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1.  Factors associated with the determination of antibiotic activity in bovine semen. 
Rosaramicin, an agent shown to be effective in vitro against ureaplasma of bovine origin was tested as an additive to bovine semen extender. Although some reduction in semen quality occurred it was still deemed satisfactory for use. In a test involving 41 cows inseminated once at estrus with rosaramicin-treated semen (162 mcg/mL) the nonreturn rate was 24% compared to a calculated average for this semen of 63% (n = 3310). The effect of centrifugation, time and temperature was examined in vitro using a combination of 150 mcg of lincomycin, 300 mcg of spectinomycin and 450 mcg of tylosin against ten strains of bovine ureaplasma. This combination has ureaplasmacidal activity and is suggested as an additive to semen extenders for the control of ureaplasma.
PMCID: PMC1235980  PMID: 6230144
2.  A comparison of the in vitro activity of two antibiotics against bovine ureaplasmas. 
A comparison of the in vitro activity of rosaramicin and minocin against 52 bovine Ureaplasma sp. strains, 20 of which were isolated from semen, 23 from vaginal swabs, and nine from preputial washings, has shown that both antibiotics have good inhibitory action against the strains tested. Rosaramicin was ureaplasmacidal in most instances at, or close to the inhibitory level while for most strains studied a ureaplasmacidal level of minocin was not found.
PMCID: PMC1320133  PMID: 7260728
3.  Antibiotic susceptibility profiles of Mycoplasma sp. 1220 strains isolated from geese in Hungary 
BMC Veterinary Research  2016;12:170.
Mycoplasma sp. 1220 can induce inflammation primarily in the genital and respiratory tracts of waterfowl, leading to serious economic losses. Adequate housing and appropriate antibiotic treatment are promoted in the control of the disease. The aim of the present study was to determine the in vitro susceptibility to thirteen different antibiotics and an antibiotic combination of thirty-eight M. sp. 1220 strains isolated from geese and a duck in several parts of Hungary, Central Europe between 2011 and 2015.
High MIC50 values were observed in the cases of tilmicosin (>64 μg/ml), oxytetracycline (64 μg/ml), norfloxacin (>10 μg/ml) and difloxacin (10 μg/ml). The examined strains yielded the same MIC50 values with spectinomycin, tylosin and florfenicol (8 μg/ml), while enrofloxacin (MIC50 5 μg/ml), doxycycline (MIC50 5 μg/ml), lincomycin (MIC50 4 μg/ml) and lincomycin-spectinomycin (1:2) combination (MIC50 4 μg/ml) inhibited the growth of the bacteria with lower concentrations. Tylvalosin (MIC50 0.5 μg/ml) and two pleuromutilins (tiamulin MIC50 0.625 μg/ml; valnemulin MIC50 ≤ 0.039 μg/ml) were found to be the most effective drugs against M. sp. 1220. However, strains with elevated MIC values were detected for all applied antibiotics.
Valnemulin, tiamulin and tylvalosin were found to be the most effective antibiotics in the study. Increasing resistance was observed in the cases of several antibiotics. The results highlight the importance of testing Mycoplasma species for antibiotic susceptibility before therapy.
PMCID: PMC4992293  PMID: 27543140
Antibiotic resistance; Duck; Goose; MIC; Microbroth dilution; Mycoplasma sp. 1220
4.  Susceptibilities of Mycoplasma bovis, Mycoplasma dispar, and Ureaplasma diversum strains to antimicrobial agents in vitro. 
The purpose of this study was to determine the susceptibility of various strains of Mycoplasma bovis, Mycoplasma dispar, and Ureaplasma diversum, which are prevalent causes of pneumonia in calves, to 16 antimicrobial agents in vitro. The MICs of the antimicrobial agents were determined by a serial broth dilution method for 16 field strains and the type strain of M. bovis, for 19 field strains and the type strain of M. dispar, and for 17 field strains of U. diversum. Final MICs for M. bovis and M. dispar were read after 7 days and final MICs for U. diversum after 1 to 2 days. All strains tested were susceptible to tylosin, kitasamycin, and tiamulin but were resistant to nifuroquine and streptomycin. Most strains of U. diversum were intermediately susceptible to oxytetracycline but fully susceptible to chlortetracycline; most strains of M. bovis and M. dispar, however, were resistant to both agents. Strains of M. dispar and U. diversum were susceptible to doxycycline and minocycline, but strains of M. bovis were only intermediately susceptible. Susceptibility or resistance to chloramphenicol, spiramycin, spectinomycin, lincomycin, or enrofloxacin depended on the species but was not equal for the three species. The type strains of M. bovis and M. dispar were more susceptible to various antimicrobial agents, including tetracyclines, than the field strains. This finding might indicate that M. bovis and M. dispar strains are becoming resistant to these agents. Antimicrobial agents that are effective in vitro against all three mycoplasma species can be considered for treating mycoplasma infections in pneumonic calves. Therefore, tylosin, kitasamycin, and tiamulin may be preferred over oxytetracycline and chlortetracycline.
PMCID: PMC187660  PMID: 8452363
5.  Ureaplasma urealyticum causing persistent urethritis in a patient with hypogammaglobulinaemia. 
Genitourinary Medicine  1985;61(6):404-408.
Ureaplasma urealyticum organisms (ureaplasmas) were isolated in large numbers (up to 10(8) colour changing units (ccu)/ml) over a period of four years from the urethra of a man with hypogammaglobulinaemia and non-gonococcal urethritis. Elimination of Mycoplasma hominis by antibiotic treatment early in the course of the urethritis did not diminish the severity of his condition, which indicated that this mycoplasma was not a cause. Courses of treatment with tetracyclines, spectinomycin, erythromycin, rosaramicin, and clindamycin on each occasion reduced the numbers of ureaplasma isolated from the urethra and the severity of disease. The organisms were not eliminated, however, sometimes due to the development of antibiotic resistance, and the urethritis recurred. Though netilmicin was not particularly effective in vitro, it was effective clinically, the disease resolving and the organisms disappearing for five months. Recurrence of urethritis, accompanied by epididymitis, was associated this time with the recovery of a different (tetracycline sensitive) ureaplasma strain; the urethritis and epididymitis were treated successfully with a combination of netilmicin and doxycycline. The administration of ureaplasma antiserum did not seem to be instrumental in eradicating the ureaplasmas. The various antibiotics had a greater influence on the mycoplasmas in the urethra than on those in the throat and joints, perhaps because the antibiotics were concentrated in the urogenital tract. The close association between the occurrence of urethritis and the ureaplasmas suggests strongly that they were responsible for it.
PMCID: PMC1011870  PMID: 4086030
6.  Control of Pleuropneumonia-like Organisms in Cell Culture 
Applied Microbiology  1967;15(6):1442-1446.
Mammalian cell culture systems were maintained free of mycoplasmas by using a 3-day agar plate test as a weekly routine to monitor the conditions of the cells. If contaminated cell cultures were found, they were discarded and replaced from a pleuropneumonia-like organism (PPLO)-free cell bank. PPLO-free lines were established by treatment with various antibiotics. The KB cell line was freed of mycoplasmas by treatment for 1 week with a mixture of chlortetracycline, kanamycin, and chloramphenicol. L-929 cells were cleared of contamination with either spectinomycin or tylosin, and a synovial cell line was cleared with lincomycin or tylosin. Each cell line, after eradication of the contaminant, was stored in liquid nitrogen. A number of agents were tested to determine minimal inhibitory concentration against three known and three unidentified mycoplasmas. Chlortetracycline and tetracycline were found to be highly active against all strains, whereas tylosin, spectinomycin, and lincomycin, though less active, were equally useful because of their low toxicity against cells. Kanamycin was highly active against three strains, but inactive at high levels against the KB cell contaminants. A disc plate test was used to check isolated cell contaminants for sensitivity to various agents.
PMCID: PMC547226  PMID: 16349761
7.  Comparison of methods for in vitro testing of susceptibility of porcine Mycoplasma species to antimicrobial agents. 
The MICs of 18 antimicrobial agents used against strains of three porcine Mycoplasma species were determined by a serial broth dilution method. Twenty field strains of M. hyorhinis, ten field strains of M. hyopneumoniae, six field strains of M. flocculare, and the type strains of these species were tested. Twelve field strains and the type strain of M. hyorhinis were also tested by an agar dilution method. Tests were read at various time points. When the broth dilution method was used, the final MIC had to be read 2 days after color changes had stopped. MICs of tetracycline, oxytetracycline, doxycycline, and minocycline were low for the three Mycoplasma species tested. MICs of chlortetracycline were 8 to 16 times higher than MICs of the other tetracyclines. Spiramycin, tylosin, kitasamycin, spectinomycin, tiamulin, lincomycin, and clindamycin were effective against all strains of M. hyorhinis and M. hyopneumoniae. The quinolones were highly effective against M. hyopneumoniae but less effective against M. hyorhinis. The susceptibility patterns for M. hyopneumoniae and M. flocculare were similar.
PMCID: PMC244982  PMID: 2024954
8.  Antimicrobial Activities of 81.723 hfu, a New Pleuromutilin Derivative 
The new pleuromutilin derivative 81.723 hfu is extremely active against gram-positive organisms such as streptococci, staphylococci, and against mycoplasmas. A number of Shigella, Klebsiella, and Escherichia coli strains were also found to be quite susceptible to this new agent, whereas other gram-negative organisms like Pseudomonas aeruginosa, Proteus species, and Alcaligenes faecalis proved to be naturally resistant to 81.723 hfu. The new compound acts bacteriostatically. Bactericidal effects have been observed only at concentrations which are 100-fold higher than the minimal inhibitory concentrations. The new antibiotic is well tolerated in all animal species tested so far and has been successfully used in the treatment of experimental infections with gram-positive organisms and with mycoplasmas in mice and rats. Resistance against this new compound arose gradually in all microorganisms investigated. It is noteworthy that the rate at which resistance against 81.723 hfu emerged in mycoplasmas (Mycoplasma gallisepticum and Mycoplasma hyorhinis) was significantly slower than the corresponding rate at which resistance against tylosin tartrate appeared. Mycoplasma strains which became insensitive to 81.723 hfu were also resistant to tylosin tartrate, whereas mycoplasmas which developed resistance against tylosin tartrate, although less sensitive to 81.723 hfu than wild-type strains, were still eliminated by this drug. In a strain of Klebsiella pneumoniae, complete cross-resistance was observed between the pleuromutilin derivative on one hand and lincomycin and erythromycin on the other. Modest degrees of cross-resistance were also observed with chloramphenicol. However, it appears unlikely that the latter phenomenon is sufficiently pronounced to affect treatment with either antibiotic.
PMCID: PMC429174  PMID: 1170807
9.  Comparisons of antibiotic combinations to control Pseudomonas aeruginosa in bovine semen. 
Raw semen experimentally contaminated with 10(6) Pseudomonas aeruginosa cells per milliliter was processed for use in artificial insemination (AI) using three different antibiotic combinations: a) gentamicin, lincomycin, spectinomycin and tylosin (GLST) directly added to contaminated raw semen followed by dilution with whole milk or egg yolk Tris containing GLST; b) penicillin, streptomycin, lincomycin, spectinomycin and minocycline (PSLSM) in whole milk used to dilute the contaminated raw semen followed by further dilution with glycerolated milk containing PSLSM; and c) penicillin, streptomycin, lincomycin and spectinomycin (PSLS) used with egg yolk Tris diluent in the same way as PSLSM and milk. Diluted semen was incubated at 35 degrees C for 5 or 40 min before cooling commenced. To assess the efficacy of the antibiotics in controlling P. aeruginosa, diluted semen samples were cultured for the organism before and after freezing. The GLST antibiotics added to raw semen and milk reduced the counts of P. aeruginosa before or after freezing. When egg yolk Tris was used, GLST inhibited the organism as indicated by its low growth in culture before freezing and absence of growth from samples after freezing. With PSLSM and PSLS treatments, the organism was recovered in milk and egg yolk Tris processed semen both before and after freezing. However, incubation at 35 degrees C for 40 min prior to cooling, compared to incubation of 5 min, appeared to reduce the bacterial counts after freezing.
PMCID: PMC1263738  PMID: 7704847
10.  In vitro antimicrobial activity of rosoxacin against Neisseria gonorrhoeae, Chlamydia trachomatis, and Ureaplasma urealyticum. 
The antimicrobial activity of rosoxacin, a new quinoline antibacterial compound, was determined against the causative organisms of three sexually transmitted diseases. Rosoxacin demonstrated a high degree of activity against Neisseria gonorrhoeae clinical isolates, with the minimal inhibitory concentrations for 50% of these being 0.03 microgram/ml. The corresponding minimal inhibitory concentrations for penicillin, ampicillin, tetracycline, and spectinomycin were 0.25 U/ml, 0.125 microgram/ml, 0.25 microgram/ml, and 16 microgram/ml, respectively. Eleven strains of Chlamydia trachomatis were inhibited by 5 microgram of rosoxacin per ml, and each of seven Ureaplasma urealyticum strains was inhibited by 2 to 8 microgram of rosoxacin per ml. The results of these susceptibility studies, coupled with those of an earlier evaluation of the pharmacokinetics of rosoxacin, provide support for extending or undertaking clinical evaluations of this compound against infections with N. gonorrhoeae, C. trachomatis, and U. urealyticum.
PMCID: PMC284084  PMID: 6778385
11.  Antibiotic susceptibility profiles of Mycoplasma bovis strains isolated from cattle in Hungary, Central Europe 
BMC Veterinary Research  2014;10:256.
Mycoplasma bovis is a worldwide pathogen, causative agent of pneumonia, mastitis, arthritis, and a variety of other symptoms in cattle. The economic losses due to mycoplasma pneumonia could be reduced by antibiotic treatment. The aim of the present study was to determine the in vitro susceptibility of M. bovis strains isolated from cattle in Hungary to eleven antibiotics.
Minimal inhibitory concentration (MIC) values of 35 M. bovis strains collected from different parts of Hungary between 2010 and 2013 were determined by the microbroth dilution method. Strains with high MIC values were found in the case of all applied antibiotics. The most effective antibiotics tested in vitro were fluoroquinolones (MIC90 danofloxacin 0.312 μg/ml, enrofloxacin 0.312 μg/ml, marbofloxacin 0.625 μg/ml). Our results confirm the observations of increasing MIC values to antibiotics commonly used in the therapy of mycoplasma infections, primarily to tetracyclines; tetracycline (MIC90 16 μg/ml) and oxytetracycline (MIC90 ≥ 64 μg/ml) and macrolides; tylosin (MIC90 ≥ 128 μg/ml) and tilmicosin (MIC90 ≥ 128 μg/ml). The growth of many M. bovis strains was not inhibited by gentamicin (MIC90 8 μg/ml), spectinomycin (MIC90 ≥ 256 μg/ml), florfenicol (MIC90 8 μg/ml) or lincomycin (MIC90 ≥ 64 μg/ml).
Our results emphasize the necessity of periodic testing for antibiotic susceptibility in this geographic region. Based on our in vitro examinations, fluoroquinolones could be the most effective drugs for the therapy of M. bovis infections in Hungary. However, current antimicrobial use policies have to be taken into account to avoid further antibiotic resistance development and to reserve fluoroquinolones for the treatment of severe infections which have responded poorly to other classes of antimicrobials.
PMCID: PMC4210529  PMID: 25344297
Antibiotic resistance; MIC; Fluoroquinolones; Microbroth dilution; Mycoplasma bovis
12.  In vitro and in vivo activities of sedecamycin against Treponema hyodysenteriae. 
Sedecamycin (lankacidin A), one of the lankacidin-group antibiotics, showed potent activity against Treponema hyodysenteriae. The MICs of sedecamycin against 79 field isolates of T. hyodysenteriae ranged from 0.78 to 12.5 micrograms/ml, the MIC for 90% of the strains tested (MIC90) being 3.13 micrograms/ml. The protective and therapeutic effects of sedecamycin were compared with those of carbadox, tiamulin, and lincomycin against experimental infection with T. hyodysenteriae in mice. The protective effect of sedecamycin was similar to that of carbadox, two times more potent than that of tiamulin, and three times greater than that of lincomycin. In the therapeutic test, sedecamycin showed activity similar to that of carbadox and was two times more active than both tiamulin and lincomycin. At doses of 10 mg or more of sedecamycin per kg, the recurrence of shedding of T. hyodysenteriae into the feces of mice was not detected for at least 8 weeks postmedication.
PMCID: PMC172201  PMID: 3377458
13.  Antimicrobial susceptibility of ileal symbiont intracellularis isolated from pigs with proliferative enteropathy. 
Journal of Clinical Microbiology  1995;33(5):1314-1317.
Proliferative enteropathy is caused by the microaerophilic obligate intracellular bacterium ileal symbiont (IS) intracellularis. Treatment of this disease is problematic because of the lack of in vivo or in vitro data on the activities of antimicrobial agents. A new procedure for determining the susceptibility of IS intracellularis was developed by using a tissue culture system which promotes the in vitro multiplication of this organism. Nineteen antimicrobial agents were evaluated in triplicate cultures for their intracellular and extracellular activities against up to three IS intracellularis strains isolated from pigs with proliferative enteropathy. The MIC was defined as the lowest concentration which prevented multiplication of 99% of the IS intracellularis isolates. Penicillin, erythromycin, difloxacin, virginiamycin, and chlortetracycline were the most active compounds tested, all with MICs of < or = 1 microgram/ml. Tiamulin and tilmicosin were the next most active compounds, with MICs of < or = 4 micrograms/ml. The MICs of aminoglycosides were generally > 32 micrograms/ml. Both lincomycin and tylosin were relatively inactive against the IS intracellularis strains tested, with MICs of 32 and 64 micrograms/ml, respectively. These results indicate that some compounds capable of intracytoplasmic accumulation and blocking bacterial protein synthesis were active against IS intracellularis strains isolated from pigs with proliferative enteropathy. The in vitro cultivation system shows promise as a method for studying the interaction between IS intracellularis and antimicrobial agents and for screening new antibiotics for use in therapy.
PMCID: PMC228152  PMID: 7615747
14.  Comparison of MICs of ceftiofur and other antimicrobial agents against bacterial pathogens of swine from the United States, Canada, and Denmark. 
Journal of Clinical Microbiology  1995;33(9):2435-2444.
The MICs of ceftiofur and other antimicrobial agents, tested for comparison, for 515 bacterial isolates of pigs from the United States, Canada, and Denmark with various diseases were compared. The organisms tested included Actinobacillus pleuropneumoniae, Escherichia coli, Pasteurella multocida, Salmonella choleraesuis, Salmonella typhimurium, Streptococcus suis, Streptococcus dysgalactiae subsp. equisimilis, Streptococcus equi subsp. equi, and Streptococcus equi subsp. zooepidemicus. In addition to ceftiofur, the following antimicrobial agents or combinations were tested: enrofloxacin, ampicillin, sulfamethazine, trimethoprim-sulfadiazine (1:19), erythromycin, lincomycin, spectinomycin, lincomycin-spectinomycin (1:8), tilmicosin, and tetracycline. Tilmicosin was only tested against the U.S. isolates. Overall, ceftiofur and enrofloxacin were the most active antimicrobial agents tested against all isolates, with MICs inhibiting 90% of isolates tested (MIC90s) of < or = 2.0 and < or = 1.0 microgram/ml, respectively. Erythromycin, sulfamethazine, spectinomycin, and lincomycin demonstrated limited activity against all of the organisms tested, with MIC90s of > or = 8.0, > or = 256.0, > or = 32.0, and > or = 16.0 micrograms/ml, respectively. Trimethoprim-sulfadiazine was active against isolates of A. pleuropneumoniae, S. choleraesuis, S. typhimurium, P. multocida, S. equi, and S. suis (MIC90s, < or = 0.5 microgram/ml) but was less active against the E. coli strains tested (MIC90, > 16.0 micrograms/ml). Ampicillin was active against the P. multocida, S. suis, and S. equi isolates tested (MIC90s, 0.5, 0.06, and 0.06 micrograms/ml, respectively) and was moderately active against S. typhimurium (MIC90s, 2.0 micrograms/ml). However, this antimicrobial agent was much less active when it was tested against A. pleuropneumoniae, S. cholerae-suis, and E. coli (MIC90s, 16.0, > 32.0, and 32.0 micrograms/ml, respectively). Against the U.S. isolates of A. pleuropneumoniae and P. multocida, tilmicosin was moderately active (MIC90s, 4.0 and 8.0 micrograms/ml, respectively). However, this compound was not active against the remaining U.S. isolates (MIC90s, > 64.0 micrograms/ml). Differences in the MICs from one country to another were not detected with enrofloxacin, ceftiofur, or lincomycin for the strains tested, but variations in the MICs of the remaining antimicrobial agents were observed.
PMCID: PMC228432  PMID: 7494042
15.  Pharmacokinetic/Pharmacodynamic Profiles of Tiamulin in an Experimental Intratracheal Infection Model of Mycoplasma gallisepticum 
Mycoplasma gallisepticum is the most important pathogen in poultry among four pathogenic Mycoplasma species. Tiamulin is a pleuromutilin antibiotic that shows a great activity against M. gallisepticum and has been approved for use in veterinary medicine particularly for poultry. However, the pharmacokinetic/pharmacodynamics (PK/PD) profiles of tiamulin against M. gallisepticum are not well understood. Therefore, in the current studies, we investigated the in vivo PK/PD profiles of tiamulin using a well-established experimental intratracheal infection model of M. gallisepticum. The efficacy of tiamulin against M. gallisepticum was studied in 8-day-old chickens after intramuscular (i.m.) administration at 10 doses between 0–80 mg/kg. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to evaluate the PK parameters of tiamulin following i.m. administration at doses of 5, 40, and 80 mg/kg in Mycoplasma gallisepticum-infected neutropenic chickens. Real-time PCR (RT-PCR) was used for quantitative detection of M. gallisepticum. The MIC of tiamulin against M. gallisepticum strain S6 was 0.03 μg/mL. The PK/PD index, AUC24h/MIC, correlated well with the in vivo antibacterial efficacy. The in vivo data suggest that animal dosage regimens should supply AUC24h/MIC of tiamulin of 382.68 h for 2 log10 ccu equivalents M. gallisepticum reduction. To attain that goal, the administered dose is expected to be 45 mg/kg b.w. for treatment of M. gallisepticum infection with an MIC90 of 0.03 μg/mL.
PMCID: PMC5012102  PMID: 27656647
tiamulin; M. gallisepticum; in vivo PK/PD; chicken
16.  Susceptibilities of genital mycoplasmas to the newer quinolones as determined by the agar dilution method. 
The increasing resistance of genital mycoplasmas to tetracycline poses a problem because tetracycline is one of the few antimicrobial agents active against Mycoplasma hominis, Ureaplasma urealyticum, chlamydiae, gonococci, and other agents of genitourinary-tract disease. Since the quinolones are a promising group of antimicrobial agents, the susceptibilities of M. hominis and U. urealyticum to the newer 6-fluoroquinolones were determined by the agar dilution method. Ciprofloxacin, difloxacin, and ofloxacin had good activity against M. hominis, with the MIC for 50% of isolates tested (MIC50) being 1 microgram/ml. Fleroxacin, lomefloxacin, pefloxacin, and rosoxacin had MIC50s of 2 micrograms/ml. Enoxacin, norfloxacin, and amifloxacin had MIC50s of 8 to 16 micrograms/ml, and cinoxacin and nalidixic acid were inactive (MIC50, greater than or equal to 256 micrograms/ml). Overall, the activities of 6-fluoroquinolones for ureaplasmas were similar to those for M. hominis, with MICs being the same or twofold greater. The most active 6-fluoroquinolones against ureaplasmas were difloxacin, ofloxacin, and pefloxacin, with MIC50s of 1 to 2 micrograms/ml. Ciprofloxacin was unusual in that the MIC50 for M. hominis was 1 microgram/ml, whereas the MIC50 for ureaplasmas was 8 micrograms/ml. Since the MIC50s for the most active quinolones approximate achievable concentrations in blood and urine, quinolones have promise in treating mycoplasmal infections.
PMCID: PMC171429  PMID: 2712541
17.  Antimicrobial susceptibility of porcine Brachyspira hyodysenteriae and Brachyspira pilosicoli isolated in Sweden between 1990 and 2010 
The anaerobic spirochetes Brachyspira hyodysenteriae and Brachyspira pilosicoli cause diarrheal diseases in pigs. Their fastidious nature has hampered standardization of methods for antimicrobial susceptibility testing. For monitoring of antimicrobial susceptibility wild type cutoff values are needed to define where the wild type distribution of MICs ends and no approved cutoffs are available for Brachyspira spp. In this study antimicrobial susceptibility data for both species (in total 906 isolates) were compiled and analyzed and wild type cut off values for B. hyodysenteriae proposed.
The MICs of tiamulin, valnemulin, tylosin, tylvalosin, doxycycline and lincomycin were determined by broth dilution in brain heart infusion broth supplemented with 10% fetal calf serum.
The compiled MICs from the broth dilution tests of the B. hyodysenteriae type strain, B78T (ATCC® 27164T), showed that the method yields reproducible results. In an international perspective the frequencies of isolates with decreased antimicrobial susceptibility were low among both B. hyodysenteriae and B. pilosicoli. However, in B. pilosicoli a constant level of 10-15% isolates with tiamulin MICs >4 μg/ml was detected between 2002 and 2010 and in B. hyodysenteriae a gradual increase in tiamulin MICs was seen between 1990 and 2003 although this increase has ceased during the last years. The wild type cutoff values proposed for B. hyodysenteriae are: tiamulin >0.25 μg/ml, valnemulin >0.125 μg/ml, tylosin >16 μg/ml, tylvalosin >1 μg/ml, lincomycin >1 μg/ml and doxycycline >0.5 μg/ml.
The broth dilution method used in this study has over the years generated tightly grouped MIC populations for the field isolates and reproducible results for the control strain B78T and is therefore a suitable antimicrobial susceptibility test method for monitoring of Brachyspira spp. Here we propose wild type cutoff values for six antimicrobial agents for B. hyodysenteriae tested by broth dilution based on MIC distributions and the current knowledge on mechanisms of resistance in this species. There are few studies on antimicrobial resistance mechanisms and MIC distributions in B. pilosicoli but to some extent the cutoff values proposed for B. hyodysenteriae may be applicable also for monitoring of antimicrobial susceptibility in B. pilosicoli.
PMCID: PMC3526423  PMID: 22998753
Brachyspira hyodysenteriae; Swine dysentery; Brachyspira pilosicoli; Porcine intestinal spirochetosis; Antimicrobial susceptibility
18.  A study on the effect of Pseudomonas aeruginosa in semen on bovine fertility. 
Two experiments were done to demonstrate whether the presence of Pseudomonas aeruginosa in bovine semen could affect fertilization and/or early embryonic development. In the first experiment, superovulated heifers were inseminated with semen naturally contaminated with P. aeruginosa (ADRI 102) or clean semen and seven day-old embryos were collected nonsurgically. The endometrium of treated heifers appeared more sensitive to the flush procedures. In experiment 2, heifers were inseminated at synchronized estrus with semen experimentally contaminated with P. aeruginosa (ADRI 102) and processed in the same way as commercial semen with antibiotics (gentamicin, lincomycin, spectinomycin and tylosin) or processed without antibiotics added. Embryos were recovered at slaughter seven days later. In general, there was no significant reduction in fertility or development of embryos in vitro as a result of relatively high numbers of P. aeruginosa in bovine semen.
PMCID: PMC1263739  PMID: 7704848
19.  In vitro activities of U-63366, a spectinomycin analog; roxithromycin (RU 28965), a new macrolide antibiotic; and five quinolone derivatives against Haemophilus ducreyi. 
The in vitro activities of the new spectinomycin analog U-63366, the new macrolide roxithromycin (RU 28965), and five new quinolone derivatives (pefloxacin, rosoxacin, norfloxacin, ofloxacin, and ciprofloxacin) were studied against 23 multiresistant strains of Haemophilus ducreyi (beta-lactamase producers) isolated in Paris and were compared with the activities of tetracycline, minocycline, chloramphenicol, streptomycin, kanamycin, gentamicin, spectinomycin, erythromycin, and nalidixic acid. All strains were uniformly susceptible to the seven new antibiotics tested. Ciprofloxacin had the greatest inhibitory effect in vitro (the MIC for 90% of the strains tested [MIC90] was 0.016 microgram/ml), and U-63366 was the most active aminoglycoside-aminocyclitol antibiotic (MIC90, 0.25 microgram/ml).
PMCID: PMC180592  PMID: 2946262
20.  Effects of Increasing Concentrations of Minocin on the Motility of Bovine Spermatozoa Processed in Various Extenders 
The Canadian Veterinary Journal  1979;20(9):233-236.
The effect of minocycline hydrochloride (Minocin) on bovine spermatozoa was studied in eggyolk-citrate, Tris buffer and whole milk extenders. Each of the extenders contained penicillin, streptomycin, lincomycin-spectinomycin and four levels of minocin (10, 260, 510 and 760 μg/ml). Split ejaculates from Holstein sires were diluted in each of the extenders. This was followed by microscopic evaluation for progressive motility after initial dilution at 32°C, after cooling at 5°C and complete dilution, immediately after freezing and after 14 days of storage in liquid nitrogen. Under the experimental conditions employed, a decrease in the percent progressive motility was observed in the eggyolk-citrate and Tris buffer extenders with increasing concentration of minocin. Little or no difference in the percent progressive motility in the whole milk extender, either before or after freezing was observed when the minocin concentration was elevated.
PMCID: PMC1789589  PMID: 508386
21.  The current epidemiological status of infectious coryza and efficacy of PoulShot Coryza in specific pathogen-free chickens 
Journal of Veterinary Science  2016;17(3):323-330.
Infectious coryza (IC) is an infectious disease caused by Avibacterium (Av.) paragallinarum. IC is known to cause economic losses in the poultry industry via decreased egg production in layers. Between 2012 and 2013, Av. paragallinarum was isolated from seven chicken farms by Chungbuk National University. We identified Av. paragallinarum, the causative pathogen of IC by polymerase chain reaction (PCR) and serovar serotype A, by multiplex PCR. Antibiotic sensitivity tests indicated that a few field-isolated strains showed susceptibility to erythromycin, gentamicin, lincomycin, neomycin, oxytetracycline, spectinomycin, and tylosin. A serological survey was conducted to evaluate the number of flocks that were positive for Av. paragallinarum by utilizing a HI test to determine the existence of serovar A. Serological surveys revealed high positivity rates of 86.4% in 2009, 78.9% in 2010, 70.0% in 2011, and 69.6% in 2012. We also challenged specific pathogen-free chickens with isolated domestic strains, ADL121286 and ADL121500, according to the measured efficacy of the commercial IC vaccine, PoulShot Coryza. We confirmed the effectiveness of the vaccine based on relief of clinical signs and a decreased re-isolation rate of ADL121500 strain. Our results indicate IC is currently prevalent in Korea, and that the commercial vaccine is effective at protecting against field strains.
PMCID: PMC5037299  PMID: 26726018
Avibacterium paragallinarum; infectious coryza; infectious coryza vaccine; serologic survey
22.  Ureaplasma urealyticum, Ureaplasma parvum, Mycoplasma hominis and Mycoplasma genitalium infections and semen quality of infertile men 
Genital ureaplasmas (Ureaplasma urealyticum and Ureaplasma parvum) and mycoplasmas (Mycoplasma genitalium and Mycoplasma hominis) are potentially pathogenic species playing an etiologic role in both genital infections and male infertility. Reports are, however, controversial regarding the effects of these microorganisms infections in the sperm seminological variables. This study aimed at determining the frequency of genital ureplasmas and mycoplasmas in semen specimens collected from infertile men, and at comparing the seminological variables of semen from infected and non-infected men with these microorganisms.
A total of 120 semen samples collected from infertile men were investigated. Semen specimens were examined by in-house PCR-microtiter plate hybridization assay for the presence of genital ureaplasmas and mycoplasmas DNA. Semen analysis was assessed according to the guidelines of the World Health Organization. Standard parametric techniques (t-tests) and nonparametric techniques (Wilcoxon tests) were used for statistical analysis.
The frequency of genital ureaplasmas and mycoplasmas detected in semen samples of infertile men was respectively 19.2% (23/120) and 15.8% (19/120). The frequency of Ureaplasma urealyticum (15%) was higher than that of Mycoplasma hominis (10.8%), Ureaplasma parvum (4.2%) and Mycoplasma genitalium (5%). Mixed species of mycoplasmas and ureaplasmas were detected in 6.7% of semen samples.
Comparison of the parameters of the standard semen analysis between the male partners of the infertile couples with and without genital ureaplasmas and mycoplasmas infection showed that the presence of Mycoplasma hominis DNA in semen samples is associated with low sperm concentration (p = 0.007) and abnormal sperm morphology (p = 0.03) and a negative correlation between sperm concentration and the detection of Mycoplasma genitalium in semen samples of infertile men (p = 0.05). The mean values of seminal volume, pH, vitality, motility and leukocyte count were not significantly related either to the detection of genital mycoplasmas DNA or to the detection of ureaplasmas DNA in semen specimens.
Our results demonstrate that genital mycoplasmas and ureaplasmas seem to be widespread among the male partners of infertile couples in Tunisia. Genital mycoplasmas infections of the male genital tract could negatively influence semen quality. Our results also indicate that PCR-microtiter plate hybridization assay method provides a rapid and effective technique to detect human genital mycoplasmas and ureaplasmas which is useful for etiological and epidemiological studies of these pathogens.
PMCID: PMC2194714  PMID: 17988404
23.  Comparative susceptibilities of various animal-pathogenic mycoplasmas to fluoroquinolones. 
The in vitro activities of six antimicrobial agents were tested against 162 mycoplasma strains of eight species isolated from poultry and livestock at different geographic sites. Tiamulin was most active (MICs at which 90% of the isolates were inhibited [MIC90s], 0.025 to 0.25 microg/ml); enrofloxacin and danofloxacin had near equivalent activities (MIC90s, 0.05 to 1.0 microg/ml), but were much more active than flumequine (MIC90s, 1 to 50 microg/ml). The MIC90s of tylosin and oxytetracycline were 0.25 to > 100 microg/ml and 0.25 to 100 microg/ml, respectively.
PMCID: PMC164063  PMID: 9303412
24.  Minimal inhibitory concentrations of antimicrobial agents against Actinobacillus pleuropneumoniae. 
Forty-five isolates of Actinobacillus pleuropneumoniae were tested for susceptibility to 12 antimicrobial agents using a microdilution method for the minimal inhibitory concentration determinations. These results confirmed the high prevalence of A. pleuropneumoniae strains resistant to antibiotics as reported earlier using the disc diffusion method (Kirby-Bauer method). While 36% of the isolates were resistant to the penicillins, 47% were resistant to chloramphenicol and 68% were resistant to tetracycline. Minimal inhibitory concentrations for the resistant isolates were approximately 32 times higher than those for the susceptible isolates to the above antibacterial agents. The isolates were in general weakly susceptible or resistant to spectinomycin, lincomycin, tiamulin and spiramycin whereas most of them were susceptible to gentamicin, trimethoprim and erythromycin. The susceptibility pattern was similar throughout the 1980 to 1984 period. The 14 serotype 5 isolates were more resistant to tetracycline but less resistant to chloramphenicol and the penicillins than the 28 serotype 1 isolates.
PMCID: PMC1255455  PMID: 3167716
25.  Comparison of antimicrobial susceptibility patterns of Campylobacter jejuni and Campylobacter coli. 
To determine whether employing antibiograms is useful to separate Campylobacter jejuni and Campylobacter coli, we determined the MICs of 12 antibiotics for 104 human clinical strains and 74 swine strains. Of 74 swine strains, 5 (7%) were hippurate positive, as were 93 (89%) of 104 human strains. The 12 antimicrobial agents tested were ampicillin, amoxicillin, clindamycin, chloramphenicol, erythromycin, furazolidone, norfloxacin, nalidixic acid, rosoxacin, rosaramicin, tetracycline, and Sch 32063. Isolates from humans were significantly (P less than 0.001) more susceptible than swine strains to clindamycin, erythromycin, rosaramicin, and Sch 32063. Of 11 human hippurate-negative strains, 3 (27%) were resistant to clindamycin, erythromycin, rosaramicin, and Sch 32063, compared with 1 of 93 (1%) hippurate-positive strains. Nearly all human and swine strains were susceptible to furazolidone and nalidixic acid. Campylobacter isolates from humans and swine have different antibiograms, and the susceptibility to certain antibiotics, such as clindamycin, may be helpful for differentiation of C. jejuni from C. coli.
PMCID: PMC176168  PMID: 6508265

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