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PubMed2005

Establishment of minimum operational parameters for a high-volume static chamber steam pasteurization system (SPS 400-SC) for beef carcasses to support HACCP programs.

A static chamber steam pasteurization unit (SPS 400-SC()) was installed in a high-volume commercial beef slaughter facility. The SPS 400-SC consists of a three-phase carcass treatment cycle of water removal, steam pasteurization, and water chilling. Seven chamber temperatures (71.1, 73.9, 76.7, 79.4, 82.2, 85.0, and 87.8 degrees C) were evaluated at the midline area of pre-rigor beef carcasses. For each temperature evaluated, 20 carcass sides were randomly selected and aseptically sampled by tissue excision immediately before and after steam pasteurization to determine total aerobic bacteria, Enterobacteriaceae, generic E. coli, and total coliform populations. The 87.8 and 85.0 degrees C treatment temperatures were highly effective at reducing total aerobic bacterial populations, with log(10) reductions of 1.4 and 1.5 CFU/cm(2), respectively, from pretreatment mean population levels of 1.7 and 1.9 log10 CFU/cm(2). These temperatures also reduced Enterobacteriaceae, total coliforms, and generic E. coli to undetectable levels (<0.4 CFU/cm(2)) on all carcasses sampled. Treatment at 82.2 was marginally effective at reducing bacterial populations, while 71.1, 73.9, 76.7, and 79.4 degrees C treatments were ineffective at reducing microbial populations. In a Hazard Analysis Critical Control Points (HACCP)-based system employing steam pasteurization of carcasses as a critical control point, a critical limit of 85.0 degrees C as a minimum chamber temperature should be established, with a targeted operating temperature of 87.8 degrees C providing optimum antimicrobial activity.

Abattoirs↗
PubMed2003

Phylogenetic analysis of anaerobic psychrophilic enrichment cultures obtained from a greenland glacier ice core.

The examination of microorganisms in glacial ice cores allows the phylogenetic relationships of organisms frozen for thousands of years to be compared with those of current isolates. We developed a method for aseptically sampling a sediment-containing portion of a Greenland ice core that had remained at -9 degrees C for over 100,000 years. Epifluorescence microscopy and flow cytometry results showed that the ice sample contained over 6 x 10(7) cells/ml. Anaerobic enrichment cultures inoculated with melted ice were grown and maintained at -2 degrees C. Genomic DNA extracted from these enrichments was used for the PCR amplification of 16S rRNA genes with bacterial and archaeal primers and the preparation of clone libraries. Approximately 60 bacterial inserts were screened by restriction endonuclease analysis and grouped into 27 unique restriction fragment length polymorphism types, and 24 representative sequences were compared phylogenetically. Diverse sequences representing major phylogenetic groups including alpha, beta, and gamma Proteobacteria as well as relatives of the Thermus, Bacteroides, Eubacterium, and Clostridium groups were found. Sixteen clone sequences were closely related to those from known organisms, with four possibly representing new species. Seven sequences may reflect new genera and were most closely related to sequences obtained only by PCR amplification. One sequence was over 12% distant from its closest relative and may represent a novel order or family. These results show that phylogenetically diverse microorganisms have remained viable within the Greenland ice core for at least 100,000 years.

Anaerobiosis↗
PubMed1964

Pure Culture Fermentation of Brined Cucumbers.

The relative abilities of Pediococcus cerevisiae, Lactobacillus plantarum, L. brevis, and several other species of lactic acid bacteria to grow and produce acid in brined cucumbers were evaluated in pure culture fermentations. Such fermentations were made possibly by the use of two techniques, gamma radiation (0.83 to 1.00 Mrad) and hot-water blanching (66 to 80 C for 5 min), designed first to rid the cucumbers of naturally occurring, interfering, and competitive microbial groups prior to brining, followed by inoculation with the desired lactic acid bacteria. Of the nine species tested, strains of the three common to cucumber fermentations, P. cerevisiae, L. plantarum, and L. brevis, grew to the highest populations, and produced the highest levels of brine acidity and the lowest pH values in fermentations at 5.4 to 5.6% NaCl by weight; also, their sequence of active development in fermentations, with the use of a three-species mixture for inoculation, was in the species order just named. This sequence of occurrence was similar to that estimated by others for natural fermentations. The rates of growth and acid production in fermentations with a mixture of P. cerevisiae, L. plantarum, and L. brevis increased as the incubation temperature was increased from 21 to 27 to 32 C; however, the maximal populations and acidities attained were essentially the same for fermentations at each temperature. Further, these same three species were found to be the most salt tolerant of those tested; their upper limit for appreciable growth and measurable acid production was about 8% salt, whereas thermophilic species such as L. thermophilus, L. lactis, L. helveticus, L. fermenti, and L. delbrueckii exhibited a much lower salt tolerance, ranging from about 2.5 to 4.0%. However, certain strains of L. delbrueckii grew very rapidly in cucumbers brined at 2.5 to 3.0% salt, and produced sufficient acid in about 30 hr at 48 C to reduce the brine pH from above 7.0 to below 4.0. An inexpensive, pure culture fermentor which was suitable for gamma radiation, resistant to salt and acid, and which permitted repeated aseptic sampling of the fermenting brine, is illustrated and the specifications are given.

Journal Article↗
PubMed2003

Immunocompetence of juvenile chinook salmon against Listonella anguillarum following dietary exposure to polycyclic aromatic hydrocarbons.

Juvenile chinook salmon (Oncorhynchus tschawytscha) were fed a mixture of 14 polycyclic aromatic hydrocarbon (PAH) compounds that reflected the PAH composition of salmon stomach contents in an urban estuary of Puget Sound, Washington (USA). Following a 28-d dietary exposure, a standardized Listonella anguillarum challenge model was used to determine whether PAH exposure (16, 64, and 252 mg/kg wet wt feed) causes reduced disease resistance under the conditions examined in this study. To assess innate immunity, five replicate groups of fish per dose were acclimated for one week, exposed to a lethal concentration 60 of bacteria, and monitored for 14 d. In a parallel experiment, the effects of PAH exposure on the acquired immune response were examined by immersion vaccinating fish against L. anguillarum and allowing specific immunity to develop for three weeks prior to challenge. All mortalities were aseptically sampled to confirm L. anguillarum infections. No significant differences in fish length, weight, or coefficient of condition were observed. These controlled laboratory experiments suggest that dietary exposures to an environmentally relevant mixture of PAH compounds do not alter the immunocompetence or growth of juvenile chinook salmon.

Administration, Oral↗
PubMed2002

Microbiological quality and somatic cell count of ewe milk with special reference to staphylococci.

A total 1502 useful half udders of 762 Churra ewes from eight herds were aseptically sampled in midlactation to study both the bacteriological isolates and the SCC of milk. Corynebacteria, enterococci, micrococci, staphylococci, and streptococci represented 11.2, 2.9, 1.4, 78.9, and 3.1% of all isolates, respectively. Within staphylococci, novobiocin-sensitive species (71.1%) were much more frequently isolated than novobiocin-resistant ones (7.8%). Staphylococcus epidermidis was the most prevalent species (53.2% of the isolates). Log SCC of uninfected half udder milk was 4.86. Isolates of novobiocin-resistant coagulase-negative staphylococci, micrococci, and corynebacteria were associated to low values of log SCC (4.85 to 5.20). In contrast, infection by novobiocin-sensitive coagulase-negative staphylococci, streptococci, and enterococci organisms was related to a sharp inflammatory response with log SCC means between 5.92 and 6.32. The species that showed the highest log SCC were Pasteurella haemolytica (7.62), Streptococcus agalactiae (7.28), and Staphylococcus aureus (6.68). High prevalence of infections by novobiocin-sensitive staphylococci together with high SCC related to such infections show a relevant role of these organisms in ewe mastitis. Consequently, implementation of staphylococcal mastitis control programs would be of great interest in dairy ewe herds to improve microbiological and hygienic quality of milk.

Animals↗
PubMed1985

Effects of sorbic acid feed fungistat on the intestinal microflora of floor-reared broiler chickens.

The intestinal microflora of a group of broilers fed a diet containing .04% sorbic acid from Days 1 through 49 of their lives was compared with broilers given a diet without the fungistat. Four broilers from each group were killed at 7, 21, 35, and 49 days, and intestinal sections of the duodenum, the lower small intestine, and both ceca were removed aseptically. Samples of the contents of the intestinal sections were analyzed microbiologically. Groups of microorganisms analyzed included total aerobes, total anaerobes, coliforms, streptococci, lactobacilli, clostridia, Bacteroides, Bifidobacterium, yeasts, and molds. Generally, there was a large variation in viable counts of microorganisms for birds of the same treatment. Of the nine groups of microorganisms analyzed the highest counts obtained (greater than 10(6)/g) were for total aerobes, total anaerobes, lactobacilli, and Bifidobacterium. Coliforms, streptococci, and clostridia showed intermediate counts (greater than 10(3)/g), while yeasts and molds and Bacteroides showed the lowest counts (greater than 10(2)/g). Total microbial numbers, in most cases and irrespective of sorbic acid treatment, were highest in the ceca, lower in the small intestine, and still lower in the duodenum. There were no major trends of microbial population changes observed in a given treatment, intestinal location, or group of organisms with age and time on feed. Inclusion of sorbic acid in the feed did not influence total aerobes, total anaerobes, lactobacilli, streptococci, Bifidobacterium, and clostridia. However, broilers on sorbic acid-containing feed had, at 49 days of age, lower coliform counts in the duodenum, lower yeast and mold counts in the ceca, and higher Bacteroides counts in the ceca.

Animal Feed↗
PubMed1999

Identification and control of processing variables that affect the quality and safety of fluid milk.

The objective of this study was to increase quality and safety of fluid milk by eliminating postpasteurization contamination as measured by extended shelf life. Milk shelf life was defined as the number of days for standard plate count to reach 20,000 CFU/ml in milk stored at 7 degrees C. Sequential analysis of the fluid milk processing system indicated filling machine and pasteurizer were significant sources of postpasteurization contamination. Aseptically sampled milk from the pasteurizer outlet indicated a maximum shelf life of more than 30 days could be achieved. The pasteurizer can be a source of contamination when inadequately cleaned or maintained. The filling machine was a significant source of contamination. Shelf life of milk in 236-ml containers was reduced 20 days compared with milk sampled before the filling machine. Carton-forming mandrels, filling heads, and airborne microorganisms were sources of contamination within the filling machine. Eliminating sources of postpasteurization contamination and proper cleaning followed by sanitizing with chlorine significantly increased milk shelf life in paperboard containers to 20.4 days from an initial shelf life of 9 days. Changing the sanitizing agent to peroxyacetic acid significantly increased milk shelf life to 33.9 days. Enclosing the filling chamber and adding sterile laminar flowing air significantly improved microbiological quality of air inside the chamber and reduced variance among milk shelf life samples.

Animals↗
PubMed1999

[Evaluation of the California mastitis test (CMT) reaction in goat milk and its interpretation].

For evaluation of different factors regarding CMT results 65 goats were used. Age, milking hygiene and technique of the animals were recorded and the CMT results were evaluated in relation to those parameters. Correlations between CMT results and age as well milking hygiene and -technique respectively were found. Furtheron the CMT was evaluated as indicator for mastitis diagnosis. Therefore clinical examination of the udder, bacteriological examination of milk samples (aseptically collected) and the determination of the somatic cell count were carried out. The results showed that CMT is not specific for infected udder halves. As important udder pathogens staphylococci were found, S. aureus and CNS at the same level. This investigation has shown that CMT can be used as additional diagnostic tool concerning goat mastitis, but it should not be overestimated because of different factors which influence the cell count. For the control of udder health additional diagnostic measures are of utmost necessity.

Animal Husbandry↗
PubMed2003

Influences of dibutyryl cyclic adenosine monophosphate and forskolin on human sperm motility in vitro.

AIM: To study the influences of dibutyryl cyclic adenosine monophosphate (dbcAMP) and forskolin on human sperm motility in vitro. METHODS: Semen samples, aseptically obtained by masturbation and prepared by swim-up technique from 20 fertile men, were incubated with different concentrations of dbcAMP and forskolin at 37 deg. Measurements were carried out after 10 min, 20 min, 30 min and 60 min incubation. Motility parameters were estimated by using an automatic analyzing system. RESULTS: Treatment with dbcAMP or forskolin resulted in a significant increase in sperm motility and progressive motility. The larger the concentrations of dbcAMP or forskolin, the greater the effect appeared. The straight linear velocity and curvilinear velocity were not affected by both agents. CONCLUSION: dbcAMP and forskolin increase the motility and progressive motility of human sperm in vitro.

Adult↗
PubMed1992

Bacterial patterns found in surgery patients with chronic sinusitis.

Aseptic samples of sinus mucosa or aspirates from maxillary sinuses were taken from 197 patients who underwent sinus surgery and were cultured for aerobic bacteria. Anaerobic cultures were also obtained from 143 of these patients. A total of 236 bacterial isolates from 174 patients were analyzed. No anaerobic organisms were isolated. The most commonly isolated organisms were Staphylococcus species (coagulase-negative), Staphylococcus aureus, gram-negative bacilli, and Streptococcus species. Penicillin G, erythromycin, tetracycline hydrochloride and, to a lesser degree, first-generation cephalosporins were found to be inferior to ciprofloxin, trimethoprim/sulfamethoxazole, and cefuroxime axetil when the susceptibilities of all the bacterial isolates to oral antibiotics used commonly in the empiric therapy of chronic sinusitis were compared.

Bacterial Infections↗
PubMed1977

[Comparative studies of methods for the bacteriological diagnosis of mastitis in cows].

Comparative studies were carried out on some methods and nutrient media used for the isolation and identification of mastitis causative agents. Milk from normal andmastitis-affected udder quarters was aseptically sampled and treated under various conditions. Individual samples were simultaneously seeded in different media. The following conclusions were drawn: 1. Milk samples for the diagnosis of mastitis of a subclinical character can be taken at the beginning and at the end of milking. 2. Direct seedings of milk have equal diagnostic value as those of the sediment. With the former the process of bacteriologic treatment of the samples is shortened in time, and the isolation and differentiation of the mastitis agents is rendered feasible. 3. At the present etiologic structure of the subclinical types of mastitis in this country most suitable has proved the blood agar medium in which almost all mastitis causative agents can be demonstrated. 4. Kartashova's medium and the TKT-medium possess a pronounced selectivity with special reference to streptococci and have higher diagnostic value for farms where streptococcus mastitis is prevailing.

Animals↗
PubMed1980

[Device for sampling and feeding under aseptic conditions].

The scheme and operating principle of a device for sampling and feeding under aseptic conditions are discussed. The device provides sterility of the elements used for sampling and feeding and excludes the contact of the liquids with the environment. The results of the study on the device temperature fields substantiated the sterilization regime using the parameter of the aseptic efficient. The sterilization period is 10 minutes which decreases the steam consumption and increases the production efficiency.

Antisepsis↗
PubMed1992

Detection of enteroviral RNA by polymerase chain reaction in faecal samples from patients with aseptic meningitis.

An assay based on the polymerase chain reaction (PCR) for detection of enteroviral RNA in stool samples was carried out using specimens from 74 patients with aseptic meningitis. The primer pair and probe were derived from the highly conserved 5' non-coding enterovirus genomic region. Enteroviral RNA was detected in faeces of all 36 patients in whom an enterovirus was isolated from stool. The PCR assay yielded positive results in additionally 3/6 cases where enterovirus diagnoses were obtained by virus isolation from cerebrospinal fluid and/or serological tests. Thus, the positive outcome of the PCR assay was 39 (93%) among the 42 patients with enterovirus diagnoses. Furthermore, 7/19 (37%) cases with an etiology that was not established by other means were positive in the test indicating that the PCR assay may give considerable additional etiological information in patients with aseptic meningitis. The limit of RNA detectability in the PCR assay was about 100 TCID50 when highly cytopathogenic enterovirus types (coxsackievirus type B5 and echovirus type 11) were tested. The PCR was negative in all 13 patients with non-enterovirus diagnoses except in one case with a herpes simplex virus type 2 infection. Since enterovirus-specific IgM antibodies could be detected in this case a dual infection seemed probable. All the negative controls, included in the study, were PCR-negative and no contamination was encountered. This study proves the usefulness of the PCR assay for detection of enteroviral RNA in stool samples and suggests that the test may be an alternative to virus isolation for rapid enterovirus diagnosis in patients with aseptic meningitis.

Adolescent↗
PubMed1991

Stability and compatibility of ganciclovir sodium in 5% dextrose injection over 35 days.

The stability and compatibility of ganciclovir sodium in 5% dextrose injection over 35 days were assessed. Nine admixtures of ganciclovir sodium 1, 5, and 10 mg/mL in 5% dextrose injection were aseptically prepared. Immediately thereafter, six samples were aseptically withdrawn from each admixture into sterile collection tubes. Three of the samples were frozen for stability-indicating high-performance liquid chromatographic (HPLC) assay at a later date, and the other three were immediately assessed for pH. Each admixture was also assessed visually for color change, turbidity, gas evolution, and precipitation. The admixtures were stored in the dark at 4-8 degrees C and sampled at 10 and 35 days. There was no significant loss of ganciclovir over the 35-day study period. No admixture at any time contained less than 93.4% or more than 103.7% of its initial ganciclovir concentration. There were no appreciable pH changes, and there was no evidence of visual incompatibility. Ganciclovir sodium 1, 5, and 10 mg/mL in 5% dextrose injection was stable for at least 35 days when stored in the dark at 4-8 degrees C.

Chromatography, High Pressure Liquid↗