Susceptibility of Cheese and Yoghurt Starter Bacteria to Antibiotics

Eight single-strain lactic streptococci, three commercial cheese starters, and six lactic acid bacteria isolated from yoghurt were examined for their susceptibility to penicillin, cloxacillin, tetracycline-hydrochloride and streptomycin. The ranges of the antibiotics causing 50% inhibition of the bacteria were (mug/ml): penicillin, 0.009 to 0.20; cloxacillin, 0.24 to 2.50; tetracycline, 0.09 to 0.60; and streptomycin, 0.35 to 13.0. The average concentrations required to cause 50 and 100% inhibition of the cheese starters were (mug/ml): penicillin, 0.12 and 0.26; cloxacillin, 1.91 and 3.9; tetracycline-hydrochloride, 0.13 and 0.36; and streptomycin, 0.59 and 2.06. All the cocci were about equally susceptible to tetracycline, and all organisms were more resistant to cloxacillin than penicillin. The yoghurt isolates were more resistant to streptomycin and more susceptible to penicillin than the cheese starters. The 2, 3, 5-triphenyltetrazolium chloride test, using Streptococcus thermophilus BC as assay organism, does not detect low levels of streptomycin in milk. However, it is useful in detecting cloxacillin residues.     

Drug Resistance as Influenced by Inactivated Sensitivity Discs

Reports of staphylococci resistant to the semisynthetic penicillins stimulated a study of the factors influencing the stability of the drugs in discs. The behavior of penicillin G, methicillin, oxacillin, cloxacillin, and cephalothin discs under different humidity and temperature conditions is described. Humidity was found to be the most significant factor in drug inactivation. Storage of discs in a vacuum desiccator at -20 C provides maximal antibiotic stability.     

Antibiotic resistance and detection of β-lactamase in bacterial strains of Staphylococci and Escherichia coli isolated from foodstuffs

Seventy strains of Staphylococcus spp. and Escherichia coli (35 each) were isolated from various foodstuffs and identified on the basis of cultural, morphological and biochemical characteristics and were further tested for their antibiotic susceptibility with commonly used antibiotics/drugs. 69.2% of the strains of Staphylococci were resistant to co-trimazine and 34.6% were resistant to penicillin-G. 19.2% of the staphylococcal isolates exhibited resistance to cloxacillin, nalidixic acid, methicillin and tetracycline whereas 15.3% of the staphylococcal isolates were resistant to amoxycillin and nitrofurantoin. The isolated E. coli strains exhibited sharp peaks of resistance to antimicrobial agents such as tetracycline (72%), doxycycline (60%) and nalidixic acid (48%). Forty-four percent of the E. coli strains were resistant to nitrofurantoin and penicillin-G respectively. Among the 13 antibiotics/drugs tested for resistance, six different resistance patterns were observed in staphylococcal isolates and seven different resistance patterns were observed in the E. coli isolates from various foodstuffs. Bacterial strains exhibiting MIC values 100 g/ml for ampicillin and cloxacillin were screened for -lactamase activity and out of 10 staphylococcal isolates, seven were found to be positive for -lactamase, whereas out of 13 E. coli isolates tested for -lactamase production, only three were found to be positive.     

In Vitro and in Vivo Effects of Nitrofurantoin on Experimental Toxoplasmosis

Toxoplasma gondii is an important opportunistic pathogen that causes toxoplasmosis, which has very few therapeutic treatment options. The most effective therapy is a combination of pyrimethamine and sulfadiazine; however, their utility is limited because of drug toxicity and serious side effects. For these reasons, new drugs with lower toxicity are urgently needed. In this study, the compound, (Z)-1-[(5-nitrofuran-2-yl)methyleneamino]-imidazolidine-2,4-dione (nitrofurantoin), showed anti-T. gondii effects in vitro and in vivo. In HeLa cells, the selectivity of nitrofurantoin was 2.3, which was greater than that of pyrimethamine (0.9). In T. gondii-infected female ICR mice, the inhibition rate of T. gondii growth in the peritoneal cavity was 44.7% compared to the negative control group after 4-day treatment with 100 mg/kg of nitrofurantoin. In addition, hematology indicators showed that T. gondii infection-induced serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, biochemical parameters involved in liver injury, were reduced by nitrofurantoin significantly. Moreover, nitrofurantoin exerted significant effects on the index of antioxidant status, i.e., malondialdehyde (MDA) and glutathione (GSH). The nitrofurantoin-treated group inhibited the T. gondii-induced MDA levels while alleviating the decrease in GSH levels. Thus, nitrofurantoin is a potential anti-T. gondii candidate for clinical application.     

Inhibition of Aerobacter Cephalosporin β-Lactamase by Penicillins

Cephalosporinase (beta-lactamase) was obtained from cell washings of Aerobacter (Enterobacter) cloacae as a highly active preparation. An alkalimetric method was used to determine the enzyme activity and to estimate its inhibition by 6-amino-penicillanic acid derivatives. Their order of decreasing inhibitory effect was as follows: cloxacillin, oxacillin, methicillin, ampicillin, and penicillin G. We found that 2 to 3 ng of cloxacillin per ml was sufficient to decrease the enzyme activity by 50% in the presence of 400 mug of cephalosporin C per ml. Cloxacillin exerted a potentiating effect on the inhibition of the E. cloacae organisms by cephalosporin C.     

On-line clean-up and screening of oxacillin and cloxacillin in human urine and plasma with a weak ion exchange monolithic column

A weak ion exchange monolithic column prepared by modifying the GMA-MAA-EDMA (glycidyl methacrylate-methacrylic acid-ethylene glycol dimethacrylate) monoliths with ethylenediamine was applied to remove matrix compounds in biological fluid. Using this monolithic column, on-line clean-up and screening of oxacillin and cloxacillin in human urine and plasma samples had been investigated. Chromatography was performed by reversed-phase HPLC on a C(18) column with ultraviolet detection at 225 nm. Results showed that the ion exchange monolithic column could be used for deproteinization and retaining oxacillin and cloxacillin in human urine and plasma, which provided a simple and fast method for assaying drugs in human urine and plasma.     

Lack of benefit of ursodeoxycholic acid in drug-induced cholestasis in the rat.

The administration of ursodeoxycholic acid (UDCA) has been reported to improve cholestasis in patients with primary biliary cirrhosis or sclerosing cholangitis. In the present study, we tested the hypothesis that UDCA similarly might reduce cholestasis induced by drugs. Rats were treated with three different drugs reported to induce cholestasis: 17 alpha-ethynylestradiol, alpha-napthylisothiocyanate, and cyclosporine A. UDCA administration (0.4.g/day-1.k-1 before and during administration of the cholestatic drug) did not improve survival, food intake, or serum indicators of cholestasis in any of these three animal models of cholestasis. To the extent that drug-induced cholestasis in rats mimics the human situation, we conclude that UDCA probably will not be beneficial in drug-induced cholestasis in humans.     

Reversible renal failure in female patients with acute interstitial nephritis caused by cloxacillin]

Cloxacillin was not listed as one of the drugs causing the acute interstitial nephritis, yet. A case of a 50-year female patient treated with cloxacillin is presented. Therapy was followed by nausea, vomiting, myalgia and arthralgia, and the symptoms of the acute renal failure which completely diminished after prednisone therapy despite of co-existing peptic ulcer. Low doses of corticosteroids seem helpful in the treatment of the acute interstitial nephritis following therapy with cloxacillin even after a long time of the onset.     

NITROFURANTOIN (FURADANTIN?)—Use of Intravenous Forms in Resistant Surgical Infections; a Preliminary Report on 25 Patients

Twenty-five patients with severe and unusually resistant bacterial infections were treated with nitrofurantoin given intravenously.Twelve patients were classified as cured and seven as improved. In two cases there was no observable benefit. The other four patients, all moribund at the beginning of nitrofurantoin therapy, died. No significant toxic reaction to the drug was noted except for a tendency to metabolic acidosis in five patients in a state of shock after treatment with nitrofurantoin (Furadantin® intravenous solution). In no case was there evidence of impaired hematopoiesis.From this preliminary report it appeared that nitrofurantoin for intravenous use is justified in the treatment of gravely ill patients with surgical infections resistant to other antimicrobial drugs.     

Lithocholic acid decreases expression of bile salt export pump through farnesoid X receptor antagonist activity

Bile salt export pump (BSEP) is a major bile acid transporter in the liver. Mutations in BSEP result in progressive intrahepatic cholestasis, a severe liver disease that impairs bile flow and causes irreversible liver damage. BSEP is a target for inhibition and down-regulation by drugs and abnormal bile salt metabolites, and such inhibition and down-regulation may result in bile acid retention and intrahepatic cholestasis. In this study, we quantitatively analyzed the regulation of BSEP expression by FXR ligands in primary human hepatocytes and HepG2 cells. We demonstrate that BSEP expression is dramatically regulated by ligands of the nuclear receptor farnesoid X receptor (FXR). Both the endogenous FXR agonist chenodeoxycholate (CDCA) and synthetic FXR ligand GW4064 effectively increased BSEP mRNA in both cell types. This up-regulation was readily detectable at as early as 3 h, and the ligand potency for BSEP regulation correlates with the intrinsic activity on FXR. These results suggest BSEP as a direct target of FXR and support the recent report that the BSEP promoter is transactivated by FXR. In contrast to CDCA and GW4064, lithocholate (LCA), a hydrophobic bile acid and a potent inducer of cholestasis, strongly decreased BSEP expression. Previous studies did not identify LCA as an FXR antagonist ligand in cells, but we show here that LCA is an FXR antagonist with partial agonist activity in cells. In an in vitro co-activator association assay, LCA decreased CDCA- and GW4064-induced FXR activation with an IC(50) of 1 microm. In HepG2 cells, LCA also effectively antagonized GW4064-enhanced FXR transactivation. These data suggest that the toxic and cholestatic effect of LCA in animals may result from its down-regulation of BSEP through FXR. Taken together, these observations indicate that FXR plays an important role in BSEP gene expression and that FXR ligands may be potential therapeutic drugs for intrahepatic cholestasis.     

The cannabinoid agonist WIN 55, 212-2 increases nociception threshold in cholestatic rats: implications for the treatment of the pruritus of cholestasis

Dronabinol, a synthetic agonist at cannabinoid receptors, was reported to decrease the pruritus of cholestasis, in an uncontrolled observation. We hypothesized that the reported antipruritic effect of dronabinol might have resulted from an increased threshold to experience nociception (i.e. pruritus) by the drug. To test this hypothesis, we studied the effect of WIN 55, 212-2, a cannabinoid agonist, on the threshold to experience nociception, using a tail-flick assay in rats with cholestasis secondary to bile duct resection and in sham-resected controls. The administration of WIN 55, 212-2 was associated with a significant increase in the mean tail-flick latency in both groups as compared to baseline. Pruritus is a nociceptive stimulus; accordingly, drugs that increase the threshold to nociception in human beings may be a novel approach to the treatment of this symptom in patients with liver disease.     

Susceptibility of Genital Mycoplasmas to Antimicrobial Agents

The susceptibility of 11 T-strains, 12 strains of Mycoplasma hominis, and a single strain of M. fermentans to 15 antimicrobial agents was determined by study of inhibition of metabolic activity in a broth dilution system. All three species were inhibited by tetracycline, chloramphenicol, streptomycin, gentamicin, and kanamycin, and were relatively resistant to cephalothin, cephaloridine, polymyxin, vancomycin, and ampicillin. Three antimicrobial agents had significant differential effects on these species. Erythromycin was more active against T-strains than against M. hominis or M. fermentans. Lincomycin, clindamycin, and nitrofurantoin had greater activity against M. hominis and M. fermentans than against T-strains. The activity of the drugs tested was generally uniform over a wide range of inocula. The effect of pH and the difference between minimal inhibiting and minimal mycoplasmacidal concentrations of the drugs tested were consistent with expectations based on the effects of these drugs on bacteria.     

Generation of nitro radical anions of some 5-nitrofurans, 2- and 5-nitroimidazoles by norepinephrine, dopamine, and serotonin. A possible mechanism for neurotoxicity caused by nitroheterocyclic drugs

Catecholamine neurotransmitters such as norepinephrine, dopamine, and related catecholamine derivatives reduce nitroheterocyclic drugs such as nitrofurantoin, nifurtimox, nifuroxime, nitrofurazone, misonidazole, and metronidazole in slightly alkaline solutions. Drugs which contain 5-nitrofurans are reduced at lower pH than drugs which contain 2- and 5-nitroimidazoles. 5-Nitroimidazole derivatives such as metronidazole and ronidazole are known to be more difficult to reduce than 2-nitroimidazole derivatives, due to their lower redox potential. Catecholamines, when reducing nitro drugs, undergo concomitant oxidation to form semiquinone radicals. Both semiquinone radicals and nitro anion radicals formed in a reaction of nitro drug and catecholamine derivative were detected by electron spin resonance spectroscopy. Oxygen consumption studies in solutions containing nitro drug and catecholamine derivative showed that nitro anion radicals formed under aerobic conditions reduce oxygen to form the superoxide radical and hydrogen peroxide. Quinones formed in the reaction of catecholamine and nitro drug were detected by optical spectroscopy. Biosynthetic precursors and some metabolic products of catecholamines were also used in these studies, and they all exhibited reactions similar to catecholamines. Bovine chromaffin granules which synthesize and store catecholamines produced the nitrofurantoin anion radical when intact granules were treated with nitrofurantoin. These radicals formed inside the granules were observed by ESR spectroscopy. The formation of nitrofurantoin radical, semiquinone radicals of catecholamines, and oxygen-derived radicals by chromaffin granules is proposed to cause damage to adrenal medulla, and this process may lead to neurotoxicity.     

Studies on the induction of histidase in Vibrio el tor under nitrofurantoin resistant conditions

Studies on the induction of histidase were made with normal and nitrofurantoin resistant strains of Vibrio el tor. Nitrofurantoin resistant strains showed decreased level of induction in comparison to normal Vibrio el tor. The effect of different inhibitors like actinomycin D, chloramphenicol, tetracycline, nitrofurantoin and rifampicin on histidase induction was also studied. The mechanism of inhibition caused by the antibiotics is discussed.     

Reliability of the Kirby-Bauer Disc Diffusion Method for Detecting Methicillin-Resistant Strains of Staphylococcus aureus

The resistance of Staphylococcus aureus to methicillin and related drugs can be reliably determined by using the Kirby-Bauer method of susceptibility testing if the incubation temperature is 35 C or below, but resistance may be missed at 37 C. The 1-μg discs of oxacillin and nafcillin or the 5-μg discs of methicillin may be used for this purpose but not the 1-μg discs of cloxacillin. The latter fail to discriminate between sensitive and resistant staphylococci by zone measurement; some resistant strains of staphylococci may show larger zones of inhibition than sensitive strains. Stability of these antibiotic-containing discs was studied under conditions of temperature and humidity variation that might be encountered in a clinical laboratory refrigerator. Oxacillin discs were the most stable and are to be preferred for susceptibility testing. Nafcillin discs were less stable, and methicillin discs lose their potency rapidly unless carefully stored in a refrigerator with a desiccant.     

Antiarrhythmic drugs impair hepatic uptake and secretory function by different mechanisms in the isolated perfused rat liver

In the present study the effect of various antiarrhythmic drugs on hepatic perfusion parameters, uptake capacity of organic anions and biliary secretion using the isolated perfused rat liver was examined. Infusion of verapamil (VP), diltiazem, N-propyl-ajmaline (NPAB), and quinidine at pharmacological doses induced consistently a 1.4-1.6-fold increase in portal pressure accompanied by a approximately 60% decrease in bile flow and a approximately 65% inhibition of biliary taurocholate (TC) excretion. Furthermore, hepatic uptake of oxygen, bromosulphthalein (BSP), and TC was significantly reduced. All these effects were dose-dependent and reversible upon withdrawal of the drugs. Studies of the hepatic circulation using a Trypan blue staining technique demonstrated a patchy perfusion pattern during infusion of the antiarrhythmic drugs as compared to the homogenously stained control organ. The hemodynamic alterations and the impairment of the hepatic initial uptake function could be entirely prevented by concomitant administration of the vasodilator papaverine. Bile flow and biliary TC excretion, however, were still inhibited under these conditions. The present results indicate that antiarrhythmic drugs produce cholestasis in the isolated perfused rat liver independently of their adverse effect on hepatic hemodynamics.     

Prevalence of multiple antibiotic resistance among bacterial isolates from selected poultry waste dumps in Southwestern Nigeria

The prevalence of multiple antibiotic resistant bacteria in the waste dumpsite of ten poultry farms in Southwestern Nigeria was investigated. The susceptibility of 195 organisms isolated from the study sites to eight antimicrobial agents were tested using disc diffusion method and the minimum inhibitory concentration of cloxacillin and amoxicillin determined by the agar dilution method. Resistance to the test antibiotics ranged between 0% for gentamicin and 100% for tetracycline and ampicillin among the organisms. Overall, 70 and 90% of the isolates from Okuku, 65.2 and 95.6% from Ogbomoso, and 46.1 and 84.6% from Oyo had MIC above 512 μg/ml for amoxicillin and cloxacillin. Generally, drugs used in high volumes in the studied farms are the least active against the bacterial isolates. Results of this study shows that poultry waste can serve as environmental reservoirs of multiple antibiotic resistant bacteria and their indiscriminate dumping in the environment can expose surrounding human populations to health risks from drug resistant zoonotic pathogens. Part of the data presented in this paper was the subject of a presentation at the World Congress of Pharmacy and Pharmaceutical Sciences/67TH International Congress of the International Pharmaceutical Federation (FIP), 31 August—6 September 2007, Beijing, People’s Republic of China.     

Species differences in the interaction of serum albumin with antibiotics

Interaction of oxacillin, cloxacillin, dicloxacillin, phenoxymethylpenicillin, methicillin, nafcillin and benzylpenicillin with human serum albumin (HSA) was studied with flow microcalorimetry and differential scanning calorimetry. The measured thermodynamic parameters of complex formation between the penicillins and HSA were compared with similar characteristics of their binding to bovine serum albumin. It was shown that there were species differences between these two globular proteins in their interaction with the above antibiotics in relation to both the number of the biopolymer active sites and the nature of the molecular forces in the complex formation. The effect of the first bound molecule of oxacillin, cloxacillin, dicloxacillin, nafcillin, phenoxymethylpenicillin and benzylpenicillin on HSA conformation was observed. It was demonstrated that there was thermostabilization of HSA on its interaction with the above drugs with preserving cooperative nature of thermal denaturation of the complexes in relation to HSA melting.