Measuring nasal bacterial load and its association with otitis media

Smith‐Vaughan, Heidi; Byun, Roy; Nadkarni, Mangala A.; Jacques, Nicholas A.; Hunter, Neil; Halpin, Stephen; Morris, Peter S.; Leach, Amanda J.

doi:10.1186/1472-6815-6-10

Cited by 129 publications

(146 citation statements)

References 26 publications

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“…The data shown in Table 2 and Figures 4 and 5 were obtained using an earlier version of the lytA qPCR primers and probe 17 . The current protocol describes the updated qPCR assay.…”

Section: Representative Resultsmentioning

confidence: 99%

“…Collect data and quantify pneumococcal DNA using the standard curve. Bacterial loads are calculated as previously described 17 , estimating that 1 pg of genomic DNA is equivalent to 447.4 cells, and reported as CFU/ml (assuming one genome per CFU, and one copy of lytA per genome; see Figure 4). For a successful assay, the R 2 value should be ≥0.98 and all positive results within the range of the standard curve (cT values less than that obtained for the 0.00001 ng/μl standard curve point are considered background/lytA negative).…”

Section: Viable Count Methodsmentioning

confidence: 99%

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Investigating the Effects of Probiotics on Pneumococcal Colonization Using an <em>In Vitro</em> Adherence Assay

Dunne

Toh

John

et al. 2014

JoVE

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Adherence of Streptococcus pneumoniae (the pneumococcus) to the epithelial lining of the nasopharynx can result in colonization and is considered a prerequisite for pneumococcal infections such as pneumonia and otitis media. In vitro adherence assays can be used to study the attachment of pneumococci to epithelial cell monolayers and to investigate potential interventions, such as the use of probiotics, to inhibit pneumococcal colonization. The protocol described here is used to investigate the effects of the probiotic Streptococcus salivarius on the adherence of pneumococci to the human epithelial cell line CCL-23 (sometimes referred to as HEp-2 cells). The assay involves three main steps: 1) preparation of epithelial and bacterial cells, 2) addition of bacteria to epithelial cell monolayers, and 3) detection of adherent pneumococci by viable counts (serial dilution and plating) or quantitative real-time PCR (qPCR). This technique is relatively straightforward and does not require specialized equipment other than a tissue culture setup. The assay can be used to test other probiotic species and/or potential inhibitors of pneumococcal colonization and can be easily modified to address other scientific questions regarding pneumococcal adherence and invasion. Video LinkThe video component of this article can be found at

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“…The data shown in Table 2 and Figures 4 and 5 were obtained using an earlier version of the lytA qPCR primers and probe 17 . The current protocol describes the updated qPCR assay.…”

Section: Representative Resultsmentioning

confidence: 99%

Section: Viable Count Methodsmentioning

confidence: 99%

Investigating the Effects of Probiotics on Pneumococcal Colonization Using an <em>In Vitro</em> Adherence Assay

Dunne

Toh

John

et al. 2014

JoVE

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“…Smith and Vaughan et al suggested that nasal bacterial load should be reduced in order to control the incidence of suppurative otitis media. 30 Albu et al found that three factors were significant in predicting success rate, i.e. healthy opposite ear, a long dry period preceding the operation, and non-smoker status.…”

Section: Discussionmentioning

confidence: 99%

Factors Influencing Surgical Outcome of Myringoplasty in Tubotympanic Type of Chronic Otitis Media

Das¹,

Das²

2017

jemds

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BACKGROUNDChronic otitis media is an inflammatory process of the mucoperiosteal lining of the middle ear space and mastoid. Infection of the middle ear has been a problem encountered in the human race, and is as old as humanity itself. Chronic suppurative otitis media (CSOM) is one of the most common causes of ear infection in developing countries particularly in South-east Asian countries. Chronic middle ear disease is a major problem in India especially in the rural areas. It is estimated that 6% of Indian population suffers from chronic ear disease. The term "Tubotympanic disease" is sometimes used to describe COM with a central perforation. The two principal aims of management of tubotympanic chronic otitis media are the eradication of infection and improvement of hearing by the closure of the tympanic membrane perforation which can be achieved either conservatively or by surgery. The surgical treatment of CSOM is still controversial. Some surgeons advice only repair of central perforation of tympanic membrane in non-cholesteatomatous tubotympanic chronic otitis media using graft i.e. myringoplasty to achieve a dry ear with improved hearing. This study aims to determine the factors responsible for successful surgical outcome of myringoplasty. MATERIALS AND METHODSThis hospital-based prospective study was done in the Dept. of ENT, Assam Medical College and Hospital, Dibrugarh from 1 st January 2015 to 31 st December 2015 in diagnosed cases of tubotympanic variant of CSOM. Thirty patients were included in this study after fulfilling inclusion and exclusion criteria. The surgical outcomes were assessed in terms of hearing gain, graft uptake and dryness of ear. RESULTSThirty patients of tubotympanic type of CSOM reviewed in the study with mean age group of 35.16 + 12.75 years underwent myringoplasty. Successful graft uptake post-operatively at 3 rd followup (12 th week) was seen in 83.33%. Dry ear was achieved at the end of 12 th week in 86.67% and gain in hearing at 3 rd post-operative review was 9.90 + 1.81 dB. 5 cases had graft failure, 4 cases failed to achieve dry ear. Extremity of ages, lower socio-economic status, postoperative URTI and ET dysfunction, disease in contralateral ear were associated factors in determining graft uptake and dry ear. CONCLUSIONMyringoplasty has been established as a very effective surgical management of tubotympanic variant of CSOM. Success of myringoplasty can be measured in terms of gain in hearing, graft uptake and dry ear. Our study acknowledges that successful outcome of surgery is determined by a range of factors like lower socio-economic status particularly in rural population with unhygienic living conditions, presence of nidus of infection in the upper airway along with eustachian tube dysfunction, otitis externa, disease in contralateral ear may be considered as some of the possible factors responsible for influencing the outcomes of surgery in tubotympanic type of chronic otitis media. But we emphasised that more cases or study population for longer duration...

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“…However, suppurative OM can arrest pneumatisation of the mastoid 20 bone, and archaeological evidence in ancient fossilized skulls of both the native American (Gregg & Steele 1982) and the Greenland Inuit (Homoe et al 1996a) does suggest an increase in OM subsequent to European colonisation. Genetic variation in the nasopharyngeal flora by ethnic groups has not been studied, either in contemporary or bioarchaeological remains, but the nasopharynx of Australian Aborigine children is known to be colonized by potential otopathogens at an earlier age (Leach et al 1994) and in higher numbers (Watson et al 2006), and this is predictive of subsequent middle ear suppuration (Smith-Vaughan et al 2006). This supports the idea that Aborigine populations are colonised by variants of nasopharyngeal commensals with which they have not co-evolved, and to which they may not have the genetic repertoire to mount a directed and specific immune response.…”

Section: Ethnic Variations In Otitis Media Incidencementioning

confidence: 99%

Evolution and Otitis Media: A Review, and a Model to Explain High Prevalence in Indigenous Populations

Bhutta¹

2015

Human Biology

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Inflammation of the middle ear (otitis media) comprises a group of disorders that are highly prevalent in childhood, and indeed are amongst the most common disorders of childhood. Otitis media is also heritable, and has effects on fecundity. This means that otitis media is subject to evolution, yet the evolutionary selection forces that may determine susceptibility to otitis media have never been adequately explored.Here I undertake a critical analysis of evolutionary forces that may determine susceptibility to middle ear inflammation. These forces include those determining function of the middle ear, those affecting host immunity, and those affecting colonization by, and pathogenicity of bacteria. I review existing mathematical evolutionary models of host-pathogen interaction and co-evolution, and apply these to develop a better understanding of the complex evolutionary landscape of middle ear infection and inflammation in humans. This includes an understanding of factors Pre--print version. Visit http://digitalcommons.wayne.edu/humbiol/ after 1 October 2015 to acquire the final version.2 determining the transition between stable evolutionary strategies for host and bacterial pathogens. This understanding will be later applied to analysis of otitis media in indigenous populations.In the second part of this article, I apply the approach of population genetics to devise a new theory for the high prevalence of otitis media in certain indigenous populations: the Australian Aborigine, the Native American, the Inuit, and the Maori.I suggest that high prevalence in such groups may have occurred as a result of colonization of these previously isolated populations by European immigrants in the 15 th and 16 th Centuries. This exposed them to new strains of bacteria to which their immune system had not evolved immunity, perturbing a previously stable hostpathogen co-evolutionary state.

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Measuring nasal bacterial load and its association with otitis media

Cited by 129 publications

References 26 publications

Investigating the Effects of Probiotics on Pneumococcal Colonization Using an <em>In Vitro</em> Adherence Assay

Investigating the Effects of Probiotics on Pneumococcal Colonization Using an <em>In Vitro</em> Adherence Assay

Factors Influencing Surgical Outcome of Myringoplasty in Tubotympanic Type of Chronic Otitis Media

Evolution and Otitis Media: A Review, and a Model to Explain High Prevalence in Indigenous Populations

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