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PubMed2002

Evaluation of bacteriologic culture of pooled fecal samples for detection of Mycobacterium paratuberculosis.

OBJECTIVES: To compare sensitivity of several methods of bacteriologic culture of pooled bovine fecal samples for detection of Mycobacterium paratuberculosis and evaluate homogeneity in number of M paratuberculosis in pooled fecal samples. SAMPLE POPULATION: Feces from 10 dairy cows that shed M paratuberculosis at various concentrations and 1 dairy cow known to be free of infection with M paratuberculosis. PROCEDURE: 5 fecal pooling methods, 2 culture methods, and 2 pool sizes were evaluated. Each pooled sample contained 1 infected sample and 4 or 9 uninfected samples. RESULTS: Sensitivity of detection of M paratuberculosis was greater with smaller pool size (5 vs 10 samples/pool). Detection sensitivity was also associated with concentration of bacteria in the infected sample. Results indicated that, compared with concurrent bacterial culture of individual infected samples, 37 to 44% of pooled samples with low bacterial concentrations yielded positive culture results and 94% of pooled samples with high bacterial concentrations yielded positive results. CONCLUSIONS AND CLINICAL RELEVANCE: Bacteriologic culture of pooled fecal samples may provide a valid and cost-effective method of detecting M paratuberculosis infection in cattle herds.

Animals↗
PubMed2005

False-positive outcome and drug residue in milk samples over withdrawal times.

This study was conducted to identify false-positive outcomes and drug residues in milk samples over withdrawal times and to determine whether the positive results were caused by drug residues or natural inhibitors. A total of 73 milk samples over withdrawal times after the last intramammary infusion were collected from each treated quarter of cows and tested using the Delvotest SP assay. Reading time was 150, 165, and 180 min, and results of samples were recorded according to the color of the well containing the control milk sample. There were 24, 20, and 12 positive samples at the reading times of 150, 165, and 180 min, respectively. All 24 positive milk samples were heated at 82 degrees C for 5 min and retested to verify that the positive results were caused by drug residues or natural inhibitors. Twenty-one samples that exhibited positive results were negative after heat treatment, and drug residues were not identified by LacTek and Charm tests. However, 3 samples that exhibited positive results from heat treatment of 82 degrees C were positive for drugs. In our study, most positive results (89%) in the milk samples over withdrawal times were false-positive results by natural inhibitors. Moreover, the heat treatment is a fast, simple, and inexpensive method to remove false-positive results and has no effect on positive samples containing drugs. We suggest that heat treatment before screening tests is an effective way to reduce false-positive results in the milk samples.

Animals↗
PubMed2006

Evaluation of environmental sampling and culture to determine Mycobacterium avium subspecies paratuberculosis distribution and herd infection status on US dairy operations.

The objectives of this study were to determine the distribution of Mycobacterium avium subspecies paratuberculosis (MAP) in the environment and assess the relationship between the culture status of MAP in the farm environment and herd infection status. The National Animal Health Monitoring System's Dairy 2002 study surveyed dairy operations in 21 states. One component of the study involved collection and culturing of environmental samples for MAP from areas on farms where manure accumulated from a majority of a herd's cows. Operations were selected for inclusion based on perceived risk factors for MAP infection identified in a previously administered questionnaire. Individual animal and environmental samples were collected and used to determine the efficiency of environmental sampling for determination of herd infection status. Individual animal fecal, serum, and milk samples were used to classify herds as infected or not infected based on the presence of at least one test-positive animal in the herd. A total of 483 environmental samples (approximately 5 per farm) were collected, and 218 (45.1%) were culture-positive for MAP. A similar percentage of environmental cultures collected from all designated areas were positive [parlor exits (52.3%), floors of holding pens (49.1%), common alleyways (48.8%), lagoons (47.4%), manure spreaders (42.3%), and manure pits (41.5%)]. Of the 98 operations tested with the environmental sample culture, 97 had individual serum ELISA results, 60 had individual fecal culture results, and 34 had individual milk ELISA results. Sixty-nine of the 98 operations (70.4%) had at least one environmental sample that was culture-positive. Of the 50 herds classified as infected by fecal culture, 38 (76.0%) were identified by environmental culture. Two of the 10 operations classified as not infected based on individual animal fecal culture were environmental culture-positive. Of the 80 operations classified as infected based on serum ELISA-positive results, 61 (76.3%) were identified as environmental-positive, whereas 20 of the 28 (71.4%) operations identified as infected based on milk ELISA were detected by environmental sampling. Environmental sample culturing is less costly than individual animal sampling, does not require animal restraint, and identified more than 70% of infected operations. Environmental sampling is another diagnostic tool that veterinarians and dairy producers can use to determine herd infection status for MAP.

Animals↗
PubMed1991

Near infrared spectroscopic analysis of forage samples digested in situ (nylon bag).

The objective was to investigate the feasibility of using near infrared reflectance spectroscopy to determine the composition of samples generated in situ. Five alfalfa and five orchardgrass hays of differing maturities were incubated for 0, 6, 24, 48, and 72 h (Experiment 1) or for 0, 2, 4, 8, 12, 24, 48, and 96 h (Experiment 2) in rumen-fistulated, lactating cows, using nylon bags. After washing to remove rumen contents, samples were analyzed using a nonrotating circular cell in a scanning monochromator. All samples then were dried at 55 degrees C and analyzed for CP and ADF by wet chemistry and rescanned in the dry state. The degree of DM digestion of the original sample was calculated from duplicate bags. Results for spectral analysis of dried samples (Experiment 1), with one-half the samples for validation, were typical of results found for dry forages. The results for scanning wet samples were less accurate than for dry ones. Analysis of samples from Experiment 2 by equations developed in Experiment 1 often resulted in extremely large biases, but these were corrected by including six samples of each forage from Experiment 2 in the calibration set (from Experiment 1) and redeveloping the equations. Although it is possible to use near infrared reflectance spectroscopy to determine the composition of wet samples generated in situ, results are more accurate if the samples are scanned after drying.

Animal Feed↗
PubMed1986

Determination of crude protein and fat in carcass and breast muscle samples of poultry by near infrared reflectance spectroscopy.

Near infrared reflectance spectroscopy (NIR) equipment used consisted of an InfraAlyzer 400+ fitted with 19 discrete filters and an HP-85 microcomputer. Calibrations were developed with chicken broiler carcass samples. Samples were divided into two sets: a calibration (CAL) from which equations were developed and a prediction (PRE) set used to validate the equations. The CAL and PRE sets in the carcass calibration consisted of 45 and 54 samples, respectively. Two equations were developed for the prediction of fat in carcass samples (FAT1 and FAT2). Calibration for breast muscle samples included 30 and 27 samples for CAL and PRE sets, respectively. The accuracy of the calibrations was measured by the coefficient of determination (r2), standard error of the estimate (SEE), coefficient of variation (CV), and the mean difference between NIR and chemical values (d). Repeatabilities of chemical and NIR procedures were determined by the F-ratio of the variance of the differences between replicates. The result of the regression analysis between chemical and NIR values indicated good predictions for crude protein (CP) and fat in carcass samples. The r2 and SEE for CP, FAT1, and FAT2 were .98, .94; .91, 2.48; and .91, 2.51 in CAL set; and .91, 1.03; .68, 2.32; and .69, 2.30 in PRE set samples, respectively. No differences were found between FAT1 and FAT2 equations. The carcass calibration equations were tested with samples from laying hens. Higher SEE were found for CP (1.78), but SEE for fat were similar to broiler carcasses. The calibration for breast samples showed higher SEE for CP and fat when compared with carcass samples.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Sampling methods in behavior research.

Animals perform a continuous stream of behavior throughout their lives. Because their behavior is not random, appropriate sampling methods can be used to obtain data that accurately reflect the actual behavior and are valid for answering research questions. Answering questions related to several variables assists in narrowing the choices of sampling methods. First, a determination must be made of what behaviors to measure. If the behaviors are few and easily measured, then All Occurrences Sampling is the method of choice because it generates accurate frequency and duration data through continuous recording. Sequence and Sociometric Matrix Sampling are specialized types of All Occurrences Sampling that are restricted to sampling intra- or interindividual sequences and social interactions (e.g., agonistic), respectively. Second, if who (e.g., specific individual, sex, or genotype) performs the behavior is a major component of the research question, then consideration should be given to Focal Animal (Pair, Group) Sampling. Third, if when or where the behavior is performed is of interest (e.g., activity budget), then Instantaneous or Scan Sampling can often be effective. Ad libitum Sampling does not produce valid data for analyses, but it is useful when formulating and fine-tuning research questions. One-Zero Sampling is not recommended except when the research question relates to the presence or absence of behaviors only. Other factors to consider in selecting a sampling method are duration of the behavior (event or state), desired scale of measurement (nominal, ordinal, interval, or ratio), and logistics (e.g., time, and equipment and facilities available).

Animals↗
PubMed2001

Difficulty in recovering inoculated Campylobacter jejuni from dry poultry-associated samples.

We inoculated 5 cm2 of clean chick pads, 5 g of clean pine shavings, and fresh unsanitized broiler breeder eggshell halves with a cell suspension of Campylobacter jejuni in physiological saline. Inoculation levels were 10(2), 10(3), or 10(4) cells per sample. The samples were allowed to remain at room temperature for 15, 30, or 60 min before addition of enrichment broth. When chick pad samples were inoculated with 102 cells, by 15 min 40% of the samples had detectable levels of Campylobacter, and by 30 to 60 min Campylobacter could be detected in only 20% of the samples. With samples of pine shavings, only 25% of those inoculated with 103 cells were positive for Campylobacter after 15 min and only 5% were positive for Campylobacter after 30 min. When 104 cells were inoculated onto litter, Campylobacter was recovered from 20% of the samples at 15 min and 15% of the samples after 30 min. Eggshells were also found to be a harsh environment. When the inoculum was 102 at 15 min, 8 of 10 samples were positive for Campylobacter but at 60 min only 10% of the samples remained positive for Campylobacter. The current cultural methods may not be adequate for recovering low numbers of Campylobacter from dry samples. Campylobacter may be present but culturally undetectable in the commercial hatchery and hatchery environment.

Animals↗
PubMed1998

Clinical relevance of in vitro propagation of activated lymphocytes from endomyocardial biopsy samples of pediatric heart transplant recipients.

In vivo activated T-lymphocytes can be cultured from endomyocardial biopsy samples of human cardiac allografts, sometimes even in the absence of histological rejection. We investigated the clinical relevance of this "lymphocyte growth assay" in pediatric heart transplant recipients. Specifically, we wished to determine if: (i) positive lymphocyte growth from EMB samples in the absence of significant rejection identifies a patient as being at increased risk for the development of acute rejection; (ii) withdrawal or major dose reduction of corticosteroids in the presence of lymphocyte growth results in high risk of rebound rejection; and (iii) presence of lymphocyte growth during acute rejection helps predict the response to treatment. Cultures were performed on 789 consecutive EMB samples from 65 pediatric heart transplant recipients in media containing 30 U/ml of recombinant IL-2. T-lymphocytes were cultured from 16% of EMB samples with low grade rejection (grade 0-1b) and from 34% of EMB samples with grade 2-4 rejection. EMB samples obtained early post-transplant (<180 days) were significantly more likely to yield positive lymphocyte growth compared to biopsies obtained late for any given rejection grade. Lymphocyte growth was comparable between patients managed with cyclosporine or tacrolimus based immunosuppression. For 227 EMB samples without rejection, a subsequent EMB sample was obtained within 12 weeks. Lymphocyte cultures were positive in 47 of these 227 EMB samples (21%), and in 19 out of 47 (40%) cases acute rejection (grade 2-4) was present on the follow-up EMB sample. By contrast, of 180 biopsies without growth, only 29 (16%) showed rejection at the next EMB (p<0.0001). When a follow-up biopsy was performed within 12 weeks of corticosteroid withdrawal, "rebound rejection" was observed in 3 out of 10 (30%) cases where the previous EMB sample yielded positive lymphocyte growth and in 4 out of 38 (11%) cases when it did not (p=0.29). The presence of lymphocyte growth in association with rejection was also predictive of whether rejection would resolve following high dose intravenous corticosteroid therapy (persistent rejection in 33 out of 50 (66%) cases with positive growth, versus 25 out of 80 (31%) cases without growth (p<0.0001)). Thus, positive lymphocyte growth is strongly associated with higher grade of rejection and earlier time from transplantation. Lymphocyte growth in the absence of rejection indicates high risk for rejection within the next 12 weeks. Growth in association with acute rejection indicates high probability of persistence of rejection following treatment with high dose corticosteroids.

Adolescent↗
PubMed1999

Analytical characterization of electrochemical biosensor test strips for measurement of glucose in low-volume interstitial fluid samples.

BACKGROUND: Minimally invasive interstitial fluid (ISF) sampling and glucose measurement technologies were integrated into a hand-held device for diabetic glucose monitoring investigations. METHODS: Conventional electrochemical test strip technology (Bayer Glucometer Elite) was adapted to measure glucose in small (0.5-2.0 microL) samples of ISF. Test strip glucose measurements were performed on a commercial potentiostat and were compared to various reference glucose methodologies (YSI 2300 analyzer, microhexokinase procedure, Bayer Glucometer Elite). Characterizations of the integrated ISF sampling-glucose test strip design included accuracy and precision in various sample media (saline, ISF surrogates, diabetic ISF samples), sample volume dependence, test strip sterilization studies (electron beam, gamma irradiation), and diabetic ISF sampling and glucose measurements. RESULTS: Glucose measurements were free from significant media effects. Sample volume variations (0.6-3.2 microL) revealed only modest dependence of glucose measurement bias on sample volume (-1.5% per microliter). Sterilization treatments had only a minor impact on glucose response and test strip aging and no significant impact on interferent responses of the glucose test strips. Diabetic subject testing under minimum fasting conditions of at least 2 h with integrated ISF sampling and glucose measurement gave low ISF glucose measurement imprecision (CV, 4%) and mean glucose results that were indistinguishable from reference (microhexokinase) ISF glucose measurements and from capillary blood glucose measurements (Glucometer Elite). CONCLUSIONS: Conventional single-use, electrochemical glucose test strip and ISF collection technologies can be readily integrated to provide real-time ISF sampling and glucose measurements for diabetic monitoring applications.

Biosensing Techniques↗
PubMed2000

Simplification for measuring input function of FDG PET: investigation of 1-point blood sampling method.

UNLABELLED: The current method for quantitative FDG PET study requires application of multiple arterial blood sampling for measuring the input function, but the procedure is invasive and complicated. The purpose of this study was to establish a 1-point blood sampling technique that gives data comparable with the data of more elaborate serial arterial sampling. METHODS: We established a time point for 1-point arterial sampling that exhibited the highest correlation between plasma radioactivity at the time point and the real integrated value (IV) of the measured input function obtained by multiple arterial sampling in 120 patients and the smallest coefficient of variation of the real IV divided by plasma radioactivity at the time point in 120 patients. Scaling factors for estimation at each sampling point were determined, and a reference table was established to make the supposed input function. RESULTS: The optimal time for 1-point arterial sampling was 12 min after FDG injection. A good correlation was observed between the real IVs and those estimated from 1-point arterial blood sampling at 12 min using the supposed input function (n = 120; P < 0.001). The time point at which the difference between values of arterial and venous blood disappeared was 40 min after FDG injection. The percentage errors of IV estimation by 1-point sampling were 1.70% (n = 120) for arterial blood at 12 min and 3.64% (n = 10) for venous blood at 40 min. CONCLUSION: We conclude that the simplified 1-point sample method works in a manner that is comparable with serial arterial sampling and should be useful for clinical PET.

Adolescent↗
PubMed1975

A quantitative sampling method for Oncomelania quadrasi by filter paper.

Filter paper was found to attract Oncomelania quadrasi in waters the same way as fallen dried banana leaves, although less number of other species of snails was collected on the former than on the latter. Snails were collected in limited areas using a tube (85 cm2 area at cross-section) and a filter paper (20 X 20 CM) samplers. The sheet of filter paper was placed close to the spot where a tube sample was taken, and recovered after 24 hours. At each sampling, 30 samples were taken by each method in an area and sampling was made four times. The correlation of the number of snails collected by the tube and that by filter paper was studied. The ratio of the snail counts by the tube sampler to those by the filter paper was 1.18. A loose correlation was observed between snail counts of both methods as shown by the correlation coefficient r = 0.6502. The formulas for the regression line were Y = 0.77 X + 1.6 and X = 0.55 Y + 1.35 for 3 experiments where Y is the number of snails collected by tube sampling and X is the number of snails collected in the sheet of filter paper. The type of snail distribution was studied in the 30 samples taken by each method and this was observed to be nearly the same in both sampling methods. All sampling data were found to fit the negative binomial distribution with the values of the constant k varying very much from 0.5775 to 5.9186 in (q -- p)-k. In each experiment, the constant k was always larger in tube sampling than in filter paper sampling. This indicates that the uneven distribution of snails on the soil surface becomes more conspicuous by the filter paper sampling.

Animals↗
PubMed2002

Linkage for platelet monoamine oxidase (MAO) activity: results from a replication sample.

BACKGROUND: Monoamine oxidase B (MAO-B) degrades catecholamines in presynaptic nerve endings and is also active in platelets. There is evidence to suggest that platelet MAO-B activity level is controlled by a major genetic locus distinct from the structural gene on the X chromosome. To expand on a prior report, new linkage analyses for platelet MAO-B activity have been performed on the previously analyzed sample (designated the initial sample), on a new sample of families (the replication sample), and on the combined sample. These families were recruited as part of the Collaborative Study on the Genetics of Alcoholism (COGA). METHODS: The initial sample consists of 105 extended families providing 1002 nonindependent (412 independent) sib pairs that have been phenotyped for MAO activity and genotyped. The replication sample of 157 extended families contains 608 nonindependent (309 independent) phenotyped and genotyped sib pairs. Analyses were conducted using Haseman-Elston based regression on sib pairs and variance component analysis on extended pedigrees, and the importance of cigarette smoking and gender as covariates of platelet MAO-B activity was taken into account. RESULTS: Regions on chromosomes 2, 9, and 12 indicated consistent evidence for linkage across the two distinct datasets by at least one analysis method. Under Haseman-Elston regression of independent sib pairs, only the chromosome 2 region gave lod scores above 1 in both the initial and replication samples. Using all possible pairs, unweighted, for the regression, chromosome 12 gave lod scores above 1 in both samples. For variance component analysis, only the chromosome 9 region gave lod scores above 1 in both samples. CONCLUSIONS: The consistency across datasets of these findings is encouraging. In particular, variance component analysis of extended pedigrees supports a potential linkage of MAO-B activity to chromosome 9, with a lod over 3 at 115 cM near D9S261 in the combined sample. Sib-pair regression supports this finding with modest lod scores in the region. Suggestive linkage to chromosomes 2 and 12 from sib-pair analysis is only weakly supported by variance component analysis.

Blood Platelets↗
PubMed2002

Stabilization of mRNA expression in whole blood samples.

BACKGROUND: Accurate quantification of mRNA in whole blood is made difficult by the simultaneous degradation of gene transcripts and unintended gene induction caused by sample handling or uncontrolled activation of coagulation. This study was designed to compare a new blood collection tube (PAXgene Blood RNA System) and a companion sample preparation reagent set with a traditional sample collection and preparation method for the purpose of gene expression analysis. METHODS: We collected parallel blood samples from healthy donors into the new sample collection tubes and control EDTA tubes and performed serial RNA extractions on samples stored for 5 days at room temperature and for up to 90 days at 4 and 20 degrees C. Samples were analyzed by Northern blot analysis or reverse transcription-PCR (RT-PCR). RESULTS: Specific mRNA concentrations in blood stored in EDTA tubes at any temperature changed substantially, as determined by high-precision RT-PCR. These changes were eliminated or markedly reduced when whole blood was stored in PAXgene tubes. Loss of specific mRNAs, as measured by RT-PCR, reflected total RNA depletion as well as specific mRNA destruction demonstrated by Northern blot analysis. The salutary effects of PAXgene on mRNA stabilization extended to blood samples from eight unrelated donors. CONCLUSIONS: Compared with whole blood collected in EDTA tubes and extracted by an organic method, the PAXgene Blood RNA System reduced RNA degradation and inhibited or eliminated gene induction in phlebotomy whole blood samples. Storage of whole blood samples in PAXgene tubes can be recommended for clinically related blood samples that will be analyzed for total or specific RNA content.

Blood Specimen Collection↗
PubMed2004

[Comparison of three methods for quantitative analysis of LPO in different biological samples].

OBJECTIVE: To explore the optimized methods for detecting lipid peroxide (LPO) in biological samples and the reference value of LPO in human plasma. METHODS: Three most commonly adopted methods were used for detecting LPO in different biological samples simultaneously, and then their linearity, accuracy, precision, stability and detecting efficiency were compared. The methods were FOX assay, Modified iodometric assay and TBARS assay. The standard curve (linearity evaluation), rate of sample recovery (accuracy evaluation), reproducibility (precision evaluation), stability of reading number (stability evaluation), as well as the detected values of LPO in different sample systems by three methods simultaneously (detecting efficiency) were evaluated. The sample systems were: isolated low-density lipoprotein (LDL), supernatant of cell culture, and human plasma. RESULTS: When applied to detecting LPO in the isolated LDL sample system, FOX assay was found to have the most sensitive detecting efficiency, good accuracy and precision. When applied to detecting LPO in the supernatant of cell culture and human plasma sample systems, the Modified iodometric assay and TBARS assay showed better function than FOX assay; the rate of sample recovery of FOX assay 61.92% +/- 2.92% was obviously lower as compared with 99.00% +/- 2.65% of modified iodometric assay and 101.63% +/- 12.00% of TBARS assay; and the reproducibility of FOX assay 19.15% was also lower as compared with 4.36% of Modified iodometric assay and 3.14% of TBARS assay. The three methods all showed fine linearity and stability. The values of LPO concentration in normal human plasma were (14.189 +/- 4.889) mumol/L by Modified iodometric assay and (0.936 +/- 0.462) mumol/L by TBARS assay; these values were close to those in other reports. CONCLUSION: FOX assay was found to be most sensitive in the three methods for measurement of LPO in a relative pure sample system (such as isolated LDL). In complex sample system, however, the Modified iodometric assay and TBARS assay showed better function. The authors suggest that suitable method be chosen according to the nature of sample, that more than one method be chosen for plasma LPO assay in the same planned analysis, and that Modified iodometric assay and TBARS assay be worth the first choice.

Clinical Chemistry Tests↗
PubMed2005

[PCR detection of PCV-2 and PRRSV in porcine pleuropneumonia samples].

PCV-2 (Porcine circovirus type-2, PCV-2) and PRRSV (Porcine reproductive and respiratory syndrome virus, PRRSV) were detected by PCR from 253 porcine pleuropneumonia samples and 125 clinically healthy lung samples were collected from different districts of Shandong province. The results showed that 171 samples for PCV-2 and 101 samples for PRRSV were positive. The positive ratio were 67.5% and 40%, respectively. The co-infection number of PCV-2 and PRRSV was 68 in 253 samples, the positive ratio was 26.8%. While in the clinically healthy samples, only 21 samples for PCV-2 and 12 samples for PRRSV were detected positive, the positive ratio were 16.8% and 9.6%, respectively, no co-infection samples were found. Statistical result showed significant difference between positive ratio for PCV-2 and PRRSV in porcine pleuropneumonia samples and that of in clinically healthy samples. The above results demonstrated that there maybe some relationship between the infection of porcine pleuropneumonia and PCV-2 and PRRSV in pigs.

Animals↗
PubMed2005

Evaluation of Western blotting methods using samples with or without sodium phosphotungstic acid precipitation for diagnosis of scrapie and chronic wasting disease.

The purpose of this study was to enhance the sensitivity of the Western blot (WB) test for use as an alternative and confirmatory method for the diagnosis of scrapie and chronic wasting disease (CWD) in Canada by comparing 2 sample preparation procedures: an abnormal prion protein (PrPSc) concentration procedure using sodium phosphotungstic acid (PTA) precipitation and a procedure using crude sample without precipitation. A total of 100 cerebrum samples (52 sheep and 48 elk), including 66 negative (31 sheep, 35 elk) and 34 positive (21 scrapie and 13 CWD positive) samples diagnosed by using immunohistochemistry (IHC) on retropharyngeal lymph node (RPLN) and medulla oblongata at obex, were tested by using WB with the 2 sample preparation procedures. The WB using non-PTA enriched sample (crude extract) detected, on average, only 71.7% (9 of 15, 60.0% for scrapie, 5 of 6, 83.3% for CWD) of the samples that tested positive by using WB with PTA enriched samples. No case was positive by WB using crude extract but negative by WB using PTA enriched sample. No false positive was found. Serial dilution of PTA precipitated samples demonstrated that the technique increases the detection limit approximately 100 fold. Additionally, the comparison of the WB and IHC on cerebrum from all the positive cases demonstrated that WB following PTA precipitation and IHC had 100% agreement by detecting 6 positive for CWD on cerebrum; while IHC detected scrapie in only 14 out of 15 positive cerebrum samples by using WB following PTA precipitation. Phosphotungstic acid precipitation is therefore a useful adjunct to WB analysis of scrapie and CWD and tissues.

Animals↗
PubMed2004

Prevalence of Aeromonas in different clinical and water samples with special reference to gastroenteritis.

A hospital based study was carried out in the Department of Microbiology, Tribhuvan University Teaching Hospital, Institute of Medicine with the aim to initiate the isolation and identification of Aeromonas spp. from the stool samples of gastroenteritic patients and to determine the prevalence of Aeromonas spp. in other clinical samples and water. Altogether 293 samples were investigated that include 172 stool samples, 60 pus/wound swabs, 20 body fluids and 41 water samples. The samples were collected and processed by standard microbiological techniques in order to isolate Aeromonas. Ampicillin blood agar (20 microg/ml) was used as selective medium for the isolation of Aeromonas. The specimen prevalence rate of Aeromonas spp. in stool was found to be 5.2% and the A. hydrophila (55.5%) was the predominant species followed by A. caviae (33.3%) and A. sobria (11.1%) in the stool samples. Likewise, 3.3% of pus sample showed positive growth of A. hydrophila. Aeromonas was not detected in any of the body fluids. Aeromonas spp. were isolated from 58.5% of water samples obtained from different hospitals. The commonest species was A. hydrophila (62.5%) followed by A. caviae (20.8%) and A. sobria (16.7%). In vitro susceptibility testing showed that the aminoglycosides and fluroquinolones were the effective antibiotics against Aeromonas. It was found that 88.9% of Aeromonas spp. isolated from stool samples were sensitive to gentamicin, 77.8% to ciprofloxacin, norfloxacin, tetracycline and ceftriaxone and 66.6% to nalidixic acid whereas cent percent Aeromonas spp. from water samples were sensitive to gentamicin, ciprofloxacin, norfloxacin, tetracycline and ceftriaxone and 75.0% to nalidixic acid. The enteric Aeromonas isolates were more resistant to antimicrobial agents than the water isolates.

Adolescent↗
PubMed1992

[Testing Milkofix, a new preservative preparation for milk samples used for infrared analysis of milk components. II. Verification of its preservative effects in relation to infrared analysis].

For the purposes of the infrared analysis of the basic milk composition, Milkofix, an ecologically friendly preparation used for milk sample preservation (Trzický, 1990), was compared with untreated samples (N) and with samples preserved with sodium azide (A), bronopol (B) and potassium dichromate (C) at a storage temperature of 20 degrees C (I) and 4 degrees C (II) in samples kept for 14 and 18 days. Pursuant to the recommendations cited in literature, the preservatives had these concentrations: A = 0.0085 g NaN3 and 0.0630 g NaCl; B = 0.0050 g bronopol and 0.0500 g NaCl; C = 0.0330 g K2Cr2O7 and 0.0670 g KCl; M = 0.1250 g, all amounts are per 25 ml milk. Three bulk milk samples were used which were analyzed on an automatic Milko-Scan 133 B infraanalyzer (Foss Electric, Denmark) every day. On the basis of a graphical evaluation of the results by IDF recommendations (1985) the times within which the applicable results could be obtained were determined for the various methods of milk sample treatment (Figs. 1 to 6): N I--0 days; A I--9; B I--10; C I--13; M I--4; N II--10; A II--5; B II--11; C II--15; M II--10 (Tab. I). The results recorded for Milkofix are in agreement with the conclusions drawn in the previous study, where the intervals of four and nine days were determined. The days to milk sample coagulation were as follows: N I--1 day, M I--10 days. The coagulation in A I, B I, C I and N II samples was not observed even after 13-day storage and in A II, B II, C II and M II samples not even after 17 days of storage. The results for particular components (fat, proteins, lactose) of milk samples differently treated in time are presented in Tabs. II, III and IV. A system of evaluating criteria (Tab. V) was used to determine the order beginning from the most convenient method of milk sample treatment for the given purpose: 1. C II, 2. C I, 3. B II and A I, 4. B I, 5. M II, 6. N II, 7. A II, 8. M I and 9. N I.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗