Staphylococci in skin microbiocenosis of the breasts in nursing women

The levels of the bacterial contamination of the nipple, the areola and the surrounding skin, the occurrence and species composition of staphylococci in 120 nursing women on days 4-5 after parturition have been studied. S. aureus contaminate the surface of the nipple and the areola in 75% of the examined women, and in 57.5% of these women the massive contamination of the above-mentioned areas (greater than or equal to 10(3) colony-forming units per sq. cm) is observed. In 80% of puerperae the occurrence of S. epidermidis on the nipple, the areola and the surrounding skin has proved to be practically the same. The population of S. aureus colonizing the mammary glands consists mainly of hospital strains; of these, 75.97% belong to phage type 75.     

Staphylococci in skin microbiocenosis of the breasts in healthy women

The autoflora of different anatomical regions of the mammary glands in 120 healthy nulliparous women aged 18-24 years was studied by P. Williamson and A. M. Kligler's methods of smears and washings. From the nipple, the areola, and the adjacent region of the skin 2,248 strains of anaerobic microorganisms were isolated; of these, 63.83% were staphylococci and micrococci, 6.01% were streptococci, 1.91% were Neisseria, 17.79% were Corynebacterium, 3.87% were bacilli, 2.8% were enterobacteria, and 3.79% were fungi. Coagulase-positive staphylococci occurred in 1.56% of cases. Out of 11 coagulase-negative species of this genus, S. epidermidis occurred most frequently on the skin of the mammary glands. The nipple was found to have the highest bacterial contamination (0.55 X 10(6) +/- 0.7 X 10(5) cells/sq. cm for the right mamma and 0.59 X 10(6) +/- 0.7 X 10(5) cells/sq. cm for the left mamma) and the skin adjacent the areola, the lowest bacterial contamination (0.14 X 10(4) +/- 0.2 X 10(3) cells/sq. cm for the right mamma and 0.25 X 10(4) +/- 0.3 X 10(3) cells/sq. cm for the left mamma). P. Williams and A. M. Kligman's method of washings, more accurate and informative, was found to be preferable for the study of the quantitative characteristics of the dermal microbiocenosis of the mammary glands.     

Enumeration of mycoplasmas after acridine orange staining

Samples of liquid mycoplasma cultures were mixed with equal part of a 0.01% solution of acridine orange and placed on agar plates. The number of fluorescing organisms per field was counted in an epifluorescence microscope at an X 1,000 magnification. When the number of fluorescing organisms per field was related to the number of colony-forming units per milliliter during the growth cycle, highly significant correlation was found in cultures with greater than or equal to 10(6) colony-forming units per ml during the exponential growth phase. The counts were weakly correlated during the stationary phase and not correlated during the death phase. This technique provides a mean to enumerate mycoplasmas in liquid cultures.     

Enumeration of mycoplasmas after acridine orange staining.

Samples of liquid mycoplasma cultures were mixed with equal part of a 0.01% solution of acridine orange and placed on agar plates. The number of fluorescing organisms per field was counted in an epifluorescence microscope at an X 1,000 magnification. When the number of fluorescing organisms per field was related to the number of colony-forming units per milliliter during the growth cycle, highly significant correlation was found in cultures with greater than or equal to 10(6) colony-forming units per ml during the exponential growth phase. The counts were weakly correlated during the stationary phase and not correlated during the death phase. This technique provides a mean to enumerate mycoplasmas in liquid cultures.     

Genetic transformation of Streptococcus pneumoniae by heterologous plasmid deoxyribonucleic acid.

A number of heterologous plasmid deoxyribonucleic acids (DNAs) coding for erythromycin, tylosin, lincomycin, tetracycline, or chloramphenicol resistance have been introduced into Streptococcus pneumoniae via genetic transformation with frequencies that varied between 10(-5) to as high as 5 x 10(-1) per colony-forming unit. Transformation with plasmid DNA required pneumococcal competence, was competed by chromosomal DNA, and showed a saturation at about 0.5 micrograms/ml (with a recipient population of 3 x 10(7) colony-forming units of competent cells per ml). Plasmid transformation did not occur with a recipient strain, 410, defective in endonuclease I activity and in chromosomal genetic transformation. All erythromycin-resistant transformants examined contained covalently closed circular DNA with the same electrophoretic mobility on agarose gels as the donor DNAs, and when examined in detail the plasmid reisolated from the transformants had the same restriction patterns and the same specific transforming activity as the donor DNA. In the cases of two plasmids examined in detail--pAM77 and pSA5700 Lc9--most of the transforming activity was associated with DNA monomers; DNA multimers present in pSA5700 Lc9 also had biological activity. An unexpected finding was the demonstration of transformation (2 x 10(-5) per colony-forming unit) with plasmid DNAs linearized by treatment with S1 nuclease or with restriction endonucleases.     

Estimation of the Growth of the T1 Strain of Mycoplasma mycoides in Tryptose Broth by the Measurement of Lactate Dehydrogenase: I. Method

A highly significant correlation coefficient (r = 0.97, n = 18) was found between the concentration of lactate dehydrogenase measurable after the organisms had been disrupted and the concentration of colony-forming units during the logarithmic phase of growth of a broth culture of the T(1) strain of Mycoplasma mycoides var. mycoides. A concentration of 4.60 x 10(-7) milliunits of lactate dehydrogenase for each colony-forming unit was established. This relationship was used to convert the concentration of lactate dehydrogenase in the culture into an estimate of the concentration of viable mycoplasma. The lactate dehydrogenase was estimated by following the oxidation of reduced nicotinamide adenine dinucleotide, in the presence of pyruvate substrate, at 366 nm in a spectrophotometer. The nicotinamide adenine dinucleotide oxidase system probably contributed a small amount of enzyme activity to the test when lactate dehydrogenase was measured in this way. The method has been described and evaluated for the estimation of titers from 10(7) to 5 x 10(9) colony-forming units per ml.     

Adenylate energy charge in Acholeplasma laidlawii.

Adenosine 5'-triphosphate, adenosine 5'-diphosphate, and adenosine 5'-monophosphate were produced by Acholeplasma laidlawii B-PG9 growing in modified Edward medium. The adenylate energy charge was calculated to be 0.84 +/- 0.07 and ranged from 0.91 to 0.78 during exponential growth (12 to 24 h). During exponential growth, A. laidlawii contained, at 17.5 h, 2.3 X 10(-17) mol of adenosine 5'-triphosphate per colony-forming unit and, at 16 h, 27.3 nmol of adenosine 5'-triphosphate per mg (dry weight). The medium supported a doubling time of 0.95 h. The molar growth yields (Yglucose = grams [dry weight] per mole of glucose used) were 40.2 +/- 3.4 (16 h) and 57.1 +/- 9.7 (20 h) during midexponential growth. A maximum yield of 8.3 X 10(9) colony-forming units was reached at 24 h, when 56% of the initial concentration of glucose had been used. At 40 h, during the stationary phase, 14.95 +/- 3.75 mumol of glucose per ml of medium had been used. At this time, the culture fluids contained 21.86 +/0 mumol of lactate per ml and 3.14 +/- 0.13 mumol of pyruvate per ml.     

Mass Airflow Cabinet for Control of Airborne Infection of Laboratory Rodents

A mass airflow cabinet for handling and housing of laboratory rodents has been developed and tested. The unit consists of a high-efficiency particulate air filter and uniform distribution of air at a vertical velocity of 19 cm per s. Animals are maintained without bedding in mesh-bottomed cages that rest on rollers for rotation inside the cabinet. There is an air barrier of 90 cm per s separating the cabinet air from room air. Sampling for airborne bacteria yielded an average of 0.03 colony-forming units (CFU) per ft(3) of air inside the cabinet, whereas 28.8 CFU per ft(3) was simultaneously detected outside the cabinet during housekeeping, a reduction of almost three logs. The efficiency of the air barrier was tested by aerosolization of T3 phage. When phage was aerosolized 5 cm outside the cabinet, no phage could be detected 5 cm inside when the fans were operating; with the fans off an average of 1.6 x 10(4) plaque-forming units (PFU) per ft(3) was detected in six tests. Aerosolization of phage inside the cabinet yielded an average of 9 x 10 PFU per ft(3) outside; an average of 4.1 x 10(6) PFU per ft(3) were detected with the fans not in operation, a reduction of more than four logs. In-use studies on effectiveness showed that the cabinet significantly reduced the incidence of mice originally titer-free to Reo-3 virus. Hemagglutination inhibition antibodies to Reo-3 were detected in 9/22 (42%) mice housed in a conventionally ventilated animal laboratory while no seroconversion was detected in any of 22 mice housed in the mass air flow cabinet in the same laboratory.     

Nonadhesive populations in cultures of mesenchymal stromal cells from hematopoietic organs in mouse and rat

The study of adhesive properties of multipotent mesenchymal stromal cells evaluated from fibroblast colony-forming units in the bone marrow of adult mice and rats in populations of cells attached and unattached to plastic substrate after 2 h to 7 days in culture demonstrated both similarities and differences. The increase in the fibroblast colony-forming units in the adhesive population peaked on day 7 of in vitro culture in both cases; however, nearly no fibroblast colony-forming units were observed in the nonadhesive population from the mouse bone marrow in this period. Conversely, the number of colonies from the rat bone marrow nonadhesive population on day 7 of culture considerably increased, and this nonadhesive population in long-term culture became the source for subsequent nonadhesive subpopulations containing fibroblast colony-forming units. After 7 days of in vitro culture, the suspension of cells isolated from the liver of 17-day-old rat fetuses also contained a fraction of unattached fibroblast colony-forming units. In the nonadhesive subpopulations from the bone marrow and fetal liver, fibroblast colony-forming units were observed up to day 48 and 30, respectively. Stromal cell precursors of nonadhesive subpopulations from the rat bone marrow featured a period of colony formation reduced to 7 days (i.e., they were formed 1.5-2 times faster compared to the primary culture). The total number of fibroblast colony-forming units from all nonadhesive subpopulations was roughly 6 and 7.4 times that of the adhesive population of the primary culture from the bone marrow and fetal liver, respectively. Considering that the mammalian bone marrow remains the preferred source of mesenchymal stromal cells, using nonadhesive subpopulations in the presented culture system can considerably increase the yield of stromal precursor cells     

Planktonic Marine Luminous Bacteria: Species Distribution in the Water Column

Luminous bacteria were isolated from oceanic water samples taken throughout the upper 1,000 m and ranged in density from 0.4 to 30 colony-forming units per 100 ml. Generally, two peaks in abundance were detected: one in the upper 100 m of the water column, which consisted primarily of Beneckea spp.; and a second between 250 and 1,000 m, which consisted almost entirely of Photobacterium phosphoreum. The population of P. phosphoreum remained relatively stable in abundance at one station that was visited three times over a period of 6 months. However, the abundance of luminous Beneckea spp. isolated from the upper waters fluctuated considerably; they were, as high as 30 colony-forming units per 100 ml in the spring and were not detected in the winter. Water samples from depths of 4,000 to 7,000 m contained less than 0.1 luminous colony-forming unit per 100 ml. The apparent vertical stratification of two taxa of oceanic luminous bacteria may reflect not only differences in physiology, but also depth-related, species-specific symbiotic associations.     

Survival of bacillus licheniformis on human skin.

The colonization and survival of Bacillus species, members of the cutaneous microbial community of humans, were investigated by applying spores of Bacillus licheniformis to the forearms of volunteers. Four strains were tested, including the bacitracin producer ATCC 10716 and its bacitracin-negative mutant. Germination occurred within 24 h. Significant differences in survival population and duration were found among the test strains; however, ATCC 10716 and its mutant produced statistically similar survival curves. In general, an inoculum density of 10(4) colony-forming units per cm2 allowed survival for at least 2 weeks. Individual variation was extreme, for one subject harbored bacilli for over 2 months and another eliminated the microorganism within 3 days. Individuals could be differentiated into long-term (greater than 21 days) and short-term (less than 14 days) carriers. Eight of the 11 volunteers (73%) inoculated with ATCC 10716 carried it for 2 weeks, and 5 subjects (45%) continued to support the bacilli for 3 weeks. Spreading of the organism to other regions of the body occurred, but bacilli were not detected in these areas beyond 6 days.     

Effect of radiation, thermal or combined radiation-thermal damage to a population of progenitor cells of the granulocyte-macrophage series in mouse bone marrow

The recovery of a population of granulopoietic-monocytic colony-forming units of mouse bone marrow was only slightly inhibited during the first two weeks after exposure to a mixture of radiation (4 Gy) and heat (burn of 10 per cent of the skin). In the interval between days 14 and 28 following the effect, their number reached the control level, then it decreased again.     

Staphylococci in the microbiocenosis of the skin of the hands]

The skin autoflora on the pad of a forefinger and the back and palm of a hand was studied in 40 healthy males aged 18-60 years by the modified washing and scraping method of P. Williamson and A. Kligman. 638 cultures of aerobic microorganisms, including coccal (55.3%) and bacilliform (44.7%) microbes, were isolated. In 6 persons (15%) coagulase-positive staphylococci were detected. Out of 10 coagulase negative species of this genus, S. epidermidis, S. saprophyticus and S. warneri occurred most frequently on the skin of hands. The highest density of bacterial populations (10.970 +/- +/- 1.845 cells/sq. cm) was registered on the back of hands, the surface of palms was found to have somewhat lower density (8.679 +/- 1.282 cells/sq. sm) and the skin of forefingers, the lowest density of bacterial populations (6.878 +/- +/- 1.137 cells/sq. sm). 17.5% of examined persons were found to be carriers with S. aureus isolated from their nasal mucosa. S. aureus isolated from the skin surface and the nasal cavity of different persons belonged to different phage variants, but S. aureus isolated from the nasal cavity and the skin of the same person belonged to one phage variant.     

Growth and differentiation potential of main- and side-population cells derived from murine skeletal muscle

Skeletal muscle-derived CD34+/45- (Sk-34) cells were identified as a new candidate for stem cells. However, the relationship between Sk-34 cells and side-population (SP) cells is unknown. Here, we demonstrate that Sk-34 cells prepared from murine skeletal muscles consist wholly of main-population (MP) cells. The Sk-34 cells included only a few SP cells (1:1000, SP:MP). Colony-forming units of Sk-34 cells of both SP and MP possessed the same potential to differentiate into adipocytes, endothelial, and myogenic cells and showed the same colony-forming activity (1.6%). In addition, the colony-forming units of the CD34-/45- (double negative: DN) population were found to begin CD34 expression and to possess the potential to differentiate into myogenic and endothelial cells. We also found that expression of CD34 antigen precedes MyoD expression during the myogenic process of DN cells. Furthermore, both Sk-34 and DN cell populations were mostly negative for CD73 (93-95%), whereas the CD45+ cell population was >25% positive for CD73, and this trend was also seen in bone marrow-derived CD45+ cells. These results indicate that the MP cell population is about 99.9% responsible for the reported in vitro myogenic-endothelial responses of skeletal muscle-derived cells.     

The vertical distribution of macrobenthos within the substratum of the Brazos river,Texas

Vertical stratification samplers were developed for sampling the grave-sand substrate of a Brazos River, Texas riffle. Fifteen of 25 species recovered, occurred below 10 cm. Mean percentages of total organisms recovered were 66.4%, 20%, 6.1% and 7.5% per 10 cm level, respectively, from the surface down. Dominant insects were Neochoroterpes mexicanas naiads and chironomid, Simulium, Cheumatopsyche and Stenelmis larvae. Seasonal population peaks of these five groups in the top 10 cm correspond with observed emergence peaks. The smaller size classes were generally predominant in the 0–10 cm level. Larvae of Stenelmis were the most evenly distributed among the various 10 cm levels in all size classes. A movement of Cheumatopsyche and Neochoroterpes to lower levels was observed following a large flood, suggesting an escape response to increased silt load and scouring.Dissolved oxygen ranged from saturation at the surface down to 0.4–0.7 ppm at 30–40 cm, indicating that it was possibly limiting at lower levels. Maximum temperature difference between to cm levels was only 3 C. Flow was negligible below to cm.The vertical stratification sampler recovered significantly greater populations in the surface 20 cm, but not in the total 40 cm, than a modified Hess sq. ft. sampler.Study supported in part by the Faculty Research Fund and Institute for Environmental Studies of North Texas State University.     

The numbers and distribution of buffalo in the Ruwenzori National Park*, Uganda

The buffalo (Syncerus coffer (Sparrman)) in the Ruwenzori National Park, Uganda were censused by means of aerial survey. Details of earlier censuses of buffalo from the air are given. Herds were located by flying along flight lines 1–6 km apart at a height of 300 m above ground. Buffalo in small herds and in bachelor groups containing less than thirty animals were counted individually but most herds were photographed and the number of buffalo counted from the prints. Two full photographic counts were made in November 1968 and December 1969 and subsequently, counts of herds only were made four times a year during each wet and dry season. The total numbers of buffalo were estimated from the herd counts on the basis of the mean herd size and percentage of bachelors recorded earlier. A total of 17 835 buffalo, comprising 16 749 in 165 herds and 1086 bachelor bulls, was recorded in 1968 and 18 040 buffalo made up of 17 141 in 162 herds and 899 bachelors were counted in 1969. The mean herd size was 101-5 in 1968 and 105-8 in 1969 with bachelors representing 6-1% and 5-0% of the totals in 1968 and 1969 respectively. The mean size of the bachelor groups was 4–7 and 3-3 in 1968 and 1969 respectively. The density of the buffalo was the same each year at twelve animals per sq. km. If only the preferred habitat is considered, the density becomes thirty-eight buffalo per sq. km. The mean areas occupied by a single herd were 9-4 and 9-6 sq. km in 1968 and 1969 respectively. The mean area of preferred habitat used by one herd was 2–9 sq. km each year. It is concluded that herd counts are not a satisfactory method for assessing the total buffalo population although they have value as indices, provided the same observer is used. However, they do suggest in the present work that there is a slight but definite seasonal movement of buffalo into forested areas or towards permanent water during the dry season.     

Estimation of the absolute density of adult Ixodes persulcatus ticks according to the results of collecting per flag-hour (Ixodidae)

The results of two methods of estimation of abundance of adult Ixodes persulcatus ticks, the absolute (at the sample plots) and the relative (per flag-hour) estimations, were compared. Collecting of ticks and estimation of their abundance was conducted during 9 years at the forests of the Far East and Pre-Ural region. The total of 1995 plots (100 sq. m each) were studied and 865 flag-hours were carried out. A good correlation of the data, obtained by these methods, was revealed. The average number of adult ticks, collected per flag-hour, approximately corresponds to the average number of ticks, activated during the season at 100 sq. m. The possibility of corresponding re-calculation of the results of estimation per flag-hour into the parameters of tick population density was evaluated. It was shown that such re-calculation gives good results. Their inexactitude usually doesn't exceed the statistical error of the parameters, obtained by the estimation of tick density at sample plots.     

Effect of Urea Concentration on Growth of Ureaplasma urealyticum (T-Strain Mycoplasma)

The effect of urea on growth of Ureaplasma urealyticum type VIII was studied by cultivating the organisms in a dialysate broth, prepared from soy peptone and autoclaved yeast, supplemented with 5% dialyzed horse serum, 100 mM 2-(N-morpholino)ethane sulfonic acid buffer (pH 5.75), and defined amounts of urea. Without urea, growth did not occur. Total growth was directly related to urea concentration. The least amount of urea that supported growth was 0.032 mM, which resulted in 3 × 10⁴ colony-forming units per ml. The maximum yield of organisms, 8.0 × 10⁷ colony-forming units per ml, was observed at 32 mM urea. Growth was limited not only by urea concentration, but also by the buffer capacity of the medium. The maximum amount of 2-(N-morpholino)ethane sulfonic acid buffer that could be employed was 100 mM; at higher concentrations, growth was inhibited. The yield of U. urealyticum was small even in medium with 32 mM urea and 100 mM 2-(N-morpholino)ethane sulfonic acid buffer: 0.63 mg of protein per liter of culture containing 5 × 10¹⁰ total colony-forming units. The molar growth yield was 20 mg of protein per mol of urea. The growth rate was also a function of urea concentration. Generation times ranged from 8 h at 0.032 mM urea to 1.6 h at 3.2 mM urea, where the substrate level was saturating. The K_s value for growth was 2.0 × 10⁻⁴ M urea. Thus, urea is a growth-limiting factor for U. urealyticum, but remarkably large amounts of this substrate are required.     

Staphylococcal enterotoxin production in the presence of non-enterotoxigenic staphylococci.

Enterotoxigenic Staphylococcus aureus strains were grown with a non-enterotoxigenic strain in laboratory medium, in milk, and in ham. Differences in pigmentation were used to differentiate the enterotoxigenic strains from the non-enterotoxigenic ones. Enterotoxin was detectable in milk when the colony counts of the non-enterotoxigenic strain were 15 to 20 times greater than those of the enterotoxigenic ones and in ham when the ratio was 60 to 77:1. Enterotoxin was detectable in milk when the enterotoxigenic strains reached counts of 10(7) colony-forming units per ml and in ham when the counts reached 10(8) colony-forming units per ml. It may be necessary in some food poisoning outbreaks to examine many isolates (up to 50 or 60) for enterotoxin production to be able to detect the enterotoxigenic staphylococci.     

Long-term Field Efficacy of the Entomogenous Fungus Metarhizium anisopliae against the Subterranean Scarab, Adoryphorus couloni

The efficacy of Metarhizium anisopliae DAT F-001 against Adoryphorus couloni was examined over 4 years on a sheep grazing property ('Inverell') in central Tasmania. Three generations of the primary A. couloni population and two generations of the overlapping population were studied. Application of 5.1 0.7 104 M. anisopliae spores g-1 of soil (2 cm below the soil surface), during midwinter 1989, when the primary population was in the middle of the L3 larval stage, resulted in 30.3% fewer larvae in treated plots by 21 weeks and 57.8% less pupae by 27 weeks. The population decline was consistent with a mortality model developed from laboratory data. In the two subsequent generations of the primary population, there were 63.2% (1991) and 45.0% (1993) fewer larvae in the treated plots before the damaging L3 stage. The two sequential generations of the overlapping A. couloni larval populations had 68% (1990) and 65% (1992) fewer larvae in the treated plots than in the untreated plots. Reductions in larval numbers led to a greater retention of sown perennial grasses, reduced weed invasion and a 23% increase in pasture productivity in the autumn of 1992. Incorporation of M. anisopliae into the soil did not reduce the numbers of non-target invertebrates. The level of M. anisopliae DAT F-001 in the pasture remained at levels close to the applied concentration (5.1 0.7 colony-forming units g-1 of soil), but increased 10-fold when mummified L3 larval, prepupal and pupal cadavers were present from January to June 1990. The level of the fungus in the soil was still twice the original applied concentration at the conclusion of sampling in March 1992.