A one-year survey of respiratory and urinary pathogens and their antimicrobial susceptibility

Human cytomegalovirus (CMV) infection is often associated with myelosuppression and acute inflammatory reaction in immunocompromised patients. We have previously documented that CMV exposure of bone marrow (BM) stromal cells reduces the capacity of these cells to support hematopoiesis because of a decreased production of colony-stimulating factors. This study examines the potential role of CMV on constitutive and lipopolysaccharide (LPS)-stimulated production of cytokines involved in inflammatory reaction, interleukin-6 (IL-6) and leukemia inhibitory factor (LIF) by BM stromal cells. The release of IL-6 was already detectable 2 hours post CMV-infection (2.5-fold increase in production) and the cumulative production of IL-6 after 5 days of infection was 23 +/- 1.2 ng/mL (ninefold increase in production). CMV was also able to induce a time-dependent production of LIF that was maximal 8 hours after CMV infection (2.5-fold increase in production). Concomitantly, there was no detectable release of granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage CSF (GM-CSF) by CMV-infected stromal cells. The similar IL-6 and LIF production in the presence of polymyxin B ruled out the possibility that this increase could be caused by contamination of the viral stock by endotoxin. In addition, ultraviolet-inactivated virus behaved similarly to live virus and caused the release of IL-6 and LIF. However, heat-inactivated CMV was unable to induce IL-6 and LIF secretion by BM stromal cells. The production of IL-6 and LIF was also evaluated after stimulation by LPS. After 5 days of CMV exposure, the LPS-stimulated production of IL-6 and LIF was significantly lower than uninfected controls. This LPS-induced release of cytokine production was found to be dependent of viral replication. The experiments have shown that CMV is a potent inducer of IL-6 and LIF with differential effect on constitutive and LPS-stimulated cytokine production by stromal cells; we suggest that CMV induction of IL-6 and LIF during the first hours of infection could play a role in CMV-induced inflammatory reaction. Moreover, our results show that human CMV can disturb the balanced cytokine network involved in the regulation of hematopoiesis.     

The epidemiology of genitourinary fistulae in Kumasi, Ghana, 1977-1992

The aim of the study was to determine the clinical epidemiology of genitourinary fistulae as seen at Komfo Anokye Teaching Hospital in Kumasi, Ghana. A retrospective study was carried out from the hospital records and operative reports of all patients with genitourinary fistulae seen at Komfo Anokye Teaching Hospital between January 1977 and December 1992. Patient age, parity, type of fistula and cause of fistula were abstracted from the medical records. There were 164 cases of genitourinary fistula managed during the study period. There were 150 fistulae due to obstetric causes (91, 5%), the vast majority of which were due to prolonged obstructed labor (121 cases, 73.8% of all fistulae), with a minority related to complications of lower-segment cesarean section (14 cases, 8.5% of all fistulae). In 5 cases (3.1%) patients developed a rectovaginal fistula owing to perineal tears and prolonged obstructed labor. During this time period there were 157,449 deliveries, giving an obstetric fistula rate of 1 fistula per 1000 deliveries. Obstetric fistulae were most common at the extremes of reproductive age and parity Fourteen additional fistulae (8.5% of all cases) were due to gynecologic causes, most commonly from surgical injury occurring at the time of abdominal hysterectomy for leiomyomata uteri (12 cases, 7.3% of all fistulae). It was concluded that in Kumasi, Ghana, obstetric trauma from prolonged obstructed labor is the most common cause of genitourinary fistula formation. Such fistulae occur in older multiparous women as well in young primigravidae. Obstructed labour can, and does, occur in women who have previously undergone uneventful vaginal delivery. Birth attendants should be aware of that fact. Prompt referral for obstetric intervention should be made in obstructed labor, irrespective of the age and parity of the patient.     

Moraxella catarrhalis: clinical significance, antimicrobial susceptibility and BRO beta-lactamases

Moraxella catarrhalis is an important pathogen of humans. It is a common cause of respiratory infections, particularly otitis media in children and lower respiratory tract infections in the elderly. Colonisation of the upper respiratory tract appears to be associated with infection in many cases, although this association is not well understood. Nosocomial transmission is being increasingly documented and the emergence of this organism as a cause of bacteremia is of concern. The widespread production of a beta-lactamase enzyme renders Moraxella catarrhalis resistant to the penicillins. Cephalosporins and beta-lactamase inhibitor combinations are effective for treatment of beta-lactamase producers, and the organism remains nearly universally susceptible to the macrolides, fluoroquinolones, tetracyclines and the combination of trimethoprim and sulfamethoxazole. Two major beta-lactamase forms, BRO-1 and BRO-2, have been described on the basis of their isoelectric focusing patterns. The BRO-1 enzyme is found in the majority of beta-lactamase-producing isolates and confers a higher level of resistance to strains than BRO-2. The BRO enzymes are membrane associated and their production appears to be mediated by chromosomal determinants which are transmissible by an unknown mechanism. The origin of these novel proteins is unknown.     

A Polish multicenter survey of antimicrobial susceptibility and prevalence of beta-lactamase production among bacterial pathogens isolated from hospitalized and ambulatory patients

The aim of the study was to establish the frequency of occurrence of bacterial pathogens with beta-lactamase activity, and pattern of resistance among aerobic and anaerobic strains isolated from: respiratory tract, urinary tract, skin and soft tissues (hospitalized patients) and throat swabs (ambulatory patients). The study was conducted in 1994 year in 6 bacteriological laboratories in four Polish towns (Warszawa, Kraków, Katowice, Gdańsk) according to the protocol. Sensitivity of bacteria was tested by the disc method on the Müeller-Hinton agar or chocolate agar according to NCCLS, activity of beta-lactamase was tested with nitrocephin. A total 2038 clinical strains--1869 aerobic and 169 anaerobic was well-defined and tested for susceptibility to ten antibiotics--amoxicilin, augmentin, ofloxacin, gentamycin, cefradin, erythromycin, cefuroxim, kotrimoxazol, cefalexin and cefaclor. Among the isolated aerobes Staphylococcus aureus (25.1%), E. coli (23.2%) and Haemophilus influenzae (14.0%) were most frequent, and in the group of anaerobes the most frequent were Bacteroides spp (40.8%) We have found 45.8% of all tested aerobic strains with beta-lactamase production, the highest proportion in pathogens isolated from respiratory tract--51.4%, 46.6% from urinary tract, and 48.4% from skin and soft tissues. Among the isolated anaerobic--68.8% of Bacteroides and 28.6% others produced beta-lactamase. Forty percentage of all strains were sensitive to amoxicilin, 70-90% of aerobic bacteria were sensitive to augmentin. Augmentin had a high activity against anaerobic bacteria too. Only a small proportion of the tested aerobic bacteria (12.2%) were resistant to ofloxacin, gentamycin showed a sufficient activity against tested strains (24.4% were resistant). The most frequent pathogen--Staphylococcus aureus was resistant to amoxicilin in 83.1% hospitalized patients, and in 73.9% in ambulatory patients.     

Detection of Streptococcus pneumoniae in clinical specimens, its antimicrobial susceptibility and serotypes

The purpose of our investigation was the detection of S. pneumoniae and the isolation frequency of PRSP/PISP from the patients of the area. 457 strains of S. pneumoniae were isolated from the clinical specimens which had been requested for cultivation from the individual general practitioners in the city. Screening of the PRSP/PISP in 457 strains was carried out with the oxacillin (MPIPC) disk method. The MIC (benezylpenicillin: PCG) was measured at the same time in 73 strains, and compared with the disk method. The results were as follows: 1) Regarding the detection rate of S. pneumoniae, nose discharge was the highest (11.4%) in all the specimens. 2) There was a greater number of isolation of S. pneumoniae in one year from November to March. Then it was scarce in August and September. This change in seasons was noted. 3) There were many 9-year olds and over-60 year olds in the age group of the patients in whom the S. pneumoniae was isolated. 4) All 30 strains where a 6 mm diameter was found when a MIC was compared with the MPIPC disk method were resistant (PRSP/PISP). All 20 strains with a > or = 20 mm diameter were sensitive (PSSP). But in the 23 strains which had a 7 mm -19 mm, 7 strains (30%) were PSSP, and 16 strains (70%) in the intermediate (PISP). 5) A diameter of 6 mm was found in 194 strains (42.5%) with the MPIPC disk method, among the 457 strains isolated from the clinical specimens. 141 strains (30.9%) were found with a diameter of 7 mm -19 mm and 122 strains (26.7%) with a diameter of > or = 20 mm. 6) More than 70% in PRSP/PISP were multiple antimicrobiotics resistant strains to PCG, EM and MINO. 7) The distribution of the serotype in PRSP/PISP was in the order of 19 type (41.3%), 23 type (21.7%), 6 type (13.0%). Moreover, it was in the order of 3 type (18.5%), 6 type (11.1%), 19 type (11.1%), and there were few 23 type (3.7%) for PSSP. The distribution of the serotype was different for PRSP/PISP and PSSP.     

Bacterial pathogens in infected puncture wounds in adults with diabetes

Double immunohistochemical staining with rhodamine- and fluorescein isothiocyanate (FITC)-conjugated antisera revealed the coexistence of substance P (SP) and neuropeptide Y (NPY), and SP and calcitonin gene-related peptide (CGRP) in most nerve fibers in the intervascular stroma of the carotid labyrinth of the bullfrog, Rana catesbeiana, although there were a few fibers which showed only SP- or NPY-immunoreactivity. Approximately one third of SP-immunoreactive fibers also showed coexistence with vasoactive intestinal polypeptide (VIP)-immunoreactivity, and a few fibers contained VIP without SP. The combination of the double immunofluorescence technique and alternate consecutive sections further demonstrated the possible coexistence of SP, VIP, NPY, and CGRP. This coexistence of four different peptides in the same nerve fibers was proved by the following two evident facts: 1) some SP fibers which demonstrated coexistence with NPY-immunoreactivity were assumed to be continuous with those showing VIP-immunoreactivity, and 2) almost all of the SP fibers showed coexistence with CGRP-immunoreactivity. By this reasoning, nearly one third of SP fibers may demonstrate coexistence with NPY-, VIP-, and CGRP-immunoreactivities. These multiple peptides might be involved in vascular regulatory function, which is a possible function of the amphibian carotid labyrinth.     

Bacterial endotoxins and their effects

The lipopolysaccharide endotoxin macromolecules are cell wall's components of the Gram negative bacteria. The endotoxins are produced by Gram negative bacteria of intestinal flora. If the endotoxins are translocated from the intestinal tract to the circulation or injected into bloodstream, they elicit (depending from the quantity of endotoxin), slight or serious effects (e.g. endotoxin shock). In the effects of endotoxin certain cell populations (e.g. thrombocytes, macrophages, leukocytes, etc.), certain organs and organ-systems (e.g. liver, spleen, bone marrow, endocrine and lymphoreticular systems etc.) are involved. Effects of endotoxin are produced by mediators (e.g. endotoxin binding proteins, cytokines, prostaglandins, prostacyclins, NO etc.). The endotoxin sensitivity of vertebrate organisms is dependent from the phylogenetical status of the species. Most sensitive species is the human. Generally accepted that endotoxin has an important role in the pathogenesis of septic shock. In other pathological processes (e.g. intestinal syndrome of radiation disease, Gram negative infections, various shock forms etc.) are supposed or proved the role of endotoxins. Lead acetate induced endotoxin hypersensitivity or LAL methods are good tools for demonstration of the role of endotoxin in the pathogenesis of various processes. For this reason, the experimental endotoxin shock is used a model of septic and other shocks.     

Arabidopsis enhanced disease susceptibility mutants exhibit enhanced susceptibility to several bacterial pathogens and alterations in PR-1 gene expression

To identify plant defense responses that limit pathogen attack, Arabidopsis eds mutants that exhibit enhanced disease susceptibility to the virulent bacterial pathogen Pseudomonas syringae pv maculicola ES4326 were previously identified. In this study, we show that each of four eds mutants (eds5-1, eds6-1, eds7-1, and eds9-1) has a distinguishable phenotype with respect to the degree of susceptibility to a panel of bacterial phytopathogens and the ability to activate pathogenesis-related PR-1 gene expression after pathogen attack. None of the four eds mutants exhibited observable defects in mounting a hypersensitive response. Although all four eds mutants were also capable of mounting a systemic acquired resistance response, enhanced growth of P. s. maculicola ES4326 was still apparent in the secondarily infected leaves of three of the eds mutants. These data indicate that eds genes define a diverse set of previously unknown defense responses that affect resistance to virulent pathogens.     

In vitro antimicrobial susceptibility of Porphyromonas gingivalis to azithromycin, a novel macrolide

The in vitro antimicrobial susceptibility of Porphyromonas gingivalis to azithromycin, a new macrolide antibiotic of a new class known as azalides, was investigated by the agar dilution method on Brucella agar. Eighty-two P. gingivalis strains, 79 recent oral isolates, 1 nonoral isolate and 2 reference strains were included in the study. Azithromycin was highly effective against P. gingivalis. All strains were inhibited at 1.0 microgram/ml of azithromycin or less. The minimal inhibitory concentrations were 0.25 microgram/ml for 50% and 0.5 microgram/ml for 90%. These in vitro data as well as the favorable pharmacokinetics of azithromycin indicate that this new oral macrolide might be a good candidate for future clinical trials aiming to eradicate P. gingivalis from refractory periodontitis.     

Dermatophytes on the feet of HIV-infected patients: frequency, species distribution, localization and antimicrobial susceptibility

Skin scrapings from the toe clefts, soles and nail plates of 138 HIV-infected patients at various stages were examined for the presence of dermatophytes using both microscopy and culture. Dermatophytes, in particular Trichophyton rubrum, could be grown in 58 cases (42%). Although cultures were more often positive in late stages of disease, there was no close correlation with the clinical stage or the T4/T8 ratio. Susceptibility to itraconazole, but not to other antimycotics, was correlated with the immune status (P < 0.05). Pedal dermatophyte infection does not seem to be a major problem in HIV infection.     

Vitek system antimicrobial susceptibility testing of O1, O139, and non-O1 Vibrio cholerae

Vibrio cholerae causes epidemic diarrhea throughout the world. Fluid replacement is the primary therapy for cholera; however, high mortality rates often necessitate the use of antibiotics. V. cholerae, like most bacteria, has developed resistance to some antibiotics. In the early 1990s a new serotype strain, Bengal 0139, began a new wave of cholera epidemics. Bengal isolates showed unique trends in antimicrobial resistance. Many clinical laboratories use automated antibiotic susceptibility testing for V. cholerae. It is important to know if automated susceptibility test results for V. cholerae coincide with reported trends in antibiotic susceptibility. In the present study, we used the Vitek automated susceptibility system to determine the susceptibilities of 79 V. cholerae O1 isolates, 100 O139 isolates, and 112 non-O1 isolates. Vitek susceptibilities for V. cholerae showed a good correlation with preestablished epidemiological data. Although the new O139 serogroup showed a trend of increased resistance to trimethoprim-sulfamethoxazole and nitrofurantoin, it was more susceptible to ampicillin than previous serogroup O1 and non-O1 strains. Regardless of serogroup, > or = 98% of the V. cholerae isolates tested were susceptible to most antibiotics tested by us. It is important to continue susceptibility testing of all new isolates of V. cholerae because of emerging resistant strains. However, V. cholerae remains susceptible to most of the available antibiotics.     

Behaviour of hexahydrobenzodipyrazolones towards chloroacetylation, aminoalkylation, Grignard reagent and their antimicrobial activity

Treatment of 2,3a,4,6,7a,8-hexahydrobenzo [1,2-c; 4,5-c] dipyrazole-3,7-dione (1) with chloroacetyl chloride gave the 2,6-bis (chloroacetyl) derivative (2), which on treatment with acetic anhydride pyridine afforded (3). Compound (2) when heated with pyridine afforded (1). Compound (1) underwent Mannich reaction with piperidine or morpholine and formaldehyde to give the 2,6-bis (piperidino or morpholinomethyl) derivatives (4a,b). Hydroxymethylation of (1) with formaldehyde gave the 2,6-bis (hydroxylmethyl) derivative (4), which on heating with piperidine afforded (4a), Reaction of 2,3a,4,6,7a,8-hexahydro- 2,6-bis (phenylsulphonyl) benzo [1,2-c; 4,5-?] dipyrazole-3,7-dione (7) with phenylmagnesium bromide gave dodecahydro-3,3,4a,7,7,8a-hexaphenyl-2,6- bis (phenylsulphonyl) benzo [1,2-c; 4,5-?] dipyrazole (8). Derivatives of hexahydrobenzodipyrazolone (9a-g) have been subjected to general screening for their antimicrobial activity.     

Susceptibility pattern of Neisseria gonorrhoeae isolated at the Komfo, Anokye Teaching Hospital, Ghana to commonly prescribed antimicrobial agents

This study which aimed to determine the susceptibility patterns of seven often prescribed antimicrobial agents for treatment of gonorrhoea in Kumasi, Ghana, over a study period of 3 years (1991 through 1993), examined a total of 1187 specimens (high vaginal, cervical and urethral). Identification of N. gonorrhoeae was by colonial morphology, oxidase test, sugar fermentation test and the Gram-stain. The Intra-lactam strip method was used to determine which Neisseria gonorrhoeae isolates were beta lactamase producers (PPNG). Of the 1115 PPNG strains encountered, 300 were selected for MIC determination against seven (penicillin, cotrimoxazole, tetracycline erythromycin, spectinomycin, norfloxacin and ceftriaxone) antimicrobial agents in current usage in Kumasi, Ghana. Based on our results it was concluded, that PPNG strains are rampant/endemic in Kumasi, Ghana and that most exhibited multiple antimicrobial resistance. This situation requires, therefore, a serious review of the current Ghana Ministry of Health gonorrhoea treatment regimen. Suggestions to this end have been made.     

Serological characterisation and antimicrobial susceptibility of Actinobacillus pleuropneumoniae strains isolated from pigs in Spain

Seventy-one isolates of Actinobacillus pleuropneumoniae isolated from the lungs of pigs in outbreaks of pleuropneumonia in Spain were serotyped by indirect haemagglutination. Serotype 4 (42.2 per cent), serotype 7 (22.5 per cent) and serotype 2 (12.8 per cent) were predominant, whereas serotypes 1, 3, 6, 8, 9, 12 and untypable isolates were present only in small numbers. Serotypes 1, 2, 4 and 7 originated mainly from cases of acute pleuropneumonia, whereas serotypes 3, 6, 8, 9 and 12 were associated with chronically infected herds. The susceptibility of the isolates to 20 antimicrobial agents was determined by agar disc diffusion. Most were susceptible to cefuroxime, cefaclor, cefazolin, kanamycin, tobramycin, gentamicin, oxolinic acid, ciprofloxacin, enoxacin, thiamphenicol, colistin and trimethoprim/sulphamethoxazole. Marked resistance was found with amoxicillin, ticarcillin, oxytetracycline, doxycycline and metronidazole. Rifampicin, fosfomycin and tiamulin were the agents most effective against the isolates tested.     

Bacterial meningitis in emergency medicine. Factors associated with delay of antimicrobial therapy

Each year, Louisiana physicians are called upon to carry out mandatory preparticipation evaluations on more than 70,000 high school athletes and 3,000 college athletes. There have been changes in the guidelines for these evaluations in the state over the last few years. Also, recent state legislation has offered protection for the physicians providing them. This article outlines these changes and provides basic guidelines for the preparticipation evaluation.     

Two flavones from Artemisia giraldii and their antimicrobial activity

'Antiphospholipid' (aPL) antibodies are a heterogenous group of autoantibodies that are now considered to be directed mainly to plasma proteins or protein-phospholipid complexes. Standardisation of assays for anticardiolipin (aCL) antibodies and lupus anticoagulants (LA) have been fraught with difficulty despite numerous attempts to perform this by International Standardisation Workshops and Committees.