A bacterial pathogenicity determinant associated with necrotizing enterocolitis

Predominant enterobacteria from infants with necrotizing enterocolitis (NEC) were examined for an unusual ability to ferment lactose. One such isolate, a Klebsiella pneumoniae strain, was partially induced for lactose operon expression in tryptone containing media, and was also pathogenic in a rabbit ileal loop model for NEC. A spontaneous segregant of this strain was no longer partially induced for lactose operon expression, and was no longer pathogenic in the rabbit model. The gene responsible for this phenotype was cloned. The resulting plasmid was shown to cause both partially induced lactose operon expression and pathogenicity when introduced into a laboratory K. pneumoniae strain. A K. pneumoniae mutant deficient in lactose repressor synthesis was also pathogenic in the rabbit model. These results and previous studies on the intraluminal biochemistry of infants with NEC support the hypothesis that an increased ability for lactose fermentation may be a bacterial pathogenic trait with respect to NEC.     

Multiple antibiotic resistance in Staphylococcus aureus and Staphylococcus epidermidis: plasmids in strains associated with nosocomial infection

The plasmid DNA profiles were compared to phenotypically-similar, antibiotic-resistant strains of Staphylococcus aureus and Staphylococcus epidermidis associated with nosocomial infections in a Melbourne hospital. Whereas resistance to gentamicin, tobramycin and kanamycin was encoded by one of 3 plasmids [pSK1, 18 megadalton (Md); pSK4, 22 Md; pSK9, 17 Md] in S. aureus, no similar plasmids were detected in S. epidermidis. Mediated exclusively by the chromosome in S. aureus, tetracycline resistance was encoded either by the chromosome or by a 2.8 Md plasmid in strains of S. epidermidis. The inability to detect common resistance plasmids in strains of S. aureus and S. epidermidis recovered from this outbreak is in contrast to recent observations with staphylococci from other geographic areas; nevertheless, on the basis of restriction endonuclease analyses of 3 Md chloramphenicol resistance plasmids, it is suggested that a common gene pool does exist within isolates of S. aureus and S. epidermidis from Melbourne hospitals.     

Interactions among sex, HIV infection, and fat redistribution

The interactions among sex, HIV infection, and body fat redistribution are uncertain. We retrospectively compared total, subcutaneous, and visceral adipose tissue (TAT, SAT, VAT) contents, as determined by whole body MRI, in 85 HIV-infected persons, including 48 HIV-positive persons with self-reported changes in body shape, and matched healthy controls. The effect of sex on regional fat contents differed among HIV-infected persons with and without self-reported changes in body shape. Women without changes had significantly less SAT and TAT than did controls, while men with changes had significantly less SAT and TAT than did controls. Higher contents of VAT were found in both men and women with self-reported changes in body shape.     

Ten cases of Actinobaculum schaalii infection: clinical relevance, bacterial identification, and antibiotic susceptibility

Nine of 10 strains of Actinobaculum schaalii caused urinary tract infections in predisposed individuals. Identification included 16S rRNA gene sequence analysis and use of the API Coryne and Rapid ID 32 A test systems. A. schaalii is easily overlooked due to its slow growth in ambient air and its resemblance to the normal bacterial flora on skin and mucosa.     

Epidemiology of invasive pneumococcal infection in Taiwan: antibiotic resistance,serogroup distribution,and ribotypes analyses

From July, 1998, to June, 1999, pneumococcal isolates from 288 patients with invasive disease in Taiwan were serogrouped and tested for their susceptibility to various antibiotics. Automated ribotyping was used to study their molecular epidemiology. The mortality rate among those > or = 65 years was higher than those 18 or 19-64 years (p < 0.001). The total incidence of infection was significantly higher during the cooler season than the warmer season (p = 0.017). Among strains isolated from children aged < or = 18 years, 76% were not susceptible to penicillin, a rate that was significantly higher (p < 0.001) than that for adults (45%), as was the susceptibility to azithromycin, erythromycin, and trimethoprim-sulfamethoxazole (p < 0.005). The most prevalent serogroup encountered in the invasive isolates was 23, followed by 6, 14, 19, and 3. Isolation of Streptococcus pneumoniae in cerebrospinal fluid was at high rate in children under 5 years (p = 0.00012). Molecular typing revealed a high degree of polymorphism among the isolates. Among serogroup 23 and 19 isolates, a high proportion had the same ribotypes, the Taiwan23F-15 and Taiwan19F-14 isolates, suggesting the circulation of a Taiwanese epidemic strain. In Taiwan, S. pneumoniae isolates should be tested for their resistance profile for children < or = 18 years old, as these are more likely to harbor high-level resistance. Control of pneumococcal infection with the 7-valent-conjugated vaccine should also be considered because it is estimated that it would cover nearly 90% of the serotypes among pediatric invasive disease.     

Survey of antibiotic resistance among enterococci in North Rhine-Westphalia, Germany

A surveillance study on antibiotic resistance of enterococcal isolates (n = 730) was carried out in North Rhine-Westphalia, Germany, in 1997. Resistance rates to ampicillin (7.4%), high-level gentamicin (15.0%), high-level streptomycin (27.9%), ciprofloxacin (37.9%), vancomycin (1.5%), and teicoplanin (1.5%) were determined. All vancomycin-resistant enterococci (VRE) carried the vanA gene. SmaI and ApaI macrorestriction patterns indicated an intra- and interhospital spread of VRE.     

Exposure to ertapenem is possibly associated with Pseudomonas aeruginosa antibiotic resistance

The role of antibiotic exposure in the evolution and emergence of resistance is challenging to assess. We used carbapenem-resistant Pseudomonas aeruginosa (PA) phenotypes to assess possible factors that are associated with the occurrence and prognosis of such a phenotype and to examine the possible contribution of antibiotic exposure to the evolution of antimicrobial resistance. We conducted a nested case-control study. Cases were defined as patients from whom carbapenem-resistant ureidopenicillin-sensitive PA (CRUS-PA) was isolated; matched controls were PA patients who did not have isolation of CRUS-PA. We analysed potential predictors of CRUS-PA isolation and assessed their clinical significance (mortality and eventual isolation of pan-resistant PA), taking into account antibiotic exposures. We matched 800 case-control pairs. Case patients were more likely to have been exposed to anti-PA carbapenems (OR = 6.9; 95% CI, 2.5–18.6). This finding did not apply to the administration of other antibiotics. The mortality among CRUS-PA patients was similar to that of the controls (HR, 0.8 95%; CI, 0.6–1.1). Subsequent isolation of pan-resistant PA was more frequent among case patients compared with non-pan-resistant controls (p-value <0.05). Among cases, the risk of eventual pan-resistant PA isolation was increased in ertapenem recipients, only after and not prior to the index specimen date (HR, 1.9, 95%; CI, 1.01–3.4). Therefore we suggest that the CRUS-PA phenotype may represent pan beta-lactam resistance and that antibiotic exposure is associated with evolution of PA resistance phenotypes. We demonstrate a novel association of ertapenem with sequentially appearing PA resistance patterns.     

Synthesis, characterization, and in vitro activity of antibiotic releasing polyurethanes to prevent bacterial resistance

Central venous catheters are a major cause of nosocomial bloodstream infections. Different attempts have been made to incorporate antimicrobial agents into catheters, particularly directed at the surface-coating of devices. To facilitate the antimicrobial adsorption, various cationic surfactants, which however showed several problems, have been used. On the other hand, impregnated catheters with only antimicrobials have demonstrated a short-term duration due to the difficulties to deliver the drug slowly. Thus, in order to obtain high antimicrobial-polymer affinity we synthesized or modified polyurethanes to introduce different functional groups. Polymers were loaded with two antibiotics, cefamandole nafate and rifampin (RIF), chosen for both their functional groups and their action spectrum. The in vitro release behavior showed that the elution of drugs depended on the matrix hydrophilicity and on the antibiotic-polymer and antibiotic-antibiotic interactions. To increase the amount of drug released, polyethylene glycol (PEG) used as a pore forming agent at different molecular weights was incorporated in the polymer bulk with antibiotics. As for the in vitro antimicrobial activity of matrices, assessed by Kirby-Bauer test, it was seen that antibiotics released from various formulations inhibited the bacterial growth and exerted a synergistic effect when both were present. In particular, PEG10000-containing polymer was active against the RIF-resistant S. aureus strain up to 23 days. These results suggest that the combined entrapping of antibiotics and pore formers in these novel polymer systems could be promising to prevent the bacterial colonization and to control the emergence of bacterial resistance.     

Pseudomonas putida: identification, antibiotic sensitivity and pathogenicity (author's transl)]

This work studies 51 strains of Pseudomonas putida, isolated from clinical specimens (17) and hospital environment (34). Identification is performed by study of 41 physiologica and biochemical characters and 78 nutritional characters. According to the two biotypes A and B, described by Stanier, Palleroni and Doudoroff, these 51 strains can be grouped as follows: 48 have typical characters of biotype A, widely predominant, 3 can be distinguished from biotype A only by their auxanogram and included in biotype B. Antibiogram pattern of P. putida shows two salient features: resistant to carbenicillin and sensitivity to kanamycin. Among 17 human isolates, only 4 have likely pathogenic significance. By intraperitoneal challenge in mice, one half of strains is avirulent, other strains have a very low virulence (LD50: from 2,2 to 5 X 10(8) viable cells). There is no relationship between experimental virulence and bacterial sources.     

Vertebral osteomyelitis associated with single and mixed bacterial infection in broilers

Vertebral osteomyelitis (VO) is a worldwide emerging disease that affects broilers. The objective of this study was to determine the frequency and aetiology of VO in broilers in a highly productive broiler region. For this, 608 broilers with locomotory problems were analysed from 18 farms. Clinical signs were recorded, necropsy was performed and samples were collected from vertebral bodies with gross changes for molecular and histopathological analysis and for bacterial isolation. From broilers with locomotory changes, 5.1% (31/608) had VO and, of these, 93.5% were 40 days old or older and 89.7% were males. The birds with VO presented varying degrees of limited mobility and this was related to the level of compression to the spinal cord. Bacterial species of the genus Enterococcus (DNA detected in 53.6%) were the aetiological agents involved in most VO cases. Enterococcus faecalis was detected most frequently (35.7%), but Enterococcus hirae was also present in some lesions (7.1%). Escherichia coli was detected in 35.7% of vertebral lesions and co-infection with E. faecalis was confirmed in 7.1% cases. Staphylococcus aureus was involved in 14.3% of the cases, being 7.1% in co-infection with Enterococcus spp. or E. hirae. Our study has indicated that, in Brazil, VO in broilers may not be caused by a single infectious agent and has a lower frequency than recently reported in other countries. This study suggests that there are geographical differences between Brazil and other countries concerning the frequency and aetiology of VO.     

Serum resistance among Escherichia coli strains causing urinary tract infection in relation to O type and the carriage of hemolysin, colicin, and antibiotic resistance determinants.

The sensitivity to normal human serum of 91 smooth strains of Escherichia coli isolated from urinary tract infections was determined. Production of hemolysin, which was common and associated primarily with the types O4, O6, O18, and O75, was significantly correlated with high levels of serum resistance, both within the total population and within individual O types. In contrast, serum resistance was not significantly associated with antibiotic resistance (whether transmissible or not), with colicinogeny in general, or with colicin V production in particular. This indicates that the carriage of R and ColV plasmids, shown previously to be capable of conferring increased levels of serum resistance on individual strains of E. coli isolated from other sources, does not play an important part in determining the serum sensitivity of the E. coli population involved in urinary tract infection.     

Involvement of hepatic metallothioneins in hypozincemia associated with bacterial infection

Hypozincemia was induced in rats by Salmonella typhimurium and live vaccine strain Francisella tularensis (LVS) infections. Hepatic synthesis of zinc-binding proteins (ZBP) was studied in order to elucidate the mechanisms involved in the redistribution of zinc from plasma to liver occurring during infectious illness. ZBP, labeled in vivo with 65Zn, were isolated and identified as metallothioneins based, in part, on their heat stability, dimorphism, and amino acid composition. Cysteine was the major amino acid found in both forms of metallothionein and constituted 28-31% of total residues. The apparent half-life of these proteins as measured by disappearance of 65Zn was determined to be 19 h in a relatively mild infection (LVS) and 38 h in a more severe S. typhimurium infection. Results provide evidence that metallothioneins not only have the previously postulated regulatory role in normal zinc homeostasis but are intimately involved in the zinc redistribution occurring during the acute stage of infectious illness.     

Stimulation of mouse resistance to bacterial infection with muramyl dipeptide glycoside

We studied the capacity of 9 new muramyl dipeptide glycosides to stimulate mouse resistance to experimental sepsis induced by intraperitoneal injection of salmonella typhimurium culture. Preventive intraperitoneal injections of muramyl dipeptide -glycosides better improved survival of infected animals compared to the original (unmodified) muramyl dipeptide and muramyl dipeptide -glycosides. The most effective drug muramyl dipeptide -heptylglycoside injected during sepsis development also reduced animal mortality, decreased bacterial contamination of the viscera, and increased phagocytic activity of peritoneal macrophages in infected animals.     

Bartonella infection in animals: carriership, reservoir potential, pathogenicity, and zoonotic potential for human infection

Recent observations have begun to support a role for Bartonella spp. as animal as well as human pathogens. Bartonella spp. are vector-transmitted, blood-borne, intracellular, gram-negative bacteria that can induce prolonged infection in the host. Persistent infections in domestic and wild animals result in a substantial reservoir of Bartonella organisms in nature that can serve as a source for inadvertent human infection. The prevalence of bacteremia can range from 50 to 95% in selected rodent, cat, deer, and cattle populations. Dogs infected with Bartonella spp. can develop lameness, endocarditis, granulomatous lymphadenitis, and peliosis hepatis, lesions that have also been reported in association with human infection. Understanding the role of Bartonella spp. as pathogens in cats and other wild or domestic animals awaits the results of additional studies. Considering the extensive animal reservoirs and the large number of insects that have been implicated in the transmission of Bartonella spp., both animal and human exposure to these organisms may be more substantial than is currently believed.     

Synergistic effect of dosage and bacterial inoculum in TEM-1 mediated antibiotic resistance

The effect of inoculum size and gene dosage on the level of antibiotic resistance mediated by TEM-1-lactamase was measured. From the results it seemed that gene dosage is a more efficient mechanism than inoculum size for increasing TEM-1 mediated resistance to-lactam antibiotics. It also seemed that the two mechanisms for enhancing antibiotic resistance are synergistic. The clinical implications of these results are discussed.     

Distribution of Staphylococcus sciuri subspecies among human clinical specimens, and profile of antibiotic resistance

The distribution of three subspecies comprising Staphylococcus sciuri was determined for a collection of 30 clinical isolates originating from Morocco, the United Kingdom, and France. The sources of these isolates were principally wounds, skin, and soft tissue infections. At the species level, the isolates were identified according to biochemical characteristics and at the subspecies level by the ribotyping technique. PCR analysis performed with the 16S-23S ribosomal DNA intergenic spacer was less powerful for subspecies differentiation. S. sciuri subsp. sciuri was the most frequent subspecies (21 isolates) found in the collection, whereas S. sciuri subsp. rodentium (seven isolates) and S. sciuri subsp. carnaticus (two isolates) were less common. mecA or a mecA-related gene was detected by PCR and Southern blot in all 30 S. sciuri isolates, supporting the suggestion that S. sciuri species are the natural reservoir of the mecA gene. While the linA/linA' gene coding for lincomycin resistance was present in five isolates, an uncharacterized gene for this resistance was suspected in seventeen other isolates.     

A novel bacterial mucinase, glycosulfatase, is associated with bacterial vaginosis

The modifications to the vaginal habitat accompanying a change to vaginal flora in bacterial vaginosis (BV) are poorly understood. In this study enzymes involved in mucin degradation were measured, including a novel glycosulfatase assay. Women attending an emergency walk-in sexually transmitted disease clinic were studied. One high vaginal swab (HVS) was used to prepare a gram-stained smear to determine BV status, using Ison and Hay's criteria, and a separate swab was used for the purposes of the assays. The median glycosulfatase activity was 8.5 (range, -1.2 to 31.9) nmol h(-1) 1.5 ml(-1) of HVS suspension in patients with BV compared to 0.5 (range, -0.7 to 9.4) nmol h(-1) 1.5 ml(-1) of HVS suspension in patients without BV (P = <0.001). The median glycoprotein sialidase activity was 29.2 (range, -17 to 190) nmol h(-1) 1.5 ml(-1) of HVS suspension in patients with BV compared to -1.1 (range, -41 to 48) nmol h(-1) 1.5 ml(-1) of HVS suspension in patients without BV (P < 0.001). A rapid spot test for sialidase was positive in 22/24 patients with BV (sensitivity, 91.7%; 95% confidence interval [CI], 73 to 99%) and negative in 32/35 patients without BV (specificity, 91.4%; 95% CI, 76.9 to 98.2%) (P < 0.001). Glycosulfatase activity significantly correlated with both glycoprotein sialidase activity and the sialidase spot test (P = 0.006 and P < 0.001, respectively). The results are consistent with the hypothesis that the consortium of bacteria present in BV requires the ability to break down mucins in order to colonize the vagina and replace the normal lactobacilli.     

Mobile ISCR elements: Structure, functions, and role in emergence, increase, and spread of blocks of bacterial multiple antibiotic resistance genes

The recently discovered method of horizontal distribution of bacterial genes with ISCR atypical insertion sequences is reviewed using the example of drug-resistance genes. The mechanism of transposition of such elements including rolling circle replication, formation of autonomous nonreplicable circular structures, and homologous recombination provides mobilization of any section of the adjacent DNA. ISCR elements represent a more powerful gene mobilization system than transposons and integrons and provide formation of groups of mobile genes, including antibiotic resistance genes of pathogenic bacteria. The structure and functions of ISCR elements are discussed together with their similarity and dissimilarity to IS91-like elements and their role in emergence of blocks of multiple antibiotic resistance genes and their contribution to evolution of bacteria and plasmids.