215 DISC

Veterinary Dermatology 2001, 12, 4147

Rapid identication of tissue micro-organisms in skin

biopsy specimens from domestic animals using
polyclonal BCG antibody
TERRI E. BONENBERGER,*} PETER J. IHRKE,{} DIANE K. NAYDAN{ and VERENA
K. AFFOLTER{}
*Veterinary Medical Teaching Hospital
{Department of Medicine and Epidemiology, {Department of Pathology, Microbiology and Immunology,
School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
}Davis Dermatology Group, Veterinary Medical Teaching Hospital, University of California, Davis, CA
95616, USA
(Received 14 December 1999; accepted 24 February 2000)

Abstract Immunostaining with polyclonal anti-Mycobacterium bovis (BCG) was evaluated as a single screening method for the histological identication of micro-organisms in skin biopsy specimens from various
veterinary species. Conrmed archival cases infected with Mycobacteria, Nocardia, Actinobacillus,
Actinomyces, Streptococcus/Staphylococcus, Dermatophilus, spirochetes, Blastomyces, Coccidioides, Cryptococcus, Histoplasma, dermatophytes, Malassezia, Sporothrix, Leishmania, Pythium, phaeohyphomycetes and
Prototheca organisms were selected. A total of 70 skin biopsy specimens from the dog, cat, horse, ox and llama
were evaluated. The anti-BCG immunostain labelled bacteria and fungi with high sensitivity and minimal
background staining but did not label spirochetes and protozoa (Leishmania). Dierences were not noted
between veterinary species. The results indicate that immunostaining with polyclonal anti-BCG is a suitable
screening technique for the rapid identication of most common bacterial and fungal organisms in paranembedded specimens. Also, mycobacterial and nocardial organisms were identied more readily with the antiBCG immunostain in comparison to the histochemical stains.

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Keywords: BCG, immunohistochemistry, Mycobacteria, Nocardia, pathology-veterinary, polyclonal antibody.

INTRODUCTION
Currently, the identication of various infectious
agents in histological specimens requires the application of multiple special stains. The histochemical
stains routinely used for the identication of bacteria,
fungi and protozoa (Leishmania) include Fite-Faraco,
Ziehl-Neelsen, Brown-Brenn (B & B), Giemsa,
periodic acid-Schi (PAS), Gomori's methenamine
silver (GMS), mucicarmine and Warthin-Starry.1 The
application of multiple special stains to skin biopsy
specimens is time consuming and costly. In routine
dermatopathology, as well as other areas of pathology, there is a growing need for a single, rapid, costeective and highly sensitive screening tool for the
detection of micro-organisms.
Bacille Calmette-Guerin (BCG) is an attenuated
strain of Mycobacterium bovis. Previous studies in
human medicine have demonstrated the cross-reactivity of polyclonal anti-Mycobacterium bovis (BCG)

Correspondence: Dr Peter Ihrke, Department of Medicine and

Epidemiology, School of Veterinary Medicine, University of
California, Davis, CA 95616, USA.
This project was partially funded by the George H. Muller Fund
for Research in Veterinary Dermatology.
# 2001 Blackwell Science Ltd

Ref start

antibodies to a broad spectrum of fungi and

bacteria.25 Kutzner and colleagues described the
anti-BCG immunostaining technique as a promising
screening tool for the detection of common infectious
micro-organisms in humans.2 The purpose of this
study was to evaluate if immunostaining with
polyclonal anti-BCG antibody was an appropriate
diagnostic tool for veterinary medicine.

MATERIALS AND METHODS

Tissue preparation
Skin biopsy specimens with conrmed intralesional
micro-organisms common to veterinary medicine
were selected.6,7 Archival cases with cutaneous
Mycobacteria (n = 7), Nocardia (n = 5), Actinobacillus (n = 5), Actinomyces (n = 5), Streptococcus/Staphylococcus
(n = 5),
Dermatophilus
(n = 5),
spirochetes (n = 2), Blastomyces (n = 2), Coccidioides
(n = 6), Cryptococcus (n = 5), Histoplasma (n = 2),
dermatophytes (n = 7), Malassezia (n = 5), Sporothrix (n = 3), Leishmania (n = 3), Pythium (n = 1),
phaeohyphomycetes (n = 1) and Prototheca (n = 1)
organisms were selected. One to seven samples of
each infectious disease were chosen. The number of
41

215 DISC
42

T.E. Bonenberger et al.

cases selected from each infectious disease group

depended on availability of archival samples. A total
of 70 skin biopsy specimens were evaluated from the
medical record database at the University of California, Davis, Veterinary Medical Teaching Hospital
from the years 198498. Additional samples were
provided by the Colleges of Veterinary Medicine at
North Carolina State University (two cutaneous
blastomycosis, one cutaneous pythiosis), Louisiana
State University (one cutaneous histoplasmosis) and
the University of Tennessee, Knoxville (one cutaneous histoplasmosis). The following species were
included: dog, cat, horse, ox and llama. Four
micrometer serial sections of formalin-xed, parafn-embedded tissue were obtained. Sections of each
specimen were stained with the polyclonal anti-BCG
immunostain and the appropriate histochemical
stains for comparison purposes. All slides were
reviewed by authors Bonenberger and Aolter and
subsequently divided into three categories based on
staining intensity.
Antigen retrieval and immunohistochemistry
Mounted tissue sections were deparanized in xylene
and then hydrated up to 70% alcohol. Quenching of
endogenous peroxidase was performed by slide
immersion in 0.3% H2O2/methanol for 30 min at
room temperature (258C). Slides were rinsed with
PBS buer/Tween 20. In previous studies, heatinduced epitope retrieval was combined with enzymatic epitope retrieval.2 A pilot study was conducted
to evaluate various antigen retrieval techniques.2,8,9
The antigen retrieval technique was modied from
previous anti-BCG immunostaining techniques to
minimize tissue destruction and maximize contrasting
staining characteristics. Enzymatic retrieval techniques evaluated included pretreatment with proteinase
K (DAKO Corp., Carpinteria, CA, USA), 0.1%
trypsin (Sigma #T7409, Sigma, St Louis, MO, USA)
and 0.4% pepsin (Sigma #P7012). Heat-induced
antigen retrieval techniques evaluated included
steaming, microwaving and autoclaving. Combinations of enzymatic and heat-induced antigen retrieval
techniques were also examined. In this study, antigen
retrieval was achieved by slide immersion in 0.4%
pepsin in pH 3.3 acidulated water for 15 min at 378C.
After antigen retrieval, the 10% normal goat serum
was applied for 20 min at 258C to block nonspecic
binding. The slides were stained with the DAKO
Autostainer (DAKO Corp.). Polyclonal anti-BCG
antibody (DAKO Corp.) was applied at a dilution of
1 : 4000. The slides were incubated for 2 h at room
temperature on the autostainer. After several rinses
with PBS buer/Tween 20, the biotinylated secondary antibody (goat antirabbit IgG (Vector Laboratories, Inc., Burlingame, CA, USA) was applied at a
dilution of 1 : 500 for 30 min at room temperature.
The streptavidin-horseradish peroxidase conjugate
(Vector Laboratories, Inc.) provided the enzyme
label and was applied at a dilution of 1 : 500 for 30
# 2001 Blackwell Science Ltd, Veterinary Dermatology, 12, 4147

min at room temperature. Three-amino-9-ethyl carbazole (AEC) was used as the chromogen and
prepared as directed (Zymed Laboratories, San
Francisco, CA, USA). The sections were counterstained with Mayer's haematoxylin (Sigma). Negative
controls were prepared by omission of the primary
antibody and substitution with either PBS or normal
rabbit serum (diluted to 1 : 4000). Sections of bovine
intestine infected with Mycobacterium paratuberculosis were used as positive controls. Negative controls
were run for each individual tissue sample and a
positive control was run with each batch of samples.
Histochemistry
Appropriate special stains including Fite-Faraco,
Ziehl-Neelsen, Brown-Brenn (B & B), Giemsa,
periodic acid-Schi (PAS), Gomori's methenamine
silver (GMS), mucicarmine and Warthin-Starry were
performed for each infectious disease based on
recommendations in the literature.1 All stains were
performed according to established procedures.1
RESULTS
The anti-BCG immunostain labelled all organisms
evaluated with the exception of spirochetes and
protozoa (Leishmania). Dierences in staining were
not detected between organisms evaluated in the
domestic animal species represented.
The antigen retrieval technique of proteolytic
digestion with 0.4% pepsin enhanced contrasting
staining characteristics, while minimizing tissue destruction. Combining enzymatic and heat-induced
antigen retrieval techniques produced greater tissue
destruction than proteolytic digestion alone. Many of
the heat-induced antigen retrieval techniques, specically autoclaving and microwaving, also resulted in
signicant tissue destruction. Antigen retrieval with
0.4% pepsin consistently produced optimum results
with excellent tissue preservation and minimal background staining. Histological changes characteristic
of the disease process could be evaluated in addition
to viewing the presence of micro-organisms.
The intensity of the staining with the anti-BCG
immunostain varied between dierent organisms.
Based on these results, the samples were divided into
three categories according to intensity of staining and
therefore ease of organism identication. The categories represented samples in which the anti-BCG
immunostaining was superior (Group 1), equal
(Group 2) or inferior (Group 3) when compared to
the appropriate histochemical stains.
The anti-BCG immunostain was superior in labelling mycobacterial and nocardial organisms (Fig.
1a,b). In both cases, these micro-organisms were
more easily identied when compared to the FiteFaraco (Mycobacteria, Nocardia) and Ziehl-Neelsen
(Mycobacteria) stains. The anti-BCG immunostain
labelled whole and fragmented organisms located

215 DISC
Detection of pathogens using polyclonal BCG antibody
Figure 1. Feline opportunistic cutaneous
mycobacteriosis:(a) anti-BCG
immunostaining of intracytoplasmic
mycobacterial organisms located within
macrophages (arrows) (6540); (b)ZiehlNeelsen histochemical staining of
intracytoplasmic mycobacterial organisms
located within a macrophage (arrow)
(6540).

43

(a)

(b)

intra and extracellularly. In most cases, the organisms

could be recognized with the anti-BCG immunostain
at lower microscopic powers. The comparative
histochemical stains were superior to the anti-BCG
immunostain for identication of Cryptococcus,
Pythium and Prototheca micro-organisms (Fig.
2a,b). In these samples, the anti-BCG immunostain
labelled only part of the micro-organism capsule or
hyphal wall, whereas the histochemical stain evenly
stained all organisms. The anti-BCG immunostain
did not label any specimens containing spirochete or
Leishmania organisms. The anti-BCG immunostain
was equal in ecacy to histochemical stains for
identication of all remaining organisms (Fig. 3a,b).
DISCUSSION
The results of this study indicate that the anti-BCG
immunostaining technique was an eective tool for
the identication of common bacterial and fungal

organisms within skin biopsy specimens from multiple species of domestic animals. The anti-BCG
immunostain labelled bacteria and fungi with high
sensitivity and minimal background staining, but did
not react with spirochetes or protozoa (Leishmania).
The human literature reports that the anti-BCG
immunostain is ineective at labelling spirochetes,
protozoal or viral organisms.2 Therefore, these results
corroborate previous studies in human medicine,2
although viral-infected tissues were not examined.
Previous uses of BCG in veterinary medicine have
included utilization as a vaccine, an adjuvant and as a
biological response modier in the treatment of
certain canine neoplasias and sarcoids in horses.10
The development of BCG as an immunostain began
when pathologists noted that BCG fragments persisted at inoculation sites beyond when the bacilli
could be detected by special histochemical stains.11,12
The anti-BCG immunostain was then used to detect
Mycobacterium leprae in nerve biopsies.12 The antiBCG immunostaining technique was reported to be
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215 DISC
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T.E. Bonenberger et al.

(a)

Figure 2. Feline cutaneous cryptococcosis:

(a) anti-BCG immunostaining of
cryptococcal organisms. Varying degrees of
stain uptake present. Note organism with
positive stain uptake (small arrow) and
negative stain uptake (large arrow) (6540);
(b) mucicarmine histochemical staining of
cryptococcal organisms. Note even staining
of all cryptococcal organisms (arrow) (6540).

(b)

more sensitive in detection of M. leprae than

histochemical stains (Fite-Faraco, Kinyoun stains)
because the authors identied cases in which the
organism was cultured and labelled with the antiBCG immunostaining technique, but was not identied by histochemical stains.10 Subsequently polyclonal
antibodies
to
Mycobacterium
bovis,
Mycobacterium duvalii and Mycobacterium paratuberculosis were evaluated as immunostains for the
detection of other mycobacterial species within tissue
specimens. Results suggesting a higher sensitivity for
detection of mycobacterial organisms with the antiBCG immunostain when compared to routine
histochemical stains were reported.13
Cross-reactivity between antimycobacterial immunostains and fungi was noted when biopsy specimens
containing fungal organisms were used as failed
negative controls.3 This led to the concept of using
anti-BCG antibody as a broad-based screening tool
for the identication of bacterial and fungal organisms within tissue sections. The human literature
reported uniform labelling of pathogens including
# 2001 Blackwell Science Ltd, Veterinary Dermatology, 12, 4147

most bacterial and fungal organisms with the exception of spirochetes, protozoa (Leishmania) and viruses.
The cross-reactivity suggests a shared antigenicity
between dierent infectious agents.35 The theory that
species conservation results in conservation of specic
cell wall components and that these antigenic components may function in assisting the organism to
parasitize the host has been suggested.3 This shared
antigenicity can be used to identify various infectious
organisms. Therefore, the anti-BCG immunostaining
technique was considered a valid screening tool for
identication of most bacterial and fungal microorganisms in tissue sections.2
In this study, mycobacterial and nocardial organisms were easier to identify with the anti-BCG immunostain when compared with serial sections stained
with Fite-Faraco and Ziehl-Neelsen histochemical
stains. These results are consistent with previous
studies and case reports in human medicine which
have suggested that the anti-BCG immunostaining
technique is more sensitive than histochemical stains
for the detection of mycobacteria and nocardia.12,1417

215 DISC
Detection of pathogens using polyclonal BCG antibody
Figure 3. Canine dermatophytosis: (a)
anti-BCG immunostaining of
dermatophyte hyphae. Bright, even
staining of all fungal hyphae. Note
phagocytosed fungal material present
within macrophage (arrow) (6540);
(b)GMS histochemical staining of
dermatophyte hyphae. Bright, even
staining of all fungal hyphae (6540).

45

(a)

(b)

Immunohistochemical staining techniques can be

more sensitive because they detect mycobacterial
antigens in tissue sections by recognizing whole
organisms and partial fragments.12,1416 Increased
sensitivity of immunostaining over special stains
could not be evaluated because the authors did not
have any available cases where mycobacterial and
nocardial organisms were identied with cultures but
were not identied with histochemical stains in
biopsy specimens.
The anti-BCG immunostain may be particularly
helpful in diagnosing cases of opportunistic mycobacterial infections. These organisms often are
present in extremely low numbers within tissue and
may be dicult to visualize within skin biopsy
specimens despite the use of special stains.6,10,18
Therefore, an immunostaining technique that detects
antigenic fragments would be especially helpful in
cases where only remnants of organisms are present
or organisms are present in low numbers. Further

studies are needed to determine if the anti-BCG

immunostaining technique is a more sensitive tool for
the identication of mycobacteria and nocardia than
routine histochemical stains in veterinary medicine.
Although there were micro-organisms (Cryptococcus, Pythium and Prototheca) in which the anti-BCG
immunostain failed to evenly label the capsule or
hyphal walls, there was some stain uptake by the
micro-organisms in all cases. Uneven staining implies
that the cell walls of organisms are not uniformly
binding to the antibody.8,9 This may be the result of a
transient display of similar antigenic determinants
that are recognized only during certain phases of
maturation or are rapidly modied and become
unrecognizable by the anti-BCG antibody.3 Alternatively, the length of formalin xation prior to
processing may interfere with the ability of the
antibody to label the micro-organism.8,9 Studies have
reported that for optimal immunostaining, tissue
xation should not exceed 48 h.19 Tissue stored in
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215 DISC
46

T.E. Bonenberger et al.

formalin xative for longer periods generally has

decreased staining intensity due to the loss of cellular
antigens.19 Formalin is a cross-linking xative producing links between reactive end-groups of adjacent
protein chains.8 Fixed proteins can retain their
antigenicity only if the cross-linking does not aect
the epitope recognized by the antibody.8 Comparison
of formalin-xed tissue with fresh frozen tissues would
be required to determine if xation is contributing to
the uneven staining observed. Since this study
evaluated archival tissue specimens, it was not possible
to obtain fresh-frozen tissue samples for comparison.
In conclusion, polyclonal anti-Mycobacterium bovis (BCG) is a suitable screening technique for the
identication of most common bacterial and fungal
organisms in paran-embedded skin biopsy specimens from a variety of domestic animals. Additional
studies are required to determine if the anti-BCG
immunostaining technique is a more sensitive tool to
identify mycobacterial and nocardial organisms.

ACKNOWLEDGEMENTS
The authors would like to thank Drs T. Olivry, College
of Veterinary Medicine, North Carolina State University; C. Foil, College of Veterinary Medicine,
Louisiana State University; and L. Schmeitzel, College
of Veterinary Medicine, University of Tennessee, for
providing tissue specimens representative of skin
diseases uncommonly seen in California.

REFERENCES
1. Sheehan, D.C., Hrapchak, B.B., eds. In: Theory and
Practice of Histotechnology. 2nd edn. Columbus, OH:
Battelle Press, 1987: pp 99, 137251.
2. Kutzner, H., Argenyi, Z.B., Requena, L., Rutten, A.,
Hugel, H. A new application of BCG antibody for
rapid screening of various tissue microorganisms.
Journal of American Academy of Dermatology 1998;
38: 5660.
3. Wiley, E.L., Beck, B., Freeman, R.G. Reactivity of
fungal organisms in tissue sections using antimycobacterial antibodies. Journal of Cutaneous
Pathology 1991; 18: 2049.
4. Thorns, C.J., Morris, J.A. Shared epitopes between
mycobacteria and other microorganisms. Research in
Veterinary Science 1986; 41: 2756.
5. Daniel, T.M., Janicki, B.W. Mycobacterial antigens: a
review of their isolation, chemistry, and immunological
properties. Microbiological Reviews 1978; 42: 84113.

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6. Scott, D.W., Miller, W.H., Grin, C.E., eds. Muller

and Kirk's Small Animal Dermatology. 5th edn.
Philadelphia: W.B. Saunders, 1995: pp 279392.
7. Scott, D.W. Large Animal Dermatology. Philadelphia:
W.B. Saunders, 1988: pp 120202.
8. Polak, J.M., Van Noorden, S. Introduction to
Immunocytochemistry. 2nd edn. New York: SpringerVerlag, 1997.
9. Haines, D.M., Chelack, B.J. Technical considerations
for developing enzyme immunohistochemical staining
procedures on formalin-xed paran-embedded tissues
for diagnostic pathology. Veterinary Diagnostic
Investigations 1991; 3: 10112.
10. Greene, C.E., ed. Infectious Diseases of the Dog and
Cat. 2nd edn. Philadelphia: W.B. Saunders, 1998: pp
303325, 878.
11. Higuchi, S., Moritaka, S., Dannenberg, A.M., Jr,
Aronti, L.F., Azuma, I., Daniel, T.M. et al. Persistence of protein, carbohydrate and wax components of
tubercle bacilli in dermal BCG lesions. American
Review of Respiratory Diseases 1981; 123: 397401.
12. Mshana, R.N., Humber, D.P., Harboe, M., Belehu, A.
Demonstration of mycobacterial antigens in nerve
biopsies from leprosy patients using peroxidaseantiperoxidase immunoenzyme technique. Clinical
Immunology and Immunopathology 1983; 29: 35968.
13. Wiley, E.L., Mulhollan, T.J., Beck, B., Tyndall, J.A.,
Freeman, R.G. Polyclonal antibodies raised against
Bacillus Calmette-Guerin, Mycobacterium duvalii, and
Mycobacterium paratuberculosis used to detect
Mycobacteria
in
tissue
with
the
use
of
immunohistochemical techniques. American Journal of
Clinical Pathology 1990; 34: 30712.
14. Glover, R.A., Cockerell, C.J. Demonstration of
Nocardia asteroides in tissue using polyclonal antimycobacterial antibodies. Journal of Cutaneous
Pathology 1991; 17: 294.
15. Massone, A.R., Martin, A.A., Ibargoyen, G.S.,
Gimeno, E.J. Immunohistochemical methods for the
visualization of Mycobacterium paratuberculosis in
bovine tissues. Journal of Veterinary Medicine 1990;
37: 2513.
16. Humphrey, D.M., Weiner, M.H. Mycobacterial antigen detection by immunohistochemistry in pulmonary
tuberculosis. Human Pathology 1987; 18: 7018.
17. Plante, Y., Remenda, B.W., Chelack, B.J., Haines,
D.M. Detection of Mycobacterium paratuberculosis in
formalin-xed paran-embedded tissues by the
polymerase chain reaction. Canadian Journal of
Veterinary Research 1996; 60: 11520.
18. Gross, T.L., Ihrke, P.J., Walder, E.J. Veterinary
Dermatopathology. 1st edn. St Louis, MO: Mosby
Year Book, 1992: pp 16389.
19. Rickert, R.R., Maliniak, R.M. Intralaboratory Quality
Assurance of Immunohistochemical Procedures.
Archives of Pathology and Laboratory Methods 1989;
113: 6739.

215 DISC
Detection of pathogens using polyclonal BCG antibody

47

Resume Des immunomarquages avec des anticorps polyclonaux anti-Mycobacterium bovis (BCG) ont ete
evalues comme technique diagnostique pour l'identication histologique des micro-organismes dans des
biopsies cutanees obtenues a partir de diverses especes animales. Des cas conrmes d'infection par
Mycobacteria, Nocardia, Actinobacillus, Actinomyces, Streptococcus/Staphylococcus, Dermatophilus, des
spirochetes, Blastomyces, Coccidioides, Cryptococcus, Histoplasma, des dermatophytes, Malassezia,
Sporothrix, Leishmania, Pythium, des phaeohyphomycetes et Prototheca ont ete etudies. Un total de 70
biopsies (chien, chat, cheval, vache et lama) a ete evalue. Le marquage anti-BCG a colore les bacteries et les
champignons avec une sensibilite elevee et un bruit de fond minime, mais il ne marquait pas les spirochetes et
les protozoaires (Leishmania). Aucune dierence n'a ete observee selon les especes etudiees. Ces resultats
indiquent que l'immunmorquage avec un anti-BCG polyclonal est une technique ecace pour l'identication
rapide de la plupart des agents bacteriens et fongiques dans les biopsies paranees. En outre, les
mycobacteries et les Nocardia ont ete plus facilement mises en evidence avec cette technique que par les
colorations histochimiques. [Bonenberger, T. E., Ihrke, P. J., Naydan, D. K., et Aolter, V. K. Rapid
identication of tissue micro-organisms in skin biopsy specimens from domestic animals using polyclonal
BCG antibody. (Identication rapide des micro-organismes dans les biopsies cutanees d'animaux domestiques
a l'aide d'anticorps polyclonaux BCG.) Veterinary Dermatology 2001; 12: 4147.]
Resumen Se evaluo la inmunotincion con un anticuerpo policlonal anti-Mycobacterium bovis (BCG) como
unico metodo de estudio de la identicacion histologica de microorganismos en biopsias cutaneas de varias
especies domesticas. Se seleccionaron casos de archivo con infeccion conrmada de Mycobacteria, Nocardia,
Actinobacillus, Actinomyces, Streptococcus/Staphylococcus, Dermatophilus, espiroquetas, Blastomyces,
Coccidioides, Cryptococcus, Histoplasma, dermatotos, Malassezia, Sporothrix, Leishmania, Pythium,
feohifomicetos y organismos de Prototheca. Se evaluaron un total de 70 biopsias de perros, gatos, caballos,
bovinos y llamas. La inmunotincion anti-BCG marco bacterias y hongos con alta sensibilidad y un m nimo de
reaccion de fondo pero no marco las espiroquetas y protozoos (Leishmania). No se notaron diferencias entre
especies domesticas. Los resultados indican que la inmunotincion con anticuerpos policlonales anti-BCG es
una tecnica de evaluacion adecuada para la identicacion rapida en tejidos incluidos en parana de los
organismos bacterianos y fungicos mas frecuentes. Ademas, las micobacterias y las nocardias fueron
identicadas con mayor facilidad con la inmunotincion anti-BCG comparado con las tinciones histoqu micas.
[Bonenberger, T. E., Ihrke, P. J., Naydan, D. K., y Aolter, V. K. Rapid identication of tissue microorganisms in skin biopsy specimens from domestic animals using polyclonal BCG antibody. (Identicacion
rapida de microorganismos en tejidos de biopsias cutaneas de animales domesticos utilizando un anticuerpo
policlonal de BCG.) Veterinary Dermatology 2001; 12: 4147.]
Zusammenfassung Immunfarbung mit polyklonalen anti-Mycobacterium bovis (BCG) Antikorpern wurde
als Untersuchungsmethode fur histologische Identizierung von Mikroorganismen in Hautbiopsieproben
verschiedener Tierarten bewertet. Mit Mycobacteria, Nocardia, Actinobacillus, Actinomyces, Streptococcus /
Staphylococcus, Dermatophilus, Spirochaten, Blastomyces, Coccidioides, Cryptococcus, Histoplasma,
Dermatophyten, Malassezia, Sporothrix, Leishmania, Pythium, Phaeohyphomyceten und ProtothecaOrganismen nachgewiesenermassen inzierte Proben wurden ausgewahlt. Siebzig Hautbiopsieproben von
Hund, Katze, Pferd, Rind und Llama wurden ausgewertet. Die anti-BCG-Immunfarbung identizierte
Bakterien und Pilze mit hoher Sensitivitat und minimaler Hintergrundfarbung, farbte aber keine Spirochaten
und Protozoen (Leishmania). Zwischen einzelnen Tierarten wurden keine Unterschiede gesehen. Die Resultate
deuten darauf hin, dass eine Immunfarbung mit polyklonalen anti-BCG Antikorpern ein nutzlicher
Screeningtest fur die schnelle Identizierung der haugsten Bakterien und Pilze bei in Paran eingebetteten
Proben ist. Mykobakterien und Nokardien wurden im Vergleich zu immunhistochemischen Farbungen mit
der anti-BCG Immunfarbung leichter angefarbt. [Bonenberger, T. E., Ihrke, P. J., Naydan, D. K. und
Aolter, V. K. Rapid identication of tissue micro-organisms in skin biopsy specimens from domestic animals
using polyclonal BCG antibody. (Schnellidentikation von Gewebsmikroorganismen in Hautbiopsieproben
von Haustieren mit polyklonalen BCG Antikorpern.) Veterinary Dermatology 2001; 12: 4147.]

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