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Monitoring the bacterial population dynamics during fermentation of artisanal Argentinean sausages.

The dynamics of the microbial community responsible for the artisanal fermentation of dry sausage produced in Argentina was investigated by using classical and molecular approaches. The combined use of RAPD analysis with primers M13, XD9, RAPD1 and RAPD2 and 16S rDNA sequencing were applied to the identification and intraspecific differentiation of 100 strains of lactobacilli and Micrococcaceae. DGGE analysis was used to monitor the dynamic changes in population after total microbial DNA was directly extracted from sausages and subjected to PCR using V3f (GC), Bact-0124f-GC and Univ-0515r primers. The sequence analysis of 16S rDNA of the dominant species was also carried out. Lactobacillus sakei and Lactobacillus plantarum were the dominant lactic acid organisms during the fermentation while Staphylococcus saprophyticus represented the dominant species of Micrococcaceae. It was demonstrated that the ripening process of Argentinean artisanal fermented sausage is driven by a limited number of Lactobacillus and Staphylococcus strains selected from environmental microbiota by the ability to best compete under the prevailing conditions of the ecological niche. The identification of dominant communities present in this artisanal fermented sausage can help in the selection of starter cultures consisting in well adapted strains to the particular production technology.

Animals↗

Numbers and types of microorganisms in vacuum-packed cold-smoked freshwater fish at the retail level.

Fifty-four packages (each one belonging to a different lot) of vacuum-packed cold-smoked salmon (30) and trout (24) produced by six Spanish smokehouses were obtained at retail level after 3 weeks storage at 2+/-1 degrees C. Sensorial, chemical, physicochemical and microbiological characteristics were examined. Overall, pH, a(w), salt content in water phase, aerobic plate counts at 30 and 25 degrees C. levels of Enterobacteriaceae, lactic acid bacteria (LAB), fungi and presumptive aeromonads and staphylococci are in agreement with available data on lightly preserved fish products. Psychrotrophic clostridia ranged between 1.71 and 2.21 log CFU/g. Levels of ethanol were highly variable and not significantly related (p > 0.05) to sensory scores or to microbial numbers. Salmonella, Escherichia coli and Listeria monocytogenes were not detected in any sample. Listeriae other than L. monocytogenes were isolated from three packages. Levels of Staphylococcus aureus lower than 4 log CFU/g were also found in three packages. Among 377 bacteria randomly isolated from aerobic 25 degrees C plate counts, LAB predominated, with Carnobacterium (C. piscicola) and Lactobacillus (eight species) being the genera most frequently found. The second and third major groups were Enterobacteriaceae and Micrococcaceae, respectively. Proteus vulgaris, P. mirabilis and Serratia liquefaciens were dominant among Enterobacteriaceae and coagulase-negative staphylococci among Micrococcaceae. Minor microbial groups such as aerobic gram-negative bacilli (Acinetobacter; Moraxella and Pseudomonas), Brochothrix, Aeromonas, Bacillus and Vibrio constituted less than 17% of the total flora.

Animals↗

Water and air in two poultry processing plants' chilling facilities--a bacteriological survey.

1. Water, aerosols and air in chilling facilities in 2 poultry abattoirs were bacteriologically examined. The different types of samples displayed a variety of bacterial loads and genera. 2. The aerobic plate count in water from spray nozzles was about log10 2 to 3 in both plants. Flavobacterium, Alcaligenes, Acinetobacter and Pseudomonadaceae were predominant in these samples. 3. The microbial load in air of the 2 plants was quite different, consisting mainly of Micrococcaceae and Gram-positive irregular rods. 4. The aerobic plate count in aerosols was extremely high. The flora was the same as was found in the spray nozzles samples, plus Enterobacteriaceae, Micrococcaceae and streptococci, which presumably came from the air in the system. Cross-contamination may well occur in evaporative chilling.

Abattoirs↗

Heat inactivation of staphylococcus epidermidis at various water activities.

Members of the family Micrococcaceae play an important role in food spoilage and even in food poisoning. In contrast to members of the family Enterobacteriaceae, these bacteria can grow in media with low water activities. Therefore, the heat resistance of Staphylococcus epidermidis, a rather resistant member of the family Micrococcaceae, was studied at water activities between 0.87 and 1.00. The heat inactivation curves were clearly biphasic at all temperatures and water activities tested. Especially at low water activities, the D-values of the tail phase were extremely high (at 0.87 water activity, a D-value at 70 degrees C of 500 s was recorded).

Hot Temperature↗

Numerical survey of some bacterial taxa.

Focht, D. D. (Iowa State University, Ames), and W. R. Lockhart. Numerical survey of some bacterial taxa. J. Bacteriol. 90:1314-1319. 1965.-A numerical analysis was made of 77 properties of each of 43 bacterial strains, representing 25 genera from 8 families in the orders Eubacteriales and Pseudomonadales. Four major groups were found, related to one another at approximately the same level of similarity: (1) a large cluster containing the subgroups (1a) Athiorhodaceae-Spirillaceae, (1b) Xanthomonas, and (1c) "inactive" Micrococcaceae-Achromobacteraceae; (2) a cluster containing the "active" Micrococcaceae and Lactobacillaceae; (3) the enterobacteria; and (4) Aeromonas. There was a sharp distinction between the branches of groups 1a, 1c, and 2. The composition of groups was essentially the same whether or not fermentation of carbohydrates (28 characters) was included in the analysis. Several individual strains, notably, Bacillus subtilis, Leuconostoc mesenteroides, Pseudomonas aeruginosa, and Erwinia amylovora, were related to none of the groups, and others (two species of Proteus, Flavobacterium devorans, and Lactobacillus casei) showed only minimal quantitative relationships with their groups. These results suggest that there may be significant variation in levels of similarity within microbial groups presently accorded equivalent taxonomic rank, and that some present distinctions among taxa, particularly at the generic level, cannot be confirmed on the basis of overall similarity.

Bacteria↗

Genetic relationship between Sarcina ureae and members of the genus Bacillus.

Deoxyribonucleic acid-ribonucleic acid (DNA-RNA) and DNA-DNA hybridization studies were performed to determine the degree of genetic relatedness between Sarcina ureae and several members of the genus Bacillus. DNA-RNA hybridization showed a high degree of homology between S. ureae RNA and DNA from Bacillus species having a similar guanine plus cytosine content. The DNA from other genera of the family Micrococcaceae showed less homology with S. ureae RNA than did that of the Bacillus species tested; however, this homology was not found between the DNA of S. ureae and DNA from these Bacillus species or DNA from the other Micrococcaceae tested. Transformation with Bacillus DNA, infection with representatives from several major classes of Bacillus phages, and electrophoretic analysis of proteins in crude extracts of these strains were also attempted as a further test of the genetic relationship between the genera. These experiments did not support the belief that the two groups are closely related genetically.

Bacillus↗

The classification of staphylococci from colonized ventriculo-atrial shunts.

Micrococcaceae isolated from the shunt, ventricles, and bloodstream of children with colonized ventriculo-venous shunts were classified within the scheme of Baird-Parker (1963). With one exception, all belonged to subgroup II of the genus Staphylococcus; tests were therefore devised for division within this subgroup, and results are reported in 30 cases from this and other hospitals. Skin and nasal staphylococci isolated from many of these patients were compared with those recovered from their shunts and blood. Evidence is offered for the occasional coexistence of more than one strain of staphylococcus in colonized shunts and in the bloodstream. Successive recolonization of replaced shunts was apparently not necessarily caused by the same type of staphylococcus. Nasal and skin micrococcaceae from many other babies, both in hospital and in parental care, from hospital staff and from adults selected at random from non-hospital sources, were similarly classified. The validity and significance of the findings are discussed.

Adult↗

Evaluation of the spiral plating method for the enumeration of microorganisms throughout the manufacturing and ripening of a raw goat's milk cheese.

A statistical comparison of the spiral plate count (SPLPC) and the standard plate count (SPC) methods for enumeration of microorganisms in raw goat's milk cheese throughout its manufacturing and ripening was carried out. Enumeration of mesophiles, lactic acid bacteria (presumptive lactococci, presumptive leuconostocs, and presumptive lactobacilli), Micrococcaceae, Enterobacteriaceae, and molds and yeasts was carried out for milk, curd, and 2-, 5-, 10-, 17-, and 27-day-old cheeses. Average counts for the SPLPC and SPC methods differed by less than half of a log cycle for all microbial groups studied (range of difference, -0.1386 [mesophiles] to +0.4397 [presumptive lactobacilli]). The results of the SPLPC method compared favorably with the results of the SPC procedure for mesophiles, presumptive lactococci, presumptive leuconostocs, Enterobacteriaceae, and molds and yeasts (the variance between replicate platings was close to 0.005, and correlation coefficients were >0.9). Correlation coefficients were lower for Micrococcaceae (r = 0.824) and presumptive lactobacilli (r = 0.670). Analysis of variance showed that the plating method was a significant factor (P < 0.05) for presumptive lactobacilli counts. In general, results from the SPLPC method compared favorably with results from SPC procedure in the enumeration of microorganisms in goat cheese throughout its manufacturingand ripening processes. However, the suitability of the SPLPC method depends mainly on the microbial group studied.

Animals↗

[Topical treatment of acne vulgaris with erythromycin and benzoylperoxide (author's transl)].

16 healthy persons were treated for 7 weeks with a 1% erythromycin containing ointment. Another 25 persons (among them 10 acne patients) were treated during the same period of time with a combination therapy consisting of erythromycin (1% in an unspecific ointment base) and benzoylperoxide (5% in a gel base). The combined therapy reduces the micrococcaceae in the pilosebaceous ducts significantly. On the other side the therapy consisting only of erythromycin causes only initially a decrease and then an increase in the number of micrococcaceae. Induction of resistance to erythromycin is in both cases considerable. In both cases a reduction of propionibacteria which are most important for the pathogenesis of acne must be assumed. Because of the probable pathogenetic role of staphylococci in acne vulgaris an additional improvement of the therapeutical results can be expected in the case of combination therapy. In vitro experiments demonstrate further that an erythromycin treatment leads to considerable resistance induction in S. aureus, regardless of whether it is applied alone or in combination with benzoyl peroxide.

Acne Vulgaris↗

Urinary tract infections due to Staphylococcus saprophyticus biotype 3.

Staphylococcus saprophyticus biotype 3 (Micrococcus subgroup 3 or M3) has usually been shown to be the second commonset cause of urinary tract infections in European women who are not in hospital. It generally causes pyuria and symptoms as severe as those caused by Escherichia coli. Unlike S. epidermidis it is seldom found as a contaminant in midstream urine specimens, and almost exclusively infects women in their reproductive years. However, S. saprophyticus is seldom differentiated from S. epidermidis in Canadian clinical laboratories. Urinary isolates of S. saprophyticus were presumptively differentiated from other coagulase-negative Micrococcaceae by their resistance to novobiocin as demonstrated by a simple disc susceptibility test that misidentified the infecting organism in only 3.4% of specimens. These novobiocin-resistant, coagulase-negative organisms caused similar proportions of the urinary tract infections in young women in York, England and Vancouver -- 6.6% and 6.9% respectively. In York these organisms were associated with significantly greater pyuria than novobiocin-sensitive organisms or bile-tolerant streptococci but not S. aureus or Enterobacteriaceae. In both communities novobiocin-sensitive, coagulase-negative Micrococcaceae were appreciably more resistant to penicillin than novobiocin-resistant organisms. Thus, differentiating S. saprophyticus from novobiocin-sensitive, coagulase-negative organisms provides information that is clinically useful, particularly for primary care practitioners working in the community or in outpatient clinics.

Adult↗

Effect of natural infection with minor pathogens on susceptibility to natural infection with major pathogens in the bovine mammary gland.

OBJECTIVE: To evaluate the effect of natural udder infection with minor pathogens on subsequent natural infection with major pathogens. SAMPLE POPULATION: 7 dairy herds with low bulk milk somatic cell count. PROCEDURE: During a 20-month prospective study, milk samples were collected from diary cows at regular intervals and from quarters with clinical signs of mastitis. Incidence of intramammary infection was calculated in uninfected quarters and in quarters infected with minor pathogens. A within-cow, matched case-control analysis was used to evaluate the effect of minor pathogens on subsequent infection with major pathogens. RESULTS: Quarters infected with minor pathogens had higher somatic cell count than did uninfected quarters. In quarters infected with Corynebacterium bovis, the rate of infection with major pathogens was lower, whereas in quarters infected with coagulase-negative Micrococcaceae, the rate of infection with major pathogens was higher than that in uninfected quarters. From the within-cow comparison, it appeared that, in quarters infected with minor pathogens, infection with major pathogens was significantly lower than that in comparable control quarters not infected with minor pathogens. CONCLUSIONS: Minor pathogens have a protective effect against infection with major pathogens. The protective effect of C bovis against subsequent infection with major pathogens appears to be greater than the effect of coagulase-negative Micrococcaceae.

Animals↗

Investigation of the mechanism of action of 2% fusidic acid lotion in the treatment of acne vulgaris.

We describe the results of a single-centre, double-blind, vehicle-controlled, parallel group study on the quantitative effects of 2% fusidic acid lotion (Fucidin lotion) in facial acne vulgaris. The trial was completed by 52 patients aged 15-25 years with mild to moderate acne who had been randomized to either Fucidin Lotion (n = 25) or its base (n = 27). Primary outcome measures included colony counts of Propionibacterium acnes and micrococcaceae and measurements of skin surface lipid free fatty acids and sebum excretion rate. Clinical assessment was based on the acne grade, count of inflamed and non-inflamed lesions and evidence of a primary irritant dermatitis. There was a variable but gradual reduction in lesion counts with the maximum improvement at 12 weeks for inflamed lesions, where the reduction was 19.9% for fusidic acid and 24.7% for the placebo. The non-inflamed lesions decreased by 10.8% in the fusidic acid group and increased by 15.9% in the placebo group; this difference was not statistically significant. Although the fusidic acid reduced the micrococcaceae count by 1 log cycle, inferring adequate compliance, there was no reduction in the counts of P. acnes, surface free fatty acids or sebum excretion rate. This study has failed to explain the mechanism of action of topical fusidic acid.

Acne Vulgaris↗

Qualitative and quantitative investigations on the resident bacterial skin flora in healthy persons and in the non-affected skin of patients with seborrheic Eczema.

The composition of the aerobic and anaerobic bacterial skin flora was investigated quantitatively and qualitatively in 25 patients with seborrheic eczema (SE) and in 35 healthy persons. In SE patients, the geometric mean count for propionibacteria in the pilosebaceous ducts of the forehead was reduced to a statistically significant extent in comparison with the control group. There was the same tendency with regard to the geometric mean count of the superficial anaerobic bacterial flora of the back, whereas the geometric mean counts of the coagulase-negative staphylococci were the same in the two groups. These results may explain earlier findings of a reduction of free fatty acids in the surface lipids in SE patients. According to the results of differentiation in 295 propionibacteria and 157 micrococcaceae, 7 biotypes and 10 phage types of propionibacteria which were so far unknown could be demonstrated. Significant differences between the groups with regard to the frequency of different biotypes, serotypes, and phage types could be demonstrated neither in propionibacteria nor in coagulase-negative staphylococci. The results of typing, however, showed that patients with SE displayed much more frequently several different propionibacterial species and types as well as different micrococcal species and types than persons with healthy skin.

Adult↗

Unstable L-forms of micrococci in human foetal blood.

In human foetal blood the presence of Micrococcaceae in the unstable L-form, probably taking origin from the placental transmission of minimal reproductive units, has been recognized by means of microscopic and cultural methods.

Fetal Blood↗

Investigation of the hygienic safety of continuous positive airways pressure devices after reprocessing.

With the widespread use of continuous positive airways pressure (CPAP) therapy, the safety of CPAP devices after reprocessing is the subject of debate. In this study, the contamination of CPAP devices and the effectiveness of disinfection was investigated. A total of 122 CPAP devices were examined including 50 CPAP devices used by patients, which were examined before and after reprocessing. Seventy-two new CPAP devices that had not been in contact with patients served as controls. The results of this study show that the microbial contamination of new and used CPAP devices is only minimal. Contaminating micro-organisms were predominantly micro-organisms reflecting the normal environmental microflora such as Penicillium spp., Aspergillus spp., Micrococcaceae and Bacillaceae. Gram-negative species could only be found in rare cases. The data obtained give no indication of poor disinfection of CPAP devices.

Air Microbiology↗

Stomatococcus mucilaginosus as an agent of CAPD peritonitis.

We report a case of bacterial CAPD peritonitis in a 57 year old immunosuppressed woman with renal insufficiency caused by a rare species of the Micrococcaceae, Stomatococcus mucilaginosus. This uncommon case shows that the presence of gram-positive cocci in pairs, tetrads and clusters forming whitish-gummy colonies should remind us of the possibility of Stomatococcus mucilaginosus.

Bacterial Infections↗

[Stomatococcus mucilaginosus: cultural and biochemical properties of 100 strains isolated from the ORL area].

A study of the growth characteristics and 80 biochemical tests using API System micromethods (API-50CH, API-20E, API-Zym) was carried out on 100 strains of Stomatococcus mucilaginosus isolated from the human oral cavity. Seventy-seven other enzymatic tests from the API system (AP-arylamidases, osidases and esterases) were also performed on 10 of these strains. The results of these tests were compared to those of 24 type strains of Micrococcaceae and Aerococcus viridans. A simple identification scheme for S. mucilaginosus is proposed.

Adult↗

In-vitro and in-vivo efficacy of zinc acetate against propionibacteria alone and in combination with erythromycin.

Some studies have been published about the in vitro activity of zinc acetate (ZA), erythromycin (E) and their combination (ZA/E) against Propionibacterium spp., especially erythromycin resistant strains. The efficacy of topical ZA/E combination has been reported as well, but a comparison to ZA monotherapy is missing. Therefore, the MIC values of ZA, E and the ZA/E combination were determined for 15 erythromycin-resistant and 12 erythromycin-sensitive Propionibacterium strains using the agar dilution method and the checkerboard technique. Furthermore, the antimicrobial efficacy of ZA (1.2%) vs. the ZA/E (1.2%/4%) combination in an alcoholic solution was tested in a 7-day treatment administered to 32 acne patients. The MIC 100 for ZA was 1024 micrograms ZA/ml for both, erythromycin resistant and erythromycin sensitive Propionibacterium strains. The ZA, as well as the ZA/E solution showed efficacy reducing both the Propionibacterium spp., and the Micrococcaceae in the sebaceous gland infundibula of acne patients. There was no significant difference between the two treatments. As the MIC 100 of ZA/E was equal to the MIC 100 of ZA, the decrease of the erythromycin MIC of the ZA/E combination in erythromycin-resistant strains may be partly attributed to the addition of ZA to E. The in vivo antibacterial efficacy on 32 acne patients supports the hypothesis that the antibacterial effect of ZA/E in short-term treatment can be mostly attributed to ZA.

Acne Vulgaris↗