Isolation and Antibiogram of Aerobic Nasal Bacterial Flora of Apparently Healthy West African Dwarf Goats Keywords Summary
Goats are important in the livestock economy by their adaptability to adverse
flora – Antibiotic – Drug resistance –
environmental conditions as they are good sources of protein and income for
THOLOGIE INFECTIEUSE
the rural poor. Studies conducted on the bacterial flora of the respiratory tract
in goats focused on the pneumonic lungs, with fewer studies on the appa-
rently normal nasal passage and antibiogram of isolated organisms. This study was carried out on 60 apparently healthy West African Dwarf goats. The nasal swab from each goat was analyzed using standard methods. The disc diffu-sion technique was used for the antibiotic sensitivity test. Three hundred and twenty-eight isolates were obtained. The most frequently isolated species was Streptococcus spp., while Escherichia coli and Staphylococcus aureus were the second dominant bacteria. Other species were isolated at relatively lower rates. The isolation of Mannheimia haemolytica and Pasteurella multocida from the nasal cavity of apparently healthy goats in this study reflects their possible role in most common respiratory diseases encountered in small rumi-nants. Most of the bacteria were found to be susceptible to streptomycin, qui-nolones (perfloxacin, ciprofloxacin and ofloxacin) and gentamicin, while they were resistant to tetracycline, augmentin and erythromycin. This study shows the relationship between misuse or unrestricted use of antibiotics and drug resistance. Therefore, there is a need for practitioners and researchers to be informed of the appropriate antibiotics to be used in respiratory infections and during control programs.
ruminants (PPR), contagious caprine pleuro-pneumonia (CCPP)
and pasteurellosis] cause substantial loss through high morbidity
Goats have gained importance in the Nigerian livestock economy as
and mortality. Bacterial pneumopathies are commonly attributed to
a result of their remarkable adaptability to adverse environmental
Mannheimia haemolytica which causes severe damage to the lung.
conditions (19). They are good sources of protein and income for the
In addition, bacterial agents such as Actinomyces pyogenes also
rural poor, especially women and children (14), and a good source
inflict damage on pulmonary tissues in goats (4).
of foreign exchange earnings. Hence, increase in goat production is
It is increasingly difficult to make an etiological diagnosis in
needed to maintain food security and increase earnings (10).
most infectious pneumopathies as a viral agent may be a primary
In spite of the fact that every household keeps goats, their produc-
invader, and, when the local resistance of respiratory mucosas is
tion is not well developed because of factors such as inadequate
lowered, bacterial agents growing in the nose and throat develop
nutrition, poor management and prevailing diseases. Of all the dis-
downward producing multiple bacterial infections. Most of the
eases of goats, those affecting the respiratory tract [peste des petits
infectious agents that cause respiratory diseases are usually normal
Studies conducted on the bacterial flora of the respiratory tract of
1. Department of Veterinary Pathology, University of Ibadan, Nigeria.
domestic animals in various parts of the world mainly focus on the
2. Department of Virology, University of Ibadan, Nigeria. * Corresponding author
pneumonic lungs of sheep and goats (1, 13) with fewer studies on
Tel: +234 80 66 48 60 80; E-mail: [email protected]
the microbial flora of the apparently normal nasal passage (17) and
Aerobic Nasal Bacterial Flora of Goats
on the antibiogram of isolated organisms (2, 11). This study aimed
light microscope (x100). Mixed colonies and Gram negative bacte-
at isolating and characterizing bacteria from the nasal passageways
ria were subcultured on both blood and McConkey agars and further
of apparently healthy West African Dwarf goats. The results of the
incubated aerobically for 24 h. Pure culture of single colony type
antibiotic sensitivity of the isolates are also presented.
from both blood and McConkey agars were transferred onto nutrient
agar slants for a series of biochemical tests including catalase, oxi-
dase and fermentative/oxidative tests for final identification follow-
ing standard procedures (21). For the antibiotic sensitivity test, the
disc diffusion technique was used and inhibition observed as clear
Animals
zones around the antibiotics. Inhibition zones were measured and
The study was carried out from November 2005 through March
measurements greater than 0.5 cm were regarded as susceptible.
2007 on 60 apparently healthy goats purposely selected for experi-
ments. These animals were bought in a batch of 20 goats from indi-
Data analysis
viduals and households in and around Ibadan. They were kept in
the small ruminant pens of the Veterinary Pathology Department,
Descriptive statistics were used to summarize the data generated by
in the experimental animal unit of the Faculty of Veterinary Medi-
the study. The relative abundance of each species and genus were
cine at the University of Ibadan. They were not allowed to graze.
expressed as a percentage of the total number of isolates.
They were fed with concentrates and water was provided ad libi-tum. The animals were less than one year old and were apparently
THOLOGIE INFECTIEUSE Sample collection
Three hundred and twenty-eight bacterial isolates were obtained.
Table I shows the bacteria isolated from the nasal passages of goats
The nasal samples were collected a day after arrival by inserting
and Table II shows the results of the antibiotic sensitivity tests. The
sterile cotton-tipped applicator sticks or swab into the nasal pas-
most frequently isolated species was Streptococcus pyogenes, while
sage after proper cleaning and disinfection of the external wares.
Escherichia coli and Staphylococcus aureus were the second domi-
Each nasal swab was carefully cut and put into a labeled bottle
nant bacteria isolated. Mannheimia haemolytica and Pasteurella
containing 2 mL brain heart infusion broth. The swabs were trans-
multocida were the common important respiratory pathogens of
ported in a cool box to the laboratory for bacterial culture.
ruminants obtained from the samples. Other species isolated were
Bacteriological examination Each nasal swab was removed from the bottle and streaked over
the plates containing blood agar base supplemented with 7% sheep
blood and McConkey agar. The streaking was further spread with
This study showed that a variety of bacterial flora inhabited and
inoculating loop to aid colony isolation. The plates were labeled and
colonized the nasal passageways of apparently healthy West Afri-
incubated aerobically at 37°C for 24-48 h (9). After taking note of
can Dwarf goats. Several authors reported similar bacteria from
cultural growth characteristics, positive cultures were subjected to
pneumonic lungs in goats and sheep (1, 13) with fewer reports in
Gram’s staining properties and cellular morphology observed with a
apparently healthy goats (17). The isolation of S. aureus from
Nasal bacteria isolates from apparently healthy West African Dwarf goats
Bacterial organism Bacterial organisms/ total isolates (%) Streptococcus viridians spp. Flore bactérienne aérobie des cavités nasales de caprins
the nasal passage is consistent with other findings obtained from
caprine (25) and ovine (7) lungs. Robbins et al. (23) report that S. aureus resides in the upper respiratory tract and is involved in dis-
ease processes only when stress conditions prevail. The detection
of E. coli in nasal samples of goats is also consistent with findings
by other authors (20). E. coli, which is known to be usually harm-
less in its normal habitat, can cause pulmonary and urogenital tract
infection (20). This may also be associated with possible fecal con-
tamination due to the sniffing nature of goats, especially those on
heat and during courting before mating. Although Micrococcus spp.
isolated from this study was considered to be non pathogenic (9),
its ubiquity may be primarily due to skin contamination. The isola-
tion of Bacillus spp. in an apparently normal animal was at variance
with other authors’ finding who observed the bacterium in goats’
Augmentin
tracheas and lungs in a disease condition (22), whereas it is sup-
posed to be absent in a healthy animal. The constant isolation of M. haemolytica from the lungs of various animal species either healthy
or having different respiratory syndromes may indicate their pos-
sible role in infectious pneumopathies (16). Hence the isolation of
M. haemolytica and P. multocida in higher proportions (7.9% and
6.4%, respectively) from the nasal cavity of apparently healthy West
African Dwarf goats in this study reflects their possible role in most
common respiratory diseases in small ruminants (8). M. haemolytica was isolated in the nasopharynx and tonsils of
apparently healthy animals, where, interestingly, serotype A2
is most commonly isolated from both sheep and cattle (24). The
organism can be isolated from lambs soon after birth (5) and this
carriage has been shown by Pass and Thompson (18) to fluctuate
over time. The presence of the organism in the nasopharynx of
sheep has been shown to coincide with the occurrence of infec-
tions. In the nasal passages of calves the bacterial flora has been
shown to fluctuate in both species and numbers and, although M. Antibiotic tested haemolytica can dominate the flora, it can also be absent for weeks
at a time (15). There has also been failure to culture consistently M. haemolytica in swabs taken daily from known colonized animals
(18). The mechanisms that M. haemolytica possesses to survive in
Gentamicin
the upper respiratory tract are unknown (24).
Stress factors with or without viral infection have been reported
to suppress the mucociliary clearance mechanism which allows
Ceftriaxone
the proliferation of bacterial commensals in the respiratory tract
(8). They also cause an abrupt shift from commensal to pathogen
especially in M. haemolytica whereserotype 2 shifts to serotype
1, which is known to be pathogenic to animals (12). This shift has
amphenicol
vity of nasal bacteria isolate from apparently health
made M. haemolytica to assume greater prominence in caprine
Chlor ycin
Considering the stress of weather, disease and poor management
conditions to which the animals are constantly subjected, the path-
ogenic role of several bacterial species, and especially M. haemo-lytica, thatinhabit the upper respiratory tract of apparently normal
West African Dwarf goats could be enormous.
Apart from the possible pathogenic role of the normal nasal bac-
terial flora, drug resistance of some the pathogenic bacteria has
become a rampant, proven, serious problem to both animal and
xicillin
ve human health care providers. Aghomo and Ojo (2) reported a
high level of resistance of M. haemolytica to streptomycin while
the organism was found to be susceptible to ampicillin, oxytetra-
cycline, and chloramphenicol. In this study, however, M. haemo-lytica was found to be susceptible to streptomycin, quinolones
(perfloxacin, ciprofloxacin and ofloxacin) and gentamicin, while
it was resistant to tetracycline, augmentin and erythromycin. This
represents a shift of resistance from streptomycin to tetracycline.
This may be associated with the present lesser usage or misuse
Organism
R = resistance; + Susceptible/sensiti of streptomycin for animal diseases unlike tetracycline which is
Aerobic Nasal Bacterial Flora of Goats
commonly used for most animal diseases today. This study shows
14% recovery was obtained (6). However, Ajala et al. (3) obtained
the relationship between misuse or unrestricted use of an antibi-
a 80% recovery rate using Advocin, a newer antibiotic, and a qui-
nolone. Although the survivability of animals affected by the PPR
virus depends on the timing of the commencement of treatment,
The antibiotic sensitivity test on the normal nasal flora of goats also
fluid replacement therapy, and curtailing of secondary bacterial
revealed resistance to cheap and easily accessible antibiotics (tetra-
complications, the use of quinolones, to which most normal nasal
cycline, amoxicillin) which are easily bought off the shelf without
bacteria in this study were susceptible, may be very effective in
appropriate prescription from practicing veterinarians. Apart from
reducing losses during outbreaks as a result of bacterial complica-
ease of accessibility, these drugs have been found to be adulterated
and therefore used at a very low dosage. The drugs to which M. haemolytica was found to be sensitive are expensive and may not be
Therefore, there is a need for practitioners and researchers to be
used frequently. They are also said to be less prone to adulteration.
aware of the aerobic nasal bacterial flora of the West African Dwarf
goats and of their antibiotic sensitivities so as to be informed of the
Since most bacteria complicating PPR in goats are normal com-
appropriate antibiotics to be used in the course of respiratory infec-
mensals of the nasal passages, the results of this study may fur-
ther explain the relationship between the antibiotics employed
and the outcome of treatment management in the reported cases
of PPR virus infection (2, 25). In the studies where penicillin,
Acknowledgments
streptomycin and chloramphenicol were used in the treatment of
We are grateful to the University of Ibadan and to the John D. and
bacterial complications in PPR, less than a 46% recovery rate was
Catherine T. MacArthur foundation grants for staff development,
recorded (26), while with oxytetracycline and chloramphenicol, a
which allowed the first author to complete his PhD program. THOLOGIE INFECTIEUSE PA ■
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Implications for a breeding programme. Small Ruminant Res., 40: 101-108. Flore bactérienne aérobie des cavités nasales de caprins Résumé Resumen Emikpe B.O., Oyero O.G., Akpavie S.O. Isolation et antibio- Emikpe B.O., Oyero O.G., Akpavie S.O. Aislamiento y anti-
gramme de la flore bactérienne aérobie des cavités nasales de
biograma de la flora bacteriana aeróbica nasal en cabras Ena-
chèvres Naines d’Afrique de l’Ouest apparemment saines
Les chèvres jouent un rôle important dans l’économie de l’éle-
Las cabras son importantes en la economía pecuaria por-
vage parce qu’elles sont une source de protéines et de revenus
que son una buena fuente de proteína y de ingresos para los
essentiels pour les pauvres paysans, et parce qu’elles s’adap-
menesterosos rurales y porque se adaptan bien a las condi-
tent à des conditions environnementales difficiles. Les études
ciones adversas del medio ambiente. Los estudios conducidos
sur la flore bactérienne de l’appareil respiratoire des chèvres
sobre la flora bacteriana del tracto respiratorio de cabras se
ont principalement porté sur les lésions pneumotiques ; peu
han enfocado en los pulmones con neumonía con menos estu-
d’études ont été réalisées sur les cavités nasales apparemment
dios en vías nasales aparentemente normales y antibiogramas
normales et peu d’antibiogrammes des organismes isolés ont
de organismos aislados. El presente estudio se llevó a cabo en
été effectués. La présente étude a été menée sur 60 chèvres
60 cabras Enanas Oeste Africanas aparentemente sanas. El fro-
Naines d’Afrique de l’Ouest apparemment saines. Un écou-
tis nasal de cada cabra se analizó usando métodos estándar.
villon nasal a été effectué sur chaque chèvre et analysé avec
La técnica de difusión en disco se empleó para el test de sen-
les méthodes standard. La méthode de diffusion en gélose a
sitividad de antibióticos. Se obtuvieron 328 aislamientos. La
été utilisée pour le test de sensibilité des antibiotiques. Trois
bacteria más frecuentemente aislada fue Streptococcus spp.
cent vingt-huit isolats on été obtenus. Streptococcus spp. a été
mientras que Escherichia coli y Staphylococcus aureus fueron
la bactérie rencontrée le plus fréquemment, suivie d’Escheri-
las segundas especies dominantes. Otras especies fueron tam-
chia coli et de Staphylococcus aureus. D’autres espèces ont
bién aisladas en cantidades relativamente menores. El aisla-
été isolées à des taux relativement plus faibles. La présence
miento de Mannheimia haemolytica y Pasteurella multocida
de Mannheimia haemolytica et de Pasteurella multocida dans
en la cavidad nasal de cabras aparentemente sanas en este
les cavités nasales de chèvres apparemment saines indiquerait
estudio refleja su posible papel en enfermedades respiratorias
leur rôle potentiel dans la plupart des pathologies respiratoi-
comunes encontradas en pequeños rumiantes. La mayoría de
res courantes des petits ruminants. La majorité des bactéries
las bacterias fueron susceptibles a estreptomicina, quinolonas
ont été sensibles à la streptomycine, aux quinolones (perfloxa-
(pefloxacina, ciprofloxacina y eritromicina) y gentamicina,
cine, ciprofloxacine et ofloxacine) et à la gentamicine, alors
mientras que fueron resistentes a tetraciclina, augmentin y
qu’elles ont été résistantes à la tétracycline, à l’augmentin et à
eritromicina. El presente estudio muestra la relación entre el
l’érythromycine. Cette étude a montré la relation entre le mau-
mal uso o uso no restringido de antibióticos y la resistencia
vais usage ou l’abus d’antibiotiques, et la résistance aux médi-
a drogas. En consecuencia, hay una necesidad de informar al
caments. Il est ainsi essentiel d’informer les professionnels de
personal de salud e investigadores sobre los antibióticos apro-
santé et les chercheurs sur les antibiotiques adaptés à utiliser
piados, a usar durante el curso de infecciones respiratorias y
lors d’infections respiratoires et au cours de programmes de
Palabras clave: Caprino – Cabra Enana Africa occidental – Mots-clés : Caprin – Chèvre Naine d’Afrique de l’Ouest – Flore
Flora microbiana – Antibiótico – Resistancia a medicamentos –
microbienne – Antibiotique – Résistance aux médicaments –
Tuberculosis Education Center/TB Elimination Division/Texas Department of Health Phone: (210) 534-8857 ext. 2489, Patient Information Fax: (210) 531-4500 Web Address: www.tdh.state.tx.us/tcid/TB-Education-Ctr.htm 1-800-TEX-LUNG Revised Tuberculosis Treatment Guidelines-2003 For additional information, refer to the printed guidelines. TABLE 2. DRUG REGIMENS FOR CULTURE-POSITIVE PULM
ing algorithms (decision tree and supportvector machine) for HIV-1 genotypic drug-ity is associated with better virological re-Algorithms Need to IncludeHypersusceptibility-resistance interpretation, but neither al-sponses. In addition, hypersusceptibility-persusceptibility. We recently developed alinear statistical model for HIV-1 genotyp-clinical significance has not been firmly To the