RESUMO
The morbidity and mortality resulting from acute gastroenteritis and associated chronic sequelae represent a substantial burden on health care systems worldwide. Few studies have investigated changes in the gut microbiome following an episode of acute gastroenteritis. By using nondirected 16S rRNA gene amplicon sequencing, the fecal microbiota of 475 patients with acute gastroenteritis was examined. Patient age was correlated with the overall microbial composition, with a decrease in the abundance of Faecalibacterium being observed in older patients. We observed the emergence of a potential Escherichia-Shigella-dominated enterotype in a subset of patients, and this enterotype was predicted to be more proinflammatory than the other common enterotypes, with the latter being dominated by Bacteroides or Faecalibacterium The increased abundance of Escherichia-Shigella did not appear to be associated with infection with an agent of a similar sequence similarity. Stool color and consistency were associated with the diversity and composition of the microbiome, with deviations from the norm (not brown and solid) showing increases in the abundances of bacteria such as Escherichia-Shigella and Veillonella Analysis of enriched outliers within the data identified a range of genera previously associated with gastrointestinal diseases, including Treponema, Proteus, Capnocytophaga, Arcobacter, Campylobacter, Haemophilus, Aeromonas, and Pseudomonas Our data represent the first in-depth analysis of gut microbiota in acute gastroenteritis. Phenotypic changes in stool color and consistency were associated with specific changes in the microbiota. Enriched bacterial taxa were detected in cases where no causative agent was identified by using routine diagnostic tests, suggesting that in the future, microbiome analyses may be utilized to improve diagnostics.
Assuntos
Bactérias/isolamento & purificação , Gastroenterite/etiologia , Microbioma Gastrointestinal , Doença Aguda , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Bactérias/classificação , Criança , Pré-Escolar , Fezes , Gastroenterite/microbiologia , Humanos , Lactente , Pessoa de Meia-Idade , Adulto JovemRESUMO
Norovirus is estimated to cause 677 million annual cases of gastroenteritis worldwide, resulting in 210,000 deaths. As viral gastroenteritis is generally self-limiting, clinical samples for epidemiological studies only partially represent circulating noroviruses in the population and is biased towards severe symptomatic cases. As infected individuals from both symptomatic and asymptomatic cases shed viruses into the sewerage system at a high concentration, waste water samples are useful for the molecular epidemiological analysis of norovirus genotypes at a population level. Using Illumina MiSeq and Sanger sequencing, we surveyed circulating norovirus within Australia and New Zealand, from July 2014 to December 2016. Importantly, norovirus genomic diversity during 2016 was compared between clinical and waste water samples to identify potential pandemic variants, novel recombinant viruses and the timing of their emergence. Although the GII.4 Sydney 2012 variant was prominent in 2014 and 2015, its prevalence significantly decreased in both clinical and waste water samples over 2016. This was concomitant with the emergence of multiple norovirus strains, including twoGII.4 Sydney 2012 recombinant viruses, GII.P4 New Orleans 2009/GII.4 Sydney 2012 and GII.P16/GII.4 Sydney 2012, along with three other emerging strains GII.17, GII.P12/GII.3 and GII.P16/GII.2. This is unusual, as a single GII.4 pandemic variant is generally responsible for 65-80% of all human norovirus infections at any one time and predominates until it is replaced by a new pandemic variant. In sumary, this study demonstrates the combined use of clinical and wastewater samples provides a more complete picture of norovirus circulating within the population.
Assuntos
Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/virologia , Norovirus/genética , Norovirus/isolamento & purificação , Águas Residuárias/virologia , Infecções por Caliciviridae/diagnóstico , Doenças Transmissíveis Emergentes/diagnóstico , Doenças Transmissíveis Emergentes/epidemiologia , Doenças Transmissíveis Emergentes/transmissão , Doenças Transmissíveis Emergentes/virologia , Gastroenterite/epidemiologia , Gastroenterite/virologia , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Norovirus/classificação , Pandemias/prevenção & controle , Filogenia , Prevalência , RNA Viral/genéticaRESUMO
BACKGROUND: Norovirus (NoV) is the major cause of acute gastroenteritis across all age groups. In particular, variants of genogroup II, genotype 4 (GII.4) have been associated with epidemics globally, occurring approximately every three years. The pandemic GII.4 variant, Sydney 2012, was first reported in early 2012 and soon became the predominant circulating NoV strain globally. Despite its broad impact, both clinically and economically, our understanding of the fundamental diversity and mechanisms by which new NoV strains emerge remains limited. In this study, we describe the molecular epidemiological trends of NoV-associated acute gastroenteritis in Australia and New Zealand between January 2013 and June 2014. METHODOLOGY: Overall, 647 NoV-positive clinical faecal samples from 409 outbreaks and 238 unlinked cases of acute gastroenteritis were examined by RT-PCR and sequencing. Phylogenetic analysis was then performed to identify NoV capsid genotypes and to establish the temporal dominance of circulating pandemic GII.4 variants. Recombinant viruses were also identified based on analysis of the ORF1/2 overlapping region. FINDINGS: Peaks in NoV activity were observed, however the timing of these epidemics varied between different regions. Overall, GII.4 NoVs were the dominant cause of both outbreaks and cases of NoV-associated acute gastroenteritis (63.1%, n = 408/647), with Sydney 2012 being the most common GII.4 variant identified (98.8%, n = 403/408). Of the 409 reported NoV outbreaks, aged-care facilities were the most common setting in both Western Australia (87%, n = 20/23) and New Zealand (58.1%, n = 200/344) while most of the NoV outbreaks were reported from hospitals (38%, n = 16/42) in New South Wales, Australia. An analysis of a subset of non-GII.4 viruses from all locations (125/239) showed the majority (56.8%, n = 71/125) were inter-genotype recombinants. These recombinants were surprisingly diverse and could be classified into 18 distinct recombinant types, with GII.P16/GII.13 (24% of recombinants) the most common. CONCLUSION: This study revealed that following its emergence in 2012, GII.4 Sydney 2012 variant continued to be the predominant cause of NoV-associated acute gastroenteritis in Australia and New Zealand between 2013 and 2014.
Assuntos
Infecções por Caliciviridae/epidemiologia , Infecções por Caliciviridae/virologia , Gastroenterite/epidemiologia , Gastroenterite/virologia , Norovirus , Variação Antigênica , Austrália/epidemiologia , Surtos de Doenças , Monitoramento Epidemiológico , Genótipo , Humanos , Epidemiologia Molecular , Mutação , Nova Zelândia/epidemiologia , Norovirus/classificação , Norovirus/genética , Norovirus/imunologia , Filogenia , RNA Viral/genética , Recombinação Genética , Proteínas Estruturais Virais/genética , Proteínas Estruturais Virais/imunologiaRESUMO
Given that Campylobacter jejuni is recognized as the most common cause of bacterial gastroenteritis worldwide, recent findings showing comparable levels of Campylobacter concisus in patients with gastroenteritis would suggest that this bacterium is clinically important. The prevalence and abundance of Campylobacter concisus in stool samples collected from patients with acute gastroenteritis was examined using quantitative real-time PCR. The associated virulence determinants exotoxin 9 and zonula occludens toxin DNA were detected for Campylobacter concisus-infected samples using real-time PCR. Campylobacter concisus was detected at high prevalence in patients with gastroenteritis (49.7â%), higher than that observed for Campylobacter jejuni (â¼5â%). The levels of Campylobacter concisus were putatively classified into clinically relevant and potentially transient subgroups based on a threshold developed using Campylobacter jejuni levels, as the highly sensitive real-time PCR probably detected transient passage of the bacterium from the oral cavity. A total of 18â% of patients were found to have clinically relevant levels of Campylobacter concisus, a significant number of which also had high levels of one of the virulence determinants. Of these patients, 78â% were found to have no other gastrointestinal pathogen identified in the stool, which strongly suggests a role for Campylobacter concisus in the aetiology of gastroenteritis in these patients. These results emphasize the need for diagnostic laboratories to employ identification protocols for emerging Campylobacter species. Clinical follow-up in patients presenting with high levels of Campylobacter concisus in the intestinal tract is needed, given that it has been associated with more chronic sequelae.
Assuntos
Infecções por Campylobacter/microbiologia , Campylobacter/classificação , Campylobacter/patogenicidade , Gastroenterite/microbiologia , Adolescente , Adulto , Criança , Pré-Escolar , Diarreia/microbiologia , Fezes/microbiologia , Feminino , Regulação Bacteriana da Expressão Gênica/fisiologia , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Virulência , Adulto JovemRESUMO
OBJECTIVE: There is a lack of information about the prevalence of gastrointestinal illnesses in Australia. Current disease surveillance systems capture only a few pathogens. The aim of this study is to describe the epidemiology of infectious gastrointestinal illnesses in Sydney, Australia. METHODS: A retrospective cross-sectional study of patients with gastrointestinal symptoms who visited tertiary public hospitals in Sydney was conducted between 2007 and 2010. Patients with diarrhoea or loose stools with an enteric pathogen detected were identified. Demographic, clinical and potential risk factor data were collected from their medical records. Measures of association, descriptive and inferential statistics were analysed. RESULTS: In total, 1722 patients were included in this study. Campylobacter (22.0%) and Clostridium difficile (19.2%) were the most frequently detected pathogens. Stratified analysis showed that rotavirus (22.4%), norovirus (20.7%) and adenovirus (18.1%) mainly affected children under 5 years; older children (5-12 years) were frequently infected with Campylobacter spp. (29.8%) and non-typhoid Salmonella spp. (24.4%); infections with C. difficile increased with age.Campylobacter and non-typhoid Salmonella spp. showed increased incidence in summer months (December to February), while rotavirus infections peaked in the cooler months (June to November). DISCUSSION: This study revealed that gastrointestinal illness remains a major public health issue in Sydney. Improvement of current disease surveillance and prevention and control measures are required. This study emphasizes the importance of laboratory diagnosis of enteric infections and the need for better clinical data collection to improve management of disease risk factors in the community.
Assuntos
Infecções Bacterianas/epidemiologia , Gastroenterite/epidemiologia , Viroses/epidemiologia , Adolescente , Adulto , Idoso , Infecções Bacterianas/microbiologia , Criança , Pré-Escolar , Estudos Transversais , Diarreia/epidemiologia , Feminino , Gastroenterite/microbiologia , Gastroenterite/virologia , Humanos , Incidência , Lactente , Masculino , Pessoa de Meia-Idade , New South Wales/epidemiologia , Estudos Retrospectivos , Viroses/virologia , Adulto JovemRESUMO
BACKGROUND: Clostridium difficile infection (CDI) is increasingly being found in populations without traditional risk factors. We compared the relative frequency, risk factors, severity, and outcomes of community-onset CDI with hospital-acquired infection. METHODS: This was a retrospective, observational study of CDI at a tertiary hospital campus in Sydney, Australia. Patients aged 15 years and older with a first episode of CDI from January 1 to December 31, 2011 were included. CDI was defined as the presence of diarrhoea with a positive enzyme immunoassay in conjunction with a positive cell cytotoxicity assay, toxin culture, or organism culture. Main outcome measures were onset of infection (hospital or community), risk factors, markers of severity, and outcomes for the two groups. RESULTS: One hundred and twenty-nine cases of CDI infection were identified, of which 38 (29%) were community-onset. The community-onset infection group were less likely to have a recent history of antibiotic use (66% vs. 98%; p<0.001) or proton pump inhibitor use (38% vs. 69%; p=0.03) than the hospital-acquired infection group. Markers of severity and outcomes were similar in the two groups, with an overall mortality of 9%. CONCLUSIONS: Community-onset CDI accounts for a large proportion of C. difficile infections and has a similar potential for severe disease as hospital-acquired infection. Using a history of previous antibiotic use, proton pump inhibitor use, or recent hospitalization to predict cases is unreliable. We recommend that patients with diarrhoea being investigated in emergency departments and community practice are tested for Clostridium difficile infection.
Assuntos
Clostridioides difficile , Infecções por Clostridium/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Austrália , Infecções por Clostridium/diagnóstico , Infecções por Clostridium/mortalidade , Infecções Comunitárias Adquiridas/diagnóstico , Infecções Comunitárias Adquiridas/epidemiologia , Infecção Hospitalar/diagnóstico , Infecção Hospitalar/epidemiologia , Feminino , Hospitalização , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Centros de Atenção Terciária , Adulto JovemRESUMO
BACKGROUND: Enteric protozoa are associated with diarrhoeal illnesses in humans; however there are no recent studies on their epidemiology and geographical distribution in Australia. This study describes the epidemiology of enteric protozoa in the state of New South Wales and incorporates spatial analysis to describe their distribution. DESIGN AND METHODS: Laboratory and clinical records from four public hospitals in Sydney for 910 patients, who tested positive for enteric protozoa over the period January 2007 - December 2010, were identified, examined and analysed. We selected 580 cases which had residence post code data available, enabling us to examine the geographic distribution of patients, and reviewed the clinical data of 252 patients to examine possible links between protozoa, demographic and clinical features. RESULTS: Frequently detected protozoa were Blastocystis spp. (57%), Giardia intestinalis (27%) and Dientamoeba fragilis (12%). The age distribution showed that the prevalence of protozoa decreased with age up to 24 years but increasing with age from 25 years onwards. The geographic provenance of the patients indicates that the majority of cases of Blastocystis (53.1%) are clustered in and around the Sydney City Business District, while pockets of giardiasis were identified in regional/rural areas. The distribution of cases suggests higher risk of protozoan infection may exist for some communities. CONCLUSIONS: These findings provide useful information for policy makers to design and tailor interventions to target high risk communities. Follow-up investigation into the risk factors for giardiasis in regional/rural areas is needed. Significance for public healthThis research is significant since it provides the most recent epidemiological update on the common enteric protozoa affecting Australians. It reveals that enteric protozoa cause considerable disease burden in high risk city dwellers, and provides the evidence base for development of targeted interventions for their prevention and control in high risk populations. The prevalence of enteric protozoa in this metropolitan setting underscores that microorganisms do not respect borders and that a collaborative approach is needed to contain the global spread of infectious diseases. Incorporating spatial analysis is valuable in providing a compelling picture of the geographical distribution of these often neglected diseases. Local and State Public Health departments can use this information to support further inves-
RESUMO
We aimed to streamline the diagnosis of gastrointestinal disease by producing multiplexed real time polymerase chain reaction (PCR) panels employing universal sample processing for DNA and RNA containing pathogens. A total of 487 stored, previously characterised stool samples comprising bacterial, viral, protozoan and Clostridium difficile positive samples were tested using four multiplexed real time PCR panels. A further 81 pre-selected clinical samples from a teaching hospital were included to provide an independent validation of assay performance. Improved sensitivity was achieved using the protozoan panels and 16 more mixed infections were observed compared to tests with conventional methods. Using the C. difficile panels, 100% sensitivity was achieved when compared to the gold standard of toxigenic culture. In addition, hypervirulent strains including ribotype 027 could be identified directly from primary sample without the need for ribotyping methods. Bacterial and viral panels detecting Salmonella, Shigella, Campylobacter, Yersinia enterocolitica, Listeria monocytogenes, norovirus groups I and II, rotavirus A, astrovirus, sapovirus, rotavirus B, adenovirus and adenovirus 40/41 performed as well as conventional methods, whilst allowing detection in 3 hours from processing to result. Multiplex real time PCR panels with universal sample preparation allow streamlined, rapid diagnosis of gastrointestinal pathogens whilst extending the characterisation of pathogens present in stool samples from affected patients.
Assuntos
Bactérias/isolamento & purificação , Gastroenteropatias/diagnóstico , Reação em Cadeia da Polimerase Multiplex/métodos , Parasitos/isolamento & purificação , Vírus/isolamento & purificação , Animais , Bactérias/genética , Clostridioides difficile/genética , Clostridioides difficile/isolamento & purificação , Fezes/microbiologia , Fezes/parasitologia , Gastroenteropatias/microbiologia , Gastroenteropatias/parasitologia , Humanos , Parasitos/genética , Kit de Reagentes para Diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Sensibilidade e Especificidade , Fatores de Tempo , Vírus/genéticaRESUMO
Norovirus is the leading cause of acute gastroenteritis with most infections caused by GII.4 variants. To understand the evolutionary processes that contribute to the emergence of GII.4 variants, we examined the molecular epidemiology of norovirus-associated acute gastroenteritis in Australia and New Zealand from 893 outbreaks between 2009 and 2012. Throughout the study GII.4 New Orleans 2009 was predominant; however, during 2012 it was replaced by an emergent GII.4 variant, Sydney 2012. An evolutionary analysis of capsid gene sequences was performed to determine the origins and selective pressures driving the emergence of these recently circulating GII.4 variants. This revealed that both New Orleans 2009 and Sydney 2012 share a common ancestor with GII.4 Apeldoorn 2007. Furthermore, pre-epidemic ancestral variants of each virus were identified up to two years before their pandemic emergence. Adaptive changes at known blockade epitopes in the viral capsid were also identified that likely contributed to their emergence.