ANTIBIOTIC-RESISTANT STAPHYLOCOCCUS INFECTIONS

Strains of antibiotic-resistant staphylococci that develop in a hospital are becoming increasingly important as a cause of untoward complications of hospitalization.The hospital environment and hospital personnel present the chief reservoir of antibiotic-resistant strains; and they, as well as patients discharged after hospital stay, may represent the greatest source of antibiotic-resistant strains found in the community at large.Cross infection appears to be thwarted by rigid adherence to antiseptic techniques.     

Transferable sodium arsenate resistance in enterobacteriaceae

The sodium arsenate and antibiotic resistance of a total of 750 strains of Enterobacteriaceae andPseudomonas aeruginosa isolated from hospital patients was determined. Resistance to sodium arsenate (minimal inhibitory concentration ≥25 mg/ml) was found in 61.4% of the isolates and was present both in antibiotic-susceptible and antibiotic-resistant strains. Ten enterobacterial isolates (3 sodium arsenate-resistant, antibiotic-susceptible strains and 7 sodium arsenate-resistant, antibiotic-resistant strains) could transfer sodium arsenate resistance to susceptible isolates and toEscherichia coli K-12 by conjugation. From sodium arsenate-and antibioticresistant strains, sodium arsenate and antibiotic resistances were transferred en bloc. Sodium arsenate resistance of antibiotic-susceptible strains was not self-transmissible, and transfer occurred only after R-plasmid mobilization.     

Estimation of the efficiency of ERIC-PCR typing of Mycobacterium tuberculosis for evaluation of the prevalence of antibiotic-resistant strains

Twenty-four antibiotic-resistant and sensitive strains of M. tuberculosis isolated from different territories of the Irkutsk region (East Siberia) were studied using PCR genotyping by enterobacterial repetitive intergeneric consensus (ERIC). Evolution relationships are illustrated by phylogenetic trees as a result of analysis by UPGMA and ML approaches. It was found that the studied samples belonged to two genetically different groups, both of which included sensitive and resistant strains. The sensitivity of the method was calculated by the Hunter-Gaston index. Based on these data, a probable pattern of emergence and propagation of antibiotic-resistant forms of tuberculosis in the studied region is discussed.     

Chromosomally integrated conjugative plasmids are common in antibiotic-resistant Haemophilus influenzae.

Twenty-three highly antibiotic-resistant strains of Haemophilus influenzae and two of Haemophilus parainfluenzae without detectable large plasmids were examined for conjugative transfer of their resistance to H. influenzae strain Rd or to other strains. Very inefficient transfer was observed for 18 H. influenzae strains and 1 H. parainfluenzae strain. All H. influenzae transcipients carried a large plasmid, and they were in turn efficient donors of their resistances in standard conjugation crosses with isogenic recipients. This was not seen for the H. parainfluenzae transcipients. It is concluded that most of the original antibiotic-resistant cultures carried an integrated conjugative R plasmid which had been excised in a few cells in each population. It was these cells which transferred resistance in the primary crosses.     

Plasmid transformation in Haemophilus influenzae.

Purified 34-megadalton-plasmid deoxyribonucleic acid from antibiotic-resistant strains of Haemophilus influenzae transforms competent strains of H. influenzae more efficiently if the recipient strains contain certain other 30-megadalton plasmids.     

Isolation of nonlysogenic bacteria from lambda-lysogenized strains using antibiotic-resistant phage

A simple and efficient method for obtaining nonlysogenic bacteria from lambda-lysogenized strains using phage with an inserted antibiotic-resistant plasmid is described. When the lysogenic culture is infected with antibiotic-resistant phage, single non-lysogenic cells are lysogenized, isolated on selective medium, and cured of phage during incubation at 37 degrees C.     

Significance of the antibiotic resistance of the causative agents of salmonellosis in its epidemiology and the procedure for controlling the infection

Antibiotic resistance of a wide set of strains (1501) of different serovars (69) was studied and the nature of the resistance was determined. Virulent bacteriophages designed with regard to the biological properties of the isolates were considered as possible agents for the control of antibiotic-resistant microorganisms. It was shown that multiple resistance to antibacterial drugs was mainly characteristic of the serovar of S. typhimurium. In Gorky and its region, strains carrying R plasmids determining the resistance to tetracycline and chloramphenicol with a molecular weight of 58-64 Md predominated. The antibiotic-resistant strains were dangerous from the epidemiological point of view. The use of the bacteriophages is advisable for the treatment of patients, sanation of bacterial carriers or decontamination of disease sources and prophylactic phaging with regard to the epidemiological indications for preventing group diseases.     

Identification and use of actinomycetes for enhanced nodulation of soybean co-inoculated with Bradyrhizobium japonicum

The utilization of actinomycetes as potential soybean (Glycine max (L.)) co-inoculants was evaluated. Soil samples from Carbondale and Belleville, Ill., were used to inoculate pre-germinated soybean plants to determine antibiotic sensitivity in the native Bradyrhizobium japonicum population. Sensitivity was in the order kanamycin > tetracycline > oxytetracycline > rifampicin > neomycin. Antagonism by five actinomycete cultures toward seven test strains of B. japonicum was also assessed. The ranking average inhibition (across all seven B. japonicum strains) by these actino mycetes was Streptomyces kanamyceticus = Streptomyces coeruleoprunus > Streptomyces rimosus > Streptomyces sp. > Amy colatopsis mediterranei. Ten antibiotic combinations were used to isolate antibiotic-resistant mutants of B. japonicum I-110 and 3I1B-110 via successive cycles of mutation. Eighty-one antibiotic-resistant strains were isolated and tested for symbiotic competency; nine of which were selected for further characterization in a greenhouse pot study. Few differences in nodule number were caused by these treatments. Nodule occupancy varied from 0% to 18.3% when antibiotic-resistant strains of B. japonicum were used as the sole inoculants. However, when three mutant strains of B. japonicum were co-inoculated with S. kanamyceticus, significant increases in nodule occupancy (up to 55%) occurred. Increases in shoot nitrogen composition (27.1%-40.9%) were also caused by co-inoculation with S. kanamyceticus.     

Association of qacE and qacEDelta1 with multiple resistance to antibiotics and antiseptics in clinical isolates of Gram-negative bacteria

Clinical isolates of Enterobacter cloacae, Citrobacter freundii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia were tested for resistance to antibiotics and to the antiseptics benzalkonium chloride and cetyltrimethylammonium bromide. Furthermore, they were examined for the presence of the resistance genes qacE and qacEDelta1. qacEDelta1 was detected by PCR in 10% of all (n=103) and in 81% of multiply antibiotic-resistant strains (n=15). qacE was found in only one of 37 P. aeruginosa strains. The minimum inhibitory concentrations of benzalkonium chloride, cetyltrimethylammonium bromide, and ethidium bromide were not significantly different for qacEDelta1/qacE-positive or -negative strains. Our data indicate that multiply antibiotic-resistant Gram-negative bacteria are not necessarily more resistant to quaternary ammonium compounds than antibiotic-sensitive strains even though qacE or qacEDelta1 is present.     

Antibiotic resistance of Lactobacillus strains]

One hundred and thirty six Lactobacillus strains isolated from poultry and 23 Lactobacillus strains isolated from long-living persons were tested for their antibiotic sensitivity. Occurrence of some type determinants of resistance to aminoglycoside antibiotics and tetracyclines in the Lactobacillus strains resistant to these antibiotics was studied. The majority of the strains from the both collections were resistant to aminoglycosides (73 and 79 per cent, respectively). The isolates from the poultry were characterized by multiple resistance. The isolates from the long-living persons were most frequently resistant to one of two antibiotics. All the tested Lactobacillus strains isolated from the long-living persons were sensitive to tetracyclines. The species composition of the isolates was different. The antibiotic-resistant strains were detected in all the species involved in the study. By hybridization of Lactobacillus colonies with the probes containing various genes of the resistance it was shown that in 14 per cent of the antibiotic-resistant strains belonging to Lactobacillus the antibiotic resistance was controlled by the genes homologous to resistance genes widely distributed in gramnegative organisms. This indicated a possible wide exchange and heterologous expression of the antibiotic resistance determinants between microorganisms of various taxonomic groups.     

Comparative survival of antibiotic-resistant and -sensitive fecal indicator bacteria in estuarine water.

The survival of antibiotic-resistant and -sensitive strains of Escherichia coli, Enterococcus faecalis, Enterococcus faecium, Streptococcus equinus, and two environmental isolates, AP17 and AQ62, was examined in estuarine water. Each strain was rendered resistant to a combination of two antibiotics by serial passage in increasing concentrations of antibiotics. Cultures were incubated in filter-sterilized estuarine water for up to 7 days. Recovery was assessed by examining colony-forming ability on media with and without antibiotics. None of the antibiotic-resistant forms survived longer than its antibiotic-sensitive counterpart in estuarine water. Three of the resistant strains died off more rapidly than the antibiotic-sensitive wild type. Survival of the test bacteria in estuarine water was as follows: sensitive and resistant AQ62, resistant Escherichia coli less than sensitive Escherichia coli less than resistant AP17 less than resistant Enterococcus faecium less than sensitive AP17, sensitive and resistant S. equinus less than sensitive and resistant Enterococcus faecalis, sensitive Enterococcus faecium. The results supported the suggestion that fecal entercocci may serve as better indicators of fecal pollution than Escherichia coli in marine ecosystems. Moreover, the results indicated that the use of antibiotic-resistant mutants to follow the fate of bacteria in the environment is inappropriate without adequate studies to ensure that resistant and wild-type strains react similarly to environmental stressors.     

Presence of chromosomal elements resembling the composite structure Tn3701 in streptococci.

Tn3701, carried by Streptococcus pyogenes A454, is the first chromosomal composite element to be described; it contains in its central region Tn3703, a transposon similar to Tn916. A comparison by DNA-DNA hybridization of Tn3701 with omega(cat-tet) and Tn3951, carried by Streptococcus pneumoniae BM6001 and by Streptococcus agalactiae B109, respectively, revealed that the two latter structures are also Tn3701-like composite elements. The DNAs of 27 other antibiotic-resistant group A, B, C, and G streptococci and of S. pneumoniae BM4200 showed sequence homologies to Tn3701 (14 strains, including BM4200), to the regions of Tn3701 outside of Tn3703 (5 strains), and to Tn916 alone (8 strains). The DNAs of five strains did not detectably hybridize with any probe. The tetM gene was identified in most chromosomal genetic elements coding for tetracycline-minocycline resistance. Since Tn3701-like elements are widely disseminated among antibiotic-resistant streptococci (47% of the 34 strains studied), we propose that Tn3701 be considered the prototype of chromosomal composite elements.     

Use of impedance measurements to estimate numbers of antibiotic resistant Salmonella strains

Impedance detection times were compared with traditional plating methods for enumerating antibiotic-resistant strains of Salmonella stanley, Salm. thompson and Salm. infantis grown in laboratory medium and pork slurry. The correleation of log₁₀ counts of salmonellas with detection times was highly significant ( r = -0·96 for broth and r = -0·94 for slurries. The confidence limits (± og₁₀ 1·0 for broth and ± log₁₀ 1·65 for slurry) indicated that detection times could reliably be used as a rapid means of enumerating salmonellas when large numbers of counts of known strains are required for growth studies. Use of antibiotic-resistant strains also permitted their selective detection by impedance from the natural spoilage flora of pork slurry when the same antibiotics were incorporated in the detection medium.     

The Synergistic Antibacterial Activity of Silver Nanoparticles and T. polium Extracts

Biophysics - The emergence of antibiotic-resistant bacterial strains leads to the need to find new antibacterial agents. Due to their properties, silver nanoparticles and plant extracts are more...     

Antibiotic resistance of gram-negative enteric bacteria from pigs in three herds with different histories of antibiotic exposure

The antibiotic resistance patterns of gram-negative fecal bacteria from pigs in three herds with different histories of antibiotic exposure were examined. In general, smaller proportions of antibiotic-resistant or multiply resistant fecal isolates (P less than 0.05) were obtained from pigs in a herd not exposed to antimicrobial agents for 154 months than from pigs in a herd continuously exposed to antimicrobial agents at subtherapeutic doses or from pigs in a herd exposed only to therapeutic doses of antimicrobial agents. The proportions of antibiotic-resistant and multiply resistant strains were greater among isolates from pigs in the therapeutic herd than in the non-antibiotic-exposed herd (P less than 0.05). The proportion of antibiotic-resistant isolates in the non-lactose-fermenting population was greater than that in the lactose-fermenting population, regardless of herd. The results suggest that any form of antimicrobial exposure will increase the prevalence of antimicrobial resistance and multiple resistance of fecal bacteria.     

Antibiotic resistance of gram-negative enteric bacteria from pigs in three herds with different histories of antibiotic exposure.

The antibiotic resistance patterns of gram-negative fecal bacteria from pigs in three herds with different histories of antibiotic exposure were examined. In general, smaller proportions of antibiotic-resistant or multiply resistant fecal isolates (P less than 0.05) were obtained from pigs in a herd not exposed to antimicrobial agents for 154 months than from pigs in a herd continuously exposed to antimicrobial agents at subtherapeutic doses or from pigs in a herd exposed only to therapeutic doses of antimicrobial agents. The proportions of antibiotic-resistant and multiply resistant strains were greater among isolates from pigs in the therapeutic herd than in the non-antibiotic-exposed herd (P less than 0.05). The proportion of antibiotic-resistant isolates in the non-lactose-fermenting population was greater than that in the lactose-fermenting population, regardless of herd. The results suggest that any form of antimicrobial exposure will increase the prevalence of antimicrobial resistance and multiple resistance of fecal bacteria.     

The sensitivity to honey of Gram-positive cocci of clinical significance isolated from wounds

AIMS: To determine the sensitivity to honey of Gram-positive cocci of clinical significance in wounds and demonstrate that inhibition is not exclusively due to osmotic effects. METHODS AND RESULTS: Eighteen strains of methicillin-resistant Staphylococcus aureus and seven strains of vancomycin-sensitive enterococci were isolated from infected wounds and 20 strains of vancomycin-resistant enterococci were isolated from hospital environmental surfaces. Using an agar incorporation technique to determine the minimum inhibitory concentration (MIC), their sensitivity to two natural honeys of median levels of antibacterial activity was established and compared with an artificial honey solution. For all of the strains tested, the MIC values against manuka and pasture honey were below 10% (v/v), but concentrations of artificial honey at least three times higher were required to achieve equivalent inhibition in vitro. Comparison of the MIC values of antibiotic-sensitive strains with their respective antibiotic-resistant strains demonstrated no marked differences in their susceptibilities to honey. CONCLUSIONS: The inhibition of bacteria by honey is not exclusively due to osmolarity. For the Gram-positive cocci tested, antibiotic-sensitive and -resistant strains showed similar sensitivity to honey. SIGNIFICANCE AND IMPACT OF THE STUDY: A possible role for honey in the treatment of wounds colonized by antibiotic-resistant bacteria is indicated.     

Increased Staphylococcus-killing activity of an antimicrobial peptide,lactoferricin B,with minocycline and monoacylglycerol

This study aimed to find antibiotics or other compounds that could increase the antimicrobial activity of an antimicrobial peptide, lactoferricin B (LFcin B), against Staphylococcus aureus, including antibiotic-resistant strains. Among conventional antibiotics, minocycline increased the bactericidal activity of LFcin B against S. aureus, but methicillin, ceftizoxime, and sulfamethoxazole-trimethoprim did not have such an effect. The combination of minocycline and LFcin B had synergistic effects against three antibiotic-resistant strains of S. aureus, according to result of checkerboard analysis. Screening of 33 compounds, including acids and salts, alcohols, amino acids, proteins and peptides, sugar, and lipids, showed that medium-chain monoacylglycerols increased the bactericidal activity of LFcin B against three S. aureus strains. The short-term killing test in water and the killing curve test in growing cultures showed that a combination of LFcin B and monolaurin (a monoacylglycerol with a 12-carbon acyl chain) killed S. aureus more rapidly than either agent alone. These findings may be helpful in the application of antimicrobial peptides in medical or other situations.     

Morphological heterogeneity of Bordetella pertussis populations and clones (based on electron microscopy data)

Populations belonging to different serovars of B. pertussis museum strains and antibiotic-resistant clones obtained from them have been studied by electron microscopy. As a result, morphological heterogeneity and differences in the ultrastructure of the cells with respect to the cell-wall structure, the character of the cytoplasm, the size of the cells, cytoplasmic inclusions and intracellular links have been demonstrated and, proceeding from these data, two main morphological variants of the cells have been defined. The cells of the morphological variant characterized by the pliciform surface of the outer membrane and the pronounced periplasmic space prevail among the populations of the museum strains. The possibility of the isolation of antibiotic-resistant clones, differing in their morphological structure and functional properties from the initial population, has been shown. The morphological diversity of B. pertussis population is the necessary condition for the existence and development of microbial populations.     

Agrobacterium-mediated transformation of large-flowered purslane (Portulaca grandiflora H.).

Transformation of Portulaca grandiflora has been developed with Agrobacterium tumefaciens strains A281 and T1272. Transformation was assessed by the following criteria: selection of hormone independent callus, antibiotic-resistant callus, and transgenic antibiotic-resistant plants. In addition, DNA hybridization analysis demonstrated that the DNA from tumor lines contained sequences homologous to binary vector T-DNA of strain A281. Following transformation with strain T1272, segregation analysis of the progeny of transgenic plants showed that the transgene was inherited in a Mendelian manner. The kanamycin-resistant progeny tested contained the T-DNA sequence of the strain T1272.