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PubMed2002

Comparison of eggshell surface microbial populations for in-line and off-line commercial egg processing facilities.

The objective of this project was to evaluate the aerobic plate counts (APC) of eggshells at in-line and off-line egg processing facilities at selected sites, throughout the processing procedure. Samples were collected from four sites in the processing plant and five time periods during the daily processing shift. Site 1 was from the conveyor system before the eggs passed through the washing system. Site 2 was after detergent wash but before sanitizer application. Site 3 was immediately after sanitizer treatment. Site 4 was immediately before packaging. Samples were collected from the sites at five equally spaced intervals beginning 15 min after the processing shift began and ending 15 min before the processing shift ended. At each sampling time, eggs were aseptically collected from each site and placed into sterile plastic bags containing 50 mL of PBS that was serially diluted immediately. The dilutions were plated on APC agar within 8 h of collection and were incubated at 37 C for 48 h. APC counts of in-line and off-line eggs were compared within time periods across sites. As the processing shift progressed, off-line APC counts were significantly higher than in-line counts at Site 1. At Site 2, off-line APC counts were significantly higher than in-line counts for Periods 2 through 5. At Site 3, off-line APC counts were significantly higher than in-line counts for Periods 2 through 5. Site 4 off-line counts were significantly higher than in-line counts at all time periods.

Animals↗
PubMed1993

Postnatal changes in selected bacterial groups of the pig colonic microflora.

The importance of the colonic microflora in health and nutrition is well known, but how they colonize and become established in the colon is not well understood. We therefore characterized the quantitative and qualitative changes of the colonic microflora during the first 120 days of postnatal development. Unlike previous studies, changes were defined for individual pigs using in situ samples collected anaerobically and aseptically from the distal colon. Although the colons were sterile at birth, they were rapidly colonized, and within 12 h bacterial densities had stabilized at 10(-9)-10(10) bacteria/g colonic content. Facultative anaerobes, notably coliforms, initially dominated the microflora, but were supplanted within 48 h after birth by obligate anaerobes, which constituted greater than 90% of the microflora thereafter. Bacteroides spp., the predominant anaerobes in the adult colon, did not markedly increase in abundance until after weaning and were still increasing by postnatal day 120. Shifts in the relative abundances of different bacterial populations throughout the first 120 days after birth confirm previous reports that the establishment of the adult colonic microflora is a gradual, sequential process, and highlight the need to focus research on anaerobic groups.

Aging↗
PubMed1997

Effect of dietary fiber on bowel mucosal integrity and bacterial translocation in burned rats.

The response of the bowel mucosa to enteral formula supplemented with dietary fiber was examined in rats with 30% full-thickness burns. The rats were fed a standard enteral formula without fiber or with one of two types of fiber (insoluble soy fiber or soluble guar gum fiber). Seventy-two hours after burn injury, the mesenteric lymph nodes were excised aseptically for bacterial culturing. Samples of the jejunum, ileum and cecum were also collected for histological examination. There were significantly fewer bacterial colonies in the lymph node cultures from rats given soy fiber compared to those from rats fed no fiber. In rats given soy fiber, the integrity of the bowel mucosa was maintained in the jejunum, ileum and cecum. In rats given guar gum fiber, however, the repair of mucosal erosions was observed in the jejunum and ileum as well as flattening of the cecal mucosa. These findings indicate that soy fiber is superior to guar gum fiber for maintaining bowel mucosal integrity and preventing bacterial translocation in burned rats receiving enteral feeding.

Animals↗
PubMed1980

Growth characteristics of human diploid fibroblasts in packed beds of glass beads.

Measurement of cell numbers during growth on glass beads is difficult because of the problem of taking a representative sample of the beads aseptically. Growth is therefore estimated by indirect means, predominantly the rate of glucose utilisation. The relationship between rates of glucose utilisation and cell growth is discussed.

Cell Count↗
PubMed1993

Cluster of Enterobacter cloacae pseudobacteremias associated with use of an agar slant blood culturing system.

From 1 February through 12 October 1990, 27 blood cultures processed at Shiprock Hospital were positive for Enterobacter cloacae; only 3 had been reported in the preceding 12 months. Twenty (74%) of the cultures were obtained from patients without clinical evidence of gram-negative septicemia. The increase in E. cloacae-positive blood cultures was temporally associated with the introduction of a new blood culturing system. To evaluate potential risk factors for an E. cloacae-positive blood culture (case-culture), we conducted a case-control study. Case-cultures were compared with 81 randomly selected cultures that were processed during the epidemic period and that were not positive for E. cloacae (controls). Because several factors suggested the possibility of pseudoinfection, we limited our analysis to the 20 blood cultures that appeared to be contaminants. Blood samples received in the laboratory during the midnight shift (5 of 20 [25%] versus 5 of 81 [6%]; odds ratio, 5.1; 95% confidence intervals, 1.01 to 24.6; P = 0.02) or present in the incubator with other E. cloacae-positive samples (17 of 20 [85%] versus 29 of 81 [36%]; odds ratio, 10.2, 95% confidence interval, 2.6 to 57.3; P < 0.001) were at increased risk for contamination. During mock experiments of the procedures for processing blood samples for culture, several breaks in aseptic technique and leakage from the blood culturing system were observed. Cultures of samples obtained from several environmental sites in the laboratory and the hand washings of two laboratory technicians grew E. cloacae. Plasmid and restriction enzyme analyses of E. cloacae isolates recovered from the patients' blood cultures, the two technicians' hand washings, and environmental sites in the laboratory indicated that all had identical plasmid profiles. Our findings suggest that the breaks in aseptic technique and the environmental contamination that occurred in association with the use of the new blood culturing system resulted in contamination of the blood cultures. This outbreak highlights the importance of routine environmental cleaning, periodic quality control assessments, and adherence to aseptic practices in clinical laboratories, particularly when new methods or equipment are introduced and/or new personnel are hired.

Agar↗
PubMed1999

Evaluation of combining upper respiratory tract swab samples with cerebrospinal fluid examination for the diagnosis of enteroviral meningitis in children.

In a prospective comparative study, the use of combined analysis of upper respiratory tract swab samples and cerebrospinal fluid (CSF) samples was assessed to improve the detection rate of enteroviral meningitis in children. An enterovirus was detected in 32% of patients with aseptic meningitis when testing CSF samples alone compared with 71.5% when combining CSF and respiratory tract findings. An enterovirus was detected in 17% of respiratory tract samples in an age- and sex-matched control group without meningitis. Thus, combining the examination of upper respiratory tract with CSF findings may improve the detection rate of enteroviral meningitis. Upper respiratory tract samples should be included in the diagnosis scheme to differentiate benign enteroviral meningitis from other life-threatening infections of the central nervous system.

Adolescent↗
PubMed2001

Investigation of bacteremia after orthodontic banding and debanding following chlorhexidine mouth wash application.

This study investigates the prevalence of bacteremia after orthodontic banding and debanding, following the application of a 0.2% chlorhexidine gluconate mouthwash. The banding and debanding groups were each composed of 40 young adult patients. In the banding group, patients were asked to rinse their mouth with chlorhexidine gluconate for 60 seconds just prior to fitting of the bands. In the debanding group, they were asked to use the mouthwash immediately before removal of bands and brackets. In both groups pre- and post-treatment blood samples were obtained with a strict aseptic technique. In the banding group, no bacteremia was detected in the pretreatment sample and 2.5% post-treatment bacteremia was detected in the post-treatment sample. In the debanding group, 2.5% bacteremia was found in both the pre- and post-treatment samples. The prevalence of post-treatment bacteremia found in the present study were compared with the findings of 2 preliminary studies in which the prevalence of bacteremia had been investigated after banding and debanding without a prior application of chlorhexidine mouthwash. The application of chlorhexidine mouthwash resulted in a decrease in the prevalence of bacteremia after banding and debanding, but the decrease was not statistically significant.

Adolescent↗
PubMedDate not supplied

Detection of enteroviruses in cases of neurological disorders in the State of Pará, Brazil.

Eighty-one cerebrospinal fluid (CSF) samples mainly from cases of aseptic meningitis and motor deficiency syndrome were sent to the Virology Section of Evandro Chagas Institute, Belém Pará, in the period of January 1995 to January 1996 in order to isolate viruses. All samples were inoculated onto HEp-2 cell culture and newborn mice, with negative results. The probability of isolating viruses by these methods is reduced because of the low concentration of viral particles in these specimens. In order to obtain more information about the etiology of these cases, a group of 23 samples were selected to be tested by a more sensitive technique than the virus isolation - the reverse transcription polymerase chain reaction (RT-PCR). Specific primers directed to conserved regions in the enterovirus genome were used, considering that this group of viruses is frequently associated with these neurological disorder. The age of the patients ranged from 1 to 55 years and nearly all of them lived in Belém, State of Pará, North of Brazil. Of 15 samples analyzed by RT PCR nine (60%) were positive; of these, 6 (66.6%) had motor deficiency and 3 (33.3%) developed aseptic meningitis. These results show that it is important to investigate enterovirus as cause of these syndromes.

Adolescent↗
PubMed2001

Detection of Campylobacter spp. in ceca and crops with and without enrichment.

The purpose of this experiment was to determine how sampling method (direct plating or enrichment) affected the rate of Campylobacter spp. isolation from crop and cecal samples. In four separate trials, 32 New York-dressed broiler carcasses were obtained from commercial plants (n = 128). Crops and ceca were removed aseptically, direct plated, and enriched. Samples were direct-plated on Campy-Cefex plates that were incubated at 42 C for 36 to 48 h under a microaerobic atmosphere (5% O2, 10% CO2, 85% N2). After direct plating, samples were enriched in Bolton broth at 37 C for 4 h and 42 C for 20 h under a microaerobic atmosphere before plating onto Campy-Cefex plates. Campylobacter spp. was detected in 95.3% of direct-plated crop samples and 99.2% of enriched crop samples. Campylobacter spp. was detected in 100% of direct-plated cecal samples and 63.3% of enriched cecal samples. All 128 crop and cecal samples were positive for the organism by one or both methods. Mean counts of Campylobacter spp. were 3.6 log10 cfu/g of crop sample and 6.8 log10 cfu/g of cecal sample. For these two sample types, both of which tend to be contaminated with many viable cells, direct plating is sufficient for isolation of Campylobacter. Direct plating also provides an estimate of contamination level. Enrichment of cecal samples resulted in a decreased rate of detection and did not allow estimation of numbers of Campylobacter. The large numbers of non-Campylobacter species that inhabit the intestinal tract may out-compete Campylobacter during enrichment, confounding detection.

Animals↗
PubMed2004

Histological analysis and biological effects of granulation tissue around loosened hip prostheses in the development of osteolysis.

Although aseptic loosening of the prosthesis is a long-term complication after total joint replacement, the detailed mechanism of osteolysis remains unknown. We examined 82 samples from 40 patients with aseptic loosened hip prostheses histologically, and compared the distribution of particles, macrophages/histiocytes, and foreign body giant cells in the retrieved tissue from capsules and around prostheses. Furthermore, to investigate the mechanism of osteolysis, we cultured tissue from a patient with massive osteolysis and examined the effects of the conditioned medium on osteoblasts in vitro. Numerous multinucleated giant cells and histiocytes were present, and polyethylene particles ranging from medium to large were identified in the polarized light. However, the distribution was heterogeneous, and no particles were found microscopically in about 30%-40% of periprosthetic tissues, and in 60% of capsules. The amount of particles correlated with giant cells, but not with histiocytes. The conditioned medium of the granulation tissue culture stimulated osteoblasts to produce interleukin-6 in both protein and mRNA, and this was in part inhibited by anti-tumor necrosis factor-alpha or the interleukin-1beta antibody, suggesting that interleukin-6 production is mediated by several cytokines. These findings suggest that interleukin-6, which is produced not only by macrophages but also by osteoblasts, is a contributing factor to aseptic loosening.

Adult↗
PubMed2000

Detection of neurotropic viruses circulating in Tuscany: the incisive role of Toscana virus.

Acute meningitis is perhaps the most frequent among central nervous system infections. We report a study considering 277 cases of meningitis hospitalized in the southern Tuscany area (Italy) during the period from 1995 to 1998 investigated by tissue culture and polymerase chain reaction (PCR) methods. The cytochemical analysis of the cerebrospinal fluid samples suggested the diagnosis of aseptic meningitis, recognized as viral meningitis in 104 cases by detection of viral DNA or RNA. The results collected by tissue culture technique, available for 95 clinical samples, reported a positive isolation for only 12 cases. The viruses identified in the neurological infection were Toscana virus (81%), enterovirus (12%), mumps virus (3%), measles virus (1%), and herpes virus type 1 (3%). These data demonstrate the incisive role of the RNA viruses as the cause of meningitis, and overall the relevance of Toscana virus.

Adolescent↗
PubMed1994

Autologous cord blood transfusion.

Newborn piglets were exsanguinated (60% of blood volume) and retransfused 1 h later. One test group received adult pig red blood cells, the other piglet cord blood cells; controls were infused with plasma. While all controls died, satisfactory results were achieved in piglets transfused with either adult or foetal blood. The feasibility of collecting human cord blood for transfusion was assessed in 100 samples of human cord blood. Blood was collected aseptically and aerobic and anaerobic cultures set up. All samples of cord blood were sterile, and all were Mycoplasma negative. Coagulation parameters were analysed in eight cord plasma samples stored at -20 degrees C for 45 days. No significant abnormalities were found immediately after birth or after storage.

Animals↗
PubMed2004

Analysis of the serotype and genotype correlation of VP1 and the 5' noncoding region in an epidemiological survey of the human enterovirus B species.

The sequence identity of the enterovirus VP1 gene has been shown to correlate with the serotype concept. Enterovirus molecular typing methods are therefore often based on sequencing of the VP1 genomic region and monophyletic clustering of VP1 sequences of a homologous serotype. For epidemiological surveillance, 342 enterovirus samples obtained from patients with aseptic meningitis in Belgium from 1999 to 2002 were first diagnosed as being enterovirus positive by amplification of the 5' noncoding region (5'NCR) by reverse transcription (RT)-PCR. Subsequently, samples were molecularly typed by RT-nested PCR amplification and sequencing of a portion of the VP1 gene. Phylogenetic analyses were performed to investigate enteroviral evolution and to examine the serotype and genotype correlation of the two genomic regions. Our typing results demonstrated echovirus 30, echovirus 13, echovirus 18, and echovirus 6 to be the most predominant types. Echoviruses 13 and 18 were considered to be emerging human serotypes since 2000 and 2001, respectively, as they had been rarely reported before. Several serotypes existed as multiple genotypes (subtypes) from 1999 to 2002, but genomic differences mainly resided at synonymous sites; these results strongly suggest that the subtypes exhibit similar antigenic properties. Phylogenetic analyses confirmed that VP1 is an adequate region for molecular typing. Serotype-specific clusters are not observed commonly in phylogenetic trees based on the 5'NCR, and the phylogenetic signal in the 5'NCR was found to be particularly low. However, some substructure in the 5'NCR tree made a tentative prediction of the enterovirus type possible and was therefore helpful in PCR strategies for VP1 (e.g., primer choice), provided some background knowledge on the local spectrum of enteroviruses already exists.

Base Sequence↗
PubMed1998

Multicenter evaluation of the Amplicor Enterovirus PCR test with cerebrospinal fluid from patients with aseptic meningitis. The European Union Concerted Action on Viral Meningitis and Encephalitis.

The Amplicor Enterovirus PCR test was compared with viral culture for the detection of enteroviruses in cerebrospinal fluid (CSF) specimens. In a multicenter study in which nine laboratories participated, a total of 476 CSF specimens were collected from patients with suspected aseptic meningitis. Sixty-eight samples were positive by PCR (14.4%), whereas 49 samples were positive by culture (10.4%), demonstrating that the Amplicor Enterovirus PCR test was significantly more sensitive than culture (P < 0.001). After discrepancy analysis the sensitivity and specificity of the Amplicor Enterovirus PCR test obtained by using viral culture as the "gold standard" were 85.7 and 93.9%, respectively. Our results with the CSF specimens collected in different countries demonstrate that the Amplicor test is capable of detecting a large variety of enterovirus serotypes and epidemiologically unrelated isolates in CSF specimens from patients with aseptic meningitis. The Amplicor Enterovirus PCR test is a rapid assay which can be routinely performed with CSF samples and is an important improvement for the rapid diagnosis of enteroviral meningitis.

Cerebrospinal Fluid↗
PubMed1984

Serologic analysis of isolates of Pasteurella haemolytica and Staphylococcus aureus from mastitic ewes.

Milk samples (10 ml) were collected aseptically from infected and healthy mammary glands of 20 range ewes in the early stages of unilateral acute mastitis. The ewes were on 5 different ranches in the Northern Rocky Mountain region of the United States. The samples were plated on tryptose blood agar and examined for bacteria of possible etiologic significance. Twelve of the 20 ewes were infected with Pasteurella haemolytica, 4 ewes with Staphylococcus aureus, and 1 ewe with both bacteria. Twelve of the P haemolytica infections were in pure culture as were 4 S aureus infections. The 13 isolates of P haemolytica represented 6 different serotypes. Isolates of P haemolytica from ewes on the same ranch were as serologically diverse as were isolates from ewes in different herds. The 5 isolates of S aureus were similar antigenically. Bacterial isolates were not obtained from the milk of clinically healthy mammae.

Agglutination Tests↗
PubMed1998

High levels of expression of collagenase-3 (MMP-13) in pathological conditions associated with a foreign-body reaction.

A foreign-body-type host response can contribute to the induction and release of collagenolytic tissue-destructive enzymes of pathogenetic significance. Our aim was to analyse collagenase-3 in two conditions with putative involvement of foreign-body reactions. Synovial membrane-like tissue samples were obtained from cases of aseptic loosening of a total hip replacement (THR) and osteoarthritis (OA). The reverse transcription polymerase chain reaction (RT-PCR) disclosed that all the samples from patients contained collagenase-3 mRNA compared with only three out of ten control samples. The identity of the RT-PCR amplification product was confirmed by nucleotide sequencing. Immunohistochemical staining showed that collagenase-3 was present in endothelial cells, macrophages and fibroblasts, including those found in the synovial lining. This finding was confirmed by avidin-biotin-peroxidase complex-alkaline phosphatase-anti-alkaline phosphatase double staining and the specificity of the staining by antigen preabsorption using recombinant human collagenase-3. Collagenase-3 was released into the extracellular space and thus found in the synovial fluid in all patient samples as shown by Western blotting. The similar extent of collagenase-3 expression in aseptic loosening and OA compared with the low expression in control synovial membrane suggests involvement of a similar, foreign-body-based pathogenetic component in both. Comparative analysis of collagenase-3 and of foreign particles indicates that paracrine factors rather than phagocytosis per se are responsible for the induction of collagenase-3. We suggest that due to its localisation and substrate specificity, collagenase-3 may play a significant pathogenetic role in accelerating tissue destruction in OA and in aseptic loosening of a THR.

Adult↗
PubMed1999

Investigation of bacteremia after orthodontic banding.

The aim of this study was to assess the incidence of bacteremia after orthodontic banding. The study was conducted on 40 healthy orthodontic patients with good oral hygiene. Venous blood samples were obtained with a strict aseptic technique before and after fitting of a molar band in each patient. Microbiologic evaluation of the samples revealed a postoperative bacteremia incidence of 7.5%.

Adolescent↗
PubMed2004

Evaluation of IS900-PCR assay for detection of Mycobacterium avium subspecies Paratuberculosis infection in cattle using quarter milk and bulk tank milk samples.

A study was conducted to evaluate sensitivity, specificity, and predictive value of IS900-PCR assay for detection of Mycobacterium avium subspecies paratuberculosis in pooled quarter milk and bulk tank milk. Feces, blood and pooled quarter milk from 1493 lactating cattle on 29 herds were analyzed. Bulk tank milk (n = 29 bulk tanks) samples were also examined. Culture analysis revealed that 10.9%, 2.8%, and 20.6% of fecal, pooled quarter milk samples and bulk tanks were positive for M. avium subsp. paratuberculosis, respectively. While 13.5% and 27.5% of pooled quarter milk samples and bulk tanks were positive by IS900 PCR assay, respectively. Moderate to high antibody titers for M. avium subsp. paratuberculosis were detected in 223 of 1493 (14.4%) cows. Cows positive on fecal culture were taken as true positives relative to which the IS900 PCR assay was evaluated. The sensitivity and predictive value of KELA, pooled quarter milk culture, and IS900 PCR assay increased with lactation age. While the specificity of the tests decreased with increase in lactation age. Overall, the IS900 PCR assay using pooled quarter milk samples had a sensitivity, specificity and predictive value of 0.87, 0.95 and 0.71, respectively. The IS900 PCR assay using bulk tank milk had poor sensitivity (0.21), specificity (0.5) and predictive value (0.6). Pooled quarter milk culture analysis had a very low sensitivity (0.17). The kinetics ELISA had lower sensitivity (0.59), specificity (0.90) and predictive value (0.43) as compared to the IS900 PCR assay using pooled quarter milk samples. Results from our study suggest that IS900 PCR assay using bulk tank milk may not be useful for screening herds with M. avium subsp. paratuberculosis infected animals. In conclusion, use of IS900 PCR assay for cows in 2(nd) lactation and higher, using aseptically collected pooled quarter milk samples, can be a useful tool for screening and monitoring lactating cattle in herds with M. avium subsp. paratuberculosis infection.

Animals↗