Aaa138
Evaluation of Subgingival Bacteria in the Dog and
Susceptibility to Commonly Used Antibiotics
Mirko Radice, DVM; Piera Anna Martino, DBSc, PhD; Alexander M Reiter, Dipl Tzt, Dr med vet
progression from a healthy periodontium to gingivitis andperiodontitis, there is a shift from a gram-positive oriented,
aerobic facultative flora to a predominantly gram-negative,
The aim of the present investigation was to evaluate the
anaerobic flora.22 Periodontopathogens are bacteria that cause
subgingival aerobic and anaerobic flora of 13 dogs with
gingivitis and periodontitis. A catalase-positive form of the gram
periodontal disease and the susceptibility of these bacteria
negative
Porphyromonas gingivalis is considered to be the key
to antibiotics currently approved in Italy for treatment of
periodontopathogen in cats and dogs15,23-25 and is recognized as
P.
canine infections. Of the anaerobic bacteria, Bacteroides
gulae.26 Other canine and feline
Porphyromonas organisms
fragilis was most frequently isolated, followed by
include
P. assacharolytica,
P. cangingivalis,
P. canoris,
P. cansulci,
Peptostreptococcus + Porphyromonas gingivalis and
P. endodontalis,
P. circumdentaria,
P. crevioricanis,
P. salivosa,
P.
Prevotella intermedia. Of the aerobic bacteria, α
-hemolytic
denticanis, and
P. gingivicanis.11,27-30 Additional black-pigmented
Streptococcus was most frequently isolated, often associated
anaerobic bacteria associated with periodontal disease include
with Escherichia coli or Pasteurella multocida. Resistance
Prevotella intermedia23 and
Bacteroides spp.17,18,31,32 Pathogen-
of anaerobic and aerobic bacteria to various antibiotics was
related oral spirochetes also are considered to play an important
generally high. Anaerobic bacteria appeared to be
role in periodontal disease, but cultivation studies in cats and dogs
susceptible to amoxicillin + clavulanic acid, doxycycline,
have only been reported sparsely.8,32-35
and erythromycin; aerobic bacteria appeared to be
Previous studies showed that amoxicillin + clavulanic acid
susceptible to amoxicillin + clavulanic acid, erythromycin,
and clindamycin had high
in vitro susceptibility against anaerobes
gentamycin, and sulfa-trimethoprim. Bacteroides fragilis
and enrofloxacin high
in vitro susceptibility against aerobes from
was resistant to all of the antibiotics tested. The emerging
subgingival plaque samples in cats and dogs.36,37 Although
worldwide problem of bacterial resistance to antibiotics
periodontal disease is caused by bacteria, antibiotic therapy should
resulting from overuse and misuse of antibiotics is
not be the primary treatment strategy.38,39 Unfortunately, there is a
discussed. J Vet Dent 23 (4); #$% - &*, 2006
tendency among veterinarians to use antibiotics as part of themanagement of any animal with periodontal disease or other oralcondition. Resistance of plaque bacteria to antibiotics has clearly
been demonstrated in humans,40-43 and a similar pattern of bacterial
Periodontal disease is an infectious condition of the tooth
resistance development may be present in cats and dogs.
supporting tissues (gingiva, periodontal ligament, alveolar bone,
The aim of the present investigation was to evaluate the
and cementum) and is considered to be the most common disease
subgingival flora (aerobic and anaerobic bacteria) of dogs with
in companion animals. The accumulation of plaque on tooth
periodontal disease and the susceptibility of these bacteria to
surfaces is responsible for the development of gingivitis and
antibiotics currently approved in Italy for treatment of canine
infections. Furthermore, the emerging problem of bacterial
Gingivitis is inflammation of the gingiva and is reversible, if
resistance to antibiotics in human and veterinary medicine is
plaque is removed by home or professional oral hygiene
procedures.2 In addition to toxins and tissue-destructive enzymesproduced by periodontopathogenic bacteria, the host's response to
Materials and Methods
plaque leads to the release of agents from damaged neutrophils
Thirteen client-owned dogs with various degrees of
that can cause injury to the body's own tissues.1 Inflammation
periodontal disease (ranging from gingivitis to periodontitis as
may spread along the periodontal space and ultimately progresses
assessed by means of periodontal probing) were included in this
to periodontitis, which is diagnosed as loss of attachment
study. There were three Yorkshire terriers, one German shepherd,
(gingival recession, resorption of alveolar bone, and formation of
one poodle, and eight mixed-breed dogs. Two dogs received
periodontal pockets). The periapical region of the tooth root may
sporadic oral hygiene at home. A professional scaling and
become affected, leading to retrograde pulpal infection. Thus,
polishing had been performed on all dogs 6-months prior to
endodontic disease can occur as a result of severe periodontal
sample collection. Antibiotics had been given to most of the dogs
disease. Eventually the tooth becomes mobile and is lost due to
in the past for conditions other than periodontal disease. However,
spontaneous exfoliation or professional extraction.1
no attempt was made to assess details of antibiotic history
Gingivitis and periodontitis are referred to as ‘bacterial
because owners were not able to verify names of antibiotics used,
infections', but several hundred bacterial species have been
dates and routes of administration, duration of therapies, etc.
identified to date in normal and diseased mouths of cats and
Dogs had not received antibiotic therapy for at least 2-weeks
dogs.3-21 With maturation of plaque in subgingival areas and
before bacterial sampling. ???
J.VET.DENT. Vol. 23 No. 4 December 2006
219
Sample collection was performed under general anesthesia at
the detachment of microbial cells. Two dilutions (Log10) were
the right maxillary canine tooth (104) and the right maxillary
made for all samples that were plated on Tryptic Soy Agar plates
fourth premolar tooth (108) since both teeth had been excluded
with 5 % sheep bloodb for aerobic bacteria and on Brucella Agarc
from prior periodontal probing. A sterile endodontic paper point
for anaerobic bacteria. The plates were incubated aerobically for
was inserted into the depth of the gingival sulcus or periodontal
24 to 48-hours at 37°C, and anaerobicallyd for 48 to 72-hours at
pocket at buccal aspects of the teeth. The paper point was
removed after a few seconds and placed into tubes containing a
The anaerobic flora was identified by growth on Brucella
transport liquid media (thioglycollate broth)a for growth of
Agare, a medium containing Vit K and haemin. Gram's staining,
aerobic and anaerobic bacteria. The samples were immediately
Schaffer & Fulton's staining for spores, and API System 20Af
transported to the reference laboratory and were vortexed to allow
were used as biochemical reference methods.
Aerobic bacteria were identified by their macroscopic (e.g.,
morphology of colonies, presence of hemolysis) and microscopic
(using Gram-staining) characteristics; moreover, biochemical
Signalment of the 13 dogs enrolled in the study.
tests were performed using macro- or micromethodsg. For theidentification of Streptococcus strains, the presence of hemolysis
(α partial or β total) and the growth on Mitis Salivarius Agarh, a
medium for the isolation and identification of the streptococci of
the oral cavity, were evaluated. For identification of Pasteurella
multocida, the lack of growth on Mac Conkey Agari was
Evaluation of microbial sensitivity/resistance to antibiotics
was performed using the Kirby-Bauer reference method or the
agar disk diffusion test. A bacterial suspension, performed in
saline buffer (0.9 % NaCl), was delivered onto a Mueller-Hinton
plate, and then the disks containing different antibiotic moleculeswere placed on the plate. After incubation at 37°C under aerobic
or anaerobic atmosphere for 24 to 48-hours, the susceptibility of
each microorganism was recorded to the following antibiotics:amikacin, amoxicillin + clavulanic acid, doxycycline,erythromycin, gentamycin, kanamycin, metronidazole (only foranaerobes), and sulfa-trimethoprim.k
Isolation of anaerobic bacteria.
Results of variables of signalment (sex, age, weight, and diet)
Anaerobic bacteria
of the 13 dogs enrolled in the study are reported in Table 1. The
Peptostreptococcus + Porphyromonas
majority of dogs were male (84.6 %), 5 to 10-years of age (53.8
%), < 12.0 kg (61.5 %), and eating a mixed (soft and dry) diet
Bacteroides fragilis
Prevotella intermedia
Of the anaerobic bacteria (Table 2), Bacteroides fragilis was
most frequently isolated from subgingival samples, followed byPeptostreptococcus + Porphyromonas gingivalis, and Prevotella
Anaerobic bacteria
intermedia. Of the aerobic bacteria (Table 3), α-haemolyticStreptococcus was most frequently isolated, often associated with
Peptostreptococcus +
Escherichia coli or Pasteurella multocida.
Porphyromonas gingivalis
Susceptibility of anaerobic and aerobic bacteria to various
antibiotics is shown in Tables 4-8. Resistance of isolated bacteriato tested antibiotics was generally high. Anaerobic bacteriaappeared to be susceptible to amoxicillin + clavulanic acid,doxycycline, and erythromycin, while aerobic bacteria appearedto be susceptible to amoxicillin + clavulanic acid, erythromycin,
gentamycin, and sulfa-trimethoprim. Bacteroides fragilis was
resistant to all of the antibiotics tested.
The aim of the present investigation was to evaluate the
subgingival aerobic and anaerobic bacterial flora of 13 dogs with
J.VET.DENT. Vol. 23 No. 4 December 2006
periodontal disease and the susceptibility of these bacteria to
veterinarians, and diagnostic uncertainty. Several recent studies
antibiotics currently approved in Italy for treatment of canine
showed that pediatricians prescribe antibiotics significantly more
infections. Except for the high prevalence of Bacteroides fragilis,
often, if they perceive parents expect them, and significantly less
the predominant subgingival flora obtained in this study confirms
often, if they feel parents do not expect them.55-58 The best way to
results reported in previous studies.15,17,18,23-25,31,32 Of the anaerobic
combat this situation is to educate patients/patient owners and
bacteria, Bacteroides fragilis was most frequently isolated,
doctors/veterinarians to decrease both demand and over-
followed by Peptostreptococcus + Porphyromonas gingivalis and
prescribing. Unfortunately, there is a tendency to use antibiotics as
Prevotella intermedia. The reason for the unusually high
part of the management of any animal with periodontal disease or
prevalence of Bacteroides fragilis is not clear. Differing results
other oral condition, although there is no apparent justification for
between isolation studies may be due to differences in study
this practice. Similar to dogs in the present study, resistance of
methodology, including sample population utilized and isolation
plaque bacteria to antibiotics has clearly been demonstrated in
techniques applied. Of the aerobic bacteria, α-haemolyticStreptococcus was most frequently isolated, often associated with
Escherichia coli or Pasteurella multocida.
The fast growth rate, high concentration of cells, genetic
Isolation of aerobic bacteria.
processes of mutation and selection, and ability to exchange genesaccount for the extraordinary adaptation and evolution of
bacteria.44 For these reasons bacterial resistance to antibiotics may
α-hemolytic Streptococcus + E. coli
take place very rapidly in evolutionary time. Risk factors
α-hemolytic Streptococcus + P. multocida
responsible for the emergence and spread of resistant bacteria
α-hemolytic Streptococcus + E. coli
include: (1) antibiotic use; (2) reservoirs for resistance; (3)
α-hemolytic Streptococcus + E. coli
medical advances; and (4) societal changes.45 Antibiotics make
conditions favorable for overgrowth of some bacteria, including
α-hemolytic Streptococcus
S. intermedius + P. multocida
those that possess mechanisms of drug resistance. If a resistant
organism is present, antibiotics will create ‘selective pressure'favoring the growth of that organism. A number of studies havedemonstrated conclusively that the development of bacterial
α-hemolytic Streptococcus
resistance to antibiotics is correlated with the level of antibiotic
+ E. coli + P. multocida
use.46-48 Antibiotic resistance of nosocomial pathogens in hospitals,
α-hemolytic Streptococcus
+ E. coli + S. intermedius
nursing homes, day-care centers, and animal facilities is increasedby the transfer of individuals already colonized by resistant
+ S. intermedius
organisms from one location to another. Progress in the treatment
of many diseases has led to an increased life span of humans andanimals. Consequently, with advanced age, chronic disease orimmunosuppression, individuals can be more susceptible tobacterial infections, resulting in greater use of antibiotics.
Worldwide spread of bacterial resistance to antibiotics hasoccurred due to the increased mobility of today's society.45
Pet animal numbers have substantially increased in modern
α-hemolytic Streptococcus + P. multocida
society, and attention is increasingly devoted to pet welfare.
Antibiotics are frequently used in small animal practice, withheavy use of broad-spectrum agents such as amoxicillin +clavulanic acid, cephalosporins, and fluoroquinolones. The
practice of antibiotic overuse and misuse in cats and dogs hascontributed to the development of Staphylococcus spp.,
Antibiotic activity versus Prevotella intermedia.
Escherichia coli and various other bacteria that are resistant toantibiotics.49-53 The role of pets in the dissemination of bacterial
resistance to antibiotics has been given relatively little attention
when compared with that of food animals, and a marked contrast
is evident between the current policies on antibiotic usage in
companion and food animals. However, the possible transfer of
resistant bacteria from cats and dogs to humans has recently been
acknowledged as a potential threat to public health.44,54
One of the biggest problems is inappropriate prescribing of
antibiotics. There are many reasons for this, including demand
from patients/patient owners, time pressure on physicians/
J.VET.DENT. Vol. 23 No. 4 December 2006 221
Antibiotic activity versus Bacteroides fragilis.
Antibiotic activity versus Porphyromonas gingivalis +Peptostreptococcus.
Antibiotic activity versus α-haemolytic Streptococcus + E.
Antibiotic activity versus α-haemolytic Streptococcus + P.
humans.40-43 It is therefore imperative to review periodontaltreatment strategies and determine whether systemic antibiotics
have a role to play in the management of periodontal disease.
If accumulation of plaque is prevented, periodontal disease
American Veterinary Dental College (AVDC) Position
does not develop.2 Although this condition is caused by bacteria,
Statement on the Use of Antibiotics in Veterinary Dentistry.
antibiotic therapy is not considered the primary treatmentstrategy.38,39 Instead, treatment of periodontal disease should be
The AVDC endorses the use of systemic antibiotics in
directed at mechanical removal or reduction of plaque and
veterinary dentistry for treatment of some infectious
calculus accumulation, suppression of the tissue-destructive
conditions of the oral cavity. Although culture andsusceptibility testing is rarely performed on individual
effects of the inflammatory response, surgical management of
patients that have an infection extending from/to the oral
periodontal pockets, extraction of more severely affected teeth,
cavity, the selection of an appropriate antibiotic should be
and thorough debridement of extraction sites.1 A controlled-
based on published data regarding susceptibility testing
release local antibiotic delivery system, reaching
of the spectra of known oral pathogens. Patients that are
periodontopathogens deep within periodontal pockets, has been
scheduled for an oral procedure may benefit from pre-treatment with an appropriate antibiotic to improve the
described in dogs.59 Professional supra- and subgingival scaling,
health of infected oral tissues. Bacteremia is a recognized
followed by daily tooth brushing, is the ‘gold standard' for
sequela to dental scaling and other oral procedures.
prevention of periodontal disease.1 Home oral hygiene may be
Healthy animals are able to overcome this bacteremia
enhanced by offering products that support dietary abrasion or
without the use of systemic antibiotics. However, use of asystemically administered antibiotic is recommended to
chemically suppress plaque and calculus accumulation.60
reduce bacteremia for animals that are immune
Bacteremia secondary to periodontal disease occurs daily in
compromised, have underlying systemic disease (such
patients with periodontal disease, and it is normally rapidly
as clinically-evident cardiac, hepatic, and renal diseases)
cleared by the reticulo-endothelial system in the healthy patient.61
and/or when severe oral infection is present. Antibiotics
Therefore, for the great majority of otherwise healthy cats and
should never be considered a monotherapy for treatmentof oral infections, and should not be used as preventive
dogs presenting with periodontal disease and other oral
management of oral conditions. Adopted by the Board of
conditions, systemic antibiotics are not indicated.1,39 Bacteremia
Directors, April 2005
can be prevented or reduced in severity by rinsing the oral cavity
J.VET.DENT. Vol. 23 No. 4 December 2006
with dilute chlorhexidine gluconate (0.12 %) prior to
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commencing the oral procedure. Perioperative systemicantibiotics are indicated in: (1) debilitated and
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Source: http://www.dentalvet.it/wp-content/uploads/2016/03/Evaluation-of-Subgingival-Bacteria-in-the-Dog-pubblicato3.pdf
ORIGINAL RESEARCH ARTICLE Influence of Antihypertensive Therapy on Cerebral Perfusion inPatients with Metabolic Syndrome: Relationship with CognitiveFunction and 24-h Arterial Blood Pressure Monitoring Nataliya Y. Efimova,1,2 Vladimir I. Chernov,1,2 Irina Y. Efimova1 & Yuri B. Lishmanov1,2 1 Federal State Budgetary Scientific Institution, Research Institute for Cardiology, Tomsk, Russia2 National Research Tomsk Polytechnic University, Tomsk, Russia
HIGHLIGHTS OF PRESCRIBING INFORMATION Suicidal Behavior and Ideation (5.1) These highlights do not include all the information needed to use Patients should be advised that VIMPAT may cause dizziness VIMPAT® safely and effectively. See full prescribing information and ataxia. (5.2) for VIMPAT.