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Effects of a twin strain of saccharomyces cerevisiae live cells on mixed ruminal microorganism fermentation in vitro.

This experiment was designed to investigate the effects of different concentrations (0, 0.33, 0.66, 0.99, and 1.32 g/L) of a twin-strain of Saccharomyces cerevisiae live cells on in vitro mixed ruminal microorganism fermentation of corn starch, soluble potato starch, and sudangrass hay (60.5%, DM basis) plus concentrate mixture (39.5%, DM basis). Ruminal fluid was collected from two dairy cows, mixed with phosphate buffer (1:2), and incubated (30 mL) anaerobically at 38 degrees C for 6 and 24 h with or without yeast supplement, using 200 mg (DM basis) of each substrate. Medium pH, ammonia-N, and numbers of protozoa were unaffected (P = 0.38) by yeast cells in all substrates. Molar proportion of acetate was unchanged (P = 0.56) with cornstarch and soluble potato starch, but increased quadratically (P = 0.02) with hay plus concentrate by treatment. Addition of yeast cells caused a linear increase of total VFA (P = 0.008) in all substrates. Excluding the soluble potato starch, supplementation of S. cerevisiae resulted in a quadratic increase of propionate (P = 0.01), with a quadratic decrease (P = 0.04) of acetate:propionate. When soluble potato starch was used as a substrate, a linear increase (P = 0.006) of the molar proportion of propionate and a quadratic decrease (P = 0.007) in acetate:propionate was observed by treatment. Molar proportion of butyrate was unchanged (P = 0.35) with cornstarch and soluble potato starch, whereas it decreased linearly (P = 0.007) with hay plus concentrate by yeast cell supplementation. When cornstarch and soluble potato starch were used as a substrate, minor VFA were decreased (P = 0.05) by treatment. Accumulation of lactate was linearly decreased by treatment (P = 0.007) in all substrates. During incubation with hay plus concentrate, IVDMD was linearly increased (P = 0.006), whereas production of methane (linear; P = 0.02) and accumulation of hydrogen was decreased (quadratic; P = 0.005) by treatment after 24 h. These results showed that a twin strain of S. cerevisiae live cells stimulated in vitro mixed ruminal microorganism fermentation with decreased lactate, and a small decrease of methane and hydrogen with hay plus concentrate.

Ammonia↗

Effects of primary, secondary and tertiary amines on in vitro cellulose digestion and volatile fatty acid production by ruminal microorganisms.

Decanamine hydrochloride (a C10 primary amine), N-methylundecanamine (a C11 secondary amine) and N, N-dimethyldodecanamine (a C12 tertiary amine) were tested for their effects upon in vitro cellulose digestibility by unadapted ruminal microorganisms. The three amines exhibited similar patterns of inhibition of cellulose digestibility in 48-hr in vitro incubations. Cellulose digestion was depressed to 97, 89, 31, 11 and 12% of the control value by 5, 10, 25, 50 and 100 ppm of the amines, respectively. Total volatile fatty acid production was depressed to 89, 85, 61, 40 and 32% of the control value by 5, 10, 25, 50 and 100 ppm of the amines, respectively. Propionate production was inhibited to a greater extent than was acetate production, and acetate to propionate ratios increased in the presence of the amines. The results demonstrate that these primary, secondary and tertiary amines inhibit those microorganisms which digest cellulose in the rumen.

Amines↗

Microorganisms on toothbrushes at day-care centers.

The microflora on 44 toothbrushes at 4 day-care centers in the city of Göteborg have been investigated as a presumptive risk factor for transmission of microorganisms by children. Non-supervised toothbrushing without the use of toothpaste was performed at the day-care centers twice a day. Streptococci, predominantly S. salivarius, S. sanguis, and S. mitis, were the most frequently recorded group of microorganisms and generally constituted the greatest part of the flora (on average, 50%). Beta-hemolytic streptococci were not found in any sample. Haemophilus species were noted in 82% of the samples. H. parainfluenzae being the most frequent, and H. influenzae being identified in only one sample. Anaerobes constituted on average a third of the microflora. Staphylococci were identified in 86% of the samples, S. epidermidis dominating. Fungi including molds were found in 50% of the samples, and from one day-care center large numbers of enteric organisms were identified. Thus this study shows that unsupervised toothbrushing at day-care centers can be questioned, more from a general hygienic point of view than from the risk of transmitting serious pathogens.

Bacteria↗

Mechanisms of halotolerance in microorganisms.

Microorganisms have the ability to adapt to a wide range of NaCl concentrations. In general the NaCl tolerance shown by microbes far exceeds the salt tolerance of any other organism, procryote or eukaryote. There are at least three mechanisms available for adaptation to different salt concentrations. The first would be a passive one in which the cytoplasmic ion content would always equal that in the medium. A second mechanism which is used by many organisms involves concentrating compatible solutes to create an osmotic balance between the cytoplasm and the external environment. The third mechanism involves changing the cell physiology to control the movement of water allowing the cell to exist with an ionically dilute cytoplasm. This article will review the major developments and discuss the implications of increasing knowledge about salt tolerance in microorganisms.

Bacteria↗

Calcium and microorganisms.

This review followed from experiments suggesting that some fungi do not require calcium. It was found that many studies of a calcium requirement in microorganisms had assumed specificity for chelation agents such as EGTA and A23187, which the reagents did not possess. Early studies still cited today often preceded the recognition that microorganisms required manganese and zinc. As a result of both of these misunderstandings, there was rarely any attempt to replace calcium by other important trace elements. In some studies that seem to have been overlooked, the apparent requirement for calcium depended on the growth conditions used. Escherichia coli, Neurospora crassa, and Saccharomyces cerevisiae were then selected for detailed consideration and it is concluded that further experiments are needed before the involvement of calcium is proved.

Bacteria↗

Molecular aspects of pesticide degradation by microorganisms.

Microorganisms are able to degrade a large variety of compounds, including pesticides under laboratory conditions. However, methods have yet to be developed to decontaminate the environment from residues of pesticides. Pesticidal degradative genes in microbes have been found to be located on plasmids, transposons, and/or on chromosomes. Recent studies have provided clues to the evolution of degradative pathways and the organization of catabolic genes, thus making it much easier to develop genetically engineered microbes for the purpose of decontamination. Genetic manipulation offers a way of engineering microorganisms to deal with a pollutant, including pesticides that may be present in the contaminated sites. The simplest approach is to extend the degradative capabilities of existing metabolic pathways within an organism either by introducing additional enzymes from other organisms or by modifying the specificity of the catabolic genes already present. Continuous efforts are required in this direction, and at present several bacteria capable of degrading pesticides have been isolated from the natural environment. Catabolic genes responsible for the degradation of several xenobiotics, including pesticides, have been identified, isolated, and cloned into various other organisms such as Streptomyces, algae, fungi, etc. In addition, recombinant DNA studies have made it possible to develop DNA probes that are being used to identify microbes from diverse environmental communities with an unique ability to degrade pesticides.

Ascomycota↗

High incidence of pathogenic microorganisms in bone allografts explanted in the morgue.

We analyzed the influence of the environment on bacterial contamination in 431 bone allografts from 97 donors (68 multiorgan, grafts taken in the operation theater; 29 morgue donors). From each bone transplant we cultured two aerobic and two anaerobic tissue specimens. The overall contamination rate was 49% (theater 51%, morgue 40%). In grafts explanted in the morgue, we noted more pathogenic microorganisms (60%) than in multiorgan donors, explanted under aseptic surgical conditions (33%). We conclude that the harvesting environment constitutes the major source of pathogenic microorganisms.

Bacteria, Aerobic↗

Effect of sugars and malate on ruminal microorganisms.

The objective of this study was to examine the effects of a commercial feed supplement that contains sugars and malate on lactate fermentation by Selenomonas ruminantium grown in batch culture. Experiments also were conducted to examine the effects of this feed supplement on the mixed ruminal microorganism fermentation of ground corn and soluble starch in the presence and absence of 5 mg/kg of monensin. When S. ruminantium strains HD4 and H18 were incubated in basal medium that contained DL-lactate, some DL-lactate was utilized by both strains after 24 h. In the presence of 1 g/L of sugars plus malate commercial feed supplement, both strains used most of the carbohydrate associated with the feed supplement between 6 and 8 h, and lactate was the main end product. In ground corn fermentations by mixed ruminal microorganisms, 2.25 and 3.25 g/L of sugars plus malate commercial feed supplement increased concentrations of acetate, propionate, and total volatile fatty acids, while 3.25 g/L increased lactate and decreased final pH and butyrate. Fermentation of soluble starch in the presence of both concentrations of sugars plus malate commercial feed supplement increased concentrations of acetate, propionate, and total volatile fatty acids and decreased the acetate:propionate ratio. In the presence of 5 mg/kg of monensin, sugars plus malate treatment increased concentrations of propionate and total volatile fatty acids in ground corn and soluble starch fermentations. Collectively, these results suggest that the sugars plus malate commercial feed supplement stimulates the ruminal fermentation.

Animal Feed↗

Effects of Saccharomyces cerevisiae culture and Saccharomyces cerevisiae live cells on in vitro mixed ruminal microorganism fermentation.

The objective of this study was to examine the effects of a Saccharomyces cerevisiae live cell product and a S. cerevisiae culture product on the in vitro mixed ruminal microorganism fermentation of ground corn, soluble starch, alfalfa hay, and Coastal bermudagrass hay. In the presence of ground corn, neither concentration (0.35 or 0.73 g/L) of S. cerevisiae culture nor live cells had any effect on final pH, H2, CH4, propionate, or butyrate. The S. cerevisiae culture had no effect on acetate, but both concentrations of S. cerevisiae live cells decreased acetate and the acetate:propionate ratio. When soluble starch was the substrate, both concentrations of S. cerevisiae live cells and 0.73 g/L of S. cerevisiae culture decreased the acetate:propionate ratio. Although the treatment effects were not statistically significant, both concentrations of live cells and 0.73 g/L of the culture decreased lactate concentrations compared with the control incubations. When alfalfa hay served as the substrate, neither the S. cerevisiae culture nor the live cells had an effect on propionate, butyrate, or the acetate:propionate ratio. Both concentrations of S. cerevisiae culture decreased the final pH and in vitro dry matter disappearance, and the 0.73 g/L treatment decreased the amount of acetate. However, both treatments of S. cerevisiae live cells increased final pH and decreased acetate and in vitro dry matter disappearance. Neither yeast treatment had much effect on the Coastal bermudagrass hay fermentations. In general, both S. cerevisiae supplements seemed to have similar effects on the mixed ruminal microorganism fermentation.

Acetates↗

Monensin by fat interactions on trans fatty acids in cultures of mixed ruminal microorganisms grown in continuous fermentors fed corn or barley.

In previous studies, monensin (M) and unsaturated plant oils independently increased trans fatty acid concentrations in cultures of mixed ruminal microorganisms. This study was conducted to determine if combining M with plant oil yielded interactions on trans fatty acid concentrations in cultures of mixed ruminal microorganisms or their effects were additive. Four continuous fermentors were fed 14 g of dry feed per day (divided equally between two feedings), consisting of alfalfa hay pellets (30% of DM) and either a high corn (HC) or a high barley (BB) concentrate (70% of DM) in each of two fermentors. Within each grain type, one fermentor was supplemented with M (25 ppm), and the other fermentor was supplemented with 5% soybean oil (SBO) during d 5 to 8. Monensin and SBO were added together in all fermentors during d 9 to 12. Samples were taken at 2 h after the morning feeding on the last day of each period and analyzed for fatty acids by gas-liquid chromatography. A second run of the fermentors followed the same treatment sequence to give additional replication. Average pH across all treatments was 6.15, which was reduced by M but not affected by SBO. Monensin reduced the ratio of acetate to propionate (A:P), which averaged 2.03 across all treatments; fat decreased A:P in cultures not receiving M but increased it in the presence of M. Monensin and SBO altered the concentration of several trans fatty acids, but the only interaction was a grain x M x SBO interaction for trans-10 C18:1. The increase in trans-10 C18:1 by the M and SBO combination exceeded the sum of increases in trans-10 C18:1 for each individual feed additive, but only for KB. For the HC diet, M increased trans-10 C18:1 more than fat alone and more than the M and SBO combination. The results of this study show that M and SBO effects are additive for all trans FA except for trans-10 C18:1. In the case of trans-10 C18:1, M and SBO interacted to give higher trans-10 C18:1 concentrations in ruminal contents than would be expected simply by adding their individual effects, but only for HB. Because some trans fatty acid isomers have been associated with milk fat depression in dairy cows, these results suggest more severe depressions in milk fat content when cows are fed M along with unsaturated plant oils.

Animal Feed↗

Disappearance of docosahexaenoic and eicosapentaenoic acids from cultures of mixed ruminal microorganisms.

Previous studies showed conflicting results regarding the ability of ruminal microorganisms to hydrogenate docosahexaenoic acid (C22:6, DHA) and eicosapentaenoic acid (C20:5, EPA). To determine the disappearance of DHA and EPA from mixed ruminal cultures, 2 ruminal in vitro experiments were conducted using graded levels of DHA and EPA. The first experiment examined DHA added at 0, 5, 10, 15, and 20 mg per culture flask. In the second experiment, EPA was added at 0, 5, 10, and 15 mg per culture flask. Docosahexaenoic acid and EPA were incubated in triplicate in 125-mL flasks, and 5 mL of culture contents was taken at 0, 12, and 24 h for fatty acid analysis by gas liquid chromatography. After 24 h of incubation, 4.1, 4.1, 4.0, and 3.3 mg of DHA disappeared from the 5, 10, 15, and 20 mg of DHA cultures, respectively. In the second experiment, 5, 8.3, and 7.1 mg of EPA disappeared after 24 h of incubation for the 5-, 10-, and 15-mg EPA cultures, respectively. Addition of DHA to cultures increased trans-C18:1 fatty acid accumulation by 105, 91, 82, and 74% for the 5, 10-, 15-, and 20-mg cultures, respectively, compared with control. The addition of EPA increased trans-C18:1 fatty acid accumulation by 56, 64, and 55% for the 5-, 10-, and 15-mg EPA cultures, respectively, compared with control. Addition of DHA and EPA to cultures caused a reduction in C18:1 n-9 and C18:2 n-6 biohydrogenation compared with control. Results from these experiments clearly demonstrate the ability of ruminal microorganism to transform DHA and EPA to other fatty acids causing their disappearance from cultures.

Animals↗

Ameba-associated microorganisms and diagnosis of nosocomial pneumonia.

To elucidate the role of ameba-associated microorganisms (AAMs) as etiologic agents of pneumonia, we screened for Legionella spp., Parachlamydia acanthamoeba, Afipia sp., Bosea spp., Bradyrhizobium spp., Mesorhizobium amorphae, Rasbo bacterium, Azorhizobium caulinodans, Acanthamoeba polyphaga mimivirus, and conventional microorganisms in 210 pneumonia patients in intensive-care units by using culture, polymerase chain reaction, and serologic testing. These resulted in 59 diagnoses in 40 patients. AAMs and non-AAMs were implicated in 10.5% of the patients. The infectious agents were identified in 15 patients: Acanthamoeba polyphaga mimivirus, 8; Legionella pneumophila, 3; L. anisa, 1; Parachlamydia sp., 1; Bosea massiliensis, L. worsleiensis, L. quinlivanii, and L. rubrilucens, 1; and M. amorphae and R. bacterium, 1. A. polyphaga mimivirus was the fourth most common etiologic agent, with a higher seroprevalence than noted in healthy controls. This finding suggested its clinical relevance. Therefore, AAM might cause nosocomial pneumonia and should be suspected when conventional microbiologic results are negative.

Acanthamoeba↗

Comparison of the SimPlate total plate count method with Petrifilm, Redigel, conventional pour-plate methods for enumerating aerobic microorganisms in foods.

The SimPlate Total Plate Count (TPC) method, developed by IDEXX Laboratories, Inc., is designed to determine the most probable number of aerobic microorganisms in foods. The 24-h test was compared to the conventional plate count agar (PCA) method, the Petrifilm Aerobic Count plates, and the Redigel Total Count procedure for enumerating microflora in 751 food samples. Results using the SimPlate TPC method were highly correlated (r > or = 0.96) with results from other test methods. Slopes (0.96-0.97) were not significantly different from 1, and y intercepts (-0.03-0.08) were not different from O. The SimPlate has a high counting range (> 1600 most probable number per single dilution), thus requiring fewer dilutions of samples compared to other methods evaluated. Some foods, e.g., raw liver, wheat flour, and nuts, contain enzymes that gave false-positive reactions on SimPlates. Overall, however, the SimPlate TPC method is a suitable alternative to conventional PCA, Petrifilm, and Redigel methods for estimating populations of mesophilic aerobic microorganisms in a wide range of foods.

Aerobiosis↗

Effect of sampling method on the representative recovery of microorganisms from the surfaces of aquacultured finfish.

The objective of this study was to determine if a gentle rinse procedure was equivalent to the combination of excision and homogenization with a stomacher for the relative removal of various microorganisms from finfish fillets. Fillets of hybrid striped bass and rainbow trout were obtained from local markets and sampled using three methods: rinse (R), excision followed by homogenization in a stomacher (S), and homogenization of fillets following a rinse (RS). Microorganisms were enumerated on selective and nonselective media, and randomly selected colonies from aerobic plate counts were identified using MIDI Sherlock and BIOLOG microbial identification systems. Enrichments and selective media were used for the isolation of Listeria monocytogenes, Salmonella spp., and Yersinia enterocolitica. This study confirms previous reports that stomaching is superior to rinsing for enumerating total microbial populations from fish fillets. Rinsing was more effective for rainbow trout than for striped bass. Sampling method did not affect the relative magnitude of plate counts on media selective for aeromonads, pseudomonads, Shewanella, lactic acid bacteria, enterics, and gram-positive cocci. In the compositional analysis of random isolates, R recovered significantly lower fractions of aeromonads than did S or RS, but sampling method did not affect the percent recovery of lactic acid bacteria, pseudomonads, Shewanella, Moraxellaceae, or Cytophaga/Flavobacterium. However, observations suggest that with increased replication, differences among Moraxellaceae, Pseudomonas, and gram positives might be significant. Only one L. monocytogenes colony was isolated, and no Salmonella or Y. enterocolitica, so the effect of sampling method could not be determined for these organisms. Differences in predominant bacterial populations were seen between fish species.

Animals↗

A new kinetic model for thermal inactivation of microorganisms: development and validation using Escherichia coli O157:H7 as a test organism.

A new kinetic model has been proposed to simulate the nonlinear behavior of survivor curves frequently observed in thermal inactivation of microorganisms. This model incorporates a time component into the first-order inactivation kinetics and is capable of describing the linear, convex, and concave survivor curves. The model was validated using Escherichia coli O157:H7 as a test microorganism. Ground beef (93% lean) samples inoculated to 10(7) to 10(8) CFU/g of meat were subjected to immersion heating at 55, 57.5, 60, 62.5, and 65 degrees C, respectively, in a water bath. All the survivor curves in this study showed upward concavity. Linear and nonlinear regressions were used to fit the survivor curves to the linear first-order inactivation kinetics and the proposed model. Analyses showed that the new kinetic model provides a much better estimate of the thermal inactivation behavior of E. coli O157:H7 in ground beef.

Animals↗

Antimicrobial activity of essential oils from plants against selected pathogenic and saprophytic microorganisms.

The beneficial health effects of extracts from many types of plants that are used as seasoning agents in foods and beverages have been claimed for centuries. The purpose of this study was to examine the effectiveness of selected herb and spice essential oils for control of growth and survival of microorganisms. Inhibition of growth was tested by the paper disc agar diffusion method. Antibiotic susceptibility discs were used as control. Minimum lethal concentration (MLC) was determined by the tube dilution method. Essential oils from anise, angelica, basil, carrot, celery, cardamom, coriander, dill weed, fennel, oregano, parsley, and rosemary were evaluated. Inhibition ranged from complete with oregano to no inhibition with carrot oil for each of the test strains that included: Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O:157:H7, Yersinia enterocolitica, Pseudomonas aeruginosa, Lactobacillus plantarum, Aspergillus niger, Geotrichum, and Rhodotorula. Oregano essential oil showed the greatest inhibition (zone, > or = 70 to 80 mm) (MLC, approximately 8 ppm). Coriander and basil were also highly inhibitory (MLC, approximately 25 to 50 ppm) to E. coli O:157:H7 and to the other bacteria and fungi tested. Anise oil was not particularly inhibitory to bacteria (inhibition zone, approximately 25 mm); however, anise oil was highly inhibitory to molds. Because some of the herbal and spice essential oils are highly inhibitory to selected pathogenic and spoilage microorganisms, they may provide alternatives and supplements to conventional antimicrobial additives in foods.

Apiaceae↗

Standardization of a method to determine the efficacy of sanitizers in inactivating human pathogenic microorganisms on raw fruits and vegetables.

The efficacy of sanitizers in killing human pathogenic microorganisms on a wide range of whole and fresh-cut fruits and vegetables has been studied extensively. Numerous challenge studies to determine the effects of storage conditions on survival and growth of pathogens on raw produce have also been reported. Results of these studies are often difficult to assess because of the lack of sufficient reporting of methods or, comparatively, because of variations in procedures for preparing and applying inocula to produce, conditions for treatment and storage, and procedures for enumerating pathogens. There is a need for a standard method to accurately determine the presence and populations of pathogenic microorganisms on produce. The adoption of standard, well-characterized reference strains would benefit a comparative assessment of a basic method among laboratories. A single protocol will not be suitable for all fruits and vegetables. Modifications of a basic method will be necessary to achieve maximum recovery of pathogens on various types of produce subjected to different sanitizer or storage treatments. This article discusses parameters that must be considered in the course of developing a basic standard method against which these modifications could be made.

Anti-Bacterial Agents↗

Effects of antibacterial dishwashing liquid on foodborne pathogens and competitive microorganisms in kitchen sponges.

In response to increasing concern about home hygiene, the use of antibacterial products to reduce microorganisms in kitchen sponges and cleaning cloths is strongly promoted by some producers of detergent for domestic use. The effects of an antibacterial dishwashing liquid on Escherichia coli, Salmonella Enteritidis, Staphylococcus aureus, and Bacillus cereus were investigated in a modified suspension test and in used sponges with and without food residues under laboratory conditions. A limited study was conducted in households to assess the efficacy of antibacterial dishwashing liquid as used by the consumer. In the suspension tests, S. aureus and B. cereus were shown to be susceptible to low concentrations of antibacterial dishwashing liquid (0.5%), whereas E. coli and Salmonella Enteritidis maintained their initial numbers for at least 24 h at 25 degrees C. At higher concentrations (2 to 4%), all test organisms decreased to below the detection limit after 24 h. Over a 24-h period, the antibacterial dishwashing liquid did not significantly reduce these organisms in used sponges in which food residues were present. The antibacterial product did not reduce the competitive microorganisms either. Similar results were found for sponges involved in daily household use. The results of this study demonstrate that the antibacterial dishwashing liquid was effective in reducing pathogens in the suspension test but not in the used sponges. This finding indicates that to determine the efficacy of antibacterial products, their use in a household setting must be considered.

Bacillus cereus↗