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PubMed1981

[Effect of the sanitary state of the udder on the pyruvate content in the milk].

Studied was the pyruvate content of a total of 302 samples of "aseptic" milk as dependent on the clinical status of the mammary gland. The normal gland indices showed fluctuations from 0.48 up to 1.55 ppm at the respective cell element count, which ranged from 90000 to 480000 per cu. cm. It was found that when the secretory function of the gland was impaired the pyruvate level in the milk varied from 0.68 to 4.07 ppm at the respective cell count of 520000 to 4740000 per cm3. With milk from cows affected with subclinical mastitis the pyruvate value ranged from 0.87 to 6.13 ppm, and the cell count -- from 540,000 to 7,000,000. It is stated that the pyruvate test could not be used alone as a method in the evaluation of the clinical state of the udder, but could be referred to as an adjunct in the complex study of mammary gland.

Animals↗
PubMed1994

A cross-sectional sero epidemiological prevalence of polio virus antibodies post-immunization with three doses of TOPV.

A cross-sectional sero epidemiological study to detect the presence of antibodies to polio virus types 1, 2 and 3 was undertaken. A total of 437 infants with an average of 40 subjects per province was enrolled in this study. All the subjects had completed the three doses of TOPV. Blood samples were aseptically collected by heel pricking on calibrated filter papers and immediately transported to the laboratory for processing. In the laboratory, standardized techniques were used to detect neutralizing antibodies to polio virus. Antibodies with a titre of > 1:32 were detected in all studied subjects. Some infants developed titres as high as 1:1024. However, polio virus type 3 showed a higher antibody titre than the other two types. Differences in titres were observed from province to province.

Antibodies, Viral↗
PubMedDate not supplied

Efficacy of two avian Salmonella-immune lymphokines against liver invasion in chickens by Salmonella serovars with different O-group antigens.

Newly hatched chicks are susceptible to organ invasion by various serovars of Salmonella. We have previously shown that intraperitoneal administration of Salmonella enteritidis-immune lymphokine (SEILK) increased resistance to organ invasion by S. enteritidis (SE) in day-of-hatch chicks. In the present study, we determined that Salmonella-immune lymphokines can provide protection against organ invasion by serovars that are unrelated to the original serovar used as an immunogen. Immune lymphokines were produced from the splenic T cells of mature hens immunized with either viable SE or Salmonella anatum (SA). Day-of-hatch chicks were treated with either SA-immune lymphokine (SAILK) or SEILK and challenged per os with either Salmonella typhimurium (ST) or SE 1 hr later. Liver samples were aseptically collected 20 hr later and were cultured to detect salmonella. We found that SAILK conferred resistance against invasion by either SE, a serogroup D organism, or ST, a serogroup B organism. Furthermore, SEILK reduced organ invasion by ST. These observations together reveal that the protection against organ invasion conferred by Salmonella-immune lymphokines is unrelated to the serovar-specific antigenic characteristics of the immunizing bacteria.

Animals↗
PubMed1996

[Effects of different procedures of disinfection on the bacteriologic quality of hemodialysis monitors of the Cobe CS3 and AK 100 type].

]n recent years, hemodialysis treatment for chronic renal failure has been increasing. Although the technical evolutions have improved the therapy, the problem of microbial, toxic and chemical contamination of the dialysis fluid is now re-emerging. Most of all, because of increasing use of high-flux dialysis and its potential for transmembrane transport of bacteria into patients, one should be aware that dialysis fluid pathways may be colonised with bacteria. On the bacterial point of view, the French legislation proposed 100 CFU/mL as a maximum for total count of aerobic bacteria in the bi-osmosed water used for dilution of the dialysis concentrate. This study took place during 8 weeks in the children-hospital dialysis unit in Nancy (France). Our laboratory have succeeded in validating an aseptic bacteriological sampling procedure within fluid pathways of haemodialysis monitors Cobe CS3 and AK 100. It has also be shown that 6 to 12 hours of dialysis monitoring doesn't affect the quantitative and qualitative bacterial contamination of the biosmosed water (mean: 5 CFU/100 mL) drained through the 5 years' old internal pipes of the 2 monitors. So it has whatever the 4 different disinfection procedures applied and on the monitors (Formol or Formol + Citric acid + Chlorine or Heat or Heat + dehydrated Citric acid).

Child↗
PubMed1998

Detection of Leptospira DNA in patients with aseptic meningitis by PCR.

Samples of cerebrospinal fluid from 103 patients with aseptic meningitis were tested by PCR for detection of leptospires, and the results were compared with those of the microscopic agglutination test (MAT) and an enzyme-linked immunosorbent assay for detection of immunoglobulin M (ELISA-IgM). Of these samples, 39.80% were positive by PCR and 8.74 and 3.88% were positive by MAT and ELISA-IgM, respectively.

Adolescent↗
PubMed2001

Molecular detection and identification of an enterovirus during an outbreak of aseptic meningitis.

Stool samples from sixteen cases of children with meningitis originating from four different and geographically isolated parts of Greece were investigated for enteroviruses. The conventional method of cell culture in four different cell lines was initially used for the isolation of enteroviruses. The results showed a cytopathic effect (CPE) in all cases after two, or even more successive passages in only one cell line (RD), although a less-than-satisfactory CPE was obtained in many cases. Seroneutralization with RIVM mixed hyperimmune antisera followed and the isolates were typed as Coxsackie B viruses. The method of RT-PCR with enterovirus-specific primers targeted to the highly conserved 5'-UTR of the genome was initially used for the detection of enteroviruses from the inoculated cell cultures. A positive RT-PCR result was obtained for all of the clinical samples rapidly and accurately and the isolates were further characterized with the aid of Restriction Fragment Length Polymorphism (RFLP) analysis and Single Strand Conformation Polymorphism analysis (SSCP) of the amplicons. The RFLP analysis showed first of all that the isolates had an identical restriction pattern with Coxsackie B5 Faulkner reference strain with 4 out of 5 restriction enzymes and secondly, both RFLP and SSCP analysis indicated the epidemiological association of the isolates. The speed of the molecular methodology that was used in comparison with the conventional methods and its possible significance for the description of virus evolution and circulation in the populations is discussed.

Animals↗
PubMed2001

Effect of composition of colostrum and transition milk from Holstein heifers on specificity rates of antibiotic residue tests.

The objective of this study was to evaluate the effects of colostrum and transition milk composition on specificity rates of antibiotic residue screening tests. Milk from 25 primigravid Holstein heifers was collected from either first, second, or third milking (colostrum) and from either fifth, sixth, or seventh milking (transition milk) following parturition. Milk sampled was visibly normal and heifers were not treated with an antibiotic within 30 d before parturition. Quarter foremilk samples were collected aseptically and analyzed for mastitis pathogens. A sample from the total composite milk was analyzed for somatic cell counts (SCC), milk protein and fat, immunoglobulin concentrations and for antibiotics using four antibiotic residue screening tests. Mastitis pathogens were present in colostrum from 36% of heifers (n = 9) and from 16% of heifers (n = 4) in the subsequent transition milk. Mean SCC were 2,458,000 and 866,000 counts/ml and IgG1 concentrations were 22.7 and 3.07 mg/ml for colostrum and transition milk, respectively. Specificity rates of the screening tests ranged from 0.16 to 0.88 for colostrum and 0.60 to 1.0 for transition milk. Increased milk protein and IgG1 concentrations in milk were associated with an increase in the probability of a false positive outcome for the Charm Cowside (Charm Sciences, Inc., Malden, MA), CITE Snap (IDEXX Laboratories, Westbrook, ME), and Penzyme (Cultor Food Science, Milwaukee, WI) tests. Fat content of milk was positively related to an increase in false positive rates for the CITE Snap test. Milk should not be tested for antibiotic residues before the sixth milking after parturition to avoid high rates of false positive outcomes.

Animals↗
PubMed1989

Intrinsic and extrinsic factors influencing steroid production in vitro by bovine follicles.

Nonatretic bovine follicles were cultured on grids in a conventional static system, in roller tubes or in a continuous flux system. Culture medium from the static and roller tube system was replaced after 24 h of culture, while in the continuous flux system, only a small sample was taken aseptically at that time. Steroid concentrations in these samples were estimated by radioimmunoassay (RIA) and related to intrinsic factors like follicle size, day of cycle and micromorphological appearance at the end of culture and to extrinsic factors like the culture system, O(2)-concentration applied in the gasphase and time of culture. Bovine nonatretic follicles from 2 to 8 mm showed the same amount of estradiol-17beta (E2) production as pmol mg protein(-1) 24 h. Follicles over 8 mm had a significantly higher E2 production. Follicles from the follicular phase of the cycle produced more E2 than follicles from the luteal phase, independent of follicular size and culture system. Degeneration of follicles during culture resulted, independent of the culture system, in a decline of E2-production, and in an increase of P4-production; whereas the T-production initially (primary atresia) rose but subsequently (secondary and tertiary atresia) declined. The difference between the culture systems were reflected by quantitative differences in the production of the steroids measured. The most striking difference between the continuous flux system on one hand and the static and rolling tube system on the other is the predominant E2 production in the former by every follicle. It is thought that this difference might be caused by a better 02 supply in the continuous flux system. This hypothesis is tested in the static culture system. The more 02 the more E2 production. The increase in culture time resulted in an increase of E2 and P4, whereas the testosterone production was not significantly decreased.

Journal Article↗
PubMed1999

Investigations into the sterility of manually assembled extracorporeal circuits with vented reservoirs.

This study was designed to investigate the ability of an extracorporeal circuit (ECC) with a vented hard shell reservoir to remain sterile for a period of 72 h under dry conditions. The study was conducted in three phases. In Phase One: Two previously published methods for detecting contamination of the ECC were compared. A group of positive controls was collected by contaminating identical circuits with a known level of Enterobacter cloacae (ATTC: 13047) before initiating a regimen of "sample-dilute-sample" culturing. Negative controls for this phase were conducted by randomly sampling 1 L per manufacturer's lot of lactated ringers with each detection method. Culture results suggest that large volume filtration, but not small aliquot sampling, is sensitive to extremely low levels of contamination. No growth was detected in any negative control samples. In Phase Two: 19 ECC consisting of a membrane oxygenator, vented hardshell reservoir, arterial filter, and PVC tubing were removed from their sterile packages, assembled, and left unprotected in the moderate traffic environment of a research laboratory. The circuits were then primed with Lactated Ringer's solution. The prime solution was sampled for aerobic contamination by large volume filtration. None of the 19 samples detected contamination. In Phase Three: 43 ECC identical to the Phase Two circuits were assembled and left unprotected in the substerile pump room. The circuits were then primed, circulated, and cultured as in Phase Two. One of the 43 samples was discarded because of a recognized break in aseptic technique during sample collection. None of the remaining samples detected contamination. Mathematical calculations of binomial probabilities suggest that the chance of an open ECC developing a detectable level of contamination within 72 h of its dry assembly is insignificant.

Enterobacter cloacae↗
PubMed2006

Multiplex polymerase chain reaction detection of black-pigmented bacteria in infections of endodontic origin.

The purpose of this study was to detect the presence of Porphyromonas endodontalis, P. gingivalis, Prevotella intermedia, P. nigrescens, and P. tannerae from clinical samples using multiplex polymerase chain reactions (PCR). Two different multiplex PCR protocols were used (one for the two Porphyromonas species and the other for the three Prevotella species), each one using a primer pair specific for each target species. The results were compared to those of the conventional culture procedures. Microbial samples were taken aseptically from 40 infected root canals and abscesses from patients. Samples were cultured in an anaerobic condition for conventional identification using a Rapid ID 32 A kit. Multiplex PCR was processed using the DNA extracted from each sample. At least one of the five species of black-pigmented bacteria (BPB) were detected in 65% (26 of 40) of the samples using multiplex PCR, and in 15% (6 of 40) using the conventional culture procedures. Multiplex PCR was more rapid, sensitive, specific, and effective in detecting BPB than the conventional culture procedures.

Bacteroidaceae Infections↗
PubMed1993

The effect of glass-ionomer cement on carious dentine: an in vivo study.

The effect of a glass-ionomer cement sealant on carious dentine was investigated clinically and bacteriologically. The study group consisted of 24 molar teeth, with small clinical visible occlusal dentine lesions, in 13 children (aged 7-18 years). Twenty of these molars were filled and sealed with a glass-ionomer cement (Fuji Ionomer Type III) and 4 with a resin sealant (Delton). From each molar, two dentine samples were collected aseptically with a time interval of 7 months. The first sample was taken after opening the lesion just beneath the dentino-enamel junction before application of the filling material (sample A), and the second beneath the removed filling material (sample B) 7 months later. Before collecting sample B the sealant was clinically evaluated and impressions were prepared in order to (re)evaluate the sealants later by scanning electron microscopy (SEM). After taking the second sample, the remaining fissures were opened to a standard depth, the caries which was still present was removed, and a composite resin restoration was applied. In all teeth, more glass-ionomer cement material was found by SEM compared to the clinical evaluation. In both the glass-ionomer- and the resin sealant-treated group, the median value of the total number of microorganisms (CFU) on blood agar was 100 times smaller in the B sample. Microorganisms were found in 90% of the B samples in the group treated with glass-ionomer cement; hard dentine was also found in 45% of the B samples from this group.(ABSTRACT TRUNCATED AT 250 WORDS)

Actinomyces↗
PubMed1997

Effects of intravenous administration of sodium hyaluronate on carpal joints in exercising horses after arthroscopic surgery and osteochondral fragmentation.

OBJECTIVE: To evaluate the effects of arthroscopic surgery, osteochondral fragmentation, and treatment with IV administered hyaluronate on histologic, histochemical, and biochemical measurements within the carpal joints of horses. ANIMALS: 12 clinically normal horses, 2 to 7 years of age. PROCEDURE: Horses had an osteochondral fragment created at the distal aspect of the radiocarpal bone of 1 randomly chosen middle carpal joint to simulate osteochondral fragmentation. Horses were treated with 40 mg of hyaluronate or saline solution (placebo) intravenously once a week for 3 consecutive weeks (days 13, 20, and 27 after surgery). Treadmill exercise proceeded 5 days per week beginning 15 days, and ending 72 days, after surgery. Clinical evaluations were performed at the beginning and end of the study. Synovial fluid samples were obtained aseptically from both middle carpal joints on days 0, 13, 20, 27, 34, and 72 after surgery, and total protein, inflammatory cell, hyaluronate, glycosaminoglycan, and prostaglandin E2 concentrations were measured in each sample. All horses were euthanatized on day 72. Synovial membrane and articular cartilage were obtained for histologic evaluation. Articular cartilage samples were also obtained aseptically for determining glycosaminoglycan content and chondrocyte synthetic rate for glycosaminoglycans. RESULTS: Horses treated with hyaluronate intravenously had lower lameness scores (were less lame), significantly better synovial membrane histologic scores, and significantly lower concentrations of total protein and prostaglandin E2 within synovial fluid 72 days after surgery, compared with placebo-treated horses. Treatment with intravenously administered hyaluronate had no significant effects on glycosaminoglycan content, synthetic rate or morphologic scoring in articular cartilage, or other synovial fluid measurements. CONCLUSION: Intravenously administered hyaluronate appears to alleviate signs of lameness by interacting with synoviocytes, and by decreasing production and release of inflammatory mediators.

Animals↗
PubMed1985

Factors related to milk loss in quarters with low somatic cell counts.

Relationship between milk production and milk composition was studied through comparisons of udder halves within cow. Cows were milked by milking unit for separate quarters of udder. Six trials had six cows per trial. Trial length was 3 d, and milkings were at 12-h intervals. Foremilk samples were taken aseptically for bacterial analysis. Milk weights by quarter were recorded, and samples by quarter were analyzed for concentrations of lactose, somatic cells, and chloride. Milk cell differential counts and N-acetyl-B-D-glucosaminidase activity also were determined. Eighty-four percent of quarter milk samples contained less than 400,000 cells/ml. Differences between right and left udder halves with respect to all measurements were computed. For halves of udders within-cow correlation coefficients for differences between production and log(base 2) somatic cell count, lactose, chloride, bacterial presence, neutrophil percent, lymphocyte percent, macrophage percent, and N-acetyl-B-D-glucosaminidase activity were -.16, .23, -.31, .09, .12, .01, -.14, and -.41. Regression coefficients of milk production (kg) on somatic cell count log(base 2) cells per milliliter, lactose (%), chloride (mg/100 ml), and N-acetyl-B-D-glucosaminidase (nmol/min per ml) were -.12, .57, -.05, and -.46. From negative correlations between production and concentrations of chloride, somatic cells, and N-acetyl-B-D-glucosaminidase activity, differences between udder halves in production may be related to changes of the blood-milk barrier, leukocyte diapedesis, and loss of integrity of secretory cells.

Acetylglucosaminidase↗
PubMed2002

The effects of pentoxifylline on bacterial translocation after intestinal obstruction.

Bacterial translocation (BT) occurs mainly in preseptic conditions such as intestinal obstruction, trauma, and burn, and the underlying mechanisms are still unclear. Pentoxifylline (PTX) is a derivative of methyl xanthine and has several beneficial effects in sepsis. We investigated the effects of PTX on a rat BT model. Simple intestinal obstruction (IO) was choosen to create high BT rates. Rats were divided in to five groups of 10 rats. Either 50 mg/kg PTX or placebo (3 mg/100 g saline) was administered subcutaneously following IO, either by single injection or twice with a 12-h interval. All rats were sacrificed 12 or 24 h after the procedure, and mesenteric lymph nodes (MLN), liver, and blood samples were obtained under aseptic conditions for bacterial cultures. The samples were obtained 12 h following IO in the first two groups, and the same samples were obtained 24 h after IO in last three groups. Groups IV and V were the PTX treatment groups. PTX was re-injected 12 h after IO only in group IV. As a result, BT rates in MLNs and liver were found to be significantly low, blood specimens remained sterile in PTX-pretreated and -treated rats, and BT rates were high in control groups and group V (once treatment late specimen group). We conclude that simple intestinal obstruction causes BT, and PTX reduces BT in rats with IO during the first 12-h period if PTX is given once immediately following IO. PTX reduces BT during the first 24-h after IO provided that is injected twice with a 12-h interval. More experimental studies are need to explain the exact mechanism of this beneficial effect.

Analysis of Variance↗
PubMed1993

[Incidence of fungal isolation in clinical specimens from the respiratory tract].

The incidence of fungal isolation in 711 clinical specimens from the respiratory tract was investigated in 159 patients with pulmonary diseases. Fungi were isolated from 14/19 pharynx swab samples, 246/342 sputum samples, 34/147 bronchial aspirate samples, 18/97 bronchial lavage samples, 4/17 transbronchial lung biopsy samples, and 5/60 plugged catheter brush samples. Contamination by fungi in normal flora was prevented by using an aseptic procedure for sampling specimens from the respiratory tract. The isolation rate of Candida spp. from sputum was 70.0% (C. albicans: 151 strains, C. glabrata: 43 strains, C. tropicalis: 25 strains, C. lusitaniae: 4 strains, C. krusei: 3 strains, C. guilliermondii: 3 strains and C. paraspilosis: 1 strain). The isolation rates of C. neoformans and A. fumigatus from sputum were 6.4% and 8.8%, respectively. Isolated fungi from bronchial aspirate included 12 strains of C. albicans, 6 strains of C. tropicalis, 5 strains of A. fumigatus and 2 strains of C. neoformans. Isolated fungi from bronchial aspirate revealed 6 strains of C. albicans, 2 strains of C. glabrata, 1 strain of C. neoformans and 6 strains of A. fumigatus. The isolation rates of fungi from sputum obtained from patients with pulmonary aspergilloma and cryptococcosis were 66.5% and 50.0%, respectively. The isolation rate of fungi from sputum of patients with underlying diseases was higher than that of patients with no underlying disease.

Adolescent↗
PubMed2001

Presence and level of Campylobacter, coliforms, Escherichia coli, and total aerobic bacteria recovered from broiler parts with and without skin.

This study was undertaken to determine if broiler chicken parts without skin are less contaminated with Campylobacter than those with skin. Samples were taken in a commercial plant from defeathered carcasses before evisceration. Bacterial counts from rinse of aseptically removed meat samples were lower than those from stomached skin samples. No Campylobacter were recovered from meat collected from the breasts or thighs, and only 2 of 10 drumstick meat samples had detectable levels of Campylobacter. However, 9 of 10 breast skin, 10 of 10 thigh skin, and 8 of 10 drumstick skin samples were positive for Campylobacter, with between 2 and 3 log10 CFU/g of Campylobacter. Breasts, thighs, and drumsticks were removed from broiler carcasses following evisceration before entering the chill tank. There was a significant difference (50 to 90%) in the levels of Campylobacter on breasts, thighs, and drumsticks with and without skin. Similar trends were noted for coliform, Escherichia coli, and total aerobic bacterial counts from samples collected in the plant. Broiler part samples were also collected at retail outlets. These samples were either skin on and skinned in the laboratory or skin off at purchase. Aseptic removal of skin from broiler breasts, thighs, and drumsticks did not cause change in Campylobacter, coliform, E. coli, or total aerobic counts recovered from the skinned part. Likewise, parts purchased without skin did not have different bacterial counts than paired parts purchased with the skin on. Consumers should not expect to significantly lower the number of bacteria present on a chicken breast, thigh, or drumstick by removing the skin.

Animals↗
PubMed2001

Molecular typing of echovirus serotype 4 isolates.

We report on clinical samples Stuttgart/97, Berlin/99 and Jasi/99 associated with aseptic meningitis. All three samples contained echovirus 4 (E4) but Stuttgart/97 was simultaneous infected with echovirus 30 (E30). The genetic relationship of the E4 strains was assessed using RT-PCR and direct sequencing of amplicons derived from the genomic region encoding the capsid protein VP1. The sequences have been compared with each other and with sequences of further E4 strains obtained from GenBank. The analysis confirms that sequences of recent isolates have drifted away from elderly strains over a longer period of time. Several amino acid changes in assumed antigenic sites of the VP1 gene may be sufficient to cause changes in antigenic specificity and therefore they may be a reason for failure of serological typing of some new antigenic E4 variants.

Amino Acid Sequence↗
PubMed1983

Facultative gram-negative enteric rods in persistent periapical infections.

A bacteriological investigation has been made in two cases of persistent periapical infections. Neither of the two infections responded to root canal therapy including penicillin or penicillin and erythromycin. Samples were taken aseptically for bacteriological examination through and root canals. Chloroform-washed, sterile paper points were used for sampling. Cultivation was performed immediately at chairside on Kanamycin-Vancomycin laked blood agar (KVLB) and blood agar supplemented with menadione, cysteine and glucose for anaerobic incubation and chocolate agar for aerobic incubation. Enterobacter cloacae was only isolate in case 1; Klebsiella pneumoniae and enterococci were found in case 2. Obligately anaerobic bacteria were not found. The treatment was successful after antibiotic therapy based on susceptibility testing of the isolates. The route of infection by facultative enteric rods is discussed.

Adult↗