic bile ducts, and sporadically in extrahepatic bile ducts. The expression of NF-κB in infected mice was dramatically higher than that of the normal control, while the expression in group A was higher than in group B. CONCLUSIONS:Significant damage to the liver and biliary tract of neonatal mice can be induced by inoculating MMU18006 rotavirus through the intraperitoneal route, which is very similar to the pathology of biliary atresia in the newborn human. Similar inoculation with SA11 rotavirus can only result in moderate impairment that disappears quickly. The difference of pathogenicity between the two rotaviruses may depend on their differing capacities to increase the expression of NF-κB in the liver and biliary tract.

Acute effects of rotavirus and malnutrition on intestinal barrier function in neonatal piglets

AIM: To investigate the effect of protein-energy malnutrition on intestinal barrier function during rotavirus enteritis in a piglet model.METHODS: Newborn piglets were allotted at day 4 of age to the following treatments:(1) full-strength formula(FSF)/noninfected;(2) FSF/rotavirus infected;(3) half-strength formula(HSF)/noninfected;or(4) HSF/rotavirus infected.After one day of adjustment to the feeding rates,pigs were infected with rotavirus and acute effects on growth and diarrhea were monitored for 3 d and jejunal samples were collected for Ussingchamber analyses.RESULTS: Piglets that were malnourished or infected had lower body weights on days 2 and 3 post-infection(P < 0.05).Three days post-infection,marked diarrhea and weight loss were accompanied by sharp reductions in villus height(59%) and lactase activity(91%) and increased crypt depth(21%) in infected compared with non-infected pigs(P < 0.05).Malnutrition also increased crypt depth(21%) compared to full-fed piglets.Villus:crypt ratio was reduced(67%) with viral infection.There was a trend for reduction in transepithelial electrical resistance with rotavirus infection and malnutrition(P = 0.1).3H-mannitol flux was significantly increased(50%;P < 0.001) in rotavirus-infected piglets compared to non-infected piglets,but there was no effect of nutritional status.Furthermore,rotavirus infection reduced localization of the tight junction protein,occludin,in the cell membrane and increased localization in the cytosol.CONCLUSION: Overall,malnutrition had no additive effects to rotavirus infection on intestinal barrier function at day 3 post-infection in a neonatal piglet model.

Inflammatory and oxidative stress in rotavirus infection

Rotaviruses are the single leading cause of life-threatening diarrhea affecting children under 5 years of age. Rotavirus entry into the host cell seems to occur by sequential interactions between virion proteins and various cell surface molecules. The entry mechanisms seem to involve the contribution of cellular molecules having binding, chaperoning and oxido-reducing activities. It appears to be that the receptor usage and tropism of rotaviruses is determined by the species, cell line and rotavirus strain. Rotaviruses have evolved functions which can antagonize the host innate immune response, whereas are able to induce endoplasmic reticulum(ER) stress, oxidative stress and inflammatory signaling. A networking between ER stress, inflammation and oxidative stress is suggested, in which release of calcium from the ER increases the generation of mitochondrial reactive oxygen species(ROS) leading to toxic accumulation of ROS within ER and mitochondria. Sustained ER stress potentially stimulates inflammatory response through unfolded protein response pathways. However, the detailed characterization of the molecular mechanisms underpinning these rotavirus-induced stressful conditions is still lacking. The signaling events triggered by host recognition of virusassociated molecular patterns offers an opportunity for the development of novel therapeutic strategies aimed at interfering with rotavirus infection. The use of N-acetylcysteine, non-steroidal anti-inflammatory drugs and PPARγ agonists to inhibit rotavirus infection opens a new way for treating the rotavirus-induced diarrhea and complementing vaccines.

Development of a human rotavirus induced diarrhea model in Chinese mini-pigs

AIM: to establish a new animal model for the research of human rotavirus(HRV) infection, its pathogenesis and immunity and evaluation of potential vaccines.METHODS: 5-d, 30-d and 60-d-old Chinese mini-pigs, Guizhou and bamma, were inoculated with a single oral dose of attenuated strain Wa, G1, G3 of HRV, and PbS(control), respectively, and fecal samples of pigs from 0 to 7 d post infection(DPI) were collected individually. Enzyme linked immunosorbent assay was used to detect HRV antigen in feces. the HRV was tested by real-time PCR(Rt-PCR). the sections of the intestinal tissue were stained with hematoxylin and eosin to observe the morphologic variation by microscopy. Immunofluorescence was used to determine the HRV in intestinal tissue. HRV particles in cells of the ileum were observed by electron micrography.RESULTS: When inoculated with HRV, mini-pigs younger than 30 d developed diarrhea in an agedependent manner and shed HRV antigen of the sameinoculum, as demonstrated by Rt- PCR.Histopathological changes were observed in HRV inoculated mini-pigs including small intestinal cell tumefaction and necrosis. HRV that was distributed in the small intestine was restricted to the top part of the villi on the internal wall of the ileum, which was observed by immunofluorescence and transmission electron microscopy. Virus particles were observed in Golgi like follicles in HRV-infected neonatal minipigs. Guizhou mini-pigs were more sensitive to HRV than bamma with respect to RV antigen shedding and clinical diarrhea.CONCLUSION: these results indicate that we have established a mini-pig model of HRV induced diarrhea. Our findings are useful for the understanding of the pathogenic mechanisms of HRV infection.

Epidemiological and Clinical Features of Rotavirus and Adenovirus Related Gastroenteritis in Beijing:A Retrospective Case-control Study in Pediatric Patients

Rotavirus(RV)is the most common cause of viral gastroenteritis among children younger than 5-yearold worldwide.RV has nine groups(Group A to I)and Group A(RVA)is the main cause of severe gastroenteritis disease in children.Human adenovirus(HAdV)consists of 7 species(HAdV-A through HAdV-G)including over 70 serotypes,and group F serotypes 40 and 41 are related to gastroenteritis[1].There were reports that revealed co-infection of AdV with RVA and Norovirus[2].Here we report an epidemiological and clinical analysis of RVA and AdV infection through a single-centered retrospective case-control study.

Neonatal rhesus monkeys as an animal model for rotavirus infection

AIM To establish a rotavirus(RV)-induced diarrhea model using RV SA11 in neonatal rhesus monkeys for the study of the pathogenic and immune mechanisms of RV infection and evaluation of candidate vaccines.METHODS Neonatal rhesus monkeys with an average age of 15-20 d and an average weight of 500 g ± 150 g received intragastric administration of varying doses of SA11 RV( 107 PFUs/mL, 106 PFUs/mL, or 105 PFUs/mL, 10 mL/animal) to determine whether the SA11 strain can effectively infect these animals by observing their clinical symptoms, fecal shedding of virus antigen by ELISA, distribution of RV antigen in the organs by immunofluorescence, variations of viral RNA load in the organs by qRT-PCR, histopathological changes in the small intestine by HE staining, and apoptosis of small intestinal epithelial cells by TUNEL assay.RESULTS The RV monkey model showed typical clinical diarrhea symptoms in the 108 PFUs SA11 group, where we observed diarrhea 1-4 d post infection(dpi) and viral antigen shed in the feces from 1-7 dpi. RV was found in jejunal epithelial cells. We observed a viral load of approximately 5.85 × 103 copies per 100 mg in the jejunum at 2 dpi, which was increased to 1.09 × 105 copies per 100 mg at 3 dpi. A relatively high viral load was also seen in mesenteric lymph nodes at 2 dpi and 3 dpi. The following histopathological changes were observed in the small intestine following intragastric administration of SA11 RV: vacuolization, edema, and atrophy. Apoptosis in the jejunal villus epithelium was also detectable at 3 dpi.CONCLUSION Our results indicate that we have successfully established a RV SA11 strain diarrhea model in neonatal rhesus monkeys. Future studies will elucidate the mechanisms underlying the pathogenesis of RV infection, and we will use the model to evaluate the protective effect of candidate vaccines.

Protective effects of sodium butyrate on rotavirus inducing endoplasmic reticulum stress-mediated apoptosis via PERK-eIF2αsignaling pathway in IPEC-J2 cells

Background:Rotavirus(RV)is a major pathogen that causes severe gastroenteritis in infants and young animals.Endoplasmic reticulum(ER)stress and subsequent apoptosis play pivotal role in virus infection.However,the protective mechanisms of intestinal damage caused by RV are poorly defined,especially the molecular pathways related to enterocytes apoptosis.Thus,the aim of this study was to investigate the protective effect and mechanism of sodium butyrate(SB)on RV-induced apoptosis of IPEC-J2 cells.Results:The RV infection led to significant cell apoptosis,increased the expression levels of ER stress(ERS)markers,phosphorylated protein kinase-like ER kinase(PERK),eukaryotic initiation factor 2 alpha(eIF2α),caspase9,and caspase3.Blocking PERK pathway using specific inhibitor GSK subsequently reversed RV-induced cell apoptosis.The SB treatment significantly inhibited RV-induced ERS by decreasing the expression of glucose regulated protein 78(GRP78),PERK,and eIF2α.In addition,SB treatment restrained the ERS-mediated apoptotic pathway,as indicated by downregulation of C/EBP homologous protein(CHOP)mRNA level,as well as decreased cleaved caspase9 and caspase3 protein levels.Furthermore,siRNA-induced GPR109a knockdown significantly suppressed the protective effect of SB on RV-induced cell apoptosis.Conclusions:These results indicate that SB exerts protective effects against RV-induced cell apoptosis through inhibiting ERS mediated apoptosis by regulating PERK-eIF2αsignaling pathway via GPR109a,which provide new ideas for the prevention and control of RV.

Molecular characterization of VP4,VP6,VP7 and NSP4 genes of group B rotavirus strains from outbreaks of gastroenteritis

Objective:To characterize VP4,VP6,VP7 and NSP4 genes of representative GBR strains(NIV- 005625.MV-04622 and NIV-094456) delected as the major eliolngic agenl in the outbreaks of gastroenteritis in western India.Methods:Fecal specimens collected during the outbreaks of gastroenteritis were processed for RNA isolation.RT-PCR using GBR VP4.VP6.VP7 and NSP4 gene specific primers,nucleotide sequencing of the amplicons and phylogenetic analysis of the sequences.Results:Phylogenetic analysis of all of the VP4.VP6.VP7 and NSP4 gene sequences revealed clustering of GBR strains in Indian-Bangladeshi lineage of genotype G2 with 95.8%- 99.4%nucleotide and 97.3%-100.0%amino acid identities.However,all three strains showed the presence of unique amino acid substitutions in the VP4 protein suggesting alteration in the antigenicity of outbreak strains of GBR.The VP8* and VP5* regions of VP4 proteins showed respectively 0.5%-6.3%and 0.2%-1.1%amino acid divergence from human GBR strains of Indian-Bangladeshi lineage.Conclusions:These data confirm the reported variability of VP8* region and suggest the possible role of this region in the perpetuation of GBR infections in the environment.This is the first study to document the phylogenetic relationship of VP4,VP6.VP7 and NSP4 genes of GBR strains detected in the outbreaks of gastroenteritis from India with the CBR strains from other parts of world.

Isolation and characterization of a new candidate human inactivated rotavirus vaccine strain from hospitalized children in Yunnan,China:2010-2013

AIM To determine the distribution of rotavirus VP7 gene in hospitalized children in Yunnan, China. METHODS A total of 366 stool specimens were collected from hospitalized children in hospitals in Yunnan Province from September 2010 to December 2013. The genomic RNA electropherotypes and the G genotypes of the rotaviruses were determined. A phylogenetic analysis of the VP7 gene was performed. Rotavirus isolation was performed, and characterized by plaque, minimum essential medium, and all genes sequence analysis. Quantification of antibodies for inactivated vaccine prepared with ZTR-68 was examined by enzyme-linked immunosorbent assay and microneutralization assay.RESULTS Group A human rotavirus was detected in 177 of 366(48.4%) stool samples using a colloidal gold device assay. The temporal distribution of rotavirus cases showed significant correlation with the mean air temperature. Rotaviruses were isolated from 13% of the rotavirus-positive samples. The predominant genotype was G1(43.5%), followed by G3(21.7%), G9(17.4%), G2(4.3%), G4(8.7%), and mixed(4.3%) among a total of 23 rotavirus isolates. A rotavirus strain was isolated from a rotavirus-positive stool sample of a 4-month-old child in The First People's Hospital of Zhaotong(2010) for use as a candidate human inactivated rotavirus vaccine strain and for further research, and was designated ZTR-68. The genotype of 11 gene segments of strain ZTR-68(RVA/Human-wt/CHN/ZTR-68/2010/G1P[8]) was characterized. The genotype constellation of strain ZTR-68 was identified as G1-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1. The VP7 and VP4 genotypes of strain ZTR-68 were similar to Wa-like strains.CONCLUSIONS A high prevalence of the G1, G2, and G3 genotypes was detected from 2010 to 2012. However, a dominant prevalence of the G9 genotype was identified as the cause of gastroenteritis in children in Yunnan, China, in 2013. A candidate human inactivated rotavirus vaccine strain, designated ZTR-68 was isolated, characterized, and showed immunogenicity. Our data will be useful for the future formulation and development of a vaccine in China.

Rapid genotyping of human rotavirus using SYBR green real-time reverse transcription-polymerase chain reaction with melting curve analysis

AIM: To develop a real-time reverse transcriptionpolymerase chain reaction(RT-PCR) assay to genotype rotavirus(G and P) in Alberta from January 2012 to June 2013. METHODS: We developed and validated a different approach to perform rotavirus G and P genotyping using a two-step SYBR green RT-PCR(rt-g PCR) by selecting genotype-specific primers of published conventional RT nested PCR(cn RT-PCR) assay and optimizing the amplification conditions. c DNA was first synthesized from total RNA with Super Script? Ⅱ reverse transcriptase kit followed by amplication step using monoplex SYBR green real-time PCR. After the PCR reaction, melting curve analysis was used to determine specific genotype. Sixteen samples previously genotyped using cn RT-PCR were tested using the new assay and the genotyping results were compared as sensitivity analysis. Assay specificity was evaluated by testing other gastroenteritis viruses with the new assay. The amplicon size of each available genotype was determined by gelelectrophoresis and DNA sequences were obtained using Sanger-sequencing method. After validation and optimization, the new assay was used to genotype 122 pediatric clinical stool samples previously tested positive for rotavirus using electron microscopy between January2012 and June 2013.RESULTS: The new rt-g PCR assay was validated and optimized. The assay detected G1 to G4, G9, G12 and P[4] and P[8] that were available as positive controls in our laboratory. A single and clear peak of melting curve was generated for each of specific G and P genotypes with a Tm ranging from 80 ℃ to 82 ℃. The sensitivity of rt-g PCR was comparable to cn RT-PCR with 100% correlation of the 16 samples with known G and P genotypes. No cross reaction was found with other gastroenteritis viruses. Using the new rt-g PCR assay, genotypes were obtained for 121 of the 122 pediatric clinical samples tested positive for rotavirus: G1P[8](42.6%), G2P[4](4.9%), G3P[8](10.7%), G9P[8](10.7%), G9P[4](6.6%), G12P[8](23.0%), and unknown GP[8](0.8%). For the first time, G12 rotavirus strains were found in Alberta and G12 was the second most common genotype during the study period. Gel electrophoresis of all the genotypes showed expected amplicon size for each genotype. The sequence data of the two G12 samples along with other genotypes were blasted in NCBI BLAST or analyzed with Rota C Genotyping tool(http://rotac.regatools.be/). All genotyping results were confirmed to be correct.CONCLUSION: rt-g PCR is a useful tool for the genotyping and characterization of rotavirus. Monitoring of rotavirus genotypes is important for the identification of emerging strains and ongoing evaluation of rotavirus vaccination programs.

Prevalence of adenovirus and rotavirus infection in immunocompromised patients with acute gastroenteritis in Portugal

AIM: To characterize the prevalence of rotavirus(RV) and adenovirus(Ad V) infections in immunocompromised patients with acute gastroenteritis. METHODS: The presence of RV and Ad V(serotypes 40 and 41) was evaluated in 509 stool samples obtained between January 2009 and December 2010 from 200 immunocompromised patients(83 females and 117 males; median age 21 years old, range 0-72. The diagnosis of infection was performed as a routine procedure and the presence of RV and Ad V(serotypes 40 and 41) was determined by immunochromatography using the RIDA&#174; Quick Rota-Adeno-Kombi kit(r-Biopharm, Darmstadt, Germany). The data analysis and description of seasonal frequencies were performed using computer software IBM&#174; SPSS&#174;(Statistical Package for Social Sciences) Statistics version 20.0 for Mac. The frequencies of infection were compared into different age and gender groups by χ2 test.RESULTS: The study revealed 12.4% Ad V positive samples and 0.8% RV positive samples, which correspond to a prevalence of 6.5% and 1.5%, respectively. Ad V was more frequent between October 2009 and April 2010, while RV was identified in April 2010 and July 2010. The stool analysis revealed that from the 509 samples, 63(12.4%) were positive for Ad V and 4(0.8%) positive for RV, which by resuming the informationof each patient, lead to an overall prevalence of Ad V and RV of 6.5%(13/200 patients) and 1.5%(3/200 patients), respectively. The stratification of the analysis regarding age groups showed a tendency to an increased prevalence of infection in paediatric patients between 0-10 years old. Considering the seasonal distribution of these infections, our study revealed that Ad V infection was more frequent between October 2009 and April 2010, while RV infection was characterized by two distinct peaks(April 2010 and July 2010). CONCLUSION: The overall prevalence of Ad V and RV infection in immunocompromised patients with acute gastroenteritis was 8% and Ad V was the most prevalent agent.

Successes, Challenges and Lessons Learned in Accelerating Introduction of Rotavirus Immunisation in Zambia

Introduction: Under five mortality in Zambia is unacceptably high and diarrhoea is the third leading contributor. The Programme for Awareness and Elimination of Diarrhoea (PAED) sought to support the government to accelerate the introduction of new vaccines, including the pneumococcal, second dose measles and rotavirus vaccines in Zambia. Here we present our approach, progress and lessons learned in two years of the programme. Stakeholder Engagement: Definite commitment and buy-in and sign off by the MOH were fundamental prerequisites. National and international stakeholders including the Inter Agency Coordinating Committee (ICC), GAVI Alliance, WHO, University Teaching Hospital, Paediatrics Association of Zambia, and UNICEF were engaged for stakeholder buy-in and integration. Progress made: Following successful integration, PAED was officially launched in January 2012. Preparatory work done included: Introduction and acceptance of the PAED agenda in ICC, new vaccines proposal to GAVI, resource mobilisation, Effective Vaccine Management implementation, national cold chain scale-up strategy, vaccine orientation and adapted data collection tools, health worker training, step-wise vaccine introduction to Lusaka province districts and finally national roll-out of the rotavirus vaccine immunisation. Between January 2011 and November 2013, over 270,000 vaccine doses were distributed in Lusaka province. When 94,500 infants were fully immunised, adequate preparations had been made to facilitate national launch of rotavirus immunisations countrywide on 27th November 2013. Discussion: The PAED model was successful at resource mobilization;it has demonstrated how private sector can contribute to new vaccine introduction. Lessons learned from this model can be replicated in other countries with similar need and constraints.

Evaluation of Hyperimmune Hen Egg Yolk Derived Anti-Human Rotavirus Antibodies (Anti-HRVIgY) against Rotavirus Infection

Oral delivery of specific IgY has been reported to be beneficial against rotavirus infection. However, the production of IgYs against globally prevalent human rotavirus (HRV) serotypes and their evaluation detailing the influence on the virological/histopathological consequences have not been reported to date. In the present study, anti-HRVIgY was generated in the eggs of specific pathogen free hens immunized with HRV serotypes G1 - G4 and G9 independently. Purified anti-HRVIgY preparations were tested to determine the ELISA and neutralizing antibody titers respectively in an indirect ELISA and cell culture based neutralization assay. Efficacy of pre and post infection treatment of anti-HRV-3IgY was assessed in an infant BALB/c mouse model of human rotavirus infection by monitoring percent diarrhea, severity and duration of diarrhea, intestinal viral load and histopathology. High (1:64000 - 1:512000) titered anti-HRVIgYs were obtained from the egg yolk of immunized hens with peak titer value (1:256000/1:512000) at 40 - 60 day of immunization. In-vitro, each of the anti-HRVIgY preparations showed the presence of multiserotypic neutralizing activity with high (1:1600 - ≥1:6400) homologous and low (≤1:50 - 1:800) heterologous titers. However, anti-HRV-3IgY neutralized all of the serotypes tested in the study indicating broader in-vitro neutralizing activity. In mice, post exposure treatment with anti-HRV-3IgY significantly reduced the extent of diarrhea and intestinal virus load and inhibited histopathological changes whereas pre exposure anti-HRV-3IgY treatment imparted immediate protection from development of rotavirus gastroenteritis. Thus, the anti-HRVIgY administered orally decreased morbidity and disease incidences in mice suggesting its potential implication in prophylactic and therapeutic usage in human to achieve reduction in rotavirus disease burden.

The Incidence of Rotavirus and Adenovirus Infections among Children with Diarrhea in Sulaimani Province, Iraq

Human rotavirus and adenovirus infections are major causes of acute outbreaks and sporadic cases of gastroenteritis, occurring primarily among children less than 5 years of age. Little is known about the epidemiology of rotavirus and enteric adenovirus infections in Sulaimani and Iraq. The aim of this study was to determine the incidence and clinical significance of rotavirus and enteric adenovirus gastroenteritis and also to determine possible risk factors for rotavirus and adenovirus gastroenteritis using new simple rapid screening test (VIKIA ROTA AND ADENO). This is a qualitative test based on the immunochromatography technique. In the study, one hundred children less than 5 years of age with acute gastroenteritis admitted to Sulaimani Paediatric Hospital were studied. Rotavirus was identified in 22% of the children, adenovirus was identified in 3% of the children, and mixed rotavirus and adenovirus was identified in 2% of the children. All positive cases were younger than 2 years of age. The findings show that rotavirus is most commonly detected. However, there were no significant associations between rotavirus and adenovirus and gender, type of feeding, geographical distribution, the source of drinking water, and the past history of admission to hospital.

Impact of rotavirus vaccine on acute gastroenteritis emergency department visits and hospitalizations in a highly-vaccinated urban cohort

Background: Rotavirus vaccines (RVV) have significantly reduced rotavirus disease in children over the past 4 years in the United States. In this study, we describe the impact of RVV in preventing acute gastroenteritis (AGE) hospital encounters in a highly-vaccinated urban pediatric network during the 2007 and 2008 rotavirus seasons. Methods: We used 5 urban practices from a practice-based network to conduct a retrospective cohort study comparing the numbers of AGE emergency department (ED) visits and hospitalizations in RVV-immunized (exposed) and non-immunized (unexposed) children during the first 2 full seasons following RVV introduction. We determined incident rate ratios (IRR), using Poisson regression, and vaccine effectiveness for each outcome. Results: The 2007 and 2008 cohorts were analyzed separately. 62% of the 2007 cohort was vaccinated and 88% of the 2008 cohort. AGE hospitalizations were significantly reduced among RVV-immunized children from the 2007 cohort in the 2008 season with vaccine effectiveness of 67%. Sub-analysis of this cohort by age revealed that RVV was most protective against hospitalizations in the youngest age group (IRR = 0.21, 95% CI (0.06, 0.82). A trend toward protection against hospitalization was detected for both cohorts in the first season following immunization that did not reach a statistically significant level. For AGE ED visits, no significant difference was seen between RVV-immunized and non-immunized children in either cohort, although there was a trend toward protection (IRR’s: 0.67 - 0.7). Conclusions: RVV was highly effective in preventing AGE hospitalizations for a subset of our cohort in 2008. Given reports of RVV effectiveness, we hypothesize that herd immunity is responsible for the inability to detect a significant difference between RVV-immunized and non-immunized children in our highly- vaccinated cohort.