A note on adhesion of bacteria to chicken muscle connective tissue

Laboratory cultures of bacteria were tested for the ability to attach to collagen fibres of intact chicken muscle connective tissue. All salmonellas, fimbriate strains of Escherichia coli and a strain of Campylobacter coli were able to attach to tissue only when suspended in distilled water. Prior immersion of tissue in sterile water for 20 min or extended immersion in these bacterial suspensions was a prerequisite for adhesion. Attachment could be prevented by the addition of physiological levels of sodium chloride to the attachment medium. Furthermore, attached bacteria could be removed by rinsing tissues in saline media.     

Interaction of live and dead Escherichia coli O157:H7 and fluorescent microspheres with lettuce tissue suggests bacterial processes do not mediate adherence

AIMS: The goal of this study was to determine whether any specific bacterial processes (biochemical or genetic) or cell surface moieties were required for the interaction between Escherichia coli O157:H7 and lettuce plant tissue. METHODS AND RESULTS: Escherichia coli O157:H7 and Fluospheres (fluorescent polystyrene microspheres) were used in experiments to investigate interactions with lettuce. Fluospheres were used as they are a non-biological material, of similar size and shape to a bacterial cell, but lack bacterial cell surface moieties and the ability to respond genetically. Live and glutaraldehyde-killed E. coli O157:H7 attached at levels of c. 5.8 log(10) cells per cm(2) following immersion of lettuce pieces into a suspension containing c. 8 log(10) CFU ml(-1). In a separate experiment, numbers of bacteria or Fluospheres associated with lettuce decreased by c. 1.5 log cm(-2) following a 1-min wash. Exposure times of 1 min, 1 h, or 6 h had little effect on the level of attachment for Fluospheres, and live or killed cells of E. coli O157:H7 to lettuce tissue. SIGNIFICANCE: These results indicate that bacterial processes and cell surface moieties are not required for the initial interaction of E. coli O157:H7 to lettuce plant tissue.     

Effects of muscle glycogen and plasma FFA availability on human metabolic responses in cold water.

The purpose of this study was to investigate whether simultaneous alterations in the availability of plasma free fatty acids and muscle glycogen would impair the maintenance of thermal balance during cold water immersion in humans. Eight seminude subjects were immersed on two occasions in 18 degrees C water for 90 min or until rectal temperature (Tre) decreased to 35.5 degrees C. Each immersion followed 2.5 days of a specific dietary and exercise regimen designed to elicit low (LOW) or high glycogen levels (HIGH) in large skeletal muscle groups. Nicotinic acid (1.6 mg/kg) was administered for 2 h before and during immersion to inhibit white adipose tissue lipolysis. Biopsies from the vastus lateralis showed that the glycogen concentration before the immersion was significantly lower in LOW than in HIGH (223 +/- 19 vs. 473 +/- 24 mmol glucose units/kg dry muscle). However, the mean rates of glycogen utilization were not significantly different between trials (LOW 0.62 +/- 0.14 vs. HIGH 0.88 +/- 0.15 mmol glucose units.kg-1.min-1). Nicotinic acid dramatically reduced plasma free fatty acid levels in both trials, averaging 127 +/- 21 mumol/l immediately before the immersion. Cold water immersion did not significantly alter those levels. Plasma glucose levels were significantly reduced after cold water immersion to a similar extent in both trials (18 +/- 4%). Mean respiratory exchange ratio at rest and during immersion was greater in HIGH than LOW, whereas there were no intertrial differences in O2 uptake. The calculated average metabolic heat production during immersion tended to be lower (P = 0.054) in LOW than in HIGH (15.3 +/- 1.9 vs. 17.5 +/- 1.9 kJ/min).(ABSTRACT TRUNCATED AT 250 WORDS)     

Thermal insulation and body temperature wearing a thermal swimsuit during water immersion

This study evaluated the effects of a thermal swimsuit on body temperatures, thermoregulatory responses and thermal insulation during 60 min water immersion at rest. Ten healthy male subjects wearing either thermal swimsuits or normal swimsuits were immersed in water (26 degrees C or 29 degrees C). Esophageal temperature, skin temperatures and oxygen consumption were measured during the experiments. Metabolic heat production was calculated from oxygen consumption. Heat loss from skin to the water was calculated from the metabolic heat production and the change in mean body temperature during water immersion. Total insulation and tissue insulation were estimated by dividing the temperature difference between the esophagus and the water or the esophagus and the skin with heat loss from the skin. Esophageal temperature with a thermal swimsuit was higher than that with a normal swimsuit at the end of immersion in both water temperature conditions (p<0.05). Oxygen consumption, metabolic heat production and heat loss from the skin were less with the thermal swimsuit than with a normal swimsuit in both water temperatures (p<0.05). Total insulation with the thermal swimsuit was higher than that with a normal swimsuit due to insulation of the suit at both water temperatures (p<0.05). Tissue insulation was similar in all four conditions, but significantly higher with the thermal swimsuit in both water temperature conditions (p<0.05), perhaps due to of the attenuation of shivering during immersion with a thermal swimsuit. A thermal swimsuit can increase total insulation and reduce heat loss from the skin. Therefore, subjects with thermal swimsuits can maintain higher body temperatures than with a normal swimsuit and reduce shivering thermo-genesis.     

Comparison of an immunochromatographic method and the TaqMan E. coli O157:H7 assay for detection of Escherichia coli O157:H7 in alfalfa sprout spent irrigation water and in sprouts after blanching

An immunochromatographic-based assay (Quixtrade mark E. coli O157 Sprout Assay) and a polymerase chain reaction (PCR)-based assay (TaqMan E. coli O157:H7 Kit) were used to detect Escherichia coli O157:H7 strain 380-94 in spent irrigation water from alfalfa sprouts grown from artificially contaminated seeds. Ten, 25, 60, or 100 seeds contaminated by immersion for 15 min in a suspension of E. coli O157:H7 at concentrations of 10(6) or 10(8) cfu/ml were mixed with 20 g of non-inoculated seeds in plastic trays for sprouting. The seeds were sprayed with tap water for 15 s every hour and spent irrigation water was collected at intervals and tested. E. coli O157:H7 was detected in non-enriched water by both the TaqMan PCR (30 of 30 samples) and the immunoassay (9 of 24 samples) in water collected 30 h from the start of the sprouting process. However, enrichment of the spent irrigation water in brain heart infusion (BHI) broth at 37 degrees C for 20 h permitted detection of E. coli O157:H7 in water collected 8 h from the start of sprouting using both methods, even in trays containing as few as 10 inoculated seeds. The TaqMan PCR assay was more sensitive (more positive samples were observed earlier in the sprouting process) than the immunoassay; however, the immunoassay was easier to perform and was more rapid. At 72 h after the start of the sprouting process, the sprouts were heated at 100 degrees C for 30 s to determine the effectiveness of blanching for inactivation of E. coli O157:H7. All of the 32 samples tested with the TaqMan assay and 16 of 32 samples tested with the Quixtrade mark assay gave positive results for E. coli O157:H7 after enrichment of the blanched sprouts at 37 degrees C for 24 h. In addition, the organism was detected on Rainbow Agar O157 in 9 of 32 samples after 24 h of enrichment of the blanched sprouts. In conclusion, E. coli O157:H7 was detected in spent irrigation water collected from sprouts grown from artificially contaminated seeds by both the TaqMan and Quixtrade mark assays. The data also revealed that blanching may not be effective to completely inactivate all the E. coli O157:H7 that may be present in sprouts.     

Genetic Transfer of Salmonella typhimurium and Escherichia coli Lipopolysaccharide Antigens to Escherichia coli K-12

Escherichia coli K-12 varkappa971 was crossed with a smooth Salmonella typhimurium donor, HfrK6, which transfers early the ilv-linked rfa region determining lipopolysaccharide (LPS) core structure. Two ilv(+) hybrids differing in their response to the LPS-specific phages FO and C21 were then crossed with S. typhimurium HfrK9, which transfers early the rfb gene cluster determining O repeat unit structure. Most recombinants selected for his(+) (near rfb) were agglutinated by Salmonella factor 4 antiserum. Transfer of an F' factor (FS400) carrying the rfb-his region of S. typhimurium to the same two ilv(+) hybrids gave similar results. LPS extracted from two ilv(+),his(+), factor 4-positive hybrids contained abequose, the immunodominant sugar for factor 4 specificity. By contrast, his(+) hybrids obtained from varkappa971 itself by similar HfrK9 and F'FS400 crosses were not agglutinated by factor 4 antiserum, indicating that the parental E. coli varkappa971 does not have the capacity to attach Salmonella O repeat units to its LPS core. It is concluded that the Salmonella rfb genes are expressed only in E. coli varkappa971 hybrids which have also acquired ilv-linked genes (presumably rfa genes affecting core structure or O-translocase ability, or both) from a S. typhimurium donor. When E. coli varkappa971 was crossed with a smooth E. coli donor, Hfr59, of serotype O8, which transfers his early, most his(+) recombinants were agglutinated by E. coli O8 antiserum and lysed by the O8-specific phage, Omega8. This suggests that, although the parental E. coli K-12 strain varkappa971 cannot attach Salmonella-specific repeat units to its LPS core, it does have the capacity to attach E. coli O8-specific repeat units.     

Interaction of Escherichia coli and soil particles in runoff

A laboratory-scale model system was developed to investigate the transport mechanisms involved in the horizontal movement of bacteria in overland flow across saturated soils. A suspension of Escherichia coli and bromide tracer was added to the model system, and the bromide concentration and number of attached and unattached E. coli cells in the overland flow were measured over time. Analysis of the breakthrough curves indicated that the E. coli and bromide were transported together, presumably by the same mechanism. This implied that the E. coli was transported by advection with the flowing water. Overland-flow transport of E. coli could be significantly reduced if the cells were preattached to large soil particles (> 45 microm). However, when unattached cells were inoculated into the system, the E. coli appeared to attach predominantly to small particles (< 2 microm) and hence remained unattenuated during transport. These results imply that in runoff generated by saturation-excess conditions, bacteria are rapidly transported across the surface and have little opportunity to interact with the soil matrix.     

Adjustments in oxygen transport during head-out immersion in water at different temperatures

Respiratory gas exchange was investigated in human subjects immersed up to the shoulders in water at different temperatures (Tw = 25, 34, and 40 degrees C). Cardiac output (Qc) and pulmonary tissue volume (Vti) were measured by a rebreathing technique with the inert gas Freon 22, and O2 consumption (VO2) was determined by the closed-circuit technique. Arterial blood gases (PaO2, PaCO2) were analyzed by a micromethod, and alveolar gas (PAO2) was analyzed during quiet breathing with a mass spectrometer. The findings were as follows. 1) Immersion in a cold bath had no significant effect on Qc compared with the value measured at Tw = 34 degrees C, whereas immersion in a hot bath led to a considerable increase in Qc. Vti was not affected by immersion at any of the temperatures tested. 2) A large rise in metabolic rate VO2 was only observed at Tw = 25 degrees C (P less than 0.001). 3) Arterial blood gases were not significantly affected by immersion, whatever the water temperature. 4) O2 transport during immersion is affected by two main factors: hydrostatic pressure and temperature. Above neutral temperature, O2 transport is improved because of the marked increase in Qc resulting from the combined actions of hydrostatic counter pressure and body heating. Below neutral temperature, O2 transport is altered; an increase in O2 extraction of the tissue is even calculated.     

Distortion of calculated whole-body hematocrit during lower-body immersion in water

We found a difference between the venous hematocrits of immersed and nonimmersed arms during immersion of the lower body in cold water but not during a comparable exposure to warm water. Fourteen healthy men were exposed to three different experimental conditions: arm immersion, body immersion, and control. The men always sat upright while both upper extremities hung vertically at their sides. During arm immersion, one forearm was completely immersed for 30 min in either cold water (28 degrees C, n = 7) or warm water (38 degrees C, n = 7). This cold-warm water protocol was repeated on separate days for exposure to the remaining conditions of body immersion (immersion of 1 forearm and all tissues below the xiphoid process) and control (no immersion). Blood samples were simultaneously drawn from cannulated veins in both antecubital fossae. Hematocrit difference (Hct diff) was measured by subtracting the nonimmersed forearm's hematocrit (Hct dry) from the immersed forearm's hematocrit (Hct wet). Hct diff was approximately zero when the men were exposed to the control condition and body immersion in warm water. In the remaining conditions, Hct wet dropped below Hct dry (P less than 0.01, 3-way analysis of variance). The decrements of Hct diff showed there were differences between venous hematocrits in immersed and nonimmersed regions of the body, indicating that changes of the whole-body hematocrit cannot be calculated from a large-vessel hematocrit soon after immersing the lower body in cold water.     

Attachment of Salmonella spp. to chicken muscle surfaces.

Immersion of chicken muscle fascia in water or physiological saline caused collagen associated with the connective tissue to expand and form a dense network of fibers on the surface. Similar changes were noted for muscle perimysium. Two test strains of Salmonella spp. attached to the collagen fibers only when muscle was immersed for extended times in water. Bacteria did not attach to the fascia or perimysium of muscle that was transiently immersed in suspensions. The presence of sodium chloride in the suspension media prevented firm attachment, whereas saline rinses removed many attached cells.     

The effect of pyridoxal phosphate on the tryptophanases of five species of Enterobacteriaceae]

Escherichia coli B and E. aurescens, Shigella alkalescens, and Proteus vulgaris et P. morganii tryptophanases (TPases) were studied for the spectral forms of the enzyme. The pH effect on the absorption spectrum and on the enzyme specific activity revealed that the coli group TPases are identical with but differ from Proteus TPases which differ themselves. The coli group TPases attach 4 mol of pyridoxal phosphate (PLP)/mol of enzyme, independently of the pH in the presence of K(plus) ions, and 9 mol of PLP/mol of enzyme must be reduced to achieve complete inactivation. The Proteus TPases attach 4 mol of PLP/mol of enzyme at PH 6.8, and 3 mol of PLP/mol of enzyme at pH 7.8 in K(plus) buffer. In P. morganii, 7 mol of PLP/mol of enzyme must be reduced to inactivate the enzyme, whereas P. vulgaris TPase cannot be completely inactivated by this method. These five TPases attach only 3 mol of PLP/mol of enzyme in a Na(plus) buffer, independently of the pH.     

Blocking adherence of uropathogenic Escherichia coli isolate to HEP-2 cells and bladder of mice in the presence of antibody against p-fimbriae.

Presence of fimbriae on Escherichia coli isolated from the urine of patients with urinary tract infection was related to the ability of the bacteria to attach to human uroepithelial cells. One of the 50 isolates that expresses high MRHA p-fimbriae, selected and antibody against p-fimbriae from it, showed blocking of attachment of bacteria to HEP-2 cell in 1:1024 titer. Also, 1:512 titer of this antiserum to blocking of attachment in bladder tissue of mice is significant.     

Critical water temperature during water immersion at various atmospheric pressures

The present work was undertaken to determine the effect of atmospheric pressure [ranging from a high altitude of 4,300 m above sea level or 0.6 atmospheres absolute (ATA) to depths of 10 m deep or 2 ATA] on the critical water temperature (Tcw), defined as the lowest water temperature a subject can tolerate at rest for 2 h without shivering, of the unprotected subject during water immersion. Nine healthy males wearing only shorts were subjected to immersion to the neck in water at 0.6, 1, and 2 ATA while resting for 2 h. Continuous measurements included esophageal (Tes) and skin (Tsk) temperatures, direct heat loss from the skin (Htissue), and insulation of the tissue (Itissue). The Tcw was significantly higher at 0.6 ATA than 1 and 2 ATA: however, Tcw at 1 ATA was identical to that at 2 ATA. The metabolic heat production remained unchanged among the pressures. During the 2-h immersion in Tcw, Tes was identical among all atmospheric pressures: however, Tsk was significantly higher (P less than 0.05) at 0.6 ATA and was identical between 1 and 2 ATA. The overall mean Itissue was near maximal during immersion in Tcw in each pressure, and no difference was detected among the pressures. However, Itissue at the acral extremities (arm, hand, and foot) decreased significantly at 0.6 ATA, and subsequently heat loss from these parts was increased, which elevated an extremity-to-trunk heat loss ratio to 1.4 at 0.6 ATA from 1.1 at 1 and 2 ATA.(ABSTRACT TRUNCATED AT 250 WORDS)     

Role of atrial natriuretic peptide and urinary cGMP in the natriuretic and diuretic response to central hypervolemia in normal human subjects

The response of plasma atrial natriuretic peptide (ANP) and urinary cGMP excretion to central hypervolemia induced by water immersion was assessed twice in five healthy male subjects, once while immersed in water to the neck for 3 h and again on a control day. Plasma ANP and urinary cGMP were measured by radioimmunoassay. Compared with the control day, overall change in plasma ANP on the immersion day was significant (p less than 0.05). In response to water immersion, plasma ANP increased from a base-line level of 13.2 +/- 3.1 (mean +/- SEM) to 24.2 +/- 5.5 pg/mL by 0.5 h of immersion and was sustained at that level throughout the immersion period. Plasma ANP returned to the base-line level at 1 h postimmersion. Urinary cGMP excretion increased significantly by 1 h of immersion and was sustained at that level throughout water immersion and 1 h postimmersion (p less than 0.05). During water immersion urine flow, urinary sodium and potassium excretion, free water clearance, and osmolar clearance increased while plasma renin activity, serum aldosterone, and blood pressure fell; all changes were significant (p less than 0.05). Creatinine clearance and hematocrit did not show any significant changes. These data suggest that an increase in plasma ANP may contribute to the natriuretic and diuretic response to central hypervolemia, and that the measurement of urinary cGMP may be a valuable marker of ANP biological responsiveness.     

Effect of pressure on thermal insulation in humans wearing wet suits

The present work was undertaken to determine the critical water temperature (Tcw), defined as the lowest water temperature a subject can tolerate at rest for 3 h without shivering, of wet-suited subjects during water immersion at different ambient pressures. Nine healthy males wearing neoprene wet suits (5 mm thick) were subjected to immersion to the neck in water at 1, 2, and 2.5 ATA while resting for 3 h. Continuous measurements of esophageal (T(es)) and skin (Tsk) temperatures and heat loss from the skin (Htissue) and wet suits (Hsuit) were recorded. Insulation of the tissue (Itissue), wet suits (Isuit), and overall total (Itotal) were calculated from the temperature gradient and the heat loss. The Tcw increased curvilinearly as the pressure increased, whereas the metabolic heat production during rest and immersion was identical over the range of pressure tested. During the 3rd h of immersion, Tes was identical under all atmospheric pressures; however, Tsk was significantly higher (P less than 0.05) at 2 and 2.5 ATA compared with 1 ATA. A 42 (P less than 0.001) and 50% (P less than 0.001), reduction in Isuit from the 1 ATA value was detected at 2 and 2.5 ATA, respectively. However, overall mean Itissue was maximal and independent of the pressure during immersion at Tcw. The Itotal was also significantly smaller in 2 and 2.5 ATA compared with 1 ATA. The Itissue provided most insulation in the extremities, such as the hand and foot, and the contribution of Isuit in these body parts was relatively small. On the other hand, Itissue of the trunk areas, such as the chest, back, and thigh, was not high compared with the extremities, and Isuit played a major role in the protection of heat drain from these body parts.     

Ecological relationships between Vibrio cholerae and planktonic crustacean copepods.

Strains of Vibrio cholerae, both O1 and non-O1 serovars, were found to attach to the surfaces of live copepods maintained in natural water samples collected from the Chesapeake Bay and Bangladesh environs. The specificity of attachment of V. cholerae to live copepods was confirmed by scanning electron microscopy, which revealed that the oral region and egg sac were the most heavily colonized areas of the copepods. In addition, survival of V. cholerae in water was extended in the presence of live copepods. Attachment of viable V. cholerae cells to copepods killed by exposure to -60 degrees C was not observed. Furthermore, survival of V. cholerae was not as long in the presence of dead copepods as in the live copepod system. A strain of Vibrio parahaemolyticus was also seen to attach to copepod surfaces without effect on survival of the organism in water. The attachment of vibrios to copepods was concluded to be significant since strains of other bacteria, including Pseudomonas sp. and Escherichia coli, did not adhere to live or dead copepods. Attachment of V. cholerae to live copepods is suggested to be an important factor of the ecology of this species in the aquatic environment, as well as in the epidemiology of cholera, for which V. cholerae serovar O1 is the causative agent.     

Is water immersion useful for analyzing gravity resistance responses in terrestrial plants?

Water immersion has been used as a simulator of microgravity for analyzing gravity responses in semiaquatic plants such as rice. To examine whether or not water immersion for a short experimental period is a useful microgravity simulator even in terrestrial plants, we analyzed effects of water immersion on the cell wall rigidity and metabolisms of its constituents in azuki bean epicotyls. The cell wall rigidity of epicotyls grown underwater was significantly lower than that in the control. Water immersion also caused a decrease in molecular mass of xyloglucans as well as the thinning of the cell wall. Such changes in the mechanical and chemical properties of the cell wall underwater were similar to those observed in microgravity conditions in space. These results suggest that water immersion for a short period is a useful system for analyzing gravity resistance responses even in terrestrial plants.     

The dynamic viscoelasticity and plasticizer leachability of tissue conditioners

doi: 10.1111/j.1741‐2358.2012.00639.x
The dynamic viscoelasticity and plasticizer leachability of tissue conditioners Objectives: Dynamic viscoelasticity is one of the most important characteristics of tissue conditioners. Leaching of plasticizer from tissue conditioners may cause changes in the dynamic viscoelasticity of these materials. The purpose of this study was to determine the dynamic viscoelasticity and plasticizer leachability of commercial tissue conditioners. Materials and methods: Five commercial tissue conditioners were used in this study: COE Comfort (CC), Fit Softer (FS), Hydro‐Cast (HC), Soft Conditioner (SC) and Visco‐Gel (VG). Five specimens of each material were stored in 37°C distilled water. The dynamic viscoelasticity and plasticizer leaching of each specimen were measured at 0, 1, 3, 7 and 14 days after immersion using a dynamic mechanical analyzer and high performance liquid chromatography. Data were analyzed using one‐way anova and Student–Newman–Keuls test (α = 0.05). Results: Significant differences in dynamic viscoelasticity and plasticizer leaching were found among the materials. The dynamic viscoelasticity of all materials increased or decreased significantly with time. Rapid changes in dynamic viscoelasticity were evident on day 1 after water immersion. The material CC recorded the highest level of plasticizer leaching. Conclusion: These results suggest that the dynamic viscoelasticity of commercial tissue conditioners is affected by plasticizer leaching. However, these effects are limited.     

Modification of stress tolerance by dehydration pretreatment in winter rape hypocotyls

Dehydration of isolated rape hypocotyls (Brassica napus L. var. oleifera L. cv. Górczański) to a water saturation deficít of about 60% resulted in increased tolerance to subsequent desiccation and to prolonged immersion in water. Dehydration pretreatment followed by restoration of tissue turgidity, caused a decrease in leakage of electrolytes, ultraviolet-absorbing materials and ninhydrin-reactive substances from the desiccated or water immersed hypocotyl tissue. A hardening effect of tissue dehydration is supposed to be related to an improved stability of cell membranes.     

Enterococci as indicators of Lake Michigan recreational water quality: comparison of two methodologies and their impacts on public health regulatory events

The frequency of poor-water-quality advisories issued in Milwaukee and Racine, Wisconsin, in the absence of identifiable sources of contamination brought into question the reliability of the present indicator organism, Escherichia coli. Enteroccoci have been suggested as an alternative to E. coli for freshwater monitoring due to their direct correlation to swimmer-associated gastroenteritis. The purpose of this research was threefold: (i) to explore enterococci as an alternative to E. coli for monitoring freshwater Lake Michigan beaches, (ii) to evaluate the impact of the two indicators on regulatory decisions, and (iii) to compare membrane filtration m-enterococcus agar with indoxyl-beta-D-glucoside to a chemical substrate technique (Enterolert) for the recovery of enterococci. Recreational water samples from Milwaukee (n = 305) and Racine (n = 153) were analyzed for the enumeration of E. coli and enterococci using IDEXX Colilert-18 and Enterolert. Correlation between the indicators was low (R(2) = 0.60 and 0.69). Based on U.S. Environmental Protection Agency bacterial indicator threshold levels of risk for full body immersion, using enterococci would have resulted in 56 additional unsafe-recreational-water-quality advisories compared to the total from using E. coli and the substrate-based methods. A comparison of the two enterococcal methods (n = 124) yielded similar results (R(2) = 0.62). This was further confounded by the frequent inability to verify enterococci from those wells producing fluorescence by the defined substrate test using conventional microbiological methods. These results suggest that further research is necessary regarding the use of defined substrate technology interchangeably with the U.S. Environmental Protection Agency-approved membrane filtration test for the detection of enterococci from fresh surface water.