Recommended culture methods for monitoring bacterial contamination of H2O, dialysate and bicarbonate concentrate in dialysis centers in the USA involves culturing these fluids for 48 h at 37 °C. A variety of media and commercial culture methods are accepted for monitoring these fluids. Over a 3 month period a comparison was made between an acceptable culture method, tryptic soy agar (TSA) employing the pour plate (PP) technique at 37 °C for 48 h, and PP cultures on standard methods agar (SMA) and R2A agar, incubated at ambient temperature (23°C) for 48, 72, 168 h. Increases in the colony counts over time occurred for all three fluids. However, counts were greater on SMA and R2A than on TSA. The increases over the standard 48-hour TSA cultures ranged as high as 104 times for 23°C cultures at 7 days of incubation. Endotoxin levels even in the most contaminated samples were found to be below the acceptable 5 EU/ml recommended for reprocessor water. Bacterial colonies that appeared at 48, 72 and 168 h were isolated and identified. Pseudomonas, Moraxella, Acinetobacter and CDC group VI C-2 were among some of the common bacteria isolated. This study indicates that the media utilized, the time and temperature of incubation may result in a significant underestimation of the bacterial population of water and dialysis fluids, thus potentially placing the patient at a higher risk.
Many dialysis centers depend on clinical laboratories or a commercially available dip culture to determine the contamination levels in water and dialysate. To determine whether these standard clinical culture procedures adequately quantitate bacterial contamination in hemodialysis center water and dialysate, test results of two routine clinical media was compared, trypticase soy agar (TSA) and plate count agar (PCA), with those of nutrient-poor R2A medium. Dialysate samples demonstrated significant differences in media, the temperature of incubation, and plating techniques (pour plate versus spread plates). Purified water for dialysis demonstrated significant differences only for media; however, temperature was an important variable. Selective growth on R2A agar of some water- and dialysate-contaminating species was studied by velvet disk and loop transfer of colonies. A strong selectivity for water-borne bacteria was demonstrated by R2A agar; the bacteria that did not grow on TSA and PCA have been identified.
1. Gene frequencies of esterase D, glyoxalase I and uridine monophosphate kinase in Alaskan populations were determined. 2. Improved methods for demonstrating phenotypes of glyoxalase I and uridine monophosphate kinase are presented. 3. Rare variants of adenylate kinase (AK1) and diaphorase (DIA) were found. One of the AK1 variants is new. 4. Gene frequencies were notably diverse within major ethnic groups. This variability was consistent with a population structure composed of small groups that were relatively isolated from one another.
The purified water supplies and randomly selected dialysates of 51 chronic and acute dialysis centers in the central United States were surveyed to assess the relative risks to dialysis patients from microbial and endotoxin contamination. A culture medium more sensitive than those generally employed in routine quality assurance assays was used for recovery of bacteria from water. With this medium, 35.3% of the water samples and 19% of the dialysate samples were out of compliance with the Association for the Advancement of Medical Instrumentation (AAMI) standards: 200 and 2,000 colony forming units (CFU)/ml, respectively. There was no correlation observed between the type of water purification system or the frequency of disinfection of the system and the bacterial and endotoxin contamination levels. There was also no correlation found between the bacterial and fungal CFU per ml and the endotoxin concentration per ml (EU/ ml). It is recommended that more sensitive culturing methods be used to provide adequate bacterial monitoring of dialysate center water supplies. Dialysis centers should monitor endotoxin in dialysate on a regular schedule and immediately after any endotoxemic‐like patient reactions. Yeast and fungi were observed in 10% and 64% of the water systems, respectively. Dialysate was contaminated by yeast and fungi in 30% and 70% of the centers, respectively. The concentrations of these microbes in both fluids were much lower than bacteria. However, they were observed often enough to warrant further investigation of their impact on the well‐being of dialysis patients.
Biomagnification of methylmercury in aquatic systems can cause elevated tissue mercury (Hg) and physiological stress in top predators. Mercury is known to affect stress hormone levels in mammals, birds and fish. In this study, the effects of maternally-transferred methylmercury on the stress physiology of Northern Water Snake (Nerodia sipedon) neonates were tested. Gravid females were dosed via force-fed capsules during late gestation with 0, 0.01, or 10 µg methylmercury per gram of body mass. Plasma corticosterone levels and leukocyte differentials were analyzed in baseline and confinement-stressed neonates from all dose levels. Neither Hg nor confinement stress had a significant effect on leukocyte differentials nor was Hg related to corticosterone levels. However, stress group neonates showed lower heterophil/lymphocyte ratios and this study was the first to show that neonate N. sipedon can upregulate CORT in response to stress. These results indicate that N. sipedon may be somewhat tolerant to Hg contamination.
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