In vitro action of combinations of antimicrobial agents and EDTA-tromethamine on Pseudomonas aeruginosa

Combinations of EDTA-tromethamine and 7 antimicrobial agents (chloramphenicol, nalidixic acid, oxytetracycline, penicillin, polymyxin-B, streptomycin, and triple sulfa) were tested for synergistic activities against Pseudomonas aeruginosa. Three in vitro tests were used, including minimal inhibitory concentrations of the drugs, a 2-dimensional Microtiter checkerboard technique, and bacterial inhibition studies. A synergistic inhibitory action was observed with combinations of EDTA-tromethamine plus penicillin and EDTA-tromethamine plus oxytetracycline. When chloramphenicol, streptomycin, nalidixic acid, polymyxin-B, or triple sulfa was mixed with EDTA-tromethamine, synergistic action did not occur.     

Potentiation of antibiotic activity by EDTA-tromethamine against three clinically isolated Gram-positive resistant bacteria. Anin vitro investigation

Thein vitro synergistic effects of combinations of EDTA-tromethamine and five antimicrobial agents (ampicillin, cephalexin, oxytetracycline, streptomycin and sulphadimethoxine) on three clinically isolated Gram-positive bacteria (Staphylococcus aureus, Staphylococcus hominis andStreptococcus faecium) were investigated. The bacteria had been isolated from three cases of canine otitis resistant to -lactam antibiotic therapy. The antimicrobial activity was evaluated by measuring the minimal inhibitory concentration for the antibiotics alone or in combination with EDTA-tromethamine. EDTA-tromethamine potentiated the activity of cefalexin againstS. aureus andS. hominis, of oxytetracycline againstS. aureus andS. faecium and of streptomycin againstS. faecium. No significant effects were noted on the activity of oxytetracycline againstS. hominis. The remaining combinations gave a slight synergistic effect. As previously shown for Gram-negative resistant bacteria, these data suggest that the association of EDTA-tromethamine and appropriate antibiotic therapy may be useful to overcome persistent infections of soft tissues in domestic animals.Abbreviations cfu colony forming units - EDTA ethylenediaminetetraacetic acid - MIC minimal inhibitory concentration - tromethamine tris(hydroxymethyl)- aminomethane Part of this paper was communicated at the XLVI Congress of the Italian Society of Veterinary Sciences, Venice, 30 September – 3 October 1992     

In vitro effect of combinations of antimicrobial agents and EDTA-tromethamine on certain gram-positive bacteria

Combinations of EDTA-tromethamine and 7 antimicrobial agents (chloramphenicol, nalidixic acid, oxytetracycline, penicillin, polymyxin-B, streptomycin, and a triple sulfonamide preparation) were tested for synergistic activities against Staphylococcus aureus, Corynebacterium renal, Listeria monocytogenes, Erysipelothrix rhusiopathiae, and a beta-hemolytic streptococci. Two in vitro tests were used--minimal inhibitory concentrations of the drugs and a 2-dimensional Microtiter checkerboard technique. A slight synergistic action was seen when L monocytogenes was exposed to combinations of penicillin-EDTA or streptomycin-EDTA.     

Antibacterial action of combinations of oxytetracycline, dimethyl sulfoxide, and EDTA-tromethamine on Proteus, Salmonella, and Aeromonas

Antibacterial effects against Proteus mirabilis, Salmonella typhimurium, and Aeromonas hydrophila were obtained with subminimal inhibitory concentrations of oxytetracycline and EDTA-tromethamine. Antibacterial effects were not observed with subminimal inhibitory concentrations of dimethyl sulfoxide plus oxytetracycline or with dimethyl sulfoxide plus EDTA-tromethamine. Using a 2-dimensional Microtiter checkerboard technique, inhibitory activities of the various combinations of solutions were studied, and isobolograms were plotted. A synergistic effect was seen with combinations of oxytetracycline and EDTA-tromethamine. The greatest synergistic effect was observed when the mixture was caused to react with P mirabilis. These findings were confirmed by kinetic studies of microbial death, using one-fourth minimal inhibitory concentrations of these preparations.     

Inhibitory effects of combinations of oxytetracycline, dimethyl sulfoxide, and EDTA-tromethamine on Escherichia coli

Antibacterial activity against Escherichia coli was obtained with subminimal inhibitory concentrations of oxytetracycline (OTC) and EDTA-tromethamine. Inhibitory effects were not observed using combinations of dimethyl sulfoxide and OTC or dimethyl sulfoxide and EDTA-tromethamine. Neither EDTA-tromethamine nor OTC used alone was capable of the same degree of inhibition. Using a 2-dimensional Microtiter checkerboard technique, the inhibitory activity of these combinations was studied and isobolograms were plotted. A synergistic effect was seen with combinations of OTC and EDTA-tromethamine. Kinetic studies of microbial death, using subminimal inhibitory concentrations of these agents, confirmed these findings.     

Salmonella isolates from cloacal swabs and footpads of wild birds in the immediate environment of Tokyo Bay

A total of 328 cloacal swabs and 163 footpads of wild birds were investigated for the presence of salmonellae. All 19 isolates from cloacal swabs were serotyped as Salmonella Typhimurium susceptible to all five conventional antimicrobial agents (ampicillin, chloramphenicol, streptomycin, oxytetracycline and nalidixic acid) tested. In contrast, 15 salmonellae isolated from footpads included S. Muenhen, S. Virchow, S. Bareily and S. Bovismorbificans, including S. Typhimurium; these non-Salmonella Typhimurium isolates showed multiple drug resistance.     

Incidence of drug resistance and transmissible R factors in strains of E. coli isolated from faeces of healthy pigs

Two hundred and twenty-six strains of E. coli were isolated from faeces of 107 pigs at different ages and without clinical signs of infectious diseases. The resistance of the strains to sulphonamide, tetracycline, streptomycin, chloramphenicol, ampicillin, and nalidixic acid was determined. In 74 % of the animals the predominant E. coli flora was found to be resistant to one or more of the drugs mentioned. Fifty-three % of the strains were resistant. Multiple resistance was predominant among resistant strains (67 %). R factors transmissible to a sensitive strain of E. coli K12 W3132 were demonstrated in 28 %. The proportion of resistant strains was largest in young animals (0–14 weeks) accounting for 65 % of the strains isolated, as compared to 43 % of strains from pigs and sows (6 months or more). The incidence of resistance to sulphonamide, tetracycline, and streptomycin was high, whereas most of the strains were sensitive to ampicillin and chloramphenicol. All strains were sensitive to nalidixic acid.The incidence of resistance to antibiotics in a population of pigs to whom these drugs are not fed but applied as therapeutic agents solely seems rather high. When based on clinical findings only, the value is therefore questionable of sulphonamide, tetracycline and streptomycin treatment of infectious diseases caused by E. coli.     

In vitro safety assessments and antimicrobial activities of Lactobacillus rhamnosus strains isolated from a fermented mare's milk

Safety and probiotic characteristics such as antimicrobial activities of three Lactobacillus rhamnosus strains, FSMM15, FSMM22 and FSMM26, previously isolated as potential probiotics from fermented mare's milk were investigated. The three FSMM strains were susceptible to ampicillin, gentamycin, kanamycin, streptomycin, tetracycline and chloramphenicol, whereas they were resistant to erythromycin (minimal inhibitory concentration (MIC) = 4?8 µg/mL) and clindamycin (MIC = 4 µg/mL); bioconversion of bile salts, hemolytic activity and mucin degradation activity were negative; enzymatic activities of α‐chymotrypsin and β‐glucosidase were detected, but those of α‐galactosidase, β‐glucuronidase and N‐acetyl‐β‐glucosaminidase, were undetectable. Among the strains, strain FSMM15 was chosen as a safer probiotic candidate due mainly to the lack of plasminogen binding ability. Despite lower acid production of strain FSMM15 than others, its cell‐free culture supernatant inhibited growths of Salmonella Typhimurium LT‐2, Shigella sonnei , Listeria monocytogenes , and Escherichia coli O157 with comparable levels of ampicillin, suggesting a favorable aspect of strain FSMM15 as a probiotic strain.     

Detection of Salmonella spp. in stray dogs in Bursa Province, Turkey: first isolation of Salmonella Corvallis from dogs

The prevalence and antimicrobial sensitivity of Salmonella spp. in stray dogs in Bursa province, Turkey were determined from a total of 82 faecal samples by bacteriological methods. Of the dogs, nine (11%) were positive for salmonellae-carrying. All Salmonella isolates were serotyped as S. Corvallis, and were sensitive to amoxycillin/clavulanic acid, ampicillin, cephalothin, chloramphenicol, enrofloxacin, gentamicin, kanamycin, nalidixic acid, neomycin, oxytetracycline and trimethoprim/sulphamethoxazole, while 66.7% of them were resistant to streptomycin. It was concluded that stray dogs could be an important carrier of salmonellae as well as a source of human salmonellosis in Turkey. Additional studies are needed to clarify the epidemiological relationship between S. Corvallis isolated from dogs and humans with regard to public health. This is the first report on the isolation of S. Corvallis from dogs in the country.     

Characterization of four Flavobacterium columnare (Flexibacter columnaris) strains isolated from tropical fish

Four Flavobacterium columnare strains (AJS 1–4) were isolated from black mollies (Poecilia sphenops) and platies (Xiphophorus maculatus), showing white spots on the back, head and skin ulcers. The isolates developed characteristic rhizoid yellow pigmented colonies on Shieh agar and typical growth in Shieh broth. They were Gram-negative, filamentous bacteria exhibiting flexing movements. When compared to F. columnare strains isolated from temperate fish, it was noted that the four strains originating from tropical aquarium fish are more capable of growing at higher temperatures, the opposite being true for the strains isolated from temperate fish. Biochemical characterization and agglutination tests proved that the isolated strains could be classified as F. columnare. Low minimal inhibitory concentration (MIC) values were found for chloramphenicol, erythromycin, furazolidone, kanamycin, lincomycin, nalidixic acid, oxytetracycline and streptomycin. MIC values were high for colistin, sulfamethoxazole and neomycin. Pathogenicity studies were performed on black mollies. When these animals were submersed in an infective solution of the F. columnare strains, a marked difference in virulence was noted among the four isolated strains, strain AJS 1 being the most virulent one and strain AJS 4 being of low virulence.     

Prevalence of antimicrobial resistance among Salmonella on midwest and northeast USA dairy farms

The objective of this study was to determine the prevalence of antimicrobial resistance among Salmonella isolated from dairy herds in New York, Minnesota, Michigan, and Wisconsin, USA. Serogroup and antimicrobial susceptibility characteristics were determined for Salmonella from cattle and environmental samples collected during August 2000–October 2001 as part of a longitudinal study where 129 herds were visited at 2-month intervals. Salmonella isolates were tested (using a broth microdilution method) for susceptibility to amoxicillin/clavulanic acid, ampicillin, ceftiofur, ceftriaxone, cephalothin, chloramphenicol, ciprofloxacin, gentamicin, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, tetracycline, and trimethoprim/sulfamethoxazole. Of the 1506 isolates tested for minimum inhibitory concentrations to these 14 antimicrobial agents, 81.2% were pan-susceptible and for most herds (81.6%) the predominant antimicrobial resistance pattern was pan-susceptible. At least 1 Salmonella isolate resistant to 5 or more antimicrobial agents was found on 23.6% of herds. This resistance phenotype was most common among serogroups B and E1 and among samples from calves and farmer-designated sick cows. Resistant samples most frequently exhibited resistance to tetracycline, streptomycin, and/or ampicillin. No samples were resistant to ceftriaxone (though 13 were in the intermediate range), and very few samples were resistant to ciprofloxacin (n = 1), nalidixic acid (n = 5), or trimethoprim/sulfamethoxazole (n = 7).     

In vitro sensitivity of Hungarian Actinobaculum suis strains to selected antimicrobials

In vitro antimicrobial sensitivity of 12 Hungarian isolates and the type strain ATCC 33144 of Actinobaculum suis to different antimicrobial compounds was determined both by the agar dilution and by the disc diffusion method. By agar dilution, MIC50 values in the range of 0.05-3.125 micrograms/ml were determined for penicillin, ampicillin, ceftiofur, doxycycline, tylosin, pleuromutilins, chloramphenicol, florfenicol, enrofloxacin and lincomycin. The MIC50 value of oxytetracycline and spectinomycin was 6.25 and 12.5 micrograms/ml, respectively. For ofloxacin, flumequine, neomycin, streptomycin, gentamicin, nalidixic acid, nitrofurantoin and sulphamethoxazole + trimethoprim MIC50 values were in the range of 25-100 micrograms/ml. With the disc diffusion method, all strains were sensitive to penicillin, cephalosporins examined, chloramphenicol and florfenicol, tetracyclines examined, pleuromutilins, lincomycin and tylosin. Variable sensitivity was observed for fluoroquinolones (flumequine, enrofloxacin, ofloxacin), most of the strains were susceptible to marbofloxacin. Almost all strains were resistant to aminoglycosides but most of them were sensitive to spectinomycin. A strong correlation was determined for disc diffusion and MIC results (Spearman's rho 0.789, p < 0001). MIC values of the type strain and MIC50 values of other tested strains did not differ significantly. Few strains showed a partially distinct resistance pattern for erythromycin, lincomycin and ampicillin in both methods.     

In vitro comparison of the activity of various antibiotics and drugs against new Taiwan isolates and standard strains of avian mycoplasma

Twenty-nine antibiotics or drugs were incorporated individually into mycoplasma agar to evaluate their inhibitory activity against avian mycoplasmas: 100 recent Taiwan isolates of 7 serotypes and 10 standard strains of 7 serotypes were tested. All of the standard strains were very sensitive to erythromycin, chlorotetracycline, doxycycline, minocycline, and tetracycline, but the local isolates were highly resistant to these antibiotics. The drugs or antibiotics that possessed an MIC90 of 50 micrograms/ml or less against the local isolates were tiamulin (less than 0.4 micrograms/ml), lincospectin (2.7), josamycin (2.7), lincomycin (3.0), spectinomycin (4.8), tylosin (6.0), kanamycin (6.0), chloramphenicol (6.0), gentamicin (7.5), apramycin (24.5), doxycycline (27.4), minocycline (29.0), spiramycin (30.0), colistin (44.3), leucomycin (45.0), and streptomycin (50.0). The MIC90 of the other antibiotics or drugs was greater than 50 micrograms/ml. None of the isolates or strains were sensitive to nalidixic acid, ronidazole, penicillin, ampicillin, cephalexin, carbadox, or four sulfa drugs at a concentration about 5 times the therapeutic level.     

猪链球菌Ⅱ型四川分离株对抗菌药的敏感性分析

对四川省不同地区链球菌病病死猪体内分离的7株链球菌Ⅱ型进行抗菌药敏感性试验,用链球菌兰氏D群C55914作为对照,比对菌为肺炎链球菌质控菌株ATCC49619.在32种抗菌药中,选择29种药物进行纸片法药敏试验.试验结果表明:7株链球菌耐药谱非常相似,对青霉素、氨苄西林、阿莫西林、奥格门丁、卡那霉素、红霉素、壮观霉素、氯霉素、氟苯尼考、头孢氨苄、头孢拉定、头孢他定、头孢呋辛、头孢曲松、环丙沙星、氧氟沙星、二氟沙星、沙拉沙星、达氟沙星、恩诺沙星、复方新诺明、万古霉素、亚安培南均敏感;对阿米卡星、新霉素、庆大霉素3株敏感,4株中敏;对四环素和多西环素2株中敏,5株耐药;对链霉素1株中敏,6株耐药.用21种药物进行最小抑菌浓度(MIC)测定,结果7株均对青霉素、氨苄西林、卡那霉素、红霉素、头孢他定、头孢呋辛、头孢曲松、头孢噻呋、环丙沙星、达氟沙星、氧氟沙星、二氟沙星、沙拉沙星、恩诺沙星、磺胺六甲氧嘧啶敏感,1株对新霉素和阿米卡星中敏,2株耐药;所有菌株对四环素、土霉素、多西环素、链霉素均耐药.两种方法总体结果一致.     

In vitro susceptibilities of canine urinary bacteria to selected antimicrobial agents

Antimicrobial susceptibility tests were conducted on bacteria that were isolated from urine specimens collected by antepubic cystocentesis from dogs with urinary tract infections. Antimicrobics to which greater than or equal to 90% of these urinary bacteria were susceptible in vitro included trimethoprim-sulfamethoxazole (TMP-SMZ), nitrofurantoin, cephalexin, nalidixic acid, and gentamicin for isolates of Escherichia coli; ampicillin, TMP-SMZ, cephalexin, nalidixic acid, and gentamicin for isolates of Proteus mirabilis; ampicillin chloramphenicol, TMP-SMZ, nitrofurantoin, cephalexin, kanamycin, and gentamicin for isolates of coagulase-positive staphylococci; cephalexin, nalidixic acid, and gentamicin for isolates of Klebsiella pneumoniae; ampicillin, TMP-SMZ, and gentamicin for isolates of Streptococcus faecalis, Str faecium, and Str zymogenes; ampicillin, chloramphenicol, TMP-SMZ, and gentamicin for isolates of Str viridans; and ampicillin, chloramphenicol, TMP-SMZ, nitrofurantoin, cephalexin, kanamycin, and gentamicin for isolates of Str canis. No antimicrobial agent tested was effective in vitro at the 90% level for isolates of Pseudomonas aeruginosa, but gentamicin was closest, at 89%.     

The influence of flumequine on the resistance of the coliform flora of chickens to the agent

Flumequine (1 g/l) was administered to healthy young chickens in the drinking water for 5 days. A transient increase in the incidence of resistance to flumequine (4 micrograms/ml) was recorded in faecal coliform isolates after administration. Cross-resistance with nalidixic acid was demonstrated, but there was no evidence of cross-resistance to ampicillin, chloramphenicol, kanamycin, streptomycin, sulphadiazide, tetracycline and trimethoprim.     

Isolation of multiple drug-resistant enteric bacteria from feces of wild Western Lowland Gorilla (Gorilla gorilla gorilla) in Gabon

Prevalence of drug-resistant bacteria in wildlife can reveal the actual level of anthropological burden on the wildlife. In this study, we isolated two multiple drug-resistant strains, GG6-2 and GG6-1-1, from 27 fresh feces of wild western lowland gorillas in Moukalaba-Doudou National Park, Gabon. Isolates were identified as Achromobacter xylosoxidans and Providencia sp., respectively. Minimum inhibitory concentrations of the following 12 drugs—ampicillin (ABPC), cefazolin (CEZ), cefotaxime (CTX), streptomycin (SM), gentamicin (GM), kanamycin (KM), tetracycline (TC), nalidixic acid (NA), ciprofloxacin (CPFX), colistin (CL), chloramphenicol (CP) and trimethoprim (TMP)—were determined. Isolate GG6-2 was resistant to all antimicrobials tested and highly resistant to CTX, SM, TC, NA and TMP. Isolate GG6-1-1 was resistant to ABPC, CEZ, TC, CL, CP and TMP.     

Antimicrobial effect of combinations of EDTA-Tris and amikacin or neomycin on the microorganisms associated with otitis externa in dogs

Combinations of EDTA-Tris and two aminoglycoside antibiotics (amikacin and neomycin) were tested for synergistic activities against the microorganisms associated with otitis externa in dogs and for the solutions' stability over time. Synergistic activity was observed when EDTA-Tris plus amikacin and EDTA-Tris plus neomycin were tested againstStaphylococcus intermedius, Proteus mirabilis, Pseudomonas aeruginosa, andEscherichia coli, but not againstCandida albicans. Stability studies over a 3-month period indicated that the test solutions were stable at room temperature and that their antimicrobial activity was maintained.Abbreviations CFU colony-forming units - EDTA (ethylenedinitrilo)tetraacetic acid, disodium salt - FIC fractional inhibitory concentration - MBC minimal bactericidal concentration - MIC minimal inhibitory concentration - Tris tris(hydroxymethyl)aminomethane     

Resistance phenotypes and genotypes among multiple-antimicrobial-resistant Salmonella enterica subspecies enterica serovar Choleraesuis strains isolated between 2008 and 2012 from slaughter pigs in Okinawa Prefecture,Japan

A total of 349 Salmonella enterica subspecies enterica serovar Choleraesuis (S. Choleraesuis) strains, which were isolated between 2008 and 2012 from 349 pigs at two slaughterhouses in Okinawa Prefecture, Japan, were investigated for antimicrobial susceptibility and the presence of antimicrobial resistance genes. All isolates were resistant to at least four antimicrobial agents. The antimicrobial agents for which isolates showed a high incidence of resistance were as follows: ampicillin (100%) and streptomycin (100%), followed by gentamicin (99.7%), oxytetracycline (99.7%), sulfamethoxazole/trimethoprim (99.4%), nalidixic acid (40.1%) and oxolinic acid (40.1%). All isolates were sensitive to cefuroxime, ceftiofur, colistin, fosfomycin, enrofloxacin, orbifloxacin and danofloxacin. The predominant resistance phenotypes and genotypes were: resistance to ampicillin, streptomycin, gentamicin, oxytetracycline and sulfamethoxazole/trimethoprim (58.5%, 204/349) and blaTEM-strA-strB-aadA1-aadA2-aacC2-tet (B)-sul1-sul2-dhfrXII-dhfrXIII (36.1%, 126/349). The quinolone resistance-determining regions (QRDRs) of gyrA, gyrB, parC and parE of the quinolone-resistant isolates (n=12) showed amino acid substitutions of Ser-83→Phe or Asp-87→Tyr in GyrA and Ser-107→Ala in ParC. To our knowledge, this is the first report on the molecular characterization of antimicrobial resistance among S. Choleraesuis strains in Japan.     

In vitro antibacterial susceptibility of Arcobacter butzleri isolated from different sources

The objective of this study was to determine the antibiotic susceptibility of Arcobacter spp. isolated from various sources. Seventy Arcobacter spp. isolates were tested for their susceptibility to 13 antimicrobial agents. Antimicrobial susceptibility testing was performed by using the agar disc diffusion method on Mueller-Hinton agar supplemented with 5% defibrinated sheep blood. The antibiotics tested included enrofloxacin, erythromycin, streptomycin, gentamycin, rifampin, tetracycline, ampicillin, trimethoprim/sulfamethoxazole, nalidixic acid, danofloxacin, amoxycillin-clavulonic acid, cefuroxime-sodium, neomycine. Although all the arcobacters tested were susceptible to gentamycin, resistance to three or more antibacterial agents (especially, trimethoprim/sulfamethoxazole, cefuroxime-sodium and rifampin) was observed. A. butzleri isolates were found to be resistant to amoxycillin+clavulonic acid, nalidixic acid and ampicillin, at the rate of 20%, 44.28% and 78.57% respectively. In conclusion, gentamycin, streptomycin and tetracycline may be suitable antibiotics for the treatment or control of disease caused by Arcobacter spp. in veterinary and human medicine.