The molecular characterisation of shigella spp. from Papua New Guinea and Pacific Island Nations
- Authors: Malau, Elisheba
- Date: 2019
- Type: Text , Thesis , PhD
- Full Text:
- Description: In Papua New Guinea (PNG) and other low-middle income countries (LMICs) in Oceania, diarrhoea remains a leading cause of hospitalisation and death in children <5 years old; and is an important cause of illness in older children and adults. Shigella is one of four leading causes of diarrhoea globally and is likely a major cause of diarrhoea in Oceanic LMICs, particularly PNG. A survey was conducted among parents of children with diarrhoea in the PNG highlands, demonstrating limited access to, and application of, sanitation and hygiene; likely contributing to the transmission of Shigella and other pathogens in PNG. To better understand Shigella in Oceanic LMICs, multiple analyses of isolates from within PNG (n=38), far-northeastern Australia, and from travellers returning to Australia from Oceanic LMICs with shigellosis (n=34) were conducted. Antimicrobial susceptibility (by disc diffusion), detection of virulence genes by polymerase chain reaction (PCR) were conducted on all isolates, and whole genome sequencing (WGS) conducted on 63 isolates. Shigella spp. were commonly resistant to two or more classes of first-line antibiotics, with resistance more common in post-2010 relative to pre-2010 isolates. WGS was used to verify PCR detection of virulence genes, determine whether resistance could be predicted genetically, and conduct phylogenetic analysis of Shigella spp. in Oceania. WGS surpassed PCR in the detection of virulence genes, but correlated poorly with phenotypic antimicrobial resistance. Phylogenetic analysis revealed the intra- and inter-country relatedness. Three phylogenetic groups of S. flexneri co-exist in Oceanic LMICs (and far-northeastern Australia), the result of multiple incursions. Two lineages of S. sonnei were detected, one circulating in PNG and New Caledonia, and the other in various other countries including Fiji; with no geographical overlap of the two S. sonnei lineages. Incursions of Shigella into the Oceanic LMICs occur regularly, and are likely to occur again. This study provides evidence of the need for, and potential approach to, expanded surveillance of Shigella in the region.
- Description: Doctor of Philsophy
- Authors: Malau, Elisheba
- Date: 2019
- Type: Text , Thesis , PhD
- Full Text:
- Description: In Papua New Guinea (PNG) and other low-middle income countries (LMICs) in Oceania, diarrhoea remains a leading cause of hospitalisation and death in children <5 years old; and is an important cause of illness in older children and adults. Shigella is one of four leading causes of diarrhoea globally and is likely a major cause of diarrhoea in Oceanic LMICs, particularly PNG. A survey was conducted among parents of children with diarrhoea in the PNG highlands, demonstrating limited access to, and application of, sanitation and hygiene; likely contributing to the transmission of Shigella and other pathogens in PNG. To better understand Shigella in Oceanic LMICs, multiple analyses of isolates from within PNG (n=38), far-northeastern Australia, and from travellers returning to Australia from Oceanic LMICs with shigellosis (n=34) were conducted. Antimicrobial susceptibility (by disc diffusion), detection of virulence genes by polymerase chain reaction (PCR) were conducted on all isolates, and whole genome sequencing (WGS) conducted on 63 isolates. Shigella spp. were commonly resistant to two or more classes of first-line antibiotics, with resistance more common in post-2010 relative to pre-2010 isolates. WGS was used to verify PCR detection of virulence genes, determine whether resistance could be predicted genetically, and conduct phylogenetic analysis of Shigella spp. in Oceania. WGS surpassed PCR in the detection of virulence genes, but correlated poorly with phenotypic antimicrobial resistance. Phylogenetic analysis revealed the intra- and inter-country relatedness. Three phylogenetic groups of S. flexneri co-exist in Oceanic LMICs (and far-northeastern Australia), the result of multiple incursions. Two lineages of S. sonnei were detected, one circulating in PNG and New Caledonia, and the other in various other countries including Fiji; with no geographical overlap of the two S. sonnei lineages. Incursions of Shigella into the Oceanic LMICs occur regularly, and are likely to occur again. This study provides evidence of the need for, and potential approach to, expanded surveillance of Shigella in the region.
- Description: Doctor of Philsophy
Antimicrobial sensitivity trends and virulence genes in Shigella spp. from the Oceania region
- Malau, Elisheba, Ford, Rebecca, Valcanis, Mary, Jennison, Amy, Mosse, Jenny, Bean, David, Yoannes, Mition, Pomat, William, Horwood, Paul, Greenhill, Andrew
- Authors: Malau, Elisheba , Ford, Rebecca , Valcanis, Mary , Jennison, Amy , Mosse, Jenny , Bean, David , Yoannes, Mition , Pomat, William , Horwood, Paul , Greenhill, Andrew
- Date: 2018
- Type: Text , Journal article
- Relation: Infection, Genetics and Evolution Vol. 64, no. (2018), p. 52-56
- Full Text:
- Reviewed:
- Description: Shigella is a common cause of diarrhoea in Papua New Guinea (PNG) and other Oceania countries. However, little is known about the strains causing infection. Archived Shigella isolates (n = 72) were obtained from research laboratories in PNG and reference laboratories in Australia. Shigella virulence genes were detected by PCR, and antimicrobial susceptibility was determined by disk diffusion. The ipaH virulence gene was present in all 72 isolates. The prevalence of other virulence genes was variable, with ial, invE, ipaBCD, sen/ospD3 and virF present in 60% of isolates and set1A and set1B genes present in 42% of isolates. Most S. flexneri isolates contained genes encoding enterotoxin 1 and/or enterotoxin 2. Resistance to antibiotics was common, with 51/72 isolates resistant to 2–4 antimicrobials. A greater proportion of bacteria isolated since 2010 (relative to pre-2010 isolates) were resistant to commonly used antibiotics such as ampicillin, chloramphenicol, tetracycline, and trimethoprim-sulfamethoxazole; suggesting that antimicrobial resistance (AMR) in Shigella is increasing over time in the Oceania region. There is a need for improved knowledge regarding Shigella circulation in the Oceania region and further monitoring of AMR patterns. © 2018 Elsevier B.V.
- Authors: Malau, Elisheba , Ford, Rebecca , Valcanis, Mary , Jennison, Amy , Mosse, Jenny , Bean, David , Yoannes, Mition , Pomat, William , Horwood, Paul , Greenhill, Andrew
- Date: 2018
- Type: Text , Journal article
- Relation: Infection, Genetics and Evolution Vol. 64, no. (2018), p. 52-56
- Full Text:
- Reviewed:
- Description: Shigella is a common cause of diarrhoea in Papua New Guinea (PNG) and other Oceania countries. However, little is known about the strains causing infection. Archived Shigella isolates (n = 72) were obtained from research laboratories in PNG and reference laboratories in Australia. Shigella virulence genes were detected by PCR, and antimicrobial susceptibility was determined by disk diffusion. The ipaH virulence gene was present in all 72 isolates. The prevalence of other virulence genes was variable, with ial, invE, ipaBCD, sen/ospD3 and virF present in 60% of isolates and set1A and set1B genes present in 42% of isolates. Most S. flexneri isolates contained genes encoding enterotoxin 1 and/or enterotoxin 2. Resistance to antibiotics was common, with 51/72 isolates resistant to 2–4 antimicrobials. A greater proportion of bacteria isolated since 2010 (relative to pre-2010 isolates) were resistant to commonly used antibiotics such as ampicillin, chloramphenicol, tetracycline, and trimethoprim-sulfamethoxazole; suggesting that antimicrobial resistance (AMR) in Shigella is increasing over time in the Oceania region. There is a need for improved knowledge regarding Shigella circulation in the Oceania region and further monitoring of AMR patterns. © 2018 Elsevier B.V.
Antibiotic resistant Shigella is a major cause of diarrhoea in the Highlands of Papua New Guinea
- Greenhill, Andrew, Guwada, Carlton, Siba, Valentine, Michael, Audrey, Yoannes, Mution, Wawarie, Yolandah, Ford, Rebecca, Siba, Peter, Horwood, Paul
- Authors: Greenhill, Andrew , Guwada, Carlton , Siba, Valentine , Michael, Audrey , Yoannes, Mution , Wawarie, Yolandah , Ford, Rebecca , Siba, Peter , Horwood, Paul
- Date: 2014
- Type: Text , Journal article
- Relation: Journal of Infection in Developing Countries Vol. 8, no. 11 (2014), p. 1391-1397
- Full Text:
- Reviewed:
- Description: Introduction: Diarrhoea remains a major cause of illness in Papua New Guinea (PNG); however, little is known about its aetiology. As a result of the cholera outbreak that spread throughout PNG in 2009-2011, we conducted diarrhoeal surveillance in Eastern Highlands Province. Methodology: Following informed consent and a brief questionnaire, participants provided a stool sample or duplicate rectal swabs. Samples were tested for common bacterial pathogens Salmonella spp., Shigella spp., Vibrio spp., Campylobacter spp. and Yersinia enterocolitica using established culture methods. Enteric parasites were detected using microscopy. Results: A total of 216 participants were enrolled; where age was recorded, 42% were under 5 years of age, 6.7% were 5 to 17 years of age and 51.3% ≥18 years of age. One or more pathogens were detected in 68 (31.5%) participants, with Shigella (primarily S. flexneri) being the most commonly isolated (47 of 216 participants). Enteric parasites were detected in 23 of the 216 participants, occurring as a co-infection with another pathogen in 12 of 23 cases. No Vibrio cholerae was detected. Shigella isolates were commonly resistant to ampicillin, tetracycline, co-trimoxazole and chloramphenicol. Conclusions: Shigellae, specifically S. flexneri, are important pathogens in the highlands of PNG. While most studies in low-income settings focus on childhood aetiology, we have demonstrated the importance of Shigella in both children and adults. Enteric parasites remain present and presumably contribute to the burden of gastrointestinal illness. While improvements in sanitation and hygiene would help lower the burden of all aetiologies of infectious diarrhoea, additional control strategies targeting Shigella may also be warranted. © 2014 Greenhill et al.
- Authors: Greenhill, Andrew , Guwada, Carlton , Siba, Valentine , Michael, Audrey , Yoannes, Mution , Wawarie, Yolandah , Ford, Rebecca , Siba, Peter , Horwood, Paul
- Date: 2014
- Type: Text , Journal article
- Relation: Journal of Infection in Developing Countries Vol. 8, no. 11 (2014), p. 1391-1397
- Full Text:
- Reviewed:
- Description: Introduction: Diarrhoea remains a major cause of illness in Papua New Guinea (PNG); however, little is known about its aetiology. As a result of the cholera outbreak that spread throughout PNG in 2009-2011, we conducted diarrhoeal surveillance in Eastern Highlands Province. Methodology: Following informed consent and a brief questionnaire, participants provided a stool sample or duplicate rectal swabs. Samples were tested for common bacterial pathogens Salmonella spp., Shigella spp., Vibrio spp., Campylobacter spp. and Yersinia enterocolitica using established culture methods. Enteric parasites were detected using microscopy. Results: A total of 216 participants were enrolled; where age was recorded, 42% were under 5 years of age, 6.7% were 5 to 17 years of age and 51.3% ≥18 years of age. One or more pathogens were detected in 68 (31.5%) participants, with Shigella (primarily S. flexneri) being the most commonly isolated (47 of 216 participants). Enteric parasites were detected in 23 of the 216 participants, occurring as a co-infection with another pathogen in 12 of 23 cases. No Vibrio cholerae was detected. Shigella isolates were commonly resistant to ampicillin, tetracycline, co-trimoxazole and chloramphenicol. Conclusions: Shigellae, specifically S. flexneri, are important pathogens in the highlands of PNG. While most studies in low-income settings focus on childhood aetiology, we have demonstrated the importance of Shigella in both children and adults. Enteric parasites remain present and presumably contribute to the burden of gastrointestinal illness. While improvements in sanitation and hygiene would help lower the burden of all aetiologies of infectious diarrhoea, additional control strategies targeting Shigella may also be warranted. © 2014 Greenhill et al.
Detection of enteric viral and bacterial pathogens associated with paediatric diarrhoea in Goroka, Papua New Guinea
- Soli, Kevin, Maure, Tobias, Kas, Monalisa, Bande, Grace, Bebes, Sauli, Luang-Suarkia, Dagwin, Siba, Peter, Morita, Ayako, Umezaki, Masahiro, Greenhill, Andrew, Horwood, Paul
- Authors: Soli, Kevin , Maure, Tobias , Kas, Monalisa , Bande, Grace , Bebes, Sauli , Luang-Suarkia, Dagwin , Siba, Peter , Morita, Ayako , Umezaki, Masahiro , Greenhill, Andrew , Horwood, Paul
- Date: 2014
- Type: Text , Journal article
- Relation: International Journal of Infectious Diseases Vol. 27, no. (2014), p. 54-58
- Full Text:
- Reviewed:
- Description: Objectives: The aim of this study was to investigate the viral and bacterial causes of acute watery diarrhoea in hospitalized children in Papua New Guinea. Methods: A retrospective analysis was conducted on stool samples collected from 199 children (age > 5 years) admitted to the paediatric ward of Goroka General Hospital from August 2009 through November 2010. A large range of viral and bacterial enteric pathogens were targeted using real-time PCR/RT-PCR assays. Results: Young children were much more likely to be admitted with acute gastroenteritis, with 62.8% of patients aged >1 year and 88.4% aged >2 years. An enteric pathogen was detected in 69.8% (n= 138) of patients. The most commonly detected pathogens were Shigella spp (26.6%), rotavirus (25.6%), adenovirus types 40/41 (11.6%), enterotoxigenic Escherichia coli (11.1%), enteropathogenic E. coli (8.5%), norovirus G2 (6.0%), and Campylobacter spp (4.0%). Norovirus G1, sapovirus, and Salmonella spp were also detected, but below our statistical limit of detection. Vibrio cholerae and astrovirus were not detected in any patients. Mixed infections were detected in 22.1% of patients, with Shigella and rotavirus most commonly detected in co-infections with other pathogens. Conclusions: This study demonstrates that Shigella and rotavirus are the major pathogens associated with acute paediatric gastroenteritis in this setting. © 2014 The Authors.
- Authors: Soli, Kevin , Maure, Tobias , Kas, Monalisa , Bande, Grace , Bebes, Sauli , Luang-Suarkia, Dagwin , Siba, Peter , Morita, Ayako , Umezaki, Masahiro , Greenhill, Andrew , Horwood, Paul
- Date: 2014
- Type: Text , Journal article
- Relation: International Journal of Infectious Diseases Vol. 27, no. (2014), p. 54-58
- Full Text:
- Reviewed:
- Description: Objectives: The aim of this study was to investigate the viral and bacterial causes of acute watery diarrhoea in hospitalized children in Papua New Guinea. Methods: A retrospective analysis was conducted on stool samples collected from 199 children (age > 5 years) admitted to the paediatric ward of Goroka General Hospital from August 2009 through November 2010. A large range of viral and bacterial enteric pathogens were targeted using real-time PCR/RT-PCR assays. Results: Young children were much more likely to be admitted with acute gastroenteritis, with 62.8% of patients aged >1 year and 88.4% aged >2 years. An enteric pathogen was detected in 69.8% (n= 138) of patients. The most commonly detected pathogens were Shigella spp (26.6%), rotavirus (25.6%), adenovirus types 40/41 (11.6%), enterotoxigenic Escherichia coli (11.1%), enteropathogenic E. coli (8.5%), norovirus G2 (6.0%), and Campylobacter spp (4.0%). Norovirus G1, sapovirus, and Salmonella spp were also detected, but below our statistical limit of detection. Vibrio cholerae and astrovirus were not detected in any patients. Mixed infections were detected in 22.1% of patients, with Shigella and rotavirus most commonly detected in co-infections with other pathogens. Conclusions: This study demonstrates that Shigella and rotavirus are the major pathogens associated with acute paediatric gastroenteritis in this setting. © 2014 The Authors.
- Soli, Kevin, Kas, Monalisa, Maure, Tobias, Umezaki, Masahiro, Morita, Ayako, Siba, Peter, Greenhill, Andrew, Horwood, Paul
- Authors: Soli, Kevin , Kas, Monalisa , Maure, Tobias , Umezaki, Masahiro , Morita, Ayako , Siba, Peter , Greenhill, Andrew , Horwood, Paul
- Date: 2013
- Type: Text , Journal article
- Relation: Diagnostic Microbiology and Infectious Disease Vol. 77, no. 4 (2013), p. 321-323
- Full Text: false
- Reviewed:
- Description: We evaluated loop-mediated isothermal amplification end-point detection methods for Salmonella, Shigella, and Vibrio cholerae. Detection sensitivities were comparable to real-time PCR methods. The colorimetric dyes hydroxynaphthol blue and SYBR Green I showed increased sensitivity when compared to visual and automated turbidity readings. End-point colorimetric dyes promise great utility in developing settings.
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