Distribution Of Bacterial Species Research Articles

Genetic analysis of sea-ice bacterial communities of the Western Baltic Sea using an improved double gradient method

The bacterial diversity of sea ice from Kiel Bight obtained during the rare event of solid ice cover in spring 1996 was analysed by molecular genetic approaches using an improved double gradient denaturing gradient gel electrophoretic method (DG-DGGE) to separate 16S rDNA fragments of approximately 500 bp. The excellent separation of individual bands within these gradient gels allowed us to obtain sequence information and to allocate the phylogenetic position of representative bacteria from the sea ice. The band pattern of the gradient gels revealed a vertical stratification of the bacterial species distribution within the ice and the presence of characteristic bacteria for each layer. According to their 16S rDNA sequences, major bands of the gradient gels represented bacteria closely related to fermenting species of the genera Propionibacterium and Bacteroides and to anoxygenic phototrophic purple sulfur bacteria (Chromatiaceae). Their abundance in horizons of the inner ice core may indicate the existence of oxygen-deficient and anoxic zones or niches and possible primary production by anoxygenic photosynthesis within the investigated Baltic Sea sea ice. This is the first phylogenetic evidence of the presence, and most probably the development, of phototrophic purple sulfur bacteria in sea ice.

Changes in the Cultivable Flora in Deep Carious Lesions following a Stepwise Excavation Procedure

This study examined the cultivable microflora before and after stepwise excavation procedures in deep carious lesions in 9 permanent teeth, categorized according to degrees of proximal surface destruction. The final excavation was performed 4–6 months after the initial treatment, which included peripheral dentine excavation and removal of the central cariogenic biomass and the superficial necrotic dentine. Dentine colour and consistency were assessed by means of standardized scales before the application of a Ca(OH)₂ compound and temporary sealing. Reassessments were performed before and after the final excavation. Microbiological samples of the central demineralized dentine were obtained with a sterile bur before and after the first excavation, as well as before and after the final excavation. After anaerobic cultivation on enriched non–selective tryptic soy agar, 30 colonies from a representative area were identified by standardized biochemical and physiological tests. Before temporary restoration, a yellowish and light brown demineralized soft dentine was typically observed, and gram–positive rods accounted for 70% and lactobacilli for 50% of the total colony–forming units. Lactobacillus casei subsp. rhamnosus and Actinomyces naeslundii dominated the lactobacilli and the other gram–positive rods, respectively. Gram–negative rods were the next most frequent isolates, followed by streptococci, each group accounting for about 20% of the colony–forming units in positive samples. Before the final excavation, which did not cause exposure of the pulp in any of the cases, the retained demineralized dentine had changed into a darker and harder tissue, and the total colony–forming units, as well as the frequency and proportions of lactobacilli were substantially reduced. Gram–negative rods also declined, and the flora was dominated by A. naeslundii and various streptococci. In conclusion, the cultivable flora detected following the treatment interval had declined substantially, and the distribution of bacterial species did not represent a typical cariogenic microbiota of deep lesions, confirming the clinical findings of arrested caries progression.

Effect of soil type and plant species on the fluorescent pseudomonads nitrate dissimilating community

The distribution of nitrogen dissimilative abilities among 618 isolates of fluorescent pseudomonads was studied. These strains were isolated from two uncultivated soils (C and D; collected at Châteaurenard and Dijon, France, respectively) and from rhizosphere, rhizoplane and root tissue of two plant species (flax and tomato) cultivated on these two soils. According to their ability to dissimilate nitrogen, the isolates have been distributed into three metabolic types: non-dissimilators, NO2- accumulators and denitrifiers. While the three metabolic types were recovered in all the compartments of soil D experiments, only two (non-dissimilators and denitrifiers) were recovered in all the compartments of soil C experiments. Even under the contrasting conditions of the two soil types, both plants were able to select the nitrate dissimilating community among the total community of fluorescent Pseudomonas, but the mode of this selection seems to be dependent on both plant and soil type. The soil type appears to be unable to significantly modulate the strong selective effect of tomato. Indeed, similar dissimilator to non-dissimilator ratios were found in the root tissue of this plant species cultivated in both soils. In contrast, the different dissimilator to non-dissimilator ratios observed in flax roots between soils C and D suggest that the selective effect of flax was modulated by the soil type. Taxonomic identifications showed that the 618 isolates were distributed among three species (P. chlororaphis, P. fluorescens, P. putida) plus an intermediate type between P. fluorescens and P. putida. However, no clear relationship between the distribution of the metabolic types (functional diversity) and the distribution of bacterial species has been found.

A Multicentre Survey of Antimicrobial Resistance in Gram-Negative Isolates from Belgian Intensive Care Units in 1994-1995

SummaryThe aim of this prospective study was to evaluate the distribution and antibiotic susceptibility of aerobic Gram-negative bacilli isolated from patients in intensive care units in 18 Belgian hospitals during 1994 and 1995. A standardised method (i.e. the E-test) was used in each center to determine the minimum inhibitory concentrations of 12 major antibiotics against 1435 consecuti ve, non duplicate, Gram-negative isolates (close to 100 strains per hospital) during a period of 6 months. The isolates were mainly isolated from the lower respiratory tract (57.4%), urinary tract (17.7%), pus (7.9%) or blood specimens (7.8%) and were mainly P.aeruginosa (20.3%), E.coli (19.9%) and Enterobacter spp. (12.6%). Overall inducible Enterobacteriaceae (IE) accounted for 29.8% of all isolates, and E.aerogenes was the most frequently isolated species in this group (27.6%).The overall susceptibility rate (all species confounded) was about 70% to piperacillin, ticarcillin-clavulanic acid and ceftriaxone, 78% to piperacillin-tazobactam; 87% both to ceftazidime and to ciprofloxacin; and 90% to imipenem. Widespread resistance was observed in several IE species to third generation cephalosporins, broad-spectrum penicillins and to ciprofloxacin. By contrast, imipenem and the aminoglycosides still retained excellent activity against most multiresistant species.Although there were wide differences between hospitals in the frequencies of resistance to most antibiotics, these were not related to the types (general vs. university) of hospitals or to the number of beds. Some variations were however observed in the distribution of bacterial species: the prevalence of inducible Enterobacteriaceae was significantly higher in university than in general hospitals and in hospitals located in Brussels and in Wallonia than in the Flanders. Overall few trends in resistance rates were observed in comparison to a similar survey performed in 1991.

High frequency of antibiotic resistance among Gram-negative isolates in intensive care units at 10 Swedish hospitals.

OBJECTIVE: To investigate the resistance rates among Gram-negative isolates in Swedish intensive care units (ICUs). METHODS: During 1994-95, members of the Swedish Study Group collected, on clinical indication, 502 consecutive initial isolates of Gram-negative bacteria from patients admitted to ICUs at 10 Swedish hospitals and performed minimal inhibitory concentration (MIC) determinations with the Etest. Breakpoints were defined according to the criteria of the Swedish Reference Group for Antibiotics (SRGA). RESULTS: The distribution of bacterial species was: Escherichia coli > Klebsiella spp. > Enterobacter spp. > Pseudomonas aeruginosa > Haemophilus spp. > Proteus spp. > Stenotrophomonas maltophilia > Citrobacter spp. > Acinetobacter > Pseudomonas spp. > Morganella morganii > Serratia spp. Together these constituted 97% of all isolates. The frequencies of resistance for all the initial Gram-negative isolates were: ceftazidime 6.8%, cefotaxime 14.9%, ceftriaxone 18.5%, cefuroxime 44.1%, ciprofloxacin 4.2%, co-trimoxazole 17.8%, gentamicin 5.8%, imipenem 8.6%, piperacillin 20.2%, piperacillin/tazobactam 12.9% and tobramycin 5.8%. CONCLUSIONS: Among Gram-negative isolates in Swedish ICUs, a very high frequency of resistance was seen to cefuroxime, and rather high frequencies of resistance to cefotaxime, ceftriaxone, piperacillin and piperacillin/tazobactam. These drugs cannot be recommended for further use as empirical monotherapy for severe ICU-acquired Gram-negative infections in ICUs in Sweden.

Bacterial vaginosis in pregnancy: distribution of bacterial species in different gram-stain categories of the vaginal flora

Vaginal swabs for microbiological culture were taken from 174 pregnant women whose vaginal flora had been evaluated by Gram's stain; 50 had grade III flora (bacterial vaginosis, BV), 50 grade II (intermediate), 41 had vaginal flora graded as abnormal which then reverted to grade I (revertants) and 33 had normal flora (controls). The aim was to determine whether bacterial species isolated from women with grade II flora differed from those with grade III flora. Isolation of Lactobacillus spp. decreased from grade I to grade III and that of other aerobic and anaerobic bacterial species increased. There was little difference in the species isolated from women with grade II and grade III flora, but there was a distinct order in which organisms in different species increased in numbers. The vaginal flora of revertants was intermediate between that of healthy controls and those with grade II flora. Coagulase-negative Staphylococcus spp. were isolated from a greater number of revertants than grade I controls but the incidence did not increase in grade II or grade III. Bifidobacterium spp. were isolated from a greater number of revertants than grade I controls and increased further in grade II and grade III. However, Gardnerella vaginalis and Mycoplasma hominis were isolated from a much larger number of women with grade III flora than the other groups. The conclusion is that grade II is a transitional phase between grade I and grade III and that some organisms such as G. vaginalis and M. hominis only reach large numbers in the late stage. The sequence of appearance of the various bacterial species may be a result of the pathophysiological alteration of the vaginal ecosystem associated with BV.

Effect of oxytetracycline-medicated feed on antibiotic resistance of gram-negative bacteria in catfish ponds

The effect of oxytetracycline-medicated feeds on antibiotic resistance in gram-negative bacteria from fish intestines and water in catfish ponds was investigated. In experiments in the fall and spring, using ponds with no previous history of antibiotic usage, percentages of tetracycline-resistant bacteria in catfish intestines obtained from medicated ponds increased significantly after 10 days of treatment. In the fall, resistance of the intestinal and aquatic bacteria returned to pretreatment levels within 21 days after treatment. In the spring, resistance declined after treatment but remained higher than pretreatment levels for at least 21 days in intestinal bacteria and for 5 months in aquatic bacteria. Plesiomonas shigelloides, Aeromonas hydrophila, and Citrobacter freundii were isolated frequently in both spring and fall; Escherichia coli, Klebsiella pneumoniae, Edwardsiella tarda, and Enterobacter spp. were isolated primarily in the spring. Oxytetracycline treatment did not affect the distribution of bacterial species in the fall but may have accelerated a shift toward greater prevalence of members of the family Enterobacteriaceae in the spring. Multiple antibiotic resistance did not appear to be elicited by oxytetracycline treatment.

Resistance to cefotaxime and seven other beta-lactams in members of the family Enterobacteriaceae: a 3-year survey in France.

During the second quarter each of 1988, 1989, and 1990, a French collaborative study group, including 12 university hospital laboratories, surveyed the resistance to beta-lactams of clinical isolates from hospitalized patients: consecutively, 10,641, 10,692, and 9,382 isolates were tested. The distribution of bacterial species over time was similar in each laboratory. The susceptibilities of microorganisms to amoxicillin, ticarcillin, cephalothin, cefoxitin, cefotaxime (CTX), ceftazidime (CAZ), aztreonam (ATM), and imipenem (IPM) were measured by the disk diffusion method in accordance with the recommendations of the Antibiogram Committee of the French Society for Microbiology. Five reference strains were included for quality control. Extended-spectrum beta-lactamases were detected by the synergistic effect of the combination of clavulanic acid-amoxicillin with CTX, CAZ, and ATM in the double-diffusion test. A synergistic effect with CTX, CAZ, and ATM was detected for 1.5% of all strains, mainly those of Klebsiella pneumoniae (13.3%). For this species, the synergy test enabled the detection of roughly 50% of the resistant strains misclassified as susceptible on the basis of interpretative standards. Extended-spectrum beta-lactamases disseminated in 1990 in most enterobacterial species but at a low frequency. Important variations in the percentages of resistant strains were observed in terms of bacterial species, hospitals, and wards. However, when the total number of strains was considered, the percentages of resistance to newer beta-lactams remained low.

Bacterial Sampling by Absorbent Paper Points. An in vitro Study

This in vitro study was designed to test the hypothesis that sampling of bacterial suspensions by absorbent paper points would accurately reflect the proportion of the bacterial species whether they are homogeneously or non-homogeneously distributed in the sample site. The non-homogeneously distributed populations consisted of equal numbers of colony forming units (CFU) of two periodontal bacterial species present as separate cultures, one layered on top of the other. The species in the top layer accounted for greater than 90% of the total number of CFU detected by paper point sampling, regardless of which species formed the top layer. In contrast, when the layers were mixed prior to sampling to give a homogeneous bacterial distribution, the paper points detected equal numbers of CFU of each species. Thus the hypothesis was not supported and the proportions yielded by paper point sampling are not the same when the bacteria are non-homogeneously distributed as when they are homogeneously distributed. Ultrastructural observations have documented a non-homogeneous distribution of bacterial species in periodontal pockets. If the specific microbiota in the apical portion of deep periodontal pockets are important to the periodontal disease process, these data suggest that bacterial samples obtained by absorbent paper points misrepresent the presence of such bacterial species.

Should potted plants be removed from hospital wards?

Seventy-nine isolates of Gram-negative bacilli were recovered from 29 potted plants on six surgical wards. The distribution of bacterial species and antibiotic susceptibility patterns revealed no correlation with 235 isolates from nearby patients during the study period. Potted plants do not appear to constitute a bacteriological hazard in the hospital.

Impact of short perioperative courses of cefotaxime on aerobic bacterial flora in patients undergoing transurethral prostatic resection.

The influence of short-term prophylactic courses of cefotaxime on the microbial environment was studied. The distribution of bacterial species and their antibiotic resistance was recorded in isolates collected over a six-year period, during which time cefotaxime was used for perioperative prophylaxis in patients undergoing transurethral prostatic resection. In three consecutive studies covering the six-year observation period, the species distribution and antibiotic resistance patterns remained essentially unchanged for both cefotaxime and ampicillin. Examination of the faecal coliform flora in 23 patients given cefotaxime revealed no marked qualitative or quantitative change in the flora. It is concluded that short-term prophylactic courses of cefotaxime do not promote the emergence of resistance in the aerobic bacterial flora.

The Normal Cultivable Microflora in Upper Jejunal Fluid in Healthy Adults

Bacteriological studies of uncontaminated upper jejunal fluid were performed in 85 normal subjects. Fifty-three per cent of the samples were sterile (less than 10(1) CFU/ml). In 10% of the cases the total number of microorganisms exceeded 10(5) CFU/ml. The main groups of microorganisms isolated were Streptococcus sp ('Viridans group'), Lactobacillus sp., Veillonella parvula, Actinomyces sp., Haemophilus sp., Corynebacterium sp., and Candida albicans, each found in more than 10% of the subjects. Only the Streptococcus sp. exceeded 10(5) CFU/ml, and enterobacteria were found in 5% of the subjects, the number not exceeding 10(3) CFU/ml. No other typical members of the lower gastrointestinal tract were isolated. The number of subjects harbouring bacteria and the distribution of bacterial species were the same in both sexes and in different age groups.