Krister Tano

Inhibition of OM pathogens by alpha-hemolytic streptococci from healthy children, children with SOM and children with rAOM

The present study was undertaken to elucidate the inhibitory activity of the normal nasopharyngeal flora against the three most common otitis media (OM) pathogens in healthy children, children with secretory otitis media (SOM) and children with recurrent otitis media (rAOM). Isolates of alpha-hemolytic streptococci (AHS) and OM pathogens were recovered from the tubal orifice in each child. The samples were taken from 20 healthy children under general anesthesia, from 19 children with SOM and 20 children with rAOM. The method used to test the bacterial interference in vitro was a modified agar overlay method. The AHS sampled from the tubal orifice of the healthy children were able to inhibit 92% of the S. peumoniae isolates, 74% of the non-typable H. influenzae isolates and 89% of the M. catarrhalis isolates. The corresponding figures for children with SOM and children with rAOM were: 73% of the S. pneumoniae isolates, 58 and 54% of the non-typable H. influenzae isolates and 86 and 89% of the M. catarrhalis isolates. The AHS from children with SOM and children with rAOM were significantly less capable of inhibiting the S. peumoniae and the H. influenzae isolates (P<0.001). There was no significant difference between the three groups of children regarding inhibitory activity against M. catarrhalis. The results suggest that the inhibitory activity of the normal bacterial flora at the tubal orifice against pneumococci and H. influenzae may be reduced in children with SOM and rAOM.

A nasal spray with alpha-haemolytic streptococci as long term prophylaxis against recurrent otitis media

Previous studies have shown that children with recurrent acute otitis media (rAOM) have significantly lower quantities of alpha-haemolytic streptococci (AHS) in the nasopharynx than healthy children. Furthermore children with otitis media have AHS with lower inhibitory activity in vitro on Streptococcus pneumoniae and non-typable Haemophilus influenzae compared with healthy children. A randomised, placebo controlled and double blind clinical study among children with rAOM was designed to determine whether or not a nasal spray, containing AHS with very good inhibitory activity on the three most common OM pathogens, could be an alternative to tympanostomy tube insertion. Forty three children under 4 years of age were included in the study. The children sprayed once daily for 4 months and were monitored for 6 months. Sixteen children in the active group and 20 children in the placebo group were evaluated. The result showed no significant differences regarding the number of episodes of AOM, with seven recurrences in the active group and eight in the placebo group. No significant changes of the nasopharyngeal flora could be detected during the study period regarding the OM pathogens. Nasal spray according to the performed schedule is not yet an alternative to tympanostomy tubes in children with rAOM. The possibility of increasing the efficacy of this ecological treatment, by using pre-treatment antibiotics, more adhesive bacteria and alternative treatment schedules is discussed.

In vitro inhibition of S. pneumoniae, nontypable H. influenzae and M. catharralis by alpha-hemolytic streptococci from healthy children

The present study aimed to investigate the inhibitory activity of the normal epipharyngeal flora against the three most common acute otitis media (AOM) pathogens in healthy children, and to study if the inhibitory activity differs between alpha-hemolytic streptococci (AHS) sampled from the tubal orifice and from those sampled from the adenoid. A total number of ten isolates of AHS were collected from the tubal orifice and the adenoid, respectively, in ten children undergoing adenoidectomy or tonsillectomy. None of the children had a history of otitis media, neither secretory otitis media (SOM) nor AOM. The method used to test the bacterial interference in vitro was a modified agar overlay method. The results showed that the AHS from nasopharynx were able to inhibit the majority of the S. pneumoniae, nontypable Haemophilus influenzae and Moraxella catharralis isolates tested. The AHS isolates from the tubal orifice inhibited growth of 93% of S. pneumoniae, 79% of H. influenzae and 84% of M. catharralis isolates. The corresponding figures among isolates from the adenoid were 76, 48 and 62%. This difference in the inhibitory capacity between the AHS isolates collected from the adenoid, compared with the AHS collected from the tubal orifice, is statistically significant (P<0.01) and implies that it is important to know the exact sampling locality before conclusions are made concerning the significance of bacterial interference in the upper airways.

Alloiococcus otitidis-otitis media pathogen or normal bacterial flora?

During the last decade a new potential otitis media pathogen, Alloiococcus otitidis, has been studied. It is still not clear whether this bacterium really is a pathogen, although it has been found in a high percentage of middle ear effusions in children. The present study aimed to investigate the presence of A. otitidis in the nasopharynx and outer ear canals, and to develop a culture method that would make it possible to isolate A. otitidis from these locations. Nasopharyngeal samples (n=129) from children below 6 years were investigated by conventional culture on blood agar plates with 6% saline and rabbit antisera against A. otitidis, and by a PCR method. In the same way, we investigated 10 samples from vestibulum nasi of healthy persons, 68 samples from outer ear canals of patients with acute or chronic ear problems, and 24 samples from outer ear canals of healthy persons. In a rat model of acute otitis media, we instilled living A. otitidis into rat middle ears through the tympanic bulla and evaluated the outcome clinically by otomicroscopy at days 3, 6 and 14. Of the 129 nasopharyngeal cultures, 9 were positive for A. otitidis by PCR, but none by the culture method. Of the 68 samples from patients with running ears, 4 were positive for A. otitidis by PCR, but none by the culture method. Of the 24 healthy ear canals, 7 were positive for A. otitidis by PCR and 3 of them also by the culture method. No A. otitidis could be found from the vestibulum nasi. The rat experiment showed that the reactions in the middle ears were mild; we could not provoke a purulent acute otitis media in any of the rats. There was a 7% prevalence of A. otitidis in children below 6 years. The highest prevalence (29%) was found in outer ear canals of healthy persons, which strongly suggests that A. otitidis is part of the normal bacterial flora of the outer ear canal. The doubtful pathogenicity is also confirmed by the fact that—in the rat model—A. otitidis elicited only a mild response in the middle ear. It was possible to isolate A. otitidis using a blood agar plate with 6% saline.

A daily nasal spray with saline prevents symptoms of rhinitis

Objective —To ascertain whether a daily nasal spray with physiological saline could prevent symptoms of common cold in a population of otherwise healthy adults. Material and Methods —This was study involving 10 weeks of daily use of a nasal saline spray and 10 weeks of only recording symptoms. Young adults eligible for military service at an army barrack in Boden, Sweden were invited to participate in the study and 108 healthy conscripts aged ≈20 years agreed to do so. Data were recorded by the participants in a diary at home. In the diary the participants noted symptoms such as rhinitis, blocked nose, cough, fever and sore throat (pharyngeal pain). They also recorded inability to perform their duties due to the symptoms, and any medication or antibiotics necessitated by upper respiratory tract infection. Results —A total of 69 subjects completed the 20-week diary period. For 60 of them, compliance during the spray period exceeded 60% and their data were used in the statistical calculations. During the spray period the number of days with nasal secretion and/or blocked nose (mean 6.4 days) was significantly (p=0.027) lower than that during the observation period (mean 11 days). Furthermore, the participants had a mean of 0.7 episodes of upper respiratory tract infection during the spray period, compared with 1.0 episodes during the observation period (p=0.05). Conclusion —A daily nasal spray with saline can prevent nasal symptoms of common cold in a population of otherwise healthy adults.

The Gelfoam® plug: an alternative treatment for small eardrum perforations.

To examine if a Gelfoam® plug in combination with surgical removal of the perforation edges could be an alternative to the widely accepted fat plug treatment for smaller ear drum perforations.

Bacterial Interference Between Pathogens in Otitis Media and Alpha-Haemolytic Streptococci Analysed in an In Vitro Model

Bacterial interference studied by means of agar methods has shown a decreased number of inhibitory alpha-haemolytic Streptococci among otitis-prone children. Additional information was gained regarding the interplay between alphahaemolytic Streptococci (AHS) and otitis media (OM) pathogens by comparing the bacterial interference in broth with the interference activity studied using agar overlay methods. We found that non-typeable Haemophilus influenzae (NTHI) and Moraxella catarrhalis are readily inhibited by AHS in broth. Streptococcus pneumoniae was more bacteriostatically inhibited. If two OM pathogens were inoculated simultaneously, an isolate of AHS with poor inhibitory activity was not able to inhibit the growth, in contrast to an isolate of AHS with good inhibitory activity. The initial amount of AHS inoculated with M. catarrhalis seemed to play a decisive role with respect to the inhibitory activity. M. catarrhalis developed reduced susceptibility against AHS both in vivo and in vitro . In vivo studies showed that children with secretory otitis media had fewer isolates of AHS in their nasopharynx with the ability to inhibit all the test pathogens than healthy children ( p < 0.001). Although the factor(s) responsible for the inhibitory activity have thus far not been defined, we could exclude low pH and nutrition depletion as the inhibitory mechanism of AHS with good inhibitory activity.

Is hydrogen peroxide responsible for the inhibitory activity of alpha-haemolytic streptococci sampled from the nasopharynx?

Objective—The inhibitory effect of alpha-haemolytic Streptococci (AHS) in vitro on the three commonest otitis media pathogens, Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis, was previously investigated. The aim of this study was to determine the mechanism of this inhibitory activity. Material and Methods—When fractions of AHS filtrate were assayed to determine their inhibitory activity after size-exclusion chromatography, the inhibitory activity was found in the fractions with a low molecular weight. The inhibitory effect was completely reversed when catalase was added to the cell-free filtrate of AHS. A quantitative method also revealed high production (approximately 3 mmol/l) of hydrogen peroxide in the AHS filtrate with the best inhibitory activity. Electron microscopy of bacteria exposed to AHS filtrate with an inhibitory effect showed changes similar to bacteria exposed to hydrogen peroxide. Results—When fractions of AHS filtrate were assayed to determine their inhibitory activity after size-exclusion chromatography, the inhibitory activity was found in the fractions with a low molecular weight. The inhibitory effect was completely reversed when catalase was added to the cell-free filtrate of AHS. A quantitative method also revealed high production (approximately 3 mmol/l) of hydrogen peroxide in the AHS filtrate with the best inhibitory activity. Electron microscopy of bacteria exposed to AHS filtrate with an inhibitory effect showed changes similar to bacteria exposed to hydrogen peroxide. Conclusions—We conclude that the inhibitory effect of AHS is most likely due to the production of hydrogen peroxide. The significance of hydrogen peroxide production of AHS is discussed in relation to the non-specific and specific mucosal defence systems.

Self-inflicted negative pressure of the external ear canal : a common cause of isolated malleus fractures

We report the investigation of the cause of isolated malleus fractures based on four recent patients at our clinic and five patients reported to us by Swedish otosurgeons. In recent years we have treated four patients with isolated malleus fractures. Colleagues in the Swedish Society of Otosurgeons were encouraged to send us reports on patients with diagnosed isolated malleus fractures, resulting in five more cases. A literature review focusing on the cause and management of this injury was also conducted. Eight of nine patients in the Swedish material had the same history. The patients had inserted a finger into the external auditory canal (most often after a bath) and then pulled it out. Immediately afterwards they experienced a short pain and a hearing loss. An audiogram revealed a conductive hearing loss. After exploration of the middle ear, the most common operation performed was ossiculoplasty. In the present material the most common cause of isolated malleus fractures was a sudden negative pressure in the external auditory canal created by a quick outward movement of a finger in the external ear canal. Since this fracture appears to be infrequent, it can easily be missed at otomicroscopic examination. Tympanometry and pneumatic otomicroscopy are helpful diagnostic tools.

Bacterial adherence to pharyngeal cells: in vitro studies with alpha-haemolytic streptococci and Haemophilus influenzae.

We examined the adherence to pharyngeal cells of alpha-haemolytic Streptococci (AHS) and Haemophilus influenzae , representing normal flora and otitis media (OM) pathogens, respectively. The bacteria were incubated with epithelial cells brushed from the tonsils, adenoid or tubal orifice of children and adults. Adherence varied among the clinical isolates of AHS and H. influenzae . AHS adhered better to epithelial cells from a child compared with those sampled from an adult. The bacteria adhered better to cells from the tubal orifice compared with those sampled from the adenoid. The selective attachment of AHS to certain cells but not to others could not be correlated to apoptotic/necrotic cells versus viable cells. Incubation of epithelial cells with an isolate of AHS with good inhibitory activity against OM pathogens showed almost no adherence of bacteria to the epithelial cells after 12 and 24 h of incubation. If, however, an isolate of AHS with weak inhibitory activity was incubated with the cells, the bacteria that were attached to the epithelial cells from the beginning showed overgrowth in the broth and increasing attachment to the cells after 12 and 24 h. Thus the inhibitory activity of AHS could also affect the adherence of potential pathogens to the mucosal surfaces. The adherence pattern may at least partially explain the difference in susceptibility to OM between children and adults.