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Abstract
Knowledge of the ceruminolytic activity of commercially available ear cleansing products assists the practitioner to choose the best available product for
specific clinical situations. The aim of this study was
to quantify and compare the ceruminolytic activity of
commercially available canine ear cleansers. For this
purpose, the ceruminolytic activity of 13 ear cleansers
was evaluated using a standardized synthetic cerumen (SSC) that mimics the composition and texture of
canine cerumen. The test products were incubated
with mild agitation for 20 min with 500 mg of SSC previously compacted at the bottom of a test tube. Ceruminolytic activity was then assessed by quantifying
the SSC removed by decantation. This procedure was
repeated five consecutive times on each tube simulating repeated applications in the canine ear canal. Good
repeatability among replicates was found in this
assay, allowing direct comparisons between products. The final percentage of SSC elimination ranged
from none (similar to water), between 8 and 39% for
three products and up to 90% for one product
(P < 0.001). It is concluded that, in the experimental
conditions used in this study, only 1/13 products had
significant ceruminolytic activity.
Received 21 July 2005; accepted 21 December 2005
Introduction
Canine ear canals are rich in sebaceous and ceruminous
glands which produce the lipids contained in cerumen.1
Cerumen is a complex mixture of exfoliated cells, waxes,
oils, free fatty acids, esters, immunoglobins and proteins,
which act as an antimicrobial and protective barrier for the
ear canal.1,2 Debris entering the ear canal are trapped and
2006 The Authors. Journal compilation 2006 European Society of Veterinary Dermatology. 17; 121127
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J Snchez-Leal et al.
33.6%
33.6%
9.4%
10.9%
12.5%
sufficient quantities and homogeneity of the material. The composition of this cerumen was based on the average composition of
natural canine cerumen as described in the literature,1,1618 although
concentration of oleic acid was slightly lowered in order to obtain
adequate consistency and malleability. Final composition of SSC is
described in Table 1. Briefly, each component of SSC was carefully
weighed. Solid ingredients (cholesterol, myristic acid and palmitic
acid) were placed in a beaker and blended with a spatula. Then, liquid
ingredients (squalene and oleic acid) were slowly added and thoroughly
mixed until a homogeneous mass was obtained.
In order to assess chemical similarity between SSC and natural
cerumen, canine and human cerumen samples, olive oil, and SSC
were dissolved in chloroform : methanol (2:1) and applied to thin layer
chromatography (TLC) plates. Normal canine cerumen was obtained
from a healthy beagle colony (Isoquimen SL, Spain). Human cerumen
was a pool of samples generously provided by Consejo Superior de
Investigaciones Cientficas (CSIC). Olive oil was included as a reference for its high content in triglycerides. Plates were developed with
an hexane : diethyl ether : acetic acid mixture (80:20:1) and air dried
in a fume cupboard. Spots corresponding to every chemical component of the samples were visualized after spraying the plates with
perchloric acid (40%) and heated at 180 C. Individual components
were identified using in parallel standard solutions of each one. Good
correspondence in the retardation factor (Rf) values of the most
important lipidic fractions was found (Fig. 1).
Aliquots of approximately 500 mg of the SCC mass were rolled and
placed into individual 3 mL polypropylene test tubes (Nirco SA, Spain.
Ref: 17-5594D). The size of the tubes (11 75 mm) was chosen to
simulate the conditions of a dog ear canal. It was estimated that
500 mg of SSC was the maximum amount of cerumen that would still
leave enough tube space for the tested products. The tubes were
placed in a vertical position in a test tube rack and heated at 41 C for
5 min in order for the melted cerumen to homogeneously slide and
to compact to the bottom of the tube thus resembling the conditions
in which cerumen is found in the dog ear canal.
Once cold and solidified, the tubes were carefully weighed and
filled with 2 mL of the test product. This amount was chosen because
it allows good interaction with the SSC plug while is compatible with
the size of the tubes used and with the recommended ear cleansing
practice. Filled tubes were then incubated in a shaker at 35 C for
20 min with mild agitation at 130 cycles/min (Denmark Comfort
Heto Master Shake, Heto-Holten A /S, France. Type: SBD50-1), thus
resembling the temperature,19,20 contact time4 and head shaking4
that would occur in a real ear. Tubes were then inverted for 1 h, allowing
enough time for the dispersed SSC and the test product mixture to
slide out of the tube. Finally, all tubes were weighed again to calculate
the exact amount of remaining SSC. This procedure was repeated five
times on each tube simulating consecutive applications of the products
until complete removal of the SSC plug in the positive control was
achieved (tests 15).
Initial and final weights of the tubes in each test were used to calculate the percentage of SSC removed. Individual results of each replicate were used to calculate the average percentage of removed SSC.
Products tested
The products tested were samples of the most representative commercially available products in several countries. All products were
freshly obtained from authorized distributors of Belgium, Canada,
France, Italy and Spain, and were used well in advance of their expiry
date. Details of each product are described in Table 2.
The trial was run in three phases. In phase I, four products were
chosen [OT (Otoclean, Laboratorios Dr Esteve SA, Spain), CE
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Statistics
Percentages of SSC removed by all products in each test were statistically analysed by analysis of variance (ANOVA) to detect the existence
of any significant difference, whereas Tukey pairwise comparisons test
was used to isolate the differences among treatments within each
test. A level of significance of P < 0.05 was selected. SIGMA STAT version
2.0 Copyright 199295 Jandel Corporation was used for the analyses.
2006 The Authors. Journal compilation 2006 European Society of Veterinary Dermatology.
Declared composition
Details
AUR
CE
CLO
DETE
EP
NET
O-FR
O-LA
OOR
OT
ROU
SP
VET
Figure 2. Appearance of a selection of the tubes after three consecutive assays in phase I showing different degrees of standardized synthetic cerumen (SSC) disintegration. As described in Table 1, after test
3, water (A) and Epiotic (EP) did not show any effect on the SSC plug,
whereas Cerumene (CE), Specicare (SP) and Otoclean (OT) induced
an increasing degree of SSC disintegration and removal.
Results
Figure 2 shows the appearance of the test tubes in several
stages of the plug breakage. A small quantity of the test
products remained inside the tubes even when all of the
SSC was apparently removed; therefore 100% efficacy
could never be achieved. Results of phases I, II and III are
shown in Tables 3, 4 and 5 and represented in Figs 3, 4
and 5, respectively. The negative values indicate impregnation of the product in the SSC without its disintegration
and elimination.
In phase I, the final percentage of SSC elimination was
almost complete for OT (86.5%) and was significantly
greater than SP (23.4%) and CE (8.4%) (P < 0.001). These
Discussion
In this study, only four products were able to remove all or
part of the SSC plug. The other nine products performed
2006 The Authors. Journal compilation 2006 European Society of Veterinary Dermatology.
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J Snchez-Leal et al.
Table 3. Percentage of standardized synthetic cerumen (SSC) removed after each run in phase I. Mean standard deviation (n = 7)
Sample
Test 1
Test 2
A
OT
CE
EP
SP
2.6 0.8
5.1 2.3b,c
2.6 1.1a,c
3.5 0.5a
8.0 1.9c
a
Test 3
3.7 0.8
26.7 5.6b
5.1 1.9c,d
5.7 0.6a
11.0 1.3c
a
Test 4
3.6 0.6
64.6 18.2b
7.7 3.1c
6.2 0.8a
15.8 1.5d
a
Test 5
2.9 0.8
81.9 2.5b
7.8 2.1c
5.5 0.3a
24.1 2.2d
5.3 1.1a
86.5 2.3b
8.4 2.4c
8.1 0.4a
23.4 2.8d
Superscripts with different letter in the same column mean that the products showed statistical differences (P < 0.01) in Tukey pairwise
comparisons test. Negative values indicate that there has been only impregnation of the product in the SSC, but its disintegration and elimination
were not achieved. n = number of replicates; A = water; OT = Otoclean; CE = Cerumene; EP = Epiotic; SP = Specicare.
Table 4. Percentage of standardized synthetic cerumen (SSC) removed after each run in phase II. Mean standard deviation (n = 8)
Sample
Test 1
Test 2
A
OT
VET
ROU
CLO
O-LA
AUR
0.5 0.3
11.7 3.7b
1.5 0.9a
3.6 0.7a
1.4 0.5a
3.8 0.5a
6.5 1.5a
a
Test 3
0.6 0.1
34.5 5.6b
0.1 1.2a
0.2 0.6a
0.2 0.8a
1.6 0.8a
7.9 1.8c
a
Test 4
1.4 0.4
79.0 12.2b
0.7 1.3a
2.4 1.7a
1.6 0.5a
4.5 1.1a
4.5 5.6a
a
Test 5
1.5 0.3
89.4 2.4b
1.1 1.4a,c
3.0 0.9a,c,d
3.0 0.9a,c,d
5.9 1.5a,d
6.5 1.7a,d
a
1.8 0.4a
90.0 2.4b
2.0 2.0a,c
5.6 2.0a
3.3 1.0a,c
9.2 1.6c
6.2 1.5a
Superscripts with different letter in the same column mean that the products showed statistical differences (P < 0.01) in Tukey pairwise
comparisons test. Negative values indicate that there has been only impregnation of the product in the SSC, but its disintegration and elimination
were not achieved. n = number of replicates; A = water; OT = Otoclean; VET = VET (ear cleansing solution), ROU = Routeen, CLO = Clorexyderm,
O-LA = Otolane, AUR = Aurinet.
Table 5. Percentage of standardized synthetic cerumen (SSC) removed after each run in phase III. Mean standard deviation (n = 5)
Sample
Test 1
Test 2
Test 3
Test 4
Test 5
A
OT
O-FR
NET
OOR
DETE
0.8 0.2a
12.2 3.0b
0.8 0.6a
14.1 3.4b
2.7 0.6a
0.7 0.7a
0.6 0.2a
23.1 4.0b
1.4 1.1a
16.9 4.3c
2.0 2.0a
1.4 0.5a
0.7 0.2a
51.2 9.1b
1.2 0.8a
22.7 3.2c
2.2 2.8a
0.3 0.6a
1.2 0.7a
85.0 4.1b
2.4 1.1a
27.9 4.7c
5.5 3.2a
4.6 2.2a
0.9 0.2a
89.6 3.5b
0.7 1.4a
39.1 4.5c
2.1 2.6a
0.7 1.1a
Superscripts with different letter in the same column mean that the products showed statistical differences (P < 0.01) in Tukey pairwise
comparisons test. Negative values indicate that there has been only impregnation of the product in the SSC, but its disintegration and elimination
were not achieved. n = number of replicates; A = water; OT = Otoclean; O-FR = Otifree, NET = Netaural, OOR = Oorreiniger, DETE = Dtcane.
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2006 The Authors. Journal compilation 2006 European Society of Veterinary Dermatology.
no differently from water and only had the effect of a moistening agent. OT was fastest and most effective at breaking up the SSC plug. In all three phases, its effects were
already noticeable and statistically different from water in
the first test and confirmed from then on. OT was able to
break up the entire SSC plug, even thought it did not
achieve 100% efficiency as some product and cerumen
residues remained stuck to the test tube wall and were
not detached by simple gravity.
The ceruminolytic activity of NET, SP and CE was visible
and statistically different from water in the first or second
test and gradually increased in the following extractions,
reaching percentages of 39%, 23% and 8%, respectively.
As in the case of OT, these percentages are probably
lower than the total cerumen removed because some
may be left on the tube wall. As an increasing amount was
removed in each test, it is probable that with more extractions they would have finally achieved complete plug
dispersion as OT. Five consecutive tests were insufficient
to observe any cerumen dispersing effect with EP, O-LA,
AUR, VET, ROU, CLO, O-FR, OOR, DETE or water (A).
The assay showed good repeatability among replicates
with relatively small standard deviations. OT and water,
both tested in all the phases, produced results that were
practically identical throughout. These results confirm that
the method is suitable for direct, objective comparisons
between the products.
Dispersing the lipidic fraction of cerumen facilitates
cleaning of the ear canal.4,6 Authors such as Nielloud
et al.12 have therefore studied the effect of canine ear
cleansers in the dissolution of a synthetic cerumen. These
authors quantified the effect of the cleansers after passing
the mixture of cleanser and cerumen through a 10-m
filter. This method is dissimilar to natural conditions, as it
measures only dissolved earwax, the most extreme way
in which earwax can be removed. In the natural cleansing
process in the canine ear canal, the cleanser would first
impregnate and disaggregate the earwax, thus facilitating its flushing out of the ear canal without necessarily
dissolving the lipids completely. The cerumen used by
Nielloud et al.12 did not include ingredients known to
be constant constituents of canine cerumen such as
Conclusions
Given the high degree of complexity in the formulation of
most commercially available ear cleansers and the limited
information available on their labelling, particularly on its
detailed composition, it is difficult for the practitioner to
have reliable data of their ceruminolytic properties. Moreover, the results obtained in this study indicate that the
ceruminolytic capabilities of the products available on
the market are not easily deduced by their stated composition. Also, notwithstanding the action of their principle
components, the combined action of the complete formulation has to be considered and this information is normally
not in the public domain. The present study was not
designed to elucidate the effects of each of the different
components included in these products; however, the
methods described in this study could be a useful guide
for further research in this field. For the veterinary practitioner, the results of the present trial give objective data on
the comparative ceruminolytic effectiveness of the most
common ear cleansing products on the market. This can
be useful to choose by ceruminolytic capability, the most
appropriate product for each clinical situation. Although
the values obtained apply only to the experimental conditions used in this trial, these results suggest that OT
would be the most effective and rapid ceruminolytic of the
products tested.
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J Snchez-Leal et al.
References
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World Small Animal Veterinary Association, 2001; 27678.
4. Rosychuk RA. Management of otitis externa. Veterinary Clinics of
North America: Small Animal Practice 1994; 24: 92152.
5. Mansfield PD, Steiss JE, Boosinger TR et al. The effects of four,
commercial ceruminolytic agents on the middle ear. Journal of
the American Animal Hospital Association 1997; 33: 47986.
6. Chester DK. Medical management of otitis externa. Veterinary
Clinics of North America: Small Animal Practice 1988; 18: 799
812.
7. Merchant SR. Medically managing chronic otitis externa and
media. Veterinary Medicine 1997; 92: 51734.
8. Lloyd DH, Bond R, Lamport I. Antimicrobial activity in vitro and in
vivo of a canine ear cleanser. Veterinary Record 1998; 143: 1112.
9. Lloyd DH, Lamport AI, Gatto H et al. Potency of two ear cleansers
in vitro against staphylococcus intermedius, Pseudomonas
aeruginosa and Malassezia pachydermatis. In: AVEPA eds. Proceedings of the Twenty-Seventh Annual Meeting of the World
Small Animal Veterinary Association. Granada: World Small
Animal Veterinary Association, 2002; 1: 33.
10. Roura X, Sabat D, Homedes J. Efficacy study of an otic solution
(Otoclean) in the cleaning of the ear canal in dogs. In: AVEPA
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Rsum La connaissance de laction crumenolytique des nettoyants auriculaires aide le clinicien choisir
le meilleur produit en fonction de la situation clinique. Le but de cette tude tait de quantifier et de comparer
laction crumenolytique de produits commercialiss. Dans ce but, lactivit crumenolytique de 13
nettoyants a t value en utilisant un crumen synthtique (SSC) qui imite la composition et la texture
du crumen du chien. Les produits tests ont t incubs avec une agitation modre pendant 20 minutes
avec 500 mg de SSC pralablement compact la base dun tube essai. Lactivit crumenolytique a t
dtermines en quantifiant la quantit de SCC limine aprs dcantation. Cette procdure a t rpte
5 fois conscutivement dans chaque tube, imitant des applications rptes dans le conduit auditif externe.
Une bonne rptabilit a t observe, permettant une comparaison des produits tests. Le pourcentage
final dlimination du SCC variait de nul (similaire de leau), entre 8 et 39% pour 3 produits, et jusqu 90%
pour un produit (P < 0.001). Il est conclu que dans les conditions exprimentales de cet essai, seul 1/13
produit prsente une activit crumenolytique significative.
Resumen El conocimiento de la actividad cerumenoltica de soluciones de limpieza comerciales ticas
ayuda al clnico a escoger el mejor producto disponible para diferentes situaciones clnicas. El propsito de
este estudio fue cuantificar y comparar la actividad cerumenoltica de varios productos ticos de limpieza
para perros. Con este objetivo, 13 productos limpiadores fueron evaluados utilizando un cerumen sinttico
modelo que mimetiza la composicin y textura del cerumen canino. Los productos probados fueron incubados
con ligera agitacin durante 20 minutos con 500 mg del cerumen sinttico previamente compactado en el
fondo de un tubo de ensayo. La actividad cerumenoltica se evalu mediante la cuantificacin del cerumen
sinttico removido mediante decantacin. Este procedimiento fue repetido en cinco ocasiones en cada tubo,
simulando as aplicaciones repetidas en el odo canino. Se encontr bastante consistencia en las repeticiones, lo que permiti una comparacin entre los diferentes productos. El porcentaje final de cerumen
sinttico eliminado vario entre 0 (similar a agua), de 839% para tres productos, y hasta un 90% para un
producto (P < 0.001). Concluimos que bajo estas condiciones experimentales, solo uno de 13 productos
present actividad cerumenoltica suficiente.
Zusammenfassung Das Wissen ber die cerumenolytische Aktivitt von kommerziell erwerblichen
Ohrreinigungsprodukten hilft dem Praktiker das bestmgliche Produkt fr spezielle klinische Situationen zu
whlen. Das Ziel dieser Studie war es, die cerumenolytische Aktivitt von kommerziell erwerblichen
Ohrreinigern fr Hunde quantitativ zu bestimmen und zu vergleichen. Zu diesem Zweck wurde die cerumenolytische Aktivitt von 13 Ohrreinigern an einem standardisierten synthetischen Cerumen (SSC),
126
2006 The Authors. Journal compilation 2006 European Society of Veterinary Dermatology.
welches in Zusammensetzung und Textur dem Hundecerumen gleicht, evaluiert. Die zu testenden Produkte
wurden bei leichter Bewegung fr 20 Minuten mit 500mg SCC inkubiert, welches davor am Boden des
Reagenzrhrchens zusammengepresst worden war. Die cerumenolytische Aktivitt wurde dann beurteilt,
indem die durch Dekantieren entfernte Menge an SCC bestimmt wurde. Diese Vorgangsweise wurde
fnfmal hintereinander bei jedem Reagenzrhrchen wiederholt, um die wiederholte Applikation in den
Ohrkanal zu simulieren. Bei diesem Test wurde eine gute Reproduzierbarkeit zwischen den Replikaten
gefunden, was einen direkten Vergleich der Produkte erlaubte. Der Restprozentsatz fr die Eliminierung von
SCC erstreckte sich von 0 (hnlich wie Wasser), zwischen 8 und 39% fr drei Produkte und bis zu 90% fr
ein Produkt (P < 0.001). Es wird daraus geschlossen, dass unter den experimentellen Bedingungen, wie
sie in dieser Studie verwendet worden waren, nur 1/13 Produkten eine signifikante cerumenolytische
Aktivitt aufwies.
2006 The Authors. Journal compilation 2006 European Society of Veterinary Dermatology.
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