Refrigerated femoral venous-venous haemodialysis with coil preservation for rehabilitation of terminal uraemic patients.
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Canine epididymides were excised and immediately stored at 4 degrees C for 48 hr, and the qualities of caudal epididymal sperm after recovery and cryopreservation were evaluated. To confirm the fertility of the cryopreserved caudal epididymal sperm, artificial intrauterine insemination was performed. The sperm motility (61.0%) immediately after recovery from caudal epididymis stored at 4 degrees C for 48 hr was significantly lower than those of sperm stored for 0 and 24 hr (88.6 and 80.7%, respectively), but there was no significant difference after freeze-thawing (0-, 24-, and 48-hr storage groups: 27.9, 24.3, and 28.3%, respectively). The incidence of abnormal sperm immediately after recovery was significantly higher in the 24-hr and 48-hr storage groups (19.3 and 27.7%, respectively) than in the 0-hr storage group (5.6%), and a significant difference was also observed after freeze-thawing. The incidence of immature sperm with cytoplasmic droplets was significantly higher in the 48-hr storage group (18.4%) than in the 0-hr storage group (4.7%), but there was no difference after freeze-thawing. By unilateral intrauterine insemination (2x10(8) sperm), 4 of 5 bitches (80%) conceived. The above findings demonstrated that sperm motility was good even enough the incidence of abnormal sperm was high in canine epididymal sperm that were recovered from the epididymis stored at 4 degrees C for 48 hr and cryopreseved, and that artificial intrauterine insemination resulted in a high conception rate.
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Cells of Lactobacillus acidophilus propagated in peptonized milk nutrient broth maintained at pH 5 were harvested during the late logarithmic (log) phase and at 6 h into the stationary phase of growth. Concentrated cultures were prepared from each age of cells, frozen, and stored at -196 degrees C. The concentrated cultures were assayed for numbers of total and bile-tolerant lactobacilli, beta-galactosidase activity, and ability to assimilate cholesterol before and after storage at -196 degrees C. Nonfermented acidophilus milk, prepared following 28 d of storage at -196 degrees C from each concentrated culture, was tested before and after 0, 7, 14, 21, and 28 d of storage at 7 degrees C. No decrease in numbers of total or bile-tolerant lactobacilli, beta-galactosidase activity, or amount of cholesterol assimilated was observed for any of three strains studied during storage at -196 degrees C. There were no significant differences between the two harvest times during storage at this temperature for any of the three strains. However, when the cells were suspended in milk and stored at 7 degrees C, the numbers of total and bile-tolerant lactobacilli declined over time as did the beta-galactosidase activity and the ability to assimilate cholesterol. The cells from L. acidophilus 223, harvested 6 h into the stationary phase, exhibited significantly less decrease in total numbers during extended storage at 7 degrees C than did the cells harvested in the late logarithmic phase. There was little or no difference in the decline in numbers of total lactobacilli between the two harvest times for strains 606 and 107. All strains decreased significantly in beta-galactosidase activity and the ability to assimilate cholesterol during storage in milk at 7 degrees C with no significant differences between the two harvest times.
The effect storage at 4 degrees C on the viability of Lactobacillus gasseri and its sodium taurocholate-deconjugating and cholesterol-binding abilities as well as desmutagenic activity was investigated. Unfermented milks containing L. gasseri strains SBT0274 and SBT0270 at 10(9) cfu/ml were prepared using 10% skim milk. Total and bile-tolerant lactobacilli for strains SBT0274 and SBT0270 generally decreased after 14 d of storage at 4 degrees C; however, viable cells of these strains were still at 10(8) cfu/ml after 28 d of storage. The amounts of cholic acid released and of cholesterol bound by strains SBT0274 and SBT0270 declined over time, especially at 21 d of storage. Antimutagenic activity of unfermented milk made from both strains was attributed to the bacterial cells, and the activity was stable during storage at 4 degrees C for 28 d.
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Poliovirus survival in live and frozen mussels during storage was assessed by both viral culture and molecular methods. Live New Zealand green-lipped mussels were incubated overnight at 20 degrees C in an aerated tank of filtered seawater seeded with the poliovirus 2 (PV2) vaccine strain. An extraction and concentration method that preserved viral infectivity was used to recover PV2 taken up by the mussels at day 0, at day 2 after storage at 4 degrees C, and at days 7, 14, and 28 after storage at -20 degrees C. This method allowed both culture and molecular analysis to be carried out. Presence of intact PV2 in each batch of mussels was determined by a pan-enterovirus specific reverse transcription-polymerase chain reaction (RT-PCR) and confirmed by dot-blot hybridization. Survival of infectious PV2 was determined by the monolayer plaque assay. After 48 h at 4 degrees C, infectious PV2 levels were 81% of the original level detected in the mussels. Infective virus levels then declined to 66, 53, and 44% after storage at -20 degrees C for 7, 14, and 28 days, respectively. Generic RT-PCR methods were 10 times more sensitive than cell culture techniques for virus detection but did not give information on virus infectivity. The survival of infectious pathogenic viruses in fresh and frozen mussels on storage constitutes a potential health risk and so is a major concern for public health authorities.
Studies were done to determine the survival of salmonellae in orange juice as affected by fortification with calcium. Four brands of commercially pasteurized orange juice fortified with calcium (350 mg/240-ml serving) and nonfortified juice were inoculated separately with three types of inocula: strains of Salmonella Muenchen (inoculum 1), serotypes of human and animal origin (inoculum 2), and isolates from raw produce- and juice-associated outbreaks (inoculum 3). Juice inoculated with populations of 6.6 to 7.0 log10 CFU of Salmonella per ml was held at 4 degrees C for up to 32 days. The number of cells of inoculum 1 that survived in juice fortified with calcium lactate/tricalcium phosphate (CaL/TCP) was significantly lower (P < or = 0.05) (2.80 log10 CFU/ml) than in nonfortified juice (3.50 log10 CFU/ml) after 32 days' storage. Death of salmonellae in inocula 1 and 2 was less in juice fortified with TCP (3.21 and 3.33 log10 CFU/ml, respectively) than in the nonfortified juice (3.75 and 4.15 log10 CFU/ml, respectively). During the 32-day storage period, populations in inocula 1 and 3 showed significantly less inactivation (2.62 and 3.12 log10 CFU/ml, respectively) in juice fortified with calcium citrate (CC) than in nonfortified juice (3.14 and 3.60 log10 CFU/ml, respectively). There were no significant differences in the survival of Salmonella in juice fortified with calcium citrate malate (CCM) and nonfortified juice. Polymerase chain reaction (PCR) typing of randomly selected Salmonella colonies revealed that Salmonella Heidelberg in inoculum 2 and Salmonella Baildon and Salmonella Poona in inoculum 3 were the most prevalent at the end of the 32-day storage period at 4 degrees C, suggesting that serotypes selected for use in inocula differed in tolerance to acidic environments. This study reveals that the form of calcium used to fortify orange juice may affect the survival of Salmonella.