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Introduction
International Journal of Poultry Science 7 (11): 1058-1064, 2008ISSN 1682-8356 Asian Network for Scientific Information, 2008
Susceptibilities of Mycoplasma gallisepticum and Mycoplasma synoviea Isolates to Antimicrobial Agents in vitro
Razi Vaccine and Serum Research Institute, Shiraz, Iran
Poultry Research Centre, Veterinary School, Shiraz University, Shiraz, Iran
Agriculture School, Shiraz University, Shiraz, Iran
Mycoplasma Reference Lab. of Razi Vaccine and Serum Research Institute, Karaj, Iran
Abstract: The in vitro activities of aivlosin tylosin tiamulin, erythromycin, oxytetracycline, spectinomycin, lincomycin, lincomycin-spectinomycin (1:2) and enrofloxacin were determined against twenty-eight isolates of Mycoplasma gallisepticum and 4 isolates of Mycoplasma synoviae using microbroth dilution method. The isolates showed various susceptibilities to antimicrobial agents. Aivlosin, lincomycin-spectinomycin (1:2), tylosin, tiamulin, enrofloxacin and also lincomycin were commonly more effective against these field isolates. However, aivlosin was the most effective drugs against Mycoplasma gallisepticum and Mycoplasma synoviae as its MIC was the lowest of all. Oxytetracycline, erythromycin and spectinomycin were not effective against all isolates, as their MICs for some of isolates were extremely high. The MIC of erythromycin and oxytetracycline were distributed across a broad range. Resistant strains to two mentioned antibiotics were obtained from the field. Key words:Mycoplasma sp., antimicrobial agents, poultry INTRODUCTION
vertical transmissions. Here the most of veterinarians
Mycoplasma gallisepticum and Mycoplasma synoviae
prescript the consumption of antibiotics on the basis of
has long been recognized as common respiratory
clinical findings and their experiences in order to
pathogens especially in chickens causing lots of
treatment affected flocks and improve egg production
commercial losses in poultry industries. Mycoplasma
rate. It is logical that for a successful and aimed
gallisepticum infection commonly induces chronic
mycoplasma infection treatment, it is necessary to have
respiratory disease in chickens (Ley, 2003).
regular antibiogram tests of Mycoplasma gallisepticum
The clinical signs include nasal discharge, sneezing,
and or Mycoplasma synoviae in the field for monitoring
coughing, tracheal rales and mild conjunctivitis.
susceptibility of Mycoplasma prevalent in the farms. Mycoplasma synoviae most frequently occurs as asubclinical upper respiratory infection but may result in
Antibiotics: Basically Mycoplasma gallisepticum and
airsacculitis and synovitis in chickens and turkeys
Mycoplasma synoviae have shown sensitivity in vitro and
in vivo to several antimicrobials including macrolides
Establishing of the Mycoplasma gallisepticum and
(Jordan and Horrocks, 1996) tetracyclines and
Mycoplasma synoviae-clean status of breeder flocks and
quinolones (Bébéar et al., 1999; Wu et al., 2000), but
maintaining that status of can be accomplished by
they are resistant to penicillins or other antibiotic
participation in control programs (Kleven, 2003; Ley,
2003). Like other countries in Iran chicken primary and
On the other hand many antimicrobial agents, such as
multiplier breeders and hatcheries generally have
oxytetracycline, amino glycosides lincosamides,
adopted various mycoplasma control programs. In spite
fluoroquinolones and tiamulin have been shown to
of these preventive programs, a lot of broiler and layer
possess different degrees of in vitro activity against
flocks will be affected by those organisms during their
various veterinary mycoplasmas (Bradbury et al., 1994;
production periods. However , line and parent flocks are
Hannan et al., 1997a). However, increasing resistance
mycoplasma free, but a large number of breeder flocks
of mycoplasmas against tetracyclines (Hannan et al.
also involve Mycoplasma gallisepticum and or
1997a) macrolides (Bradbury et al., 1994; Hannan et al.,
Mycoplasma synoviae infections. However, it is not
1997a; Gautier-Bouchardon et al., 2002) and quinolones
widely allowed the use of lived mycoplasma vaccines in
(Bébéar et al., 1999; Wu et al., 2000) has been reported
the country, control of mycoplasma infections by
vaccination is limited. Control of these infections by chemotherapy is
MATERIALS AND METHODS
sometimes necessary in complement of biosecurity
On this way we carried out a 2 stage research, at first
measures to minimize economic losses and lateral and
stage isolation and molecular identification of
Int, J. Poult. Sci., 7 (11): 1058-1064, 2008Mycoplasma gallisepticum and Mycoplasma synoviae
broader spectrum of activity than the macrolides, being
were done. Isolation was carried out by enriched
active also against a lot of gram-negative bacteria, which
mycoplasma broth and agar. One hundred mycoplasma
very often complicate Mycoplasma gallisepticum and
isolates were identified genetically by PCR and RFLP
Mycoplasma synoviea infections (Ghaleh et al., 2005).
(Garcia et al., 1995; Ghaleh et al., 2005). All of isolates
At first 0.2 mL was chosen as final volume for each well
gathered in Fars province of Iran and related tests
containing 0.05 mL antibiotic solution, 0.05 mL fresh
carried out in Shiraz Veterinary Medicin School, Shiraz,
broth media solution (and then serially diluted) finally 0.1
mL fresh broth culture of each isolates inoculated toeach well (which they made 0.2 mL of test volume). Test organisms: In the second stage the antibiogram
However fresh broth culture of each isolates used as
test were carried out on 28 field isolates of Mycoplasma
primary inoculums, the number of viable mycoplasmas
gallisepticum and 4 field isolates Mycoplasma synoviae
inoculated in each test of sensitivity to the antibiotics
by common antibiotics. The antibiogram test was
was determined by micro-broth dilution using
designed on the basis of micro dilution method in broth
mycoplasma enrichment broth with phenol-red indicator.
medium (Whithear et al., 1983; Hannan et al., 1997b).
Although, considerable data has been obtained on the
was needed for the test proper which is added to each
in vitro antibiotic susceptibility of
well without any serial dilution. The change of color in
gallisepticum and Mycoplasma synoviae and other avian
the broth medium from red to yellow following
mycoplasmas abroad but no information is available
inoculation of isolates and incubation at 37°C for 14
about the susceptibility of these organisms in Iran. All
days showed growth of the isolates. A micro-broth
isolates were between the 20th and 30th medium
dilution procedure which is described by Whithear et al.
(1983) used in this research except we count thecolonies of 24 fresh broth cultures of isolates before
Antibiotics: Antibiotics used to determine the sensitivity
starting the sensitivity tests. The test was carried out in
of Mycoplasma gallisepticum and Mycoplasma synoviae
96-well, U bottom micro titration disposable plates with
isolates, were Aivlosin tartrate (E.Co.A.H.) Tylosin tartrate
cover under sterile conditions using multichannel
(Sigma), Tiamulin base (ACS Chemicals), Erythromycin
micropipette (Ependorffe Co., Germany). At least six and
base (Merck), Oxytetracycline dihydrate (Sigma),
sometimes 10-14 replicate doubling dilutions were
Spectinomycin dihydrochloride (Sigma), Lincomycin
made in experiments to evaluate test variables.
hydrochloride (Sigma), Lincomycin-Spectinomycin (1:2),
According to Table 1, 150 µL of mycoplasma enrichment
Enrofloxacin base (ACS Chemicals). Erythromycin base
broth (pH 7.4) were added to each well containing final
were dissolved in 7% ethanol. Each antibiotic solution
dilution of antibiotic, containing desired density of
was passed through a 450 nm pore size membrane
isolates was inoculated in to each well. Positive and
filter (Millipore, USA) and then dispensed aseptically.
negative culture controls containing mycoplasma
FOR every antibiogram test the solutions prepared
enrichment broth plus different isolates without
under sterile conditions and in WFI water as stock
antibiotics and mycoplasma broth plus different isolates
solutions. The stock solutions were diluted in
and antibiotics and zero culture controls included only
mycoplasma broth to dilution two times that required
mycoplasma enrichment broth (without any isolates and
and were dispensed in 1 mL aliquots and then frozen at
antibiotics), were included in all test plates. Plates were
covered with their covers to prevent the evaporation ofthe broth medium during incubation period at 37°C. As
Media: The media used were the mycoplasma broth and
it mentioned above the standardized test microbroth
or mycoplasma agar. Mycoplasma broth medium was
dilution procedure which is described by Whithear et al.
formulated and described by Frey et al. (1968). Added
(1983) was applied for Mycoplasma gallisepticum and
glucose (dextrose) to broth medium mycoplasma
Mycoplasma synoviae isolates. The standardized test
fermentative growth was enhanced and provided an
was used to determine the sensitivity of 28 strains of
indicator of growth when glucose fermentation produced
Mycoplasma gallisepticum, four strains of Mycoplasma
acids in the media containing phenol red. The color of
synoviae to the antibiotics. Mycoplasma broth prepared
medium gradually changed from red to orange or yellow
by the described formula. Erythromycin base were
if the growing fermentative species of mycoplasmas
dissolved in 7% ethanol. Each antibiotic solution was
passed through a 450 nm membrane filter (Millipore,
Sometimes the effectiveness of various antibiotics
USA) and then dispensed aseptically. For every
towards Mycoplasma gallisepticum and Mycoplasma
antibiogram test antibiotic solutions were prepared
synoviea in the field is not very different but some
freshly and immediately used. Solutions were diluted in
antibiotics show better advantages. Enrofloxacin still
mycoplasma broth in two fold dilution started in 100
retains the acceptable advantage because it has
µg/mL for first time but for second and third time the
Int, J. Poult. Sci., 7 (11): 1058-1064, 2008
Table 1: In vitro activities of, oxytetracycline, aivlosin, tylosin, tiamulin, erythromycin, spectinomycin, lincomycin, L/S and enrofloxacin against
*Isolate numbers 23, 24, 31 and 32 are Mycoplasma synoviae (MS) see Table 2
narrower rage of titration was required started in 25, 6.25
and 0.195 µg/mL. The ranges of concentrations of active
The in vitro activities of lincomycin-spectinomycin
compounds used in the final tests to survey the
combination (1:2), erythromycin, oxytetracycline,
sensitivity of avian mycoplasmas to the antibiotics were
lincomycin, spectinomycin, enrofloxacin, tylosin, aivlosin
as follows: Tiamulin 25 µg/mL to 1.56 µg/mL,
and tiamulin against the 28 isolates of Mycoplasma
spectinomycin, oxytetracycline, erythromycin, lincomycin,
gallisepticum and 4 isolates of Mycoplasma synoviae,
tylosin and enrofloxacin from 6.25 µg/mL to 0.195 µg/mL
as determined by the micro-broth technique, are shown
to 0.012 µg/mL (Hannan et al., 1997a,b). All antibiotic
in Table 1. Of the eight antimicrobials, oxytetracycline
solution pH were adjusted to the pH of the mycoplsma
had the highest and aivlosin had the lowest MIC values.
The results for Mycoplasma synoviae are given in Table2 and again demonstrate the greater sensitivity of the
Antibiogram tests were carried out in 96 well sterilized
isolates to aivlosin and lincomycin-spectinomycin than
micro plate U bottoms. When growth occurred in the
to oxytetracycline. Of the other antimicrobials
well, a definite change was observed of the phenol red
enrofloxacin, tylosin erythromycin, tiamulin, spectino-
to yellow (usually after 140 h of incubation) and the
mycin and lincomycin showed the lowest MICs with
minimum inhibitory concentration (MIC) was determined.
The MIC was the lowest concentration of antibiotic
Against 4 isolates of MS by the micro-broth method
inhibiting growth (no color change of phenol red).
aivlosin showed the best in vitro activity giving MICs of
When the MIC was determined, approximately 0.01 mL
0.042 µg/mL for most isolates. Also, lincomycin-
of broth from each tube was placed on an agar plate.
spectinomycin (1:2) combination had MICs of
The plates were incubated at 37ºC in a moist container
0.0485/0.097 µg/mL for most isolates. Tiamulin and
and examined microscopically after 5 days for
erythromycin had similar MICs of 0.39-0.78 µg/mL for the
mycoplasma colonies (Gautier-Bouchardon et al., 2002,
isolates. Spectinomycin and lincomycin had MICs
Bradbury et al., 1994). All MIC values were expressed in
ranging from 0.78-1.56 µg/mL and oxytetracycline had
µg/mL of each active compound. Also all sensitivity tests
repeated second times to determine the accuracy of
The combination of lincomycin-spectinomycin (1:2)
appeared to have a synergistic effect for all isolates. Int, J. Poult. Sci., 7 (11): 1058-1064, 2008In vitro activities of oxytetracycline, aivlosin, tylosin, tiamulin, erythromycin, spectinomycin, lincomycin, lincomycin-spectinomycin andenrofloxacin against Mycoplasma synoviae isolates
According to the results all MS strains are resistant to
Review of the published literatures describing sensitivity
oxytetracycline and sensitive to aivlosin, tylosin, tiamulin,
testing of mycoplasmas reveals that there is
lincomycin, lincomycin - spectinomycin combination
considerable various workers on this title. There
(1:2) and enrofloxacin. These strains were sensitive to
spectinomycin except isolate No.32 and semi-sensitive
mycoplasmologists about standard procedures for
or resistant to erythromycin except isolate No. 23.
testing of mycoplasmas for sensitivity to antibiotics, forexample, Lin had been used agar dilution method to
DISCUSSION
determine susceptibility of avian mycoplasmas to
The purpose of the study was to determine the in vitro
antibiotics (Lin, 1987) Boughton also demonstrated
susceptibilities of Mycoplasma gellisepticum and
tylosin susceptibility in 10 isolates of MycoplasmaMycoplasma synoviae, which isolated from broiler
gallisepticum tested by disc diffusion techniques
chicken farms, Fars, Iran. in vitro susceptibility testing of
mycoplasmas presents several problems that make
In this research the MICs were determined by a glucose
standardization of methods difficult. Because no single
metabolic inhibition method. Therefore, a micro-broth
medium is suitable for in vitro testing of all species, we
dilution procedure was chosen as the basic test design,
had to use a different medium for each of the two
because it was relatively simple and quick to perform
species. It was recommended to add NAD to mediums
and required only small volumes of media and other
for cultivating Mycoplasma synoviea. A pH indicator such
necessary materials such as different antibiotics. The
as phenol red visualizes the pH shift due to biochemical
test was also easy to read, since inhibition of
activities of multiplying mycoplasmas. One of the major
mycoplasma growth by antibiotics could be deduced by
ways for Mycoplasma gallisepticum and Mycoplasma
the inability of the organism to ferment glucose and thus
synoviae control is antibiotic therapy of affected flocks
to produce a color change in a phenol red indicator in
and infected hatching eggs (Alls et al., 1963). In Iran
the medium from red to orange-yellow and generally
antibiotic administration has been accepted and
more acceptable between mycoplasma scientists. As
different antibiotics are available and administered
mentioned before, replicate doubling dilutions of
against mycoplasma infection for several decades. This
antibiotics were made in 150 microliter volumes of
is important to know that administration of antibiotics
mycoplasma broth in sterile 96-well U-bottomed micro
despite of dosage and duration of administration could
titration plates (Hirose et al., 2003; Gautier-Bouchardon
not eradicate these organisms from the infected flocks. et al., 2002; Hannan et al., 1997a; Whithear et al., 1983)
But antibiotics are able to reduce the severity of clinical
using 8-channel micropipette (Ependorff, Germany). The
signs and lesions, decrease egg production losses and
test was read when the phenol red indicator in the
other economic losses such as downgrading
medium of a culture control (without antibiotic) had
carcasses. Proper antibiotic therapy could significantly
changed from deep red (pH 7.8) to orange-yellow (pH 7).
reduce population of Mycoplasma gallisepticum and
It is believed that the number of viable mycoplasmas
Mycoplasma synoviea in the poultry respiratory tract.
inoculated in each test of sensitivity to antibiotic was very
Mycoplasma gallisepticum and Mycoplasma synoviea
important to reliability of the test. In this way to determine
have shown sensitivity in vitro to several antimicrobials
the concentration of each isolates in the mycoplasma
broth and for making more accuracy the colony counting
fluoro-quinolones and others, but also are resistant to
procedure was done (Hirose et al., 2003; Zanella et al.,
other antibiotics such as cephalosporins and penicillins
1998). Some of other scientists preferred to determine
which act by inhibiting cell wall biosynthesis.
the number of viable organisms by color changing unit
Mycoplasma gallisepticum may develop resistance and
(CCU) in the original culture by consulting the tables
even demonstrate cross-resistance, to commonly used
which were published by Meynell and Meynell (1970),
antibiotics (Bradbury et al., 1994; Zanella et al., 1998).
(Hannan et al., 1997a,b; Whithear et al., 1983).
There appears to be no general agreement among
There is a wide range of bacteria and mycoplasma
mycoplasmologist about standard procedures for
susceptibility to antibiotics in different area and
testing of mycoplasmas for sensitivity to antibiotics.
countries. On the other hand another problem with
Int, J. Poult. Sci., 7 (11): 1058-1064, 2008
antibiogram test of mycoplasma is the different growth
less sensitive to tylosin. However, in our research the
Mycoplasma gallisepticum and Mycoplasma synoviea
According to the results of our research, 28 isolates of
isolates were more sensitive to tylosin than
Mycoplasma gallisepticum and four isolates of
Mycoplasma synoviea showed various susceptibilities
Aivlosin, lincomycin-spectinomycin (1:2), tylosin,
to the nine different antimicrobial agents which used in
tiamulin, enrofloxacin and also lincomycin were
commonly very effective in the field isolates. These
The breakpoints of MICs of each antimicrobial agent
findings are similar to the reports of Hannan (2000) and
group: oxytetracycline, macrolides (aivlosin, tylosin and
Zanella (1998). Although, macrolides and oxytetracycline
erythromycin), aminoglycoside (including spectino-
were considered to be effective against mycoplasmas,
mycin), lincosamides (lincomycin), fluoroquinolone
these drugs (erythromycin and oxytetracycline) did not
(enrofloxacin) and tiamulin were 1.0, 0.5-2.0, 2.0-4.0 and
prove to be effective against Mycoplasma synoviea
1-2 µg/mL, 0.5-2.0 and 0.1-0.05 µg/mL, respectively
isolates. These MICs are higher than breakpoints of
(Hirose et al., 2003). When the MIC of the oxytetracycline
macrolides (0.5-0.2 µg/mL) or oxytetracycline (1 µg/mL).
was =1.0 µg/mL the isolate was considered susceptible,
According to the commercial costs of used antibiotics
when the MIC was >1.0 µg/mL, the isolate was
against Mycoplasma gallisepticum and Mycoplasmasynoviea tetracyclines such as oxytetracyline is not
In contrast to Mycoplasma gallisepticum isolates, all of
expensive and have been previously used in large
Mycoplasma synoviea isolates have been shown
scales to control and treat suspected broiler flocks to
resistance to oxytetracycline and almost erythromycin.
mycoplasmaand E. coli infections and now observed
This finding is in complete agreement with research
that mycoplasma showed highly resistance to these
results of Bradbury et al. (1994) and Whithear et al.
kinds of antibiotics. On the other hand, aivlosin are more
(1983). Also, they used micro-broth dilution procedure
expensive then its use has limited. Also, aivlosin hasbeen more recently introduced to producer. Also, our
for MIC detection in their research. The results also in
field experiences support these findings.
agreement with reports of Kleven and Anderson (1971)
However, aivlosin was one of the most effective drugs
and also Bradbury (1994) in the case of MICs of
against Mycoplasma gallisepticum and Mycoplasma
lincomycin-spectinomycin in comparison with
synoviea as its MIC was the lowest of all oxytetracycline,
oxytetracycline. Results of the present study agreed with
erythromycin and spectinomycin were not effective
those of Tanner and Wu (1992) who used broth micro-
against all isolates, as its MIC for some of isolates was
dilution test and showed Mycoplasma gallisepticum
extremely high. The MIC of erythromycin and
isolates were susceptible to lincomycin-spectinomycin
oxytetracycline were distributed across a broad range.
and in the case of erythromycin and tylosin who found
Resistant isolates to two mentioned antibiotics were
the susceptibility varied among their isolates that is
similar to our findings. But they reported their isolates
Results showed that the sensitivity of isolates to
were sensitive to oxytetracycline. On the other hand
spectinomycin and licomycin lonely was less than their
Bradbury (1994) reported that tylosin had the highest
sensitivity to lincomycin-spectinomycin in 1:2
activity followed closely by lincomycin, oxytetracycline,
spectinomycin. Erythromycin was less effective, but like
Sometimes the efficacy of various antibiotics towards
our findings lincomycin-spectinomycin had good activity
Mycoplasma gallisepticum and Mycoplasma synoviea in
against Mycoplasma gallisepticum.
the field is equal but some antibiotics show better
According to our research aivlosin had greater activity
advantages. Enrofloxacin still retains the acceptable
during 9 antibiotics used with a range 0.012-0.042
advantage because it has broader spectrum of activity
µg/mL. The reason may be aivlosin is a newer antibiotic
than the macrolides, being active also against a lot of
which is recently used in Iran against Mycoplasma
gram-negative bacteria, which very often complicate
gallisepticum and Mycoplasma synoviea. However,
Mycoplasma gallisepticum and Mycoplasma synoviea
according to our knowledge the use of aivlosin in poultry
is relatively limited in Europe and not at all in the USA.
In some cases when an appropriate antibiotic, on the
Comparison of MIC values with tylosin and tilmicosin
basis of antibiogram test, administrates the results may
would be interesting. Zanella et al. (1998) investigating
be out of our prediction and treatment results are not
development antibiotics resistance in Mycoplasma
satisfy. It seems that some of avian respiratory viruses
gallisepticum, also reported that cross-sensitivity tests
such as infectious bronchitis virus newcastle disease
using strains with induced resistance to the different
virus and pneumoviruses and also E. coli may interfere
antibiotics demonstrated that those which were resistant
with Mycoplasma gallisepticum and Mycoplasma
to tylosin were also resistant to other macrolides, where
synoviea antibacterial treatments (Kleven, 1998a, b).
as strains made resistant to erythromycin appeared only
The treatment of these cases may be difficult because of
Int, J. Poult. Sci., 7 (11): 1058-1064, 2008
some viral coinfections such as infectious bronchitis,
Conclusion: Whichever method of control is chosen, the
avian influenza or infectious bursal disease and then
need for effective antibiotics is apparent. According to
infection may remain for a long period. This finding is in
our findings drug resistance must be taken into
agreement with other reports (Kleven and Anderson,
consideration in mycoplasma and this fact stresses the
1971; Bradbury et al., 1994). Therefore, some of
desirability for a choice between several antibiotics. The
countries prefer to eradicate mycoplasma infections
in vitro sensitivity of 28 different isolates of Mycoplasma
from their flocks by test and slaughter strategy.
gallisepticum and 4 isolate Mycoplasma synoviea havebeen tested using 9 different antibiotics includingaivlosin and lincomycin-spectinomycin (1:2). These 2
Specimens in Iran: Ever since the establishment of the
antibiotics showed the highest effect followed by tylosin,
sensitivity to broad spectrum antibiotics (such as
enrofloxacin, tiamulin and lincomycin.
tetracyclines, macrolides and etc.) of Mycoplasmagallisepticum and Mycoplasma synoviea these and
ACKNOWLEDGEMENT
more recently even other antibiotics like tylosin and
I wish to thank Dr. Rajaeeyan for his cooperation and
erythromycin have been widely used in both preventive
preparing the different antibiotics at Pharmacology Dept.
and curative treatment of mycoplasma infection in
of Shiraz Veterinary School. This research was
poultry. This is true even when programs for the creation
financially supported by Shiraz University.
of mycoplasma-free flocks are applied in mycoplasmacontrol and antibiotic treatment also is an important tool
REFERENCES
in the efforts to control mycoplasmosis by breaking the
Alls, A.A., W.J. Benton, W.C. Krauss and M.S. Cover,
disease transmission chain via the hatching egg which
1963. The mechanics of treating hatching eggs for
is unanimously claimed to be the most important route
disease prevention. Avian Dis., 7: 89-97.
Bébéar, C.M., J. Renaudin, A. Charron, H. Renaudin, B.
Unfortunately despite of our review article, we could not
de Barbeyrac, T. Schaeverbeke and C. Bébéar,
find any report about mycoplasma antibiogram studies
1999. Mutations in the gyrA, parC and par E genes
especially in poultry in the country. Now in Iran, the
associated with fluoroquinolones resistance in
veterinary practitioners usually come to antibiotic
clinical isolates of Mycoplasma hominis. Antimicrob.
treatment of mycoplasmal diseases of poultry on the
Boughton, E., 1982. Antibiotic sensitivities of
basis of clinical signs of the affected poultries without
mycoplasmas. Vet. Rec., 111: 541-542.
any paraclinical findings and or antibiogram test before
Bradbury, J.M., C.A. Yavari and C.J. Giles, 1994. In vitro
it. The results of this kind of blind antibiotic therapy are
evaluation of various antimicrobial against
very harmful for human, poultry and life environment and
The potential for mutation and for genetic exchange
comparison with a commercially prepared system.
between all types of bacteria including mycoplasmas,
combined with the short bacterial generation time, is of
Frey, M.L., R.P. Hanson and D.P. Anderson, 1968. A
major importance in limiting the use of antimicrobial
medium for the isolation of avian mycoplasmas.
drugs in controlling infection in poultry. Sometimes, the
blind use of antimicrobial drugs may does not induce
Garcia, M, M.W. Jackwood, S. Levisohn and S.H. Kleven,
resistance in bacteria but rather eliminates the
1995. Detection of Mycoplasma gallisepticum, M.
susceptible bacteria and leaves the resistant bacteria
synoviae and M. iowae by multi-species polymerase
chain reaction and restriction fragment length
These above findings should be available to poultry
polymorphism. Avian Dis., 39: 606-616.
disease specialists and veterinarians, as they consider
Gautier-Bouchardon, A.V., A.K. Reinhardt, M. Kobisch
therapeutic approaches to CRD and other poultry
and I. Kempf, 2002. In vitro development ofresistance to enrofloxacin, tylosin, tiamulin and
mycoplasmal diseases. Therefore, it is necessary to
have a regular survey on poultry pathogenic
Mycoplasma iowae and Mycoplasma synoviea. Vet.
mycoplasmas and to monitor antimicrobial susceptibility
patterns in order to ensure that effective chemotherapy
Ghaleh, G.B.N., K. Asasi, A.R. Afsharifar and S.A.
is being used to treat mycoplasmal infections.
Pourbakhsh, 2005. Isolation and detection of
However, mycoplasmal isolation, identification and
Mycoplasma gallisepticum by polymerase chain
antimicrobial susceptibility tests are essential and
helpful for choosing an appropriate chemotherapy
polymorphism. Iranian J. Vet. Res. Univ. Shiraz, 6:
Int, J. Poult. Sci., 7 (11): 1058-1064, 2008
Hannan, P.T.C., G.D. Windsor, A. de Jong, N. Schmeer
Kleven, S.H. and D.P. Anderson, 1971. In vitro activity of
various antibiotics against Mycoplasma synoviae.
susceptibilities of various animal-pathogenes
mycoplasmas to fluoroquinolones. Antimicrob.
Ley, D.H., 2003. Mycoplasma gallisepticum Infection. In:
Saif, Y.M., H.J. Barnes, J.R. Glisson, A.M. Fadly, L.R.
Hannan, P.T.C., H.M. Windsor and P.H. Ripey, 1997b. In
McDougald and D.E. Swayne (Eds.). Diseases of
vitro susceptibilities of recent field isolates of
Poultry. Eleventh ed. Ames. Iowa, Iowa State Press,
Mycoplasma hyopneumoniae and Mycoplasma
A Blackwell Publishing Company, USA., pp: 722-
hyosynoviae to valnemulin (Econor), tiamulin and
enrofloxacin and the in vitro development of
Lin, M.Y., 1987. In vitro comparison of the activity of
resistance to certain antimicrobial agents in
various antibiotics and drugs against new Taiwan
Mycoplasma hyopneumoniae. Res. Vet. Sci., 63:
isolates and standard strain of avian Mycoplasma.
Hirose, K., H. Kobayashi, N. Ito, Y. Kawasaki, M. Zako, H.
Meynell, G.G. and E.W. Meynell, 1970. Theory and
mycoplasmas from nasal swabs of calves affected
practice in experimental bacteriology. 2nd Edn.
with respiratory diseases and antimicrobial
Cambridge University Press. Cambridge.
susceptibility of their isolates. J. Vet. Med. Series B.,
Tanner, A.C. and C.C. Wu, 1992. Adaptation of
Sensititre broth microdilution technique to
Jordan, F.T.W. and B.K. Horrocks, 1996. The minimum
antimicrobial susceptibility testing of Mycoplasma
inhibitory concentration of tilmicosin and tylosin for
gallisepticum. Avian Dis., 36: 714-717.
Whithear, K.G., D.D. Bowtell, E. Ghiocas and K.L. synoviae and a comparison of their efficacy in the
Hughes, 1983. Evaluation and use of a Micro-broth
control of Mycoplasma gallisepticum in broiler
dilution procedure for testing sensitivity. Avian Dis.,
Kleven, S.H., 2003. Mycoplasma synoviae Infection. In:
Wu, C.C., T.R. Shryock, T.L. Lin and M. Faderan, 2000.
Saif, Y.M., H.J. Barnes, J.R. Glisson, A. M. Fadly, L.R.
McDougald and D.E. Swayne (Eds.), pp: 756-766.
hyorhinis. Vet. Microbiol., 76: 25-30.
Kleven, S.H., 1998a. Mycoplasmas in the etiology of
Zanella, A., P.A. Martino, A. Pratelli and M. Stonfer, 1998.
multifactorial respiratory disease. Poult. Sci., 77:
Development of antibiotic resistance in Mycoplasmagallisepticumin vitro. Avian Pathol., 27: 591-596.
Kleven, S.H., 1998b. Mycoplasmosis. In: Swayne, D.E.,
J.R. Glisson, M.W. Jackwood, J.E. Pearson andW.M. Reed (Eds.). A laboratory manual for theisolation of avian pathogens. 4th Edn. AmericanAssociation of Avian Pathologists: Kennett Square,PA, pp: 74-80.
China’s Hisun Files IND For Cholesterol-lowering Drug In U.S. Hisun, best known for a proposed JV with Pfizer on branded generics, is looking to move up the value chain into R&D. SHANGHAI – One of China’s leading active pharma-latory consulting for the IND preparation (CMC, phar-ceutical ingredient and finished product manufacturers macology/toxicology and clinical development plan/
CONFIDENTIAL HEALTH HISTORY I. CIRCLE APPROPRIATE ANSWER (Leave blank if you do not understand the question) Has there been a change in your health within the last year? Have you gone to the hospital or emergency room or had a serious illness in the last three years? Are you being treated by a physician now? If YES, explain Have you had problems with prior dental treatment? II. HA