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Fate of Cryptosporidium oocysts, Giardia cysts, and microbial indicators during wastewater treatment and anaerobic sludge digestion.

The extent of reduction in selected microorganisms was tested during both aerobic wastewater treatment and anaerobic digestion of sludge at the wastewater treatment plant in Ottawa to compare the removal of two encysted pathogenic protozoa with that of microbial indicators. Samples collected included the raw wastewater, the primary effluent, the treated wastewater, the mixed sludge, the decanted liquor, and the cake. All of the raw sewage samples were positive for Cryptosporidium oocysts and Giardia cysts, as well as for the other microorganisms tested. During aerobic wastewater treatment (excluding the anaerobic sludge digestion), Cryptosporidium and Giardia were reduced by 2.96 log10 and 1.40 log10, respectively. Clostridium perfringens spores, Clostridium perfringens total counts, somatic coliphages, and heterotrophic bacteria were reduced by approximately 0.89 log10, 0.96 log10, 1.58 log10, and 2.02 log10, respectively. All of the other microorganisms were reduced by at least 3.53 log10. Sludge samples from the plant were found to contain variable densities of microorganisms. Variability in microbial concentrations was sometimes great between samples, stressing the importance of collecting a large number of samples over a long period of time. In all cases, the bacterial concentrations in the cake (dewatered biosolids) samples were high even if reductions in numbers were observed with some bacteria. During anaerobic sludge digestion, no statistically significant reduction was observed for Clostridium perfringens, Enterococcus sp., Cryptosporidium oocysts, and Giardia cysts. A 1-2 log10 reduction was observed with fecal coliforms and heterotrophic bacteria. However, the method utilized to detect the protozoan parasites does not differentiate between viable and nonviable organisms. On the other hand, total coliforms and somatic coliphages were reduced by 0.35 log10 and 0.09 log10, respectively. These results demonstrate the relative persistence of the protozoa in sewage sludge during wastewater treatment.

Animals↗

Comparison of different regimens for surgical hand preparation.

Twenty surgical staff members participated in a clinical trial to compare the microbiology and skin condition of hands when using a traditional surgical scrub (TSS) with a detergent-based antiseptic containing 4% chlorhexidine gluconate (CHG) and a short application without scrub of a waterless hand preparation (HP) containing 61% ethyl alcohol, 1% CHG, and emollients. The HP was associated with less skin damage (P = .002) and lower microbial counts postscrub at days five (P = .002) and 19 (P = .02). The HP protocol had shorter contact time (HP mean [M] = 80.7 seconds; TSS M = 144.9 seconds; P < .0001), and more subjects preferred the HP regimen (P = .001). The HP performed better than the TSS, was less costly, and should be evaluated in larger trials and considered for widespread implementation.

Adult↗

Skin flora of patients in Thailand.

BACKGROUND: Studies taken from different hospitalized patient populations, environments, and geographic regions reveal differences in the numbers and species of organisms colonizing the skin. Our aim was to determine the types and amounts of skin flora, and examine the factors associated with variations in microbial skin flora in patients in Thailand. METHOD: We studied 350 outpatients and 500 inpatients at Siriraj Hospital in Bangkok, Thailand. The skin at the forearm and the sternum of each patient was cultured by contact plate technique. RESULTS: The number of skin flora colony-forming units (CFUs) were correlated to the site of sampling. There was a significant correlation of CFUs between samples from the forearm and the sternum in patients who were hospitalized (r = 0.6; P <.001) and in outpatients (r = 0.5; P <.001). The numbers of micro-organisms on the sternum was significantly greater than the number cultured from the forearm for all patients. Inpatients had significantly more organisms on the the forearm and sternum compared with outpatients. High counts (CFUs > 600) were found more frequently in patients who were hospitalized; had chronic obstructive pulmonary disease, diabetes mellitus, or autoimmune diseases; and were undergoing operation and receiving antibiotics. Acinetobacter spp and methicillin-resistant Staphyloccoccus aureus were found more frequently in patients who were hospitalized. CONCLUSION: Skin flora of patients in tertiary care hospitals in Thailand has higher CFUs, and A baumannii is prevalent, especially in patients who are hospitalized.

Adult↗

Microbial contamination of hydrophilic contact lenses. Part I: Quantitation of microbes on patient worn-and-handled lenses.

We cultured Soflens (polymacon) contact lenses to determine the number of microorganisms present following normal patient wear and handling just prior to disinfection. Total protein deposited was determined for the companion lens from each patient. A random population of 109 adapted soft contact lens patients participated in the study. Some patients participated more than once, resulting in a total of 196 lenses being cultured and 195 lenses analyzed for total protein. The left lens was cultured immediately. The right lens was extracted at 70 degrees C in sodium hydroxide, and the total protein in the extract determined using a modified Lowry protein assay. The mean protein deposition per lens was 3.4 micrograms (median 2 micrograms per lens; range less than 1 to 78 micrograms/lens). Microorganisms were cultured from 95% of the lenses. The mean bacterial count (in colony forming units per lens) was 2,482 (median: 123; range less than 3 to 150,000). Fungal contamination was found on 11% of the lenses at very low levels (3-9 yeast/lens and 3-18 mold/lens). Statistical analysis found no significant relationship between bacterial bioburden and any of the study parameters, including total protein, lens age, or subjective evaluation of lens cleanliness.

Bacteria↗

Microbial quality of shrimp products of export trade produced from aquacultured shrimp.

Bacteriological quality of individually quick frozen (IQF) shrimp products produced from aquacultured tiger shrimp (Penaeus monodon) has been analysed in terms of aerobic plate count (APC), coliforms, Escherichia coli, coagulase-positive staphylococci, Salmonella, and Listeria monocytogenes. Eight hundred forty-six samples of raw, peeled, and deveined tail-on (RPTO), 928 samples of cooked, peeled, and deveined tail-on (CPTO), 295 samples of headless, undeveined shell-on (HLSO), and 141 samples of raw, peeled, and deveined tail-off (RPND) shrimps were analysed for the above bacteriological parameters. Salmonella was isolated in only one sample of raw, peeled tail-on. Serotyping of the strain revealed that it was S. typhimurium. While none of the cooked, peeled tail-on shrimp samples exceeded the aerobic plate count (APC) of 10(5) colony forming units per gram (cfu/g), 2.5% of raw, peeled, tail-on, 6.4% of raw, peeled tail-off, and 7.5% of headless shell-on shrimp samples exceeded that level. Coliforms were detected in all the products, though at a low level. Prevalence of coliforms was higher in headless shell-on (26%) shrimps followed by raw, peeled, and deveined tail-off (19%), raw, peeled tail-on (10%), and cooked, peeled tail-on (3.8%) shrimps. While none of the cooked, peeled tail-on shrimp samples were positive for coagulase-positive staphylococci and E. coli, 0.6-1.3% of the raw, peeled tail-on were positive for staphylococci and E. coli, respectively. Prevalence of staphylococci was highest in raw, peeled tail-off (5%) shrimps and the highest prevalence of E. coli (4.8%) was noticed in headless shell-on shrimps. L. monocytogenes was not detected in any of the cooked, peeled tail-on shrimps. Overall results revealed that the plant under investigation had exerted good process control in order to maintain superior bacteriological quality of their products.

Aquaculture↗

On-line sterilization of cheese whey using ultraviolet radiation.

The effectiveness of ultraviolet radiation for on-line sterilization of cheese whey was investigated. The effects of flow rate and residence time on the performance of three UV reactors having different gap sizes (18, 13, and 6 mm) were studied. Six flow rates and six residence times were tested with the three UV reactors. The cheese whey used in this study had a very high turbidity (4317 NTU), very poor transmittance in the UV radiation germicidal range ( approximately 0%), and high percentage of large solid particles ( approximately 20% > 100 microm). Although the cheese whey physical characteristics showed low probability of sterilization using UV radiation, the study showed that UV radiation can be used on-line to sterilize cheese whey if the proper reactor gap size and the appropriate residence time are used. There were combined effects of the flow rate and gap size. The cell removal efficiency increased with increases in residence time and decreases in the UV reactor gap size. Removal efficiency of 100% was not achieved in this study with the first UV reactor (18-mm gap size), whereas 100% removal efficiency was achieved with the second (13-mm gap size) and third (6-mm gap size) UV reactors at residence times of 2.0 and 0.5 h, respectively. The microbial decay rates achieved in this study were 4.94, 7.62, and 20.9 h(-)(1) using the first, second, and third UV reactor, respectively. Residence times of 3.3, 2.1, and 0.8 h would be required to completely destruct a microbial population of 5.95 x 10(6) cells/mL using the first, second, and third UV reactors, respectively. Although cheese whey sterilization using UV radiation seems to be a good alternative to pasteurization, increases in cheese whey temperature resulted in lamp fouling. If online sterilization is to be used, the fouling problem should be investigated and a maintenance scheme for the UV reactor should be developed.

Animals↗

Do intensive care drug infusions support microbial growth?

The potential for commonly infused drug solutions to support or inhibit microbial growth was explored in this study. Drugs examined were midazolam HCl, morphine sulphate, fentanyl citrate, pethidine HCl, bupivacaine HCl, atracurium besylate, vecuronium bromide, adrenaline, dopamine, dobutamine, noradrenaline, isoprenaline, glyceryl trinitrate, sodium nitroprusside and propofol. All drugs were studied in both diluted and undiluted forms. Of the 15 medications investigated, all except propofol were found to be bactericidal and to reduce fungal growth.

Anesthetics, Intravenous↗

[Causation of stomatitis during cytostatic chemotherapy for head and neck tumors].

In a prospective trial involving 17 patients with head and neck tumors an attempt was made to relate the occurrence of stomatitis during cytostatic chemotherapy to changes in the resident microbial population, if any. Of the 17 patients who received the same cytostatic regimen, 10 developed stomatitis after a mean interval of 12 days. Smears were taken of the 2 patient subgroups (with and without stomatitis) at the onset of treatment and after comparable intervals during treatment and compared to those of a normal control group. Pretreatment smears did not show any quantitative or qualitative differences in the aerobic and anaerobic microbial spectrum between the 3 subgroups examined. During treatment patients developing mucositis were found to show a statistically significant multiplication of pathogens, while the other 2 groups still did not differ from one another. Together with the direct cytostatic effect of chemotherapy, proliferation of pathogens in the oral cavity appears to contribute to the development of stomatitis.

Adult↗

Characterization of dominant microbiota of a Ghanaian fermented milk product, nyarmie, by culture- and nonculture-based methods.

AIMS: To characterize the predominant micro-organisms in a Ghanaian traditional fermented dairy product, nyarmie, made from cows' milk, using both culture- and nonculture-based methods. METHODS AND RESULTS: Samples of nyarmie were analysed from three production sites in Accra, by determining the counts on selective culture media. The microbial diversity occurring in nyarmie was also evaluated by 16S/18S ribosomal DNA PCR amplification and denaturing gradient gel electrophoresis. Results showed that nyarmie contained lactococci and lactobacilli in the range of 10(8) and 10(10) CFU ml(-1), respectively, and yeasts at around 10(7) CFU ml(-1). The pH ranged between 3.49 and 4.25. The predominant lactic acid bacteria (LAB) in nyarmie were Leuconostocmesenteroides ssp. mesenteroides, Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus, Lact.helveticus, Lact. delbrueckii ssp. lactis and Lactococcus lactis, while Saccharomyces cerevisiae was the predominant yeast species. Lactobacillus delbrueckii ssp. delbrueckii was not detected by cultivation but its predominance was revealed by PCR-DGGE analysis. CONCLUSIONS: The flora in products from different producers varied in the LAB composition present and may result in variations in product quality. SIGNIFICANCE AND IMPACT OF THE STUDY: Development and use of starter cultures for nyarmie may be beneficial in improving the consistency of product quality.

Animals↗

Aerobic spore-forming bacteria and chemical composition of some Nigerian fermented soup condiments.

A total of 97 strains of spore-forming Bacillus were isolated from 45 samples of three Nigerian fermented condiments, obtained from retail markets located in Southwestern Nigeria. The isolates were identified as B. subtilis (33%), B. pumilus (19%), B. licheniformis (22%), B. brevis (9%), B. megaterium (12%) and B. polymyxa (5%). The microbial load of the condiments showed that the average count of spore-formers was between 107 to 109 cfu/g. The moisture contents of iru, ugba and ogiri were 57.18%, 46.32% and 42.34%, respectively, while the protein contents were 18.26%, 17.17% and 17.96%. The percentage fat was 29.88%, 40.25% and 44.14% for iru, ugba and ogiri. The ash content ranged from 5.8 to 6.1%; a 0.1% titratable acidity and pH values above 7.0 were obtained for the three condiments.

Bacillus↗

Mold and endotoxin levels in the aftermath of Hurricane Katrina: a pilot project of homes in New Orleans undergoing renovation.

BACKGROUND: After Hurricane Katrina, many New Orleans homes remained flooded for weeks, promoting heavy microbial growth. OBJECTIVES: A small demonstration project was conducted November 2005-January 2006 aiming to recommend safe remediation techniques and safe levels of worker protection, and to characterize airborne mold and endotoxin throughout cleanup. METHODS: Three houses with floodwater lines between 0.3 and 2 m underwent intervention, including disposal of damaged furnishings and drywall, cleaning surfaces, drying remaining structure, and treatment with a biostatic agent. We measured indoor and outdoor bioaerosols before, during, and after intervention. Samples were analyzed for fungi [culture, spore analysis, polymerase chain reaction (PCR)] and endotoxin. In one house, realtime particle counts were also assessed, and respirator-efficiency testing was performed to establish workplace protection factors (WPF). RESULTS: At baseline, culturable mold ranged from 22,000 to 515,000 colony-forming units/m3, spore counts ranged from 82,000 to 630,000 spores/m3, and endotoxin ranged from 17 to 139 endotoxin units/m3. Culture, spore analysis, and PCR indicated that Penicillium, Aspergillus, and Paecilomyces predominated. After intervention, levels of mold and endotoxin were generally lower (sometimes, orders of magnitude). The average WPF against fungal spores for elastomeric respirators was higher than for the N95 respirators. CONCLUSIONS: During baseline and intervention, mold and endotoxin levels were similar to those found in agricultural environments. We strongly recommend that those entering, cleaning, and repairing flood-damaged homes wear respirators at least as protective as elastomeric respirators. Recommendations based on this demonstration will benefit those involved in the current cleanup activities and will inform efforts to respond to future disasters.

Air Microbiology↗

Prokaryotic metabolic activity and community structure in Antarctic continental shelf sediments.

The prokaryote community activity and structural characteristics within marine sediment sampled across a continental shelf area located off eastern Antarctica (66 degrees S, 143 degrees E; depth range, 709 to 964 m) were studied. Correlations were found between microbial biomass and aminopeptidase and chitinase rates, which were used as proxies for microbial activity. Biomass and activity were maximal within the 0- to 3-cm depth range and declined rapidly with sediment depths below 5 cm. Most-probable-number counting using a dilute carbohydrate-containing medium recovered 1.7 to 3.8% of the sediment total bacterial count, with mostly facultatively anaerobic psychrophiles cultured. The median optimal growth temperature for the sediment isolates was 15 degrees C. Many of the isolates identified belonged to genera characteristic of deep-sea habitats, although most appear to be novel species. Phospholipid fatty acid (PLFA) and isoprenoid glycerol dialkyl glycerol tetraether analyses indicated that the samples contained lipid components typical of marine sediments, with profiles varying little between samples at the same depth; however, significant differences in PLFA profiles were found between depths of 0 to 1 cm and 13 to 15 cm, reflecting the presence of a different microbial community. Denaturing gradient gel electrophoresis (DGGE) analysis of amplified bacterial 16S rRNA genes revealed that between samples and across sediment core depths of 1 to 4 cm, the community structure appeared homogenous; however, principal-component analysis of DGGE patterns revealed that at greater sediment depths, successional shifts in community structure were evident. Sequencing of DGGE bands and rRNA probe hybridization analysis revealed that the major community members belonged to delta proteobacteria, putative sulfide oxidizers of the gamma proteobacteria, Flavobacteria, Planctomycetales, and Archaea. rRNA hybridization analyses also indicated that these groups were present at similar levels in the top layer across the shelf region.

Antarctic Regions↗

A comparative heat inactivation study of indigenous microflora in beef with that of Listeria monocytogenes, Salmonella serotypes and Escherichia coli O157:H7.

AIMS: Thermal inactivation of a mixture of five strains of Listeria monocytogenes, four strains of Escherichia coli O157:H7 and eight serotypes of Salmonella were compared with that of indigenous microflora in 75% lean ground beef. METHODS AND RESULTS: Inoculated meat was packaged in bags that were completely immersed in a circulating water bath and held at 55, 57.5 and 60 degrees C for predetermined lengths of time. The surviving cell population was enumerated by spiral plating heat-treated samples onto tryptic soya agar supplemented with 0.6% yeast extract and 1% sodium pyruvate. D-values, determined by linear regression, in beef were 77.49, 21.9, and 10.66 min at 55, 57.5, and 60 degrees C, respectively, for indigenous microflora (z = 5.81 degrees C). When either of the three pathogens were heated in beef, their D-values calculated were significantly lower (P < 0.05) than those of indigenous microflora at all temperatures. The slope of the thermal death time curve for L. monocytogenes, E. coli O157:H7 and indigenous microflora were similar. Using a survival model for nonlinear survival curves, the D1-values at all temperatures for L. monocytogenes were significantly higher (P < 0.05) compared with those for Salmonella serotypes, E. coli O157:H7 or indigenous microflora. However, higher recovery of a subpopulation of the indigenous microflora in beef exposed to heating at 55, 57.5 or 60 degrees C resulted in significantly higher (P < 0.05) D2-values at all three temperatures, compared with those of the three pathogens at the same test temperatures. CONCLUSIONS: If the thermal process is designed to ensure destruction of indigenous microbial flora, it should also provide an adequate degree of protection against L. monocytogenes, Salmonella serotypes or E. coli O157:H7. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study will assist the retail food industry in designing acceptance limits on critical control points that ensure safety, without introducing pathogens in a retail food environment, against L. monocytogenes, E. coli O157:H7 and Salmonella in cooked ground beef.

Animals↗

In situ activity of a bacteriocin-producing Lactococcus lactis strain. Influence on the interactions between lactic acid bacteria during sourdough fermentation.

AIMS: To biochemically characterize the bacteriocin produced by Lactococcus lactis ssp. lactis M30 and demonstrate its effect on lactic acid bacteria (LAB) during sourdough propagation. METHODS AND RESULTS: A two-peptide bacteriocin produced by L. lactis ssp. lactis M30 was purified by ion exchange, hydrophobic interaction and reversed phase chromatography. Mass spectrometry of the two peptides and sequence analysis of the ltnA2 gene showed that the bacteriocin was almost identical to lacticin 3147. During a 20-day period of sourdough propagation the stability of L. lactis M30 was demonstrated, with concomitant inhibition of the indicator strain Lactobacillus plantarum 20, as well as the non-interference with the growth of the starter strain Lact. sanfranciscensis CB1. CONCLUSIONS: In situ active bacteriocins influence the microbial consortium of sourdough LAB and can "support" the dominance of insensitive strains during sourdough fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: The in situ bacteriocinogenic activity of selected lactococci enables the persistence of insensitive Lact. sanfranciscensis strains, useful to confer good characteristics to the dough, at a higher cell concentration with respect to other LAB of the same ecosystem.

Anti-Bacterial Agents↗

Effect of genetically modified Pseudomonas putida WCS358r on the fungal rhizosphere microflora of field-grown wheat.

We released genetically modified Pseudomonas putida WCS358r into the rhizospheres of wheat plants. The two genetically modified derivatives, genetically modified microorganism (GMM) 2 and GMM 8, carried the phz biosynthetic gene locus of strain P. fluorescens 2-79 and constitutively produced the antifungal compound phenazine-1-carboxylic acid (PCA). In the springs of 1997 and 1998 we sowed wheat seeds treated with either GMM 2, GMM 8, or WCS358r (approximately 10(7) CFU per seed), and measured the numbers, composition, and activities of the rhizosphere microbial populations. During both growing seasons, all three bacterial strains decreased from 10(7) CFU per g of rhizosphere sample to below the limit of detection (10(2) CFU per g) 1 month after harvest of the wheat plants. The phz genes were stably maintained, and PCA was detected in rhizosphere extracts of GMM-treated plants. In 1997, but not in 1998, fungal numbers in the rhizosphere, quantified on 2% malt extract agar (total filamentous fungi) and on Komada's medium (mainly Fusarium spp.), were transiently suppressed in GMM 8-treated plants. We also analyzed the effects of the GMMs on the rhizosphere fungi by using amplified ribosomal DNA restriction analysis. Introduction of any of the three bacterial strains transiently changed the composition of the rhizosphere fungal microflora. However, in both 1997 and 1998, GMM-induced effects were distinct from those of WCS358r and lasted for 40 days in 1997 and for 89 days after sowing in 1998, whereas effects induced by WCS358r were detectable for 12 (1997) or 40 (1998) days. None of the strains affected the metabolic activity of the soil microbial population (substrate-induced respiration), soil nitrification potential, cellulose decomposition, plant height, or plant yield. The results indicate that application of GMMs engineered to have improved antifungal activity can exert nontarget effects on the natural fungal microflora.

Antifungal Agents↗

Growth performance and shedding of some pathogenic bacteria in feedlot cattle treated with different growth-promoting agents.

Eighty steers with a mean body weight of 319 kg were used in a study to evaluate the effect of a growth-promoting implant (trenbolone acetate plus estradiol benzoate), monensin, and oxytetracycline on the steer performance and shedding of some foodborne pathogens. The steers were allotted to one of eight treatment combinations according to a randomized complete block design with 16 pens of five animals. Rectal fecal samples were collected before treatment commenced and over a period of more than 24 weeks to study the influence of treatments on the intestinal microbiology of the animals. Results supported the beneficial effect of the hormonal implant on the performance of feedlot steers (average daily gain, feed efficiency, and fat thickness), on carcass characteristics (hot carcass weight, lean yield), and economic value of the carcasses (P < 0.01). The levels of Escherichia coli in feces were not affected by treatments but remained high throughout the study period. Antibiotic-resistant isolates of E. coli were more frequently found as the study progressed but were not associated with any specific treatment. Also independently of treatment, we observed a reduction over time in the shedding of Campylobacter and Yersinia during the feeding period, whereas the shedding of Enterococcus was increased. The results of this study confirmed the beneficial economic effect of growth-promoting agents in beef production and showed that the agents tested did not specifically affect the overall microbial evolution of the animal gut. However, the study also showed, independently of the growth promoter used, the shedding of Campylobacter, Yersinia, and antibiotic-resistant E. coli in the feedlot environment. These bacteria also may be found in the colonic tissue of steers at slaughter and might be a source of carcasses contamination.

Anabolic Agents↗

Usefulness of aerobic microbial culture and cytologic evaluation of corneal specimens in the diagnosis of infectious ulcerative keratitis in animals.

OBJECTIVE: To determine the diagnostic value of aerobic microbial culture and cytologic evaluation of corneal specimens in the diagnosis of infectious ulcerative keratitis (IUK). DESIGN: Prospective study. ANIMALS: 48 animals (26 dogs, 13 horses, 7 cats, 1 bird, and 1 llama) with corneal ulcers. PROCEDURE: Scrapings from corneal ulcers were examined cytologically. Corneal swab specimens were submitted for microbial culture. Animals were grouped according to whether they had been receiving antimicrobials at the time of admission. RESULTS: Of the 38 animals receiving antimicrobials, 19 had positive results for IUK on cytologic evaluation, 20 on microbial culture, and 26 on cytologic evaluation, microbial culture, or both. Of the 10 animals not receiving antimicrobials at the time of admission, 7 had positive results for IUK on cytologic evaluation, and 9 had positive results on microbial culture. In this group of 10 animals, additional animals with IUK were not identified on the basis of cytologic evaluation alone. When all 48 animals were considered irrespective of antimicrobial treatment, 26 and 29 had positive results for IUK on cytologic evaluation and microbial culture, respectively, whereas IUK was confirmed in 35 animals on the basis of cytologic evaluation, microbial culture results, or both. CONCLUSIONS AND CLINICAL RELEVANCE: Microbial culture and cytologic evaluation of corneal specimens maximizes identification of IUK, especially in animals receiving antimicrobial treatment. Because of serious consequences of untreated IUK, we recommend that both diagnostic tests be used to tailor treatment and reduce risk of vision impairment in animals.

Animal Diseases↗