Otitis media susceptibility and shifts in the head and neck microbiome due to SPINK5 variants.

BACKGROUND: Otitis media (OM) susceptibility has significant heritability; however, the role of rare variants in OM is mostly unknown. Our goal is to identify novel rare variants that confer OM susceptibility. METHODS: We performed exome and Sanger sequencing of >1000 DNA samples from 551 multiethnic families with OM and unrelated individuals, RNA-sequencing and microbiome sequencing and analyses of swabs from the outer ear, middle ear, nasopharynx and oral cavity. We also examined protein localisation and gene expression in infected and healthy middle ear tissues. RESULTS: A large, intermarried pedigree that includes 81 OM-affected and 53 unaffected individuals cosegregates two known rare A2ML1 variants, a common FUT2 variant and a rare, novel pathogenic variant c.1682A>G (p.Glu561Gly) within SPINK5 (LOD=4.09). Carriage of the SPINK5 missense variant resulted in increased relative abundance of Microbacteriaceae in the middle ear, along with occurrence of Microbacteriaceae in the outer ear and oral cavity but not the nasopharynx. Eight additional novel SPINK5 variants were identified in 12 families and individuals with OM. A role for SPINK5 in OM susceptibility is further supported by lower RNA counts in variant carriers, strong SPINK5 localisation in outer ear skin, faint localisation to middle ear mucosa and eardrum and increased SPINK5 expression in human cholesteatoma. CONCLUSION: SPINK5 variants confer susceptibility to non-syndromic OM. These variants potentially contribute to middle ear pathology through breakdown of mucosal and epithelial barriers, immunodeficiency such as poor vaccination response, alteration of head and neck microbiota and facilitation of entry of opportunistic pathogens into the middle ear.

Examining the Evidence for an Adult Healthy Middle Ear Microbiome.

Otitis media (OM) is a cluster of diseases of the middle ear that commonly result from bacterial infection. OM subtypes in which the tympanic membrane is intact (acute otitis media and otitis media with effusion) are presumed to result from pathogen translocation through the eustachian tube. Recent molecular-based studies have suggested that a diverse middle ear microbiome exists in the absence of disease. These have been largely unsupported by culture and feature species that commonly contaminate low-biomass sequencing data. Combining culture-based and molecular techniques, we undertook a detailed investigation of the evidence for bacterial colonization of the healthy middle ear. Middle ear (ME), nasopharynx (NP), and external ear canal (EC) swabs were collected from a total of 25 adult patients undergoing cochlear implant, stapedotomy, or translabyrinthine vestibular schwannoma resection. Diagnostic culture, microscopy, quantitative PCR, and 16S rRNA gene amplicon sequencing were used to assess sample bacterial content. EC and NP microbiota were consistent with previous reports. In contrast, bacterial levels in ME samples were not significantly above those in unused control swabs. Commonly detected taxa were among recognized sequencing contaminants (Methylobacterium, Pseudomonas, and Acinetobacter). Linear regression of dominant ME taxa confirmed a negative relationship between relative abundance and bacterial load, consistent with contamination. No bacteria were detected by microscopy or diagnostic culture in any middle ear sample. Our findings cast substantial doubt on previous reports identifying a healthy middle ear microbiome using 16S amplicon sequencing.IMPORTANCE Recent molecular-based studies have suggested that a diverse middle ear microbiome in adults and children can exist in the absence of disease. These studies have been largely unsupported by culture and feature species that commonly contaminate low-biomass sequencing data. While 16S rRNA gene amplicon sequencing has proven to be a highly informative technique in many clinical contexts, it is susceptible to spurious signal arising from sequencing reagent contaminants where sample biomass is low. Combining culture-based and molecular techniques, we undertook a detailed investigation of the evidence for bacterial colonization of the healthy middle ear. In finding no evidence of viable bacterial cells in middle ear samples, our study further underlines the importance of careful consideration of amplicon sequence data derived from very-low-biomass contexts and the value of analytical approaches that combine culture and molecular techniques.

Bartonella henselae in a dog with ear tip vasculitis.

BACKGROUND: Bartonella henselae, a Gram-negative, zoonotic, alpha-proteobacteria has been previously implicated in association with cutaneous vasoproliferative lesions (bacillary angiomatosis), nodular panniculitis and multifocal erythema (erythema multiforme) in dogs. OBJECTIVE: Describe clinical, microbiological and histological lesions in a dog with ear margin vasculitis and B. henselae infection. ANIMALS: A 12-month-old, specific pathogen-free intact female beagle dog maintained in a vector-free laboratory animal resource facility. METHODS AND MATERIALS: Bartonella and Rickettsia serological evaluation, Bartonella and Rickettsia PCR, Bartonella alpha-proteobacteria growth medium (BAPGM) enrichment blood culture/PCR, histopathological investigation and confocal immunohistochemical evaluation. RESULTS: Serological investigation (seroreversion) and PCR testing of aural tissue biopsies failed to support Rickettsia rickettsii as a cause of the aural vasculitis; however, B. henselae, genotype San Antonio 2 DNA was amplified and sequenced from both ear tip margins and from normal-appearing abdominal skin. Seroconversion to B. henselae was documented retrospectively by IFA testing. Bartonella henselae organisms were visualized by confocal immunostaining within all three biopsies. Histopathology revealed small vessel necrotizing vasculitis and dermal necrosis. Bartonella henselae seroreversion and complete resolution of skin lesions occurred in conjunction with administration of oral doxycycline and enrofloxacin for six weeks. CONCLUSIONS AND CLINICAL IMPORTANCE: Bartonella henselae is an emerging zoonotic pathogen that has been associated with leucocytoclastic vasculitis in humans and may have had a contributing or causative role in the development of the cutaneous aural margin vasculitis in this beagle.

Scedosporium apiospermum: a rare cause of malignant otitis externa.

A 79-year-old man, with a history of well-controlled diabetes mellitus, presented with left-sided otalgia. With an initial diagnosis of simple otitis externa, he was discharged on topical drops. He represented 2 months later with worsening otalgia and discharge. A diagnosis of malignant otitis externa was made based on clinical and radiological findings. Intravenous Tazocin and Gentamicin were given based on previous bacterial culture from ear swabs. The patient failed to improve and developed left-sided facial nerve palsy. His condition stabilised following a change in antimicrobial therapy and his management continued in the community on intravenous Meropenem with twice weekly aural toilet. Repeated nuclear medicine imaging failed to demonstrate resolution. A bony sequestration was removed from the external auditory canal in the outpatient clinic, which following extended culture grew Scedosporium apiospermum; his management was subsequently changed to oral Voriconazole. This led to rapid clinical improvement and disease resolution over a 6 -week period.

[Cutaneous tuberculosis of the ear due to Mycobacterium bovis]. / Tuberculose cutanée de l'oreille à Mycobacterium bovis.

INTRODUCTION: Isolated cutaneous tuberculosis is uncommon, accounting for only 0.14 to 5% of Mycobacterium tuberculosis infections. We report a rare case of ear cutaneous tuberculosis due to Mycobacterium bovis in an immunocompetent woman. CASE REPORT: A 59-year-old woman presented an erythematous and scaly lesion of the ear present for two years. The histological findings were compatible with a diagnosis of sarcoidosis, with non-necrotic granuloma. After failure of dermal corticosteroid therapy, a further biopsy identified M. bovis; the patient was cured following anti-tubercular treatment. DISCUSSION: Ear lesions are predominantly associated with tumors, fungal infections, chondritis, lupus and sarcoidosis. The ear, like the face in general, is a classic localization of lupus vulgaris, a chronic form of confined tuberculosis infection with progressive evolution. The paucibacillary nature of these lesions is the reason why their diagnosis is based in some cases on clinical, histological and immunological findings without bacteriological evidence. However, given the potential therapeutic implications, it is important to push the microbiological analysis as far as possible. In our case, culture and identification provided evidence of M. bovis infection, enabling suitable and effective therapy to be given.

Comparative study of aural microflora in healthy cats, allergic cats and cats with systemic disease.

Twenty healthy cats (group 1) with clinically normal ears, 15 cats with systemic disease (group 2) and 15 allergic cats (group 3) were included in a prospective study. The experimental unit was the ear. A clinical score was established for each ear canal after otoscopic examination. Microbial population was assessed on cytological examination of smears performed with the cotton-tipped applicator smear technique. Fungal population was significantly more prominent in allergic cats (P <0.001) and in diseased cats compared with healthy cats (P <0.02). Bacterial population was significantly higher in allergic cats than in healthy cats (P <0.001) and cats suffering from systemic disease (P <0.001). Bacterial overgrowth was also higher in cats with systemic disease than healthy cats. In cats from group 2, only fungal overgrowth was associated with otitis severity. In group 3, only bacterial overgrowth was associated with otitis severity.

Lepromatous leprosy with an uncommon presentation: a case report.

28 yr old male presented with asymptomatic nodules and few well to ill defined papules on ears, asymmetrical nerve enlargement and evanescent tender nodules on the extremities without any infiltration of the skin and madarosis. Slit skin smear done from normal skin was BI 6+. Skin biopsy showed features of lepromatous leprosy.

Epicutaneous model of community-acquired Staphylococcus aureus skin infections.

Staphylococcus aureus is one of the most common etiological agents of community-acquired skin and soft tissue infection (SSTI). Although the majority of S. aureus community-acquired SSTIs are uncomplicated and self-clearing in nature, some percentage of these cases progress into life-threatening invasive infections. Current animal models of S. aureus SSTI suffer from two drawbacks: these models are a better representation of hospital-acquired SSTI than community-acquired SSTI, and they involve methods that are difficult to replicate. For these reasons, we sought to develop a murine model of community-acquired methicillin-resistant S. aureus SSTI (CA-MRSA SSTI) that can be consistently reproduced with a high degree of precision. We utilized this model to begin to characterize the host immune response to this type of infection. We infected mice via epicutaneous challenge of the skin on the outer ear pinna using Morrow-Brown allergy test needles coated in S. aureus USA300. When mice were challenged in this model, they developed small, purulent, self-clearing lesions with predictable areas of inflammation that mimicked a human infection. CFU in the ear pinna peaked at day 7 before dropping by day 14. The T(h)1 and T(h)17 cytokines gamma interferon (IFN-γ), interleukin-12 (IL-12) p70, tumor necrosis factor alpha (TNF-α), IL-17A, IL-6, and IL-21 were all significantly increased in the draining lymph node of infected mice, and there was neutrophil recruitment to the infection site. In vivo neutrophil depletion demonstrated that neutrophils play a protective role in preventing bacterial dissemination and fatal invasive infection.

[Reliability and agreement of two smear reading scales for classification and monitoring of multidrug therapy in leprosy patients]. / Confiabilidad y concordancia de dos escalas de lectura de baciloscopias para clasificación y seguimiento del tratamiento con múltiples medicamentos de los pacientes con lepra.

INTRODUCTION: After the clinical diagnosis of leprosy, classification methods are necessary to define a treatment and prognosis of patients consistent with bacterial load. Bacteria are detected in skin smear, and bacterial load typically is established by the internationally used Ridley's logarithmic scale, However, in Colombia an alternative semiquantitative scale is used. OBJECTIVE: The interobserver reproducibility was established for the Ridley and Colombia scales, and the level of correlation-matching was identified between the bacillary indices obtained in order to assess the degree of interchangeability. MATERIALS AND METHODS: Standardization was attained by a reading of the smears by 2 readers with subsequent, blinded evaluation of inter-observer agreement. Each reader quantified the bacterial load of for each sample (n=325) using the Colombian and the Ridley scales. The degree of interobserver agreement was assessed with weighted kappa coefficient. The level of correlation and agreement between the measurements of the bacillary index was established with coefficient of Lin. RESULTS: The interobserver weighted kappa coefficient was 0.83 for the Colombia scale and 0.85 for the Ridley scale. The Lin coefficient was 0.96 for the correlation-matching of bacillary indexes. CONCLUSIONS: Interobserver agreement obtained for both scales was excellent as the correlation-matching bacillary indices determined with both methods. With the cut-off points yielded a good level of agreement, ensuring interchangeability between the scales defining the high or low bacterial load.

An analysis of the bacterial flora found in the external auditory canals of microtia patients: results and clinical applications.

During the process of auricular reconstruction in cases of microtia patients with external auditory canals (EAC), bacterial contamination from the EAC can cause cartilage infection. In this article, we retrospectively analyzed the data on bacterial flora present in the EAC of these patients.Preoperatively, in 91 microtia patients, culture samples were obtained, and isolates were tested for in vitro susceptibility to antibiotics. One hundred volunteers with no ear abnormalities were also evaluated as part of the control group.Seventy-nine specimens in 91 microtia patients showed growth of bacteria (86.8%): a total of 97 organisms were isolated. Dominating bacteria were of the staphylococci species (85.6%). Fourteen Staphylococcus isolates were resistant to methicillin. The percentage of microtia patients showing a presence of methicillin-resistant staphylococci isolates (15%) was significantly higher than the percentage in the "normal" volunteer group showing a presence of methicillin-resistant staphylococci isolates (2%; P = 0.0009).To decrease the complications that occurred due to cartilage infection during auricular reconstruction in the cases of microtia with EAC, we suggest that bacterial floras of the EAC be routinely examined and the patients be treated with appropriate antibiotics preoperatively.

Canadian case report of erythema nodosum leprosum successfully treated with prednisone and thalidomide.

BACKGROUND: Erythema nodosum leprosum (ENL) is a disease rarely encountered in Canada. It is characterized by multiple remissions and recurrences, often requires long-term treatment, and can result in debilitating sequelae. OBJECTIVE: To promote rapid recognition and adequate therapy for ENL. METHODS: Case report of a 39-year-old man diagnosed with an ENL. The clinical and histopathologic features, treatment provided, and response to treatment are detailed in this article. RESULTS: ENL presented itself as painful cutaneous lesions on the face and limbs, bilateral paresthesia of the fourth and fifth fingers, and systemic symptoms. Prednisone 40 mg daily for a week and then 60 mg daily for another week reduced the lesions by 80% and the pain by 50%. Although prednisone 60 mg daily was continued for one more week and then stopped, thalidomide was started at a dose of 300 mg daily for 4 weeks and then reduced gradually, which led to complete resolution. CONCLUSION: At the 7(1/2)-month follow-up, the patient remained completely asymptomatic.

[Pigs as an infection source for methicillin resistant Staphylococcus aureus infections in humans]. / Svin som smittekilde til infektioner med methicillinresistente Staphylococcus aureus hos mennesker.

Recent Dutch studies indicate that methicillin-resistant Staphylococcus aureus (MRSA) sequence type 398 is widely distributed in pigs and may give rise to infection in humans. In this study we present the first two Danish cases of MRSA infection, which in all probability were acquired from occupational contact with pigs. One infection presented as a severe surgical wound infection, following knee surgery, the other as a superficial ear lobe infection. Both MRSA strains were multiresistant, sequence type 398, Spa-type t034, and Panton-Valentine leukocidin-encoding gene negative.

A review of common dermatologic disorders of the external ear.

Skin disorders of the external ear are common. Although audiologists will not necessarily treat these conditions, it is important for them to be aware of these disorders and refer patients to a specialist in some instances. This report summarizes eight of the most commonly encountered skin conditions with an emphasis on recognition and appropriate referral. The cutaneous disorders of the external ear discussed in the article are divided into benign, premalignant, and malignant groups.

Malignant otitis externa.

Malignant otitis externa is an invasive, potentially life-threatening infection of the external ear and skull base that requires urgent diagnosis and treatment. It affects immunocompromised individuals, particularly those who have diabetes. The most common causative agent remains Pseudomonas aeruginosa. Definitive diagnosis is frequently elusive, requiring a high index of suspicion, various laboratory and imaging modalities, and histologic exclusion of malignancy. Long-term oral antipseudomonal agents have proven effective; however, pseudomonal antibiotic resistance patterns have emerged and therefore other bacterial and fungal causative agents must be considered. Adjunctive therapies, such as aggressive debridement and hyperbaric oxygen therapy, are reserved for extensive or unresponsive cases.

Microbiota indígena de seres humanos / Indigenous microbiota of human beings

As superfícies do corpo humano são colonizadas por uma comunidade de organismos, principalmente bactérias, que constitui a microbiota indígena. A composição desta microbiota se altera ao longo da vida e é influenciada por diversos fatores, tais como dieta e status imunológico do hospedeiro. A microbiota pode agir de maneira benéfica ou, em algumas situações, pode ser prejudicial para o indivíduo. Podem ser distinguidas a microbiota residente, constituída por organismos específicos, encontrados, freqüentemente, em determinadas áreas e a microbiota transitória, que consiste de microrganismos provenientes do ambiente, que habitam a pele e as superfícies mucosas por horas ou poucas semanas. Trato gastrointestinal, vagina, cavidade oral e pele possuem a microbiota mais rica e diversificada do corpo humano. O conhecimento da constituição da microbiota indígena é extremamente relevante para os médicos, principalmente para orientar a interpretação de resultados de exames microbiológicos e a escolha da terapia antimicrobiana empírica mais adequada. Deve-se salientar que a microbiota geralmente é benéfica. Por esse motivo, é fundamental que se tenha consciência dos riscos do rompimento da homeostasia entre microbiota e hospedeiro.

Body surfaces are colonized by a community of organisms that are recognized as indigenous microbiota, that is mainly constituted by bacteria. Its constitution changes with time and is influenced by several conditions such as diet and the immune status of the individual, among others. There are now evidences that the microbiota could be beneficial or, in some instances, dangerous to human health. It could be classified as resident, composed by fixed organisms, frequently found in certain areas, or as transitory, consisting of organisms from the environment that inhabits skin and mucosa for hours to few weeks. The gastrointestinal tract, vagina, oral cavity and skin show the richest and most diverse microbiota of the human body. The knowledge of the constitution of the indigenous microbiota is extremely important for clinicians, mainly because it can help them to interpret results of microbiological tests and to choose appropriate empirical therapy. It should be pointed out that microbiota is, in general, harmless and beneficial; for these reason, physicians must keep in mind that the disruption of the homeostasis between microbiota and host should be avoided.

Malignant external otitis: treatment with prolonged usage of antibiotics and Burow's solution.

Malignant external otitis is a severe infection of the external auditory canal, generally caused by Pseudomonas aeruginosa in elderly diabetics patients. We describe a case of malignant external otitis in a 63-year-old man with severe otalgia, purulent otorrhea and polypoid granulation of the external auditory canal. Local debridement, insulin treatment and 6-week intravenous antibiotic therapy with carbapemens were very effective and the granulation tissue disappeared completely. However, the patient readmitted for recurrent disease 3 weeks later, despite the oral and ear drop administration with new quinolone. No recurrence has been noted after 4-week additional treatment with intravenous carbapenems followed by 3-week treatment with Burow's solution as ear drops.

Isolation of Malassezia species from healthy cats and cats with otitis.

Lipid-dependent Malassezia species have recently been cultured from veterinary specimens. The identification of Malassezia species isolates from animals is important to clarify the epidemiology of these lipophilic yeasts. Malassezia species were cultured from the external ear canals of 63 out of 99 cats with otitis and 12 of 52 (23%) healthy control cats. The rate of isolation in affected animals versus controls was highly significant (P<0.01). Malassezia pachydermatis was isolated as a pure culture in 33 (45.2%) cats, associated with Malassezia globosa and Malassezia furfur in 20 (50%) and 17 (42.5%) animals, respectively. Three different species were isolated simultaneously in three cats (two cats with M pachydermatis, M globosa and M furfur, one subject with M pachydermatis, M furfur and Malassezia sympodialis). M globosa was isolated as the sole species in two animals. The present work confirms the presence of some lipid-dependent species of Malassezia in both healthy and otitic cats.

[Auricular perichondritis in adults. An evaluation-study of 12 hospitalized patients]. / Pericondritis del pabellón auricular en adultos. Estudio-evaluación de 12 pacientes ingresados.

Perichondritis of the auricle in adults can be secondary to traumatic agents, external otitis or surgery being Pseudomona aeruginosa the bacteria more often isolated. We have performed a retrospective study of 12 hospitalized patients with that diagnosis and these following variables have been evaluated: Age, sex, personal antecedents, clinical symptoms, affected auricle, complementary explorations, treatment, evolution and stay.