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Sample records for cattle serum samples

  1. A Novel Genetic Group of Bovine Hepacivirus in Archival Serum Samples from Brazilian Cattle

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    Cláudio W. Canal

    2017-01-01

    Full Text Available Hepatitis C virus (HCV (genus Hepacivirus; family Flaviviridae is a major human pathogen causing persistent infection and hepatic injury. Recently, emerging HCV-like viruses were described infecting wild animals, such as bats and rodents, and domestic animals, including dogs, horses, and cattle. Using degenerate primers for detecting bovine pestiviruses in a 1996 survey three bovine serum samples showed a low identity with the genus Pestivirus of the Flaviviridae family. A virus could not be isolated in cell culture. The description of bovine hepaciviruses (BovHepV in 2015 allowed us to retrospectively identify the sequences as BovHepV, with a 88.9% nucleotide identity. In a reconstructed phylogenetic tree, the Brazilian BovHepV samples grouped within the bovine HCV-like cluster in a separated terminal node that was more closely related to the putative bovine Hepacivirus common ancestor than to bovine hepaciviruses detected in Europe and Africa.

  2. Evaluation of an indirect ELISA for the diagnosis of bovine brucellosis in milk and serum samples in dairy cattle in Argentina.

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    Vanzini, V R; Aguirre, N; Lugaresi, C I; de Echaide, S T; de Canavesio, V G; Guglielmone, A A; Marchesino, M D; Nielsen, K

    1998-09-01

    An indirect enzyme-linked immunosorbent assay (ELISA) for Brucella abortus antibodies detection in bovine milk and serum samples was validated. The assay use B. abortus smooth lipopolysaccharide as antigen, immobilized on a polystyrene matrix; milk diluted 1:2 or serum diluted 1:50, in a buffer containing divalent cation chelating agents EDTA and EGTA (ethyleneglycol-bis-aminoether-N,N,N',N'-tetraacetic acid) to reduce non-specific reactions; and a mouse monoclonal antibody specific for an epitope of bovine IgG1, conjugated with horseradish peroxidase. A total of 2646 sera and 2119 milk samples from cows older than 24 months were obtained from 12 brucellosis-free herds for at least the previous 5 years. Milk samples were obtained in parallel with serum samples. The remaining 527 serum samples were from dry cows. All cattle were vaccinated with B. abortus strain 19 between 3-10 months of age. Five hundred and fifty-two milk samples and 562 serum samples were obtained from 6 infected herds with abortions where B. abortus was isolated at least once no more than 6 months before sampling. The complement-fixation test (CFT) on serum samples was considered the gold standard. Serum samples were also tested with the official screening test: the buffered plate antigen (BPA) test. The cut-off point was determined using receiver-operating characteristic (ROC) analysis. For milk samples, it was fixed at 36 percent positivity (PP) giving a sensitivity of 99.6% with a 95% confidence interval (CI) of 98.6-99.9%. The specificity was 99.1% (CI 98.9-99.4%). For serum samples, the cut-off was fixed at 53 PP giving a sensitivity of 99.6% (CI 98.6-99.9%) and a specificity 98.6% (CI 98-99%). The BPA test showed a relative sensitivity of 99.6% (CI 98.6-99.9%) and a relative specificity of 98.6% (CI 98.1-99%). Our results indicate that the indirect ELISA is a highly sensitive and specific test and can be adapted to process a large number of samples.

  3. Serum Biochemistry of Lumpy Skin Disease Virus-Infected Cattle

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    Murat Şevik

    2016-01-01

    Full Text Available Lumpy skin disease is an economically important poxvirus disease of cattle. Vaccination is the main method of control but sporadic outbreaks have been reported in Turkey. This study was carried out to determine the changes in serum biochemical values of cattle naturally infected with lumpy skin disease virus (LSDV. For this study, blood samples in EDTA, serum samples, and nodular skin lesions were obtained from clinically infected animals (n=15 whereas blood samples in EDTA and serum samples were collected from healthy animals (n=15. A quantitative real-time PCR method was used to detect Capripoxvirus (CaPV DNA in clinical samples. A real-time PCR high-resolution melt assay was performed to genotype CaPVs. Serum cardiac, hepatic, and renal damage markers and lipid metabolism products were measured by autoanalyzer. LSDV nucleic acid was detected in all samples which were obtained from clinically infected cattle. The results of serum biochemical analysis showed that aspartate aminotransferase, alkaline phosphatase, total protein, and creatinine concentrations were markedly increased in serum from infected animals. However, there were no significant differences in the other biochemical parameters evaluated. The results of the current study suggest that liver and kidney failures occur during LSDV infection. These findings may help in developing effective treatment strategies in LSDV infection.

  4. Serum Biochemistry of Lumpy Skin Disease Virus-Infected Cattle.

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    Şevik, Murat; Avci, Oğuzhan; Doğan, Müge; İnce, Ömer Barış

    2016-01-01

    Lumpy skin disease is an economically important poxvirus disease of cattle. Vaccination is the main method of control but sporadic outbreaks have been reported in Turkey. This study was carried out to determine the changes in serum biochemical values of cattle naturally infected with lumpy skin disease virus (LSDV). For this study, blood samples in EDTA, serum samples, and nodular skin lesions were obtained from clinically infected animals (n = 15) whereas blood samples in EDTA and serum samples were collected from healthy animals (n = 15). A quantitative real-time PCR method was used to detect Capripoxvirus (CaPV) DNA in clinical samples. A real-time PCR high-resolution melt assay was performed to genotype CaPVs. Serum cardiac, hepatic, and renal damage markers and lipid metabolism products were measured by autoanalyzer. LSDV nucleic acid was detected in all samples which were obtained from clinically infected cattle. The results of serum biochemical analysis showed that aspartate aminotransferase, alkaline phosphatase, total protein, and creatinine concentrations were markedly increased in serum from infected animals. However, there were no significant differences in the other biochemical parameters evaluated. The results of the current study suggest that liver and kidney failures occur during LSDV infection. These findings may help in developing effective treatment strategies in LSDV infection.

  5. Serum antibody to neospora caninum in indigenous African cattle ...

    African Journals Online (AJOL)

    Sera from 78 indigenous cattle were tested, by the indirect fluorescent antibody technique (IFAT), for neosporosis. In vitro cultured Neospora caninum was used as antigen. Antibodies to Neospora at titres 1/640 and above were detected in two samples (2.6%), a titre considered diagnostic for the disease. All the other serum ...

  6. Serum biochemical parameters and cytokine profiles associated with natural African trypanosome infections in cattle.

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    Bakari, Soale Majeed; Ofori, Jennifer Afua; Kusi, Kwadwo Asamoah; Aning, George Kwame; Awandare, Gordon Akanzuwine; Carrington, Mark; Gwira, Theresa Manful

    2017-06-27

    Animal African trypanosomiasis (AAT) greatly affects livestock production in sub-Saharan Africa. In Ghana prevalence of AAT is estimated to range between 5 and 50%. Studies have reported serum biochemical aberrations and variability in cytokine profiles in animals during infection. However, information regarding the biochemical parameters and cytokine profiles associated with natural infections are limited. This study was therefore aimed at investigating changes in the levels of serum biochemical parameters and inflammatory cytokines during a natural infection. Nested internal transcribed spacer (ITS)-based PCR and sequencing were used to characterise trypanosome infection in cattle at two areas in Ghana (Adidome and Accra) of different endemicities. The cattle were sampled at four to five-week intervals over a period of six months. Levels of serum biochemical parameters, including creatinine, cholesterol, alkaline phosphatase (ALP), alanine aminotransferase (ALT), total bilirubin and total protein and cytokines (interleukin 10, interleukin 4, interleukin 12, interferon gamma and tumor necrosis factor alpha) were measured in serum samples and then compared between infected cattle and uninfected controls. The predominant trypanosome species detected in Accra (non-endemic) and Adidome (endemic) were Trypanosoma theileri and Trypanosoma vivax, respectively. Serum biochemical parameters were similar between infected and uninfected cattle in Accra. Infected cattle at Adidome however, had significantly higher levels of ALP, creatinine, total protein and total bilirubin (P biochemical alterations whereas cattle in a non-endemic area with predominantly chronic T. theileri infections demonstrate high anti-inflammatory response and no biochemical alterations.

  7. Hematological and serum biochemical findings in clinical cases of cattle naturally infected with lumpy skin disease.

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    Abutarbush, Sameeh M

    2015-03-15

    Lumpy skin disease (LSD) is an acute viral disease of cattle that is currently emerging in the Middle East region and poses a serious threat to Europe and the rest of the world. The objective of this study was to describe hematological and serum biochemical findings associated with natural clinical infection of LSD in cattle. A total of 129 animals clinically infected with LSD were enrolled in the study. Venous blood sample were collected from study animals, and hematological and serum biochemical parameters were measured. Leukocytopenia was found in 8.7%, while leucocytosis was found in 18.2% of affected cattle. Decreased hematocrit concentration was seen in 18.3%. Most affected cattle had reduced mean corpuscular volume (43.7%), mean corpuscular hemoglobin (14.3%), and mean corpuscular hemoglobin concentration (11.5%). All cattle with abnormal platelets count had thrombocytopenia. Hyperfibrinogenemia, hyperproteinemia, and hyperalbuminemia were found in 69%, 59.6%, and 37.2% of affected cattle, respectively. Decreased creatinine concentration was seen in 65.8%. Hyperkalemia and hyperchloremia was found in 9.6% and 10.4% of the affected cattle, respectively. LSD appears to be associated with inflammatory leukogram, anemia, thrombocytopenia, hyperfibrinogenemia, hyperproteinemia, decreased creatinine concentration, hyperchloramia, and hyperkalemia. These are likely due to the associated severe inflammatory process and disease complications such as anorexia and reduced muscle mass. This is the first study that documents hematological and serum biochemical findings associated with LSD infection. Understanding the blood profile picture may give further insight to the pathogenesis of the disease and help in treatment of individual cattle.

  8. Haemoglobin, serum albumin and transferrin variants of Bali (Banteng) cattle, Bos (Bibos) javanicus.

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    Bell, K; McKenzie, H A; Shaw, D C

    1990-01-01

    1. Individual blood samples from 144 Bali (Banteng) cattle [Bos (Bibos) javanicus] in the Northern Territory of Australia and from 61 Bali cross cattle, were examined by zone electrophoresis to determine the variants of haemoglobin, serum albumin and transferrin that are present. 2. Of the common cattle haemoglobin variants (A and B) only variant B occurs in the Bali cattle samples. A second variant, designated CBali, occurs in Bali cattle either as the heterozygote (B CBali) or as the homozygote, the frequencies of occurrence indicating a two-allele system of inheritance without dominance. The CBali cross samples may exhibit the homozygous or heterozygous A variant. 3. The CBali variant has an electrophoretic mobility intermediate between those of the A and B variants at pH 8.6 and 9.1 but closer to B than to A (B greater than C greater than A). It appears to be similar in mobility to the C variants found in Indian Khillan (CKhillan) by Naik, Sukumaran and Sanghvi (Anim. Prodn, 1965 I, 275-277), and in Asian cattle by Oishi, Abe and Namikama (Immunogenet. Lett., 1968 5, 170-173) and Abe, Mogi, Oishi, Tanaka and Suzuki (Proc. XIIth Europ. Conf. Anim. Blood Groups Biochem. Polymorphisms 1972, pp. 225-228), but appreciably different from those in Kenyan and Rhodesian cattle (CRhodesia) found by Braend (Anim. Blood Grps Biochem. Genet., 1971 2, 15-21) and Carr (Rhod. J. agric. Res., 1964 3, 62-62A), respectively. It is also different in mobility from the C variant found by Winter, Mayr, Schleger, Dworak, Krutzler and Burger (Res. vet. Sci., 1984 36, 276-283) in the mithun.(ABSTRACT TRUNCATED AT 250 WORDS)

  9. Reduced serum vitamin D concentrations in healthy early-lactation dairy cattle.

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    Holcombe, S J; Wisnieski, L; Gandy, J; Norby, B; Sordillo, L M

    2017-11-22

    Cattle obtain vitamin D by ingestion or cutaneous exposure to UV light. Dairy cattle diets are frequently supplemented with vitamin D to compensate for limited sun exposure or during times of increased metabolic demands, such as the periparturient period, to maintain calcium homeostasis. Whether housing and supplemental vitamin D practices supply adequate amounts of vitamin D to optimally support the transition from gestation to lactation in dairy cattle is unknown. Our objective was to determine how serum vitamin D concentrations of dairy cows change with season, age, parity, and stage of lactation. Clinically healthy cows (n = 183) from 5 commercial dairies were enrolled in the study. Serum samples were collected at dry off, within 7 d of entering the close-up group, and within 7 d after calving (calving+7). Vitamin D status was determined by measuring serum 25-hydroxyvitamin D [25(OH)D] by radioimmunoassay. We performed repeated-measures mixed-effects linear regression to determine the effects of season, age, parity, and lactation stage (dry off, close-up, and calving+7) on 25(OH)D concentrations in serum. Bivariable analysis indicated that parity, age, and season were not associated with serum 25(OH)D concentrations. Sample period affected 25(OH)D concentrations, with the highest 25(OH)D levels at dry off (99.7 ± 1.9 ng/mL) followed by close up (93.8 ± 2.1 ng/mL), with the lowest levels at calving+7 (82.6 ± 1.7 ng/mL). These data showed a large depletion of 25(OH)D in dairy cattle postpartum compared with late prepartum, although the biological significance of this change in these healthy cattle is unclear. Consumption of serum 25(OH)D by immune system functions and calcium homeostasis in early lactation likely caused the reduction in serum 25(OH)D concentrations after calving. These results suggest that determining whether serum 25(OH)D concentrations are associated with the incidence of transition period disease is an appropriate next step. Assessing the

  10. The serum concentrations of lupine alkaloids in orally-dosed Holstein cattle

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    Teratogenic alkaloid-containing Lupinus spp. cause significant losses to the cattle industry. Previous research has suggested that Holstein cattle clear toxic Delphinium alkaloids from their serum at a greater rate than beef cattle. The toxicokinetics of lupine alkaloids in Holsteins are not known...

  11. (1H-NMR spectroscopy revealed Mycobacterium tuberculosis caused abnormal serum metabolic profile of cattle.

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    Yingyu Chen

    Full Text Available To re-evaluate virulence of Mycobacterium tuberculosis (M. tb in cattle, we experimentally infected calves with M. tb andMycobacterium bovisvia intratracheal injection at a dose of 2.0×10(7 CFU and observed the animals for 33 weeks. The intradermal tuberculin test and IFN-γin vitro release assay showed that both M. tb and M. bovis induced similar responses. Immunohistochemical staining of pulmonary lymph nodes indicated that the antigen MPB83 of both M. tb and M. bovis were similarly distributed in the tissue samples. Histological examinations showed all of the infected groups exhibited neutrophil infiltration to similar extents. Although the infected cattle did not develop granulomatous inflammation, the metabolic profiles changed significantly, which were characterized by a change in energy production pathways and increased concentrations of N-acetyl glycoproteins. Glycolysis was induced in the infected cattle by decreased glucose and increased lactate content, and enhanced fatty acid β-oxidation was induced by decreased TG content, and decreased gluconeogenesis indicated by the decreased concentration of glucogenic and ketogenic amino acids promoted utilization of substances other than glucose as energy sources. In addition, an increase in acute phase reactive serum glycoproteins, together with neutrophil infiltration and increased of IL-1β production indicated an early inflammatory response before granuloma formation. In conclusion, this study indicated that both M. tb and M.bovis were virulent to cattle. Therefore, it is likely that cattle with M. tb infections would be critical to tuberculosis transmission from cattle to humans. Nuclear magnetic resonance was demonstrated to be an efficient method to systematically evaluate M. tb and M. bovi sinfection in cattle.

  12. Circulating antigen levels in serum of cattle naturally infected with Schistosoma mattheei.

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    De Bont, J; Van Lieshout, L; Deelder, A M; Ysebaert, M T; Vercruysse, J

    1996-11-01

    Levels of 2 Schistosoma circulating antigens, the circulating anodic antigen (CAA) and the circulating cathodic antigen (CCA), were determined in serum samples collected, on a monthly basis over a period of 1.5 years, from 32 farm animals of different ages and from 12 tracer calves exposed to Schistosoma mattheei infection on a Zambian farm. Faecal egg counts were monitored in all animals and worm burdens in tracers determined after perfusion. Antigen determination tests in serum, with sensitivities between 95 and 100% in heifers and adult cows, proved to be excellent tools for the diagnosis of cattle schistosomiasis. Also in young calves, some infections could be demonstrated earlier by CCA determination than by faecal egg examination. A poor correlation was seen between the data for faecal egg counts and for CAA and CCA levels. It therefore appears that circulating antigen measurements in serum are of limited value as indicators of the pathogenesis of infection in cattle. Although all tracer calves were found infected at perfusion, large variations were recorded in antigen levels. An unexpected finding was the observation in farm animals of a clear seasonal pattern in CAA levels, with significant increase between August and October during the second half of the dry season, when animals are subjected to heavy physical and nutritional stress. It therefore appears that, although circulating antigen determination may provide an indication of the worm burden in ageing infections, possible variations of antigen clearance rate with the physiological condition of the host may complicate the interpretation of the results.

  13. METABOLITE CHARACTERIZATION IN SERUM SAMPLES FROM ...

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    take advantage of larger chemical shift spread of 13C resonances allowing a more detailed identification of ... fingerprints of various metabolites of serum samples of normal healthy control have been obtained which can ... fasting 10 mL of blood sample from each individual was taken and was allowed to clot in plastic.

  14. METABOLITE CHARACTERIZATION IN SERUM SAMPLES FROM ...

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    Metabolites, the end products of cellular process reflect the system level biological stress response. Hence, any enzymatic ... gradient HSQC adiabatic pulses. The experiments were performed in ..... Expansion of 1H-13C HSQC spectra of serum control healthy samples highlighting the resonance assignments in the region ...

  15. Comparison of the serum toxicokinetics of larkspur toxins in cattle, sheep and goats

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    Larkspurs (Delphinium spp.) are a major cause of cattle losses in western North America, whereas sheep are thought to be resistant to larkspur toxicosis. Goats are often used as a small ruminant model to study poisonous plants. In this study, we compared the serum toxicokinetic profile of toxic lark...

  16. Multiplex ready flow cytometric immunoassay for total insulin like growth factor 1 in serum of cattle

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    Bremer, M.G.E.G.; Smits, N.G.E.; Haasnoot, W.; Nielen, M.W.F.

    2010-01-01

    The European Union has banned the use of recombinant bovine somatotropins (rbST, growth hormones) to increase milk yield in dairy cattle. As direct detection of rbST in serum is problematic, methods based on the detection of changes in multiple rbST-dependent biomarkers have high potential for

  17. Measurement of trace elements in liver biopsy samples from cattle

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    Ouweltjes, W.; Zeeuw, de A.C.; Moen, A.; Counotte, G.H.M.

    2007-01-01

    Serum, plasma, or urine samples are usually used for the measurement of the trace elements copper, zinc, iron, selenium, because these samples are easy to obtain; however, these samples are not always appropriate. For example, it is not possible to measure molybdenum, the major antagonist of copper,

  18. Refractometer assessment of colostral and serum IgG and milk total solids concentrations in dairy cattle

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    2014-01-01

    Background Estimation of the quantity of colostral IgG or serum IgG absorbed following ingestion of colostrum by calves is essential for monitoring the effectiveness of colostrum feeding practices on dairy farms. Milk total solids concentrations determination is a critical part of quality assessment of nonsaleable whole milk prior to feeding to calves. To date, on-farm methods to assess colostral IgG, serum IgG or milk total solids concentrations have been performed separately with various instruments. The objective of this study was to evaluate the diagnostic performance of a single electronic, hand-held refractometer for assessing colostral and serum IgG concentrations and milk total solids in dairy cattle. Colostral IgG, serum IgG and milk total solids concentrations were determined by the refractometer. Corresponding analysis of colostral and serum IgG concentrations were determined by radial immunodiffusion (RID) while milk total solids were determined by spectrophotometry. Sensitivity and specificity of the refractometer for colostrum and serum samples were calculated as determined by RID. Sensitivity and specificity of the refractometer for milk samples was calculated as determined by spectrophotometry. Results The sensitivity of the refractometer was 1 for colostral IgG, serum IgG and milk total solids determinations. Specificity of the refractometer was 0.66, 0.24 and 0 for colostral IgG, serum IgG and milk total solids determinations, respectively. The refractometer underestimated colostral IgG, serum IgG and milk total solids concentrations compared to the concentrations determined by RID or spectrophotometry. Conclusions The refractometer was an acceptable, rapid, convenient on-farm method for determining colostral IgG and milk total solids. The refractometer was not an acceptable method for determination of serum IgG concentrations as it severely underestimated the serum IgG concentrations. PMID:25125217

  19. Seasonal evaluation of serumal Ca, P and ALP of slaughtered cattle in Tabriz abattoir

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    A.P Rezaei-Saber

    2008-08-01

    Full Text Available Increasing knowledge of metabolic disorders and prevention of any prduction decrease is beneficial in animal breeding. Problems related to calcium and phosphorus deficiency in high producing animals are among the most important metabolic disorders with disease such as osteomalacia, rickets, milk fever, pos parturition hemoglobinuria and reproductive disorders named as the metabolic complications. This study was conducted on non pregnant crossbreed dairy cattle. In the middle of each season with daily referral to the abattoir, blood samples were collected in veneject tubes and after transfer to the laboratory, the serumic levels of Ca, P and ALP were measured by spectrophotometry. One hundred samples for each season and a total of 400 samples were studied. The mean serumic levels of Ca, P and ALP were 8.83±0.02 mg/dl, 3.88±0.07 mg/dl, 163.4±0.8 Iu/lit; 9.05±0.03mg/dl, 4.19±0.01mg/dl, 240.7±1.2 Iu/lit; 9.03±0.07 mg/dl, 5.33±0.02mg/dl, 185.06±2.3 Iu/lit and 9.33±0.09 mg/dl, 5.74±0.08 mg/dl, 230.03±7.4 Iu/lit, in the spring, summer, autumn and winter respectively. Considering the normal Ca, P and ALP values in cattle, 6.3% and 83% of cases had Ca and P deficiency respectively in the spring, 80% of cases had subclinical phosphorus deficiency in the summer, 7.3% of cases had subclinical phosphorus deficiency in the autumn and 9.45% and 7.36% of cases had Ca deficiency and ALP elevation respectively in the winter. The significant decrease in Ca and P levels in the spring and P levels in the summer (p

  20. Metais pesados em amostras biológicas de bovinos Heavy metals in cattle biological samples

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    Maria Verônica de Souza

    2009-09-01

    Full Text Available O objetivo deste trabalho foi determinar a concentração de metais pesados no sangue (Pb, Ni e Cd, soro (Cu e Zn, pelo e leite (Pb, Ni, Cd, Cu e Zn de bovinos criados em área industrializada (com siderúrgicas e não-industrial do Estado de Minas Gerais, em amostras coletadas em duas épocas (inverno e verão, buscando avaliar a contaminação em animais em função do ambiente de exposição e da estação do ano. O local de criação dos animais afetou significativamente somente a concentração de Cu obtida nas amostras de soro, com maiores valores determinados no grupo de bovinos da região industrializada. A época de amostragem afetou a concentração dos metais Cu (soro, Zn (soro e leite, Pb (sangue e Cd (sangue e pelo, com as determinações efetuadas no verão proporcionando maiores teores do que as executadas no inverno, à exceção do Cd avaliado no pelo. Interações significativas (PThe aim of this research was to determine the heavy metals concentration in blood (Pb, Ni and Cd, serum (Cu and Zn, hair and milk (Pb, Ni, Cd, Cu and Zn of cattle raised in industrial (with steel mill and non-industrial areas in Minas Gerais, Brazil. The samples were collected during summer and winter, aiming to verify animals contamination related to environment and year season. The environment significantly influenced the concentration of Cu obtained on serum samples, with higher values for cattle from the industrialized area. The sampling time affected the concentration of Cu (serum, Zn (serum and milk, Pb (blood and Cd (blood and hair, with higher values for summer, except for Cd measured on hair. Meaningful interactions (P<0.05 between environment and year season were identified for Cu (hair and milk, Zn (hair and Ni (serum, hair and milk. The results obtained show that the presence of steel mills in a determined area does not mean, necessarily that higher concentration of heavy metals will be found in cattle biological matrices. The seasonality

  1. Detection of pregnancy by radioimmunoassay of a pregnancy serum protein (PSP60) in cattle

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    Mialon, M.M.; Renand, G.; Camous, S.; Martal, J.; Menissier, F. (Institut National de Recherches Agronomiques (INRA), 78 - Jouy-en-Josas (France))

    1994-01-01

    The accuracy and efficiency of pregnancy diagnoses in cattle by pregnancy serum protein (PSP60) radioimmunoassay, a progesterone radioimmunoassay or oestrus detection were compared. Blood samples were taken from 349 suckling heifers and cows ( 1 191 inseminations) at 28, 35, 50 and 90 d post-insemination for PSP60 determination and at 22-23 d for progesterone. Females were declared nonpregnant when plasma PSP60 concentration was lower than 0.2 ng/ml at 28, 35 and 50 d and 0.5 ng/ml at 90 d. When compared with rectal palpation at 90 d, the accuracy of positive (negative) diagnoses by progesterone assay was 80% (100%) in heifers and 75% (99%) in cows. The accuracy of positive diagnoses by PSP60 assay increased with gestation stage from 90% on d 28 in heifers (74% in cows) to 100% (99% in cows) at the time of rectal palpation. This accuracy was 84% on d 28 in cows when the interval from calving to blood sampling was higher than 115 d. Whatever the stage, the accuracy of negative diagnoses was higher than 90%. Efficiency in detecting pregnant or nonpregnant females on d 28 was equivalent to the progesterone assay. The method for detecting oestrus applied in this experiment was as efficient as the PSP60 or progesterone test at any stage of gestation. The PSP60 test is very flexible, which makes its use particularly interesting in naturally mated suckling herds because of the uncertainty regarding the date of fertilization. (authors).

  2. Serum haptoglobin concentrations in dairy cattle with lameness due to claw disorders.

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    Smith, Billy I; Kauffold, Johannes; Sherman, Lisa

    2010-11-01

    In cattle, elevated blood serum concentrations of haptoglobin, an acute phase protein, have been demonstrated in association with several diseases, but not with lameness. Serum haptoglobin was measured in 60 Holstein dairy cattle diagnosed with lameness due to four claw disorders, pododermatitis septica (PS; n=41), pododermatitis circumscripta (PC; n=8), interdigital necrobacillosis (IN; n=7), papillomatous digital dermatitis (PDD; n=4). Haptoglobin was measured on day 1 (0-3 days after lameness was observed but before treatment) and on days 3 and 5. A total of 10 healthy cows served as controls (haptoglobin values <1.0 mg/dL). Each of the claw disorders was associated with elevated haptoglobin on day 1 (PS, PC, IN and PDD: 65.9%, 37.5%, 71.4% and 25.0%, respectively). Trimming and antibiotic treatment led to a reduction in the number of PS and IN cows with increased haptoglobin concentrations, respectively (P<0.05), but trimming did not lead to any reduction in cows with PC. The study showed that lameness due to claw disorders can be associated with a systemic acute phase response and elevated serum haptoglobin in dairy cattle. Based on the course of haptoglobin, treatments seemed effective for all claw disorders except for PC. Published by Elsevier Ltd.

  3. Season and location effects on serum and liver mineral concentrations of Senepol cattle on St Croix, Virgin Islands.

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    Wildeus, S; McDowell, L R; Fugle, J R

    1992-11-01

    Serum and liver concentrations of selected macro- and trace minerals were determined in Senepol cattle at 8 sites (4 each in a high and low rainfall region) during the dry and wet season on St Croix. At each site an average of 15 mature, lactating cows, grazing native grass/legume pastures without supplementation were blood sampled each season. Liver samples were collected (n = 51) at slaughter from mature animals originating from the same sites. A preliminary analysis indicated no differences in serum mineral concentrations between mature lactating cows and growing heifers. There were differences between sites for serum magnesium (Mg) (P < 0.001), copper (Cu) (P < 0.05) selenium (Se) (P < 0.001) and zinc (Zn) (P < 0.01) in the dry season, and for Cu (P < 0.01), iron (Fe) (P < 0.001) and Zn (P < 0.01) in the wet season. Higher (P < 0.001) serum concentrations of Mg, Cu, Fe and Zn were observed in the dry season, while Se was higher (P < 0.01) in the wet season. Liver concentrations of Cu and Fe were lower (P < 0.01) and liver molybdenum (Mo) (P < 0.001) and Se (P < 0.05) higher during the dry season. The seasonal differences in serum Cu, Se and Zn concentrations have not been observed in other studies in the Central American region. More than 50% of serum samples were deficient in phosphorus (P) regardless of season, and in Cu and Zn during the wet season. Mineral supplementation should be considered.

  4. Establishing presence of antibodies against bovine respiratory syncytial virus (BRSV, parainfluenza virus 3 (PI3 and bovine herpesvirus 1 (BHV 1 in blood serum of cattle using indirect immunoenzyme probe

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    Šamanc Horea

    2009-01-01

    Full Text Available A total of 92 samples of bovine blood serum were examined for the presence of antibodies against the bovine respiratory syncytial virus using indirect immunoenzyme probe - iELISA. Specific antibodies against the bovine respiratory syncytial virus (BRSV were established in 46, or 50% blood serum samples. Investigations of the 92 blood serum samples of cattle for the presence of antibodies against the parainfluenza virus 3 (PI 3, revealed their presence in 77, or 83.69% of the samples, and the presence of antibodies against the bovine herpesvirus 1 (BHV 1 was established in 19, or 20.65% of the samples.

  5. Detection of serum neutralizing antibodies to Simbu sero-group viruses in cattle in Tanzania.

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    Mathew, Coletha; Klevar, S; Elbers, A R W; van der Poel, W H M; Kirkland, P D; Godfroid, J; Mdegela, R H; Mwamengele, G; Stokstad, M

    2015-08-15

    Orthobunyaviruses belonging to the Simbu sero-group occur worldwide, including the newly recognized Schmallenberg virus (SBV) in Europe. These viruses cause congenital malformations and reproductive losses in ruminants. Information on the presence of these viruses in Africa is scarce and the origin of SBV is unknown. The aim of this study was to investigate the presence of antibodies against SBV and closely related viruses in cattle in Tanzania, and their possible association with reproductive disorders. In a cross-sectional study, serum from 659 cattle from 202 herds collected in 2012/2013 were analyzed using a commercial kit for SBV ELISA, and 61 % were positive. Univariable logistic regression revealed significant association between ELISA seropositivity and reproductive disorders (OR = 1.9). Sera from the same area collected in 2008/2009, before the SBV epidemic in Europe, were also tested and 71 (54.6 %) of 130 were positive. To interpret the ELISA results, SBV virus neutralization test (VNT) was performed on 110 sera collected in 2012/2013, of which 51 % were positive. Of 71 sera from 2008/2009, 21 % were positive. To investigate potential cross reactivity with related viruses, 45 sera from 2012/2013 that were positive in SBV ELISA were analyzed in VNTs for Aino, Akabane, Douglas, Peaton, Sabo, SBV, Sathuperi, Shamonda, Simbu and Tinaroo viruses. All 45 sera were positive for one or more of these viruses. Twenty-nine sera (64.4 %) were positive for SBV, and one had the highest titer for this virus. This is the first indication that Aino, Akabane, Douglas, Peaton, Sabo, SBV, Sathuperi, Shamonda and Tinaroo viruses circulate and cause negative effect on reproductive performance in cattle in Tanzania. SBV or a closely related virus was present before the European epidemic. However, potential cross reactivity complicates the interpretation of serological studies in areas where several related viruses may circulate. Virus isolation and molecular characterization

  6. Temperament Type Specific Metabolite Profiles of the Prefrontal Cortex and Serum in Cattle

    Science.gov (United States)

    Brand, Bodo; Hadlich, Frieder; Brandt, Bettina; Schauer, Nicolas; Graunke, Katharina L.; Langbein, Jan; Repsilber, Dirk; Ponsuksili, Siriluk; Schwerin, Manfred

    2015-01-01

    In the past decade the number of studies investigating temperament in farm animals has increased greatly because temperament has been shown not only to affect handling but also reproduction, health and economically important production traits. However, molecular pathways underlying temperament and molecular pathways linking temperament to production traits, health and reproduction have yet to be studied in full detail. Here we report the results of metabolite profiling of the prefrontal cortex and serum of cattle with distinct temperament types that were performed to further explore their molecular divergence in the response to the slaughter procedure and to identify new targets for further research of cattle temperament. By performing an untargeted comprehensive metabolite profiling, 627 and 1097 metabolite features comprising 235 and 328 metabolites could be detected in the prefrontal cortex and serum, respectively. In total, 54 prefrontal cortex and 51 serum metabolite features were indicated to have a high relevance in the classification of temperament types by a sparse partial least square discriminant analysis. A clear discrimination between fearful/neophobic-alert, interested-stressed, subdued/uninterested-calm and outgoing/neophilic-alert temperament types could be observed based on the abundance of the identified relevant prefrontal cortex and serum metabolites. Metabolites with high relevance in the classification of temperament types revealed that the main differences between temperament types in the response to the slaughter procedure were related to the abundance of glycerophospholipids, fatty acyls and sterol lipids. Differences in the abundance of metabolites related to C21 steroid metabolism and oxidative stress indicated that the differences in the metabolite profiles of the four extreme temperament types could be the result of a temperament type specific regulation of molecular pathways that are known to be involved in the stress and fear response

  7. Limited interlaboratory comparison of Schmallenberg virus antibody detection in serum samples.

    NARCIS (Netherlands)

    Poel, van der W.H.M.; Cay, B.; Zientara, S.; Steinbach, F.; Valarcher, J.F.; Botner, A.; Mars, M.H.; Hakze-van der Honing, van der R.W.; Schirrmeier, H.; Beer, M.

    2014-01-01

    Eight veterinary institutes in seven different countries in Europe participated in a limited interlaboratory comparison trial to evaluate laboratory performances of Schmallenberg virus (SBV) antibody detection in serum. Seven different sheep sera and three different cattle sera were circulated, and

  8. Dairy cattle serum and milk factors contributing to the risk of colon and breast cancers.

    Science.gov (United States)

    zur Hausen, Harald; de Villiers, Ethel-Michele

    2015-08-15

    The analysis of published epidemiological data on colon and breast cancer reveals a remarkable concordance for most regions of the world. A low incidence for both cancers has been recorded in Mongolia and Bolivia. Discrepant data, however, have been reported for India, Japan and Korea. In India, the incidence of breast cancer is significantly higher than for colon cancer, in Japan and Korea colon cancer exceeds by far the rate of breast cancer. Here, studies are summarized pointing to a species-specific risk for colon cancer after consumption of beef originating from dairy cattle. Uptake of dairy products of Bos taurus-derived milk cattle, particularly consumed at early age, is suggested to represent one of the main risk factors for the development of breast cancer. A recent demonstration of reduced breast cancer rates in individuals with lactose intolerance (Ji et al., Br J Cancer 2014; 112:149-52) seems to be in line with this interpretation. Species-specific risk factors for these cancers are compatible with the transmission of different infectious factors transferred via meat or dairy products. Countries with discordant rates of colon and breast cancer reveal a similar discordance between meat and milk product consumption of dairy cattle. The recent isolation of a larger number of novel presumably viral DNAs from serum, meat and dairy products of healthy dairy cows, at least part of them infectious for human cells, deserves further investigation. Systemic infections early in life, resulting in latency and prevention of subsequent infections with the same agent by neutralizing antibodies, would require reconsideration of ongoing prospective studies conducted in the adult population. © 2015 UICC.

  9. Evaluation of serum and milk ELISAs for paratuberculosis in Danish dairy cattle

    DEFF Research Database (Denmark)

    Klausen, Joan; Huda, A.; Ekeroth, Lars

    2003-01-01

    A milk and a serum ELISA for detection of antibodies against Mycobacterium avium ssp. paratuberculosis (MAP) were evaluated against the complement-fixation test (CFT) and culture of faecal samples from 580 cows collected between August 1996 and December 1996. Milk and serum were obtained...... concurrently from six dairy herds infected with MAP and from two dairy herds without history of infection with MAP. A cut-off value of 7 OD% was used in the ELISAs. At this cut-off value, all six culture-positive herds were positive in the serum ELISA but one was negative in the milk ELISA. All six culture......-positive herds were positive in the CFT. In the two culture-negative herds, the serum and the milk ELISA deemed all serum samples negative at this cut-off value, whereas four serum samples from one of these herds were positive in the CFT. The highest cut-off value enabling the milk ELISA to record all six...

  10. A comparison of serologic tests for the detection of serum antibodies to whole-cell and recombinant Borrelia burgdorferi antigens in cattle.

    Science.gov (United States)

    Magnarelli, Louis A; Bushmich, Sandra L; Sherman, Bruce A; Fikrig, Erol

    2004-08-01

    Serum samples from healthy dairy and beef cattle, living in tick-infested areas of Connecticut, USA, were analyzed by polyvalent enzyme-linked immunosorbent assays (ELISA), indirect fluorescent antibody (IFA) staining methods, or Western blot procedures to detect antibodies to tick-borne agents. Of the 80 sera tested by ELISA with whole-cell or 10 separate recombinant antigens (fusion proteins) of Borrelia burgdorferi sensu stricto, 57 (71%) were positive to 1 or more antigens, while 36 (45%) reacted to whole-cell antigens by IFA staining methods. Three (4%) of 80 samples had antibodies to Anaplasma phagocytophilum. There were antibodies to outer surface protein (Osp) A, OspB, OspC, OspE, OspF, protein (p) 41-G, p35, p37, and VlsE antigens of B. burgdorferi, but there was no reactivity to the p39 antigen by ELISA. Western immunoblots of a subset of 9 sera verified antibody presence in all samples and showed distinct reactivities to multiple proteins having molecular masses of about 31 kilodaltons (kDa), 34 kDa, 35 kDa, 41 kDa, and 83/93 kDa. High specificity (97%) was noted when 16 cattle sera containing antibodies to Leptospira interrogans serovars, Brucella sp., Anaplasma marginale, or A. phagocytophilum were tested by ELISA with separate whole-cell or recombinant B. burgdorferi antigens. An ELISA and Western blot analyses can be used to confirm the exposure of cattle to B. burgdorferi.

  11. Serum Dried Samples to Detect Dengue Antibodies: A Field Study

    Directory of Open Access Journals (Sweden)

    Angelica Maldonado-Rodríguez

    2017-01-01

    Full Text Available Background. Dried blood and serum samples are useful resources for detecting antiviral antibodies. The conditions for elution of the sample need to be optimized for each disease. Dengue is a widespread disease in Mexico which requires continuous surveillance. In this study, we standardized and validated a protocol for the specific detection of dengue antibodies from dried serum spots (DSSs. Methods. Paired serum and DSS samples from 66 suspected cases of dengue were collected in a clinic in Veracruz, Mexico. Samples were sent to our laboratory, where the conditions for optimal elution of DSSs were established. The presence of anti-dengue antibodies was determined in the paired samples. Results. DSS elution conditions were standardized as follows: 1 h at 4°C in 200 µl of DNase-, RNase-, and protease-free PBS (1x. The optimal volume of DSS eluate to be used in the IgG assay was 40 µl. Sensitivity of 94%, specificity of 93.3%, and kappa concordance of 0.87 were obtained when comparing the antidengue reactivity between DSSs and serum samples. Conclusion. DSS samples are useful for detecting anti-dengue IgG antibodies in the field.

  12. Early Detection of Foot-And-Mouth Disease Virus from Infected Cattle Using A Dry Filter Air Sampling System.

    Science.gov (United States)

    Pacheco, J M; Brito, B; Hartwig, E; Smoliga, G R; Perez, A; Arzt, J; Rodriguez, L L

    2017-04-01

    Foot-and-mouth disease (FMD) is a highly contagious livestock disease of high economic impact. Early detection of FMD virus (FMDV) is fundamental for rapid outbreak control. Air sampling collection has been demonstrated as a useful technique for detection of FMDV RNA in infected animals, related to the aerogenous nature of the virus. In the current study, air from rooms housing individual (n = 17) or two groups (n = 4) of cattle experimentally infected with FDMV A24 Cruzeiro of different virulence levels was sampled to assess the feasibility of applying air sampling as a non-invasive, screening tool to identify sources of FMDV infection. Detection of FMDV RNA in air was compared with first detection of clinical signs and FMDV RNA levels in serum and oral fluid. FMDV RNA was detected in room air samples 1-3 days prior (seven animals) or on the same day (four animals) as the appearance of clinical signs in 11 of 12 individually housed cattle. Only in one case clinical signs preceded detection in air samples by one day. Overall, viral RNA in oral fluid or serum preceded detection in air samples by 1-2 days. Six individually housed animals inoculated with attenuated strains did not show clinical signs, but virus was detected in air in one of these cases 3 days prior to first detection in oral fluid. In groups of four cattle housed together, air detection always preceded appearance of clinical signs by 1-2 days and coincided more often with viral shedding in oral fluid than virus in blood. These data confirm that air sampling is an effective non-invasive screening method for detecting FMDV infection in confined to enclosed spaces (e.g. auction barns, milking parlours). This technology could be a useful tool as part of a surveillance strategy during FMD prevention, control or eradication efforts. Published 2015. This article is a U.S. Government work and is in the public domain in the USA.

  13. A radioimmunoassay detecting the bovine leukaemia virus transmembrane protein gp30 and anti-gp30 antibodies in the serum of cattle.

    Science.gov (United States)

    Bossmann, H; Siakkou, H; Ulrich, R; Uckert, W; Kraft, R; Rosenthal, S; Rosenthal, H A

    1989-03-01

    By means of SDS PAGE we isolated from virus-infected foetal lamb kidney (FLK) cells a relatively homogenous envelope transmembrane protein gp30 of bovine leukaemia virus (BLV). As shown by a partial sequence analysis of the N-terminus of this protein, our gp30 preparation contained only traces (less than 5%) of p24 gag protein: Rabbit anti-gp30 serum did not cross react with the BLV proteins gp51, p12, p15(1), p15(2), and p10 but reacted weakly with the p24 polypeptide. 125I-labelled gp30 (chloramine-T) was precipitated with the serum of BLV-infected cattle. Nonlabelled preparation of gp30 competitively inhibited the reaction of 125I-labelled gp30 with the natural antibodies. We investigated 193 cattle sera by liquid phase radioimmunoassays (RIA) using 125I-gp30, gp51 and p24 antigens. Sixteen noninfected cattle sera were negative in all tests. The 177 serum samples of BLV-infected animals were examined to the diagnostic value of the three tests. Of these, 175 were positive in gp51 RIA, 172 in p24 RIA and 164 in gp30 RIA. In all three tests, 159 sera were positive while 18 sera, mostly coming from animals with normal leukocyte counts, were positive only either with gp51 or p24, or were double positive with either gp51/p24 or gp51/gp30. We conclude that the gp51 RIA is superior to both the gp30 and the p24 RIA and that the gp30 RIA will be useful for investigating the role of gp30 in virus pathogenicity.

  14. Investigation of unusual high serum indices for lipemia in clear serum samples on siemens analysers dimension.

    Science.gov (United States)

    Fliser, Eva; Jerkovic, Ksenija; Vidovic, Tanja; Gorenjak, Maksimiljan

    2012-01-01

    We present our work of monitoring 202 different patients with markedly elevated serum index for lipemia whereby serum samples were clear. We tried to clarify the cause of occurrence of these indices which were detected in the years 2006-2010 on Siemens Dimension analyzers. In samples with unusual lipemia index we measured the concentration of lipids (total cholesterol, triglycerides, HDL and LDL cholesterol, Lp(a), ApoA1, ApoB), total proteins and checked for possible interferents (rheumatoid factor, immunoglobulins). We performed serum protein and immuno- electrophoresis. We investigated the repeatability of unusual lipemia indices during the day and after different time periods and we compared them on four different analyzers (RXL Max, Vista, Hitachi 911 and former Olympus AU640). In 87% of 202 samples we found a monoclonal or biclonal peak in serum protein electrophoresis. Different types of paraproteins were confirmed with immunofixation electrophoresis. In the remaining 13%, polyclonal elevated concentrations of immunoglobulins were measured. Other parameters had no influence on appearing of these indices. The repeatability of indices was good during the first day of measurements (P values > 0.05) and markedly lower in the next days or after 3 and 12 months (P values lipemia index in a clear serum sample measured on Siemens analyzers Dimension indicates a high possibility for the presence of a paraprotein in the sample.

  15. Utilization of composite fecal samples for detection of anthelmintic resistance in gastrointestinal nematodes of cattle.

    Science.gov (United States)

    George, Melissa M; Paras, Kelsey L; Howell, Sue B; Kaplan, Ray M

    2017-06-15

    Recent reports indicate that anthelmintic resistance in gastrointestinal nematodes of cattle is becoming increasingly prevalent worldwide. Presently, the fecal egg count reduction test (FECRT) is the only means available for detection of resistance to anthelmintics in cattle herds at the farm level. However, the FECRT is labor and cost intensive, and consequently is only rarely performed on cattle farms unless for research purposes. If costs could be reduced, cattle producers might be more likely to pursue drug resistance testing on their farms. One approach to reducing the cost of the FECRT, is the use of composite fecal samples for performing fecal egg counts (FEC), rather than conducting FEC on fecal samples from 15 to 20 individual animals. In this study FECRT were performed on 14 groups of cattle using both individual and composite FEC methods To measure how well the results of composite sampling reproduce those of individual sampling, Lin's Concordance Correlation Coefficient was utilized to describe both the linear relationship between methods and the slope and y-intercept of the line relating the data sets. There was little difference between the approaches with 98% agreement in mean FEC found between methods Mean FEC based on individual counts ranged between 0 and 670.6 eggs per gram of feces, indicating that the results of this study are applicable to a wide range of FEC levels. Standard error of the mean FEC and range of FEC are reported for each group prior to and following treatment to describe the variability of the data set. There was greater than 95% agreement in drug efficacy between individual and composite sampling methods, demonstrating composite sampling is appropriate to evaluate drug efficacy. Notably, for all groups tested the efficacy calculated by composite sampling was within the 95% confidence interval for efficacy calculated using individual sampling. The use of composite samples was shown to reduce the number of FEC required by 79

  16. Antibiotic resistance profile of bacteria isolated from raw milk samples of cattle and buffaloes

    Directory of Open Access Journals (Sweden)

    Tahlina Tanzin

    2016-03-01

    Conclusion: Two different species of bacteria i.e., S. aureus and E. coli are contaminating with milk samples. The pathogenic bacteria can be controlled effectively by using Ciprofloxacin and Levofloxacin in the case of mastitis in cattle and buffaloes in Bangladesh. [J Adv Vet Anim Res 2016; 3(1.000: 62-67

  17. The comparative utility of oral swabs and probang samples for detection of foot-and-mouth disease virus infection in cattle and pigs.

    Science.gov (United States)

    Stenfeldt, Carolina; Lohse, Louise; Belsham, Graham J

    2013-03-23

    Foot-and-mouth disease virus (FMDV) RNA was measured using quantitative reverse transcription-PCR (qRT-PCR) assays in oral swab and probang samples collected from cattle and pigs during experimental infections with serotype O FMDV. During acute infection, FMDV RNA was measurable in oral swabs as well as in probang samples from both species. FMDV RNA could be detected in oral swabs and probang samples from a time point corresponding to the onset of viremia in directly inoculated animals, whereas animals which were infected through contact exposure had low levels of FMDV RNA in oral swabs before viral RNA could be measured in serum. Analysis of samples collected from cattle persistently infected with FMDV showed that it was not possible to detect FMDV RNA in oral swabs harvested beyond 10 days post infection (dpi), despite the presence of FMDV RNA in probang samples that had been collected as late as 35 dpi. An interesting feature of the persistent infection in the cattle was the apparent decline in the level of FMDV RNA in probang samples after the acute phase of infection, which was followed by a marked rise again (in all the carrier animals) by 28 dpi. Results from this study indicate that qRT-PCR analysis of oral swabs is a useful approach in order to achieve a time efficient and reliable initial diagnosis of acute FMD in cattle and pigs, whereas probang sampling is essential for the detection of cattle that are persistently infected "carriers" of FMDV. Copyright © 2012 Elsevier B.V. All rights reserved.

  18. Substitution of Wheat for Corn in Beef Cattle Diets: Digestibility, Digestive Enzyme Activities, Serum Metabolite Contents and Ruminal Fermentation

    Science.gov (United States)

    Liu, Y. F.; Zhao, H. B.; Liu, X. M.; You, W.; Cheng, H. J.; Wan, F. C.; Liu, G. F.; Tan, X. W.; Song, E. L.; Zhang, X. L.

    2016-01-01

    The objective of this study was to evaluate the effect of diets containing different amounts of wheat, as a partial or whole substitute for corn, on digestibility, digestive enzyme activities, serum metabolite contents and ruminal fermentation in beef cattle. Four Limousin×LuXi crossbred cattle with a body weight (400±10 kg), fitted with permanent ruminal, proximal duodenal and terminal ileal cannulas, were used in a 4×4 Latin square design with four treatments: Control (100% corn), 33% wheat (33% substitution for corn), 67% wheat (67% substitution for corn), and 100% wheat (100% substitution for corn) on a dry matter basis. The results showed that replacing corn with increasing amounts of wheat increased the apparent digestibility values of dry matter, organic matter, and crude protein (pdigestibility of acid detergent fiber and neutral detergent fiber were lower with increasing amounts of wheat. Digestive enzyme activities of lipase, protease and amylase in the duodenum were higher with increasing wheat amounts (p<0.05), and showed similar results to those for the enzymes in the ileum except for amylase. Increased substitution of wheat for corn increased the serum alanine aminotransferase concentration (p<0.05). Ruminal pH was not different between those given only corn and those given 33% wheat. Increasing the substitution of wheat for corn increased the molar proportion of acetate and tended to increase the acetate-to-propionate ratio. Cattle fed 100% wheat tended to have the lowest ruminal NH3-N concentration compared with control (p<0.05), whereas no differences were observed among the cattle fed 33% and 67% wheat. These findings indicate that wheat can be effectively used to replace corn in moderate amounts to meet the energy and fiber requirements of beef cattle. PMID:26954111

  19. Cervical cancer detection based on serum sample Raman spectroscopy.

    Science.gov (United States)

    González-Solís, José Luis; Martínez-Espinosa, Juan Carlos; Torres-González, Luis Adolfo; Aguilar-Lemarroy, Adriana; Jave-Suárez, Luis Felipe; Palomares-Anda, Pascual

    2014-05-01

    The use of Raman spectroscopy to analyze the biochemical composition of serum samples and hence distinguish between normal and cervical cancer serum samples was investigated. The serum samples were obtained from 19 patients who were clinically diagnosed with cervical cancer, 3 precancer, and 20 healthy volunteer controls. The imprint was put under an Olympus microscope, and around points were chosen for Raman measurement.All spectra were collected at a Horiba Jobin-Yvon LabRAM HR800 Raman Spectrometer with a laser of 830-nm wavelength and 17-mW power irradiation. Raw spectra were processed by carrying out baseline correction, smoothing, and normalization to remove noise, florescence, and shot noise and then analyzed using principal component analysis (PCA). The control serum spectrum showed the presence of higher amounts of carotenoids indicated by peaks at 1,002, 1,160, and 1,523 cm(-1)and intense peaks associated with protein components at 754, 853, 938, 1,002, 1,300-1,345, 1,447, 1,523, 1,550, 1,620, and 1,654 cm(-1). The Raman bands assigned to glutathione (446, 828, and 1,404 cm(-1)) and tryptophan (509, 1,208, 1,556, 1,603, and 1,620 cm(-1)) in cervical cancer were higher than those of control samples, suggesting that their presence may also play a role in cervical cancer. Furthermore, weak bands in the control samples attributed to tryptophan (545, 760, and 1,174 cm(-1)) and amide III (1,234-1,290 cm(-1)) seem to disappear and decrease in the cervical cancer samples, respectively. It is shown that the serum samples from patients with cervical cancer and from the control group can be discriminated with high sensitivity and specificity when the multivariate statistical methods of PCA is applied to Raman spectra. PCA allowed us to define the wavelength differences between the spectral bands of the control and cervical cancer groups by confirming that the main molecular differences among the control and cervical cancer samples were glutathione, tryptophan,

  20. Relationship between blood serum insulin-like growth factor I concentration and postweaning feed efficiency of crossbred cattle at three levels of dietary intake.

    Science.gov (United States)

    Stick, D A; Davis, M E; Loerch, S C; Simmen, R C

    1998-02-01

    Blood serum concentration of IGF-I was analyzed to determine its relationship with individual postweaning feed efficiency (gain/feed) of 36 crossbred steer calves fed at three levels of feed intake (n = 12 at each level). Diets consisted of a corn silage-based growing diet for 84 d followed by a 91% concentrate finishing diet for 56 d. Dietary intake levels were at 80, 90, or 100% of ad libitum. Diets were formulated to ensure equal daily intake of protein, vitamins, and minerals across intake treatment levels. Intake was measured daily; ADG, DMI, and feed efficiency were calculated at 28-d intervals, through d 140. Individual weights and serum samples were collected at the beginning of the study and at 28-d intervals thereafter. The IGF-I concentrations were determined with a RIA. Data were analyzed as a multivariate split-plot in time. Imposed dietary intake restrictions did not affect serum IGF-I concentration (P = .90) or individual feed efficiency (P = .36), even though the least squares means for IGF-I concentration tended to decrease and the feed efficiency means tended to increase under the restricted intake levels. Serum IGF-I concentration, ADG, and feed efficiency were affected (P Residual correlations between IGF-I concentrations at adjacent 28-d sampling times averaged .72. Diet intake level x sampling time interactions existed for ADG (P = .02) and feed efficiency (P residual correlations of .28 (P feed efficiency, respectively. Regression analysis indicated that a 1 ng/mL increase in serum IGF-I concentration was associated with a .00135 kg/d increase in ADG (P feed increase in feed efficiency (P = .04). These results support the hypothesis that serum IGF-I plays a role in growth and in efficiency of feed utilization in beef cattle.

  1. Circulating microRNAs in serum from cattle challenged with Bovine Viral Diarrhea Virus

    Science.gov (United States)

    Bovine viral diarrhea virus (BVDV) is an RNA virus that is often associated with respiratory disease in cattle. MicroRNAs have been proposed as indicators of exposure to respiratory pathogens. The objective of this study was to identify microRNAs in cattle that had been challenged with a non-cytopat...

  2. Evaluation of thymus morphology and serum cortisol concentration as indirect biomarkers to detect low-dose dexamethasone illegal treatment in beef cattle

    Directory of Open Access Journals (Sweden)

    Vascellari Marta

    2012-08-01

    Full Text Available Abstract Background Corticosteroids are illegally used in several countries as growth promoters in veal calves and beef cattle, either alone or in association with sex steroids and β-agonists, especially at low dosages and primarily through oral administration, in order to enhance carcasses and meat quality traits. The aim of the present study is to evaluate the reliability of the histological evaluation of the thymus, as well as the serum cortisol determination, in identifying beef cattle, treated with two different dexamethasone-based growth-promoting protocols and the application of different withdrawal times before slaughter. Results Our findings demonstrate that low dosages of dexamethasone (DXM, administered alone or in association with clenbuterol as growth promoter in beef cattle, induce morphologic changes in the thymus, resulting in increase fat infiltration with concurrent cortical atrophy and reduction of the cortex/medulla ratio (C/M. In fact, the C/M value was significantly lower in treated animals than in control ones, with both the protocols applied. The cut off value of 0.93 for the cortex/medulla ratio resulted to be highly effective to distinguish control and treated animals. The animals treated with DXM showed inhibition of cortisol secretion during the treatment period, as well as at the slaughterhouse, 3 days after treatment suspension. The animals treated with lower doses of DXM in association with clenbuterol, showed inhibition of cortisol secretion during the treatment period, but serum cortisol concentration was restored to physiological levels at slaughterhouse, 8 days after treatment suspension. Conclusions The histological evaluation of thymus morphology, and particularly of the C/M may represent a valuable and reproducible method applicable to large-scale screening programs, due to the easy sampling procedures at slaughterhouse, as well as time and cost-saving of the analysis. Serum cortisol determination could be

  3. Association of preweaning and weaning serum cortisol and metabolites with ADG and incidence of respiratory disease in beef cattle

    Science.gov (United States)

    The objectives of this experiment were to determine the association of circulating cortisol, lactate, and glucose early in life on ADG and incidences of bovine respiratory disease (BRD) in cattle. A blood sample was collected approximately 3 wk prior to weaning and at weaning from genetically diver...

  4. Spatio-temporal variation in prevalence of Rift Valley fever: a post-epidemic serum survey in cattle and wildlife in Kenya

    Directory of Open Access Journals (Sweden)

    Olivia Wesula Lwande

    2015-12-01

    Full Text Available Background: Rift Valley fever (RVF is a fatal arthropod-borne zoonotic disease of livestock and humans. Since the identification of RVF in Kenya in the 1930s, repeated epizootics and epidemics coinciding with El Niño events have occurred in several locations in Africa and Saudi Arabia, causing mass deaths of livestock and humans. RVF is of great interest worldwide because of its negative effect on international livestock trade and its potential to spread globally. The latter is due to the increasing incidence of extreme climatic phenomena caused by global warming, as well as to the increase in global trade and international travel. How RVF is maintained and sustained between epidemics and epizootics is not clearly understood, but it has been speculated that wildlife reservoirs and trans-ovarian transmission in the vector may be important. Several studies have examined the role of wildlife and livestock in isolation or in a limited geographical location within the one country over a short time (usually less than a year. In this study, we examined the seroprevalence of anti-RVF antibodies in cattle and several wildlife species from several locations in Kenya over an inter-epidemic period spanning up to 7 years. Methods: A serological survey of immunoglobulin G (IgG antibodies to RVF using competitive ELISA was undertaken on 297 serum samples from different wildlife species at various locations in Kenya. The samples were collected between 2008 and 2015. Serum was also collected in 2014 from 177 cattle from Ol Pejeta Conservancy; 113 of the cattle were in close contact with wildlife and the other 64 were kept separate from buffalo and large game by an electric fence. Results: The seroprevalence of RVF virus (RVFV antibody was 11.6% in wildlife species during the study period. Cattle that could come in contact with wildlife and large game were all negative for RVFV. The seroprevalence was relatively high in elephants, rhinoceros, and buffalo, but

  5. DNA extraction methods and multiple sampling to improve molecular diagnosis of Sarcocystis spp. in cattle hearts.

    Science.gov (United States)

    Bräunig, Patrícia; Portella, Luiza Pires; Cezar, Alfredo Skrebsky; Libardoni, Felipe; Sangioni, Luis Antonio; Vogel, Fernanda Silveira Flores; Gonçalves, Paulo Bayard Dias

    2016-10-01

    Molecular detection of Sarcocystis spp. in tissue samples can be useful for experimental and diagnostic purposes. However, the parasite spreads unevenly through tissues, forming tissue cysts, and the cystic wall is an obstacle in DNA extraction protocols. Therefore, adequate sampling and effective disruption of the cysts are essential to improve the accuracy of DNA detection by PCR. The aims of this study were to evaluate the suitability of four protocols for DNA extraction from cysts of Sarcocystis spp. present in bovine myocardium samples or after their harvest in phosphate-buffered saline (PBS) solution as well as determine the effects of single or multiple sampling on the accuracy of molecular diagnosis of sarcocystosis in cattle hearts. Cysts and myocardium samples from nine bovine hearts were randomly distributed to four DNA extraction protocols: kit, kit with modification, DNAzol, and cetyl-trimethyl ammonium bromide (CTAB). Samples were submitted to DNA extraction and PCR as replicates of each heart (simplicate, duplicate, and triplicate), and the probability of a true positive diagnostic was calculated. Among the protocols tested, the kit with modification was determined to be the most suitable for DNA extraction from cysts in PBS solution (92.6 % of DNA detection by PCR); DNAzol resulted in higher DNA detection frequency from bovine myocardium samples (48.1 %). Multiple sampling improved the molecular diagnosis of Sarcocystis spp. infection in cattle hearts, increasing at 22.2 % the rate of true positive diagnostic.

  6. Apparent Prevalence of Beef Carcasses Contaminated with Mycobacterium avium subsp. paratuberculosis Sampled from Danish Slaughter Cattle

    Directory of Open Access Journals (Sweden)

    Hisako Okura

    2011-01-01

    Full Text Available Presence of Mycobacterium avium subsp. paratuberculosis (MAP in beef has been reported as a public health concern because asymptomatically infected cattle may contain MAP in tissues that are used for human consumption. Associations between MAP carcasses contamination and animal characteristics such as age, breed, production type, and carcass classification were assessed. Cheek muscles from 501 carcasses were sampled cross-sectionally at a Danish abattoir and tested for presence of viable MAP and MAP DNA by bacterial culture and IS900 realtime PCR, respectively. Cheek muscle tissues from carcasses of two dairy cows were positive by culture whereas 4% of the animals were estimated with ≥10 CFU/gram muscle based on realtime PCR. Age was found to be associated with carcass contamination with MAP. The observed viable MAP prevalence in beef carcasses was low. However, detection of MAP and MAP DNA in muscle tissues suggested that bacteremia occurred in slaughtered cattle.

  7. Evaluation of Rectoanal Mucosal Swab Sampling for Molecular Detection of Enterohemorrhagic Escherichia coli in Beef Cattle.

    Science.gov (United States)

    Agga, Getahun E; Arthur, Terrance M; Hinkley, Susanne; Bosilevac, Joseph M

    2017-04-01

    Cattle are a primary reservoir of enterohemorrhagic Escherichia coli (EHEC), and contaminated beef products are a source of human infections. The U.S. Department of Agriculture Food Safety and Inspection Service declared seven EHEC serogroups (O26, O45, O103, O111, O121, O145, and O157) as adulterants in raw ground beef. Sampling a large number of animals for EHEC surveillance or evaluations of EHEC-focused preharvest interventions requires a convenient and robust sampling method. We evaluated the diagnostic performance of rectoanal mucosal swab (RAMS) for the detection of the top seven EHEC serogroups. Paired fecal grab (FG) and RAMS samples were collected from 176 beef cattle and tested using the NeoSEEK Shiga toxin-producing E. coli (STEC) confirmation method. The prevalence of virulence-associated genes (stx 1 , stx 2 , stx 2c , eae, and nleB) was higher in RAMS than in FG samples. The results of the two methods had poor agreement, as indicated by kappa statistics, for the detection of the seven serogroups. When FG and RAMS results were combined for comparison, RAMS was more sensitive than FG for the detection of serogroups O103 (82% versus 39%), O157 (75% versus 67%), and O45 (79% versus 73%) with similar sensitivity for the detection of serogroup O145 (67%). Serogroups O111 and O121 were detected from one and two samples, respectively, by FG and were not detected by RAMS. Serogroup O26 was not detected with either method. RAMS appears to be equivalent or superior to FG sampling for detection of the top seven EHEC serogroups in the feces of beef cattle with the NeoSEEK STEC confirmation test.

  8. Associations of rumen parameters with feed efficiency and sampling routine in beef cattle.

    Science.gov (United States)

    Lam, S; Munro, J C; Zhou, M; Guan, L L; Schenkel, F S; Steele, M A; Miller, S P; Montanholi, Y R

    2017-11-10

    Characterizing ruminal parameters in the context of sampling routine and feed efficiency is fundamental to understand the efficiency of feed utilization in the bovine. Therefore, we evaluated microbial and volatile fatty acid (VFA) profiles, rumen papillae epithelial and stratum corneum thickness and rumen pH (RpH) and temperature (RT) in feedlot cattle. In all, 48 cattle (32 steers plus 16 bulls), fed a high moisture corn and haylage-based ration, underwent a productive performance test to determine residual feed intake (RFI) using feed intake, growth, BW and composition traits. Rumen fluid was collected, then RpH and RT logger were inserted 5.5±1 days before slaughter. At slaughter, the logger was recovered and rumen fluid and rumen tissue were sampled. The relative daily time spent in specific RpH and RT ranges were determined. Polynomial regression analysis was used to characterize RpH and RT circadian patterns. Animals were divided into efficient and inefficient groups based on RFI to compare productive performance and ruminal parameters. Efficient animals consumed 1.8 kg/day less dry matter than inefficient cattle (P⩽0.05) while achieving the same productive performance (P⩾0.10). Ruminal bacteria population was higher (P⩽0.05) (7.6×1011 v. 4.3×1011 copy number of 16S rRNA gene/ml rumen fluid) and methanogen population was lower (P⩽0.05) (2.3×109 v. 4.9×109 copy number of 16S rRNA gene/ml rumen fluid) in efficient compared with inefficient cattle at slaughter with no differences (P⩾0.10) between samples collected on-farm. No differences (P⩾0.10) in rumen fluid VFA were also observed between feed efficiency groups either on-farm or at slaughter. However, increased (P⩽0.05) acetate, and decreased (P⩽0.05) propionate, butyrate, valerate and caproate concentrations were observed at slaughter compared with on-farm. Efficient had increased (P⩽0.05) rumen epithelium thickness (136 v. 126 µm) compared with inefficient cattle. Efficient animals

  9. Genotyping of Leptospira directly in urine samples of cattle demonstrates a diversity of species and strains in Brazil.

    Science.gov (United States)

    Hamond, C; Pestana, C P; Medeiros, M A; Lilenbaum, W

    2016-01-01

    The aim of this study was to identify Leptospira in urine samples of cattle by direct sequencing of the secY gene. The validity of this approach was assessed using ten Leptospira strains obtained from cattle in Brazil and 77 DNA samples previously extracted from cattle urine, that were positive by PCR for the genus-specific lipL32 gene of Leptospira. Direct sequencing identified 24 (31·1%) interpretable secY sequences and these were identical to those obtained from direct DNA sequencing of the urine samples from which they were recovered. Phylogenetic analyses identified four species: L. interrogans, L. borgpetersenii, L. noguchii, and L. santarosai with the most prevalent genotypes being associated with L. borgpetersenii. While direct sequencing cannot, as yet, replace culturing of leptospires, it is a valid additional tool for epidemiological studies. An unexpected finding from this study was the genetic diversity of Leptospira infecting Brazilian cattle.

  10. Adsorption of serum calcium by plastic sample cups

    Science.gov (United States)

    Hall, R. A.; Whitehead, T. P.

    1970-01-01

    Sera left overnight in plastic AutoAnalyzer sample cups may give low calcium values; the effect is attributed to adsorption of calcium onto the walls of the vessel. The adsorption is brought about by a rise in the pH of the sera, and factors which promote the rise in pH increase the adsorption. This phenomenon is of practical importance because as much as 10% of the calcium in the serum may be adsorbed. Adsorption occurs particularly onto the walls of polystyrene cups, and when polypropylene cups were used the adsorption was reduced. The phenomenon cannot be evaluated or controlled by the use of control sera. In order to avoid the sampling error, serum for calcium analysis should be used fresh or stored at 4°C under conditions such that any change in pH is minimal. Sera should not be left to stand in AutoAnalyzer cups at room temperature for longer than three hours before analysis. PMID:5430421

  11. Spectrophotometric assay of creatinine in human serum sample

    Directory of Open Access Journals (Sweden)

    Avinash Krishnegowda

    2017-05-01

    Full Text Available A new spectrophotometric method for the analysis of creatinine concentration in human serum samples is developed. The method explores the oxidation of p-methylamino phenol sulfate (Metol in the presence of copper sulfate and creatinine which yields an intense violet colored species with maximum absorbance at 530 nm. The calibration graph of creatinine by fixed time assay ranged from 4.4 to 620 μM. Recovery of creatinine in human serum samples varied from 101% to 106%. Limit of detection and limit of quantification were 0.145 μM and 0.487 μM respectively. Sandell’s sensitivity was 0.112 μg cm−2 and molar absorptivity was 0.101 × 104 L mol−1 cm−1. Within day precision was 2.5–4.8% and day-to-day precision range was 3.2–7.8%. The robustness and ruggedness of the method expressed in RSD values ranged from 0.78% to 2.12% and 1.32% to 3.46% respectively, suggesting that the developed method was rugged. This method provides good sensitivity and is comparable to standard Jaffe’s method with comparatively less interference from foreign substances.

  12. Implementation of immunohistochemistry on frozen ear notch tissue samples in diagnosis of bovine viral diarrhea virus in persistently infected cattle.

    Science.gov (United States)

    Bedeković, Tomislav; Lemo, Nina; Lojkić, Ivana; Beck, Ana; Lojkić, Mirko; Madić, Josip

    2011-12-05

    Bovine viral diarrhea is a contagious disease of domestic and wild ruminants and one of the most economically important diseases in cattle. Bovine viral diarrhea virus belongs to the genus Pestivirus, within the family Flaviviridae. The identification and elimination of the persistently infected animals from herds is the initial step in the control and eradication programs. It is therefore necessary to have reliable methods for diagnosis of bovine viral diarrhea virus. One of those methods is immunohistochemistry. Immunohistochemistry on formalin fixed, paraffin embedded tissue is a routine technique in diagnosis of persistently infected cattle from ear notch tissue samples. However, such technique is inappropriate due to complicated tissue fixation process and it requires more days for preparation. On the contrary, immunohistochemistry on frozen tissue was usually applied on organs from dead animals. In this paper, for the first time, the imunohistochemistry on frozen ear notch tissue samples was described. Seventeen ear notch tissue samples were obtained during the period 2008-2009 from persistently infected cattle. Samples were fixed in liquid nitrogen and stored on -20°C until testing. Ear notch tissue samples from all persistently infected cattle showed positive results with good section quality and possibility to determinate type of infected cells. Although the number of samples was limited, this study indicated that immunohistochemistry on formalin fixed paraffin embedded tissue can be successfully replaced with immunohistochemistry on frozen ear notch tissue samples in diagnosis of persistently infected cattle.

  13. Implementation of immunohistochemistry on frozen ear notch tissue samples in diagnosis of bovine viral diarrhea virus in persistently infected cattle

    Directory of Open Access Journals (Sweden)

    Bedeković Tomislav

    2011-12-01

    Full Text Available Abstract Background Bovine viral diarrhea is a contagious disease of domestic and wild ruminants and one of the most economically important diseases in cattle. Bovine viral diarrhea virus belongs to the genus Pestivirus, within the family Flaviviridae. The identification and elimination of the persistently infected animals from herds is the initial step in the control and eradication programs. It is therefore necessary to have reliable methods for diagnosis of bovine viral diarrhea virus. One of those methods is immunohistochemistry. Immunohistochemistry on formalin fixed, paraffin embedded tissue is a routine technique in diagnosis of persistently infected cattle from ear notch tissue samples. However, such technique is inappropriate due to complicated tissue fixation process and it requires more days for preparation. On the contrary, immunohistochemistry on frozen tissue was usually applied on organs from dead animals. In this paper, for the first time, the imunohistochemistry on frozen ear notch tissue samples was described. Findings Seventeen ear notch tissue samples were obtained during the period 2008-2009 from persistently infected cattle. Samples were fixed in liquid nitrogen and stored on -20°C until testing. Ear notch tissue samples from all persistently infected cattle showed positive results with good section quality and possibility to determinate type of infected cells. Conclusions Although the number of samples was limited, this study indicated that immunohistochemistry on formalin fixed paraffin embedded tissue can be successfully replaced with immunohistochemistry on frozen ear notch tissue samples in diagnosis of persistently infected cattle.

  14. PCR detection of Campylobacter fetus subspecies venerealis in smegma samples collected from dairy cattle in Fars, Iran

    Directory of Open Access Journals (Sweden)

    Saeid Hosseinzadeh

    2014-12-01

    Full Text Available Bovine venereal campylobacteriosis, caused by Campylobacter fetus subsp. venerealis (Cfv, is regarded as one of the major threats to the cattle industry around the world. Abortion and infertility are two important reproductive problems in cows infected with C. fetus subsp. venerealis. Reports on the presence of Cfv are scarce in the cattle, in Iran. Therefore, the present study was designed to examine the presence of Cfv in the reproductive tract of dairy cattle either slaughtered in Shiraz abattoir or dairy herds with a history of infertility and abortion, and further to identify and differentiate this micro-organism in dairy cattle in Fars, south of Iran. A total of 95 smegma samples from the preputial cavity and the fornix of the cervical opening were collected using scraping method from bulls (n = 34 and cows (n = 61 in addition to eight samples of commercially bull frozen semen. Smegma samples were then cultured for isolation of Cfv and then the extracted DNA was examined for the presence of Cfv using an optimized multiplex PCR assay. None of the frozen semen samples examined were positive for Cfv. However, out of 95 smegma samples, thirteen animals (12.6% were found positive for Cfv consisting of 3 males and 10 females. In conclusion, the results of the current study clearly confirmed the presence of Cfv using PCR in the slaughtered cattle and dairy farms with a history of poor fertility and abortion in Fars, Iran.

  15. Studies On The Role Of Trade Cattle In The Transmission Of ...

    African Journals Online (AJOL)

    The role of trade cattle in the transmission of brucellosis was evaluated by determining the prevalence of Brucella antibodies in the traditional herd and in trade cattle. Rose Bengal Plate Test (RBPT), and serum agglutination test (SAT) were used to assay 162 and 56 serum samples collected from traditional herds and trade ...

  16. Analysis of the acute phase responses of Serum Amyloid A, Haptoglobin and Type 1 Interferon in cattle experimentally infected with foot-and-mouth disease virus serotype O

    DEFF Research Database (Denmark)

    Stenfeldt, Carolina; Heegaard, Peter M. H.; Stockmarr, Anders

    2011-01-01

    A series of challenge experiments were performed in order to investigate the acute phase responses to foot-and-mouth disease virus (FMDV) infection in cattle and possible implications for the development of persistently infected "carriers". The host response to infection was investigated through....... There was a statistically significant difference in the HP response between carriers and non-carriers with a lower response in the animals that subsequently developed into FMDV carriers. It was concluded that the induction of SAA, HP and type 1 IFN in serum can be used as markers of acute infection by FMDV in cattle....... measurements of the concentrations of the acute phase proteins (APPs) serum amyloid A (SAA) and haptoglobin (HP), as well as the bioactivity of type 1 interferon (IFN) in serum of infected animals. Results were based on measurements from a total of 36 infected animals of which 24 were kept for observational...

  17. Seroprevalence of Fasciola hepatica in cattle in Estonia

    DEFF Research Database (Denmark)

    Petersson, Jennifer; Jokelainen, Pikka; Lassen, Brian

    2017-01-01

    the seroprevalence and distribution of F. hepatica in cattle in Estonia. A total of 2461 individual serum samples from 218 farms distributed throughout all 15 Estonian counties, collected between February 2012 and March 2013, were tested for specific anti-F. hepatica antibodies using an in-house enzyme...... serological evidence of the presence and distribution of F. hepatica in cattle herds in Estonia....

  18. Apparent prevalence of beef carcasses contaminated with Mycobacterium avium subsp. paratuberculosis sampled from Danish slaughter cattle

    DEFF Research Database (Denmark)

    Okura, Hisako; Toft, Nils; Pozzato, Nicola

    2011-01-01

    of two dairy cows were positive by culture whereas 4% of the animals were estimated with =10¿CFU/gram muscle based on realtime PCR. Age was found to be associated with carcass contamination with MAP. The observed viable MAP prevalence in beef carcasses was low. However, detection of MAP and MAP DNA......Presence of Mycobacterium avium subsp. paratuberculosis (MAP) in beef has been reported as a public health concern because asymptomatically infected cattle may contain MAP in tissues that are used for human consumption. Associations between MAP carcasses contamination and animal characteristics...... such as age, breed, production type, and carcass classification were assessed. Cheek muscles from 501 carcasses were sampled cross-sectionally at a Danish abattoir and tested for presence of viable MAP and MAP DNA by bacterial culture and IS900 realtime PCR, respectively. Cheek muscle tissues from carcasses...

  19. Transplacental transfer of polycyclic aromatic hydrocarbons in paired samples of maternal serum, umbilical cord serum, and placenta in Shanghai, China.

    Science.gov (United States)

    Zhang, Xiaolan; Li, Xiaojing; Jing, Ye; Fang, Xiangming; Zhang, Xinyu; Lei, Bingli; Yu, Yingxin

    2017-03-01

    Prenatal exposure to polycyclic aromatic hydrocarbons (PAHs) is a high-priority public health concern. However, maternal to fetal transplacental transfer of PAHs has not been systematically studied. To investigate the transplacental transfer of PAHs from mother to fetus and determine the influence of lipophilicity (octanol-water partition coefficient, KOW) on transfer process, in the present study, we measured the concentrations of 15 PAHs in 95 paired maternal and umbilical cord serum, and placenta samples (in total 285 samples) collected in Shanghai, China. The average concentration of total PAHs was the highest in maternal serums (1290 ng g-1 lipid), followed by umbilical cord serums (1150 ng g-1 lipid). The value was the lowest in placenta samples (673 ng g-1 lipid). Low molecular weight PAHs were the predominant compounds in the three matrices. Increases in fish and meat consumption did not lead to increases in maternal PAH levels, and no obvious gender differences in umbilical cord serums were observed. The widespread presence of PAHs in umbilical cord serums indicated the occurrence of transplacental transfer. The ratios of PAH concentrations in umbilical cord serum to those in maternal serum (F/M) and the concentrations in placenta to those in maternal serum (P/M) of paired samples were analyzed to characterize the transfer process of individual PAHs. Most F/M ratios on lipid basis were close to one (range: 0.79 to 1.36), which suggested that passive diffusion may control the transplacental transfer of PAHs from maternal serum to the fetal circulation. The P/M and F/M values calculated on lipid basis showed that PAHs with lower KOW were more likely to transfer from mother to fetus via the placenta. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. EVALUATION OF ZEBU NELLORE CATTLE BLOOD SAMPLES USING THE CELL-DYN 3500 HEMATOLOGY ANALYZER

    Directory of Open Access Journals (Sweden)

    Alexandre Secorun Borges

    2014-12-01

    Full Text Available The Cell-dyn 3500 is a multiparameter flow cytometer, which may analyze samples from several species performing several simultaneous analyses. It is able to perform white blood cells, red blood cells and platelet counts, besides differential leukocyte counts, packed cell volume and hemoglobin determination. Cell-Dyn 3500 performs total leukocyte count both optically and by impedance. The equipment may choose one or other method, based on the reliability of the results. Erythrocyte and platelet counts are determined by impedance. Leukocyte differentiation is based on an optical principle, using separation in multiangular polarized light. The objective of this study was to compare the results of complete blood count of Zebu Nellore heifers from Celldyn 3500, with those obtained from a semi-automated cell counter (Celm CC 510 and the manual technique. Blood samples were collected from the jugular vein in 5 mL EDTA vacuum tubes from 58 Nellore heifers, at 24 months of age. Samples were processed in parallel in the three different techniques. Results were analyzed using paired t test, Pearson’s correlation and the Bland-Altmann method. There was a strong correlation for all parameters analyzed by Cell-Dyn 3500, manual method and semiautomated cell counter, except for basophils and monocytes counts. These results confirm that this analyzer is reliable for blood samples analysis of zebu cattle.

  1. Comparative proteomics dataset of skimmed milk samples from Holstein and Jersey dairy cattle

    Directory of Open Access Journals (Sweden)

    Rinske Tacoma

    2016-03-01

    Full Text Available Milk samples were collected from Holstein and Jersey breeds of dairy cattle maintained under the same management practices and environmental conditions over a seven-day period. Milk samples were collected twice daily from six cows of each breed as previously described (Tacoma et al., 2016 [1]. Samples were composited within individual cow over the experimental period and skimmed to remove the fat layer. Skimmed milk samples were fractionated using CaCl2 precipitation, ultracentrifugation and ProteoMiner treatment to remove the high abundance milk proteins. Separation of the low abundance proteins was achieved using SDS-PAGE. Differential protein abundances were analyzed by mass spectrometry-based proteomic approaches followed by statistical analyses of the peptide count data. The complete list of low-abundance proteins identified in both breeds is provided in the dataset as well as the total number of distinct sequenced peptides and gene ontology functions for each protein. The relative abundance of a select few proteins is depicted using the SIEVE software.

  2. Hemogram, serum biochemistry and hepatic histologic features in cattle after administration of citrus pulp

    OpenAIRE

    Oliveira,N.J.F.; Melo, M.M.; L.A. Lago; Nascimento, E.F.

    2005-01-01

    Hemogram and serum biochemistry (aspartate aminotransferase, alkaline phosphatase and gamma glutamiltransferase, total protein, urea, creatinine, calcium and phosphorus) were performed weekly in five crossbreed bovine after consumption of a diet containing citrus pulp pellets (40%), for 43 days. Percutaneous hepatic biopsy and histologic evaluation were performed in each animal before and after consumption of the citrus pulp diet. Hemogram, the enzymes aspartate aminotransferase and gamma glu...

  3. Development of a loop-mediated isothermal amplification assay for rapid, sensitive detection of Campylobacter jejuni in cattle farm samples.

    Science.gov (United States)

    Dong, Hee-Jin; Cho, Ae-Ri; Hahn, Tae-Wook; Cho, Seongbeom

    2014-09-01

    Campylobacter jejuni is a leading cause of bacterial foodborne disease worldwide. The detection of this organism in cattle and their environment is important for the control of C. jejuni transmission and the prevention of campylobacteriosis. Here, we describe the development of a rapid and sensitive method for the detection of C. jejuni in naturally contaminated cattle farm samples, based on real-time loop-mediated isothermal amplification (LAMP) of the hipO gene. The LAMP assay was specific (100% inclusivity and exclusivity for 84 C. jejuni and 41 non-C. jejuni strains, respectively), sensitive (detection limit of 100 fg/μl), and quantifiable (R(2) = 0.9133). The sensitivity of the LAMP assay was then evaluated for its application to the naturally contaminated cattle farm samples. C. jejuni strains were isolated from 51 (20.7%) of 246 cattle farm samples, and the presence of the hipO gene was tested using the LAMP assay. Amplification of the hipO gene by LAMP within 30 min (mean ~10.8 min) in all C. jejuni isolates (n = 51) demonstrated its rapidity and accuracy. Next, template DNA was prepared from a total of 186 enrichment broth cultures of cattle farm samples either by boiling or using a commercial kit, and the sensitivity of detection of C. jejuni was compared between the LAMP and PCR assays. In DNA samples prepared by boiling, the higher sensitivity of the LAMP assay (84.4%) compared with the PCR assay (35.5%) indicates that it is less susceptible to the existence of inhibitors in sample material. In DNA samples prepared using a commercial kit, both the LAMP and PCR assays showed 100% sensitivity. We anticipate that the use of this rapid, sensitive, and simple LAMP assay, which is the first of its kind for the identification and screening of C. jejuni in cattle farm samples, may play an important role in the prevention of C. jejuni contamination in the food chain, thereby reducing the risk of human campylobacteriosis.

  4. Effects of blood sample handling procedures on measurable inflammatory markers in plasma, serum and dried blood spot samples

    DEFF Research Database (Denmark)

    Skogstrand, K.; Thorsen, P.; Vogel, I.

    2008-01-01

    , and 30 days at the same temperatures. 27 inflammatory markers in serum and plasma and 25 markers in DBSS were measured by a previously validated multiplex sandwich immunoassay using Luminex xMAP technology. The measurable concentrations of several cytokines in serum and plasma were significantly......The interests in monitoring inflammation by immunoassay determination of blood inflammatory markers call for information on the stability of these markers in relation to the handling of blood samples. The increasing use of stored biobank samples for such ventures that may have been collected...... increased when blood samples were stored for a period of time before the centrifugation, for certain cytokines more than 1000 fold compared to serum and plasma isolated and frozen immediately after venepuncture. The concentrations in serum generally increased more than in plasma. The measurable...

  5. Seroprevalence of Neospora caninum Infection in Rural and Industrial Cattle in Northern Iran

    Directory of Open Access Journals (Sweden)

    MR Youssefi

    2009-02-01

    Full Text Available Background: Neospora caninum is an intracellular parasite which causes abortion in cattle worldwide.  The aim of this study was to determine the seroprevalence of N. caninum in cattle in Babol City, North of Iran."nMethods: Blood samples were collected from 237 cattle for determining the seroprevalence of N.  caninum.  A total of 237 serum samples were tested for anti-Neospora antibodies.  Serum samples were analyzed for antibodies against N. caninum antigen using a commercial N. caninum ELISA kit."nResults: Antibodies to N. caninum were found in 76 of 237 total cattle (32%, 40 of 155 industrial cat­tle (25. 8% and 36 of 82 rural cattle sera (43. 9% based on ELISA test results."nConclusion: This study is the first report of Neospora infection in this area.  Significant difference was observed regarding infection in industrial and rural cattle (P<0. 01.

  6. Post-death cloning of endangered Jeju black cattle (Korean native cattle): fertility and serum chemistry in a cloned bull and cow and their offspring.

    Science.gov (United States)

    Kim, Eun Young; Song, Dong Hwan; Park, Min Jee; Park, Hyo Young; Lee, Seung Eun; Choi, Hyun Yong; Moon, Jeremiah Jiman; Kim, Young Hoon; Mun, Seong Ho; Oh, Chang Eon; Ko, Moon Suck; Lee, Dong Sun; Riu, Key Zung; Park, Se Pill

    2013-12-17

    To preserve Jeju black cattle (JBC; endangered native Korean cattle), a pair of cattle, namely a post-death cloned JBC bull and cow, were produced by somatic cell nuclear transfer (SCNT) in a previous study. In the present study, we examined the in vitro fertilization and reproductive potentials of these post-death cloned animals. Sperm motility, in vitro fertilization and developmental capacity were examined in a post-death cloned bull (Heuk Oll Dolee) and an extinct nuclear donor bull (BK94-13). We assessed reproductive ability in another post-death cloned cow (Heuk Woo Sunee) using cloned sperm for artificial insemination (AI). There were no differences in sperm motility or developmental potential of in vitro fertilized embryos between the post-death cloned bull and its extinct nuclear donor bull; however, the embryo development ratio was slightly higher in the cloned sperm group than in the nuclear donor sperm group. After one attempt at AI, the post-death cloned JBC cow became pregnant, and gestation proceeded normally until day 287. From this post-death cloned sire and dam, a JBC male calf (Heuk Woo Dolee) was delivered naturally (weight, 25 kg). The genetic paternity/maternity of the cloned JBC bull and cow with regard to their offspring was confirmed using International Society for Animal Genetics standard microsatellite markers. Presently, Heuk Woo Dolee is 5 months of age and growing normally. In addition, there were no significant differences in blood chemistry among the post-death cloned JBC bull, the cow, their offspring and cattle bred by AI. This is the first report showing that a pair of cattle, namely, a post-death cloned JBC bull and cow, had normal fertility. Therefore, SCNT can be used effectively to increase the population of endangered JBC.

  7. Retrospective study of noroviruses in samples of diarrhoea from cattle, using the Veterinary Laboratories Agency's Farmfile database.

    Science.gov (United States)

    Milnes, A S; Binns, S H; Oliver, S L; Bridger, J C

    2007-03-10

    A collaborative study was undertaken by the Veterinary Laboratories Agency (vla) and the Royal Veterinary College (rvc) to determine the prevalence of bovine noroviruses in cattle with diarrhoea. Samples of bovine diarrhoea were provided by the vla from routine diagnostic submissions and a reverse transcription-pcr was used by the rvc to detect the viruses. Epidemiological information about the samples was provided retrospectively by the Farmfile database. Noroviruses were detected in 44 (11 per cent) of the 398 samples tested, and Farmfile data were used to investigate the differences between the positive and negative animals.

  8. Detection of occult hepatitis B in serum and oral fluid samples

    Directory of Open Access Journals (Sweden)

    Moyra Machado Portilho

    Full Text Available In occult hepatitis B infection (OBI, hepatitis B virus DNA (HBV DNA can be detected in serum samples; however, oral fluid collection for detection of HBV DNA has not yet been explored, despite the availability of collection devices. Serum and oral fluid samples from 45 hepatitis B core antibody (anti-HBc-positive patients were collected for the amplification of the HBV polymerase gene. HBV DNA was detected in five serum and four oral fluid samples (the detection limit for oral fluid was 1.656 log IU/mL in paired serum. In conclusion, simple methodologies of sample collection and in-house polymerase chain reaction (PCR allowed detection of HBV DNA, and these could be used to improve the diagnosis of OBI, especially in locations with limited resources.

  9. Clinical, radiological and molecular diagnosis correlation in serum samples from patients with osteoarticular tuberculosis

    Directory of Open Access Journals (Sweden)

    Guadalupe García-Elorriaga

    2014-07-01

    Conclusions: Nested PCR in serum samples is a rapid, highly sensitive and specific modality for OTB detection. PCR should be performed in addition to clinical evaluation, imaging studies, acid-fast bacilli staining, culture and histopathology diagnosis, if possible.

  10. Pre-analytical sample quality: metabolite ratios as an intrinsic marker for prolonged room temperature exposure of serum samples.

    Directory of Open Access Journals (Sweden)

    Gabriele Anton

    Full Text Available Advances in the "omics" field bring about the need for a high number of good quality samples. Many omics studies take advantage of biobanked samples to meet this need. Most of the laboratory errors occur in the pre-analytical phase. Therefore evidence-based standard operating procedures for the pre-analytical phase as well as markers to distinguish between 'good' and 'bad' quality samples taking into account the desired downstream analysis are urgently needed. We studied concentration changes of metabolites in serum samples due to pre-storage handling conditions as well as due to repeated freeze-thaw cycles. We collected fasting serum samples and subjected aliquots to up to four freeze-thaw cycles and to pre-storage handling delays of 12, 24 and 36 hours at room temperature (RT and on wet and dry ice. For each treated aliquot, we quantified 127 metabolites through a targeted metabolomics approach. We found a clear signature of degradation in samples kept at RT. Storage on wet ice led to less pronounced concentration changes. 24 metabolites showed significant concentration changes at RT. In 22 of these, changes were already visible after only 12 hours of storage delay. Especially pronounced were increases in lysophosphatidylcholines and decreases in phosphatidylcholines. We showed that the ratio between the concentrations of these molecule classes could serve as a measure to distinguish between 'good' and 'bad' quality samples in our study. In contrast, we found quite stable metabolite concentrations during up to four freeze-thaw cycles. We concluded that pre-analytical RT handling of serum samples should be strictly avoided and serum samples should always be handled on wet ice or in cooling devices after centrifugation. Moreover, serum samples should be frozen at or below -80°C as soon as possible after centrifugation.

  11. Pre-analytical sample quality: metabolite ratios as an intrinsic marker for prolonged room temperature exposure of serum samples.

    Science.gov (United States)

    Anton, Gabriele; Wilson, Rory; Yu, Zhong-Hao; Prehn, Cornelia; Zukunft, Sven; Adamski, Jerzy; Heier, Margit; Meisinger, Christa; Römisch-Margl, Werner; Wang-Sattler, Rui; Hveem, Kristian; Wolfenbuttel, Bruce; Peters, Annette; Kastenmüller, Gabi; Waldenberger, Melanie

    2015-01-01

    Advances in the "omics" field bring about the need for a high number of good quality samples. Many omics studies take advantage of biobanked samples to meet this need. Most of the laboratory errors occur in the pre-analytical phase. Therefore evidence-based standard operating procedures for the pre-analytical phase as well as markers to distinguish between 'good' and 'bad' quality samples taking into account the desired downstream analysis are urgently needed. We studied concentration changes of metabolites in serum samples due to pre-storage handling conditions as well as due to repeated freeze-thaw cycles. We collected fasting serum samples and subjected aliquots to up to four freeze-thaw cycles and to pre-storage handling delays of 12, 24 and 36 hours at room temperature (RT) and on wet and dry ice. For each treated aliquot, we quantified 127 metabolites through a targeted metabolomics approach. We found a clear signature of degradation in samples kept at RT. Storage on wet ice led to less pronounced concentration changes. 24 metabolites showed significant concentration changes at RT. In 22 of these, changes were already visible after only 12 hours of storage delay. Especially pronounced were increases in lysophosphatidylcholines and decreases in phosphatidylcholines. We showed that the ratio between the concentrations of these molecule classes could serve as a measure to distinguish between 'good' and 'bad' quality samples in our study. In contrast, we found quite stable metabolite concentrations during up to four freeze-thaw cycles. We concluded that pre-analytical RT handling of serum samples should be strictly avoided and serum samples should always be handled on wet ice or in cooling devices after centrifugation. Moreover, serum samples should be frozen at or below -80°C as soon as possible after centrifugation.

  12. Suitability of Frozen Serum Stored in Gel Separator Primary Sampling Tubes for Serological Testing

    OpenAIRE

    Rosa-Fraile, Manuel; Sampedro, Antonio; Rodríguez-Granger, Javier; Camacho, Enrique; Manrique, Ester

    2004-01-01

    The suitability of frozen serum after storage in primary sampling tubes with a gel separator for serological enzyme-linked immunosorbent assay testing (hepatitis B virus surface antigen [HBs Ag], anti-HBs Ag, anti-Toxoplasma gondii immunoglobulin G [IgG], anti-rubella virus IgG, anti-cytomegalovirus IgM, and anti-Epstein-Barr virus IgM) was evaluated for 375 samples. No difference was found among test results using fresh or stored frozen serum

  13. Comparison of miRNA quantitation by Nanostring in serum and plasma samples.

    Science.gov (United States)

    Foye, Catherine; Yan, Irene K; David, Waseem; Shukla, Neha; Habboush, Yacob; Chase, Lori; Ryland, Kristen; Kesari, Vivek; Patel, Tushar

    2017-01-01

    Circulating microRNAs that are associated with specific diseases have garnered much attention for use in diagnostic assays. However, detection of disease-associated miRNA can be affected by several factors such as release of contaminating cellular miRNA during sample collection, variations due to amplification of transcript for detection, or controls used for normalization for accurate quantitation. We analyzed circulating miRNA in serum and plasma samples obtained concurrently from 28 patients, using a Nanostring quantitative assay platform. Total RNA concentration ranged from 32-125 μg/ml from serum and 30-220 μg/ml from plasma. Of 798 miRNAs, 371 miRNAs were not detected in either serum or plasma samples. 427 were detected in either serum or plasma but not both, whereas 151 miRNA were detected in both serum and plasma samples. The diversity of miRNA detected was greater in plasma than in serum samples. In serum samples, the number of detected miRNA ranged from 3 to 82 with a median of 17, whereas in plasma samples, the number of miRNA detected ranged from 25 to 221 with a median of 91. Several miRNA such as miR451a, miR 16-5p, miR-223-3p, and mir25-3p were highly abundant and differentially expressed between serum and plasma. The detection of endogenous and exogenous control miRNAs varied in serum and plasma, with higher levels observed in plasma. Gene expression stability identified candidate invariant microRNA that were highly stable across all samples, and could be used for normalization. In conclusion, there are significant differences in both the number of miRNA detected and the amount of miRNA detected between serum and plasma. Normalization using miRNA with constant expression is essential to minimize the impact of technical variations. Given the challenges involved, ideal candidates for blood based biomarkers would be those that are indifferent to type of body fluid, are detectable and can be reliably quantitated.

  14. Comparison of miRNA quantitation by Nanostring in serum and plasma samples.

    Directory of Open Access Journals (Sweden)

    Catherine Foye

    Full Text Available Circulating microRNAs that are associated with specific diseases have garnered much attention for use in diagnostic assays. However, detection of disease-associated miRNA can be affected by several factors such as release of contaminating cellular miRNA during sample collection, variations due to amplification of transcript for detection, or controls used for normalization for accurate quantitation. We analyzed circulating miRNA in serum and plasma samples obtained concurrently from 28 patients, using a Nanostring quantitative assay platform. Total RNA concentration ranged from 32-125 μg/ml from serum and 30-220 μg/ml from plasma. Of 798 miRNAs, 371 miRNAs were not detected in either serum or plasma samples. 427 were detected in either serum or plasma but not both, whereas 151 miRNA were detected in both serum and plasma samples. The diversity of miRNA detected was greater in plasma than in serum samples. In serum samples, the number of detected miRNA ranged from 3 to 82 with a median of 17, whereas in plasma samples, the number of miRNA detected ranged from 25 to 221 with a median of 91. Several miRNA such as miR451a, miR 16-5p, miR-223-3p, and mir25-3p were highly abundant and differentially expressed between serum and plasma. The detection of endogenous and exogenous control miRNAs varied in serum and plasma, with higher levels observed in plasma. Gene expression stability identified candidate invariant microRNA that were highly stable across all samples, and could be used for normalization. In conclusion, there are significant differences in both the number of miRNA detected and the amount of miRNA detected between serum and plasma. Normalization using miRNA with constant expression is essential to minimize the impact of technical variations. Given the challenges involved, ideal candidates for blood based biomarkers would be those that are indifferent to type of body fluid, are detectable and can be reliably quantitated.

  15. Multitoxin analysis of Aspergillus clavatus-infected feed samples implicated in two outbreaks of neuromycotoxicosis in cattle in South Africa

    Directory of Open Access Journals (Sweden)

    Christo J. Botha

    2014-02-01

    Full Text Available Aspergillus clavatus intoxication is a highly fatal neuromycotoxicosis of ruminants, especially cattle. It is caused by the ingestion of infected sprouting grain and sorghum beer residue. Locomotor disturbances, tremors and paralysis are observed. Histologically, degeneration and necrosis of larger neurons in the medulla oblongata, the midbrain, the thalamus and the ventral horns of the spinal cord are observed. Although a range of mycotoxins such as patulin, cytochalasin E and pseurotin A have been isolated, there is limited information on which specific mycotoxin or group of mycotoxins are involved during outbreaks of intoxication in livestock. In the present study, two outbreaks of A. clavatus poisoning in cattle are briefly described. Feed samples were collected for fungal identification, and culture and multitoxin analysis. A range of fungal metabolites were detected, and the estimated concentrations (μg/kg are provided. Both the sprouting barley and brewer’s grain were predominantly infected with A. clavatus and, to a lesser extent, Rhizopus arrhizus. The only common Aspergillus secondary metabolite present in all the samples was pseurotin A. Patulin and cytochalasin E were present in the sprouting barley samples, as well as the A. clavatus isolates cultured on malt extract agar for 2 weeks; however, neither of these mycotoxins could be detected in the brewer’s grain sample.

  16. Genetic parameter estimates for serum insulin-like growth factor I concentrations, and body weight and weight gains in Angus beef cattle divergently selected for serum insulin-like growth factor I concentration.

    Science.gov (United States)

    Davis, M E; Simmen, R C M

    2006-09-01

    Data for the current study were obtained from a divergent selection experiment in which the selection criterion was the average serum IGF-I concentrations of 3 postweaning blood samples collected from purebred Angus calves. Multiple-trait derivative-free REML procedures were used to obtain genetic parameter estimates for IGF-I concentrations and for BW and BW gains measured from birth to the conclusion of a 140-d postweaning performance test. Included in the analysis were 2,674 animals in the A(-1) matrix, 1,761 of which had valid records for IGF-I concentrations. Direct heritability estimates +/- SE for IGF-I concentration at d 28, 42, and 56 of the postweaning period and for mean IGF-I concentrations were 0.44 +/- 0.07, 0.51 +/- 0.08, 0.42 +/- 0.07, and 0.52 +/- 0.08, respectively. Heritability estimates for maternal genetic effects ranged from 0.10 +/- 0.05 to 0.20 +/- 0.06. The proportion of total phenotypic variance due to the maternal permanent environmental effect was essentially zero for all measures of IGF-I concentrations. Genetic correlations of IGF-I concentrations with weaning and post-weaning BW ranged from 0.07 +/- 0.12 to 0.32 +/- 0.11 and generally demonstrated an increasing trend during the postweaning period. Averaged across the various measures of IGF-I, the genetic correlation of IGF-I with preweaning gain was 0.14, whereas the genetic correlation with postweaning gain was 0.29. Genetic correlations between IGF-I and BW gain were positive during all time intervals, except between weaning and the beginning of the postweaning test and from d 84 to 112 of the postweaning period. Environmental and phenotypic correlations of IGF-I with BW and BW gains were generally positive, but small. These results indicate that postweaning serum IGF-I concentration is moderately to highly heritable and has small positive genetic, environmental, and phenotypic correlations with BW other than birth weight and with pre- and postweaning gain. Therefore, if IGF

  17. Screening of different sample types associated with sheep and cattle for the presence of nematophagous fungi in China.

    Science.gov (United States)

    Cai, Kui-Zheng; Liu, Jun-Lin; Liu, Wei; Wang, Bo-Bo; Xu, Qiang; Sun, Long-Jie; Chen, Ming-Yue; Zhao, Ming-Wang; Wu, Jia-Yan; Li, Xiao-Shan; Yang, Jing; Wei, Shuan; Chen, Chun-Rong; Ma, Zhong-Ren; Xu, Chun-Lan; Wang, Feng; Hu, Qian-Lin; Fang, Wen-Xiu; Zheng, Tian-Hui; Wang, Yue-Ying; Zhu, Wen-Long; Li, Dan; Li, Qing; Zhang, Chao; Cai, Bing; Wang, Fan; Yang, Zai-Yun; Liu, Yan-Qiu

    2016-03-01

    A total of 1502 samples, including feces of sheep (793) and cattle (348), pasture soil (118), dung compost (147) and barn soil (96), were examined between October 2012 and August 2014 to discover potential strains of nematophagous fungi for the biological control of livestock-parasitic nematodes. These samples were collected from 87 sites located in 48 counties of 20 provinces (autonomous regions/municipalities) of China. Fungi were identified down to a species level. Four hundred and seventy-seven isolates, which were distributed in 8 genera and 28 taxa, were identified as nematophagous fungi. Nematode-trapping fungi included 17 species and one unidentified species of Arthrobotrys, two of Dactylella, Drechslerella dactyloides, and Duddingtonia flagrans. Five identified species and two unidentified species of endoparasitic fungi were isolated. The predominant species from all regions were Arthrobotrys oligospora, followed by Arthrobotrys musiformis, Arthrobotrys (Monacrosporium) thaumasiun, and Arthrobotrys (Monacrosporium) microscaphoides. Species with adhesive networks were the most frequently isolated. Among the endoparasitic fungi, Podocrella harposporifera (Harposporium anguillulae) was the most common species, followed by Harposporium lilliputanum and Harposporium arcuatum. Based on Shannon diversity index, the diversity levels of nematophagous fungi were relatively higher in samples associated with cattle, barn soil, and subtropical monsoon climate zone. Three species isolated from this study, namely, Duddingtonia flagrans, Arthrobotrys salina (Monacrosporium salinum), and Arthrobotrys oligospora var. sarmatica, are newly recorded in China, and 20 species (including one unidentified species) are newly recorded in sheep and cattle barn soils worldwide. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Evaluation of the illegal use of clenbuterol in Portuguese cattle farms from drinking water, urine, hair and feed samples.

    Science.gov (United States)

    Ramos, F; Baeta, M L; Reis, J; Silveira, M I N

    2009-06-01

    The recent discovery of clenbuterol contamination in Portuguese food led to the specific inspection of 16 cattle farms for beta-agonists, involving the analysis of a total of 486 samples (78 feed, 106 drinking water, 168 urine and 134 hair). The samples were screened for the beta-agonists: bromobuterol, cimaterol, clenbuterol, clenpenterol, clenproperol, hydroxymethylclenbuterol, mapenterol, salbutamol and terbutaline. Only clenbuterol was found in all analyzed matrices and the most likely method of illegal administration to animals was through drinking water. Of all samples analysed, 14.15% of drinking water were found positive in the range 0.03-3.80 mg l(-1) clenbuterol. Inclusion of hair samples in the Portuguese plan for clenbuterol residue control in live animals is discussed.

  19. Racial Differences in Serum Adipokine and Insulin Levels in a Matched Osteoarthritis Sample: A Pilot Study

    Directory of Open Access Journals (Sweden)

    Rajiv Gandhi

    2016-01-01

    Full Text Available Background. In an attempt to correlate biomarkers with disease, serum-based biomarkers often are compared between individuals with osteoarthritis (OA and control subjects. However, variable results have been reported. Some studies have suggested an association between certain adipokines and insulin and OA. We know that there are racial differences in OA prevalence and incidence, and from general population-based studies, those of Asian race consistently demonstrate a unique adipokine/insulin serum concentration profile as compared to Caucasians. Whether similar racial differences exist within OA samples is unknown and may have implications for selecting appropriate controls in comparative studies. Methods. Serum levels of adipokines, leptin, and adiponectin, along with insulin, were determined by ELISA in patients scheduled for total hip or knee replacement surgery for OA. Fifteen Asian patients were matched 1 : 1 on age (±2 years, gender, body mass index (±1.5 kg/m2, and surgical joint with Caucasian patients. Differences in serum concentrations were tested using paired t-tests. Results. Serum leptin and insulin levels were significantly higher in Asians compared to Caucasians (p<0.05. While serum adiponectin levels were lower among Asians, the difference did not reach statistical significance (p=0.12. Conclusion. Findings from this work suggest that when studying serum biomarker concentrations in OA versus controls, race may be an important factor to consider. Our findings warrant confirmation in larger studies.

  20. Differential Expression of Host Biomarkers in Saliva and Serum Samples from Individuals with Suspected Pulmonary Tuberculosis

    Directory of Open Access Journals (Sweden)

    Khutso G. Phalane

    2013-01-01

    Full Text Available The diagnosis of tuberculosis remains challenging in individuals with difficulty in providing good quality sputum samples such as children. Host biosignatures of inflammatory markers may be valuable in such cases, especially if they are based on more easily obtainable samples such as saliva. To explore the potential of saliva as an alternative sample in tuberculosis diagnostic/biomarker investigations, we evaluated the levels of 33 host markers in saliva samples from individuals presenting with pulmonary tuberculosis symptoms and compared them to those obtained in serum. Of the 38 individuals included in the study, tuberculosis disease was confirmed in 11 (28.9% by sputum culture. In both the tuberculosis cases and noncases, the levels of most markers were above the minimum detectable limit in both sample types, but there was no consistent pattern regarding the ratio of markers in serum/saliva. Fractalkine, IL-17, IL-6, IL-9, MIP-1β, CRP, VEGF, and IL-5 levels in saliva and IL-6, IL-2, SAP, and SAA levels in serum were significantly higher in tuberculosis patients (P<0.05. These preliminary data indicate that there are significant differences in the levels of host markers expressed in saliva in comparison to those expressed in serum and that inflammatory markers in both sample types are potential diagnostic candidates for tuberculosis disease.

  1. Rescue of foot-and-mouth disease viruses that are pathogenic for cattle from preserved viral RNA samples

    DEFF Research Database (Denmark)

    Belsham, Graham; Jamal, Syed Muhammad; Tjørnehøj, Kirsten

    2011-01-01

    of the rescued viruses (of serotype O and Asia 1) were inoculated into bull calves under high containment conditions. Acute clinical disease was observed in each case which spread rapidly from the inoculated calves to in-contact animals. Thus the rescued viruses were highly pathogenic. The availability......Background: Foot and mouth disease is an economically important disease of cloven-hoofed animals including cattle, sheep and pigs. It is caused by a picornavirus, foot-and-mouth disease virus (FMDV), which has a positive sense RNA genome which, when introduced into cells, can initiate virus...... replication. Principal Findings: A system has been developed to rescue infectious FMDV from RNA preparations generated from clinical samples obtained under experimental conditions and then applied to samples collected in the ‘‘field’’. Clinical samples from suspect cases of foot-and-mouth disease (FMD) were...

  2. The prevalence of serum antibodies to tick-borne infections in Mbale District, Uganda: The effect of agro-ecological zone, grazing management and age of cattle

    Directory of Open Access Journals (Sweden)

    C. Rubaire-Akiiki

    2004-03-01

    Full Text Available Between August and October 2000, a cross-sectional study was conducted in smallholder dairy farms in Mbale District, Uganda to assess the prevalence of ticks and tick-borne diseases under different grazing systems and agro-ecological zones and understand the circumstances under which farmers operated. A questionnaire was administered to obtain information on dairy farm circumstances and practices. A total of 102 farms were visited and sera and ticks were collected from 478 animals. Sero-prevalence of tick-borne diseases was determined using an enzyme-linked immunoassay. Acaricides were used indiscriminately but the intensity of their use varied with the grazing system and zone. Cattle from different farms mixed for various reasons. During the dry seasons farmers have to get additional fodder from outside their farms that can result in importation of ticks. The prevalence of ticks and serum antibodies to tick-borne infections differed across the grazing systems and zones. The highest serum antibody prevalence (>60% was recorded in the lowland zone under the free range and tethering grazing systems. The lowest tick challenge and serum antibody levels (<50% were recorded in the midland and upland zones under a zero-grazing system. These findings suggest that endemic stability to East Coast Fever, babesiosis and anaplasmosis is most likely to have existed in the lowland zone, particularly, under the tethering and free-range grazing systems. Also, endemic stability for babesiosis existed in the upland zones. Endemic instability for East Coast Fever existed in the midland and upland zones. These structured observational studies are instrumental in planning of control strategies for ticks and tick borne diseases since production systems and the cattle population at high risk of the diseases in the district have been identified.

  3. The prevalence of serum antibodies to tick-borne infections in Mbale District, Uganda: the effect of agro-ecological zone, grazing management and age of cattle.

    Science.gov (United States)

    Rubaire-Akiiki, C; Okello-Onen, J; Nasinyama, G W; Vaarst, M; Kabagambe, E K; Mwayi, W; Musunga, D; Wandukwa, W

    2004-01-01

    Between August and October 2000, a cross-sectional study was conducted in smallholder dairy farms in Mbale District, Uganda to assess the prevalence of ticks and tick-borne diseases under different grazing systems and agro-ecological zones and understand the circumstances under which farmers operated. A questionnaire was administered to obtain information on dairy farm circumstances and practices. A total of 102 farms were visited and sera and ticks were collected from 478 animals. Sero-prevalence of tick-borne diseases was determined using an enzyme-linked immunoassay. Acaricides were used indiscriminately but the intensity of their use varied with the grazing system and zone. Cattle from different farms mixed for various reasons. During the dry seasons farmers have to get additional fodder from outside their farms that can result in importation of ticks. The prevalence of ticks and serum antibodies to tick-borne infections differed across the grazing systems and zones. The highest serum antibody prevalence (>60%) was recorded in the lowland zone under the free range and tethering grazing systems. The lowest tick challenge and serum antibody levels (<50%) were recorded in the midland and upland zones under a zero-grazing system. These findings suggest that endemic stability to East Coast Fever, babesiosis and anaplasmosis is most likely to have existed in the lowland zone, particularly, under the tethering and free-range grazing systems. Also, endemic stability for babesiosis existed in the upland zones. Endemic instability for East Coast Fever existed in the midland and upland zones. These structured observational studies are instrumental in planning of control strategies for ticks and tick borne diseases since production systems and the cattle population at high risk of the diseases in the district have been identified.

  4. Per- and polyfluoroalkyl substances in human serum and urine samples from a residentially exposed community.

    Science.gov (United States)

    Worley, Rachel Rogers; Moore, Susan McAfee; Tierney, Bruce C; Ye, Xiaoyun; Calafat, Antonia M; Campbell, Sean; Woudneh, Million B; Fisher, Jeffrey

    2017-09-01

    Per- and polyfluoroalkyl substances (PFAS) are considered chemicals of emerging concern, in part due to their environmental and biological persistence and the potential for widespread human exposure. In 2007, a PFAS manufacturer near Decatur, Alabama notified the United States Environmental Protection Agency (EPA) it had discharged PFAS into a wastewater treatment plant, resulting in environmental contamination and potential exposures to the local community. To characterize PFAS exposure over time, the Agency for Toxic Substances and Disease Registry (ATSDR) collected blood and urine samples from local residents. Eight PFAS were measured in serum in 2010 (n=153). Eleven PFAS were measured in serum, and five PFAS were measured in urine (n=45) from some of the same residents in 2016. Serum concentrations were compared to nationally representative data and change in serum concentration over time was evaluated. Biological half-lives were estimated for perfluorooctanoic acid (PFOA), perfluorooctane sulfonic acid (PFOS), and perfluorohexane sulfonic acid (PFHxS) using a one-compartment pharmacokinetic model. In 2010 and 2016, geometric mean PFOA and PFOS serum concentrations were elevated in participants compared to the general U.S. In 2016, the geometric mean PFHxS serum concentration was elevated compared to the general U.S. Geometric mean serum concentrations of PFOA, PFOS, and perfluorononanoic acid (PFNA) were significantly (p≤0.0001) lower (49%, 53%, and 58%, respectively) in 2016 compared to 2010. Half-lives for PFOA, PFOS, and PFHxS were estimated to be 3.9, 3.3, and 15.5years, respectively. Concentrations of PFOA in serum and urine were highly correlated (r=0.75) in males. Serum concentrations of some PFAS are decreasing in this residentially exposed community, but remain elevated compared to the U.S. general population. Published by Elsevier Ltd.

  5. Production of transgenic cattle highly expressing human serum albumin in milk by phiC31 integrase-mediated gene delivery.

    Science.gov (United States)

    Luo, Yan; Wang, Yongsheng; Liu, Jun; Lan, Hui; Shao, Minghao; Yu, Yuan; Quan, Fusheng; Zhang, Yong

    2015-10-01

    Transgenic cattle expressing high levels of recombinant human serum albumin (HSA) in their milk may as an alternative source for commercial production. Our objective was to produce transgenic cattle highly expressing HSA in milk by using phiC31 integrase system and somatic cell nuclear transfer (SCNT). The mammary-specific expression plasmid pIACH(-), containing the attB recognition site for phiC31 integrase, were co-transfected with integrase expression plasmid pCMVInt into bovine fetal fibroblast cells (BFFs). PhiC31 integrase-mediated integrations in genome of BFFs were screened by nested inverse PCR. After analysis of sequence of the PCR products, 46.0% (23/50) of the both attB-genome junction sites (attL and attR) were confirmed, and four pseudo attP sites were identified. The integration rates in BF3, BF11, BF19 and BF4 sites were 4.0% (2/50), 6.0% (3/50), 16.0% (8/50) and 20.0% (10/50), respectively. BF3 is located in the bovine chromosome 3 collagen alpha-3 (VI) chain isomer 2 gene, while the other three sites are located in the non-coding region. The transgenic cell lines from BF11, BF19 and BF4 sites were used as donors for SCNT. Two calves from transgenic cells BF19 were born, one died within a few hours after birth, and another calf survived healthy. PCR and Southern blot analysis revealed integration of the transgene in the genome of cloned calves. The nested reverse PCR confirmed that the integration site in cloned calves was identical to the donor cells. The western blotting assessment indicated that recombinant HSA was expressed in the milk of transgenic cattle and the expression level was about 4-8 mg/mL. The present study demonstrated that phiC31 integrase system was an efficient and safety gene delivery tool for producing HSA transgenic cattle. The production of recombinant HSA in the milk of cattle may provide a large-scale and cost-effective resource.

  6. Use of an immunoaffinity column for tetrachlorodibenzo-p-dioxin serum sample cleanup.

    Science.gov (United States)

    Shelver, W L; Larsen, G L; Huwe, J K

    1998-02-13

    Covalently linking 1-amino-3,7,8-trichlorodibenzo-p-dioxin with either keyhole limpet hemocyanin (KLH) or bovine serum albumin (BSA) provided antigens that generated antibodies in chickens. Competitive ELISA analysis demonstrated that the antibodies isolated from egg yolk (IgY) bound with 1,3,7,8-tetrachlorodibenzo-p-dioxin (1,3,7,8-TCDD). The antibodies were linked to CNBr-Sepharose to generate an immunoaffinity column. Radiolabeled 1,3,7,8-TCDD in a 0.05% Tween 20 solution was retained by the column and could be eluted by increasing the Tween 20 concentration. The binding efficiency for 10.7 ng per ml gel matrix ranged from 85 to 97%. Immunoaffinity columns generated by this method did not effectively bind 14C-1,3,7,8-TCDD from serum samples. Diluting the serum 1:20 with 0.05% Tween 20 increased the binding efficiency. Alternately, ethanol-hexane extraction followed by solid phase extraction on a carbon column using a fat removal protocol also provided an appropriate preaffinity column cleanup for serum samples. After this preaffinity column cleanup, spiked serum samples applied to the immunoaffinity column showed binding efficiencies of over 90%.

  7. Serial Sampling of Serum Protein Biomarkers for Monitoring Human Traumatic Brain Injury Dynamics: A Systematic Review

    Directory of Open Access Journals (Sweden)

    Eric Peter Thelin

    2017-07-01

    Full Text Available BackgroundThe proteins S100B, neuron-specific enolase (NSE, glial fibrillary acidic protein (GFAP, ubiquitin carboxy-terminal hydrolase L1 (UCH-L1, and neurofilament light (NF-L have been serially sampled in serum of patients suffering from traumatic brain injury (TBI in order to assess injury severity and tissue fate. We review the current literature of serum level dynamics of these proteins following TBI and used the term “effective half-life” (t1/2 in order to describe the “fall” rate in serum.Materials and methodsThrough searches on EMBASE, Medline, and Scopus, we looked for articles where these proteins had been serially sampled in serum in human TBI. We excluded animal studies, studies with only one presented sample and studies without neuroradiological examinations.ResultsFollowing screening (10,389 papers, n = 122 papers were included. The proteins S100B (n = 66 and NSE (n = 27 were the two most frequent biomarkers that were serially sampled. For S100B in severe TBI, a majority of studies indicate a t1/2 of about 24 h, even if very early sampling in these patients reveals rapid decreases (1–2 h though possibly of non-cerebral origin. In contrast, the t1/2 for NSE is comparably longer, ranging from 48 to 72 h in severe TBI cases. The protein GFAP (n = 18 appears to have t1/2 of about 24–48 h in severe TBI. The protein UCH-L1 (n = 9 presents a t1/2 around 7 h in mild TBI and about 10 h in severe. Frequent sampling of these proteins revealed different trajectories with persisting high serum levels, or secondary peaks, in patients with unfavorable outcome or in patients developing secondary detrimental events. Finally, NF-L (n = 2 only increased in the few studies available, suggesting a serum availability of >10 days. To date, automated assays are available for S100B and NSE making them faster and more practical to use.ConclusionSerial sampling of brain-specific proteins in serum reveals

  8. Benefits and Limits of Egg Yolk vs. Serum Samples for Avian Influenza Virus Serosurveillance.

    Science.gov (United States)

    Abdelwhab, E M; Grund, Christian; Aly, Mona M; Beer, Martin; Harder, Timm C; Hafez, Hafez M

    2016-06-01

    Serologic tests are a valuable tool for retrospective surveillance of avian influenza viruses (AIV) and monitoring of postvaccination host immune response. Yet collection of serum samples, particularly in adult breeder chickens, is laborious, intrusive to birds, and may pose a serious risk to the biosecurity of a flock. In this study we compared the level of AIV-specific antibody titers in eggs and serum samples obtained from broiler breeder chickens vaccinated at 6, 12, and 18 wk of age with H5N2-inactivated vaccine. Nucleocapsid protein-specific ELISA and hemagglutination inhibition test (HI) against homologous as well as heterologous antigens were used. The eggs and sera were collected at 22, 30, 45, and 50 wk of age (i.e., 4, 12, 27, and 32 wk after the third and final immunization, respectively). Using ELISA, the number of positive egg yolk samples decreased over time after vaccination, from 97% to 47%, while the seropositivity rate of serum samples was 97%-100% during the whole investigation period. No antibody titers were detected in egg white. By HI, antibody titers in serum samples were higher than in egg yolk samples. Compared to the homologous H5N2 antigen, significantly lower HI titers were obtained by using a heterologous H5N1 virus of clade 2.2.1.2. In addition, no HI titers were detected in egg yolk and/or serum samples tested against the antigen of an Egyptian H5N1 antigenic drift variant of clade 2.2.1.1. This study indicates that egg yolk may be used to monitor the postvaccination immune status of broiler breeder chickens and retrospective serosurveillance-by HI when a matching antigen is available as well as by ELISA-particularly for up to 12 wk postvaccination.

  9. A flow injection sampling resonance light scattering system for total protein determination in human serum

    Science.gov (United States)

    Dong, Lijun; Li, Ying; Zhang, Yaheng; Chen, Xingguo; Hu, Zhide

    2007-04-01

    A novel flow injection method with resonance light scattering detection was developed for the determination of total protein concentrations. This method is based on the enhancement of RLS signals from Methyl Blue (MB) by protein. The enhanced RLS intensities at 333 nm, in a pH 4.1 acidic aqueous solution, were proportional to the protein concentration over the range 2.0-37.3 and 1.0-36.0 μg ml -1 for human serum albumin (HSA) and bovine serum albumin (BSA), respectively. The corresponding limits of detection (3 σ) of 45 ng ml -1 for HSA and 80 ng ml -1 for BSA were attained. The method was successfully applied to the quantification of total proteins in human serum samples, the maximum relative error is less than 1% and the recovery is between 98% and 102%. The sample throughput was 60 h -1.

  10. Significant Decline in Galactomannan Signal during Storage of Clinical Serum Samples

    Directory of Open Access Journals (Sweden)

    Samir G. Agrawal

    2013-06-01

    Full Text Available Galactomannan (GM is widely used for detection of invasive aspergillosis in high-risk haemato-oncology patients. Recent publications have reported a lack of repeatability of GM detection. The objective of this retrospective study was to assess the repeatability of GM levels during storage of clinical samples. In a GM screening strategy, positive sera were repeat tested as per manufacturer’s recommendations. Short-term (ST storage of samples was at +4 °C while long-term (LT storage was at −80 °C. Bronchoalveolar (BAL fluid was also repeating tested after ST storage and LT storage. Wilcoxon Signed Ranks Test was employed to assess the repeatability of GM levels. In a subset of 14 GM positive sera, repeat testing was performed on both the original serum and ethylenediaminetetraacetic acid (EDTA pre-treated sample. There was a significant reduction in GM signals on repeat testing following ST storage (median GM index: 0.65 vs. 0.19; p < 0.001 and LT storage (median GM index: 0.56 vs. 0.10; p < 0.001 of serum samples. Of samples that were initially GM positive, an average GM index reduction of 50% was seen, with approximately two-thirds becoming GM negative on repeat testing of the same sample. In contrast, GM signal loss was not seen on repeat testing of BAL fluid following ST or LT storage. When GM positive serum samples were repeat tested using EDTA pre-treated serum from the first step of the testing protocol, all samples remained GM positive. In contrast, when the same samples were repeat tested from the original collected serum, 9 samples (64% became GM negative. The significant reduction in GM signals during ST and LT storage of serum samples has implications for clinical management. Although the reasons for GM decline are unknown, they occur prior to the EDTA pre-treatment stage, indicating that the time from phlebotomy to testing should be minimized. BAL fluid GM index values remain stable.

  11. Predictors of Serum 25-Hydroxyvitamin D Concentrations among a Sample of Egyptian Schoolchildren

    Directory of Open Access Journals (Sweden)

    Mones M. Abu Shady

    2016-01-01

    Full Text Available Objective. To assess the level of 25-hydroxyvitamin D status among a sample of Egyptian schoolchildren and to evaluate predictors of deficiency and insufficiency. Subjects and Methods. A cross-sectional study comprising 200 prepubescent schoolchildren aged from 9 to 11 years was performed. A questionnaire including frequency of midday sun exposure, milk intake, physical activity, and level of maternal education was taken. Body mass index (BMI was calculated; serum 25-hydroxyvitamin D [25(OHD], serum calcium, phosphorus, and parathyroid hormone were measured. Results. Vitamin D deficiency [serum 25(OHD < 20 ng/mL] was detected in 11.5% of subjects while its insufficiency (serum 25(OHD is between 20 and 29.9 ng/mL was detected in 15%. Results revealed that obesity, low physical activity, low sun exposure, and low maternal education level are significant predictors of insufficiency, though female gender, low maternal education level, and low milk intake are significant predictors of deficiency. Lower serum phosphorus and higher serum parathyroid hormone were significantly associated with both deficiency and insufficiency (p<0.05. Conclusion. Vitamin D deficiency and insufficiency are common among schoolchildren in Egypt. Food fortification, vitamin D supplementation, and increasing maternal awareness about the importance of physical activity and exposure of their children to ultraviolet light may help to overcome this problem.

  12. Changes in alt, ast and alp values of plasma and serum samples ...

    African Journals Online (AJOL)

    This study investigated the time-related changes in the alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) values of serum and plasma samples stored at refrigerator (2 - 8oC, average of 40C) and room temperature (30-340C, average of 320C) for a period of 120 hours (5 ...

  13. Differential expression of small non-coding RNAs in serum from cattle challenged with viruses causing bovine respiratory disease

    Science.gov (United States)

    MicroRNAs and tRNA-derived RNA fragments (tRFs) are the two most abundant groups of small non-coding RNAs. The potential for microRNAs and tRFs to be used as pathogen exposure indicators is yet to be fully explored. Our objective was to identify microRNAs and tRFs in cattle challenged with a non-cy...

  14. Modulation of Cytokine mRNA Expression in Pharyngeal Epithelial Samples obtained from Cattle Infected with Foot-and-Mouth Disease Virus

    DEFF Research Database (Denmark)

    Stenfeldt, Anna Carolina; Heegaard, Peter M. H.; Stockmarr, Anders

    2012-01-01

    A novel technique of endoscopical collection of small tissue samples was used to obtain sequential tissue samples from the dorsal soft palate (DSP) of individual cattle infected with foot-and-mouth disease virus (FMDV) at different phases of the infection. Levels of mRNA encoding interferon (IFN...

  15. Gas chromatographic/mass spectrometric determination of lysergic acid diethylamide (LSD) in serum samples.

    Science.gov (United States)

    Musshoff, F; Daldrup, T

    1997-08-04

    A sensitive method for the detection and quantification of lysergic acid diethylamide (LSD) in serum samples is described. After liquid-liquid extraction the trimethylsilyl derivative of LSD is detected by gas chromatography-mass spectrometry. Experiments with spiked samples resulted in a recovery of 76%, the coefficient of variation was 9.3%. Excellent linearity was obtained over the range 0.1-10 ng ml-1. Additionally experiments demonstrating the light sensitivity of LSD are presented together with casuistics.

  16. Seroprevalence of Neospora caninum Infection in Dairy Cattle in Tabriz, Northwest Iran

    Science.gov (United States)

    Nematollahi, A; Jaafari, R; Moghaddam, Gh

    2011-01-01

    Background The aim of this study was to determine the seroprevalence of antibody to Neospora caninum in healthy and aborted dairy cattle in Tabriz, capital of East-Azarbaijan in northwest of Iran. Methods In this cross-sectional study serum samples were collected from 266 healthy and aborted Holestein-Feriesisnc cows from September 2008 to August 2009. The sera were analyzed to detect of antibody against N. caninum using the commercially ELISA kit. Results Seroprevalence of antibody to N. caninum was 10.5% in Tabriz dairy cattle. Also the abortion rate in all cattle sampled was 33.6% but percentage of seropositive aborted cattle was 18.4%. Conclusion Neosporosis could be one of the possible causes of abortion in dairy cattle in Tabriz and regarding the distribution in dogs as definitive host for the parasite, further studies in dog and cattle are recommended. PMID:22347319

  17. Preanalytical variables affecting the quantification of fatty acid ethyl esters in plasma and serum samples.

    Science.gov (United States)

    Soderberg, B L; Sicinska, E T; Blodget, E; Cluette-Brown, J E; Suter, P M; Schuppisser, T; Vetter, W; Laposata, M

    1999-12-01

    Fatty acid ethyl esters (FAEEs) are cytotoxic nonoxidative ethanol metabolites produced by esterification of fatty acids and ethanol. FAEEs are detectable in blood up to 24 h after ethanol consumption. The objective of this study was to assess the impact of gender, serum or plasma triglyceride concentration, time and temperature of specimen storage, type of alcoholic beverage ingested, and the rate of ethanol consumption on FAEE concentrations in plasma or serum. For some studies, subject were recruited volunteers; in others, residual blood samples after ethanol quantification were used. FAEEs were isolated by solid-phase extraction and quantified by gas chromatography-mass spectrometry. For weight-adjusted amounts of ethanol intake, FAEE concentrations were twofold greater for men than women (P /=24 h. The type of alcoholic beverage and rate of consumption did not affect FAEE concentrations. These studies advance plasma and serum FAEE measurements closer to implementation as a clinical test for ethanol intake.

  18. Rapid endoglin determination in serum samples using an amperometric magneto-actuated disposable immunosensing platform.

    Science.gov (United States)

    Torrente-Rodríguez, Rebeca M; Campuzano, Susana; Ruiz-Valdepeñas-Montiel, Víctor; Pedrero, María; Fernández-Aceñero, M Jesús; Barderas, Rodrigo; Pingarrón, José M

    2016-09-10

    A sensitive and rapid method for the determination of the clinically relevant biomarker human endoglin (CD105) in serum samples is presented, involving a magneto-actuated immunoassay and amperometric detection at disposable screen-printed carbon electrodes (SPCEs). Micro-sized magnetic particles were modified with a specific antibody to selectively capture the target protein which was further sandwiched with a secondary HRP-labeled antibody. The immunocomplexes attached to the magnetic carriers were amperometrically detected at SPCEs using the hydroquinone (HQ)/H2O2/HRP system. The magneto-actuated immunosensing platform was able to detect 5 pmoles of endoglin (in 25μL of sample, 0.2μM) in 30min providing statistically similar results to those obtained using a commercial ELISA kit for the determination of endogenous content of endoglin in human serum samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. The Effect of Sample Storage Temperature on the Phenytoin Level in Serum

    Directory of Open Access Journals (Sweden)

    Paiboon Tummarintra

    2015-11-01

    Full Text Available Objective: To study the effect of temperature on phenytoin concentration in serum and to determine the sample were kept at 2-8°C and 35°C. Methods: Serum samples of patients, who had been treated with phenytoin. had their phenytoin level determined on the first day. The sample was stored for seven days and analyzed on days three, five and seven of the study and divided into three storage concentrations from 2.5-10, 10.1-20 and 20.1-30 μg/ml compared using 50 samples per group. Results: Phenytoin level was found to have increased over time to three days at a temperature of 35°C and five days at 2-8°C. It increased in the level of statistical significance at p<0.05. This study’s data were all well correlated at r2 = 0.9981-0.9999. Conclusion: Collected samples for analysis of phenytoin level must be stored at 2-8°C and stored for at least five days, to prevent degradation of protein which cannot bind to the drug. The phenytoin levels in serum increased to make the diagnosis of dosage mistakes.

  20. Serum Chrome levels sampled with steel needle vs. plastic IV cannula

    DEFF Research Database (Denmark)

    Penny, Jeannette Østergaard; Overgaard, Søren

    2010-01-01

    . This study aimed to test that theory. Method: We compared serum chromium values for two sampling methods, steel needle and IV plastic cannula, as well as sampling sequence in 16 healthy volunteers. Results: We found statistically significant chromium contamination from the steel needle with mean differences...... between the two methods of 0.073 ng/mL, for the first sample, and 0.033 ng/mL for the second. No difference was found between the first and second plastic sample. The first steel needle sample contained an average of 0.047 ng/mL more than the second. This difference was only borderline significant....... Conclusion: The chromium contamination from the steel needle is low, and sampling method matters little in MoM populations. If using steel needles we suggest discarding the first sample....

  1. Diversity of spore-forming bacteria in cattle manure, slaughterhouse waste and samples from biogas plants

    National Research Council Canada - National Science Library

    Bagge, E; Persson, M; Johansson, K-E

    2010-01-01

    .... In this study, the effect of the biogas process on Bacillus spp. and Clostridium spp. was investigated. We analysed 97 faecal samples, 20 slaughterhouse waste samples and 60 samples collected at different stages in the biogas process...

  2. Evaluation of Sample Handling Effects on Serum Vitamin E and Cholesterol Concentrations in Alpacas

    Directory of Open Access Journals (Sweden)

    Andrea S. Lear

    2014-01-01

    Full Text Available Clinical cases of vitamin E deficiencies have been diagnosed in camelids and may indicate that these species are more sensitive to inadequate vitamin E in hay-based diets compared to other ruminant and equine species. In bovine, cholesterol has been reported to affect vitamin E concentrations. In order to evaluate vitamin E deficiencies in camelids, the effects of collection and storage of the blood samples prior to processing were necessary. Reports vary as to factors affecting vitamin E and cholesterol in blood samples, and diagnostic laboratories vary in instructions regarding sample handling. Blood was collected from healthy alpacas and processed under conditions including exposure to fluorescent light, serum and red blood cell contact, tube stopper contact, temperature, and hemolysis. Serum vitamin E and cholesterol concentrations were then measured. Statistical analyses found that the vitamin E concentrations decreased with prolonged contact with the tube stopper and with increasing hemolysis. Vitamin E concentration variations were seen with other factors but were not significant. Time prior to serum separation and individual animal variation was found to alter cholesterol concentrations within the sample, yet this finding was clinically unremarkable. No correlation was seen between vitamin E and cholesterol concentration, possibly due to lack of variation of cholesterol.

  3. IgM-capture ELISA of serum samples collected from Filipino dengue patients.

    Science.gov (United States)

    Buerano, C C; Ibrahim, I N; Contreras, R C; Hasebe, F; Matias, R R; Natividad, F F; Igarashi, A

    2000-09-01

    Viral antigens for 4 dengue serotypes were produced in C6/36 Aedes albopictus cells. These were used as assay antigens for IgM-capture ELISA to detect IgM antibodies in sera of dengue patients from 3 hospitals in Metro Manila, Philippines. A total of 378 serum samples came from National Children's Hospital (NCH), San Lazaro Hospital (SLH), and St Luke's Medical Center (SLMC), from January to November 1995. Three hundred and four (304) out of 378 serum samples, or 80.42% showed positive IgM ELISA titer against at least one of the 4 assay antigens. Dengue type 4 (D4) antigen detected antibodies in 61.90% (234/378) of these serum samples, whereas type 1 (D1), type 3 (D3), and type 2 (D2) had detection rates of 60.05% (227/378), 50.79% (192/378) and 49.47% (187/378) respectively. Although the results show that both D1 and D4 are the most effective antigens in identifying dengue infections for this batch of samples, the use of a cocktail of antigens is still recommended. The results of this study are the basis for the IgM-capture ELISA protocol presently applied for the laboratory confirmation of dengue cases in the Philippines.

  4. Application of PCR in serum samples for diagnosis of paracoccidioidomycosis in the southern Bahia-Brazil.

    Science.gov (United States)

    Dias, Lucas; de Carvalho, Leila Falcão; Romano, Carla C

    2012-01-01

    Paracoccidioidomycosis (PCM) cannot always be diagnosed by conventional means such as direct examination of histopathology or clinical samples, and serological methods, used as an alternative, still have many cases of cross-reactivity. In this scenario, molecular techniques seem to arise as a rapid approach, specific and direct that could be used in the diagnosis of this mycosis. In this study we analyzed 76 serum samples from patients in southern Bahia suspected of having paracoccidioidomycosis using a conventional PCR with primers for the ITS1 ribosomal DNA of P. brasiliensis. Of these 76 patients, 5 were positive for PCM by double immunodiffusion and/or direct examination and histopathology. To test specificity of PCR, we used human DNA and three isolates of P. lutzii (1578, 01 and ED01). Additionally, we analyzed by serial dilutions of DNA the limit of detection of the assay. The test of PCR proved specific, as only a 144 bp fragment of the three isolates of P. lutzii and no human DNA was amplified. Detection limit was 1.1 pg/µL of DNA. Despite the high detection limit and specificity of PCR none of the 76 serum samples were found positive by PCR, but a biopsy specimen obtained from one of the patients with PCM was positive. These results, albeit limited, show that PCR is not effective in detecting DNA of P. brasiliensis or P. lutzii in serum, but could perhaps be used with other types of clinical samples, especially in those instances in which conventional methods fail.

  5. Mineral contents of extracellular fluids in camel and cattle in the ...

    African Journals Online (AJOL)

    Serum, urine, and rumen samples were obtained from the animals and analysed for calcium, phosphorus, magnesium, sodium, potassium, and copper. The results showed that the concentrations of all the minerals studied, except sodium, were higher in the ruminal fluid of camels compared to cattle. However, serum ...

  6. Serum chromium levels sampled with steel needle versus plastic IV cannula. Does method matter?

    DEFF Research Database (Denmark)

    Penny, Jeannette Ø; Overgaard, Søren

    2010-01-01

    PURPOSE: Modern metal-on-metal (MoM) joint articulations releases metal ions to the body. Research tries to establish how much this elevates metal ion levels and whether it causes adverse effects. The steel needle that samples the blood may introduce additional chromium to the sample thereby...... causing bias. This study aimed to test that theory. METHODS: We compared serum chromium values for two sampling methods, steel needle and IV plastic cannula, as well as sampling sequence in 16 healthy volunteers. RESULTS: We found statistically significant chromium contamination from the steel needle...... significant. CONCLUSION: The chromium contamination from the steel needle is low, and sampling method matters little in MoM populations. If using steel needles we suggest discarding the first sample....

  7. Prevalence of bacteremia in dairy cattle with acute puerperal metritis.

    Science.gov (United States)

    Credille, B C; Woolums, A R; Giguère, S; Robertson, T; Overton, M W; Hurley, D J

    2014-01-01

    Acute puerperal metritis (APM) affects 30% of postpartum dairy cattle. Bacteremia negatively impacts survival in cattle with coliform mastitis. However, the prevalence of bacteremia in dairy cattle with APM is unknown. Bacteremia is detectable in a large proportion of cattle with APM. Seventeen dairy cows with APM and 17 healthy dairy cattle. Prospective case-control study. Cases were identified by daily monitoring of cattle in the first 10 days after calving. Controls were matched to cases by parity and days in milk. Cows were examined at the time of identification of APM. A complete blood count, serum biochemical analysis, and bacteriologic culture of blood and lochial fluid were performed on each animal at the time of diagnosis. The same samples were collected from healthy herdmates of a similar parity and days in milk. Blood culture results and clinicopathologic variables were compared between groups. Conditional logistic regression was used to evaluate factors associated with APM, whereas multivariate logistic regression was used to evaluate factors associated with bacteremia. Bacteremia occurred in 53% (9/17) of cattle with APM and 53% (8/15) controls. Bacillus spp. was the organism most commonly isolated from the bloodstream in cattle of both groups. Bacteremic cattle in both groups were significantly less likely to have basophils in the peripheral circulation (P = .02) and more likely to have higher serum globulin concentrations (P = .02). Bacteremia is a common occurrence in postpartum dairy cattle. Further study is warranted to investigate the modes by which bacteria colonize the bloodstream in this population of animals and the importance of bacteremia on health and productivity of affected animals. Copyright © 2014 by the American College of Veterinary Internal Medicine.

  8. Quality assesment for the analysis of PCDDs/PCDFs in individual human serum samples

    Energy Technology Data Exchange (ETDEWEB)

    Perez, F. [IIQAB-CSIC, Barcelona (Spain). Dept. of Ecotechnologies, Lab. of Dioxins; Abad, E.; Llerena, J.J.; Caixach, J.; Rivera, J.

    2004-09-15

    The aim of this work was to optimise a relevant methodology for the ultratrace analysis of PCDDs/PCDFs in individual human serum samples. In order to carry out the study, different strategies including the elaboration of quality control samples, parallel sample analysis, control blanks and a number of quality assurance measures were implemented as analytical current practices. Some of the main drawbacks in the analysis of PCDDs/PCDFs in these kind of samples come from two conflicting aspects: the small sample size and the low levels expected to be found. Taking this into account, an unavoidable compromise between the sample amount and the minimum analytical requirements, mainly the detection limit (LOD), is mandatory. To reach this goal C{sub 18} solid phase extraction was used to remove the analytes from the matrix. Clean up was performed by solid-liquid adsorption chromatography using a variety of adsorbents. Instrumental analysis was achieved by high-resolution gas chromatography coupled to high-resolution mass spectrometry (HRGC/HRMS). Finally, the optimised methodology was applied to evaluate the potential impact in general population living in the surroundings of an obsolete municipal waste incinerator plant (MWI). Thus, more than 400 individuals serum samples potentially exposed to the emission of the incinerator and people not exposed were considered in this study.

  9. Detection of antibodies to Salmonella lipopolysaccharide in muscle fluid from cattle

    DEFF Research Database (Denmark)

    Hoorfar, Jeffrey; Wedderkopp, A.; Lind, Peter

    1997-01-01

    -enzootic areas of Jutland (n = 144). Procedure-Salmonella dublin (O:1,9,12), S typhimurium (O:1,4,5,12), and Salmonella O:9-blocking ELISA were used for testing the samples. Results-In the S dublin ELISA, all serum and muscle fluid samples from cattle on the island of Bornholm had OD450 values well below......Objective-To compare muscle fluid with serum samples for detection of antibodies to Salmonella lipopolysaccharide. Sample Population-Muscle fluid and serum samples from 2 cattle populations: 1 from the island of Bornholm with no history of salmonellosis (n = 39), and the other from the S dublin......%) and the O:9-blocking ELISA (r(s) = 0.49, P Salmonella infections in cattle....

  10. Evaluation of data loggers, sampling intervals, and editing techniques for measuring the lying behavior of dairy cattle.

    Science.gov (United States)

    Ledgerwood, D N; Winckler, C; Tucker, C B

    2010-11-01

    Lying behavior in dairy cattle can provide insight into how cows interact with their environment. Although lying behavior is a useful indicator of cow comfort, it can be time consuming to measure. In response to these time constraints, using data loggers to automate behavioral recording has become increasingly common. We tested the accuracy of the Onset Pendant G data logger (Onset Computer Corporation, Bourne, MA) for measuring lying behavior in dairy cattle (n=24 cows; 12 in each of 2 experiments). Cows wore the logger on the lateral (experiment 1) or medial (experiment 2) side of the hind leg above the metatarsophalangeal joint. Loggers recorded behavior at 4 sampling intervals (6, 30, 60, and 300 s) for at least 1.5 d. Data were smoothed using 3 editing methods to examine the effects of short, potentially erroneous readings. For this purpose, Microsoft Excel macros (Microsoft Corp., Redmond, WA) converted readings (i.e., lying events bordered by standing or vice versa) occurring singly or in consecutive runs of ≤2 or ≤6. Behavior was simultaneously recorded with digital video equipment. The logger accurately measured lying and standing. For example, predictability, sensitivity, and specificity were >99% using 30-s sampling and the single-event filter compared with continuously scored video recordings. The 6- and 30-s sampling intervals were comparable for all aspects of lying behavior when short events were filtered from the data set. Estimates of lying time generated from the 300-s interval unfiltered regimen were positively related (R(2) ≥ 0.99) to estimates of lying time from video, but this sampling regimen overestimated the number of lying bouts. This is likely because short standing and lying bouts were missed (12 and 34% of lying and standing bouts were <300 s in experiment 1 and 2, respectively). In summary, the data logger accurately measured all aspects of lying behavior when the sampling interval was ≤30 s and when short readings of lying and

  11. Use of real-time PCR on faecal samples for detection of sub-clinical Salmonella infection in cattle did not improve the detection sensitivity compared to conventional bacteriology

    DEFF Research Database (Denmark)

    Jensen, Annette Nygaard; Nielsen, L.R.; Baggesen, Dorte Lau

    2013-01-01

    bacteriological culture-reference method (BCRM) on cattle faecal samples for detection of sub-clinical Salmonella infections in cattle. Thirty faecal samples were artificially contaminated with either 10 or 50CFU of one of five strains of S. Dublin (SD) and S. Typhimurium (ST). The overall detection sensitivity...

  12. The Distribution Of Bovine Leukemia Virus Genotypes In Cattle From ...

    African Journals Online (AJOL)

    A study to investigate the types and distribution of bovine leukemia virus (BLV) genotypes and to estimate diagnostic test performance of agar gel immunodiffusion (AGID) was conducted on 807 cattle drawn from 68 farms found in 16 prefectures in Japan from. June 2002 to December 2003. AGID test on serum samples and ...

  13. THE INFLUENCE OF SINGLE NUCLEOTIDE POLYMORPHISM IN THE LEPTIN GENE ON SERUM ENERGY METABOLISM PARAMETERS IN CATTLE

    Directory of Open Access Journals (Sweden)

    Lenka Kovarikova

    2015-02-01

    Full Text Available The aim of this study was to test hypothesis that genotype TT in Czech Pied bulls, associated with higher leptinemia, will have significant effect on energy metabolism parameters in blood serum. The experiment was performed on the Czech Pied bulls in average age of 253 ± 13 days. The animals were sorted out into three groups according to the leptin genotype. The genotype TT was represented 20 animals, CT was represented 143 animals and CC was represented 127 animals. In the blood serum β-hydroxybutyrate (BHB and non-esterified fatty acid (NEFA were analyzed. One-way ANOVA revealed significant relation between serum concentration of NEFA and SNP genotype in leptin gene of Czech Pied bulls, but we failed to verify the hypothesis that genotype TT could affect some of monitored parameters.

  14. Fast Determination of Iron and Zinc in Hair and Human Serum Samples After Alkaline Solubilization by GF AAS

    OpenAIRE

    Donnici, Cláudio L.; Souza, Carolina C.; Beinner, Mark A.; Silva, José Bento B. da

    2016-01-01

    Methods for the development and validation for determination of iron and zinc in human serum and hair samples by graphite furnace atomic absorption spectrometry (GF AAS) were performed. Solubilization was immediate by manual agitation in both samples with a 10 mL volume sample plus deionized water. Optimum pyrolysis and atomization temperatures were obtained by pyrolysis and atomization temperature curves in both matrices. For serum samples, the best temperatures were 1400 and 2500 °C (Fe) an...

  15. Antimicrobial resistance in faecal samples from buffalo, wildebeest and zebra grazing together with and without cattle in Tanzania

    DEFF Research Database (Denmark)

    Katakweba, A. A. S.; Møller, K. S.; Muumba, J.

    2015-01-01

    = 40) and cattle (N = 20) from Mikumi National Park, Tanzania (MNP), where cattle is prohibited and from Ngorongoro Conservation Area (NCA) where co-grazing is practiced. The number of coliforms and enterococci resistant to selected antibiotics was determined. Wild life generally harboured higher...

  16. Comparative analysis of EV isolation procedures for miRNAs detection in serum samples

    Directory of Open Access Journals (Sweden)

    Zoraida Andreu

    2016-06-01

    Full Text Available Extracellular vesicles (EVs are emerging as potent non-invasive biomarkers. However, current methodologies are time consuming and difficult to translate to clinical practice. To analyse EV-encapsulated circulating miRNA, we searched for a quick, easy and economic method to enrich frozen human serum samples for EV. We compared the efficiency of several protocols and commercial kits to isolate EVs. Different methods based on precipitation, columns or filter systems were tested and compared with ultracentrifugation, which is the most classical protocol to isolate EVs. EV samples were assessed for purity and quantity by nanoparticle tracking analysis and western blot or cytometry against major EV protein markers. For biomarker validation, levels of a set of miRNAs were determined in EV fractions and compared with their levels in total serum. EVs isolated with precipitation-based methods were enriched for a subgroup of miRNAs that corresponded to miRNAs described to be encapsulated into EVs (miR-126, miR-30c and miR-143, while the detection of miR-21, miR-16-5p and miR-19a was very low compared with total serum. Our results point to precipitation using polyethylene glycol (PEG as a suitable method for an easy and cheap enrichment of serum EVs for miRNA analyses. The overall performance of PEG was very similar, or better than other commercial precipitating reagents, in both protein and miRNA yield, but in comparison to them PEG is much cheaper. Other methods presented poorer results, mostly when assessing miRNA by qPCR analyses. Using PEG precipitation in a longitudinal study with human samples, we demonstrated that miRNA could be assessed in frozen samples up to 8 years of storage. We report a method based on a cut-off value of mean of fold EV detection versus serum that provides an estimate of the degree of encapsulation of a given miRNA.

  17. Viral Nucleic Acids in the Serum Are Dependent on Blood Sampling Site in Patients with Clinical Suspicion of Myocarditis.

    Science.gov (United States)

    Pawlak, Agnieszka; Przybylski, Maciej; Durlik, Marek; Gil, Katarzyna; Nasierowska-Guttmejer, Anna M; Byczkowska, Katarzyna; Ziemba, Andrzej; Gil, Robert J

    2016-01-01

    The meaning of viral nucleic acids in the myocardium in many cases is difficult for clinical interpretation, whereas the presence of viral nucleic acids in the serum is a marker of active infection. We determined the diagnostic value of viral nucleic acids in ventricular serum and peripheral serum samples in comparison with endomyocardial biopsy (EMB) specimens in patients with clinically suspected myocarditis. The viral nucleic acid evaluation was performed in serum samples and EMB specimens by real-time PCR in 70 patients (age: 47 ± 16 years). The biopsy specimens were examined by histo- and immunohistochemistry to detect inflammatory response. The viral nucleic acids were detected in ventricular and peripheral serum, and EMB samples of 10 (14%), 14 (20%), and 32 (46%) patients, respectively. Notably, viral nucleic acids of the same virus as in the EMB sample were present more often in ventricular than in peripheral serum (60 vs. 7%, p = 0.01). A significant concurrence was observed between the positive and the negative results of viral nucleic acids present in EMB and ventricular serum (p = 0.0001). The detection of the same viral nucleic acid type in the myocardium and in ventricular serum being significantly more frequent than in the peripheral serum may suggest that the site of the blood collection is important for more precise and reliable confirmation of the active viral replication in the heart. © 2017 S. Karger AG, Basel.

  18. An attempt to validate serum and plasma as sample matrices for analyses of polychlorobiphenylols

    Energy Technology Data Exchange (ETDEWEB)

    Weiss, J.; Bergman, Aa. [Stockholm Univ. (Sweden). Dept. of Environmental Chemistry; Bignert, A. [Museum of Natural History (Sweden)

    2004-09-15

    Polychlorinated biphenyls (PCBs) form hydroxylated metabolites (OH-PCBs), as reported both from wildlife and from experimental animal studies already in the early 1970s'. However, the interest increased in OH-PCBs from the mid 1990s' depending on the discovery that some OHPCB congeners are strongly retained in the blood of birds, fish and mammals, including humans. The interest is linked to the fact that OH-PCBs is strongly, but reversibly, bound to the blood protein transthyretin (TTR). It is reasonable to believe that the strong TTR binding may have toxicological impact, probably related to endocrine type effects. Importantly, OH-PCBs are present in blood at far higher concentrations than in any other compartment in the body, which is dependent on the physico-chemical characteristics of the phenols. Analyses of OH-PCBs have thus been concentrated to whole blood, plasma or serum. Still there is no comparison between the three sample types even though it is clear that whole blood is not optimal due to the large proportion of haemoglobin in the sample that make the clean up more difficult than if plasma or serum is selected for analysis. In the present study we have addressed two questions: First we have looked at any potential differences in the analytical results of OH-PCBs when using serum and plasma for extraction and clean up; Second, the serum and plasma applied in the validation has been unfrozen, frozen (at -20 C) for two months and frozen for twenty months, respectively.

  19. Comparison of Milk Fat Globule Membrane (MFGM Proteins of Chianina and Holstein Cattle Breed Milk Samples Through Proteomics Methods

    Directory of Open Access Journals (Sweden)

    Lorraine Pariset

    2009-12-01

    Full Text Available Identification of proteins involved in milk production is important to understand the biology of lactation. Many studies have advanced the understanding of mammary function and milk secretion, but the critical molecular mechanisms implicated in milk fat secretion is still incomplete. Milk Fat Globules are secreted from the apical surface of the mammary cells, surrounded by a thin membrane bilayer, the Milk Fat Globule Membrane (MFGM, formed by proteins which have been suggested to be cholesterolemia-lowering factors, inhibitors of cancer cell growth, vitamin binders, bactericidal, suppressors of multiple sclerosis. Using a proteomic approach, we compared MFGM from milk samples of individuals belonging to two different cattle breeds, Chianina and Holstein, representative of selection for milk and meat traits, respectively. We were able to isolate some of the major MFGM proteins in the examined samples and to identify differences between the protein fractions of the two breeds. We detected differences in the amount of proteins linked to mammary gland development and lipid droplets formation, as well as host defence mechanisms. We have shown that proteomics is a suitable, unbiased method for the study of milk fractions proteins and a powerful tool in nutritional genomics.

  20. Stability of anti-HIV antibodies in serum samples stored for two to eighteen years periods

    Directory of Open Access Journals (Sweden)

    Marcia Jorge Castejon

    2014-08-01

    Full Text Available Introduction: The antibodies have an important role in the serodiagnosis, constituting the most widely used biomarkers to detect and confirm various diseases. Objective: To investigate the reproducibility of anti-human immunodeficiency virus (HIV antibodies reactivity, to assess the stability of the sera samples stored at -20ºC for two to eighteen years. Method: Sera were collected in the period 1988-2004 for routine anti-HIV antibodies diagnostic testing. The remaining samples stored at -20ºC, were analyzed in this study. Serum sample stability was assessed by enzyme-linked immunosorbent assay/enzyme immunoassay (ELISA/EIA, indirect immunofluorescence assay (IFA, and Western blot (WB for detecting anti-HIV antibodies. The previously found results (1988-2004 and those obtained in 2006 were subjected to Kappa index analysis. Result: In the period 1988-to 2004, the degree of concordance of the ELISA/EIA, IFA and WB results were considered, good (k = 0.80, regular (k = 0.35, and good (k = 0.63, respectively. Conclusion: Regarding HIV serologic test, the serum samples were stable for 18 years in ELISA/EIA and for 4 years in IFA technique, however, for the WB methodology it was not possible to determine the time of stability of the anti-HIV antibodies.

  1. Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system.

    Science.gov (United States)

    Arévalo, Fernando Javier; Messina, Germán Alejandro; Molina, Patricia Gabriela; Zón, María Alicia; Raba, Julio; Fernández, Héctor

    2010-03-15

    Progesterone (P4) is a steroidal hormone with a vital role in the maintenance of human and animal health. This paper describes the development of an immunosensor coupled to glassy carbon (GC) electrode and integrated to a microfluidic system to quantify P4 from bovine serum samples in a fast and sensitive way. The serum samples spiked with a given P4 concentration and a given P4 concentration bound to horseradish peroxide (HPR) were simultaneously added and, therefore, they competed immunologically with sheep monoclonal anti-P4 antibodies that were immobilized at a rotating disk. HRP in the presence of hydrogen peroxide (H(2)O(2)) catalyzes the chatecol (H(2)Q) oxidation to benzoquinone (Q). Its reverse electrochemical reduction to H(2)Q can be detected at a GC electrode surface at -0.15 V by chronoamperometric measurements. These current responses are proportional to the enzyme activity and inversely proportional to the P4 amount present in bovine serum samples. This P4 immunosensor showed a linear working range from 0.5 to 12.5 ng mL(-1). The detection (DL) and quantification (QL) limits were 0.2 and 0.5 ng mL(-1), respectively. The electrochemical immunosensor had a higher sensitivity than the ELISA method using conventional spectrophotometric detections. However, both methods allowed us to obtain similar detection limits. The immunosensor allowed us to make up to 100 determinations on different samples without any previous pre-treatment. This behavior proved to be suitable to detect P4 in routine veterinary, clinical, biological, physiological, and analytical assays. Copyright (c) 2009 Elsevier B.V. All rights reserved.

  2. Application of PCR in serum samples for diagnosis of paracoccidioidomycosis in the southern Bahia-Brazil.

    Directory of Open Access Journals (Sweden)

    Lucas Dias

    Full Text Available Paracoccidioidomycosis (PCM cannot always be diagnosed by conventional means such as direct examination of histopathology or clinical samples, and serological methods, used as an alternative, still have many cases of cross-reactivity. In this scenario, molecular techniques seem to arise as a rapid approach, specific and direct that could be used in the diagnosis of this mycosis. In this study we analyzed 76 serum samples from patients in southern Bahia suspected of having paracoccidioidomycosis using a conventional PCR with primers for the ITS1 ribosomal DNA of P. brasiliensis. Of these 76 patients, 5 were positive for PCM by double immunodiffusion and/or direct examination and histopathology. To test specificity of PCR, we used human DNA and three isolates of P. lutzii (1578, 01 and ED01. Additionally, we analyzed by serial dilutions of DNA the limit of detection of the assay. The test of PCR proved specific, as only a 144 bp fragment of the three isolates of P. lutzii and no human DNA was amplified. Detection limit was 1.1 pg/µL of DNA. Despite the high detection limit and specificity of PCR none of the 76 serum samples were found positive by PCR, but a biopsy specimen obtained from one of the patients with PCM was positive. These results, albeit limited, show that PCR is not effective in detecting DNA of P. brasiliensis or P. lutzii in serum, but could perhaps be used with other types of clinical samples, especially in those instances in which conventional methods fail.

  3. Comparison of three techniques in the preparation of samples for the crystallization of cervical flow in lactating dairy cattle

    Directory of Open Access Journals (Sweden)

    Reátegui J

    2017-08-01

    Full Text Available The objective was to compare three techniques of sample preparation for cervical flow crystallization (pressure imprint, touch imprint and smear, analyzing the tree forms (crystallization as a characterization of the cervical flow of dairy cattle, according to the day of collection and moment of the estrous cycle. Ten clinically healthy, multiparous Holstein Friesian dairy cows were sampled and 30 to 50 days postpartum. Each was collected from the vaginal cervix using a disposable pipette and a 50cc syringe. The two imprints and the smear were prepared on slides, with two-step protocol according to the methodology of Prado et al. (2012. The samples were then allowed to dry in the environment and the microscope was read with a higher magnification objective (40X to observe the formation of the crystals, these procedures were performed in 4 different moments of the estrous cycle (0, 7, 14, 21 days. To quantify the crystallization, a scale from 0 to 4 was used, which varies depending on the formation of typical crystals at least formation or absence. At day 0, 7, 14, 21, the crystallization level in the three techniques had significant difference (P <0.05. At day 0, 50% of the samples processed by the touch imprint and pressure imprint showed typical formation compared to 20% that were processed by the smear technique. On day 7, 80% of the samples processed by touch imprint, 90% of the smear technique and 70% of the pressure imprint, present atypical crystals. On day 14, 60% of the samples processed by the contact imprint and 30% and 40% of the samples processed by the smear technique and pressure imprint, respectively, showed atypical crystal formation. On day 21, 40% of the samples processed by the touch imprint and 10% of the samples processed by the smear technique and pressure imprint showed typical crystal formation. It is concluded that the techniques of preparation of samples influence the crystallization of cervical mucus, being the most

  4. Amperometric immunosensor for the determination of IgA deficiency in human serum samples.

    Science.gov (United States)

    Rosales-Rivera, Luis Carlos; Acero-Sánchez, Josep Lluis; Lozano-Sánchez, Pablo; Katakis, Ioanis; O'Sullivan, Ciara K

    2012-03-15

    An electrochemical immunosensor for the detection of human IgA deficiency in real human blood serum has been developed. The performance of the immunosensor presents a large but sensitive dynamic range that allows the determination of non-deficient IgA levels (>70 μg/mL) as well as of severe IgA deficiencies (0.5-5.0 μg/mL). The assay architecture involves the immobilisation of a coating antibody on an electrode surface using carboxylic-ended bipodal alkane-thiol self-assembled monolayers (SAMs). The long chain bipodal SAM presents intercalated poly(ethylenglycol) groups that confer the immunosensor the ability to retain its optimum performance in very complex matrices and serum with negligible non-specific adsorption phenomena. Amperometric optimisation of the assay resulted in limits of detection of 142 ng/mL in just 30 min total assay time. Real patients' serum samples were analysed using the developed electrochemical immunosensor demonstrating an excellent correlation in terms of sensitivity and reproducibility compared with standard enzyme linked immunosorbent assays (ELISA). Copyright © 2012 Elsevier B.V. All rights reserved.

  5. Specific Antibodies Reacting with SV40 Large T Antigen Mimotopes in Serum Samples of Healthy Subjects.

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    Mauro Tognon

    Full Text Available Simian Virus 40, experimentally assayed in vitro in different animal and human cells and in vivo in rodents, was classified as a small DNA tumor virus. In previous studies, many groups identified Simian Virus 40 sequences in healthy individuals and cancer patients using PCR techniques, whereas others failed to detect the viral sequences in human specimens. These conflicting results prompted us to develop a novel indirect ELISA with synthetic peptides, mimicking Simian Virus 40 capsid viral protein antigens, named mimotopes. This immunologic assay allowed us to investigate the presence of serum antibodies against Simian Virus 40 and to verify whether Simian Virus 40 is circulating in humans. In this investigation two mimotopes from Simian Virus 40 large T antigen, the viral replication protein and oncoprotein, were employed to analyze for specific reactions to human sera antibodies. This indirect ELISA with synthetic peptides from Simian Virus 40 large T antigen was used to assay a new collection of serum samples from healthy subjects. This novel assay revealed that serum antibodies against Simian Virus 40 large T antigen mimotopes are detectable, at low titer, in healthy subjects aged from 18-65 years old. The overall prevalence of reactivity with the two Simian Virus 40 large T antigen peptides was 20%. This new ELISA with two mimotopes of the early viral regions is able to detect in a specific manner Simian Virus 40 large T antigen-antibody responses.

  6. Simpler and Faster Spectrophotometric Determination of Diclofenac Sodium in Tablets, Serum and Urine Samples

    Directory of Open Access Journals (Sweden)

    Abdul Rauf Khaskheli

    2009-12-01

    Full Text Available This work describes a simple, sensitive, rapid and economical analytical procedure for direct spectrophotometric evaluation of diclofenac sodium (DS using aqueous medium without using a chemical reagent. Parameters like time, temperature, acidic and basic conditions and interference by analgesic drugs were studied for a 5µg ml-1 solution of DS at 276 nm. Under optimized parameters, a linear working range of 0.1–30 g ml-1 with regression coefficient of 0.9998 and lower detection limit of 0.01 g ml-1 was obtained. The method was applied for DS contents in tablets, serum and urine samples.

  7. Serological and molecular detection of bovine leukemia virus in cattle in Iraq

    Science.gov (United States)

    Khudhair, Yahia Ismail; Hasso, Saleem Amin; Yaseen, Nahi Y; Al-Shammari, Ahmed Majeed

    2016-01-01

    Bovine leukemia virus (BLV) is highly endemic in many countries, including Iraq, and it impacts the beef and dairy industries. The current study sought to determine the percentage of BLV infection and persistent lymphocytosis (PL) in cattle in central Iraq. Hematological, serological, and molecular observations in cross breeds and local breeds of Iraqi cattle naturally infected with BLV were conducted in the peripheral blood mononuclear cells of 400 cattle (340 cross breed and 60 local breed) using enzyme-linked immunosorbent assay and polymerase chain reaction (PCR). On the basis of the absolute number of lymphocytes, five of the 31 positive PCR cases had PL. Among these leukemic cattle, one case exhibited overt neutrophilia. Serum samples were used to detect BLV antibodies, which were observed in 28 (7%) samples. PCR detected BLV provirus in 31 samples (7.75%). All 28 of the seropositive samples and the 3 seronegative samples were positive using PCR. Associations were observed between bovine leukosis and cattle breed, age and sex. Age-specific analysis showed that the BLV percentage increased with age in both breeds. Female cattle (29 animals; 7.34%) exhibited significantly higher infectivity than male cattle (two animals; 4.34%). In conclusion, comprehensive screening for all affected animals is needed in Iraq; programs that segregate cattle can be an effective and important method to control and/or eliminate the BLV. PMID:27273225

  8. Prevalence of Coxiella burnetii in environmental samples collected from cattle farms in Eastern and Central Poland (2011-2012).

    Science.gov (United States)

    Bielawska-Drózd, Agata; Cieślik, Piotr; Mirski, Tomasz; Gaweł, Jerzy; Michalski, Aleksander; Niemcewicz, Marcin; Bartoszcze, Michał; Żakowska, Dorota; Lasocki, Krzysztof; Knap, Józef; Kocik, Janusz

    2014-12-05

    Coxiella burnetii is the etiologic agent of Q fever. It may occur as two different morphological forms, a large cell variant (LCV) and a small cell variant (SCV). The SCV is characterized by unique resistance to physical and chemical factors and may survive in the environment for many months. The objective of this study was to examine environmental samples for the presence of C. burnetii using real-time PCR in areas where Q fever was previously reported and in randomly selected animal farms where Q fever was not reported. The samples were collected in the following provinces in Poland: Lublin, Subcarpathian and Masovian. Monitoring was performed with real-time PCR and serological methods. Of the 727 environmental samples, 33 (4.54%) contained the multi-copy insertion sequence IS1111, which is specific for C. burnetii. Subsequently, the presence of C. burnetii antibodies was determined using serological tests in selected herds in which positive genetic results were obtained. Serological analyses of 169 serum samples using CFT and ELISA were performed on Polish black-and-white Holstein-Friesian cows and one cow imported from Denmark. Using the CFT method, 11 samples were positive for phase I antibodies and six were positive for phase II antibodies. Moreover, in two cases, the presence of antibodies specific for both phase I and phase II antigens of C. burnetii was detected. However, of the 169 examined serum samples, 20 were positive by ELISA test, of which six were also positive by CFT. Additionally, multi spacer typing (MST) of isolated C. burnetii strains was performed. The MST results identified two new genotypes in Poland, ST3 and ST6. The results indicate that continued research regarding spread of this pathogen within a country is necessary. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

  9. Applications of magnetic surface imprinted materials for solid phase extraction of levofloxacin in serum samples.

    Science.gov (United States)

    Xiao, Deli; Wang, Cuixia; Dai, Hao; Peng, Jun; He, Jia; Zhang, Kai; Kong, Sumei; Qiu, Panzi; He, Hua

    2015-05-01

    In this work, molecularly imprinted magnetic carbon nanotubes (MCNTs@MIPs) was prepared with surface imprinting technique for extraction of levofloxacin in serum samples. The preparation of molecularly imprinted polymers (MIPs) used levofloxacin as template, methacrylic acid as functional monomer, and ethylene glycol dimethacrylate as cross-linker, and the magnetic carbon nanotubes (MCNTs) was synthesized by solvothermal method. The prepared polymers not only can be separated and collected easily by an external magnetic, but also exhibited high specific surface area and high selectivity to template molecules. Kinetic adsorption and static adsorption capacity investigations indicated that the synthesized MCNTs@MIPs had excellent recognition towards levofloxacin. Furthermore, magnetic solid phase extraction (MSPE) using the prepared MCNTs@MIPs as sorbent was then investigated, and an efficient sample cleanup was obtained with recoveries ranged from 78.7 ± 4.8 % to 83.4 ± 4.1%. In addition, several parameters, including the pH of samples, the amount of MCNTs@MIPs, the adsorption and desorption times, and the eluent, were investigated to obtain optimal extraction efficiency. Under the optimal extraction conditions, the stability of the polymer was also evaluated, and the average recovery reduced less than 7.6% after 5 cycles. MCNTs@MIPs successfully applied in the preconcentration and determination of levofloxacin in serum sample suggested that the MSPE method based on the novel polymers could be a promising alternative for selective and efficient extraction of trace amounts of pharmaceutical substances in bio-matrix samples. Copyright © 2015 John Wiley & Sons, Ltd.

  10. Chronic arsenicosis in cattle: possible mitigation with Zn and Se.

    Science.gov (United States)

    Dash, Jeevan R; Datta, Bakul K; Sarkar, Samar; Mandal, Tapan K

    2013-06-01

    Supply of arsenic free water in the arsenic endemic zone of West Bengal since a long time could not prevent arsenicosis in human completely. So exploration of arsenic contamination at all levels of food chain may be important. The report on arsenicosis in cattle of arsenic affected zone is scarcely available. In the present study, cattle from villages of two arsenic endemic blocks (Chakdah and Haringhata) in Nadia district of West Bengal were selected. The cattle affected with arsenicosis were identified and isolated. They were divided into two groups: untreated control group and treated group-was treated with mixture of zinc oxide (10mg/kg) and sodium selenite (0.1mg/kg) orally once daily for 180 days. Milk, faeces and hair samples were collected at different time intervals from those cattle for analysis of arsenic. Drinking water and straw of those areas were also collected for analysis of arsenic. Serum ALT and AST were estimated in collected blood samples of the cattle. The untreated control group had shown gradual accumulation of total arsenic in hair while the treated group showed a non-significant but declined accumulation of arsenic in hair from 90th day onwards and a significant increase (p<0.05) in total arsenic in faeces from 90th day onwards. The arsenic load considerably but non-significantly decreased in milk from 60th day onwards in the treated group. Serum AST and ALT activities were also decreased in treated cattle. It is concluded that straw is also a major source of arsenic contamination in cattle apart from drinking water and arsenic may enter into human through consumption of contaminated milk. But zinc and selenium mixture may be used to reduce arsenic load in cattle. Copyright © 2013 Elsevier Inc. All rights reserved.

  11. Direct Assessment of Plasma/Serum Sample Quality for Proteomics Biomarker Investigation.

    Science.gov (United States)

    Greco, Viviana; Piras, Cristian; Pieroni, Luisa; Urbani, Andrea

    2017-01-01

    Blood proteome analysis for biomarker discovery represents one of the most challenging tasks to be achieved through clinical proteomics due to the sample complexity, such as the extreme heterogeneity of proteins in very dynamic concentrations, and to the observation of proper sampling and storage conditions. Quantitative and qualitative proteomics profiling of plasma and serum could be useful both for the early detection of diseases and for the evaluation of pathological status. Two main sources of variability can affect the precision and accuracy of the quantitative experiments designed for biomarker discovery and validation. These sources are divided into two categories, pre-analytical and analytical, and are often ignored; however, they can contribute to consistent errors and misunderstanding in biomarker research. In this chapter, we review critical pre-analytical and analytical variables that can influence quantitative proteomics. According to guidelines accepted by proteomics community, we propose some recommendations and strategies for a proper proteomics analysis addressed to biomarker studies.

  12. Evaluation of a new antibody-based enzyme-linked immunosorbent assay for the detection of bovine leukemia virus infection in dairy cattle

    NARCIS (Netherlands)

    Monti, G.E.; Frankena, K.; Engel, B.; Buist, W.; Tarabla, H.D.; Jong, de M.C.M.

    2005-01-01

    The objective of this study was to validate a new blocking enzyme-linked immunosorbent assay (ELISA) (designated M108 for milk and S108 for serum samples) for detecting bovine leukemia virus (BLV) infection in dairy cattle. Milk, serum, and ethylenediaminetetraacetic acid-blood samples were

  13. Studies on Using Cattle and Sheep Hydatid Cyst Fluid Instead of the Fetal Calf Serum in Leishmania Culture

    Directory of Open Access Journals (Sweden)

    Hossein Rezvan

    2013-12-01

    Full Text Available Background: Leishmania is a single cell parasite causing leishmaniasis, which is a common disease between humans and animals. Due to the importance of in-vitro culture of the parasite in leishmania research, developing new methods for in-vitro cultivation of the parasite has always been a goal for leishmania researchers. The main objective of7T 5T7Tthis study was to use sheep and bovine hydatid cyst fluids as alternatives for fetal calf serum (FCS in leishmania in-vitro5T culture5T. Materials and Methods: 7TA total of 5T7T1 million leishmania promastigotes were added to 4 flasks as follow5T7T. A f5T7Tlask containing DMEM medium with 105T7T% 5T7Tfetal bovine serum5T7T, a f5T7Tlask containing DMEM and 10% sheep hydatid cyst fluid5T7T, a f5T7Tlask containing DMEM medium with 105T7T% 5T7Tbovine hydatid cyst fluid and a5T7T f5T7Tlask containing DMEM medium alone. After 2, 45T7T, 5T7T7, 95T7T, 11, 5T7T21 and 24 days, the number of parasites were counted and compared5T7T. Results: The result of this study showed that, DMEM medium enriched with 10% sheep hydatid cyst fluid in 168 hours and medium enriched with 10% bovine hydatid cyst fluid in 96 hours can act as a good alternative for fetal bovine serum in the culture Leishmania major. Conclusion: 5TThe results showed that sheep and bovine hydatid cyst fluid can be used as alternatives to FCS for dense cultivation of leishmania. The results also showed that5T, 5Tthe growth of promastigotes in medium enriched with bovine cyst fluid is more rapid than the medium enriched with sheep5T c5Tyst fluid5T in5T the beginning of cultivation.

  14. Technical note: Analysis of total lipid and triacylglycerol content in small liver biopsy samples in cattle.

    Science.gov (United States)

    Starke, A; Haudum, A; Busche, R; Beyerbach, M; Dänicke, S; Rehage, J

    2010-08-01

    A procedure is described for analyzing total lipid (TL) and triacylglycerol (TAG) in 2 sequential steps using small amounts (TAG was measured enzymatically in the TL extract, using an automated analyzer. For gravimetric TL determination in milligrams per gram of liver fresh weight (FW), TL was extracted from homogenized tissue samples with hexane:isopropanol (at 20 degrees C, 24 h, constant agitation). The routine method was modified by adding a second hexane extraction step to optimize lipid extraction. The dry lipid extract was dissolved in hexane and aliquoted according to TL content for TAG analysis. An extra incubation period of 16 h was included for complete hydrolysis of TAG, using microbial lipase and nonaethylene glycol monododecyl ether detergent, before TAG was measured enzymatically using commercial test kits. Triolein was used as an internal standard. Repeated TL analysis (n = 3) of liver specimens from 10 cows (range, 40 to 314 mg/g of FW) yielded a mean CV of 2.2%, whereas repeated TAG analysis (range, 4 to 260 mg/g of FW) yielded a mean intraday CV of 2.5% (n = 5) and a mean interday CV of 3.4% (n = 4). Intraday (n = 5) and interday (n = 4) CV for repeated TAG analysis in triolein standards were TAG in triolein standards varied between 99 and 103%. In part 2 of the experiment, hepatic TL and TAG were measured in 150 German Holstein cows to verify the test method in a large sample size. For repeated hepatic TL (n = 3) and TAG (n = 5) determination, mean CV of TAG relative to TL increased linearly to a breakpoint of approximately 100 mg TL/g of FW, at which point it reached a plateau at approximately 68%, indicating an accumulation of other lipid fractions in hepatic tissue with hepatic TL above the breakpoint. Calculation of hepatic TAG from TL was reasonably accurate when a 2-slope linear broken-line model (r(2) = 0.98) was used. Above a TL of approximately 40 mg/g of FW, calculated TAG values deviated by only +/-15% from measured hepatic TAG.

  15. Development of a Spatially Targeted Field Sampling Technique for the Southern Cattle Tick, Rhipicephalus microplus, by Mapping Whitetailed Deer, Odocoileus virginianus, Habitat in South Texas

    OpenAIRE

    Phillips, Pamela L.; Welch, John B.; Kramer, Matthew

    2014-01-01

    The objective of our study was to determine whether satellite remote sensed data could be used to identify white-tailed deer, Odocoileus virginianus (Zimmerman) (Artiodactyla: Cervidae), habitat and target locations for sampling free-living larvae of the southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini) (Ixodida: Ixodidae) in South Texas. Two methods for mapping white-tailed deer habitat were used, an object-oriented method to identify closed canopies and waterways for de...

  16. Serum Neutralization Assay Can Efficiently Replace Plaque Reduction Neutralization Test for Detection and Quantitation of West Nile Virus Antibodies in Human and Animal Serum Samples

    Science.gov (United States)

    Di Gennaro, Annapia; Casaccia, Claudia; Conte, Annamaria; Monaco, Federica; Savini, Giovanni

    2014-01-01

    A serum neutralization assay (SN) was compared with the official plaque reduction neutralization test for the quantitation of West Nile virus antibodies. A total of 1,348 samples from equid sera and 38 from human sera were tested by these two methods. Statistically significant differences were not observed, thus supporting the use of SN for routine purposes. PMID:25100824

  17. The detection of lumpy skin disease virus in samples of experimentally infected cattle using different diagnostic techniques

    Directory of Open Access Journals (Sweden)

    E.S.M. Tuppurainen

    2005-09-01

    Full Text Available Lumpy skin disease (LSD is a disease of cattle, primarily in Africa and Madagascar and rarely in the Middle East. It is caused by a capripoxvirus that belongs to the family Poxviridae. The disease is of economic importance in endemic areas. Effective control of LSD requires accurate and rapid laboratory techniques to confirm a tentative clinical diagnosis. Comparative studies on different diagnostic tests used at different stages of the disease have not been done. The aim of this study was to compare several of these tests. Six seronegative bulls, between 11 and 20 months of age, were infected intravenously and kept in an insect-free facility. The course of the infection was monitored. During a 3-month period blood samples and skin biopsies were collected for virus isolation and polymerase chain reaction (PCR. Skin biopsies were also examined using transmission electron microscopy (TEM. The incubation period in infected animals varied from 4-5 days. The length of the viraemic period did not correlate with the severity of clinical disease. Viraemia was detected from 1-12 days using virus isolation and from 4-11 days using the PCR, which is longer than has previously been reported. Virus was isolated from skin biopsies until Day 39 post infection (p.i. and PCR could demonstrate viral DNA until Day 92 p.i. Transmission electron microscopy of negatively stained skin biopsies detected LSD virus only in one of the four bulls that developed skin lesions until Day 33 p.i. The PCR was a fast and sensitive method to demonstrate viral DNA in blood and skin samples. It could detect viral nucleic acid in skin lesions 53 days longer than virus isolation. Virus isolation from blood and skin samples was sensitive and reliable, but as a single test it may be too time-consuming to use although this depends on how rapidly the diagnosis must be confirmed. In conclusion, this study showed the PCR to be superior in detecting LSD virus from blood and skin samples

  18. Liquid Chromatographic Determination of Pioglitazone in Pharmaceuticals, Serum and Urine Samples

    Directory of Open Access Journals (Sweden)

    Shahnaz Perveen

    2011-12-01

    Full Text Available A rapid and reliable analytical method based on high-performance liquid chromatography (HPLC with UV detection (221nm has been developed for the determination of the anti-hyperglycemic agent Pioglitazone in pharmaceutical formulations and biological fluids (serum and urine after clean-up with solid-phase extraction. Chromatographic separation was achieved with a Chromolith® Performance RP-18e (100×4.6mm column using mobile phase composition of acetonitrile: mixed phosphate buffer (pH 2.5; 10mM (30:70, v/v with a flow rate of 2.0mL/min. The total run time was 2 min. under optimized conditions. The calibration curve was found to be linear in the range of 1-10 µg mL-1 with regression coefficient of 0.9996, and the lower limit of detection 72 ng/20µL injection. The method has been validated for the system suitability, linearity, precision and accuracy, limits of detection, specificity, stability and robustness. The %recovery of Pioglitazone in pharmaceutical formulations was found to be 104.7%. The assay has been applied successfully to the pharmaceutical Tablet samples and biological fluids (serum and urine of healthy volunteers.

  19. Milk and serum standard reference materials for monitoring organic contaminants in human samples

    Science.gov (United States)

    Eppe, Gauthier; Focant, Jean-François; Hamilton, Coreen; Heckert, N. Alan; Heltsley, Rebecca M.; Hoover, Dale; Keller, Jennifer M.; Leigh, Stefan D.; Patterson, Donald G.; Pintar, Adam L.; Sharpless, Katherine E.; Sjödin, Andreas; Turner, Wayman E.; Vander Pol, Stacy S.; Wise, Stephen A.

    2016-01-01

    Four new Standard Reference Materials (SRMs) have been developed to assist in the quality assurance of chemical contaminant measurements required for human biomonitoring studies, SRM 1953 Organic Contaminants in Non-Fortified Human Milk, SRM 1954 Organic Contaminants in Fortified Human Milk, SRM 1957 Organic Contaminants in Non-Fortified Human Serum, and SRM 1958 Organic Contaminants in Fortified Human Serum. These materials were developed as part of a collaboration between the National Institute of Standards and Technology (NIST) and the Centers for Disease Control and Prevention (CDC) with both agencies contributing data used in the certification of mass fraction values for a wide range of organic contaminants including polychlorinated biphenyl (PCB) congeners, chlorinated pesticides, polybrominated diphenyl ether (PBDE) congeners, and polychlorinated dibenzo-p-dioxin (PCDD) and dibenzofuran (PCDF) congeners. The certified mass fractions of the organic contaminants in unfortified samples, SRM 1953 and SRM 1957, ranged from 12 ng/kg to 2200 ng/kg with the exception of 4,4′-DDE in SRM 1953 at 7400 ng/kg with expanded uncertainties generally <14 %. This agreement suggests that there were no significant biases existing among the multiple methods used for analysis. PMID:23132544

  20. Comparing identified and statistically significant lipids and polar metabolites in 15-year old serum and dried blood spot samples for longitudinal studies: Comparing lipids and metabolites in serum and DBS samples

    Energy Technology Data Exchange (ETDEWEB)

    Kyle, Jennifer E. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Casey, Cameron P. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Stratton, Kelly G. [National Security Directorate, Pacific Northwest National Laboratory, Richland WA USA; Zink, Erika M. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Kim, Young-Mo [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Zheng, Xueyun [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Monroe, Matthew E. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Weitz, Karl K. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Bloodsworth, Kent J. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Orton, Daniel J. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Ibrahim, Yehia M. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Moore, Ronald J. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Lee, Christine G. [Department of Medicine, Bone and Mineral Unit, Oregon Health and Science University, Portland OR USA; Research Service, Portland Veterans Affairs Medical Center, Portland OR USA; Pedersen, Catherine [Department of Medicine, Bone and Mineral Unit, Oregon Health and Science University, Portland OR USA; Orwoll, Eric [Department of Medicine, Bone and Mineral Unit, Oregon Health and Science University, Portland OR USA; Smith, Richard D. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Burnum-Johnson, Kristin E. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Baker, Erin S. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA

    2017-02-05

    The use of dried blood spots (DBS) has many advantages over traditional plasma and serum samples such as smaller blood volume required, storage at room temperature, and ability for sampling in remote locations. However, understanding the robustness of different analytes in DBS samples is essential, especially in older samples collected for longitudinal studies. Here we analyzed DBS samples collected in 2000-2001 and stored at room temperature and compared them to matched serum samples stored at -80°C to determine if they could be effectively used as specific time points in a longitudinal study following metabolic disease. Four hundred small molecules were identified in both the serum and DBS samples using gas chromatograph-mass spectrometry (GC-MS), liquid chromatography-MS (LC-MS) and LC-ion mobility spectrometry-MS (LC-IMS-MS). The identified polar metabolites overlapped well between the sample types, though only one statistically significant polar metabolite in a case-control study was conserved, indicating degradation occurs in the DBS samples affecting quantitation. Differences in the lipid identifications indicated that some oxidation occurs in the DBS samples. However, thirty-six statistically significant lipids correlated in both sample types indicating that lipid quantitation was more stable across the sample types.

  1. Stability of BDNF in Human Samples Stored Up to 6 Months and Correlations of Serum and EDTA-Plasma Concentrations.

    Science.gov (United States)

    Polyakova, Maryna; Schlögl, Haiko; Sacher, Julia; Schmidt-Kassow, Maren; Kaiser, Jochen; Stumvoll, Michael; Kratzsch, Jürgen; Schroeter, Matthias L

    2017-06-03

    Brain-derived neurotrophic factor (BDNF), an important neural growth factor, has gained growing interest in neuroscience, but many influencing physiological and analytical aspects still remain unclear. In this study we assessed the impact of storage time at room temperature, repeated freeze/thaw cycles, and storage at -80 °C up to 6 months on serum and ethylenediaminetetraacetic acid (EDTA)-plasma BDNF. Furthermore, we assessed correlations of serum and plasma BDNF concentrations in two independent sets of samples. Coefficients of variations (CVs) for serum BDNF concentrations were significantly lower than CVs of plasma concentrations (n = 245, p = 0.006). Mean serum and plasma concentrations at all analyzed time points remained within the acceptable change limit of the inter-assay precision as declared by the manufacturer. Serum and plasma BDNF concentrations correlated positively in both sets of samples and at all analyzed time points of the stability assessment (r = 0.455 to rs = 0.596; p < 0.004). In summary, when considering the acceptable change limit, BDNF was stable in serum and in EDTA-plasma up to 6 months. Due to a higher reliability, we suggest favoring serum over EDTA-plasma for future experiments assessing peripheral BDNF concentrations.

  2. Stability of BDNF in Human Samples Stored Up to 6 Months and Correlations of Serum and EDTA-Plasma Concentrations

    Directory of Open Access Journals (Sweden)

    Maryna Polyakova

    2017-06-01

    Full Text Available Brain-derived neurotrophic factor (BDNF, an important neural growth factor, has gained growing interest in neuroscience, but many influencing physiological and analytical aspects still remain unclear. In this study we assessed the impact of storage time at room temperature, repeated freeze/thaw cycles, and storage at −80 °C up to 6 months on serum and ethylenediaminetetraacetic acid (EDTA-plasma BDNF. Furthermore, we assessed correlations of serum and plasma BDNF concentrations in two independent sets of samples. Coefficients of variations (CVs for serum BDNF concentrations were significantly lower than CVs of plasma concentrations (n = 245, p = 0.006. Mean serum and plasma concentrations at all analyzed time points remained within the acceptable change limit of the inter-assay precision as declared by the manufacturer. Serum and plasma BDNF concentrations correlated positively in both sets of samples and at all analyzed time points of the stability assessment (r = 0.455 to rs = 0.596; p < 0.004. In summary, when considering the acceptable change limit, BDNF was stable in serum and in EDTA-plasma up to 6 months. Due to a higher reliability, we suggest favoring serum over EDTA-plasma for future experiments assessing peripheral BDNF concentrations.

  3. Pine needle abortion in cattle update: Metabolite detection in sera and fetal fluids from abortion case samples

    Science.gov (United States)

    Cattle abortions associated with consumption of pine needles during late gestation are a serious poisonous plant problem in the Western US. Most cases of abortion have been associated with consumption of ponderosa pine (Pinus ponderosa) and the causative agent was identified as the labdane diterpen...

  4. Development of a spatially targeted field sampling technique for the southern cattle tick, Rhipicephalus microplus, by mapping white-tailed deer, Odocoileus virginianus, habitat in South Texas.

    Science.gov (United States)

    Phillips, Pamela L; Welch, John B; Kramer, Matthew

    2014-01-01

    The objective of our study was to determine whether satellite remote sensed data could be used to identify white-tailed deer, Odocoileus virginianus (Zimmerman) (Artiodactyla: Cervidae), habitat and target locations for sampling free-living larvae of the southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini) (Ixodida: Ixodidae) in South Texas. Two methods for mapping white-tailed deer habitat were used, an object-oriented method to identify closed canopies and waterways for deer movement and two vegetation indices: the Normalized Difference Vegetation Index and the Modified Soil Adjusted Vegetation Index to identify forage for deer. These two data sets of favorable white-tailed deer habitat were combined within a geographic information system to identify locations for sampling ticks. Larvae of R. (B.) microplus, were sampled in Zapata County, Texas, by walking transects with attached flannel panels to jeans. Although the data set and sampling period were limited, data analysis demonstrated that sampling of free-living larvae of R. (B.) microplus can be conducted in South Texas, and larvae were most abundant in areas that harbored O. virginianus. Spatial analysis of satellite imagery to classify white-tailed deer/southern cattle tick habitat proved efficacious and may be useful in directing sampling activities in the field. This is an open access paper. We use the Creative Commons Attribution 3.0 license that permits unrestricted use, provided that the paper is properly attributed.

  5. Effects of coagulation temperature on measurements of complement function in serum samples from patients with systemic lupus erythematosus

    DEFF Research Database (Denmark)

    Baatrup, G; Sturfelt, G; Junker, A

    1992-01-01

    Blood samples from 15 patients with systemic lupus erythematosus (SLE) and 15 healthy blood donors were allowed to coagulate for one hour at room temperature, followed by one hour at 4 or 37 degrees C. The complement activity of the serum samples was assessed by three different functional assays...

  6. Retrospective analysis of dengue specific IgM reactive serum samples

    Directory of Open Access Journals (Sweden)

    Nemai Bhattacharya

    2013-04-01

    Full Text Available Objective: To conduct a retrospective analysis of dengue cases in Kolkata, on the basis of presence of anti-dengue IgM in their sera and presence or absence of anti-dengue IgG and dengue specific Non structural 1 (NS1 antigen in each of the serum sample. Methods: Sample was tested quantitatively employing ELISA technique, using Biorad test kits, with a view to get a more comprehensive picture of dengue in an urban endemic area and also to evaluate individual cases. Results: This reconstructed study revealed that of those 91 dengue cases, 70.3% (64 and 29.7% (27 were suffering from secondary and primary dengue respectively, showing that number of secondary dengue cases were much more than that of primary dengue cases with a possibility of emergence of DHF. A small proportion of cases 18.7% (17 were reactive for NS1. The duration of fever in NS1 antigen positive cases varied between 5 and 7 days. Of 17 NS1 reactive cases, 10 (10.9% and 7 (7.7% were suffering from secondary and primary dengue respectively. Conclusions: Early detection of primary and secondary dengue cases would be facilitated by utilizing all three parameters (NS1 antigen, anti-dengue IgM and IgG helping to evaluate, monitor and treat a dengue case effectively.

  7. Development of a diagnostic method for neosporosis in cattle using recombinant Neospora caninum proteins.

    Science.gov (United States)

    Dong, Jinhua; Otsuki, Takahiro; Kato, Tatsuya; Park, Enoch Y

    2012-05-04

    Neosporosis is an infectious disease primarily of cattle and dogs, caused by intracellular parasite, Neospora caninum. Neosporosis appears to be a major cause of abortion in dairy cattle worldwide and causes to huge economic loss to dairy industry. Recombinant surface associated antigen 1 (NcSAG1), NcSAG1 related sequence 2 (NcSRS2) and the dense granule antigen 2 (NcGRA2) of N. caninum were expressed either in silkworm or in Escherichia coli and purified. The purified recombinant proteins bound to the N. caninum-specific antibodies in serum samples from infected cattle as revealed by an enzyme-linked immunosorbent assay (ELISA). By co-immobilizing these recombinant proteins, a novel indirect ELISA was developed for detection of neosporosis. With the use of 32 serum samples, comprising 12 positive serum samples and 20 negative serum samples, the sensitivity and specificity of the assay were found to be 91.7 and 100%, respectively. Seventy-two serum samples from dairy farms were also tested and one was diagnosed with neosporasis with both this method and a commercial assay. A diagnostic method employing recombinant proteins of N. caninum was developed. The method showed high sensitivity and specificity. Diagnostic test with field serum samples suggested its applicability to the practical diagnosis of neosporosis.

  8. Development of a diagnostic method for neosporosis in cattle using recombinant Neospora caninum proteins

    Directory of Open Access Journals (Sweden)

    Dong Jinhua

    2012-05-01

    Full Text Available Abstract Background Neosporosis is an infectious disease primarily of cattle and dogs, caused by intracellular parasite, Neospora caninum. Neosporosis appears to be a major cause of abortion in dairy cattle worldwide and causes to huge economic loss to dairy industry. Results Recombinant surface associated antigen 1 (NcSAG1, NcSAG1 related sequence 2 (NcSRS2 and the dense granule antigen 2 (NcGRA2 of N. caninum were expressed either in silkworm or in Escherichia coli and purified. The purified recombinant proteins bound to the N. caninum-specific antibodies in serum samples from infected cattle as revealed by an enzyme-linked immunosorbent assay (ELISA. By co-immobilizing these recombinant proteins, a novel indirect ELISA was developed for detection of neosporosis. With the use of 32 serum samples, comprising 12 positive serum samples and 20 negative serum samples, the sensitivity and specificity of the assay were found to be 91.7 and 100%, respectively. Seventy-two serum samples from dairy farms were also tested and one was diagnosed with neosporasis with both this method and a commercial assay. Conclusions A diagnostic method employing recombinant proteins of N. caninum was developed. The method showed high sensitivity and specificity. Diagnostic test with field serum samples suggested its applicability to the practical diagnosis of neosporosis.

  9. A lipemia-independent NanoDrop(®)-based score to identify hemolysis in plasma and serum samples.

    Science.gov (United States)

    Appierto, Valentina; Callari, Maurizio; Cavadini, Elena; Morelli, Daniele; Daidone, Maria Grazia; Tiberio, Paola

    2014-05-01

    The identification and management of hemolyzed samples are crucial issues in the development of new blood-based biomarkers. Using experiments of controlled hemolysis and lipemia and two plasma series from cancer patients, we developed and validated a lipemia-independent hemolysis score (HS). HS resulted strictly associated with the amount of lysed erythrocytes and with serum index measurement (reference method), highly reproducible, and able to identify as hemolyzed plasma/serum samples containing ≥6.1 mg/dl of free hemoglobin. We developed a simple, robust, sensitive, cost-effective, spectrophotometrically-based system to identify hemolyzed plasma/serum specimens. The procedure requires only 2 μl of sample, thus representing a useful tool for research studies and an essential pre-analytical quality control for an optimal biobanking of liquid biopsies.

  10. Immunoaffinity sample purification and MALDI-TOF MS analysis of alpha-Solanine and alpha-chaconine in serum.

    Science.gov (United States)

    Driedger, D R; Sporns, P

    2001-02-01

    A sample purification technique was developed for the detection of potato glycoalkaloids (GAs) in blood serum by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). GAs were extracted from spiked serum (5 mL) using a C(18) solid-phase extraction cartridge. The GAs were then selectively captured on antibody-coated agarose beads. The agarose beads were washed with water and the GAs eluted with 25 microL of methanol. MALDI-TOF MS was used to detect the GAs in the methanol eluent. Immunoaffinity sample purification of the GAs effectively reduced the signal suppression observed during the analysis of unpurified samples. alpha-Chaconine and alpha-solanine were detected in serum spiked with 1 ng/mL of each GA.

  11. Serum cadmium levels in a sample of blood donors in the Western Amazon, Brazil, 2010-2011

    Directory of Open Access Journals (Sweden)

    Andre Ricardo Maia da Costa de Faro

    2014-02-01

    Full Text Available A cross-sectional study was conducted to determine the distribution of serum cadmium (Cd levels in blood donors in Rio Branco, Acre State, Brazil. Blood samples were obtained from 922 volunteer blood donors from 18 to 65 years of age at the Hemoacre blood center in 2010-2011. Mean serum Cd was 0.37µg/L (95%CI: 0.33-0.41. Increased serum Cd was associated with lower schooling; individuals with less than five years of schooling showed a mean Cd of 0.61µg/L (95%CI: 0.34-0.89, compared to 0.34µg/L (95%CI: 0.28-0.40 among those with more than nine years of schooling. Mean serum Cd was three times higher among smokers. Smoking showed a positive association with Cd level, with an OR of 12.36 (95%CI: 7.70-19.84. Meanwhile, serum Cd was lower among individuals that regularly drank tea, as compared to non-tea drinkers. Serum Cd levels were mostly below the reference value (88.3% of participants. Mean serum Cd in the current study indicates that in general the population studied here is not exposed to worrisome Cd levels.

  12. Serum cadmium levels in a sample of blood donors in the Western Amazon, Brazil, 2010-2011.

    Science.gov (United States)

    Faro, André Ricardo Maia da Costa de; Pinto, Wagner de Jesus; Ferreira, Aldo Pacheco; Barbosa Jr, Fernando; Souza, Vanessa Cristina de Oliveira; Fujimoto, Denys Eiti; Koifman, Rosalina Jorge; Koifman, Sérgio

    2014-02-01

    A cross-sectional study was conducted to determine the distribution of serum cadmium (Cd) levels in blood donors in Rio Branco, Acre State, Brazil. Blood samples were obtained from 922 volunteer blood donors from 18 to 65 years of age at the Hemoacre blood center in 2010-2011. Mean serum Cd was 0.37µg/L (95%CI: 0.33-0.41). Increased serum Cd was associated with lower schooling; individuals with less than five years of schooling showed a mean Cd of 0.61µg/L (95%CI: 0.34-0.89), compared to 0.34µg/L (95%CI: 0.28-0.40) among those with more than nine years of schooling. Mean serum Cd was three times higher among smokers. Smoking showed a positive association with Cd level, with an OR of 12.36 (95%CI: 7.70-19.84). Meanwhile, serum Cd was lower among individuals that regularly drank tea, as compared to non-tea drinkers. Serum Cd levels were mostly below the reference value (88.3% of participants). Mean serum Cd in the current study indicates that in general the population studied here is not exposed to worrisome Cd levels.

  13. Survey of Minerai Status of Cattle in the Adamaoua Region of Cameroon

    Directory of Open Access Journals (Sweden)

    Njwe, RM.

    1994-01-01

    Full Text Available Serum and liver samples were collected from Zebu Gudali and Zebu Banyo cattle freshly slaughtered in abattoirs at eight different locations in each of the five administrative divisions of the Adamaoua region of Cameroon during the wet season (September to October, 1983 and the dry season (February to March, 1984. Liver samples were analysed for iron, copper and manganese while serum samples were analysed for calcium, magnesium and copper. Most of the animals were adequate in calcium, magnesium, iron and manganese. Copper was deficient in liver and sera of most of the animals. There is a need to intensif/research on the mineral status of cattle in the Adamaoua region of Cameroon. Copper sulphate is recommended in sait licks for cattle in this region.

  14. Control of declared origin of bovine serum, a pilot study

    Science.gov (United States)

    Horacek, M.; Papesch, W.

    2009-04-01

    Bovine serum is the essential culture medium for cell cultures. Therefore it is highly demanded and the quality of the serum, e.g.: absence of bacteria, viruses certain antibodies, etc.., are important criteria. as some cattle diseases are endemic in certain regions, the origin of bovine serum is an important quality measure for its value. Thus the need to control the declared origins is present. Bovine serum was measured for d2H, d13C, d15N and d34S of proteine (dry residue) and d2H and d18O of the serum water. The hydrogen and oxygen are mainly depending by the isotopic composition of the water ingested by the cattle, and thus usually influenced by the isotopic signal of the precipitation. The carbon isotope signal is reflecting the diet of the cattle, whether it mainly feed on C3- or C4-plants. The nitrogen and sulphur isotope ratio is transferred from the ground/soil into the plant material and into the animal tissue, with some offset for nitrogen and without any significant offset for sulphur. Bovine serum samples from Canada, USA, Mexico, Brazil, Australia and New Zealand have been analysed. Due to the variations in the environmental conditions in different countries and regions which influence the isotope signatures of the serum samples it is possible to discriminate samples of different origin. Main discriminating parameters are d2H and d18O, d13C and d34S.

  15. Bluetongue virus RNA detection by real-time rt-PCR in post-vaccination samples from cattle.

    Science.gov (United States)

    De Leeuw, I; Garigliany, M; Bertels, G; Willems, T; Desmecht, D; De Clercq, K

    2015-04-01

    Bluetongue virus serotype 8 (BTV-8) was responsible for a large outbreak among European ruminant populations in 2006-2009. In spring 2008, a massive vaccination campaign was undertaken, leading to the progressive disappearance of the virus. During surveillance programmes in Western Europe in 2010-2011, a low but significant number of animals were found weakly positive using BTV-specific real-time RT-PCR, raising questions about a possible low level of virus circulation. An interference of the BTV-8 inactivated vaccine on the result of the real-time RT-PCR was also hypothesized. Several studies specifically addressed the potential association between a recent vaccination and BTV-8 RNA detection in the blood of sheep. Results were contradictory and cattles were not investigated. To enlighten this point, a large study was performed to determine the risks of detection of bluetongue vaccine-associated RNA in the blood and spleen of cattle using real-time RT-PCR. Overall, the results presented clearly demonstrate that vaccine viral RNA can reach the blood circulation in sufficient amounts to be detected by real-time RT-PCR in cattle. This BTV-8 vaccine RNA carriage appears as short lasting. © 2013 Blackwell Verlag GmbH.

  16. Microcontroller-based system for estimate of calcium in serum samples.

    Science.gov (United States)

    Neelamegam, Periyaswmy; Jamaludeen, Abdul Sheriff; Ragendran, Annamalai; Murugrananthan, Krishanamoorthy

    2010-01-01

    In this study, a microcontroller-based control unit was designed and constructed for the estimation of serum calcium in blood samples. The proposed optoelectronic instrument used a red light emitting diode (LED) as a light source and photodiode as a sensor. The performance of the system was compared with that of a commercial instrument in measuring calcium ion. The quantitative analysis of calcium in a catalyst using arsenazo III as colorimetric reagent was used to test the device. The calibration curve for calcium binding with arsenazo III was drawn to check the range of linearity, which was between 0.1 to 4.5 mM L⁻¹. The limit of detection (LOD) is 0.05 mM L⁻¹. Absorbance changes over the pH range of 2-12 were determined to optimize the assay, with maximum absorption at pH 9.0. Interferences in absorbance from monovalent (K+ and Na+) and divalent (Mg²+) cations were also studied. The results show that the system works successfully.

  17. A computational platform for MALDI-TOF mass spectrometry data: application to serum and plasma samples.

    Science.gov (United States)

    Mantini, Dante; Petrucci, Francesca; Pieragostino, Damiana; Del Boccio, Piero; Sacchetta, Paolo; Candiano, Giovanni; Ghiggeri, Gian Marco; Lugaresi, Alessandra; Federici, Giorgio; Di Ilio, Carmine; Urbani, Andrea

    2010-01-03

    Mass spectrometry (MS) is becoming the gold standard for biomarker discovery. Several MS-based bioinformatics methods have been proposed for this application, but the divergence of the findings by different research groups on the same MS data suggests that the definition of a reliable method has not been achieved yet. In this work, we propose an integrated software platform, MASCAP, intended for comparative biomarker detection from MALDI-TOF MS data. MASCAP integrates denoising and feature extraction algorithms, which have already shown to provide consistent peaks across mass spectra; furthermore, it relies on statistical analysis and graphical tools to compare the results between groups. The effectiveness in mass spectrum processing is demonstrated using MALDI-TOF data, as well as SELDI-TOF data. The usefulness in detecting potential protein biomarkers is shown comparing MALDI-TOF mass spectra collected from serum and plasma samples belonging to the same clinical population. The analysis approach implemented in MASCAP may simplify biomarker detection, by assisting the recognition of proteomic expression signatures of the disease. A MATLAB implementation of the software and the data used for its validation are available at http://www.unich.it/proteomica/bioinf. (c) 2009 Elsevier B.V. All rights reserved.

  18. Evaluation of HER2/neu oncoprotein in serum & tissue samples of women with breast cancer

    Directory of Open Access Journals (Sweden)

    Shailaja Shukla

    2016-01-01

    Interpretation & conclusions: The results suggest that elevated serum HER2 level was associated with a clinicopathological aggressive phenotype of breast carcinoma and was related to tissue HER2 overexpression. Therefore, serum HER2 may be useful for monitoring the course of the disease and response to treatment.

  19. Prevalence of Antibodies Against Coxiella burnetii in Korean Native Cattle, Dairy Cattle, and Dogs in South Korea.

    Science.gov (United States)

    Lyoo, Kwang-Soo; Kim, Doo; Jang, Hyung Gwan; Lee, Seung-Joon; Park, Mi Yeoun; Hahn, Tae-Wook

    2017-03-01

    Coxiella burnetii is a zoonotic agent and causes coxiellosis, which is a cause of reproductive failure in a range of animal species, including abortion and stillbirth and Q fever, which is most often characterized by an acute flu-like illness, mild pneumonia, and/or hepatitis in humans. While livestock are well recognized worldwide as a source of infection, the zoonotic risk of C. burnetii infection in companion animals such as dogs may be overlooked. For serological diagnosis, indirect immunofluorescent assay (IFA) and enzyme-linked immunosorbent assay (ELISA) are generally considered good methods for prevalence surveys of coxiellosis. In this study, we conducted a nationwide survey of the seroprevalence of previous exposure to C. burnetii in dogs, dairy cattle, and Korean native cattle (a primarily beef breed) in South Korea. Serum samples obtained from 3087 Korean native cattle, 1224 dairy cattle, and 1023 dogs were collected from eight provinces in South Korea, and IFA and ELISA were performed to test for seropositivity. The prevalence of C. burnetii was 1.7% in Korean native cattle, 10.5% in dairy cattle, and 2.9% in dogs. This is the first report identifying previous exposure to C. burnetii in South Korean dogs. Furthermore, the presence of C. burnetii antibodies in companion and feral dogs indicates that dogs can be a potential reservoir species for zoonotic risk of C. burnetii infection in South Korea. Therefore, more detailed studies aiming to clarify epidemiological factors should be performed in the future.

  20. Serum electrolyte values of cows during third trimester of pregnancy ...

    African Journals Online (AJOL)

    Some serum electrolyte values of 170 cows in eight settled cattle herds in Zaria, Northern Nigeria during the third trimester of pregnancy and first two weeks of lactation were determined. The cows were sampled once weekly during the third trimester of pregnancy and early lactation (first two weeks) periods for two ...

  1. Serum biochemical parameters of cows during pregnancy and early ...

    African Journals Online (AJOL)

    user

    ABSTRACT: Some serum electrolyte values of 170 cows in eight settled cattle herds in Zaria, Northern Nigeria during the third trimester of pregnancy and first two weeks of lactation were determined. The cows were sampled once weekly during the third trimester of pregnancy and early lactation (first two weeks) periods for ...

  2. Serum biochemical parameters of cows during pregnancy and early ...

    African Journals Online (AJOL)

    user

    Department of Biochemistry, Ahmadu Bello University, Zaria. ABSTRACT: Some serum electrolyte values of 170 cows in eight settled cattle herds in Zaria, Northern Nigeria during the third trimester of pregnancy and first two weeks of lactation were determined. The cows were sampled once weekly during the third trimester ...

  3. Influence of common preanalytical variations on the metabolic profile of serum samples in biobanks

    Energy Technology Data Exchange (ETDEWEB)

    Fliniaux, Ophelie [University of Picardie Jules Verne, Laboratoire de Phytotechnologie EA 3900-BioPI (France); Gaillard, Gwenaelle [Biobanque de Picardie (France); Lion, Antoine [University of Picardie Jules Verne, Laboratoire de Phytotechnologie EA 3900-BioPI (France); Cailleu, Dominique [Batiment Serres-Transfert, rue de Mai/rue Dallery, Plateforme Analytique (France); Mesnard, Francois, E-mail: francois.mesnard@u-picardie.fr [University of Picardie Jules Verne, Laboratoire de Phytotechnologie EA 3900-BioPI (France); Betsou, Fotini [Integrated Biobank of Luxembourg (Luxembourg)

    2011-12-15

    A blood pre-centrifugation delay of 24 h at room temperature influenced the proton NMR spectroscopic profiles of human serum. A blood pre-centrifugation delay of 24 h at 4 Degree-Sign C did not influence the spectroscopic profile as compared with 4 h delays at either room temperature or 4 Degree-Sign C. Five or ten serum freeze-thaw cycles also influenced the proton NMR spectroscopic profiles. Certain common in vitro preanalytical variations occurring in biobanks may impact the metabolic profile of human serum.

  4. Exploring the concurrent presence of hepatitis A virus genome in serum, stool, saliva, and urine samples of hepatitis A patients.

    Science.gov (United States)

    Joshi, Madhuri S; Bhalla, Shilpa; Kalrao, Vijay R; Dhongade, Ramchandra K; Chitambar, Shobha D

    2014-04-01

    The use of saliva and urine as an alternative to serum samples for detection of anti-hepatitis A virus (HAV) IgM antibodies has been documented. However, these samples remain underreported or unexplored for shedding of HAV. To address this issue, paired serum, stool, saliva, and urine samples collected from hepatitis A patients were screened by reverse transcription polymerase chain reaction for detection of HAV RNA. HAV RNA was detected in 67.6% (44/65), 52.3% (34/65), 8.7% (5/57), and 12.3% (8/65) of the serum, stool, saliva, and urine samples, respectively. Phylogenetic analysis of nucleotide sequences obtained for partial RNA polymerase region grouped HAV strains from all of the clinical samples of the study in subgenotype IIIA. Low frequency of HAV nucleic acid in saliva and urine samples indicates limited utility of these samples in genomic studies on HAV but suggests its potential for transmission and infection of hepatitis A. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. Impact of sample extraction on the accurate measurement of progesterone in human serum by liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Ke, Yuyong; Gonthier, Renaud; Labrie, Fernand

    2017-05-01

    In the present study, the impact of the extraction solvent on the accuracy of endogenous progesterone assay in human serum has been investigated using two selective reaction monitoring (SRM) transitions (315>97 & 315>109). Higher levels of noise and more interference were observed when more polar solvents were used for extraction, thus resulting in serious bias of the measured values of progesterone in serum. This is confirmed by monitoring the ion ratio of 315>97-315>109. This issue could not be easily resolved by changes in MS/MS transitions or chromatography conditions. More bias was observed with the SRM transition 315>109 for the polar solvent extraction. Hexane and 1-chlorobutane (polarity index of 0 and 1, respectively) did provide the cleanest samples with a lower noise level in the chromatograms. Moreover, the measured values of progesterone were not changed with different SRM transitions or longer retention time in search of an improved separation. Recovery tests of progesterone have been performed with 1-chlorobutane in matrices with phosphate buffered saline (PBS) 1x, PBS 1×3% bovine serum albumin (BSA), stripped serum/H2O (1:1) and unstripped serum. The recovery (70%∼80%) consistency is observed not only at different levels but also in different matrices. The equivalent recovery between PBS 1x, PBS 1×3% BSA and unstripped serum shows that the impact of progesterone binding to serum proteins on the measurement accuracy can be avoided with this sample preparation procedure. No significant matrix effect on the determination of progesterone was observed with 1-chlorobutane. Within the range of 12.5-2000pg/mL, a good linearity is observed with R>0.99 and weighting factor 1/X. Bias and covariance efficiency of QCs are within 10%. With 1-chlorobutane as the extraction solvent, the concentration of progesterone was measured where the range for postmenopausal serum is 5.74∼91.7pg/mL, which is well below the reported concentrations of 314 pg/mL∼942pg

  6. Comparison of different methods for extraction and purification of human Papillomavirus (HPV) DNA from serum samples

    Science.gov (United States)

    Azizah, N.; Hashim, U.; Nadzirah, Sh.; Arshad, M. K. Md; Ruslinda, A. R.; Gopinath, Subash C. B.

    2017-03-01

    The affectability and unwavering quality of PCR for indicative and research purposes require effective fair systems of extraction and sanitization of nucleic acids. One of the real impediments of PCR-based tests is the hindrance of the enhancement procedure by substances exhibit in clinical examples. This examination considers distinctive techniques for extraction and cleaning of viral DNA from serum tests in view of recuperation productivity as far as yield of DNA and rate recouped immaculateness of removed DNA, and rate of restraint. The best extraction strategies were the phenol/chloroform strategy and the silica gel extraction methodology for serum tests, individually. Considering DNA immaculateness, extraction technique by utilizing the phenol/chloroform strategy delivered the most tasteful results in serum tests contrasted with the silica gel, separately. The nearness of inhibitors was overcome by all DNA extraction strategies in serum tests, as confirm by semiquantitative PCR enhancement.

  7. Comparison of biuret and refractometry methods for the serum total proteins measurement in ruminants.

    Science.gov (United States)

    Katsoulos, Panagiotis D; Athanasiou, Labrini V; Karatzia, Maria A; Giadinis, Nektarios; Karatzias, Harilaos; Boscos, Constantin; Polizopoulou, Zoe S

    2017-12-01

    Determination of serum total protein concentration is commonly performed by the biuret method. Refractometric measurement is a faster and less expensive alternative but its accuracy has not been determined in ruminants. The purpose of the study was to compare the serum total protein concentrations in cattle, sheep, and goats measured by the biuret method with those obtained by refractometry. Serum total protein concentration was determined in 120 cattle, 67 sheep, and 58 goat blood samples refractometrically and with the biuret method. The data were analyzed with a paired samples t-test, and Passing and Bablok regression equations and Bland and Altman plots were generated. There was a strong linear relationship between the total protein values determined with the refractometer and the biuret method in cattle, sheep, and goats. The statistical accuracy, which represents a bias correction factor that measures the deviation of the best-fit line from the 45° line through the origin, was 90.63% for cattle, 93.05% for sheep, and 91.76% for goats. The mean protein values determined with the refractometer were significantly lower than those measured with the biuret method in cattle and goats (P  .05). The evaluated refractometer was sufficiently accurate for the determination of serum total proteins in cattle, sheep, and goats, although it cannot be used interchangeably with the biuret method. The RIs should be corrected for negative bias based on the created equations. © 2017 American Society for Veterinary Clinical Pathology.

  8. Estimations of cholesterol, triglycerides and fractionation of lipoproteins in serum samples of some Nigerian female subjects

    Directory of Open Access Journals (Sweden)

    E.I. Adeyeye

    2011-04-01

    Full Text Available Blood samples (serum were collected to determine some biochemical parameters: total glycerides (TG, total cholesterol (TC, high density lipoprotein-cholesterol (HDL-C, low density lipoprotein-cholesterol (LDL-C and very low density lipoprotein-cholesterol (VLDL-C in 53 female subjects in Warri, Delta State, Nigeria using the Reflotron® (an auto analyser, supported with the use of questionnaire to get information on age and sex. Age range of the subjects was 18–80 years. The TG levels in all the subjects were < 200 mg/dL; only one subject (1.89% had TC < 200 mg/dL; nine subjects (17.0% had HDL-C ≤ 35 mg/dL; for LDL-C only one subject (1.89% had a desirable level of < 130 mg/dL; for VLDL-C 29 subjects (54.7% had values 17.2 mg/dL and above. For therapeutic decision-making, TC/HDL-C and LDL-C/HDL-C, were calculated. In TC/HDL-C, three subjects (5.66% had values < 4.4 and in LDL-C/HDL-C, 41 subjects (77.4% had values < 4.5. Hence, TC, HDL-C, LDL-C, TC/HDL-C and slightly LDL-C/HDL-C and VLDL-C in the subjects could lead to increase coronary heart diseases. Results were matched for the age and sex of subjects.

  9. Serum Protein Profile Study of Clinical Samples Using High Performance Liquid Chromatography-Laser Induced Fluorescence

    DEFF Research Database (Denmark)

    Karemore, Gopal Raghunath; Ukendt, Sujatha; Rai, Lavanya

    2009-01-01

    The serum protein profiles of normal subjects, patients diagnosed with cervical cancer, and oral cancer were recorded using High Performance Liquid Chromatography combined with Laser Induced Fluorescence detection (HPLC-LIF). Serum protein profiles of the above three classes were tested...... for establishing the ability of HPLC-LIF protein profiling technique for discrimination, using hard clustering and Fuzzy clustering methods. The clustering algorithms have quite successfully classified the profiles as belonging to normal, cancer of cervix, and oral cancer conditions....

  10. Validation of an indirect ELISA to detect antibodies against BoHV-1 in bovine and guinea-pig serum samples using ISO/IEC 17025 standards.

    Science.gov (United States)

    Parreño, Viviana; Romera, S Alejandra; Makek, Lucia; Rodriguez, Daniela; Malacari, Darío; Maidana, Silvina; Compaired, Diego; Combessies, Gustavo; Vena, María Marta; Garaicoechea, Lorena; Wigdorovitz, Andrés; Marangunich, Laura; Fernandez, Fernando

    2010-10-01

    Two ELISAs to quantify antibodies to BoHV-1 in the sera of cattle and immunized guinea pigs were developed and validated using ISO/IEC 17025 standards. The cut-off value of the assay was established at 20% positivity of a high positive control for screening of cattle. Using this threshold, the assay properly classified the OIE bovine reference sera EU1, EU2 and EU3. For vaccine potency testing, a cut-off of 40% was selected for both species. The reliability of the assays, given by their diagnostic sensitivity and specificity, using the threshold of 40% was 89.7% and 100%, respectively, for bovines and 94.9% and 100% for guinea pigs, respectively. There was almost perfect agreement between the ELISA and virus neutralization results. In addition, after vaccination, there was a good correlation between the neutralizing and ELISA antibody titers of the serum from the same bovine or guinea pig, sampled at 60 and 30 days post-vaccination, respectively (R(bovine)=0.88, R(guinea pig)=0.92; p<0.0001). A similar correlation was observed when analyzing the mean antibody titers of groups of vaccinated animals (R(bovine)=0.95 and R(guinea pig)=0.97; p<0.0001), indicating the relevance of the ELISAs for batch to batch vaccine potency testing in the target species and in the laboratory animal model. The intermediate precision of the assays expressed as the relative coefficient of variation (CV) of the positive control assayed over a 3-year period in the same laboratory was 22.2% for bovines and 23.1% for guinea pigs. The reproducibility of both techniques obtained in inter-laboratory assays was CV=12.4% for bovines and CV approximately 0 for guinea pigs, which met the requirements of the OIE (CV<30%). The validated ELISAs represent important methods for vaccine potency testing and for controlling BoHV-1 infections. Copyright (c) 2010 Elsevier B.V. All rights reserved.

  11. Comparison of different mass spectrometric approaches coupled to gas chromatography for the analysis of organochlorine pesticides in serum samples.

    Science.gov (United States)

    Fang, Jing; Wu, Qian; Zhao, Yun; Zhao, Hongzhi; Xu, Shunqing; Cai, Zongwei

    2017-01-01

    Gas chromatography-triple quadrupole mass spectrometry (GC-QqQMS) was applied for the determination of eight organochlorine pesticides (OCPs) in human serum. OCPs were extracted from the serum sample by solid phase extraction (SPE) and analyzed by gas chromatography mass spectrometry (GC-MS) or gas chromatography tandem mass spectrometry (GC-MS/MS). Electron ionization (EI) and negative chemical ionization (NCI) under two data acquisition modes, namely selected ion monitoring (SIM) and multiple reaction monitoring (MRM), were compared. The use of MRM generally provided higher selectivity and sensitivity because less interference from the sample matrix existed. The EI mode is more suitable for less electronegative compounds such as dichlorodiphenyldichloroethanes (DDDs) with detection limits ranging from 0.0060 to 0.060ng/mL. In the NCI mode, MRM analysis provided good and lower detection limits (0.0011-0.0030ng/mL) for pesticides containing more chlorines. The methods were validated by analyzing the pesticides in spiked serum at different levels with recoveries ranged from 83% to 116% and relative standard deviations of less than 10%. The developed method was applied for the determination of the OCPs in real human serum samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. The Prevalence of Brucellosis in Cattle, Goats and Humans in Rural Uganda: A Comparative Study.

    Science.gov (United States)

    Miller, R; Nakavuma, J L; Ssajjakambwe, P; Vudriko, P; Musisi, N; Kaneene, J B

    2016-12-01

    A cross-sectional study was conducted to determine the presence of brucellosis in cattle, goats and humans in farms from south-western Uganda and identify risk factors associated with brucellosis in these three host groups. Data and serum samples were collected from 768 cattle, 315 goats and 236 humans, with 635 samples of bovine milk, from 70 farms in two different study areas in south-western Uganda. Sera from livestock were tested with the Rose Bengal Plate test, using B. abortus and B. melitensis antigens, and human sera were tested with a commercial IgG/IgM lateral flow assay. Milk samples were tested using the OIE-approved milk ring test. Screening tests for brucellosis were positive in 14% of cattle serum, 29% of bovine milk, 17% of goat serum and 11% of human serum samples. There were significant differences in the test prevalence of brucellosis by study site, with levels higher in the study area near Lake Mburo National Park than in the study area near Queen Elizabeth National Park. Multivariable regression models identified risk factors associated with increasing test positivity at the individual and farm levels for cattle, goats and humans. Positive associations were seen between increasing seropositivity of brucellosis in goats, cattle and humans. Results of multivariable analyses suggest that improvements in farm biosecurity and hygiene may reduce the risk of brucellosis on the farm and suggest a role for ticks in bovine brucellosis. Although cattle are the focus of brucellosis control in Uganda, the significant associations between seropositivity in humans and seropositivity in goats suggest that brucellosis in goats may be an important contributor to the epidemiology of the disease on the farm. © 2015 Blackwell Verlag GmbH.

  13. [Comparison and optimization of total ionic strength adjustment buffer during detecting fluoride in trace serum sample by fluoride ion selective electrode method].

    Science.gov (United States)

    Zhou, Zhou; Wang, Hongmei; Zhang, Han; Chen, Yanqing

    2016-03-01

    To probe on the influence by four kinds of total ionic strength regulating buffer (TISAB) in establishing the standard curve and determining the level of fluoride in trace serum sample (rats serum and the calf serum) by fluoride ion selective electrode, and further explore the optimal volume ratio among TISAB and serum samples. Standard curve equations of four TISAB were established to clarify the influence of TISAB kinds. And the trace serum samples (rats and calf) were diluted to 1 times by TISAB, and then levels of fluoride in the samples that added by the standard sodium fluoride were detected, subsequently the influence of TISAB kinds on recovery rate were analyzed. Finally, the optimal ratio among serum sample and each TISAB in different serum was determined according to the recovery rate of the serum sample with different volume ratio. The standard curve of TISAB III sample exhibited best fitting effect (R2 = 0.9926 ), and it also showed the overwhelming superiority with the scope of recovery rates ranged from 90% to 105% in the rat serum standard addition recovery experiments. As calf serum, TISAB IV (recommended by health industry standard) performed superior than others with the scope of recovery rate ranged from 90% to 110% in standard addition recovery experiment. When the volume ratio changed, every kinds of TISAB had an optimal proportion with good recovery rate, only TISAB III with the proportion as 1: 1, nevertheless, those change little affected on the recovery rate in calf serum. The method recommended by health industry standard is only suitable for detecting the trace amounts of fluoride in the serum sample as calf, but not the serum sample as rats.

  14. Heat treatment of serum samples from stray dogs naturally exposed to Dirofilaria immitis and Dirofilaria repens in Romania.

    Science.gov (United States)

    Ciucă, L; Genchi, M; Kramer, L; Mangia, C; Miron, L D; Prete, L Del; Maurelli, M P; Cringoli, G; Rinaldi, L

    2016-07-30

    Pre-heating of serum samples has been shown to reverse false negative antigen tests for Dirofilaria immitis infection in dogs. Here the authors report the results of serum sampling in a population of dogs naturally exposed to D. immitis and Dirofilaria repens infection by testing in ELISA before and after heat treatment. Of 194 dogs sampled from four cities in Romania, D. immitis circulating antigens were found in 16 (8.2%) non heated samples and in 52 (26.8%) heated samples. Of the 108 dogs examined by Knott test, 24 dogs (22.2%) were positive for circulating mf. Subsequent PCR identification showed six dogs had D. immitis mf only, 12 dogs, had only D. repens mf, and 5 were positive for both. Fifty% of dogs with circulating D. immitis mf had positive antigen tests before and after heating, while the other 50% reverted to positive only after heat treatment. Sixty% of dogs with mixed D. immitis/D. repens infection were antigen positive before and after heating, while the other 40% converted to positive after heating. Antigen testing for D. immitis in the 12 dogs with only D. repens mf gave conflicting results. Only two dogs (16%) were antigen negative both before and after heat treatment. Six dogs (50%) became antigen positive after heating and four dogs (30%) were antigen positive both before and after heat treatment. Results would suggest that: false negative result for antigen testing can be reverted by heating of the serum sample; dogs infected with D. repens may have also an occult infection with D. immitis; heat treatment of serum from D. repens-infected dogs can reveal an occult infection with D. immitis. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Seroprevalence of antibody to NcSAG1 antigen of Neospora caninum in cattle from Western Java, Indonesia.

    Science.gov (United States)

    Ichikawa-Seki, Madoka; Guswanto, Azirwan; Allamanda, Puttik; Mariamah, Euis Siti; Wibowo, Putut Eko; Nishikawa, Yoshifumi

    2016-01-01

    Neospora caninum can cause fetal abortion and neonatal mortality in cattle, and is a cause of economic concern worldwide. This study aimed to determine the prevalence of Neospora caninum-specific antibodies in cattle from Western Java, Indonesia. Serum samples from 991 cattle from 21 locations were tested for antibodies to N. caninum by using an enzyme-linked immunosorbent assay (ELISA) on the basis of recombinant NcSAG1. The overall seroprevalence was 16.6%, ranging from 0 to 87.5% in the sampled locations. The results of this study indicate latent infection rates of sampled animals were different in each location. Further studies are necessary to elucidate the relationship between N. caninum infection and abortion in cattle, and to identify risk factors for infection in high-prevalence environments.

  16. Variation in Bluetongue virus real-time reverse transcription polymerase chain reaction assay results in blood samples of sheep, cattle, and alpaca.

    Science.gov (United States)

    Brito, Barbara P; Gardner, Ian A; Hietala, Sharon K; Crossley, Beate M

    2011-07-01

    Bluetongue is a vector-borne viral disease that affects domestic and wild ruminants. The epidemiology of this disease has recently changed, with occurrence in new geographic areas. Various real-time quantitative reverse transcription polymerase chain reaction (real-time qRT-PCR) assays are used to detect Bluetongue virus (BTV); however, the impact of biologic differences between New World camelids and domestic ruminant samples on PCR efficiency, for which the BTV real-time qRT-PCR was initially validated are unknown. New world camelids are known to have important biologic differences in whole blood composition, including hemoglobin concentration, which can alter PCR performance. In the present study, sheep, cattle, and alpaca blood were spiked with BTV serotypes 10, 11, 13, and 17 and analyzed in 10-fold dilutions by real-time qRT-PCR to determine if species affected nucleic acid recovery and assay performance. A separate experiment was performed using spiked alpaca blood subsequently diluted in 10-fold series in sheep blood to assess the influence of alpaca blood on performance efficiency of the BTV real-time qRT-PCR assay. Results showed that BTV-specific nucleic acid detection from alpaca blood was consistently 1-2 logs lower than from sheep and cattle blood, and results were similar for each of the 4 BTV serotypes analyzed.

  17. Comparison of KRAS Mutation Assessment in Tumor DNA and Circulating Free DNA in Plasma and Serum Samples

    Directory of Open Access Journals (Sweden)

    Shethah R. Morgan

    2012-01-01

    Full Text Available Testing for mutations in the KRAS oncogene for patients with metastatic colorectal cancer (mCRC is generally performed using DNA from formalin-fixed paraffin-embedded tumor tissue; however, access to specimens can be limited and analysis challenging. This study assessed the identification of KRAS mutations in circulating free DNA (cfDNA using a commercially available KRAS polymerase chain reaction (PCR kit. Matched plasma, serum and tumor samples were available from 71 patients with mCRC who had received prior therapy but whose disease progressed following therapy. Yields of cfDNA from plasma and serum samples were comparable. Analyses were successful in 70/71 plasma-extracted samples (specificity: 97%, sensitivity: 31% and 67/71 serum-extracted samples (specificity: 100%, sensitivity: 25%. This study demonstrates that KRAS mutations can be detected in cfDNA using a commercially available KRAS PCR kit, confirming cfDNA as a potential alternative source of tumor DNA in a diagnostic setting if access to archival tumor specimens is limited.

  18. Pre-Analytical Sample Quality : Metabolite Ratios as an Intrinsic Marker for Prolonged Room Temperature Exposure of Serum Samples

    NARCIS (Netherlands)

    Anton, Gabriele; Wilson, Rory; Yu, Zhong-hao; Prehn, Cornelia; Zukunft, Sven; Adamski, Jerzy; Heier, Margit; Meisinger, Christa; Roemisch-Margl, Werner; Wang-Sattler, Rui; Hveem, Kristian; Wolffenbuttel, Bruce; Peters, Annette; Kastenmueller, Gabi; Waldenberger, Melanie

    2015-01-01

    Advances in the "omics" field bring about the need for a high number of good quality samples. Many omics studies take advantage of biobanked samples to meet this need. Most of the laboratory errors occur in the pre-analytical phase. Therefore evidence-based standard operating procedures for the

  19. Acute phase proteins in cattle after exposure to complex stress

    DEFF Research Database (Denmark)

    Lomborg, S. R.; Nielsen, L. R.; Heegaard, Peter M. H.

    2008-01-01

    Abstract Stressors such as weaning, mixing and transportation have been shown to lead to increased blood concentrations of acute phase proteins (APP), including serum amyloid A (SAA) and haptoglobin, in calves. This study was therefore undertaken to assess whether SAA and haptoglobin levels...... concentrations of SAA and haptoglobin increased significantly in response to the stressors (P...... in blood mirror stress in adult cattle. Six clinically healthy Holstein cows and two Holstein heifers were transported for four to six hours to a research facility, where each animal was housed in solitary tie stalls. Blood samples for evaluation of leukocyte counts and serum SAA and haptoglobin...

  20. Influence of vitamin D on fecal shedding of Escherichia coli O157:H7 in naturally colonized cattle.

    Science.gov (United States)

    Edrington, Tom S; Farrow, Russell L; Mackinnon, Kathryn M; Callaway, Todd R; Anderson, Robin C; Nisbet, David J

    2012-02-01

    Three experiments were conducted to evaluate the influence of vitamin D on fecal shedding of Escherichia coli O157:H7 in cattle. In the first experiment, two groups of cattle (beef and dairy) were assigned to a control treatment or to receive 0.5 × 10(6) IU vitamin D per day via oral bolus for 10 days. Fecal samples were collected before and throughout the dosing period for culture of E. coli O157:H7. No differences were observed for fecal shedding of E. coli O157:H7 among treatments for either beef or dairy animals. Serum concentrations of vitamin D were markedly higher (P vitamin D dosages (2,400, 4,800, and 9,600 IU/day; 14 days each) were administered to 14 dairy steers (7 steers served as controls), fecal samples were collected daily, and serum samples were collected weekly throughout the 42-day experimental period. No significant differences in fecal prevalence or serum vitamin D concentrations were observed for any of the vitamin D dosages. A third experiment sampled feedlot cattle (winter and summer) to determine whether serum vitamin D concentrations were correlated with fecal shedding of E. coli O157:H7. A fecal sample and a blood sample were obtained in each season from 60 randomly selected animals (total of 120 fecal samples and 120 corresponding blood samples). As expected, season was highly correlated (r = 0.66) with serum vitamin D concentration with higher concentrations (P coli O157:H7 prevalence (percentage of positive samples) was not highly correlated (r = 0.16) with season, although the correlation tended to be significant (P = 0.08). The proportion of cattle shedding E. coli O157:H7 was 16.7 and 6.7% for the summer and winter collections, respectively. Results of this research do not support a correlation between vitamin D intake and E. coli O157:H7 shedding in cattle.

  1. Comparison of GC-MS and GC×GC-MS in the analysis of human serum samples for biomarker discovery.

    Science.gov (United States)

    Winnike, Jason H; Wei, Xiaoli; Knagge, Kevin J; Colman, Steven D; Gregory, Simon G; Zhang, Xiang

    2015-04-03

    We compared the performance of gas chromatography time-of-flight mass spectrometry (GC-MS) and comprehensive two-dimensional gas chromatography mass spectrometry (GC×GC-MS) for metabolite biomarker discovery. Metabolite extracts from 109 human serum samples were analyzed on both platforms with a pooled serum sample analyzed after every 9 biological samples for the purpose of quality control (QC). The experimental data derived from the pooled QC samples showed that the GC×GC-MS platform detected about three times as many peaks as the GC-MS platform at a signal-to-noise ratio SNR ≥ 50, and three times the number of metabolites were identified by mass spectrum matching with a spectral similarity score Rsim ≥ 600. Twenty-three metabolites had statistically significant abundance changes between the patient samples and the control samples in the GC-MS data set while 34 metabolites in the GC×GC-MS data set showed statistically significant differences. Among these two groups of metabolite biomarkers, nine metabolites were detected in both the GC-MS and GC×GC-MS data sets with the same direction and similar magnitude of abundance changes between the control and patient sample groups. Manual verification indicated that the difference in the number of the biomarkers discovered using these two platforms was mainly due to the limited resolution of chromatographic peaks by the GC-MS platform, which can result in severe peak overlap making subsequent spectrum deconvolution for metabolite identification and quantification difficult.

  2. Cytokine and hormonal profile in serum samples of patients undergoing controlled ovarian stimulation: interleukin-1beta predicts ongoing pregnancy.

    Science.gov (United States)

    Bonetti, T C S; Salomao, R; Brunialti, M; Braga, D P A F; Borges, E; Silva, I D C G

    2010-08-01

    Changes in the endometrium are not regulated exclusively by ovarian hormones; the immune system has also been implicated in normal endometrial function, similar to processes taking place during inflammatory and reparative path. Many cytokines are crucially important for reproductive processes, and the role of cytokines in the female reproductive system function has been broadly investigated during controlled ovarian stimulation (COS) for IVF attempts. The aim of this study was to evaluate the levels of serum cytokines and hormones, and the clinical outcomes of women who underwent COS and ICSI procedures. The study prospectively included 96 patients (aged 22-43 years, unexplained or male infertility, n = 61; female infertility factors, n = 35) who underwent ICSI cycles. Serum levels of interleukin (IL-8, IL-6, IL-1beta, IL-10, IL-12), tumour necrosis factor and leukaemia-inhibitory factor (LIF) and the hormones FSH, estradiol, progesterone, anti-Mullerian hormone and Inhibin-B were measured on the day of oocyte retrieval. The ongoing pregnancy rate was 25.3%. The presence of serum IL-1beta positively affected the implantation rate (P = 0.004) and increased the chance of becoming pregnant by 15 fold. Furthermore, the percentage of patients with detectable serum IL-1beta levels who conceived (62.5%) was higher than those who failed to conceive (37.5%; P = 0.019). The LIF was undetectable in all serum samples, and no other factors influenced the clinical outcomes of patients undergoing ICSI cycles. Our findings revealed that detectable serum levels of IL-1beta on the day of oocyte retrieval in patients undergoing COS and ICSI are predictive of successful implantation and ongoing pregnancy.

  3. Sero-prevalence of Schistosoma species in cattle in Maiduguri Metropolis and Jere Local Government Areas of Borno State, Nigeria

    Directory of Open Access Journals (Sweden)

    Idris Umar Hambali

    2016-03-01

    Full Text Available Objectives: This study was designed to investigate the Sero-prevalence of Schistosoma species in cattle in Maiduguri Metropolis (MMC and Jere Local Government Areas (LGAs of Borno State, Nigeria. Materials and Method: Blood samples (n=200 from cattle were collected using a multistage sampling technique; 100 samples each were collected from MMC and Jere LGAs, respectively. The samples were subjected to screening for Schistosoma antibodies using Enzyme Linked Immunosorbent Assay (ELISA. Age, sex, breed and location of cattle were recorded. Results: The overall prevalence of Schistosoma infection among cattle in MMC and Jere LGAs was 10%. Jere LGA had a prevalence rate of 14% and MMC had 6%. At the ward levels, Custom Area in Jere LGA had the highest number of Schistosoma positive (50%. Out of 103 female and 97 male cattle screened, the prevalence of Schistosoma infection in female and male were 9.71% (n=10/103 and 10.31% (n=10/103. Out of the 177 serum samples from cattle aging >1-year (adult examined, 16 (9.04% were positive, while only 4 (17.39% out of 23 serum samples from cattle aging <1-year (young were positive. Out of the eight (8 breeds screened, the highest number of cases was recorded in Kuri breed (16.22%. This was followed by Sokoto Gudali (10.9% breed. The prevalence in other breeds was as follows: Abore- 10%, Red Bororo- 5.26%, and White Fulani- 6.67%. Conclusion: It is concluded that schistosomiasis in cattle was prevalent in MMC and Jere LGAs of Borno State. A regular checking program is suggested to constantly check out whether the prevalence rate is increasing, so that effective control measures can be strenthened. [J Adv Vet Anim Res 2016; 3(1.000: 56-61

  4. Investigation of dual-layer membrane cloaking method by surface plasmon resonance for direct chronoamperometric immunoassay of serum sample.

    Science.gov (United States)

    Dai, Zong; Yang, Yan; Wu, Hai; Zou, Xiao-Yong

    2011-10-15

    A "dual-layer membrane cloaking" (DLMC) method was developed to construct disposable electrochemical immunosensor for direct determination of serum sample. Mouse IgG (MIgG) molecules were firstly immobilized on a substrate. After the formation of a didodecyldimethylammonium bromide (DDAB) membrane on the MIgG modified substrate, an additional bovine serum albumin (BSA) thin layer was formed to build a BSA/DDAB dual-layer membrane (DLM). When alkaline phosphatase conjugated anti-mouse IgG antibodies (anti-MIgG-ALP) in human serum were incubated on the substrate, anti-MIgG-ALP was recognized specifically by the immobilized MIgG while all nonspecifically adsorbed proteins were selectively removed together with BSA/DDAB DLM by 5% Triton X-100 (v/v) before final measurements. The BSA/DDAB DLM was characterized and optimized by surface plasmon resonance (SPR) technique, and further employed in a disposable immunoassay based on an ITO chip. Under optimal conditions, MIgG in human serum was directly detected in the range of 2.0-18.0 ng mL(-1) without dilution or separation. A limit of detection as low as 0.922 ng mL(-1) (6.15 pM) was obtained. The proposed DLMC method can efficiently prevent the penetration of matrix proteins through single cloaking membrane and completely eliminate nonspecific adsorption. It has great potential in providing a versatile way for direct determination of serum sample with ultra-sensitivity. Copyright © 2011 Elsevier B.V. All rights reserved.

  5. Leptospirosis in Cattle From Markets of Almaty Province, Kazakhstan

    Directory of Open Access Journals (Sweden)

    Kirkimbayeva Zhumagul

    2015-04-01

    Full Text Available This paper is the first study of the prevalence of leptospirosis in the cattle at slaughter from a rural area of Kazakhstan. Five hundred and seventy three samples of serum, urine, and kidneys from cattle of Alatau, Kazakh white and Auliyekol breed, aged from 2 to 5 years (unknown vaccination status, from the province of Almaty in the South-Eastern region were collected during four years (March 2010 to October 2013. The serological, bacteriological, and molecular analyses were performed. Serum samples were tested with 14 reference Leptospira serovars by microscopic agglutination test (MAT. MAT results showed that 89 (15.53% serum samples had detectable antibodies against seven serovars of L. interrogans at a dilution of ≥1:100. Serovars: Pomona (38.2%, Tarassovi (27.2%, and Kabula (18.8% were the most prevalent and their titres ranged from 100 to 1200. The spirochetes were detected in 11 samples of urine and nine samples of kidneys under dark-field microscope observation. The pure cultures were obtained from three samples. PCR technique confirmed leptospirosis in 23 out of 89 urine samples from cows, which showed the presence of leptospiral antibodies in microagglutination test. The high disease prevalence in cows indicates the high Leptospira contamination in this area. It was concluded that the bovine leptospirosis is an endemic and locally widespread disease in Kazakhstan, and that it may play a role in zoonotic transmission to humans.

  6. Flow injection analysis-Rayleigh light scattering detection for online determination of protein in human serum sample.

    Science.gov (United States)

    Li, Ying; Dong, Lijun; Wang, Weiping; Hu, Zhide; Chen, Xingguo

    2006-07-01

    A flow injection analysis (FIA) system combined with Rayleigh light scattering (RLS) detection is developed for the sensitive and rapid determination of protein concentration in human serum sample. This method is based on the weak intensity of RLS of Eriochrome Black T (EBT, 2-hydroxy-1-(1-hydroxy-2-naphthylazo)-6-nitronaphthalene-4-sulfonic acid sodium salt), which can be enhanced by the addition of protein in weakly acidic solution. The effects of pH and interfering species on the determination of protein were examined. Calibrations for protein, based on RLS intensity, were linear in the concentration ranges of 7-36 microg/ml for human serum album (HSA) and 8-44 microg/ml for bovine serum album (BSA). The detection limits of the method were found to be 0.882 and 2.507 microg/ml for HSA and BSA, respectively. A relative standard deviation of 0.76% (n=5) was obtained with 20 microg/ml HSA standard solution. The FIA-RLS method was more stable than the general RLS method, and the average RSD value of FIA-RLS was less than that of the general RLS. The sample rate was determined to be 90 samples per hour.

  7. Detection of Wuchereria bancrofti DNA in paired serum and urine samples using polymerase chain reaction-based systems.

    Science.gov (United States)

    Ximenes, Camila; Brandão, Eduardo; Oliveira, Paula; Rocha, Abraham; Rego, Tamisa; Medeiros, Rafael; Aguiar-Santos, Ana; Ferraz, João; Reis, Christian; Araujo, Paulo; Carvalho, Luiz; Melo, Fabio L

    2014-12-01

    The Global Program for the Elimination of Lymphatic Filariasis (GPELF) aims to eliminate this disease by the year 2020. However, the development of more specific and sensitive tests is important for the success of the GPELF. The present study aimed to standardise polymerase chain reaction (PCR)-based systems for the diagnosis of filariasis in serum and urine. Twenty paired biological urine and serum samples from individuals already known to be positive for Wuchereria bancrofti were collected during the day. Conventional PCR and semi-nested PCR assays were optimised. The detection limit of the technique for purified W. bancrofti DNA extracted from adult worms was 10 fg for the internal systems (WbF/Wb2) and 0.1 fg by using semi-nested PCR. The specificity of the primers was confirmed experimentally by amplification of 1 ng of purified genomic DNA from other species of parasites. Evaluation of the paired urine and serum samples by the semi-nested PCR technique indicated only two of the 20 tested individuals were positive, whereas the simple internal PCR system (WbF/Wb2), which has highly promising performance, revealed that all the patients were positive using both samples. This study successfully demonstrated the possibility of using the PCR technique on urine for the diagnosis of W. bancrofti infection.

  8. Detection of Wuchereria bancrofti DNA in paired serum and urine samples using polymerase chain reaction-based systems

    Directory of Open Access Journals (Sweden)

    Camila Ximenes

    2014-12-01

    Full Text Available The Global Program for the Elimination of Lymphatic Filariasis (GPELF aims to eliminate this disease by the year 2020. However, the development of more specific and sensitive tests is important for the success of the GPELF. The present study aimed to standardise polymerase chain reaction (PCR-based systems for the diagnosis of filariasis in serum and urine. Twenty paired biological urine and serum samples from individuals already known to be positive for Wuchereria bancrofti were collected during the day. Conventional PCR and semi-nested PCR assays were optimised. The detection limit of the technique for purified W. bancrofti DNA extracted from adult worms was 10 fg for the internal systems (WbF/Wb2 and 0.1 fg by using semi-nested PCR. The specificity of the primers was confirmed experimentally by amplification of 1 ng of purified genomic DNA from other species of parasites. Evaluation of the paired urine and serum samples by the semi-nested PCR technique indicated only two of the 20 tested individuals were positive, whereas the simple internal PCR system (WbF/Wb2, which has highly promising performance, revealed that all the patients were positive using both samples. This study successfully demonstrated the possibility of using the PCR technique on urine for the diagnosis of W. bancrofti infection.

  9. Restricted access carbon nanotubes for direct extraction of cadmium from human serum samples followed by atomic absorption spectrometry analysis.

    Science.gov (United States)

    Barbosa, Adriano F; Barbosa, Valéria M P; Bettini, Jefferson; Luccas, Pedro O; Figueiredo, Eduardo C

    2015-01-01

    In this paper, we propose a new sorbent that is able to extract metal ions directly from untreated biological fluids, simultaneously excluding all proteins from these samples. The sorbent was obtained through the modification of carbon nanotubes (CNTs) with an external bovine serum albumin (BSA) layer, resulting in restricted access carbon nanotubes (RACNTs). The BSA layer was fixed through the interconnection between the amine groups of the BSA using glutaraldehyde as cross-linker. When a protein sample is percolated through a cartridge containing RACNTs and the sample pH is higher than the isoelectric point of the proteins, both proteins from the sample and the BSA layer are negatively ionized. Thus, an electrostatic repulsion prevents the interaction between the proteins from the sample on the RACNTs surface. At the same time, metal ions are adsorbed in the CNTs (core) after their passage through the chains of proteins. The Cd(2+) ion was selected for a proof-of-principle case to test the suitability of the RACNTs due to its toxicological relevance. RACNTs were able to extract Cd(2+) and exclude almost 100% of the proteins from the human serum samples in an online solid-phase extraction system coupled with thermospray flame furnace atomic absorption spectrometry. The limits of detection and quantification were 0.24 and 0.80 μg L(-1), respectively. The sampling frequency was 8.6h(-1), and the intra- and inter-day precisions at the 0.80, 15.0, and 30.0 μg L(-1) Cd(2+) levels were all lower than 10.1% (RSD). The recoveries obtained for human blood serum samples fortified with Cd(2+) ranged from 85.0% to 112.0%. The method was successfully applied to analyze Cd(2+) directly from six human blood serum samples without any pretreatment, and the observed concentrations ranged from

  10. A simple and sensitive method for lipoprotein and lipids profiles analysis of individual micro-liter scale serum samples.

    Science.gov (United States)

    Yang, Liu; Fan, Baoyan; Yang, Kangmin; Zhu, Haibo

    2012-02-01

    A simple and sensitive method to determine lipoprotein and lipids profiles in micro-liter scale individual serum sample is not presently available. Traditional lipoprotein separation techniques either by ultra-centrifugation or by liquid chromatography methods have their disadvantages in both lipoprotein separation and lipids component quantification. In this study we used small volume needing size-exclusion fast protein liquid chromatography to separate different lipoprotein subclasses in 50μL serum. And lipids contents, such as cholesterol, cholesterol ester and triacylglycerol, were measured by using two different fluorescence-based lipid detection methods. With this method, very low density lipoprotein, low density lipoprotein and high density lipoprotein could be easily separated, and follow-up lipid detection was completed by simple kinds of reactions. Serum lipoprotein and lipids profiling from C57BL/6 mice (n=5) and human (n=5) were analyzed. The elution profiles of five individuals were highly reproducible, and there were lipoprotein and lipids distribution variations between C57BL/6 mice and human beings. In conclusion, this method which combined small volume needing size-exclusion fast protein liquid chromatography and fluorescence-based lipids measurement, provided a simple, efficient, integrity and reproducible procedure for determining serum lipoprotein and lipids profiles in micro-liter scale levels. It becomes possible that determination of lipoprotein profiles and gaining information of lipids in different lipoproteins can be accomplished simultaneously. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  11. Brucellosis in adult beef cattle of Mexican origin shipped direct-to-slaughter into Texas.

    Science.gov (United States)

    Brown, W H; Hernández de Anda, J

    1998-03-01

    To compare prevalence estimates of brucellosis (BR) in adult beef cattle that originated from different states and regions of Mexico and that were shipped direct-to-slaughter into Texas during 1995. Epidemiologic survey. About 65,000 adult beef cattle. Blood samples were collected during postmortem examinations and were tested for serum antibodies to Brucella abortus, using the particle concentration fluorescence immunoassay and automated complement-fixation test. Prevalence estimates and 95% confidence intervals of BR were calculated by state of origin in Mexico. The difference among prevalence estimates of BR in cattle from different states and regions was tested for significance (P Aguascalientes (7.89%), and Campeche (12.24%). In addition, prevalence estimates of BR in cattle were significantly different among the northern (0.22%), south-central (3.18%), and south coastal (9.42%) regions of Mexico. Results of this study indicate that the number of cattle exposed to B abortus may be significantly different among states and regions of Mexico. Current import sanitary requirements should continue to mitigate potential risk of transmission of BR from sexually intact cattle of Mexican origin to Texas cattle.

  12. A comparative study of some physico-chemical properties of human serum albumin samples from different sources--I : Some physico-chemical properties of isoionic human serum albumin solutions

    NARCIS (Netherlands)

    Dröge, J.H.M.; Janssen, L.H.M.; Wilting, J.

    1982-01-01

    Human serum albumin samples from different sources were investigated. The fatty acid content of the albumin before and after deionization on a mixed bed ion-exchange column varied from sample to sample. When an albumin sample from one source was deionized under standard conditions the amount of

  13. Characterization of three commercial ELISA kits for detection of BOHV-1 gE specific antibodies in serum and milk samples and applicability of bulk milk for determination of herd status.

    Science.gov (United States)

    Tignon, Marylène; De Baere, Miet; Hanon, Jean-Baptiste; Goolaerts, Annelies; Houtain, Jean-Yves; Delooz, Laurent; Cay, Ann Brigitte

    2017-07-01

    Vaccination of animals with gE-deleted vaccine strains (gE- marker vaccines) and differential detection of vaccinated vs infected animals with antibody ELISA targeting the gE or the gB proteins have been proved to be useful tools in programs for control and eradication of the bovine herpesvirus 1 (BoHV-1) responsible for infectious bovine rhinotracheitis (IBR), a major pathogen of cattle. The diagnostic sensitivity (DSe) and specificity (DSp) of three commercial gE ELISA kits from IDEXX, IDVet and CIV-HIPRA were compared for serum and milk matrices. Limiting the analysis to 198 individual with concordant ELISA results in serum (91 naïve, 37 vaccinated and 70 infected) the DSe of gE kits was estimated to 0,97 for IDEXX, 0,93 for CIV-HIPRA and 0,53 for IDVet using milk samples and the DSp to 0,95 for IDEXX, 1,00 for IDVet and CIV-HIPRA. The applicability of gE ELISA for individual or bulk milk testing as an additional tool in control programs dedicated to the certification and control of vaccinated herds was evaluated. Two of the three evaluated gE ELISA kits presented substantial to good agreement individual milk and serum samples. The bulk-tank milk also proved to be suitable for the detection of BoHV-1 in vaccinated herds provided that gE prevalence is superior to 10% as false negative results are often observed at lower gE herd prevalence. This limitation could be reduced to 8% of prevalence when a prior concentration step was applied to bulk milk samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Development of combination tapered fiber-optic biosensor dip probe for quantitative estimation of interleukin-6 in serum samples

    Science.gov (United States)

    Wang, Chun Wei; Manne, Upender; Reddy, Vishnu B.; Oelschlager, Denise K.; Katkoori, Venkat R.; Grizzle, William E.; Kapoor, Rakesh

    2010-11-01

    A combination tapered fiber-optic biosensor (CTFOB) dip probe for rapid and cost-effective quantification of proteins in serum samples has been developed. This device relies on diode laser excitation and a charged-coupled device spectrometer and functions on a technique of sandwich immunoassay. As a proof of principle, this technique was applied in a quantitative estimation of interleukin IL-6. The probes detected IL-6 at picomolar levels in serum samples obtained from a patient with lupus, an autoimmune disease, and a patient with lymphoma. The estimated concentration of IL-6 in the lupus sample was 5.9 +/- 0.6 pM, and in the lymphoma sample, it was below the detection limit. These concentrations were verified by a procedure involving bead-based xMAP technology. A similar trend in the concentrations was observed. The specificity of the CTFOB dip probes was assessed by analysis with receiver operating characteristics. This analysis suggests that the dip probes can detect 5-pM or higher concentration of IL-6 in these samples with specificities of 100%. The results provide information for guiding further studies in the utilization of these probes to quantify other analytes in body fluids with high specificity and sensitivity.

  15. Relationship between clinical parameters and cytokine profiles in inflamed gingival tissue and serum samples from patients with chronic periodontitis.

    Science.gov (United States)

    Górska, Renata; Gregorek, Hanna; Kowalski, Jan; Laskus-Perendyk, Agnieszka; Syczewska, Małgorzata; Madaliński, Kazimierz

    2003-12-01

    The purpose of the present study was to assess the relation between clinical parameters and concentrations of the key (IL-1beta, TNF-alpha, IL-2, IFN-gamma, IL-4, IL-10) cytokines, important in the initiation and progression of periodontal diseases, within inflamed gingival tissues and serum samples from patients with severe chronic periodontitis. Twenty-five patients with severe chronic periodontitis, who had sites with probing depths (PD) > 5 mm, and 25 periodontally healthy persons were included in the study. Clinical examinations including PD, clinical attachment loss, plaque index, and bleeding index were performed before periodontal treatment. Gingival tissue biopsies were collected from one active site of each patient and from healthy individuals, and blood samples were withdrawn on the day of tissue biopsy. The concentrations of cytokines were determined by an enzyme-linked immunosorbent assay, and the relationship between their profiles in situ and in circulation with clinical parameters was analysed. The concentrations of IL-1beta, TNF-alpha, IL-2, IFN-gamma were, on average, significantly higher in serum samples and gingival tissue biopsies from periodontitis patients than in healthy controls. However, serum samples from both groups showed high individual variability of cytokine profiles, and no association between cytokine concentrations and clinical parameters of periodontitis was found. On the contrary, the levels of IL-4 and IL-10 in both kinds of samples obtained from patients and controls were generally low or even undetectable, and remained, on average, on the same level. However, the frequency of IL-4 (88% positive samples) and IL-10 (72%) was much higher in healthy gingival tissues. High concentrations of TNF-alpha, IFN-gamma and IL-2 and, especially, a high ratio of IL-1beta/IL-10 and TNF-alpha/IL-4 found in tissue biopsies from periodontitis patients, strongly correlated with the severity of periodontitis. These results indicate that high

  16. Prevalence of Antibodies Against Foot-and-Mouth Disease Virus in Cattle in Kasese and Bushenyi Districts in Uganda

    DEFF Research Database (Denmark)

    Mwiine, F. N.; Ayebazibwe, C.; Olaho-Mukani, W.

    2010-01-01

    Abstract: The aim of this study was to determine the seroprevalence and serotype-specificity of the circulating antibodies against Foot-and-Mouth Disease Virus (FMDV) in cattle in K asese and Bushenyi districts in Uganda. A total of 309 serum samples were collected and tested for antibodies against...

  17. Characterization of implant materials in fetal bovine serum and sodium sulfate by electrochemical impedance spectroscopy. II. Coarsely sandblasted samples.

    Science.gov (United States)

    Contu, F; Elsener, B; Böhni, H

    2003-10-01

    Electrochemical impedance spectroscopy is used to investigate the corrosion resistance of coarsely sandblasted implant alloys, commercially pure titanium, Ti6Al4V, Ti6Al7Nb, and CoCrMo in 0.1M sodium sulfate and fetal bovine serum. Coarsely sandblasted samples have a heterogeneous surface constituted by a large number of protrusions and recessions. Impedance spectra collected in sodium sulfate present two time constants (maxima in the phase-angle of the bode plot) associated with the total surface and with the tips, respectively. In bovine serum, the two maxima in the impedance spectra cannot be distinguished because of the formation of an adsorption layer of organic molecules, which causes a decrease in the values of both the total and tips' capacitances as well as an increase in the polarization resistance. Ti6Al4V and Ti6Al7Nb show the highest corrosion rate both in serum and in sodium sulfate. Based on the capacitance values obtained in sodium sulfate, the real surface area of the coarsely sandblasted electrodes has been estimated relative to mechanically polished surfaces. The values of the effective electrode area correlate with the mechanical properties of the samples: in fact, the softest electrode (commercially pure titanium) shows the largest effective electrode area, whereas the hardest material (CoCrMo alloy) shows the smallest surface area. Copyright 2003 Wiley Periodicals, Inc. J Biomed Mater Res 67A: 246-254, 2003

  18. Distribution and correlates of serum 25-hydroxyvitamin D levels in a sample of patients with hip fracture.

    Science.gov (United States)

    Pieper, Carl F; Colon-Emeric, Cathleen; Caminis, John; Betchyk, Kathleen; Zhang, Jie; Janning, Cheri; Shostak, John; LeBoff, Meryl S; Heaney, Robert R; Lyles, Kenneth W

    2007-12-01

    Vitamin D deficiency is common in older populations, particularly during the winter months due to low levels of ultraviolet light exposure, and in nursing home residents. The main objective of the current study was to assess the distribution of serum 25-hydroxyvitamin D and its correlates in a sample of men and women with recent hip fractures who were part of a large clinical trial. This was a cross-sectional exploratory study of screened and ultimately randomized patients with hip fractures. They were part of a multinational (115 clinical centers in 20 countries), randomized, placebo-controlled, double-blind study testing the efficacy of a yearly IV bisphosphonate (zoledronic acid) in the prevention of new clinical fractures in patients with recent hip fracture repair. Levels of 25-hydroxyvitamin D, calcium, alkaline phosphatase, creatinine clearance, and albumin were measured at a screening visit using blood serum. Demographic variables were assessed by patient self-report. Bone mineral density (BMD) was assessed by dual-energy x-ray absorptiometry of the nonfractured hip. This report included 1174 screened patients (526 with vitamin D measured) and 655 (385 with vitamin D measured) patients randomized before the protocol amendment. In screened patients, levels of 25-hydroxyvitamin D were low (median, 14.7 ng/mL; interquartilc range, 7.6, 21.65). Overall, 51% were at or below the clinically meaningful threshold of 15 ng/mL. Among those patients randomized, the level of 25-hydroxyvitamin D was significantly positively related to male sex (rho, 0.13; P femoral neck (rho, 0.22; P fracture. This insufficiency was related only to serum calcium in multivariable controlled models but cannot be reliably identified or excluded by measuring serum calcium alone. Physicians should be encouraged to check and monitor patients' serum levels of 25-hydroxyvitamin D.

  19. Amperometric immunosensor for the determination of IgA deficiency in human serum samples

    OpenAIRE

    Carlos Rosales-Rivera, Luis; Lluis Acero-Sanchez, Josep; Lozano-Sanchez, Pablo; Katakis, Ioanis; O'Sullivan, Ciara K.

    2012-01-01

    10.1016/j.bios.2011.12.040 An electrochemical immunosensor for the detection of human IgA deficiency in real human blood serum has been developed. The performance of the immunosensor presents a large but sensitive dynamic range that allows the determination of non-deficient IgA levels (>70 µg/mL) as well as of severe IgA deficiencies (0.5¿5.0 µg/mL). The assay architecture involves the immobilisation of a coating antibody on an electrode surface using carboxylic-ended bipodal alkan...

  20. Detection of foot-and-mouth disease virus in the breath of infected cattle using a hand-held device to collect aerosols

    DEFF Research Database (Denmark)

    Christensen, Laurids Siig; Brehm, Katharina E.; Skov, Julia

    2011-01-01

    Exhaled air of individual cattle infected experimentally with foot-and-mouth disease virus (FMDV) was sampled to assess the feasibility of a rapid, non-invasive general screening approach for identifying sources of FMDV infection. The air sampler used was a handheld prototype device employing...... of FMDV in cattle infected experimentally. Detection in exhaled air from individual cattle was compared to FMDV detection in serum and saliva for 3 different strains of FMDV (O1/Manisa/69, C/Oberbayern/FRG/1960 and SAT1/Zimbawe/1989). Detection of FMDV in exhaled air was possible for all strains of FMDV...

  1. Molecularly imprinted solid phase microextraction fiber for trace analysis of catecholamines in urine and serum samples by capillary electrophoresis.

    Science.gov (United States)

    Zhang, Xinfeng; Xu, Shuxia; Lim, Jae-Min; Lee, Yong-Ill

    2012-09-15

    A selective and flexible monolithic moleculary imprinted polymer (MIP) fiber was developed in batch for solid phase microextraction (SPME) of catecholamines (CAs), i.e., dopamine (DA), epinephrine (E) and norepinephrine (NE), and coupled with capillary electrophoresis (CE) for trace analysis of urine and serum samples. The polymer fiber was synthesized in-situ simply using a flexible capillary as a mold and the polymerization protocols and SPME experimental conditions were examined in detail. The reproducibility of fiber to fiber fabrication (n=5) was in range of 5.9-9.8% for three CAs. The fiber also shows high stability without any deterioration of extraction performance after 30 times use. Under the established optimum conditions, the limits of detection for DA, E, and NE were 7.4, 4.8, and 7.1 nmol L(-1), respectively, with the enhancement factor over 100 after MIP-SPME. The specific selectivity to three CAs was discovered with the developed MIP fibers compared with non-imprinted polymer (NIP) fiber. Finally, the MIP fibers were successfully applied for selective extraction of CAs in urine and serum samples with the relative recoveries ranging from 85% to 103%. The fabricated MIP-fibers were promising in preparation of biological samples in batch followed by CE-UV detection. Copyright © 2012 Elsevier B.V. All rights reserved.

  2. The quality of analytical determinations of copper and zinc in serum. A collaborative study in a sample of Italian laboratories.

    Science.gov (United States)

    Patriarca, M; Menditto, A; Muran, A; Morisi, G

    1993-12-01

    An external quality assurance scheme for Cu and Zn analysis in serum was carried out in Italy between 1989 and 1990. It included: preparation of control materials by the organizing centre; distribution of control samples to the participants according to a random selection; elaboration of results and evaluation of laboratory performance. Inter-laboratory variability was 16.9% and 16.4%, at concentrations of 0.94 and 1.08 mg/l, for Cu and Zn, respectively. Laboratory performance in terms of percentage of results within the limits of acceptability, improved for Cu, but not for Zn analysis.

  3. Synthesis and Evaluation of a Molecularly Imprinted Polymer for Selective Solid-Phase Extraction of Irinotecan from Human Serum Samples

    Directory of Open Access Journals (Sweden)

    Isabelle Lefebvre-Tournier

    2012-02-01

    Full Text Available A molecularly imprinted polymer (MIP was synthesized by non-covalent imprinting polymerization using irinotecan as template. Methacrylic acid and 4-vinylpyridine were selected as functional monomers. An optimized procedure coupled to LC-PDA analysis was developed for the selective solid-phase extraction of irinotecan from various organic media. A specific capacity of 0.65 µmol•g−1 for the MIP was determined. The high specificity of this MIP was demonstrated by studying the retention behaviour of two related compounds, camptothecin and SN-38. This support was applied for the extraction of irinotecan from human serum samples.

  4. Development and Validation of HPLC Method for Determination of Crocetin, a constituent of Saffron, in Human Serum Samples

    Directory of Open Access Journals (Sweden)

    Amir Hooshang Mohammadpour

    2013-01-01

    Full Text Available Objective(s:The present study reports the development and validation of a sensitive and rapid extraction method beside high performance liquid chromatographic method for the determination of crocetin in human serum. Materials and Methods:The HPLC method was carried out by using a C18 reversed-phase column and a mobile phase composed of methanol/water/acetic acid (85:14.5:0.5 v/v/v at the flow rate of 0.8 ml/min. The UV detector was set at 423 nm and 13-cis retinoic acid was used as the internal standard. Serum samples were pretreated with solid-phase extraction using Bond Elut C18 (200mg cartridges or with direct precipitation using acetonitrile. Results:The calibration curves were linear over the range of 0.05-1.25 µg/ml for direct precipitation method and 0.5-5 µg/ml for solid-phase extraction. The mean recoveries of crocetin over a concentration range of 0.05-5 µg/ml serum for direct precipitation method and 0.5-5 µg/ml for solid-phase extraction were above 70 % and 60 %, respectively. The intraday coefficients of variation were 0.37- 2.6% for direct precipitation method and 0.64 - 5.43% for solid-phase extraction. The inter day coefficients of variation were 1.69 – 6.03% for direct precipitation method and 5.13-12.74% for solid-phase extraction, respectively. The lower limit of quantification for crocetin was 0.05 µg/ml for direct precipitation method and 0.5 µg/ml for solid-phase extraction. Conclusion: The validated direct precipitation method for HPLC satisfied all of the criteria that were necessary for a bioanalytical method and could reliably quantitate crocetin in human serum for future clinical pharmacokinetic study

  5. Anti-Neospora caninum antibodies among dairy cattle in a rural settlement, Paraná, Brazil

    Directory of Open Access Journals (Sweden)

    Shiguedy Katto

    2017-03-01

    Full Text Available The aim of the present study was to evaluate the seroprevalence of antibodies against Neospora caninum, obtained two years apart, among dairy cattle in a rural settlement in southern Brazil. Blood samples from 734 dairy cattle on 41 farms were collected at two different times: in 2012, 406 animals on 30 farms were used; and in 2014, 329 animals on 31 farms. Serum samples were obtained and were used to detect antibodies against N. caninum, by means of the indirect fluorescence assay (IFA. Animals with titers ? 100 were considered positive. The total serum prevalence of anti-N. caninum antibodies was 19.7% (145/736 among all the dairy cattle, comprising 23.1% (94/406 in 2012 and 15.5% (51/329 in 2014. Serum from 91 animals was tested in both trials: 11(12.1% showed positivity in 2012 and 10 (11% in 2014. The variables of age, sex and breed did not show any associations with seropositivity. Thus, we showed that the cattle in this settlement presented high levels of antibodies against N. caninum, and that IFA showed good efficacy for epidemiological studies.

  6. A new treatment by dispersive liquid-liquid microextraction for the determination of parabens in human serum samples.

    Science.gov (United States)

    Vela-Soria, F; Ballesteros, O; Rodríguez, I; Zafra-Gómez, A; Ballesteros, L; Cela, R; Navalón, A

    2013-09-01

    Alkyl esters of p-hydroxybenzoic acid (parabens) are a family of compounds that have been in use since the 1920s as preservatives in cosmetic formulations, with one of the lowest rates of skin problems reported in dermatological patients. However, in the last few years, many scientific publications have demonstrated that parabens are weak endocrine disruptors, meaning that they can interfere with the function of endogenous hormones, increasing the risk of breast cancer. In the present work, a new sample treatment method is introduced based on dispersive liquid-liquid microextraction for the extraction of the most commonly used parabens (methyl-, ethyl-, propyl-, and butylparaben) from human serum samples followed by separation and quantification using ultrahigh performance liquid chromatography-tandem mass spectrometry. The method involves an enzymatic treatment to quantify the total content of parabens. The extraction parameters (solvent and disperser solvent, extractant and dispersant volume, pH of the sample, salt addition, and extraction time) were accurately optimized using multivariate optimization strategies. Ethylparaben ring (13)C6-labeled was used as surrogate. Limits of quantification ranging from 0.2 to 0.7 ng mL(-1) and an interday variability (evaluated as relative standard deviations) from 3.8 to 11.9 % were obtained. The method was validated using matrix-matched calibration standard and a spike recovery assay. Recovery rates for spiked samples ranged from 96 to 106 %, and a good linearity up to concentrations of 100 ng mL(-1) was obtained. The method was satisfactorily applied for the determination of target compounds in human serum samples.

  7. Determination of tapentadol and tapentadol-O-glucuronide in human serum samples by UPLC-MS/MS.

    Science.gov (United States)

    Hillewaert, Vera; Pusecker, Klaus; Sips, Luc; Verhaeghe, Tom; de Vries, Ronald; Langhans, Manfred; Terlinden, Rolf; Timmerman, Philip

    2015-02-15

    Tapentadol is a novel, centrally acting analgesic with 2 mechanisms of action, MOR agonism and noradrenaline (NA) reuptake inhibition in a single molecule. It is the first member of a new therapeutic class, MOR-NRI. A high throughput liquid chromatography-tandem mass spectrometric (LC-MS/MS) assay was developed and validated for the quantitative analysis of tapentadol and its O-glucuronide metabolite in human serum. Simultaneous quantification was deemed to be challenging because of the large difference in concentrations between tapentadol and its O-glucuronide metabolite in clinical samples. Therefore, a method was established using a common processed sample, but with different injection volumes and chromatographic conditions for each analyte. Tapentadol and tapentadol-O-glucuronide were determined by protein precipitation of 0.100ml of the samples with acetonitrile. The internal standards used are D₆-tapentadol and D₆-tapentadol-O-glucuronide. The validated concentration range was 0.200-200 ng/ml (tapentadol) and 10.0-10,000 ng/ml (tapentadol-O-glucuronide). Chromatographic separation was achieved by gradient elution on a Waters Acquity UPLC BEH C18 (1.7 μm, 2.1 × 50 mm) column, with mobile phase consisting of 0.01 M ammonium formate (adjusted to pH 4 using formic acid) (A) and methanol (B). A separate injection was done for measurement of each analyte, with a different gradient and run time. The analytes were detected by using an electrospray ion source on a triple quadrupole mass spectrometer operating in positive ionization mode. The run time was 1.6 min for tapentadol and 1.5 min for tapentadol-O-glucuronide. The high sensitivity and acceptable performance of the assay allowed its application to the analysis of serum samples in clinical trials. The validated method was used for analysis of tapentadol in over 17,000 samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Regulation of hepcidin: insights from biochemical analyses on human serum samples.

    Science.gov (United States)

    Kemna, Erwin H J M; Kartikasari, April E R; van Tits, Lambertus J H; Pickkers, Peter; Tjalsma, Harold; Swinkels, Dorine W

    2008-01-01

    Knowledge of hepcidin regulation is foremost gained by in vitro studies. We aimed to translate this knowledge into the human in vivo situation. Therefore, we measured serum markers as transferrin saturation (TS), soluble transferrin receptor (sTfR), and C-reactive protein (CRP) in parallel with hepcidin and prohepcidin in patients with iron metabolism disorders and controls. To assess sTfR as erythropoietic activity-associated factor in hepcidin regulation, we studied its influence on hepcidin expression in HepG2 cells. Results showed that sTfR highly associates with erythropoietic activity that strongly interfered with the iron store regulation of hepcidin. HepG2 expression results display an inverse association between hepcidin and sTfR. Inflammation was strongly related to increased hepcidin levels regardless of the iron store and erythropoietic activity status. In contrast, prohepcidin failed to correlate to any other parameter. In conclusion, these studies verify that previous conclusions based on in vitro studies on hepcidin regulation are also likely to apply to human patients. This is underscored by a simple algorithm, based on parameters reflecting the main regulating pathways, that accurately predict the actual measured hepcidin levels. Future studies are needed to validate the combined utility of this predictive algorithm together with actual measured hepcidin levels in clinical diagnosis.

  9. Prevalence and distribution of Neospora caninum in water buffalo (Bubalus bubalis) and cattle in the Northern Territory of Australia.

    Science.gov (United States)

    Neverauskas, Claudia E; Nasir, Amar; Reichel, Michael P

    2015-10-01

    The seroprevalence of Neospora caninum infection in water buffalo (Bubalus bubalis) and domestic cattle in the Northern Territory (NT) of Australia has never been determined. A total of 480 serum samples from water buffalo and 192 serum samples from cattle, collected by the NT Government from 1993 through to 2001, at 18 different survey sites throughout the Northern Territory were tested by commercial ELISA for anti-N. caninum antibodies. The water buffalo samples demonstrated a seroprevalence of 88.3% (95% CI ± 2.9%), while 31.8% (±6.1%) of the cattle sera tested positive for N. caninum antibodies. Individual buffalo from the same herd, sampled over years, showed considerable fluctuations in S/P ratios. Overall, seropositivity was consistent across buffalo herds, and showed a slight decline over the years. The study presents evidence for the first time that N. caninum infection in water buffalo in the Northern Territory is a highly endemic and that infection rates are higher than those for cattle. This is important for an understanding of any potential sylvatic life cycle of N. caninum in Northern Australia. This survey also tests cattle from that territory for the first time for evidence of N. caninum infection and makes an important contribution to the understanding of disease management issues for the beef industry in the region. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  10. Anatomopatologia de amostras de bovinos alimentados com glicerol Anatomic-pathological samples of cattle fed with glycerol

    Directory of Open Access Journals (Sweden)

    Jean Pacheco Leão

    2012-07-01

    possible toxic action of crude glycerin and methanol by pathological examinations of the central nervous system (CNS, liver, rumen, small intestine and kidney from 24 feedlot cattle for 84 days, feed containing up to 24% of crude glycerin in dry matter (DM presenting in its composition 80.35% glycerol, and <0.01% methanol. There were no pathological changes observed in dairy cows and steers up to 24% of glycerol inclusion based on the diet DM. The total weight gain and feed conversion were not affected by inclusion of glycerol. We conclude that crude glycerin can be included up to 24% of total DM in the diets of steers and dairy cows without compromising animal performance or display toxicity.

  11. Identification of Reliable Reference Genes for Quantification of MicroRNAs in Serum Samples of Sulfur Mustard-Exposed Veterans

    Science.gov (United States)

    Gharbi, Sedigheh; Shamsara, Mehdi; Khateri, Shahriar; Soroush, Mohammad Reza; Ghorbanmehr, Nassim; Tavallaei, Mahmood; Nourani, Mohammad Reza; Mowla, Seyed Javad

    2015-01-01

    Objective In spite of accumulating information about pathological aspects of sulfur mustard (SM), the precise mechanism responsible for its effects is not well understood. Circulating microRNAs (miRNAs) are promising biomarkers for disease diagnosis and prognosis. Accurate normalization using appropriate reference genes, is a critical step in miRNA expression studies. In this study, we aimed to identify appropriate reference gene for microRNA quantification in serum samples of SM victims. Materials and Methods In this case and control experimental study, using quantitative real-time polymerase chain reaction (qRT-PCR), we evaluated the suitability of a panel of small RNAs including SNORD38B, SNORD49A, U6, 5S rRNA, miR-423-3p, miR-191, miR-16 and miR-103 in sera of 28 SM-exposed veterans of Iran-Iraq war (1980-1988) and 15 matched control volunteers. Different statistical algorithms including geNorm, Normfinder, best-keeper and comparative delta-quantification cycle (Cq) method were employed to find the least variable reference gene. Results miR-423-3p was identified as the most stably expressed reference gene, and miR- 103 and miR-16 ranked after that. Conclusion We demonstrate that non-miRNA reference genes have the least stabil- ity in serum samples and that some house-keeping miRNAs may be used as more reliable reference genes for miRNAs in serum. In addition, using the geometric mean of two reference genes could increase the reliability of the normalizers. PMID:26464821

  12. Comparação de kits ELISA® comerciais para anticorpos no soro e leite com um teste coproparasitológico em bovinos naturalmente infectados por Fasciola hepatica Comparison of comercial® ELISA kits for antibodies in serum and milk with a fecal test in cattle naturally infected with Fasciola hepatica

    Directory of Open Access Journals (Sweden)

    Cíntia das C. Bernardo

    2013-01-01

    Full Text Available A fasciolose é uma enfermidade causada por um trematoda que acomete o fígado principalmente de ruminantes domésticos, podendo parasitar o homem e seu diagnóstico é realizado rotineiramente por exames coproparasitológicos. O objetivo do presente estudo foi comparar kits comerciais de ELISA para anticorpos no soro e leite com um teste coproprarasitológico em bovinos naturalmente infectados por Fasciola hepatica. Foram coletadas amostras de fezes (92 sangue (92 e leite (43 de bovinos provenientes de propriedades de gado leiteiro do município de Jerônimo Monteiro, sul do Estado do Espírito Santo. As amostras de fezes coletadas foram processadas pela técnica de sedimentação fecal para ovos de F. hepatica, utilizada como padrão ouro para as análises. Amostras de sangue e de leite foram processadas segundo a orientação do fabricante dos respectivos Kits ELISA comerciais testados. Utilizou-se o c² de McNemar para comparação estatística e calcularam-se a sensibilidade e especificidade, valores preditivos e kappa. Os resultados obtidos mostraram que as frequências de positividade pelo uso dos kits ELISA comerciais de soro e de leite diferiram significativamente (pThe fascioliasis is a disease caused by a trematode that affects the liver mainly of domestic ruminants and can also parasite man; its diagnosis is routinely done by coprological methods. The aim of this study was to compare commercial ELISA kits for antibodies in serum and milk with a coprological test in cattle naturally infected by Fasciola hepatica. We collected fecal, blood and milk samples from cattle in the municipality of Jerônimo Monteiro, southern Espírito Santo state. The fecal samples were processed by the fecal egg sedimentation for F. hepatica, which is used as a gold standard for analyzis. Blood (92 and milk (43 samples were processed according to the manufacturer instructions of the respective commercial ELISA kits tested. We used the McNemar chi-square for

  13. Particle size distribution and chemical composition of total mixed rations for dairy cattle: water addition and feed sampling effects.

    Science.gov (United States)

    Arzola-Alvarez, C; Bocanegra-Viezca, J A; Murphy, M R; Salinas-Chavira, J; Corral-Luna, A; Romanos, A; Ruíz-Barrera, O; Rodríguez-Muela, C

    2010-09-01

    Four dairy farms were used to determine the effects of water addition to diets and sample collection location on the particle size distribution and chemical composition of total mixed rations (TMR). Samples were collected weekly from the mixing wagon and from 3 locations in the feed bunk (top, middle, and bottom) for 5 mo (April, May, July, August, and October). Samples were partially dried to determine the effect of moisture on particle size distribution. Particle size distribution was measured using the Penn State Particle Size Separator. Crude protein, neutral detergent fiber, and acid detergent fiber contents were also analyzed. Particle fractions 19 to 8, 8 to 1.18, and 19 mm was greater than recommended for TMR, according to the guidelines of Cooperative Extension of Pennsylvania State University. The particle size distribution in April differed from that in October, but intermediate months (May, July, and August) had similar particle size distributions. Samples from the bottom of the feed bunk had the highest percentage of particles retained on the 19-mm sieve. Samples from the top and middle of the feed bunk were similar to that from the mixing wagon. Higher percentages of particles were retained on >19, 19 to 8, and 8 to 1.18 mm sieves for wet than dried samples. The reverse was found for particles passing the 1.18-mm sieve. Mean particle size was higher for wet than dried samples. The crude protein, neutral detergent fiber, and acid detergent fiber contents of TMR varied with month of sampling (18-21, 40-57, and 21-34%, respectively) but were within recommended ranges for high-yielding dairy cows. Analyses of TMR particle size distributions are useful for proper feed bunk management and formulation of diets that maintain rumen function and maximize milk production and quality. Water addition may help reduce dust associated with feeding TMR. Copyright (c) 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  14. Detection of the Inflammation Biomarker C-Reactive Protein in Serum Samples: Towards an Optimal Biosensor Formula

    Directory of Open Access Journals (Sweden)

    Wellington M. Fakanya

    2014-10-01

    Full Text Available The development of an electrochemical immunosensor for the biomarker, C-reactive protein (CRP, is reported in this work. CRP has been used to assess inflammation and is also used in a multi-biomarker system as a predictive biomarker for cardiovascular disease risk. A gold-based working electrode sensor was developed, and the types of electrode printing inks and ink curing techniques were then optimized. The electrodes with the best performance parameters were then employed for the construction of an immunosensor for CRP by immobilizing anti-human CRP antibody on the working electrode surface. A sandwich enzyme-linked immunosorbent assay (ELISA was then constructed after sample addition by using anti-human CRP antibody labelled with horseradish peroxidase (HRP. The signal was generated by the addition of a mediator/substrate system comprised of 3,3,5',5'-Tetramethylbenzidine dihydrochloride (TMB and hydrogen peroxide (H2O2. Measurements were conducted using chronoamperometry at −200 mV against an integrated Ag/AgCl reference electrode. A CRP limit of detection (LOD of 2.2 ng·mL−1 was achieved in spiked serum samples, and performance agreement was obtained with reference to a commercial ELISA kit. The developed CRP immunosensor was able to detect a diagnostically relevant range of the biomarker in serum without the need for signal amplification using nanoparticles, paving the way for future development on a cardiac panel electrochemical point-of-care diagnostic device.

  15. A generic sample preparation approach for LC–MS/MS bioanalysis of therapeutic monoclonal antibodies in serum applied to Infliximab

    Directory of Open Access Journals (Sweden)

    Anne J. Kleinnijenhuis

    2015-01-01

    Full Text Available In this study, we developed a generic bioanalytical workflow providing sensitive, specific, and accurate absolute quantification of therapeutic monoclonal antibodies in serum. The workflow involves magnetic beads coated with protein A to pull-down therapeutic monoclonal antibodies with affinity for protein A from the biological matrix, followed by tryptic digestion and LC-MS/MS quantification of a unique signature peptide, considering of course the matrix of interest and other present mAbs, if applicable. The feasibility of this approach was demonstrated for Infliximab (trade name Remicade in rat serum. The assigned signature peptide was monitored in the selected reaction monitoring (SRM mode. Assay variability was determined to be below 20%, except at the QC low level, which was provided through optimization of the sample preparation and monitoring of the LC-MS/MS using a stable isotope labeled signature peptide as internal standard. The 100 ng/ml lower limit of quantification using only 25 μl sample volume, is generally considered as sufficient for pharmaceutical development purposes for monoclonal antibodies.

  16. Sample dilution : A methodological pitfall in the measurement of tissue but not serum ACE-activity

    NARCIS (Netherlands)

    Koiter, J; Navis, G; de Jong, PE; van Gilst, WH; de Zeeuw, D

    Many tissue ACE-assays suffer from underestimation of the ACE-activity at low sample dilutions. However, measurement of ACE-activity as the amount of hippuric acid produced by cleavage of the commonly used substrate hippuryl-histidyl-leucine might circumvent this problem. In this study, we

  17. Cattle rabies vaccination--A longitudinal study of rabies antibody titres in an Israeli dairy herd.

    Science.gov (United States)

    Yakobson, Boris; Taylor, Nick; Dveres, Nelli; Rozenblut, Shira; Tov, Boris Even; Markos, Majid; Gallon, Nadav; Homer, David; Maki, Joanne

    2015-09-01

    In contrast to many regions of the world where rabies is endemic in terrestrial wildlife species, wildlife rabies has been controlled in Israel by oral rabies vaccination programs, but canine rabies is re-emerging in the northern area of the Golan Heights. From 2009 to 2014 there were 208 animal rabies cases in Israel; 96 (46%) were considered introduced primary cases in dogs, triggering 112 secondary cases. One third (37/112) of the secondary cases were in cattle. Rabies vaccination is voluntary for cattle in Israel, except those on public exhibit. Rabies vaccination schedules for cattle vary based on farm practices and perception of risk. In this study 59 cattle from a dairy farm which routinely vaccinates against rabies were assigned into six groups according to age and vaccination histories. Four groups contained adult cows which had received one previous rabies vaccination, one group of adults had received two previous vaccinations, and one group was unvaccinated calves. Serum samples were collected and the cows were vaccinated with a commercial rabies vaccine. Sera were again collected 39 days later and the calf group re-vaccinated and re-sampled 18 days later. Sera were analyzed for the presence of rabies virus neutralizing antibodies using the rapid immunofluorescent antibody test. Cattle with antibody titres ≥ 0.5 IU/ml were considered to be protected against rabies. Twenty-six of 27 adult cattle (96%) vaccinated once at less than five months old did not have protective titres. Sixty percent (6/10) cattle vaccinated once at around six months of age did have adequate titres. Cattle previously vaccinated twice (n=10; 100%) with an 18 month interval between inoculations, had protective titres and protective antibody titres following booster vaccination (n=51; 100%). The anamnestic response of cattle to a killed rabies vaccine was not affected by the time interval between vaccinations, which ranged from 12 to 36 months. These results suggest that calves from

  18. Comprehensive metabolic characterization of serum osteocalcin action in a large non-diabetic sample

    DEFF Research Database (Denmark)

    Entenmann, Lukas; Pietzner, Maik; Artati, Anna

    2017-01-01

    Recent research suggested a metabolic implication of osteocalcin (OCN) in e.g. insulin sensitivity or steroid production. We used an untargeted metabolomics approach by analyzing plasma and urine samples of 931 participants using mass spectrometry to reveal further metabolic actions of OCN. Sever...... study provides a read-out of metabolic actions of OCN. However, most of the associations were weak arguing for a limited role of OCN in whole-body metabolism....

  19. Effect of fetal and adult bovine serum on pyocyanin production in Pseudomonas aeruginosa isolated from clinical and soil samples.

    Science.gov (United States)

    Moayedi, Aylin; Nowroozi, Jamileh; Sepahy, Abbas Akhavan

    2017-12-01

    Pyocyanin is a blue-greenish redox-active pigment, produced by Pseudomonas aeruginosa, with a wide range of biological and biotechnological applications. Pyocyanin biosynthesis is regulated by the quorum-sensing (QS) system in which the expression of QS genes and QS-controlled virulence genes may be affected by serum as a complex medium. In the current study, effects of adult bovine serum (ABS) and fetal bovine serum (FBS) on the production of pyocyanin were examined in order to develop it. The presence of pyocyanin-producing specific genes and proteins in clinical and soil isolates of P. aeruginosa was confirmed using PCR and SDS-PAGE. Isolates were inoculated to media containing different concentrations of complement-active/-inactivated ABS or FBS and pyocyanin concentration was measured by spectrophotometry. Extracted pigment was characterized by using UV-Visible spectrophotometry. Titration of ABS antibodies against studied isolates was performed by the tube agglutination test. Adding ABS to P. aeruginosa culture medium decreased pyocyanin production compared to the control, while its production increased in FBS-containing media (113.21±2.581 vs. 55.26±0.827 μg.ml-1 and 126.80±2.036 vs. 30.56±0.382 μg.ml-1 of C11 and E8 pyocyanin concentration in the presence of 10% FBS vs. control, respectively). In this study, due to the presence of inhibitors such as complement proteins and antibodies in ABS samples, the use of FBS devoid of antibodies was effective to increase pyocyanin production in studied isolates.

  20. Comparison of diagnostic methods to detect Histoplasma capsulatum in serum and blood samples from AIDS patients

    Science.gov (United States)

    da Silva, Marcos Vinicius; Criado, Paulo Ricardo; Luiz, Olinda do Carmo; Vicentini, Adriana Pardini

    2018-01-01

    Background Although early and rapid detection of histoplasmosis is essential to prevent morbidity and mortality, few diagnostic tools are available in resource-limited areas, especially where it is endemic and HIV/AIDS is also epidemic. Thus, we compared conventional and molecular methods to detect Histoplasma capsulatum in sera and blood from HIV/AIDS patients. Methodology We collected a total of 40 samples from control volunteers and patients suspected of histoplasmosis, some of whom were also infected with other pathogens. Samples were then analyzed by mycological, serological, and molecular methods, and stratified as histoplasmostic with (group I) or without AIDS (group II), uninfected (group III), and infected with HIV and other pathogens only (group IV). All patients were receiving treatment for histoplasmosis and other infections at the time of sample collection. Results Comparison of conventional methods with nested PCR using primers against H. capsulatum 18S rRNA (HC18S), 5.8S rRNA ITS (HC5.8S-ITS), and a 100 kDa protein (HC100) revealed that sensitivity against sera was highest for PCR with HC5.8S-ITS, followed by immunoblotting, double immunodiffusion, PCR with HC18S, and PCR with HC100. Specificity was equally high for double immunodiffusion, immunoblotting and PCR with HC100, followed for PCR with HC18S and HC5.8-ITS. Against blood, sensitivity was highest for PCR with HC5.8S-ITS, followed by PCR with HC18S, Giemsa staining, and PCR with HC100. Specificity was highest for Giemsa staining and PCR with HC100, followed by PCR with HC18S and HC5.8S-ITS. PCR was less efficient in patients with immunodeficiency due to HIV/AIDS and/or related diseases. Conclusion Molecular techniques may detect histoplasmosis even in cases with negative serology and mycology, potentially enabling early diagnosis. PMID:29342162

  1. Retrospective analysis of synthetic cannabinoids in serum samples--epidemiology and consumption patterns.

    Science.gov (United States)

    Jaenicke, Nathalie J; Pogoda, Werner; Paulke, Alexander; Wunder, Cora; Toennes, Stefan W

    2014-09-01

    Herbal mixtures contain synthetic cannabinoids, which can cause severe intoxications. Due to the great variety and the changing spectrum of substances on the drug market, prevalence data are limited, and data on prevalence rates of synthetic cannabinoids in forensic cases are not available. The present study was performed to survey the prevalence of synthetic cannabinoids in cases of traffic and criminal offences in the German state Hesse in 2010. The applied analytical method covered all synthetic cannabinoids on the drug market at that time, and with 20% of the blood samples (422 out of 2201) a representative number was reanalyzed. In twelve samples synthetic cannabinoids were identified and a prevalence of 2.8% was estimated. Consumption patterns showed predominantly cases of multi-drug consumption (10 cases); the combination with cannabis or alcohol was frequent (four cases each). The observed deficits were moderate with the exception of aggravation of paranoia in one case. The symptoms were either compatible with the effects of cannabinoid agonists or attributable to alcohol or other drugs found in the blood samples. Our current analytical strategy is to perform such analyses only in cases where use is suspected or where symptoms are not explained by routine toxicological analyses. Hence, the positive rate is rather low highlighting the need to keep up with the developments on the drug market and to establish sensitive screening methods covering a broad range of substances that can be updated fast, e.g., relying on collections of mass spectrometric reference data. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  2. Chemiluminescence lateral flow immunoassay cartridge with integrated amorphous silicon photosensors array for human serum albumin detection in urine samples.

    Science.gov (United States)

    Zangheri, Martina; Di Nardo, Fabio; Mirasoli, Mara; Anfossi, Laura; Nascetti, Augusto; Caputo, Domenico; De Cesare, Giampiero; Guardigli, Massimo; Baggiani, Claudio; Roda, Aldo

    2016-12-01

    A novel and disposable cartridge for chemiluminescent (CL)-lateral flow immunoassay (LFIA) with integrated amorphous silicon (a-Si:H) photosensors array was developed and applied to quantitatively detect human serum albumin (HSA) in urine samples. The presented analytical method is based on an indirect competitive immunoassay using horseradish peroxidase (HRP) as a tracer, which is detected by adding the luminol/enhancer/hydrogen peroxide CL cocktail. The system comprises an array of a-Si:H photosensors deposited on a glass substrate, on which a PDMS cartridge that houses the LFIA strip and the reagents necessary for the CL immunoassay was optically coupled to obtain an integrated analytical device controlled by a portable read-out electronics. The method is simple and fast with a detection limit of 2.5 mg L -1 for HSA in urine and a dynamic range up to 850 mg L -1 , which is suitable for measuring physiological levels of HSA in urine samples and their variation in different diseases (micro- and macroalbuminuria). The use of CL detection allowed accurate and objective analyte quantification in a dynamic range that extends from femtomoles to picomoles. The analytical performances of this integrated device were found to be comparable with those obtained using a charge-coupled device (CCD) as a reference off-chip detector. These results demonstrate that integrating the a-Si:H photosensors array with CL-LFIA technique provides compact, sensitive and low-cost systems for CL-based bioassays with a wide range of applications for in-field and point-of-care bioanalyses. Graphical Abstract A novel integrated portable device was developed for direct quantitative detection of human serum albumin (HSA) in urine samples, exploiting a chemiluminescence lateral flow immunoassay (LFIA). The device comprises a cartridge that holds the LFIA strip and all the reagents necessary for the analysis, an array of amorphous silicon photosensors, and a custom read-out electronics.

  3. [Differences in Measured Values among Homogenous Assay Reagents of LDL-C in LP-X Positive Serum Samples].

    Science.gov (United States)

    Abe, Misako; Kurosawa, Hideo; Sato, Ryo; Ito, Kumie; Tomono, Yoshiharu; Manita, Daisuke; Hirowatari, Yuji; Yoshida, Hiroshi

    2015-03-01

    The LDL-C level measures with homogeneous (direct) assays in almost of clinical laboratories. Several reports however showed differences in measured values among the assay reagents. We investigated the differences in LDL-C values among direct assays and Friedewald formula (F-f) in 58 LP-X positive serum samples from jaundice patients by comparing LDL-C values measured by anion-exchange chromatography (AEX-HPLC), largely comparable to ultracentrifugation method. Changes in LDL-C values during the treatment of 8 patients were also investigated. Direct assay reagents from Sekisui Medical (S-r), Denka-Seiken (D-r), Wako Chemical (W-r), and Kyowa Medics (K-r) were used for comparison. F-f, S-r, and D-r correlated with AEX-HPLC with r values 0.6. Two samples in which F-f values provided 500 mg/dL plus bias to AEX-HPLC (LDL-C value of 220 mg/dL) demonstrated increased levels of IDL-C before treatment. LDL-C values (S-r and D-r) of the 2 samples were relatively high and near to F-f data while LDL-C values (W-r and K-r) were relatively low and close to AEX-HPLC data. The jaundice treatment decreased LDL-C values (S-r and D-r) and converged to 220 mg/dL, indicating that S-r and D-r might react markedly to IDL. These changes were consistent with decreases in serum free cholesterol and phospholipid in support of LP-X. By contrast, W-r and K-r data showed upward tendency and also converged to 220 mg/dL. These results suggest that LDL-C direct assay reagents would be classified into 2 groups with respect to the reagent reactivity to LP-X.

  4. On-line sample cleanup and enrichment chromatographic technique for the determination of ambroxol in human serum.

    Science.gov (United States)

    Emara, Samy; Kamal, Maha; Abdel Kawi, Mohamed

    2012-02-01

    A sensitive and efficient on-line clean up and pre-concentration method has been developed using column-switching technique and protein-coated µ-Bondapak CN silica pre-column for quantification of ambroxol (AM) in human serum. The method is performed by direct injection of serum sample onto a protein-coated µ-Bondapak CN silica pre-column, where AM is pre-concentrated and retained, while proteins and very polar constituents are washed to waste using a phosphate buffer saline (pH 7.4). The retained analyte on the pre-column is directed onto a C(18) analytical column for separation, with a mobile phase consisting of a mixture of methanol and distilled deionized water (containing 1% triethylamine adjusted to pH 3.5 with ortho-phosphoric acid) in the ratio of 50:50 (v/v). Detection is performed at 254 nm. The calibration curve is linear over the concentration range of 12-120 ng/mL (r(2) = 0.9995). The recovery, selectivity, linearity, precision, and accuracy of the method are convenient for pharmacokinetic studies or routine assays.

  5. Reference values of coplanar and non-coplanar PCBs in serum samples from two Italian population groups.

    Science.gov (United States)

    Turci, Roberta; Finozzi, Enrico; Catenacci, Giovanni; Marinaccio, Alessandro; Balducci, Claudio; Minoia, Claudio

    2006-04-10

    The main goal of this study is to establish the reference values of individual Polychlorinated biphenyl (PCB) congeners in non-occupationally exposed subjects. Since the PCB pattern in human serum is related to the living area, two different population groups from North and Central Italy, were compared. Serum concentrations of both coplanar and non-coplanar PCB congeners were measured by using gas chromatography coupled with low-resolution mass spectrometry (HRGC-LRMS). A fast and reliable method for the determination of 60 congeners had been previously validated. Its reliability was further verified by using high-resolution mass spectrometry. Thirty-one congeners out of 60 were found at detectable concentrations in at least one sample. The mean value for total PCBs was found to be 2.48 and 3.93 microg/L for the two population groups. Eight dioxin-like PCBs were detected. In accordance with the findings from the literature, the most abundant congeners were found to be 153, 138, 180, and 170. Both univariate and multivariate analysis showed that age is a significant determinant of PCB concentrations. The correlation increased with increasing chlorination. Slight differences in the PCB pattern were observed in the two population groups.

  6. Potential bacterial core species associated with digital dermatitis in cattle herds identified by molecular profiling of interdigital skin samples

    DEFF Research Database (Denmark)

    Weiss Nielsen, Martin; Strube, Mikael Lenz; Isbrand, Anastasia

    2016-01-01

    of different molecular methods. Deep sequencing of the 16S rRNA gene variable regions V1–V2 showed that Treponema, Mycoplasma, Fusobacterium and Porphyromonas were the genera best differentiating the DD samples from the controls. Additional deep sequencing analysis of the most abundant genus, Treponema......, targeting another variable region of the 16S rRNA gene, V3–V4, identified 15 different phylotypes, among which Treponema phagedenis-like and Treponema refringens-like species were the most abundant. Although the presence of Treponema spp., Fusobacterium necrophorum and Porphyromonas levii was confirmed...

  7. Development of a Fibrinogen-Specific Sandwich Enzyme-Linked Immunosorbent Assay Microarray Assay for Distinguishing Between Blood Plasma and Serum Samples

    Energy Technology Data Exchange (ETDEWEB)

    Gonzales, Rachel M.; Zhang, Qibin; Zangar, Richard C.; Smith, Richard D.; Metz, Thomas O.

    2011-07-01

    We have developed a fibrinogen-specific sandwich ELISA microarray assay for use in qualitatively distinguishing between blood plasma and serum samples. Three capture antibodies, 49D2, HPA001900, and F8512, were evaluated in conjunction with 1D6 as detection antibody, and the data show that 49D2 and, to a lesser extent, F8512 successfully identify previously unknown plasma and serum samples based upon a ~28-fold difference in signal intensity between the sample types. This assay has utility in rapidly identifying previously archived clinical samples with incomplete annotation in a high throughput manner prior to proteomics analyses.

  8. Common epitopes in LPS of different Enterobacteriaceae are associated with an immune response against Escherichia coli O157 in bovine serum samples.

    Science.gov (United States)

    Navarro, Armando; Eslava, Carlos; García de la Torre, Guadalupe; León, Luis Antonio; Licona, Delia; León, Lemuel; Zarco, Luis Alberto; Cravioto, Alejandro

    2007-11-01

    Epidemiological studies in both humans and animals conducted in Mexico have shown that the isolation frequency of Escherichia coli O157 : H7 is low. In a previous study, IgG antibodies against E. coli O157, O7 and O116 LPS were found in serum samples from children and adults with no previous history of E. coli O157 : H7 infection. The present study was designed to determine whether a similar immune response against E. coli O157 : H7 and other antigenically related bacteria was present in bovine serum samples. A total of 310 serum samples from different herds in Mexico was analysed by microagglutination assays against different enterobacterial antigens, including E. coli O157. Microagglutination assays were positive against E. coli O7 (55 %), O116 (76 %) and O157 (36 %), Escherichia hermannii (15 %), Salmonella enterica serotype Urbana (14 %) and Salmonella enterica subsp. arizonae (40 %). These results were confirmed using a specific ELISA with purified LPS. A positive reaction was observed against the LPS of E. coli O7 (29 %), O116 (12 %) and O157 (22 %), E. hermannii (4 %), Salmonella Urbana (13 %) and S. enterica subsp. arizonae (12 %). Serum absorption studies of positive serum samples indicated the existence of at least three common epitopes shared by the LPS of E. coli O7, O116 and O157, and two others between E. coli O157 and Salmonella Urbana and S. enterica subsp. arizonae. A bactericidal assay against E. coli O157 : H7 using 31 bovine serum samples was performed, and 22 (71 %) of these serum samples gave positive results. The data demonstrated that bovine serum showed a response against different enterobacteria, including E. coli O157, and that this response could be due to the presence of shared epitopes in the LPS of these organisms.

  9. Seroprevalence of some bovine viral respiratory diseases among non vaccinated cattle in Saudi Arabia

    Directory of Open Access Journals (Sweden)

    Mohamed Abd El Fatah Mahmoud

    2013-02-01

    Full Text Available Aim: Four viral pathogens, bovine viral diarrhea virus (BVDV, and bovine herpes virus type 1 (BHV-1, bovine parainfluenza type 3 virus (PI-3V, bovine respiratory syncytial virus (BRSV are mainly associated with bovine respiratory diseases that cause major economic losses in the dairy cattle industry. This study aimed to document exposure of cattle in Saudi Arabia to infectious BVDV, BHV-1, PI-3V and BRSV viruses in non vaccinated cattle in order to obtain epidemiological and immunological information. Materials and Methods: In the present study, 460 random serum samples obtained from non vaccinated cattle in five districts (Riyadh, Eastern Province, Jizan, Najran, Asir of Saudi Arabia between January to March 2011. These samples were tested for presence of antibodies against BVDV, BHV-1, BRSV and PIV-3 by commercial indirect ELISA kits. Results: Our findings displayed that Seropositivity rates were 26 % for BVD, 17.4 % for BHV-1, 69.1 % for PI-3V and 75.6 % for BRSV in the sampled population. In addition, coinfections with more than one virus were considerably common among non-vaccinated dairy cattle. Conclusion: These results indicate that exposure to these agents is common within the study areas. Preventive and control measures against these infectious agents should therefore be adopted. [Vet World 2013; 6(1.000: 1-4

  10. Detection of anti-Neospora caninum antibodies in Iranian native cattle

    Directory of Open Access Journals (Sweden)

    Jamal Gharekhani

    2014-12-01

    Full Text Available Neospora caninum is an Apicomplexan parasite which may cause abortion in cattle. This study investigated occurrences of antibodies against N. caninum in Iranian native cattle. From September 2010 to September 2011, blood samples (n=768 of native cows were collected randomly from different rural regions of Hamedan (n=400 and Kurdistan provinces (n=368 located to the western part of Iran. All the samples were evaluated for IgG antibodies against N. caninum using Enzyme Linked Immunosorbent Assay. The IgG antibodies to N. caninum were found in 14.2% (n=109/768 of serum samples (95% CI: 11.74 - 16.66. There was a significant difference between seropositivity and abortion history (p<0.0001, OR=2.9, unlike to age groups (p=0.105. This is the first report of N. caninum infection in Iranian native cattle. In conclusion, N. caninum is an important factor in abortion in Iranian native cattle. Further comprehensive studies and designing control strategies for improving management in cattle farms are highly recommended.

  11. Mycotoxin Cocktail in the Samples of Oilseed Cake from Early Maturing Cotton Varieties Associated with Cattle Feeding Problems

    Science.gov (United States)

    Yunus, Agha W.; Sulyok, Michael; Böhm, Josef

    2015-01-01

    Cottonseed cake in South East Asia has been associated with health issues in ruminants in the recent years. The present study was carried out to investigate the health issues associated with cottonseed cake feeding in dairy animals in Pakistan. All the cake samples were confirmed to be from early maturing cotton varieties (maturing prior to or during Monsoon). A survey of the resource persons indicated that the feeding problems with cottonseed cake appeared after 4–5 months of post-production storage. All the cake samples had heavy bacterial counts, and contaminated with over a dozen different fungal genera. Screening for toxins revealed co-contamination with toxic levels of nearly a dozen mycotoxins including aflatoxin B1 + B2 (556 to 5574 ppb), ochratoxin A + B (47 to 2335 ppb), cyclopiazonic acid (1090 to 6706 ppb), equisetin (2226 to 12672 ppb), rubrofusarin (81 to 1125), tenuazonic acid (549 to 9882 ppb), 3-nitropropionic acid (111 to 1032 ppb), and citrinin (29 to 359 ppb). Two buffalo calves in a diagnostic feed trial also showed signs of complex toxicity. These results indicate that inappropriate processing and storage of the cake, in the typical conditions of the subcontinent, could be the main contributory factors regarding the low quality of cottonseed cake. PMID:26075378

  12. Mycotoxin Cocktail in the Samples of Oilseed Cake from Early Maturing Cotton Varieties Associated with Cattle Feeding Problems

    Directory of Open Access Journals (Sweden)

    Agha W. Yunus

    2015-06-01

    Full Text Available Cottonseed cake in South East Asia has been associated with health issues in ruminants in the recent years. The present study was carried out to investigate the health issues associated with cottonseed cake feeding in dairy animals in Pakistan. All the cake samples were confirmed to be from early maturing cotton varieties (maturing prior to or during Monsoon. A survey of the resource persons indicated that the feeding problems with cottonseed cake appeared after 4–5 months of post-production storage. All the cake samples had heavy bacterial counts, and contaminated with over a dozen different fungal genera. Screening for toxins revealed co-contamination with toxic levels of nearly a dozen mycotoxins including aflatoxin B1 + B2 (556 to 5574 ppb, ochratoxin A + B (47 to 2335 ppb, cyclopiazonic acid (1090 to 6706 ppb, equisetin (2226 to 12672 ppb, rubrofusarin (81 to 1125, tenuazonic acid (549 to 9882 ppb, 3-nitropropionic acid (111 to 1032 ppb, and citrinin (29 to 359 ppb. Two buffalo calves in a diagnostic feed trial also showed signs of complex toxicity. These results indicate that inappropriate processing and storage of the cake, in the typical conditions of the subcontinent, could be the main contributory factors regarding the low quality of cottonseed cake.

  13. The acute phase response of haptoglobin and serum amyloid A (SAA) in cattle undergoing experimental infection with bovine respiratory syncytial virus

    DEFF Research Database (Denmark)

    Heegaard, Peter M. H.; Godson, D.L.; Toussaint, M.J.M.

    2000-01-01

    The ability of a pure virus infection to induce an acute phase protein response is of interest as viral infections are normally considered to be less efficient in inducing an acute phase protein response than bacterial infections. This was studied in a bovine model for infection with bovine...... respiratory syncytial virus (BRSV), analysing the induction of the two most dominant bovine acute phase proteins haptoglobin and serum amyloid A (SAA). Strong and reproducible acute phase responses were detected for both proteins, peaking at around 7-8 days after inoculation of BRSV, while no response...... was seen in mock-inoculated control animals. The serum concentrations reached for SAA and haptoglobin during the BRSV-induced acute phase response were generally the same or higher than previously reported for bacterial infections in calves. The magnitude and the duration of the haptoglobin response...

  14. [Sensitivity to various antibiotics of coagulase-negative staphylococci isolated from samples of milk from Dutch dairy cattle].

    Science.gov (United States)

    Sampimon, O C; Vernooij, J C A; Mevius, D J; Sol, J

    2007-03-15

    During recent years the prevalence of coagulase-negative staphylococci in milk samples from Dutch dairy cows has increased. In 1999 16.2% of the bacteria isolated from milk collected from cows with subclinical mastitis were coagulase-negative staphylococci. In 2004 this proportion was 42.2%. The proportion of coagulase-negative staphylococci of the bacteria isolated from milk samples from cows with clinical mastitis was 7.3% in 1999 and 14.1% in 2004. In this study, the susceptibility of 108 coagulase-negative staphylococci to oxacillin, cefquinome, streptomycin, neomycin, penicillin, and the combination of nafcillin, penicillin, and streptomycin was tested. The isolates were cultured from milk collected from cows with mastitis and typed using the Api-Staph system. Eight species were identified. Staphylococcus chromogenes was the predominant species (41.7%), followed by Staphylococcus xylosus (15.7%) and Staphylococcus simulans (10.2%). With the agar dilution method all strains proved to be sensitive to cefquinome and 90% to oxacillin. Three isolates (2.8%) were mecA-positive. Despite the agar dilution results, these three isolates should be considered resistant to all beta-lactam antibiotics (penicillins, penicillins combined with a beta-lactamase inhibitor and all generations of cephalosporins). In the agar diffusion test, all isolates proved to be sensitive to the combination of nafcillin-penicillin-streptomycin, 99% were sensitive to neomycin and 1% intermediate sensitive, and 95% were sensitive to streptomycin, 4% resistant, and 1% intermediate sensitive. The coagulase-negative staphylococci were highly resistant to penicillin (37.4%), although the level of resistance varied between species, from 0% for Staphylococcus simulans to 100% for Staphylococcus saprophyticus. Because coagulase-negative staphylococci are resistant to several antibiotics, sensitivity testing is important for targeted treatment of mastitis.

  15. Challenges Associated with Sample Preparation for the Analysis of PBDEs in Human Serum.

    Science.gov (United States)

    Siddique, Shabana; Kosarac, Ivana; Kubwabo, Cariton; Harris, Shelley

    2016-01-01

    Polybrominated diphenyl ethers (PBDEs) are used as flame retardants in many applications; however, certain PBDE congeners are persistent, bioaccumulative, and toxic to both humans and the environment. PBDEs have been found in human specimens, and a variety of analytical techniques have been used for their determination in biological matrixes. Nevertheless, obtaining a relatively clean analytical blank sample during PBDE analysis is a big challenge because of the ubiquitous nature of these compounds. Thus, the present study was conducted to compare the PBDE background levels associated with the three most commonly used extraction techniques: liquid-liquid extraction (LLE), SPE, and accelerated solvent extraction (ASE). Conventionally used blank matrixes (HPLC grade water, Milli-Q water, and air) were spiked with internal standards and extracted using LLE, SPE, or ASE. The extracts were analyzed by GC/electron ionization-tandem MS. The ASE method achieved the lowest background levels for nearly all the PBDE congeners analyzed, which may be attributed to the stainless steel and closed-vessel nature of the ASE cells.

  16. Standardizing serum 25-hydroxyvitamin D data from four Nordic population samples using the Vitamin D Standardization Program protocols: Shedding new light on vitamin D status in Nordic individuals

    DEFF Research Database (Denmark)

    Cashman, Kevin D; Dowling, Kirsten G; Škrabáková, Zuzana

    2015-01-01

    protocols for standardizing existing serum 25(OH)D data from national surveys around the world. The objective of the present work was to apply the VDSP protocols to existing serum 25(OH)D data from a Danish, a Norwegian, and a Finnish population-based health survey and from a Danish randomized controlled...... trial. A specifically-selected subset (n 100-150) of bio-banked serum samples from each of the studies were reanalyzed for 25(OH)D by LC-MS/MS and a calibration equation developed between old and new 25(OH)D data, and this equation was applied to the entire data-sets from each study. Compared...... standardization. In conclusion, standardization of serum 25(OH)D concentrations is absolutely necessary in order to compare serum 25(OH)D concentrations across different study populations, which is needed to quantify and prevent vitamin D deficiency....

  17. Detection of antibodies to Plasmodium vivax by indirect immunofluorescence: influence of the geographic origin of antigens and serum samples.

    Science.gov (United States)

    Benzerroug, E H; Demedts, P; Wéry, M

    1986-03-01

    The results of a double-blind serological study of 15 sera sampled in a residual focus of vivax malaria transmission in Algeria, and of 7 sera from patients with slide-proven P. vivax infections acquired in India, are analyzed. The reactivity of each of these serum samples was tested by indirect immunofluorescence using 6 different batches of antigen, including 3 batches of P. vivax antigen prepared with isolates from Zaire (Africa), India and the Solomon Islands, respectively. The geometric mean of reciprocal titers (GMRT) calculated on the 7 sera from proven vivax infections fell from 289.8 using the homologous antigen from the same geographic origin (India) to 48.7 using a homologous (vivax) antigen originating from a different continent (Africa). Among the 15 samples from Algeria, the percentage of seropositives decreased from 100% using the homologous P. vivax antigen originating from the same continent (Africa) to 53.3% using a homologous antigen from India. Two aspects are included in the discussion: in seroepidemiological studies, sensitivity could be improved by the use of a homologous antigen from the same geographic origin; in detection of clinical cases of malaria and species identification based on serology, our results stress the need for caution in interpreting serological titers and for taking into account the geographic origin of the isolates used as antigen.

  18. Sample Preparation Strategies for the Effective Quantitation of Hydrophilic Metabolites in Serum by Multi-Targeted HILIC-MS/MS

    Directory of Open Access Journals (Sweden)

    Elisavet Tsakelidou

    2017-03-01

    Full Text Available The effect of endogenous interferences of serum in multi-targeted metabolite profiling HILIC-MS/MS analysis was investigated by studying different sample preparation procedures. A modified QuEChERS dispersive SPE protocol, a HybridSPE protocol, and a combination of liquid extraction with protein precipitation were compared to a simple protein precipitation. Evaluation of extraction efficiency and sample clean-up was performed for all methods. SPE sorbent materials tested were found to retain hydrophilic analytes together with endogenous interferences, thus additional elution steps were needed. Liquid extraction was not shown to minimise matrix effects. In general, it was observed that a balance should be reached in terms of recovery, efficient clean-up, and sample treatment time when a wide range of metabolites are analysed. A quick step for removing phospholipids prior to the determination of hydrophilic endogenous metabolites is required, however, based on the results from the applied methods, further studies are needed to achieve high recoveries for all metabolites.

  19. Sero-epidemiological survey and risk factors associated with bovine brucellosis among slaughtered cattle in Nigeria

    Directory of Open Access Journals (Sweden)

    Victor O. Akinseye

    2016-03-01

    Full Text Available Bovine brucellosis is endemic in Nigeria; however, limited data exist on nationwide studies and risk factors associated with the disease. Using a cross-sectional sero-epidemiological survey, we determined the prevalence of and risk factors for brucellosis in slaughtered cattle in three geographical regions of Nigeria. Serum samples from randomly selected unvaccinated cattle slaughtered over a period of 3 years (between December 2010 and September 2013 from northern, southern and south-western Nigeria were tested for antibodies to Brucella abortus using the Rose Bengal test. Data associated with risk factors of brucellosis were analysed by Stata Version 12. In all, 8105 cattle were screened. An overall seroprevalence of 3.9% (315/8105 was recorded by the Rose Bengal test, with 3.8%, 3.4% and 4.0% from the northern, southern and south-western regions, respectively. Bivariate analysis showed that cattle screened in northern Nigeria were less likely to be seropositive for antibodies to Brucella spp. than those from south-western Nigeria (odds ratio = 0.94; 95% confidence interval: 0.73–1.22. However, logistic regression analysis revealed that breed ( p = 0.04 and sex ( p £ 0.0001 of cattle were statistically significant for seropositivity to Brucella spp. The study found that brucellosis was endemic at a low prevalence among slaughtered cattle in Nigeria, with sex and breed of cattle being significant risk factors. Considering the public health implications of brucellosis, we advocate coordinated surveillance for the disease among diverse cattle populations in Nigeria, as is carried out in most developed countries.Keywords: Bovine brucellosis, RBT, Epidemiology, Public Health, Nigeria

  20. Sample preparation for mass spectrometric analysis of human serum N-glycans using hydrophilic interaction chromatography-based solid phase extraction.

    Science.gov (United States)

    Cao, Liwei; Zhang, Ye; Chen, Linlin; Shen, Aijin; Zhang, Xingwang; Ren, Shifang; Gu, Jianxin; Yu, Long; Liang, Xinmiao

    2014-09-21

    Expression levels of N-linked glycans derived from human serum glycoproteins have been shown to change during the progression of many diseases. Generally, N-glycans released from human serum proteins co-exist with endogenous serum peptides, salts, and other contaminants. Effective removal of these contaminants is essential to obtain the glycan profile of human serum proteins. Here, we developed a sample preparation method for mass spectrometry (MS) analysis of N-linked glycans derived from human serum glycoproteins based on a zwitterionic hydrophilic material named Click TE-Cys. The high hydrophilicity of Click TE-Cys, resulting from its unique surface structure and charge distribution, facilitated removal of co-existing salts and endogenous serum peptides. Furthermore, the present enrichment approach was handled in parallel, thus saving time. Using this method, a total of 47 unique N-glycans released from human serum proteins were identified. The intrabatch and interbatch coefficients of variation for the 47 N-linked glycans were 8.57% ± 0.96% and 9.22% ± 1.03%, respectively. These results demonstrate that the present method is suitable for fast purification of N-linked glycans derived from human serum glycoproteins, and has potential for clinical application.

  1. Genetic diversity of blastocystis isolated from cattle in khorramabad, iran

    National Research Council Canada - National Science Library

    Badparva, Ebrahim; Sadraee, Javid; Kheirandish, Farnaz

    2015-01-01

    .... The aim of this study was to determine the subtype of Blastocystis in infected cattle. This descriptive cross-sectional study was performed on 196 isolates from cattle stool samples collected from slaughterhouse in Khorramabad city, Iran, in 2012...

  2. Association of Escherichia coli J5-Specific Serum Antibody Responses with Clinical Mastitis Outcome for J5 Vaccinate and Control Dairy Cattle

    Science.gov (United States)

    Wilson, David J.; Mallard, Bonnie A.; Burton, Jeanne L.; Schukken, Ynte H.; Grohn, Yrjo T.

    2009-01-01

    Dairy cattle in two commercial Holstein herds were randomly selected to be vaccinated twice with J5, at approximately 60 days and 28 days before the expected calving date, or to be untreated controls. Based on whether milk production changed following clinical mastitis or whether cows were culled or died within 30 days after onset, 51 mastitis cases were classified as severe or mild. J5-specific antibody responses were evaluated by enzyme-linked immunosorbent assay of all 32 severe and 19 mild cases. The amounts of J5-specific immunoglobulin M (IgM), IgG1, and IgG2 antibodies in sera from the 27 J5 vaccinates were compared with those of the 24 controls. At drying off (before J5 vaccination), all cows had similar amounts of J5-specific antibody. Immediately after calving (approximately 28 days after the second vaccination), J5 vaccinates had significantly higher production of J5-specific IgG1 and IgG2 than controls. When cows were tested following clinical mastitis, none of the three antibody classes differed significantly between the controls and the vaccinates. Vaccinates that contracted Escherichia coli mastitis had 75% less milk loss than controls. The cows that contracted clinical mastitis later in lactation, the unvaccinated controls, and those infected with E. coli had more milk loss following mastitis. The hazards of being culled for all reasons and of being culled for mastitis were significantly lower for J5 vaccinates. Vaccination with J5 was associated with protection against milk production loss and culling following clinical mastitis, and it was also significantly associated with changes in J5-specific IgM, IgG1, and IgG2 antibodies in sera of vaccinated cows. PMID:19052158

  3. Association of Escherichia coli J5-specific serum antibody responses with clinical mastitis outcome for J5 vaccinate and control dairy cattle.

    Science.gov (United States)

    Wilson, David J; Mallard, Bonnie A; Burton, Jeanne L; Schukken, Ynte H; Grohn, Yrjo T

    2009-02-01

    Dairy cattle in two commercial Holstein herds were randomly selected to be vaccinated twice with J5, at approximately 60 days and 28 days before the expected calving date, or to be untreated controls. Based on whether milk production changed following clinical mastitis or whether cows were culled or died within 30 days after onset, 51 mastitis cases were classified as severe or mild. J5-specific antibody responses were evaluated by enzyme-linked immunosorbent assay of all 32 severe and 19 mild cases. The amounts of J5-specific immunoglobulin M (IgM), IgG1, and IgG2 antibodies in sera from the 27 J5 vaccinates were compared with those of the 24 controls. At drying off (before J5 vaccination), all cows had similar amounts of J5-specific antibody. Immediately after calving (approximately 28 days after the second vaccination), J5 vaccinates had significantly higher production of J5-specific IgG1 and IgG2 than controls. When cows were tested following clinical mastitis, none of the three antibody classes differed significantly between the controls and the vaccinates. Vaccinates that contracted Escherichia coli mastitis had 75% less milk loss than controls. The cows that contracted clinical mastitis later in lactation, the unvaccinated controls, and those infected with E. coli had more milk loss following mastitis. The hazards of being culled for all reasons and of being culled for mastitis were significantly lower for J5 vaccinates. Vaccination with J5 was associated with protection against milk production loss and culling following clinical mastitis, and it was also significantly associated with changes in J5-specific IgM, IgG1, and IgG2 antibodies in sera of vaccinated cows.

  4. Evaluation of enzyme-linked immunosorbent assays performed on milk and serum samples for detection of neosporosis and leukosis in lactating dairy cows.

    Science.gov (United States)

    Walsh, Robert B; Kelton, David F; Hietala, Sharon K; Duffield, Todd F

    2013-04-01

    Serum and milk samples from 1229 cows on 22 Ontario dairy farms were individually tested for antibodies specific for bovine leukosis virus (BLV) and Neospora caninum by enzyme-linked immunosorbent assay (ELISA). Antibodies against BLV were present in 361 serum samples (29.4%) and 369 milk samples (30.0%). Comparing the 2 tests, agreement was almost perfect (k = 0.86; 95% CI = 0.83 to 0.90) and the proportions of samples positive were not significantly different (P = 0.56). Both tests identified the same 3 herds free of bovine leukosis virus. Antibodies against N. caninum were detected in 138 serum samples (11.2%), and 111 milk samples (9.0%). Agreement between the 2 tests was moderate (k = 0.52; 95% CI = 0.43 to 0.59). Four herds were free of neosporosis by the serum test, while 10 herds were negative by the milk test. The ELISA on milk samples facilitates sample collection to classify herds free of BLV; the milk N. caninum ELISA was less reliable in predicting herd-level infection.

  5. A cow-level association of ruminal pH on body condition score, serum beta-hydroxybutyrate and postpartum disorders in Thai dairy cattle.

    Science.gov (United States)

    Chaidate, Inchaisri; Somchai, Chanpongsang; Jos, Noordhuizen; Henk, Hogeveen

    2014-09-01

    Subacute ruminal acidosis in dairy cows occurs when ruminal pH is below about 5.5. However, the exact threshold level of ruminal pH affecting cow health is still in debate. This investigation was carried out in 505 cows within 31 farms. The postpartum disorders, including dystocia, retained placenta, anestrus, cystic ovary, metritis, clinical mastitis and lameness, were analyzed. Ruminal pH, serum beta-hydroxy butyrate (SBHB), serum urea nitrogen and body condition score (BCS) were measured once during the 3 to 6 weeks postpartum, while BCS was determined once more at 1 week before calving. Ruminal pH was determined by ruminocentesis technique. The ruminal pH was evaluated to study the association with BCS, SBHB and postpartum disorders using linear regression in a generalized linear mixed model with farm as a random effect. The results show that low ruminal pH was associated with dystocia, metritis and lameness. Moreover, a low ruminal pH can be found in cows with a high loss of BCS after calving and also in cows with low SBHB postpartum. These findings confirmed the feasibility of the ruminocentesis technique and the association of low ruminal pH on various postpartum disorders at the individual cow level. However, the consequences of low ruminal pH on dairy cow health still needs more exploration for a better understanding of the physiological mechanisms. © 2014 Japanese Society of Animal Science.

  6. Unmetabolized Folic Acid Is Detected in Nearly All Serum Samples from US Children, Adolescents, and Adults1234

    Science.gov (United States)

    Pfeiffer, Christine M; Sternberg, Maya R; Fazili, Zia; Yetley, Elizabeth A; Lacher, David A; Bailey, Regan L; Johnson, Clifford L

    2015-01-01

    Background: Serum total folate consists mainly of 5-methyltetrahydrofolate (5-methylTHF). Unmetabolized folic acid (UMFA) may occur in persons consuming folic acid–fortified foods or supplements. Objectives: We describe serum 5-methylTHF and UMFA concentrations in the US population ≥1 y of age by demographic variables and fasting time, stratified by folic acid–containing dietary supplement use. We also evaluate factors associated with UMFA concentrations >1 nmol/L. Methods: Serum samples from the cross-sectional NHANES 2007–2008 were measured for 5-methylTHF (n = 2734) and UMFA (n = 2707) by HPLC–tandem mass spectrometry. Results: In supplement users compared with nonusers, we found significantly higher geometric mean concentrations of 5-methylTHF (48.4 and 30.7 nmol/L, respectively) and UMFA (1.54 and 0.794 nmol/L, respectively). UMFA concentrations were detectable (>0.3 nmol/L) in >95% of supplement users and nonusers, regardless of demographic or fasting characteristics; concentrations differed significantly by age and fasting time, but not by sex and race-ethnicity, both in supplement users and nonusers. The prevalence of UMFA concentrations >1 nmol/L was 33.2% overall and 21.0% in fasting (≥8 h) adults (≥20 y of age). Using multiple logistic regression analysis, UMFA concentrations >1 nmol/L were associated with being older, non-Hispanic black, nonfasting (folic acid intake (diet and supplements), and higher red blood cell folate concentrations. In fasting adults, a decrease in the mean daily alcohol consumption was also associated with increased odds of UMFA concentrations >1 nmol/L. Conclusions: UMFA detection was nearly ubiquitous, and concentrations >1 nmol/L were largely but not entirely explained by fasting status and by total folic acid intake from diet and supplements. These new UMFA data in US persons ≥1 y of age provide much-needed information on this vitamer in a fortified population with relatively high use of dietary supplements

  7. Seroprevalence of Neospora Caninum Infection in Dairy Cattle in Tabriz, Northwest Iran

    Directory of Open Access Journals (Sweden)

    Gh Moghaddam

    2011-09-01

    Full Text Available Background: The aim of this study was to determine the seroprevalence of antibody to Neospora can­inum in healthy and aborted dairy cattle in Tabriz, capital of East-Azarbaijan in northwest of Iran.Methods: In this cross-sectional study serum samples were collected from 266 healthy and ab­orted Holestein-Feriesisnc cows from September 2008 to August 2009. The sera were analyzed to de­tect of antibody against N. caninum using the commercially ELISA kit.Results: Seroprevalence of antibody to N. caninum was 10.5% in Tabriz dairy cattle. Also the abortion rate in all cattle sampled was 33.6% but percentage of seropositive aborted cattle was 18.4%.Conclusion: Neosporosis could be one of the possible causes of abortion in dairy cattle in Tabriz and regarding the distribution in dogs as definitive host for the parasite, further studies in dog and cat­tle are recommended.

  8. Brominated flame retardants in the hair and serum samples from an e-waste recycling area in southeastern China: the possibility of using hair for biomonitoring.

    Science.gov (United States)

    Liang, Si; Xu, Feng; Tang, Weibiao; Zhang, Zheng; Zhang, Wei; Liu, Lili; Wang, Junxia; Lin, Kuangfei

    2016-08-01

    Hair samples and paired serum samples were collected from e-waste and urban areas in Wenling of Zhejiang Province, China. The PBDE and DBDPE concentrations in hair and serum samples from e-waste workers were significantly higher than those of non-occupational residents and urban residents. BDE209 was the dominating BFRs in hair and serum samples from the e-waste area, while DBDPE was the major BFRs from the urban area. Statistically significant correlations were observed between hair level and serum level for some substances (BDE209, DBDPE, BDE99, BDE47, BDE28, and BDE17), although the PBDE congener profiles in hair were different from those in the serum. A statistically significant positive correlation between the PBDE concentrations and the working age, as well as gender difference, was observed in e-waste workers. Different sources of PBDEs and DBDPE in three groups were identified by principal component analysis and spearman correlation coefficient. Hair is suggested to be a useful matrix for biomonitoring the PBDE exposure in humans.

  9. Comparison of the solid phase and liquid-liquid extraction methods for methadone determination in human serum and whole blood samples using gas chromatography/mass spectrometry.

    Science.gov (United States)

    Bratinčević, Maja Veršić; Visković, Tanja; Sutlović, Davorka

    2017-12-20

    The aim of this study was to determine the optimal biological sample and the optimal extraction technique for monitoring methadone concentrations in biological samples. We analysed methadone in serum and whole blood samples using gas chromatography/mass spectrometry (GC/MS). Before analysis, we compared five solid-phase extraction (SPE) and two liquid-liquid extraction (LLE) methods and determined that SPE with Supelco LC-18 in serum yielded the best extraction efficiency. The limit of detection was 10 ng mL-1 and the limit of quantification 25 ng mL-1. Correlation coefficient was over 0.999 for the methadone calibration curve in linear range from 50 to 2000 ng mL-1. Intra and inter-day accuracy and precision of the method was satisfactory. The method was successfully applied for determining serum methadone in patients on maintenance therapy.

  10. Occurrence of gastrointestinal parasites in Georgia cattle.

    Science.gov (United States)

    Ciordia, H

    1975-04-01

    A survey of gastrointestinal nematodes in Georgia cattle was conducted from 1968 through 1973 from actual worm counts from viscera of 145 slaughtered beef cattle or from egg counts made from fecal samples from 3,273 beef and 100 dairy cattle. Beef cattle were grouped as calves, yearlings, and cows. Dairy cattle were grouped as calves, replacement heifers, and cows. Results of fecal examinations positive; 99.5, 99.3, 99.6, and 41.2% of the calves, yearlings, drylot calves, and cows, respectively, were infected with nematodes. Seventy-four percent of the eggs counted were of the Cooperia-Ostertagia - Trichostrongylus complex (C-O-T) and 21% were of the Haemonchus-Oesophagostomum complex (H-O). Sixty-nine percent of the infective larvae recovered from coprocultures made from composite samples were of Ostertagia ostertagi. Moniezia (tapeworm) eggs were seen in 16% of all the samples, more predominantly in the calves (24%) and less in the cows (smaller than 1%). Oocysts of Eimeria spp were detected in 61% of the cattle, more predominantly in younger cattle. All cattle necropsied were infected with O ostertagi. Other nematodes present, in order of decreasing frequency, were Trichostrongylus axei (97%), Haemonchus placei (66%), Cooperia punctata (63%), Cooperia oncophora (61%), Oesophagostomum radiatum (61%), Bunostomum phlebotomum (55%), Trichostrongylus colubriformis (36%), Cooperia pectinata (28%), Trichuris spp (17%), Nematodirus spathiger (9%), and Capillaria bovis (4%). Tapeworms were also recovered from 19% of the cattle at necropsy. Seventy-eight percent of the dairy cattle were positive; 98% of the calves, 80% of the heifers, and 58% of the cows. Cooperia-Ostertagia-Trichostrongylus eggs were detected in 78% and H-O eggs in 38% of the cattle. Eimeria oocysts were detected in 71% of all the dairy cattle, and Moniezia eggs were in 10% of the calves and 7% of the heifers.

  11. The comparative utility of oral swabs and probang samples for detection of foot-and-mouth disease virus infection in cattle and pigs

    DEFF Research Database (Denmark)

    Stenfeldt, Anna Carolina; Lohse, Louise; Belsham, Graham

    2013-01-01

    Foot-and-mouth disease virus (FMDV) RNA was measured using quantitative reverse transcription-PCR (qRT-PCR) assays in oralswab and probangsamples collected from cattle and pigs during experimental infections with serotype O FMDV. During acute infection, FMDV RNA was measurable in oralswabs as wel...

  12. A Schistosoma haematobium-specific real-time PCR for diagnosis of urogenital schistosomiasis in serum samples of international travelers and migrants.

    Directory of Open Access Journals (Sweden)

    Lieselotte Cnops

    Full Text Available BACKGROUND: Diagnosis of urogenital schistosomiasis by microscopy and serological tests may be elusive in travelers due to low egg load and the absence of seroconversion upon arrival. There is need for a more sensitive diagnostic test. Therefore, we developed a real-time PCR targeting the Schistosoma haematobium-specific Dra1 sequence. METHODOLOGY/PRINCIPAL FINDINGS: The PCR was evaluated on urine (n = 111, stool (n = 84 and serum samples (n = 135, and one biopsy from travelers and migrants with confirmed or suspected schistosomiasis. PCR revealed a positive result in 7/7 urine samples, 11/11 stool samples and 1/1 biopsy containing S. haematobium eggs as demonstrated by microscopy and in 22/23 serum samples from patients with a parasitological confirmed S. haematobium infection. S. haematobium DNA was additionally detected by PCR in 7 urine, 3 stool and 5 serum samples of patients suspected of having schistosomiasis without egg excretion in urine and feces. None of these suspected patients demonstrated other parasitic infections except one with Blastocystis hominis and Entamoeba cyst in a fecal sample. The PCR was negative in all stool samples containing S. mansoni eggs (n = 21 and in all serum samples of patients with a microscopically confirmed S. mansoni (n = 22, Ascaris lumbricoides (n = 1, Ancylostomidae (n = 1, Strongyloides stercoralis (n = 1 or Trichuris trichuria infection (n = 1. The PCR demonstrated a high specificity, reproducibility and analytical sensitivity (0.5 eggs per gram of feces. CONCLUSION/SIGNIFICANCE: The real-time PCR targeting the Dra1 sequence for S. haematobium-specific detection in urine, feces, and particularly serum, is a promising tool to confirm the diagnosis, also during the acute phase of urogenital schistosomiasis.

  13. Serological survey for rabies in serum samples from vampire bats (Desmodus rotundus in Botucatu region, SP, Brazil

    Directory of Open Access Journals (Sweden)

    H Langoni

    2008-01-01

    Full Text Available The chiropterans constitute 25% of the world's mammal fauna. Due to the destruction of their natural ecosystem, the vampire bats have moved from nature to artificial roosts closer to man and domestic animals. This phenomenon has happened particularly in rural areas. Rabies is a viral anthropozoonosis, 100% lethal, and vampire bats (Desmodus rotundus represent an important role in its epidemiology. D. rotundus were captured at night with mesh nets in partnership with the Botucatu Defense Office and sent to the Zoonosis Diagnostic Service, at the School of Veterinary Medicine and Animal Husbandry, UNESP. Serum samples from 204 bats were analyzed by enzyme-linked immunosorbent assay (ELISA and fluorescent antibody viral neutralization test (FAVN for rabies antibody detection. The results showed 7.4% of sera with titers higher or equal to 0.5 U for rabies antibodies, which demonstrated viral flow circulation among the studied region. Data suggest a need for constant monitoring accomplished by epidemiological and sanitary measures.

  14. Antinuclear, Cytoskeletal, Antineuronal Antibodies in the Serum Samples of Children with Tic Disorders and Obsessive Compulsive Disorders

    Directory of Open Access Journals (Sweden)

    Işık Görker

    2011-11-01

    Full Text Available streptococcus infections in the development of tic and obsessive compulsive disorders (OCD is controversial. The autoimmune hypothesis states that during infection, formation of autoantibodies leads to an autoimmune disorder, which in turn results in movement disorders, tic disorders and/or OCD. In order to test this hypothesis, we assayed these antibodies in children and adolescents diagnosed with tic disorders and/or OCD.Material and Methods: Children and adolescents who were diagnosed with either tic disorders or OCD according to DSM-IV criteria (n=28, were compared with healthy controls (n=15 having similar age and gender characteristics. Regardless of a streptococcus infection history, serum samples of all patients and controls underwent antinuclear, cytoskeletal, and antineuronal antibody assay using indirect immunofluorescence.Results: The rates of antinuclear antibody positivity were 21% and 20% in the patient and control groups respectively (p>0.05. Antineuronal antibody was positive in 2 (7% of 28 patients versus in 1 (6% of 15 controls (p>0.05.Conclusion: These results suggest that such antibodies may not be involved in the pathogenesis of tic disorders/OCD.

  15. Large volume sample stacking for rapid and sensitive determination of antidiabetic drug metformin in human urine and serum by capillary electrophoresis with contactless conductivity detection.

    Science.gov (United States)

    Tůma, Petr

    2014-06-06

    Two CE methods with contactless conductivity detection have been developed for determining the oral antidiabetic drug metformin in human urine and blood. The determination of metformin is performed on a separation capillary with an effective length of 14 cm, using a maximum voltage of 30 kV and with a small injection of 50-fold diluted urine into the capillary. Under these conditions, the migration time of metformin is 35s and the LOD is 0.3 μM. Large-volume sample stacking was used to determine low metformin levels in serum. The injection of a sample of serum deproteinized with acetonitrile was 10 times greater compared to the injected amount of urine. This enabled reduction of the LOD to 0.03 μM and the metformin migration time equalled 86 s. The undesirable solvent from sample zone was forced out of the capillary to ensure rapidity and good repeatability of the determination. The RSD values for the migration time are 0.1% for urine and 0.7% for serum; RSD for the peak areas equalled 1.4% for urine and 2.6% for serum. The developed CE technique was tested on performance of routine analyses of metformin in the urine and serum of patients suffering from type II diabetes mellitus. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Glyco-centric lectin magnetic bead array (LeMBA − proteomics dataset of human serum samples from healthy, Barrett׳s esophagus and esophageal adenocarcinoma individuals

    Directory of Open Access Journals (Sweden)

    Alok K. Shah

    2016-06-01

    Full Text Available This data article describes serum glycoprotein biomarker discovery and qualification datasets generated using lectin magnetic bead array (LeMBA – mass spectrometry techniques, “Serum glycoprotein biomarker discovery and qualification pipeline reveals novel diagnostic biomarker candidates for esophageal adenocarcinoma” [1]. Serum samples collected from healthy, metaplastic Barrett׳s esophagus (BE and esophageal adenocarcinoma (EAC individuals were profiled for glycoprotein subsets via differential lectin binding. The biomarker discovery proteomics dataset consisting of 20 individual lectin pull-downs for 29 serum samples with a spiked-in internal standard chicken ovalbumin protein has been deposited in the PRIDE partner repository of the ProteomeXchange Consortium with the data set identifier PRIDE: http://www.ebi.ac.uk/pride/archive/projects/PXD002442. Annotated MS/MS spectra for the peptide identifications can be viewed using MS-Viewer (〈http://prospector2.ucsf.edu/prospector/cgi-bin/msform.cgi?form=msviewer〉 using search key “jn7qafftux”. The qualification dataset contained 6-lectin pulldown-coupled multiple reaction monitoring-mass spectrometry (MRM-MS data for 41 protein candidates, from 60 serum samples. This dataset is available as a supplemental files with the original publication [1].

  17. Glyco-centric lectin magnetic bead array (LeMBA) - proteomics dataset of human serum samples from healthy, Barrett׳s esophagus and esophageal adenocarcinoma individuals.

    Science.gov (United States)

    Shah, Alok K; Lê Cao, Kim-Anh; Choi, Eunju; Chen, David; Gautier, Benoît; Nancarrow, Derek; Whiteman, David C; Baker, Peter R; Clauser, Karl R; Chalkley, Robert J; Saunders, Nicholas A; Barbour, Andrew P; Joshi, Virendra; Hill, Michelle M

    2016-06-01

    This data article describes serum glycoprotein biomarker discovery and qualification datasets generated using lectin magnetic bead array (LeMBA) - mass spectrometry techniques, "Serum glycoprotein biomarker discovery and qualification pipeline reveals novel diagnostic biomarker candidates for esophageal adenocarcinoma" [1]. Serum samples collected from healthy, metaplastic Barrett׳s esophagus (BE) and esophageal adenocarcinoma (EAC) individuals were profiled for glycoprotein subsets via differential lectin binding. The biomarker discovery proteomics dataset consisting of 20 individual lectin pull-downs for 29 serum samples with a spiked-in internal standard chicken ovalbumin protein has been deposited in the PRIDE partner repository of the ProteomeXchange Consortium with the data set identifier PRIDE: PXD002442. Annotated MS/MS spectra for the peptide identifications can be viewed using MS-Viewer (〈http://prospector2.ucsf.edu/prospector/cgi-bin/msform.cgi?form=msviewer〉) using search key "jn7qafftux". The qualification dataset contained 6-lectin pulldown-coupled multiple reaction monitoring-mass spectrometry (MRM-MS) data for 41 protein candidates, from 60 serum samples. This dataset is available as a supplemental files with the original publication [1].

  18. Determination of novel brominated flame retardants and polybrominated diphenyl ethers in serum using gas chromatography-mass spectrometry with two simplified sample preparation procedures.

    Science.gov (United States)

    Gao, Le; Li, Jian; Wu, Yandan; Yu, Miaohao; Chen, Tian; Shi, Zhixiong; Zhou, Xianqing; Sun, Zhiwei

    2016-11-01

    Two simple and efficient pretreatment procedures have been developed for the simultaneous extraction and cleanup of six novel brominated flame retardants (NBFRs) and eight common polybrominated diphenyl ethers (PBDEs) in human serum. The first sample pretreatment procedure was a quick, easy, cheap, effective, rugged, and safe (QuEChERS)-based approach. An acetone/hexane mixture was employed to isolate the lipid and analytes from the serum with a combination of MgSO4 and NaCl, followed by a dispersive solid-phase extraction (d-SPE) step using C18 particles as a sorbent. The second sample pretreatment procedure was based on solid-phase extraction. The sample extraction and cleanup were conducted directly on an Oasis HLB SPE column using 5 % aqueous isopropanol, concentrated sulfuric acid, and 10 % aqueous methanol, followed by elution with dichloromethane. The NBFRs and PBDEs were then detected using gas chromatography-negative chemical ionization mass spectrometry (GC-NCI MS). The methods were assessed for repeatability, accuracy, selectivity, limits of detection (LODs), and linearity. The results of spike recovery experiments in fetal bovine serum showed that average recoveries ranged from 77.9 % to 128.8 % with relative standard deviations (RSDs) from 0.73 % to 12.37 % for most of the analytes. The LODs for the analytes in fetal bovine serum ranged from 0.3 to 50.8 pg/mL except for decabromodiphenyl ethane. The proposed method was successfully applied to the determination of the 14 brominated flame retardants in human serum. The two pretreatment procedures described here are simple, accurate, and precise, and are suitable for the routine analysis of human serum. Graphical Abstract Workflow of a QuEChERS-based approach (top) and an SPE-based approach (bottom) for the detection of PBDEs and NBFRs in serum.

  19. Fabrication of an electrochemical nanoaptasensor based on AuNPs for ultrasensitive determination of cocaine in serum sample.

    Science.gov (United States)

    Roushani, Mahmoud; Shahdost-Fard, Faezeh

    2016-04-01

    Herein we describe an ultrasensitive electrochemical nanoaptasensor for the detection of one of the most dangerous narcotic drugs available, cocaine. The nanoaptasensor was constructed by the covalent attachment of a 5'-NH2-3'-gold nanoparticles terminated aptamer on the surface of a glassy carbon electrode which was deposited with gold nanoparticles (AuNPs/GCE). It is worth noting that the interaction of the cysteamine stable self-assembled monolayer on the AuNPs/GCE surface and the covalent attachment of terephthalaldehyde via amide coupling with the amine groups in the cysteamine and aptamer, respectively, resulted in the covalent attachment of the aptamer to AuNPs/GCE. The presence of gold nanoparticles both on surface of the glassy carbon electrode and in the end of the aptamer, can provide advantages such as increase of active surface area, high acceleration of the electron transfer and improved electrochemical signal, respectively. The decrease in the peak current of [Fe(CN)6](3-/4-) as the probe redox with increase of cocaine concentration, in differential pulse voltammetry as the measuring technique, from 5 pM up to 5 nM was linear and an unprecedented detection limit of 0.5pM was yielded. Furthermore, the effect of some common analgesic drugs as the potential interferents were investigated and also, to evaluate practical application of the proposed nanoaptasensor human blood serum sample as a real sample was used. Simple preparation, low operation cost, speed and validity are the decisive factors of this method motivating its application to biosensing investigation. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Determine the prevalence of Brucella spp. and Leptospira spp. in blood samples by multiplex polymerase chain reaction collected from cattle, sheep and goats in herds located in provinces of Iran

    Directory of Open Access Journals (Sweden)

    Faham Khamesipour

    2014-05-01

    Full Text Available Leptospirosis and brucellosis are common zoonosis that affect many species of mammals mostly causing economical losses. Further, very important fact is huge danger for human and animal health around the world. The purpose of the study is to determine the prevalence of Brucella spp. and Leptospira spp. using multiplex polymerase chain reaction (mPCR method, in blood samples collected from cattle, sheep and goats. In this study, a total number of 250 blood samples (5 cc of blood with ethilen diamin tetra asetic acid were collected randomly from 100 cattle, 80 sheep and 70 goats located on 6 herds in Chaharmahal Va Bakhtiari and Esfahan provinces, Iran. After DNA extraction and setting of mPCR for Brucella spp. and Leptospira spp. mPCR products were screened. The DNA of these microorganisms was detected by multiplex PCR from 31 and 21 out of 100 cattle, respectively. Four of 70 goat’s blood samples from goat breeding farms were positive for Leptospira spp. and 11 were positive for Brucella spp. Out of 80 sheep blood samples 23 were positive for Brucella spp. and 14 for Leptospira spp. The results of the present study show ruminant as an important reservoir for transmission of these zoonotic diseases to humans in Iran. mPCR has the ability to concurrently detect both Brucella and Leptospira species from blood samples of ruminants. The convenience and the possibility of detection of both bacteria at a time, strongly support the use of this mPCR for routine diagnostics.

  1. Chromosome analysis of arsenic affected cattle

    Directory of Open Access Journals (Sweden)

    S. Shekhar

    2014-10-01

    Full Text Available Aim: The aim was to study the chromosome analysis of arsenic affected cattle. Materials and Methods: 27 female cattle (21 arsenic affected and 6 normal were selected for cytogenetical study. The blood samples were collected, incubated, and cultured using appropriate media and specific methods. The samples were analyzed for chromosome number and morphology, relative length of the chromosome, arm ratio, and centromere index of X chromosome and chromosomal abnormalities in arsenic affected cattle to that of normal ones. Results: The diploid number of metaphase chromosomes in arsenic affected cattle as well as in normal cattle were all 2n=60, 58 being autosomes and 2 being sex chromosomes. From the centromeric position, karyotyping studies revealed that all the 29 pair of autosomes was found to be acrocentric or telocentric, and the sex chromosomes (XX were submetacentric in both normal and arsenic affected cattle. The relative length of all the autosome pairs and sex chrosomosome pair was found to be higher in normal than that of arsenic affected cattle. The mean arm ratio of X-chromosome was higher in normal than that of arsenic affected cattle, but it is reverse in case of centromere index value of X-chromosome. There was no significant difference of arm ratio and centromere index of X-chromosomes between arsenic affected and normal cattle. No chromosomal abnormalities were found in arsenic affected cattle. Conclusion: The chromosome analysis of arsenic affected cattle in West Bengal reported for the first time in this present study which may serve as a guideline for future studies in other species. These reference values will also help in comparison of cytological studies of arsenic affected cattle to that of various toxicants.

  2. Evidence of Cryptococcosis in cattle in Zaria Kaduna state, Nigeria

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    Emmanuella N. Akange

    2013-04-01

    Full Text Available Aim: Cryptococcosis is azoonotic infection caused by fungal of the Cryptococcus neoformans complex comprising of C. neoformans and C. gattii.The disease affects humans and animals worldwide causing morbidity and mortality. This work was carried out to determine the occurrence of cryptococcal antigens and factors associated with presence of antigens in cattle in Zaria, Nigeria. Materials and Methods: Three hundred and ninety (390 serum samples from cattle of various ages were collected from 11 farms in Zaria, Nigeria. The samples were analysed using alatex agglutination test and lateral flow assay kit which detectsthe polysaccharide capsular antigens of Cryptococcus species. Results:Out of the 390 samples tested 28 (7.17% were found to be positive using the latex agglutination test while only of these 22 (5.64% were positive using the lateral flow assay. There was a strong correlation (r=0.939, p=0.0002 between the results of the latex agglutination test and the lateral flow assay. There was no statistically significant difference (p>0.005 in positivity for cryptococcal antigens between sex, age and sex, though, there was a statistically significant difference (p<0.05 in positivity between management systems i.e. semi-intensive and intensive farming systems. Conclusions: The epidemiological value of this report lies in its demonstration that the risk of cattle and humans infection with cryptococcosis exist in farms in Zaria. The presence of this pathogen among these cattle poses an economic threat to the livestock industry due to the mastitis it causes. It also poses a significant public health threat because of its zoonotic nature and the increasing population of immunocompromised individuals. Large scale studies to determine specific risk factors and the role of the environment and experimental studies to determine what governs the transition from nasal colonisation to infection are recommended. [Vet World 2013; 6(2.000: 64-67

  3. Epidemiological survey of Crimean Congo hemorrhagic fever virus in cattle in East Darfur State, Sudan.

    Science.gov (United States)

    Ibrahim, Alaa M; Adam, Ibrahim A; Osman, Badreldin T; Aradaib, Imadeldin E

    2015-06-01

    Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne disease caused by CCHF virus (CCHFV) of the genus Nairovirus in the family Bunyaviridae. CCHFV causes subclinical infection in domestic livestock and an often fatal hemorrhagic illness in humans, with approximately 30% mortality rates. In the present study, a cross-sectional serosurvey was conducted in a total of 282 randomly selected cattle from five localities in East Darfur State, Sudan. The exposure status to CCHF was determined using enzyme-linked immunosorbent assay (ELISA) for detection of CCHFV-specific IgG antibodies in cattle serum samples. The CCHFV-specific IgG antibodies were detected in 54 out of 282 animals, accounting for a 19.14% prevalence rate. Older cattle (>2 years of age) were approximately five times more likely to be infected with the virus (OR=4.90, CI=1.28-18.98, p-value=0.02). Heavily tick-infested cattle (ticks all over the body) were at 11 times higher at risk compared to tick-free animals (OR=11.11, CI=2.86-43.25, p-value=0.01). Grazing system is another factor affecting CCHF, where cattle grazing on open system were 27 times more at risk compared to other grazing systems (OR=27.22, CI=7.46-99.24, p-value=0.001). There was an association between localities and CCHF cattle (OR=0.24, CI=0.07-0.83, p-value=0.02). This study confirms the exposure of cattle to CCHF in East Darfur and identifies potential risk factors associated with the disease. Further epidemiological studies and improved surveillance are urgently needed to prevent a possible outbreak of CCHF among humans in the Darfur region of Sudan. Copyright © 2015 Elsevier GmbH. All rights reserved.

  4. Matrix-assisted laser desorption/ionization mass spectrometry for quantitative determination of β-blocker drugs in one-drop of human serum sample.

    Science.gov (United States)

    Shrivas, Kamlesh; Patel, Devesh Kumar

    2011-01-01

    Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has been applied for the quantitative determination of β-blocker drugs in one-drop of human serum samples using drop-to-drop solvent microextraction (DDSME) as a preconcentrating probe. The optimum experimental conditions for β-blocker drugs were investigated and 1.8 μL volume of toluene for 10 min extraction time with the 5% addition of NaCl under pH 11.0 was found to be the best conditions for the separation and preconcentration of drugs from 30 μL of serum sample from a patient with high blood pressure. The optimized methodologies for DDSME/MALDI-MS analyses exhibited a good linearity with intra- and inter day precision value of 8.5-10.5% and 9.4-12.6%, respectively. The proposed DDSME/MALDI-MS offers a very simple, rapid and low-cost technique for the determination of β-blocker drugs in one drop of serum sample. The reported method has been successfully applied for the determination of propranolol and nadolol in small volume of serum sample from patient suffering from high blood pressure. In future, this technique could be applied for pharmacokinetic and clinical studies. Copyright © 2010 Elsevier B.V. All rights reserved.

  5. Automated column liquid chromatographic determination of amoxicillin and cefadroxil in bovine serum and muscle tissue using on-line dialysis for sample preparation

    NARCIS (Netherlands)

    Snippe, N; van de Merbel, N C; Ruiter, F P; Steijger, O M; Lingeman, H; Brinkman, U A

    1994-01-01

    A fully automated method is described for the determination of amoxicillin and cefadroxil in bovine serum and muscle tissue. The method is based on the on-line combination of dialysis and solid-phase extraction for sample preparation, and column liquid chromatography with ultraviolet detection. In

  6. Comparing identified and statistically significant lipids and polar metabolites in 15-year old serum and dried blood spot samples for longitudinal studies.

    Science.gov (United States)

    Kyle, Jennifer E; Casey, Cameron P; Stratton, Kelly G; Zink, Erika M; Kim, Young-Mo; Zheng, Xueyun; Monroe, Matthew E; Weitz, Karl K; Bloodsworth, Kent J; Orton, Daniel J; Ibrahim, Yehia M; Moore, Ronald J; Lee, Christine G; Pedersen, Catherine; Orwoll, Eric; Smith, Richard D; Burnum-Johnson, Kristin E; Baker, Erin S

    2017-03-15

    The use of dried blood spots (DBS) has many advantages over traditional plasma and serum samples such as the smaller blood volume required, storage at room temperature, and ability to sample in remote locations. However, understanding the robustness of different analytes in DBS samples is essential, especially in older samples collected for longitudinal studies. Here we analyzed the stability of polar metabolites and lipids in DBS samples collected in 2000-2001 and stored at room temperature. The identified and statistically significant molecules were then compared to matched serum samples stored at -80°C to determine if the DBS samples could be effectively used in a longitudinal study following metabolic disease. A total of 400 polar metabolites and lipids were identified in the serum and DBS samples using gas chromatograph/mass spectrometry (GC/MS), liquid chromatography (LC)/MS, and LC/ion mobility spectrometry-MS (LC/IMS-MS). The identified polar metabolites overlapped well between the sample types, though only one statistically significant metabolite was conserved in a case-control study of older diabetic males with low amounts of high-density lipoproteins and high body mass indices, triacylglycerides and glucose levels when compared to non-diabetic patients with normal levels, indicating that degradation in the DBS samples affects polar metabolite quantitation. Differences in the lipid identifications indicated that some oxidation occurs in the DBS samples. However, 36 statistically significant lipids correlated in both sample types. The difference in the number of statistically significant polar metabolites and lipids indicated that the lipids did not degrade to as great of a degree as the polar metabolites in the DBS samples and lipid quantitation was still possible. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  7. Serum Ghrelin Is Associated with Verbal Learning and Adiposity in a Sample of Healthy, Fit Older Adults

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    David Bellar

    2013-01-01

    Full Text Available The purpose of the present investigation was to determine the relationship between serum ghrelin concentrations, adiposity, and verbal learning in a group of healthy, fit older adults. Participants were 28 healthy older adults (age: yrs, BMI: . Participants reported to the laboratory and basic anthropometric data were collected, followed by a blood draw to quantify serum ghrelin. Participants then underwent cognitive testing that included the revised Hopkins Verbal Learning Test (HVLT, as well as the Mini-Mental Status Exam (MMSE. The results of the MMSE test revealed that the volunteers were cognitively intact (MMSE . A significant correlation emerged between serum ghrelin concentrations, 2 trials of the HVLT (Trial 1: , ; Trial 2: , , and the sum of three-site skinfold analysis (. Based upon the aforementioned relationships, it appears that fasting levels of serum ghrelin are related to both verbal learning and adiposity in healthy, fit older adults.

  8. Electrochemical determination of methimazole based on the acetylene black/chitosan film electrode and its application to rat serum samples.

    Science.gov (United States)

    Yazhen, Wang

    2011-06-01

    A novel method has been developed for the determination of methimazole, which was based on the enhanced electrochemical response of methimazole at the acetylene black/chitosan composite film modified glassy carbon electrode. The electrochemical behavior of methimazole was studied at this film electrode by cyclic voltammetry and differential pulse voltammetry. The experimental results showed that methimazole exhibited a remarkable oxidation peak at 0.63V at the film electrode. Compared with the bare glassy carbon electrode, the oxidation peak current increased greatly, and the peak potential shifted negatively, which indicated that the acetylene black/chitosan film electrode had good catalysis to the electrochemical oxidation of methimazole. The enhanced oxidation current of methimazole was indebted to the nano-porus structure of the composite film and the enlarged effective electrode area. The influences of some experimental conditions on the oxidation of methimazole were tested and the calibration plot was examined. The results indicated that the differential pulse response of methimazole was linear with its concentration in the range of 1.0×10(-7) to 2.0×10(-5)mol/L with a linear coefficient of 0.998, and in the range of 4.0×10(-5) to 3.0×10(-4)mol/L with a linear coefficient of 0.993. The detection limit was 2.0×10(-8)mol/L (S/N=3). The film electrode was used to detect the content of methimazole in rat serum samples by the standard addition method with satisfactory results. Copyright © 2011 Elsevier B.V. All rights reserved.

  9. Serosurveillance and factors associated with the presence of antibodies against bluetongue virus in dairy cattle in two eco-zones of Nepal.

    Science.gov (United States)

    Gaire, T N; Karki, S; Dhakal, I P; Khanal, D R; Bowen, R A

    2016-12-01

    Cattle play an important role in the epidemiology of bluetongue (BT) by acting as reservoir hosts. However, the status of BT virus (BTV) in dairy cattle in Nepal is unknown. The objective of this study was to estimate the prevalence of BTV antibodies in dairy cattle in two eco-zones of Nepal, and to identify the factors associated with virus exposure. The authors conducted a cross-sectional serosurvey from March 2012 through February 2013 by sampling 131 dairy cattle from seven clusters (villages) in the Chitwan district in the Terai region (southern lowlands) and the Lamjung district in the Hills region (the middle part of Nepal). Of the 131 serum samples tested, 29.3% (95% confidence interval [CI]: 21.5-37.2) were positive for BTV antibodies. Herd-level seroprevalence was 45.7% (95% CI: 30.9-61.0). Bivariate analysis indicated a positive association between seroconversion to BTV and age, and an association with breed of cattle after controlling for clustering of animals within herds. Based on this model, cattle were more likely to become seropositive as they aged. Crossbred cattle were more likely to be seropositive than those of exotic breeds (odds ratio [OR] = 4.6; 95% CI: 1.5-14.1). The results indicate widespread exposure of dairy cattle to BTV in Nepal. The authors suggest that dairy cattle should be included in the surveillance plan for BTV infection in Nepal and that it is important to educate farmers about the possible impacts of this disease. © OIE (World Organisation for Animal Health), 2016.

  10. Fabrication of an electrochemical nanoaptasensor based on AuNPs for ultrasensitive determination of cocaine in serum sample

    Energy Technology Data Exchange (ETDEWEB)

    Roushani, Mahmoud, E-mail: mahmoudroushani@yahoo.com; Shahdost-fard, Faezeh

    2016-04-01

    Herein we describe an ultrasensitive electrochemical nanoaptasensor for the detection of one of the most dangerous narcotic drugs available, cocaine. The nanoaptasensor was constructed by the covalent attachment of a 5′-NH{sub 2}-3′-gold nanoparticles terminated aptamer on the surface of a glassy carbon electrode which was deposited with gold nanoparticles (AuNPs/GCE). It is worth noting that the interaction of the cysteamine stable self-assembled monolayer on the AuNPs/GCE surface and the covalent attachment of terephthalaldehyde via amide coupling with the amine groups in the cysteamine and aptamer, respectively, resulted in the covalent attachment of the aptamer to AuNPs/GCE. The presence of gold nanoparticles both on surface of the glassy carbon electrode and in the end of the aptamer, can provide advantages such as increase of active surface area, high acceleration of the electron transfer and improved electrochemical signal, respectively. The decrease in the peak current of [Fe(CN){sub 6}]{sup 3−/4−} as the probe redox with increase of cocaine concentration, in differential pulse voltammetry as the measuring technique, from 5 pM up to 5 nM was linear and an unprecedented detection limit of 0.5 pM was yielded. Furthermore, the effect of some common analgesic drugs as the potential interferents were investigated and also, to evaluate practical application of the proposed nanoaptasensor human blood serum sample as a real sample was used. Simple preparation, low operation cost, speed and validity are the decisive factors of this method motivating its application to biosensing investigation. - Highlights: • An electrochemical nanoaptasensor for the detection of cocaine is presented. • An AuNPs terminated aptamer was covalent bonded on the surface of the AuNPs/GCE. • The presence of AuNPs has many advantages and improved electrochemical signal. • Two linear ranges from 5 pM up to 5 nM and an unprecedented LOD of 0.5 pM were yielded. • It will shed

  11. Assessment of the probability of introduction of bovine tuberculosis to Danish cattle farms via imports of live cattle from abroad and immigrant workers

    DEFF Research Database (Denmark)

    Foddai, Alessandro; Nielsen, Liza Rosenbaum; Krogh, Kaspar

    2015-01-01

    Denmark has been recognized as officially free (OTF) from bovine tuberculosis (bTB) since 1980. In this study, we estimated the annual probability (PIntro) of introducing Mycobacterium bovis into the Danish cattle population, through (a) imports of cattle and (b) foreign personnel working in Danish...... cattle herds. Data from 2000 to 2013 with date, number and origin of imported live cattle were obtained from the Danish Cattle Federation. Information on immigrants working in Danish cattle herds was obtained through a questionnaire sent by email to a sample of Danish cattle farmers (N = 460). Inputs...

  12. Simultaneous determination of anthracyclines and taxanes in human serum using online sample extraction coupled to high performance liquid chromatography with UV detection.

    Science.gov (United States)

    Bermingham, Shane; O'Connor, Robert; Regan, Fiona; McMahon, Gillian P

    2010-06-01

    An online SPE-LC method that can determine both anthracyclines and taxanes simultaneously in human serum samples is reported. The entire method of extraction, separation and UV detection was achieved online by column switching between an SPE column (Biotrap 500 (20 x 4 mm)) and an analytical column (Zorbax XDB C18, 150 x 4.6 mm, 5 microm) with a 23 min total cycle time. The method is linear (r(2)>0.998) over the range of 0.5-25 microg/mL. The analytes of interest are retained on the SPE column with good recovery (84-117%), while proteins and other serum components elute to waste. This online clean-up is much faster (150 s) and less manual than traditional off-line extraction methods. Using 0.1 mL spiked serum samples, the LOQ was 0.5 microg/mL. Intra- and inter-day precision were acceptable (serum samples from patients undergoing chemotherapy with these agents.

  13. Hypovitaminosis A coupled to secondary bacterial infection in beef cattle

    Directory of Open Access Journals (Sweden)

    He Xiuyuan

    2012-11-01

    Full Text Available Abstract Background Vitamin A is essential for normal growth, development, reproduction, cell proliferation, cell differentiation, immune function and vision. Hypovitaminosis A can lead to a series of pathological damage in animals. This report describes the case of hypovitaminosis A associated with secondary complications in calves. Case presentation From February to March in 2011, 2-and 3-month old beef calves presented with decreased eyesight, apparent blindness and persistent diarrhea occurred in a cattle farm of Hubei province, China. Based on history inspection and clinical observation, we made a tentative diagnosis of hypovitaminosis A. The disease was confirmed as a congenital vitamin A deficiency by determination of the concentrations of vitamin A in serum and feed samples. Furthermore, pathological and microbiological examination showed that the disease was associated with pathogenic Escherichia coli (E. coli infection and mucosal barriers damage in intestines. The corresponding treatments were taken immediately, and the disease was finally under control for a month. Conclusions To our knowledge, this is the first report of hypovitaminosis A coupled to secondary infection of E. coli in beef cattle, advancing our knowledge of how vitamin A affects infection and immunity in animals. This study could also be contributed to scientific diagnosis and treatments of complex hypovitaminosis A in cattle.

  14. Seroprevalence of brucellosis among cattle slaughtered in three municipal abattoirs of Gombe state, Northeastern Nigeria

    Directory of Open Access Journals (Sweden)

    Saleh Mohammed Jajere

    2016-10-01

    Full Text Available Aim: A cross-sectional study was conducted to determine the seroprevalence of bovine brucellosis among cattle slaughtered at three municipal abattoirs of Gombe State, Nigeria. Materials and Methods: A total of 200 blood samples collected from slaughtered cattle of different breeds (Sokoto Gudali - 50, White Fulani - 102, Red bororo – 34, and Crossbreeds - 14, sex (males - 19 and females - 181, and from different locations (Billiri - 30, Yamaltu Deba – 50, and Gombe - 120 were screened for brucellosis using rose bengal plate test (RBPT, serum agglutination test (SAT, and microtiter agglutination test (MAT. Results: Of the 200 serum samples analyzed, 7 (3.5%, 10 (5.0% and 18 (9.0% were positive by RBPT, SAT and MAT, respectively. The results showed no statistically significant association between sex and seropositivity to bovine brucellosis. However, seropositivity of bovine brucellosis was higher in females than in males. Similarly, no statistically significant association was observed between breed and occurrence of bovine brucellosis. Moreover, the prevalence of brucellosis was higher in Sokoto Gudali as compared with the other breeds. Based on the study locations, higher seroprevalence was observed in animals screened from Billiri as compared with those from other locations (p<0.05. Conclusion: The presence of Brucella abortus antigen in the sera of slaughtered cattle in Gombe state poses a significant public health risk. Therefore, it is important to carry out further epidemiological studies on fulani herdsmen and cattle herds in the study area, in order to explore the risk factors associated with the occurrence and perpetuation of brucellosis among cattle herds, ascertain the prevalence and status of the disease among both farms and nomadic herds.

  15. Craniometrical estimation of the native Japanese Mishima cattle, using multivariate analysis.

    Science.gov (United States)

    Ogawa, Y; Daigo, M; Amasaki, H

    1989-01-01

    The present study on measurement of the skull of Mishima cattle, which has been postulated as the only pure representative breed of native Japanese cattle, was performed using craniometrical multivariate analysis. The data of the skull of Mishima cattle was compared with 17 breeds of cattle, i.e. Korean cattle (Hamhung, Pyongyang, Chinju Suwon, and Kwangju), Mongolian cattle, Hainan Tao cattle, northeastern Chinese cattle (Shuangliao, Shenyang, Tongliao, Lüta, and Chilin), Astatic Water Buffalo, Yak, Bos Banteng, American Bison, and Holstein-Friesian. The Mishima cattle was included in the group of Korean breeds, especially it was closed on the group of Pyongyang and Chinju breeds. The distance on the craniometrical multivariate analyzing co-ordinate between Mishima cattle and Hainan Tao breed of Zebu cattle was larger than the distance between Mishima cattle and Korean breeds. While result, as a above the present study was very important for the origin of "Wagyu" (native Japanese cattle). Since the northern route theory of the origin of Mishima cattle has been reported on the type of serum enzymes and hemotypes. It was suggested that the craniometrical multivariate analysis supported to the northern route theory of the origin of Mishima cattle.

  16. Tissue and serum samples of patients with papillary thyroid cancer with and without benign background demonstrate different altered expression of proteins

    Science.gov (United States)

    Abdullah, Mardiaty Iryani; Lee, Ching Chin; Mat Junit, Sarni; Ng, Khoon Leong

    2016-01-01

    Background Papillary thyroid cancer (PTC) is mainly diagnosed using fine-needle aspiration biopsy. This most common form of well-differentiated thyroid cancer occurs with or without a background of benign thyroid goiter (BTG). Methods In the present study, a gel-based proteomics analysis was performed to analyse the expression of proteins in tissue and serum samples of PTC patients with (PTCb; n = 6) and without a history of BTG (PTCa; n = 8) relative to patients with BTG (n = 20). This was followed by confirmation of the levels of proteins which showed significant altered abundances of more than two-fold difference (p AHSG). The different altered expression of tissue and serum A1AT as well as serum AHSG between PTCa and PTCb patients were also validated by ELISA. Discussion The distinctive altered abundances of the tissue and serum proteins form preliminary indications that PTCa and PTCb are two distinct cancers of the thyroid that are etiologically and mechanistically different although it is currently not possible to rule out that they may also be due other reasons such as the different stages of the malignant disease. These proteins stand to have a potential use as tissue or serum biomarkers to discriminate the three different thyroid neoplasms although this requires further validation in clinically representative populations. PMID:27672505

  17. Bluetongue virus surveillance in the Islamic Republic of Mauritania: Is serotype 26 circulating among cattle and dromedaries?

    Science.gov (United States)

    Lorusso, Alessio; Baba, Doumbia; Spedicato, Massimo; Teodori, Liana; Bonfini, Barbara; Marcacci, Maurilia; Di Provvido, Andrea; Isselmou, Katia; Marini, Valeria; Carmine, Irene; Scacchia, Massimo; Di Sabatino, Daria; Petrini, Antonio; Bezeid, Beyatt Ahmed; Savini, Giovanni

    2016-06-01

    In March 2013, EDTA-blood and serum samples were collected from 119 cattle and 159 dromedaries at the slaughterhouse of Nouakchott, the capital city of the Islamic Republic of Mauritania. Serum samples were screened for the presence of Bluetongue (BT) antibodies by competitive ELISA (cELISA). Positive samples were then tested by serum-neutralization (SN) to determine BTV serotype. RNA from blood samples was first tested by a genus-specific quantitative RT-PCR assay which is able to detect all 27 existing BTV serotypes (RT-qPCR1-27). Positive samples were further screened by a RT-qPCR assay which, instead, is able to detect the classical 24 BTV serotypes only (RT-qPCR1-24). Of the 278 serum samples tested, 177 (mean=63.7%; 95% CI: 57.9%-69.1%) resulted positive by cELISA. Of these, 69 were from cattle (mean=58.0%; 95% CI: 49.0%-66.5%) and 108 from dromedaries (mean=67.9%; 95% CI: 60.3%-74.7%). BTV-26 neutralizing antibodies were by far the most frequently found as they were detected in 146 animals with titres ranging from 1:10 to 1:80. Out of 278 blood samples, 25 (mean=9.0%; 95% CI: 6.2%-12.9%) were found positive for BTV by RT-qPCR1-27, 20 (mean=16.8%; 95% CI: 11.2%-24.6%) were from cattle and 5 (mean=3.1%; 95% CI: 1.4%-7.1%) from dromedaries. When tested by RT-qPCR1-24 the 25 BTV positive samples were negative. Unfortunately, no genetic information by molecular typing or by next generation sequencing has been obtained as for the very low levels of RNA in the blood samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Preparation of Fe3O4 nanoparticle enclosure hydroxylated multi-walled carbon nanotubes for the determination of aconitines in human serum samples.

    Science.gov (United States)

    Zhang, Hong-Fei; Shi, Yan-Ping

    2012-04-29

    A magnetic carbon nanomaterial for Fe(3)O(4) enclosure hydroxylated multi-walled carbon nanotubes (Fe(3)O(4)-EC-MWCNTs-OH) was prepared by the aggregating effect of Fe(3)O(4) nanoparticle on MWCNTs-OH, and combined with high-performance liquid chromatography (HPLC)/diode array detection (DAD) to determine the aconitines (aconitine, hypaconitine and mesaconitine) in human serum samples. Compared with other extraction modes investigated in experiment, Fe(3)O(4)-EC-MWCNTs-OH sorbents showed a good affinity to target analytes. Some important parameters that could influence extraction efficiency of aconitines, including the extraction mode, amounts of Fe(3)O(4)-EC-MWCNTs-OH, pH of sample solution, extraction time, desorption solvent and desorption time, were optimized. Under optimal conditions, the recoveries of spiked serum samples were between 98.0% and 103.0%; relative standard deviations (RSDs) ranged from 0.9% to 6.2%. The correlation coefficients varied from 0.9996 to 0.9998. The limits of detection ranged from 3.1 ng mL(-1) to 4.1 ng mL(-1) at a signal-to-noise ratio of 3. The experimental results showed that the proposed method was feasible for the analysis of aconitines in serum samples. Copyright © 2012 Elsevier B.V. All rights reserved.

  19. Detection of EBV-DNA in serum samples of an immunosuppressed child during a three years follow-up: association of clinical and PCR data with active infection.

    Science.gov (United States)

    Okay, Thelma Suely; Del Negro, Gilda Maria Barbaro; Yamamoto, Lídia; Raiz Júnior, Roberto

    2005-01-01

    Twenty-four whole blood and serum samples were drawn from an eight year-old heart transplant child during a 36 months follow-up. EBV serology was positive for VCA-IgM and IgG, and negative for EBNA-IgG at the age of five years old when the child presented with signs and symptoms suggestive of acute infectious mononucleosis. After 14 months, serological parameters were: positive VCA-IgG, EBNA-IgG and negative VCA-IgM. This serological pattern has been maintained since then even during episodes suggestive of EBV reactivation. PCR amplified a specific DNA fragment from the EBV gp220 (detection limit of 100 viral copies). All twenty-four whole blood samples yielded positive results by PCR, while 12 out of 24 serum samples were positive. We aimed at analyzing whether detection of EBV-DNA in serum samples by PCR was associated with overt disease as stated by the need of antiviral treatment and hospitalization. Statistical analysis showed agreement between the two parameters evidenced by the Kappa test (value 0.750; p Real-Time PCR or another quantitative method is not available.

  20. Tissue and serum samples of patients with papillary thyroid cancer with and without benign background demonstrate different altered expression of proteins

    Directory of Open Access Journals (Sweden)

    Mardiaty Iryani Abdullah

    2016-09-01

    Full Text Available Background Papillary thyroid cancer (PTC is mainly diagnosed using fine-needle aspiration biopsy. This most common form of well-differentiated thyroid cancer occurs with or without a background of benign thyroid goiter (BTG. Methods In the present study, a gel-based proteomics analysis was performed to analyse the expression of proteins in tissue and serum samples of PTC patients with (PTCb; n = 6 and without a history of BTG (PTCa; n = 8 relative to patients with BTG (n = 20. This was followed by confirmation of the levels of proteins which showed significant altered abundances of more than two-fold difference (p < 0.01 in the tissue and serum samples of the same subjects using ELISA. Results The data of our study showed that PTCa and PTCb distinguish themselves from BTG in the types of tissue and serum proteins of altered abundance. While higher levels of alpha-1 antitrypsin (A1AT and heat shock 70 kDa protein were associated with PTCa, lower levels of A1AT, protein disulfide isomerase and ubiquitin-conjugating enzyme E2 N seemed apparent in the PTCb. In case of the serum proteins, higher abundances of A1AT and alpha 1-beta glycoprotein were detected in PTCa, while PTCb was associated with enhanced apolipoprotein A-IV and alpha 2-HS glycoprotein (AHSG. The different altered expression of tissue and serum A1AT as well as serum AHSG between PTCa and PTCb patients were also validated by ELISA. Discussion The distinctive altered abundances of the tissue and serum proteins form preliminary indications that PTCa and PTCb are two distinct cancers of the thyroid that are etiologically and mechanistically different although it is currently not possible to rule out that they may also be due other reasons such as the different stages of the malignant disease. These proteins stand to have a potential use as tissue or serum biomarkers to discriminate the three different thyroid neoplasms although this requires further validation in clinically

  1. Utility of acute phase proteins as biomarkers of transport stress in ewes and beef cattle

    Directory of Open Access Journals (Sweden)

    Francesco Fazio

    2015-05-01

    Full Text Available The effect of transport on serum amyloid A (SAA, haptoglobin (Hp, Fibrinogen and white blood cells (WBC was evaluated in 10 ewes and 10 beef cattle. All animals were transported by road for 6 h over a distance of about 490 km with an average speed of 80 km/h. Blood samples, collected via jugular venepuncture, were obtained before and after transport as well as after 12, 24 and 48 h rest time. One-way repeated measures analysis of variance showed a statistically significant effect of sampling time on SAA, Hp, and WBC in ewes and beef cattle. Based on these results, Hp and SAA levels, together with WBC, may be useful indicators of animal health and welfare and in predicting the risk assessment in meat inspection.

  2. Reliability of Serum Metabolites over a Two-Year Period: A Targeted Metabolomic Approach in Fasting and Non-Fasting Samples from EPIC

    Science.gov (United States)

    Achaintre, David; Sacerdote, Carlotta; Vineis, Paolo; Key, Timothy J.; Onland Moret, N. Charlotte; Scalbert, Augustin; Rinaldi, Sabina; Ferrari, Pietro

    2015-01-01

    Objective Although metabolic profiles have been associated with chronic disease risk, lack of temporal stability of metabolite levels could limit their use in epidemiological investigations. The present study aims to evaluate the reliability over a two-year period of 158 metabolites and compare reliability over time in fasting and non-fasting serum samples. Methods Metabolites were measured with the AbsolueIDQp180 kit (Biocrates, Innsbruck, Austria) by mass spectrometry and included acylcarnitines, amino acids, biogenic amines, hexoses, phosphatidylcholines and sphingomyelins. Measurements were performed on repeat serum samples collected two years apart in 27 fasting men from Turin, Italy, and 39 non-fasting women from Utrecht, The Netherlands, all participating in the European Prospective Investigation into Cancer and Nutrition (EPIC) study. Reproducibility was assessed by estimating intraclass correlation coefficients (ICCs) in multivariable mixed models. Results In fasting samples, a median ICC of 0.70 was observed. ICC values were fasting samples, the median ICC was 0.54. ICC values were fasting as compared to fasting samples, with a statistically significant difference for 19–36% of acylcarnitines, phosphatidylcholines and sphingomyelins. Conclusion A single measurement per individual may be sufficient for the study of 73% and 52% of the metabolites showing ICCs >0.50 in fasting and non-fasting samples, respectively. ICCs were higher in fasting samples that are preferable to non-fasting. PMID:26274920

  3. Reliability of Serum Metabolites over a Two-Year Period: A Targeted Metabolomic Approach in Fasting and Non-Fasting Samples from EPIC.

    Science.gov (United States)

    Carayol, Marion; Licaj, Idlir; Achaintre, David; Sacerdote, Carlotta; Vineis, Paolo; Key, Timothy J; Onland Moret, N Charlotte; Scalbert, Augustin; Rinaldi, Sabina; Ferrari, Pietro

    2015-01-01

    Although metabolic profiles have been associated with chronic disease risk, lack of temporal stability of metabolite levels could limit their use in epidemiological investigations. The present study aims to evaluate the reliability over a two-year period of 158 metabolites and compare reliability over time in fasting and non-fasting serum samples. Metabolites were measured with the AbsolueIDQp180 kit (Biocrates, Innsbruck, Austria) by mass spectrometry and included acylcarnitines, amino acids, biogenic amines, hexoses, phosphatidylcholines and sphingomyelins. Measurements were performed on repeat serum samples collected two years apart in 27 fasting men from Turin, Italy, and 39 non-fasting women from Utrecht, The Netherlands, all participating in the European Prospective Investigation into Cancer and Nutrition (EPIC) study. Reproducibility was assessed by estimating intraclass correlation coefficients (ICCs) in multivariable mixed models. In fasting samples, a median ICC of 0.70 was observed. ICC values were fasting samples, the median ICC was 0.54. ICC values were fasting as compared to fasting samples, with a statistically significant difference for 19-36% of acylcarnitines, phosphatidylcholines and sphingomyelins. A single measurement per individual may be sufficient for the study of 73% and 52% of the metabolites showing ICCs >0.50 in fasting and non-fasting samples, respectively. ICCs were higher in fasting samples that are preferable to non-fasting.

  4. Untargeted metabolomic analysis of human serum samples associated with different levels of red meat consumption: A possible indicator of type 2 diabetes?

    Science.gov (United States)

    Carrizo, Daniel; Chevallier, Olivier P; Woodside, Jayne V; Brennan, Sarah F; Cantwell, Marie M; Cuskelly, Geraldine; Elliott, Christopher T

    2017-04-15

    Red meat consumption has been associated with negative health effects. A study to identify biomarkers of meat consumption was undertaken using serum samples collected from combining high resolution mass spectrometry (UPLC-QTof-MS) and chemometrics. Using orthogonal partial last-squares discriminant analysis (OPLS-DA), multivariate models were created for both modes of acquisition (ESI-/ESI+) and red meat intake classes (YES/NO). In the serum samples, a total 3280 and 3225 ions of interest were detected in positive and negative modes, respectively. Of these, 62 were found to be significantly different (pmeat intake groups. This study has shown metabolomics fingerprints have the capability to identify potential biomarkers of red meat consumption, as well as possible health risk factors (e.g., key metabolic families related to the risk of development type 2 diabetes). Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Determination of donepezil in serum samples using molecularly imprinted polymer nanoparticles followed by high-performance liquid chromatography with ultraviolet detection.

    Science.gov (United States)

    Khansari, Mehdi Rajabnia; Bikloo, Shahrzad; Shahreza, Sara

    2016-03-01

    A molecularly imprinted polymer designed for the selective extraction of donepezil from serum samples was synthesized using a noncovalent molecular imprinting approach. The molecularly imprinted polymer was evaluated chromatographically and then its affinity for donepezil was confirmed by solid-phase extraction. The optimal conditions for solid-phase extraction were provided by cartridge conditioning using acidified water purified from a Milli-Q system, sample loading under basic aqueous conditions, clean-up using acetonitrile, and elution with methanol/tetrahydrofuran. Desirable molecular recognition properties of the molecularly imprinted polymer led to good donepezil recoveries (90-102%). The data indicated that the imprinted polymer has a perfect selectivity and affinity for donepezil and could be used for selective extraction and analysis of donepezil in human serum. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Bodily fluid analysis of non-serum samples using point-of-care testing with iSTAT and Piccolo analyzers versus a fixed hospital chemistry analytical platform.

    Science.gov (United States)

    Londeree, William; Davis, Konrad; Helman, Donald; Abadie, Jude

    2014-09-01

    Forward deployed military medical units can provide sophisticated medical care with limited resources. Point-of-Care Testing (POCT) may facilitate care and expedite diagnosis. This study assessed the accuracy of results for POCT for non-serum samples (pleural, peritoneal, and cerebrospinal fluid) using iSTAT and Piccolo hand-held devices compared with results obtained using a hospital chemistry analyzer. Pleural, peritoneal, and cerebrospinal fluids obtained during routine care were simultaneously analyzed on a Vitros 5600 automated clinical chemistry hospital analyzer, iSTAT, and Piccolo POCT devices. POCT results were highly correlated with the Vitros 5600 for pleural fluid LDH, glucose, and triglycerides (TG); for peritoneal fluid bilirubin, TG, glucose, albumin, and protein; and glucose for cerebrospinal fluid. POCT results for non-serum samples from pleural, peritoneal, and cerebrospinal fluid correlate with standard hospital chemistry analysis. The results of this study demonstrate potential for possible new diagnostic roles for POCT in resource-limited environments.

  7. Simultaneous UHPLC-UV analysis of hydroxychloroquine, minocycline and doxycycline from serum samples for the therapeutic drug monitoring of Q fever and Whipple's disease.

    Science.gov (United States)

    Armstrong, Nicholas; Richez, Magali; Raoult, Didier; Chabriere, Eric

    2017-08-15

    A fast UHPLC-UV method was developed for the simultaneous analysis of Hydroxychloroquine, Minocycline and Doxycycline drugs from 100μL of human serum samples. Serum samples were extracted by liquid-liquid extraction and injected into a phenyl hexyl reverse phase column. Compounds were separated using a mobile phase linear gradient and monitored by UV detection at 343nm. Chloroquine and Oxytetracycline were used as internal standards. Lower and upper limits of quantifications, as well as the other levels of calibration, were validated with acceptable accuracy (<15% deviation) and precision (<15% coefficient of variation) according to the European Medicines Agency guidelines. This new method enables cost and time reduction and was considered suitable for the clinical laboratory. It is the first published assay for the therapeutic drug monitoring of patients diagnosed with Q fever or Whipple's disease. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Precise simultaneous quantification of methadone and cocaine in rat serum and brain tissue samples following their successive i.p. administration.

    Science.gov (United States)

    Nakhla, David S; Hussein, Lobna A; Magdy, N; Abdallah, Inas A; Hassan, Hazem E

    2017-03-24

    A sensitive high-performance liquid chromatography (HPLC) assay with dual UV detection has been developed and validated for the simultaneous quantification of methadone and cocaine in rat serum and brain tissue samples. Liquid-liquid extraction using hexanes was applied for samples extraction with Levo-Tetrahydropalmatine (L-THP) as the internal standard. Chromatographic separation of the analytes was achieved on a reversed-phase Waters Symmetry® C18 column (150mm×4.6mm, 5μm). A gradient elution was employed with a mobile phase consisting of 5mM potassium phosphate containing 0.1% triethylamine (pH=6.5) (A) and acetonitrile (B) with a flow rate of 1mL/min. UV detection was employed at 215nm and 235nm for the determination of methadone and cocaine, respectively. The calibration curves were linear over the range of 0.05-10μg/mL for both methadone and cocaine. The assay was validated according to FDA guidelines for bioanalytical method validation and results were satisfactory and met FDA criteria. Inter-day accuracy values of serum and brain samples ranged from 96.97 to 105.59% while intra-day accuracy values ranged from 91.49 to 111.92%. Stability assays showed that both methadone and cocaine were stable during sample storage, preparation, and analytical procedures. The method was successfully used to analyze biological samples obtained from a drug- drug interaction pharmacokinetics (PK) study conducted in rats to investigate the effect of methadone on cocaine PK. Our method not only can be used for bioanalysis of samples obtained from rats but also can potentially be applied to human biological serum samples to monitor compliance to methadone maintenance therapy (MMT) and to detect possible cocaine-methadone co-abuse. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Reliability of Serum Metabolites over a Two-Year Period : A Targeted Metabolomic Approach in Fasting and Non-Fasting Samples from EPIC

    OpenAIRE

    Marion Carayol; Idlir Licaj; David Achaintre; Carlotta Sacerdote; Paolo Vineis; Key, Timothy J.; N Charlotte Onland Moret; Augustin Scalbert; Sabina Rinaldi; Pietro Ferrari

    2015-01-01

    Objective Although metabolic profiles have been associated with chronic disease risk, lack of temporal stability of metabolite levels could limit their use in epidemiological investigations. The present study aims to evaluate the reliability over a two-year period of 158 metabolites and compare reliability over time in fasting and non-fasting serum samples. Methods Metabolites were measured with the AbsolueIDQp180 kit (Biocrates, Innsbruck, Austria) by mass spectrometry a...

  10. Neospora caninum: Seroprevalence in beef cattle in the mountainous region of Santa Catarina, Brazil

    Directory of Open Access Journals (Sweden)

    Mayckon Antonio Cardoso Padilha

    2017-03-01

    Full Text Available The distribution of Neospora caninum, an important agent of reproductive disorders in cattle, has been frequently reported in dairy cattle in Brazil and worldwide. Records of infection are less frequent in beef cattle. To determine the seroprevalence of bovine neosporosis, 507 beef cattle blood samples were collected from January 2013 to September 2015, from 16 municipalities of the Associação dos Municípios da Região Serrana (AMURES. Samples were tested for the presence of IgG antibodies ( ? 1:100 against N. caninum by indirect immunofluorescence antibody test (IFAT. Information about sex, age, and origin of the bovines were obtained from the Brazilian Bovine and Bubaline Identification and Certification System (SISBOV, and were tabulated for statistical analysis (Chi-square and Fisher Exact test, P ? 0.05. Of the 507 serum samples analyzed, 70 (13.81% contained antibodies against N. caninum with titers of 1:100 (16, 1:200 (22, 1:400 (17, 1:800 (nine, 1:1600 (four and 1:3200 (two. Positive specimens were collected from 13 of the 16 municipalities and across all age groups. Of the positive samples, there were no statistical differences between males and females (32.86% vs. 67.14%, P= 0.1072, age groups (P= 0.4116, or municipalities (P= 0.6838. While not statistically significant (P>0.05, higher seroprevalence was observed among older females. Although seroprevalence was relatively low, results indicate that infection by N. caninum in beef cattle is widespread in the studied region.

  11. Serum sickness

    Science.gov (United States)

    Drug allergy - serum sickness; Allergic reaction - serum sickness; Allergy - serum sickness ... symptoms of serum sickness. Certain medicines (such as penicillin, cefaclor, and sulfa) can cause a similar reaction. ...

  12. Direct comparison of galactomannan performance in concurrent serum and bronchoalveolar lavage samples in immunocompromised patients at risk for invasive pulmonary aspergillosis.

    Science.gov (United States)

    Boch, Tobias; Buchheidt, Dieter; Spiess, Birgit; Miethke, Thomas; Hofmann, Wolf-Karsten; Reinwald, Mark

    2016-02-01

    Invasive pulmonary aspergillosis (IPA) is a life-threatening infection mainly affecting immunocompromised patients. Early diagnosis is critical, but the diagnostic gold standard (histopathology and culture) is time consuming and cannot offer early confirmation of IPA. Fungal biomarkers like galactomannan (GM) are a promising extension to the diagnostic repertoire. However, it still remains under discussion if biomarker analysis from the site of the infection is superior to testing blood samples. We retrospectively evaluated the diagnostic performance of concurrent serum GM and bronchoalveolar lavage (BAL) GM (obtained within 24 h) of immunocompromised patients at high risk of IPA. Twenty-six proven/probable patients and eight patients with no IPA according to the EORTC/MSG 2008 criteria were included in this study. Sensitivity, specificity, positive predictive value, negative predictive value and diagnostic odds ratio were for BAL GM: 85%, 88%, 96%, 64% and 38.5, and for serum GM: 23%, 88%, 88%, 26% and 2.1 respectively. BAL GM proved to be significantly more sensitive for the detection of IPA compared to same-day serum GM in patients at high risk of IPA (P superior to serum GM implying that diagnostic efforts should focus on specimens from the site of infection. © 2015 Blackwell Verlag GmbH.

  13. Effect of ambient temperature on viral replication and serum antibody titers following administration of a commercial intranasal modified-live infectious bovine rhinotracheitis-parainfluenza-3 virus vaccine to beef cattle housed in high- and moderate-ambient temperature environments.

    Science.gov (United States)

    Grissett, Gretchen P; White, Brad J; Anderson, David E; Larson, Robert E; Miesner, Matt D

    2014-12-01

    To evaluate the effect of ambient temperature on viral replication and serum antibody titers following administration of an intranasal modified-live infectious bovine rhinotracheitis (IBR)-parainfluenza-3 (PI3) virus vaccine to beef calves housed in high- (> 32°C) and moderate- (21°C) ambient temperature environments. 28 calves (mean weight, 206.8 kg). Calves were randomly allocated to 4 treatment groups (housed outdoors during high ambient temperature with [HAT; n = 10] or without [HAC; 4] vaccination or housed indoors in a moderate ambient temperature with [MAT; 10] or without [MAC; 4] vaccination). Rectal and nasal mucosal temperatures were recorded every 2 hours from 8 AM to 8 PM on days 0 (vaccination) and 1. Nasal swab specimens were obtained on days 0 through 7 for virus isolation. Serum samples were collected on days 0, 7, 14, and 28 for determination of antibody titers. Mean rectal temperature did not differ among the treatment groups. Mean nasal temperature for the HAT group was significantly higher than that for the MAT group at 6, 24, 30, 32, and 38 hours after vaccination. Viable IBR virus was isolated from all vaccinated calves on days 1 through 6. Two weeks after vaccination, vaccinated calves had anti-IBR antibody titers that were significantly greater than those for unvaccinated calves. Mean anti-IBR antibody titers did not differ significantly between the HAT and MAT groups. Results indicated that, following vaccination with an intranasal modified-live IBR-PI3 virus vaccine, IBR viral replication and serum antibody titers did not differ significantly between calves housed in high- and moderate-ambient temperature environments.

  14. Measurements of C-reactive protein (CRP) and nerve-growth-factor (NGF) concentrations in serum and urine samples of dogs with neurologic disorders.

    Science.gov (United States)

    Kordass, Ulrike; Carlson, Regina; Stein, Veronika Maria; Tipold, Andrea

    2016-01-08

    The purpose of this study was to prove the hypothesis that C-reactive protein (CRP) and nerve growth factor (NGF) may be potential biomarkers for lower urinary tract disorders and may be able to distinguish between micturition dysfunctions of different origin in dogs with spinal cord diseases. NGF- and CRP- concentrations were measured in serum and urine samples using specific ELISA-Kits. Results in urine were standardized by urine-creatinine levels. CRP in serum was detectable in 32/76 and in urine samples in 40/76 patients. NGF could be measured in all serum and in 70/76 urine samples. Urinary CRP concentrations were significantly higher in dogs with micturition dysfunction (p = 0.0009) and in dogs with different neurological diseases (p = 0.0020) compared to the control group. However, comparing dogs with spinal cord disorders with and without associated micturition dysfunction no significant difference could be detected for NGF and CRP values in urine or serum samples. Additionally, levels did not decrease significantly, when measured at the time when the dogs regained the ability to urinate properly (urinary NGF p = 0.7962; urinary CRP p = 0.078). Urine samples with bacteria and/or leukocytes had no significant increase in urinary NGF (p = 0.1112) or CRP (p = 0.0534) concentrations, but higher CRP-levels in urine from dogs with cystitis were found compared to dogs without signs of cystitis. From these data we conclude that neither CRP nor NGF in urine or serum can be considered as reliable biomarkers for micturition disorders in dogs with spinal cord disorders in a clinical setting, but their production might be part of the pathogenesis of such disorders. Significantly higher levels of CRP could be found in the urine of dogs with micturition dysfunctions compared to control dogs. This phenomenon could potentially be explained by unspecific extrahepatic CRP production by smooth muscle cells in the dilated bladder.

  15. Survey Of Ketosis And Hypoproteinaemia In Slaughtered Cattle In ...

    African Journals Online (AJOL)

    Serum ketone and total protein concentrations of 966 cattle slaughtered at the metropolitan abattoir, Maiduguri, Nigera, were estimated durng the dry and rainy months of the year. None of the sera had a titrable ketone concentration. Serum total protein (STP) concentration of <60.0 g/L,considered as hypoproteinaemia, was ...

  16. An 8-year longitudinal sero-epidemiological study of bovine leukaemia virus (BLV) infection in dairy cattle in Turkey and analysis of risk factors associated with BLV seropositivity.

    Science.gov (United States)

    Şevik, Murat; Avcı, Oğuzhan; İnce, Ömer Barış

    2015-04-01

    Enzootic bovine leukosis (EBL) which is caused by bovine leukaemia virus (BLV) has an important economic impact on dairy herds due to reduced milk production and restrictions on livestock exports. This study was conducted to determine the BLV infection status in Central Anatolia Region of Turkey, an important milk production centre, and to examine the risk factors such as purchasing cattle, increasing cattle age, cattle breed and herd size associated with transmission of BLV infection. To estimate the rate of BLV infection, a survey for specific antibodies in 28,982 serum samples from animals belonging to 1116 different herds situated in Central Anatolia Region of Turkey were tested from January 2006 to December 2013. A generalized mixed linear model was used to evaluate the risk factors that influenced BLV seroprevalence. Antibodies against BLV were detected in 431 (2.28 %) of 18,822 Holstein and 29 (0.28 %) of 10,160 Brown Swiss cows. Among 1116 herds, 132 herds (11.82 %) had one or more positive animals. Also results of our study show that the prevalence of BLV infection increased from 2006 to 2011, and it tends to reduce with BLV control programme. Furthermore, we found positive associations between percentage of seropositive animal and increasing cattle age, herd size, cattle breed and purchased cattle. Age-specific prevalence showed that BLV prevalence increased with age. These factors should be taken into consideration for control of BLV infection.

  17. The Agersoe cattle

    DEFF Research Database (Denmark)

    Withen, K.B.; Brüniche-Olsen, A.; Pedersen, Bo Vest

    2011-01-01

    A phenotypically interesting strain of cattle existed on the small island of Agersoe, on the west coast of Zealand, Denmark, in the beginning of the last decade. The cattle share a great resemblance to the extinct Danish breed, the Island cattle. The objective of this study was to genetically...

  18. Evaluation of cardiac injury biomarkers in cattle with acute clinical mastitis

    Directory of Open Access Journals (Sweden)

    meysam fllah

    2016-05-01

       This study was carried out on 30 Holstein dairy cattle with acute clinical mastitis (ACM and 30 healthy ones. After confirmation of ACM through clinical examination, venous blood samples were collected and cardiac troponin I (cTnI was measured using chemiluminescence assay. Cardiac enzymes activities including CK-MB, AST and LDH were analyzed with special kits and spectrophotometric method. According to the findings mean heart rate (p=0.001, respiratory rate (p=0.026, and rectal temperature (p=0.030 were significantly increased in diseased group. cTnI level was 1.018 ± 0.235 ng/ml in cattle with ACM, which was significantly higher than healthy cattle (0.011±0.006 ng/ml; p=0.000. Other cardiac biomarkers were increased in diseased group, however elevation of serum activities of AST (p=0.047 and CK-MB (p=0.000 were statically significant. Although serum LDH activity in diseased group was higher than control group; but this difference was statistically non-significant (p=0.454. There were significant positive correlations between cTnI concentration with heart rate (p=0.018; r=0.853, respiratory rate (p=0.024; r=0.671, and rectal temperature (p=0.038; r=0.542. Heart rates were significantly correlated with serum activities of CK-MB (p=0.047; r=0.722 and AST (p=0.035; r=0.649. These results indicate some degree of heart damage caused by acute clinical mastitis in dairy cattle.

  19. Determination of selected perfluorinated alkyl acids and persistent organic pollutants from a small volume human serum sample relevant for epidemiological studies.

    Science.gov (United States)

    Koponen, Jani; Rantakokko, Panu; Airaksinen, Riikka; Kiviranta, Hannu

    2013-09-27

    A simple and fast method is presented for the determination of selected persistent organic pollutants (POPs) and perfluorinated alkyl acids (PFAAs), a subgroup of per- and polyfluorinated alkyl substances (PFAS) from a single 200μl aliquot of serum. Sample pretreatment starts with dispersive solid phase extraction of POPs to dichloromethane-hexane, which is immediately poured to cleanup column. POPs are eluted from column and concentrated for GC-MS/MS analysis. PFAAs are trapped to dispersant and are then extracted with ammonium acetate in methanol, concentrated and analysed with LC-MS/MS. For POPs, the limit of detection (LOD) ranged from 1.6 to 17pg/ml. Oxychlordane and dichlorodiphenyltrichloroethane had LODs in the upper end of this range as they were more labile and prone for interferences in the GC-MS/MS. For PFAAs, the LOD range from 0.027 to 0.068ng/ml. For POPs, the accuracy from Standard Reference Materials SRM 1589a and Arctic Monitoring and Assessment Programme (AMAP) intercalibration samples range from 74 to 127% and the repeatability (relative standard deviation, RSD%) from 2.0 to 15%. For PFAAs, the accuracy from AMAP samples ranged from 90 to 110% and from LOQ level spiked serum samples from 72 to 133%. Repeatability from AMAP and LOQ samples ranged from 1.6 to 7.3% and 5.5 to 15%, respectively. The presented method is useful in epidemiological studies where only limited amount of serum is available. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Response of cattle persistently infected with bovine virus diarrhoea virus to bovine leukosis virus.

    Science.gov (United States)

    Roberts, D H; Lucas, M H; Wibberley, G; Westcott, D

    1988-03-26

    Six cattle persistently infected with bovine virus diarrhoea virus (BVDV) and seronegative, and two control, virus negative seropositive cattle were inoculated with lymphocytes infected with bovine leukosis virus (BLV). The two controls produced a normal immune response to BLV, developing antibodies at four and five weeks after inoculation. Two of the six cattle persistently infected with BVDV developed a strong antibody response by six weeks after inoculation with BLV. Four developed a depressed response to BLV, characterised in three by a 'hooking' reaction in the immunodiffusion test which persisted in successive bleedings but was interspersed occasionally by a weak positive reaction. In one of these animals, a series of 'hooking' reactions was followed by a number of negative results. The fourth animal remained serologically negative until 16 weeks after inoculation when a 'hooking' reaction was observed followed by a series of negative results. BLV was isolated from all the cattle persistently infected with BVDV at 42 or 58 weeks after inoculation regardless of whether the serum samples gave negative, 'hooking', weak positive or positive reactions in the immunodiffusion test. BLV was consistently isolated from the nasal secretions of a steer which was BVDV negative but seropositive. The possibility of decreased immune responsiveness to BLV in animals persistently infected with BVDV should be considered when formulating regulations governing the testing of animals for freedom from BLV.

  1. Evaluation of biochemical and ultrasonographic measurements as indicators of undernutrition in cattle

    Directory of Open Access Journals (Sweden)

    S. Strydom

    2008-09-01

    Full Text Available Body condition scoring (BCS gives an indication of the nutritional status of an animal and it is thus an invaluable management tool in domestic livestock systems. It is, however, subjective. This study aimed at identifying biochemical indicators which could be objectively used as an indicator of undernutrition in ruminants. Blood samples were collected from 50 cattle with BCS ≤ 1 and from 50 cattle with BCS ≥ 2.5, using a 0-5 scale, and analysed for albumin, urea, creatinine, fructosamine, beta-hydroxybutyrate, non-esterified fatty acids, total serum protein and haematocrit. Rumpfat and ribfat thickness and marbling relative index were determined ultrasonographically in 15 of the low BCS group and 13 of the high BCS group. The laboratory measure with the best predictive ability for severe undernutrition was albumin, which correctly classified 94 % of cattle, using a cut-off of 31.5 g / ℓ. In contrast to a previous study, our study did not find the fructosamine : albumin ratio to be an accurate test to indicate undernutrition in cattle. Ultrasonic measurement of subcutaneous rumpfat and ribfat proved to reliably predict undernutrition, but may, however be impractical for routine use under most field conditions.

  2. A comparative evaluation of avidin-biotin ELISA and micro SNT for detection of antibodies to infectious bovine rhinotracheitis in cattle population of Odisha, India

    Directory of Open Access Journals (Sweden)

    Priyaranjan Das

    2014-08-01

    Full Text Available Aim: The present study was undertaken to serologically detect Infectious Bovine Rhinotracheitis (IBR in the cattle population of Odisha, India using micro-Serum neutralization test (micro SNT and Avidin-Biotin Enzyme linked immuno sorbent assay (AB ELISA and finding out their comparative efficacy to serve as a suitable diagnostic tool in field condition. Materials and Methods: The study was carried out using serum samples (n=180 collected randomly from cattle populations of nine districts of Odisha. Similarly vaginal swabs (n=26 from cattle having history of repeat breeding, abortion, vulvo-vaginitis and nasal swabs (n=8 from calves with respiratory symptoms and nasal discharge were collected aseptically, to ascertain the circulation of virus among the cattle population. Results: Virus isolation by cell culture and subsequent confirmation by polymerase chain reaction confirmed four isolates. Screening of serum samples revealed 9.44% and 12.22% samples positive for IBR antibodies in micro SNT and AB ELISA respectively. The sensitivity and specificity of AB ELISA test was found to be 88.23% and 95.70% respectively taking micro SNT as gold standard and the kappa value between the two tests was 0.75. Conclusion: Screening of serum samples revealed 9.44% and 12.22% samples positive for IBR antibodies in micro SNT and AB ELISA respectively, thus highlighting the circulation of virus among the livestock population of Odisha and that AB ELISA could be more efficiently applied for the sero-diagnosis of IBR virus infections at field conditions, with demand for more study on faster, efficient and large scale screening of the infected animals.

  3. Determination of carbohydrate-deficient transferrin in human serum with capillary zone electrophoresis. Sample preparation strategies for the removal of interferences caused by increased levels of immunoglobulins.

    Science.gov (United States)

    Lanz, Christian; Falmagne, Jean-Bernard; de l'Escaille, François; Marti, Ulrich; Thormann, Wolfgang

    2008-10-03

    Capillary zone electrophoresis (CZE) in fused-silica capillaries is an effective analytical approach for the separation and determination of the transferrin (Tf) isoforms and thus carbohydrate-deficient transferrin (CDT) in human serum. Sera of patients with progressed liver cirrhosis are prone to interferences in the beta region which prevent the proper determination of CDT by CZE without additional sample preparation. Efforts to identify, reduce or even eliminate these interferences have been undertaken. Data obtained by ultrafiltration, affinity subtraction procedures using protein A, protein L and antibodies against immunoglobulins or Tf, and immunopurification of Tf suggest that the interferences in the patient sera are caused by increased levels of IgA and IgM and are best eliminated by immunopurification. Avian IgY antibody spin column immunocapture of serum Tf followed by CZE analysis of the stripped and concentrated fraction is shown to provide an attractive approach for CDT monitoring in sera with beta region interferences.

  4. Evaluation of trace element status of organic dairy cattle.

    Science.gov (United States)

    Orjales, I; Herrero-Latorre, C; Miranda, M; Rey-Crespo, F; Rodríguez-Bermúdez, R; López-Alonso, M

    2017-11-06

    The present study aimed to evaluate trace mineral status of organic dairy herds in northern Spain and the sources of minerals in different types of feed. Blood samples from organic and conventional dairy cattle and feed samples from the respective farms were analysed by inductively coupled plasma mass spectrometry to determine the concentrations of the essential trace elements (cobalt (Co), chromium (Cr), copper (Cu), iron (Fe), iodine (I), manganese (Mn), molybdenum (Mo), nickel (Ni), selenium (Se) and zinc (Zn)) and toxic trace elements (arsenic (As), cadmium (Cd), mercury (Hg) and lead (Pb)). Overall, no differences between organic and conventional farms were detected in serum concentrations of essential and toxic trace elements (except for higher concentrations of Cd on the organic farms), although a high level of inter-farm variation was detected in the organic systems, indicating that organic production greatly depends on the specific local conditions. The dietary concentrations of the essential trace elements I, Cu, Se and Zn were significantly higher in the conventional than in the organic systems, which can be attributed to the high concentration of these minerals in the concentrate feed. No differences in the concentrations of trace minerals were found in the other types of feed. Multivariate chemometric analysis was conducted to determine the contribution of different feed sources to the trace element status of the cattle. Concentrate samples were mainly associated with Co, Cu, I, Se and Zn (i.e. with the elements supplemented in this type of feed). However, pasture and grass silage were associated with soil-derived elements (As, Cr, Fe and Pb) which cattle may thus ingest during grazing.

  5. The Dutch Brucella abortus monitoring programme for cattle: the impact of false-positive serological reactions and comparison of serological tests.

    Science.gov (United States)

    Emmerzaal, A; de Wit, J J; Dijkstra, Th; Bakker, D; van Zijderveld, F G

    2002-02-01

    The Dutch national Brucella abortus eradication programme for cattle started in 1959. Sporadic cases occurred yearly until 1995; the last infected herd was culled in 1996. In August 1999 the Netherlands was declared officially free of bovine brucellosis by the European Union. Before 1999, the programme to monitor the official Brucella-free status of bovine herds was primarily based on periodical testing of dairy herds with the milk ring test (MRT) and serological testing of all animals older than 1 year of age from non-dairy herds, using the micro-agglutination test (MAT) as screening test. In addition, serum samples of cattle that aborted were tested with the MAT. The high number of false positive reactions in both tests and the serum agglutination test (SAT) and complement fixation test (CFT) used for confirmation seemed to result in unnecessary blockade of herds, subsequent testing and slaughter of animals. For this reason, a validation study was performed in which three indirect enzyme-linked immunosorbent assays (ELISAs), the CFT and the SAT were compared using a panel of sera from brucellosis-free cattle, sera from experimentally infected cattle, and sera from cattle experimentally infected with bacteria which are known to induce cross-reactive antibodies (Pasteurella, Salmonella, Yersinia, and Escherichia). Moreover, four ELISAs and the MRT were compared using a panel of 1000 bulk milk samples from Brucella-free herds and 12 milk samples from Brucella abortus- infected cattle. It is concluded that the ELISA obtained from ID-Lelystad is the most suitable test to monitor the brucelosis free status of herds because it gives rise to fewer false-positive reactions than the SAT.

  6. Anti-Taenia solium metacestode IgG antibodies in serum samples from inhabitants of a central-western region of Brazil

    Directory of Open Access Journals (Sweden)

    Oliveira Heliana B. de

    2006-01-01

    Full Text Available A total of 354 serum samples from inhabitants who frequent the Clinical Laboratory in Catalão, Goiás, in the central-western region of Brazil, were collected from June to August, 2002. The samples were evaluated by indirect immunofluorescence antibody tests and an enzyme linked immunosorbent assay in order to detect anti-Taenia solium metacestode IgG antibodies. Reactive and inconclusive samples were tested by Western blotting (WB. Considering WB as a confirmation, the frequency of antibodies in the serum samples of the above population was 11.3% (CI 5.09 - 17.51. The immunodominant bands most frequently recognized in WB were 64-68 kDa (97.5% and 47-52 kDa (80%. The percentage of seropositivity to cysticercosis was significantly higher for individuals residing in areas without sewage systems (p < 0.0001. In conclusion, the results indicate a probable endemic situation of cysticercosis in this population. These results reinforce the urgent need for control and prevention measures to be taken by the local public health services.

  7. A new method for the determination of benzophenone-UV filters in human serum samples by dispersive liquid-liquid microextraction with liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Vela-Soria, F; Ballesteros, O; Zafra-Gómez, A; Ballesteros, L; Navalón, A

    2014-04-01

    Benzophenone-UV filters (BP-UV filters) are extensively used in cosmetics products to avoid damaging effects of UV radiation. Despite their low toxicity, many research papers indicate that BP-UV filters are weak endocrine disruptors (EDCs). There are clear relationships between BP-UV filters exposure and several health disorders such as carcinogenesis and malformations observed in animals. In the present work, a new sample treatment procedure by dispersive liquid-liquid microextraction (DLLME) is proposed for the extraction of six BPs, namely benzophenone-1 (BP-1), benzophenone-2 (BP-2), benzophenone-3 (BP-3), benzophenone-6 (BP-6), benzophenone-8 (BP-8) and 4-hydroxybenzophenone (4-OH-BP), in human serum samples, followed by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis. The method involves an enzymatic treatment to quantify the total content (free plus conjugated species) of BP-UV filters in serum. The extraction parameters were accurately optimized using multivariate optimization approach. Benzophenone-d₁₀ (BP-d₁₀) was used as surrogate. Limits of quantification (LOQs) ranged from 0.4 to 0.9 ng mL(-1) and inter-day precision (evaluated as relative standard deviation) ranged from 1.9% to 13.1%. The method was validated using matrix-matched calibration and a recovery assay. Recovery rates for spiked samples ranged from 97% to 106%, and acceptable linearity was obtained up to concentrations of 40 ng mL(-1). The method was applied to the determination of the target compounds in human serum samples from 20 randomly selected anonymous individuals. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Coxiella burnetii (Q fever) infection in dairy cattle and associated risk factors in Latvia.

    Science.gov (United States)

    Boroduske, A; Trofimova, J; Kibilds, J; Papule, U; Sergejeva, M; Rodze, I; Grantina-Ievina, L

    2017-07-01

    The purpose of this study was to describe prevalence and spatial distribution of Coxiella burnetii infections in dairy cow sheds in Latvia and to investigate risk factors contributing to C. burnetii infections. Blood serum samples from abortion cases from 1010 sheds have been tested by ELISA for the presence of C. burnetii antibodies and bulk tank milk (BTM) samples from 252 sheds have been tested by real time polymerase chain reaction and ELISA for the presence of C. burnetii DNA and antibodies. Prevalence of C. burnetii antibody-positive sheds in cases of abortion was 13·4%. A total of 10·7% and 13·2% of dairy cow sheds tested positive for the presence of C. burnetii DNA and antibodies in BTM, respectively. Two distinct areas of clustering of test-positive dairy cattle sheds were identified by spatial scan statistics of abortion cases and randomly sampled BTM samples. Three factors were identified as significantly contributing to the risk of C. burnetii DNA presence in BTM - number of cattle in shed (>200 animals/shed) (OR 3·93), location of the shed within risk area in Northern Latvia (OR 8·29) and for the first time, purchasing cattle from abroad has been shown to significantly increase risk (OR 2·68) of C. burnetii infection in dairy cows in Latvia.

  9. Seroprevalence of bovine viral diarrhea virus in crossbred dairy cattle in Bangladesh

    Science.gov (United States)

    Uddin, Mohammed Arif; Ahasan, A. S. M. Lutful; Islam, Kamrul; Islam, Md. Zohorul; Mahmood, Altaf; Islam, Ariful; Islam, Kazi Muhammad Fakhrul; Ahad, Abdul

    2017-01-01

    Aim: The study was conducted to determine the seroprevalence of bovine viral diarrhea virus (BVDV) and hematological features in crossbred dairy cattle in Chittagong, Bangladesh. Materials and Methods: The antibody against BVDV in crossbred dairy cattle serum was detected by indirect enzyme-linked immunosorbent assay. The association of different categorical variables in the prevalence of BVDV has been studied. Blood samples were collected and analyzed to know the hematological variations in the study population. Results: The overall seroprevalence of BVDV in the study area was 51.1% (95% confidence interval [CI], 40.5-61.5). Among different physiological stages of animals, the highest 57.1% (95% CI, 42.2-71.2) prevalence was in case of non-pregnant animals. Aborted cows were found to be significantly (pproduction industry. Therefore, it is necessary to conduct effective control measures to reduce the burden of BVDV. PMID:28919681

  10. Determination of nickel in blood and serum samples of oropharyngeal cancer patients consumed smokeless tobacco products by cloud point extraction coupled with flame atomic absorption spectrometry.

    Science.gov (United States)

    Arain, Sadaf Sadia; Kazi, Tasneem Gul; Arain, Jamshed Bashir; Afridi, Hassan Imran; Kazi, Atif Gul; Nasreen, Syeda; Brahman, Kapil Dev

    2014-10-01

    Oropharyngeal cancer is a significant public health issue in the world. The incidence of oropharyngeal cancer has been increased among people who have habit of chewing smokeless tobacco (SLT) in Pakistan. The aim of present study was to evaluate the concentration of nickel (Ni) in biological samples (whole blood, serum) of oral (n = 95) and pharyngeal (n = 84) male cancer patients. For comparison purposes, the biological samples of healthy age-matched referents (n = 150), who consumed and did not consumed SLT products, were also analyzed for Ni levels. As the Ni level is very low in biological samples, a preconcentration procedure has been developed, prior to analysis of analyte by flame atomic absorption spectrometry (FAAS). The Ni in acid-digested biological samples was complexed with ammonium pyrrolidinedithio carbamate (APDC), and a resulted complex was extracted in a surfactant Triton X-114. Acidic ethanol was added to the surfactant-rich phase prior to its analysis by FAAS. The chemical variables, such as pH, amounts of reagents (APDC, Triton X-114), temperature, incubation time, and sample volume were optimized. The resulted data indicated that concentration of Ni was higher in blood and serum samples of cancer patients as compared to that of referents who have or have not consumed different SLT products (p = 0.012-0.001). It was also observed that healthy referents who consumed SLT products have two to threefold higher levels of Ni in both biological samples as compared to those who were not chewing SLT products (p < 0.01).

  11. Renal amyloidosis in cattle with inflammatory diseases.

    Science.gov (United States)

    Elitok, O M; Elitok, B; Unver, O

    2008-01-01

    The association of inflammatory diseases such as traumatic reticuloperitonitis (TRP), mastitis, metritis, and pododermatitis with renal amyloidosis in cattle is poorly described. Serum amyloid A (SAA) levels are elevated during inflammatory diseases, and renal amyloidosis is formed as a complication. This study was conducted with 82 crossbred cattle with mastitis (n = 18 cows), metritis (n = 11 cows), TRP (n = 30 cows), and pododermatitis (n = 23 : 15 cows and 8 beef cattle). Ten clinically healthy cows served as controls. Hematological, urinary, and blood parameters, including SAA, were measured by an automated procedure provided with trade kits. Determination of amyloidal structures was made by histopathological examination of renal biopsy specimens. At the end of this trial, amyloidosis was detected in 5 cows displaying typical nephrotic syndrome, with hypoproteinemia and proteinuria in combination with polyuria and weight loss. Furthermore, it was observed that cows with renal amyloidosis had significantly higher (P pododermatitis have a high prevalence of systemic amyloidosis in response to inflammation.

  12. Sampling

    CERN Document Server

    Thompson, Steven K

    2012-01-01

    Praise for the Second Edition "This book has never had a competitor. It is the only book that takes a broad approach to sampling . . . any good personal statistics library should include a copy of this book." —Technometrics "Well-written . . . an excellent book on an important subject. Highly recommended." —Choice "An ideal reference for scientific researchers and other professionals who use sampling." —Zentralblatt Math Features new developments in the field combined with all aspects of obtaining, interpreting, and using sample data Sampling provides an up-to-date treat

  13. Influence of lokpa cattle market wastes on agricultural soil quality ...

    African Journals Online (AJOL)

    This paper examined the influence of Lokpa cattle market waste on soil properties. Soil samples were collected from the Central, 3 and 6 m Northwards, Southwards, Eastwards and Westwards of Lokpa cattle market, Umuneochi Local Government Area of Abia State, Nigeria at a depth of 0 to 25 cm (Sample A) and 26 to 50 ...

  14. Molecular prevalence of trypanosome infections in cattle and tsetse flies in the Maasai Steppe, northern Tanzania.

    Science.gov (United States)

    Simwango, Mary; Ngonyoka, Anibariki; Nnko, Happiness J; Salekwa, Linda P; Ole-Neselle, Moses; Kimera, Sharadhuli I; Gwakisa, Paul S

    2017-10-23

    African trypanosomosis is a disease of public health and economic importance that poses a major threat to the livelihoods of people living in the Maasai Steppe, where there is a significant interaction between people, livestock and wildlife. The vulnerability of the Maasai people to the disease is enhanced by the interaction of their cattle, which act as vehicles for trypanosomes, and tsetse flies close to wildlife in protected areas. This study was aimed at identification of trypanosome infections circulating in cattle and tsetse flies in order to understand their distribution and prevalence in livestock/wildlife interface areas in the Maasai Steppe. A total of 1002 cattle and 886 tsetse flies were sampled from June 2015 to February 2016 in five villages and PCR was conducted to amplify the internal transcribed spacer 1 (ITS1) from trypanosomes. All Trypanosoma brucei-positive samples were further tested for the presence of the serum resistance-associated (SRA) gene found in human-infective trypanosomes using the SRA-LAMP technique. The overall prevalence of trypanosome infections was 17.2% in cattle and 3.4% in tsetse flies. Using a nested PCR, prevalence and abundance of five trypanosome species, Trypanosoma vivax, T. brucei, T. simiae, T. theileri and T. congolense, were determined, which varied with season and location. The highest prevalence of the identified trypanosome species was recorded at the end of wet season with an exception of T. brucei which was high at the beginning of the wet season. No human-infective trypanosomes were detected in both cattle and tsetse fly DNA. This study confirms that seasonality and location have a significant contribution to the prevalence of trypanosome species in both mammalian and vector hosts. These results are important for designing of community-wide vector and disease control interventions and planning of sustainable regimes for reduction of the burden of trypanosomosis in endemic pastoral areas, such as the Maasai

  15. Molecular prevalence of trypanosome infections in cattle and tsetse flies in the Maasai Steppe, northern Tanzania

    Directory of Open Access Journals (Sweden)

    Mary Simwango

    2017-10-01

    Full Text Available Abstract Background African trypanosomosis is a disease of public health and economic importance that poses a major threat to the livelihoods of people living in the Maasai Steppe, where there is a significant interaction between people, livestock and wildlife. The vulnerability of the Maasai people to the disease is enhanced by the interaction of their cattle, which act as vehicles for trypanosomes, and tsetse flies close to wildlife in protected areas. This study was aimed at identification of trypanosome infections circulating in cattle and tsetse flies in order to understand their distribution and prevalence in livestock/wildlife interface areas in the Maasai Steppe. Methods A total of 1002 cattle and 886 tsetse flies were sampled from June 2015 to February 2016 in five villages and PCR was conducted to amplify the internal transcribed spacer 1 (ITS1 from trypanosomes. All Trypanosoma brucei-positive samples were further tested for the presence of the serum resistance-associated (SRA gene found in human-infective trypanosomes using the SRA-LAMP technique. Results The overall prevalence of trypanosome infections was 17.2% in cattle and 3.4% in tsetse flies. Using a nested PCR, prevalence and abundance of five trypanosome species, Trypanosoma vivax, T. brucei, T. simiae, T. theileri and T. congolense, were determined, which varied with season and location. The highest prevalence of the identified trypanosome species was recorded at the end of wet season with an exception of T. brucei which was high at the beginning of the wet season. No human-infective trypanosomes were detected in both cattle and tsetse fly DNA. Conclusions This study confirms that seasonality and location have a significant contribution to the prevalence of trypanosome species in both mammalian and vector hosts. These results are important for designing of community-wide vector and disease control interventions and planning of sustainable regimes for reduction of the burden

  16. Identification of extracellular miRNA in archived serum samples by next-generation sequencing from RNA extracted using multiple methods.

    Science.gov (United States)

    Gautam, Aarti; Kumar, Raina; Dimitrov, George; Hoke, Allison; Hammamieh, Rasha; Jett, Marti

    2016-10-01

    miRNAs act as important regulators of gene expression by promoting mRNA degradation or by attenuating protein translation. Since miRNAs are stably expressed in bodily fluids, there is growing interest in profiling these miRNAs, as it is minimally invasive and cost-effective as a diagnostic matrix. A technical hurdle in studying miRNA dynamics is the ability to reliably extract miRNA as small sample volumes and low RNA abundance create challenges for extraction and downstream applications. The purpose of this study was to develop a pipeline for the recovery of miRNA using small volumes of archived serum samples. The RNA was extracted employing several widely utilized RNA isolation kits/methods with and without addition of a carrier. The small RNA library preparation was carried out using Illumina TruSeq small RNA kit and sequencing was carried out using Illumina platform. A fraction of five microliters of total RNA was used for library preparation as quantification is below the detection limit. We were able to profile miRNA levels in serum from all the methods tested. We found out that addition of nucleic acid based carrier molecules had higher numbers of processed reads but it did not enhance the mapping of any miRBase annotated sequences. However, some of the extraction procedures offer certain advantages: RNA extracted by TRIzol seemed to align to the miRBase best; extractions using TRIzol with carrier yielded higher miRNA-to-small RNA ratios. Nuclease free glycogen can be carrier of choice for miRNA sequencing. Our findings illustrate that miRNA extraction and quantification is influenced by the choice of methodologies. Addition of nucleic acid- based carrier molecules during extraction procedure is not a good choice when assaying miRNA using sequencing. The careful selection of an extraction method permits the archived serum samples to become valuable resources for high-throughput applications.

  17. Development of a novel monolith frit-based solid-phase microextraction method for determination of hexanal and heptanal in human serum samples.

    Science.gov (United States)

    Xu, Hui; Yan, Zhihua; Song, Dandan

    2012-03-01

    In this paper, a polypropylene frit with porous network structure and high area-to-thickness ratio (4.8 mm diameter, 1.6 mm thickness, 20 mm pore size) was utilized as a mould of monolith. Poly(methacrylic acid-ethlyene glycol dimethacrylate) (MAA-EGDMA) monolith was in situ synthesized in the micro-channel of frit by photopolymerization. A monolith frit-based solid-phase microextraction method (SPME) was developed for the determination of hexanal and heptanal in serum samples by combining with high-performance liquid chromatography. 2,4-Dinitrophenylhydrazine (DNPH) as the derivatizing reagent was absorbed on a monolith frit, then its derivatization reaction with aldehydes and the absorption of formed hydrazones on the monolith disk occurred simultaneously. The condition parameters for polymerization, derivatization and extraction were optimized systematically. Under the optimum conditions, rigid structure, low back-pressure and high column capacity were achieved for the monolith frit. The limits of detection for hexanal and heptanal were 1.86 and 1.38 nmol/L, respectively. The inter- and intra-day relative standard deviations were less than 7.7% (n = 6). This method was applied successfully to aldehydes analysis in human serum samples. The method possesses advantages such as simplicity, efficiency, low cost and good biocompatibility. It provides an alternative approach for quantification of aldehydes in complex biological samples. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Naturally occurring infections of cattle with Theileria lestoquardi and sheep with Theileria annulata in the Sudan

    National Research Council Canada - National Science Library

    Taha, K M; Salih, D A; Ali, A M; Omer, R A; El Hussein, A M

    2013-01-01

    .... In an attempt to evaluate field cross infectivity of Theileria lestoquardi and T. annulata in cattle and sheep respectively, a PCR analysis was carried out on samples collected from closely reared sheep and cattle using both T. annulata...

  19. Shedding and seroprevalence of pathogenic Leptospira spp. in sheep and cattle at a New Zealand Abattoir.

    Science.gov (United States)

    Fang, F; Collins-Emerson, J M; Cullum, A; Heuer, C; Wilson, P R; Benschop, J

    2015-06-01

    A cross-sectional study was carried out on sheep and cattle slaughtered at a New Zealand abattoir from September to November 2010 to investigate the supplier-specific shedding rate, renal carriage rate and seroprevalence of leptospires. In the 2008/2009 season, this abattoir experienced three human leptospirosis cases from 20 staff, of which two were hospitalized. Urine, kidney and blood samples were collected from carcasses of 399 sheep (six suppliers, 17 slaughter lines) and 146 cattle (three suppliers, 22 slaughter lines). The urine and kidney samples were tested by quantitative real-time PCR (qPCR), while serum samples (from coagulated blood samples) were tested by microscopic agglutination test (MAT). In total, 27% (73/274; 95% CI: 18-37) of urine samples tested positive by qPCR. Species-specific shedding rates (prevalence of positive urine qPCR) were 31% (95% CI: 17-48) for sheep and 21% (95% CI: 14-30) for cattle. For 545 kidney samples tested, 145 were qPCR positive (27%; 95% CI: 17-39). The average prevalence of kidney qPCR positivity was 29% (95% CI: 17-45) for sheep and 21% (95% CI: 15-28) for cattle. Three hundred and thirty of 542 sampled sheep and cattle had antibodies against Leptospira borgpetersenii serovar Hardjobovis (Hardjobovis) and/or Leptospira interrogans serovar Pomona (Pomona), based on reciprocal MAT titre ≥1 : 48 (overall seroprevalence of 61%; 95% CI: 48-73). Seroprevalence was 57% (95% CI: 40-72) for sheep and 73% (95% CI: 59-83) for cattle. Among the seropositive animals, 41% (70/170; 95% CI: 30-54) were shedding (tested positive by urine qPCR) and 42% (137/330; 95% CI: 30-54) had renal carriage (tested positive by kidney qPCR). Some risk management options for abattoirs or farms to prevent human leptospirosis infections include vaccination of maintenance hosts, the use of personal protective equipment, and the application of urine qPCR to detect shedding status of stock as surveillance and as an alert. © 2014 Blackwell Verlag

  20. Local and global patterns of admixture and population structure in Iranian native cattle.

    Science.gov (United States)

    Karimi, Karim; Strucken, Eva M; Moghaddar, Nasir; Ferdosi, Mohammad H; Esmailizadeh, Ali; Gondro, Cedric

    2016-07-15

    Two separate domestication events gave rise to humped zebu cattle in India and humpless taurine cattle in the Fertile Crescent of the Near and Middle East. Iran covers the Eastern side of the Fertile Crescent and exhibits a variety of native cattle breeds, however, only little is known about the admixture patterns of Iranian cattle and their contribution to the formation of modern cattle breeds. Genome-wide data (700 k chip) of eight Iranian cattle breeds (Sarabi N = 19, Kurdi N = 7, Taleshi N = 7, Mazandarani N = 10, Najdi N = 7, Pars N = 7, Kermani N = 9, and Sistani N = 9) were collected from across Iran. For a local assessment, taurine (Holstein and Jersey) and indicine (Brahman) outgroup samples were used. For the global perspective, 134 world-wide cattle breeds were included. Between breed variation amongst Iranian cattle explained 60 % (p populations most accurately explained the admixture of 44 selected representative cattle breeds (standard error 0.4617). Low levels of African ancestry were identified in Iranian cattle breeds (on average 7.5 %); however, the signal did not persist through all analyses. Admixture and migration analyses revealed minimal introgression from Iranian cattle into other taurine cattle (Holstein, Hanwoo, Anatolian breeds). The eight Iranian cattle breeds feature a discrete genetic composition which should be considered in conservation programs aimed at preserving unique species and genetic diversity. Despite a complex admixture pattern among Iranian cattle breeds, there was no strong introgression from other world-wide cattle breeds into Iranian cattle and vice versa. Considering Iran's central location of cattle domestication, Iranian cattle might represent a local domestication event that remained contained and did not contribute to the formation of modern breeds, or genetics of the ancestral population that gave rise to modern cattle is too diluted to be linked directly to any current cattle

  1. Detection of human parvovirus B19 in serum samples from children under 5 years of age with rash-fever illnesses in the Democratic Republic of the Congo.

    Science.gov (United States)

    Wawina, Tony Bokalanga; Tshiani, Olivier Mbaya; Ahuka, Steve Mundeke; Pukuta, Elisabeth Simbu; Aloni, Michel Ntetani; Kasanga, Christopher Jacob; Muyembe, Jean-Jacques Tamfum

    2017-12-01

    It has been demonstrated that infection with human parvovirus B19 (B19V) is associated with rash-fever illnesses. The present study aimed to investigate B19V as an aetiological agent of rash-fever syndromes in Congolese children confirmed as measles and rubella IgM-negative. An ELISA IgM test and PCR were performed to screen for B19V. A total of 177 archived serum samples were randomly selected from the measles biobank of the National Institute for Biomedical Research (INRB). Samples were investigated for anti-B19V IgM and B19V DNA. These samples originated from children <5years of age with measles-like rashes, previously confirmed as negative for both measles and rubella IgM. Out of 177 serum samples tested by ELISA and 168 tested by PCR, 109 were positive for B19V IgM antibodies (61.6%) and 87 (51.8%) were positive for B19V DNA. Positive samples in both assays were from all provinces of DRC. B19V plays a role in rash-fever illnesses in children under 5 years of age suspected of having measles or rubella infections in DRC. As an aetiological cause of rash and fever syndromes, the present study demonstrates that B19V should also be considered during the laboratory investigation of rash-fever illnesses in DRC, particularly in the paediatric population. There is a need to conduct further studies in order to gain a better understanding of the spatiotemporal pattern of B19V and to define the genotype(s) of B19V circulating in DRC. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  2. An optical sensor for the determination of digoxin in serum samples based on a molecularly imprinted polymer membrane

    Energy Technology Data Exchange (ETDEWEB)

    Paniagua Gonzalez, Gema [Departamento de Ciencias Analiticas, Facultad de Ciencias, Universidad Nacional de Educacion a Distancia (UNED), 28040 Madrid (Spain); Fernandez Hernando, Pilar, E-mail: pfhernando@ccia.uned.es [Departamento de Ciencias Analiticas, Facultad de Ciencias, Universidad Nacional de Educacion a Distancia (UNED), 28040 Madrid (Spain); Durand Alegria, J. S. [Departamento de Ciencias Analiticas, Facultad de Ciencias, Universidad Nacional de Educacion a Distancia (UNED), 28040 Madrid (Spain)

    2009-04-13

    This paper reports the synthesis and testing of a molecularly imprinted polymer membrane for digoxin analysis. Digoxin-specific bulk polymer was obtained by the UV initiated co-polymerisation of methacrylic acid and ethylene glycol dimethacrylate in acetonitrile as porogen. After extracting the template analyte, the ground polymer particles were mixed with plasticizer polyvinyl chloride to form a MIP membrane. A reference polymer membrane was prepared from the same mixture of monomers but with no template. The resultant membrane morphologies were examined by scanning electron microscopy. The imprinted membrane was tested as the recognition element in a digoxin-sensitive fluorescence sensor; sensor response was measured using standard solutions of digoxin at concentrations of up to 4 x 10{sup -3} mg L{sup -1}. The detection limit was 3.17 x 10{sup -5} mg L{sup -1}. Within- and between-day relative standard deviations RSD (n = 5) were in the range 4.5-5.5% and 5.5-6.5% respectively for 0 and 1 x 10{sup -3} mg L{sup -1} digoxin concentrations. A selectivity study showed that compounds of similar structure to digoxin did not significantly interfere with detection for interferent concentrations at 10, 30 and 100 times higher than the digoxin concentration. This simply manufactured MIP membrane showed good recognition characteristics, a high affinity for digoxin, and provided satisfactory results in analyses of this analyte in human serum.

  3. A novel strategy for spectrophotometric simultaneous determination of amitriptyline and nortriptyline based on derivation with a quinonoid compound in serum samples

    Science.gov (United States)

    Farnoudian-Habibi, Amir; Massoumi, Bakhshali; Jaymand, Mehdi

    2016-11-01

    A novel and efficient strategy for the simultaneous determination of two tricyclic antidepressant (TCA) drugs [amitriptyline (AT), and its main metabolite (nortriptyline; NT)] via a combination of magnetic solid phase extraction (MSPE), and spectrophotometric techniques in serum is suggested. For this purpose, the imidazolium ionic liquid (Imz)-modified Fe3O4@SiO2 nanoparticles (Fe3O4@SiO2-Imz) was employed as an adsorbent for the MSPE. Preconcentration (loading-desorption) studies were performed under optimized conditions including pH, adsorbent amount, contact time, eluent volume, and desorption time. Afterward, determination of each drug was carried out by specific strategy. Acetaldehyde (AC), and 2,3,5,6-tetrachloro-1,4-benzoquinone (chloranil; CL) were used as chemical reagents for reaction with NT, while AT did not react with these reagents. This method is based on the condensation reaction between secondary amine group of NT and AC to afford an enamine, and subsequently reaction with CL to produce a chlorinated quinone-substituted enamine. The final product exhibited maximum absorption at 556 nm, while the AT was determined at 240 nm. The limits of detections (LODs) for NT and AT in serum sample were obtained as 0.19 and 0.90 ng mL- 1, respectively. The limits of quantifications (LOQs) were obtained to be 0.63 and 2.93 ng mL- 1 for NT and AT, respectively. A linear range was obtained to be 1 to 5 ng mL- 1. Results indicated that the suggested method is applicable for simultaneous determination of NT and AT in serum samples.

  4. Evaluation of a feline-specific multiplex, bead-based assay for detection of cytokines, chemokines, growth factors, and other immunologically active proteins in serum and plasma samples from cats.

    Science.gov (United States)

    Halpin, Rachel E; Saunders, Rebecca S; Thompson, Beverly J; Rohde Newgent, Allison S; Amorim, Juliana; Melillo, Gabrielle N; DeClue, Amy E

    2016-05-01

    OBJECTIVE To evaluate a feline-specific multiplex, bead-based assay system for detection of recombinant and native proteins in serum samples and in EDTA-treated and heparinized plasma samples. SAMPLE Serum samples and EDTA-treated and heparinized plasma samples from 30 sick cats and 9 healthy client-owned cats and heparinized whole blood samples from 5 healthy purpose-bred cats. PROCEDURES Ability of the assay system to detect 19 recombinant and native immunologically active proteins in plasma and serum samples from healthy and purpose-bred cats was evaluated via spike-and-recovery tests, assessments of inter- and intra-assay variation, linearity results, and leukocyte stimulation. Effects of various concentrations of heparin and serum matrix solution on percentages of analytes recovered were also evaluated. Analyte concentrations in samples from healthy and sick cats were measured and compared between groups. RESULTS Percentages of analytes recovered were unsatisfactory for most assays. Serum and heparinized plasma samples yielded better recovery results than did EDTA-treated plasma samples. Use of serum matrix solution did not improve results. Use of heparin concentrations greater than the recommended range affected the results. Linearity of results was difficult to assess because of the poor recovery. For the analytes that were recovered sufficiently for assessment, linearity appeared to be reasonable despite the limited detection. CONCLUSIONS AND CLINICAL RELEVANCE Poor percentages of analytes recovered and adverse effects of sample protein matrix limited the usefulness of the multiplex, bead-based assay system for measurement of immunologically active proteins in solutions with high protein content; however, recovery results were fairly linear, potentially allowing evaluation of feline plasma or serum samples with high analyte concentrations.

  5. Development of a candidate reference measurement procedure for the analysis of cortisol in human serum samples by isotope dilution-gas chromatography-mass spectrometry.

    Science.gov (United States)

    Kawaguchi, Migaku; Takatsu, Akiko

    2009-08-01

    A candidate reference measurement procedure involving isotope dilution coupled with gas chromatography-mass spectrometry (GC-MS) has been developed and critically evaluated. An isotopically labeled internal standard, cortisol-d(2), was added to a serum sample. After equilibration, solid-phase extractions (SPE) for sample preparation and derivatization with heptafluorobutyric anhydride (HFBA) were performed for GC-MS analysis. The limit of detection (LOD) and the limit of quantification (LOQ) were 5 and 20 ng g(-1), respectively. The recovery of the added cortisol ranged from 99.8 to 101.0%. Excellent precision was obtained with a within-day variation (RSD) of 0.7% for GC-MS analysis. The accuracy of the measurement was evaluated by comparing of results of this reference measurement procedure on lyophilized human serum reference materials for cortisol (European Reference Materials (ERM)-DA 192) as Certified Reference Materials (CRMs). The results of this method for total cortisol agreed with the certified values within some uncertainty. This method, which demonstrates simply, easy, good accuracy, high precision, and is free from interferences from structural analogues, qualifies as a reference measurement procedure.

  6. The detergent fraction is effective in the detection of IgG anti-Strongyloides stercoralis in serum samples from immunocompromised individuals.

    Science.gov (United States)

    da Silva, Herculano; de Carvalho, Célio José Victal; Levenhagen, Marcelo Arantes; Costa-Cruz, Julia Maria

    2014-12-01

    Human strongyloidiasis is an intestinal helminthiasis that can be fatal particularly in cases of immunosuppression. The aim of this study is to assess the diagnostic accuracy of the detergent fraction (D), purified from total saline extract (SE) of Strongyloides venezuelensis, in the detection of anti-Strongyloides stercoralis IgG antibodies in serum samples from individuals coming from endemic areas for strongyloidiasis and presenting immunocompromised conditions: human immunodeficiency virus (HIV(+)), diabetes mellitus type 2, cancer, tuberculosis and alcoholism. Serum samples from 93 individuals were analyzed by ELISA, as follows: Group 1: 30 immunocompromised individuals with strongyloidiasis; Group 2: 33 immunocompromised individuals without strongyloidiasis and Group 3: 30 healthy individuals. The total saline extract (SE) and detergent fraction (D) showed a sensitivity of 73.33 and 83.33%, and specificity of 82.15 and 86.36%, respectively. The detergent fraction was effective to detect anti-S. stercoralis IgG antibodies in immunocompromised individuals with strongyloidiasis and may be applied as an important tool in the immunodiagnosis of human strongyloidiasis related to immunosuppression. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  7. Total α1-acid glycoprotein determination in serum samples using disposable screen-printed electrodes and osmium (VI) as electrochemical tag.

    Science.gov (United States)

    Sierra, Tania; González, María Cristina; Moreno, Begoña; Crevillen, Agustin G; Escarpa, Alberto

    2018-04-01

    Alpha-1-acid glycoprotein (AGP) or Orosomucoid is a serum glycoprotein which belongs to the group of acute phase proteins. It is a potential biomarker for inflammatory bowel diseases. In this sense, there is a need for developing fast and cheap analytical methods for diagnosis, prognosis and follow-up of these diseases. In this work, we propose a simple and cheap electrochemical method for total AGP determination using disposable carbon screen-printed electrodes (SPE) and using a selective acidic precipitation of the rest of proteins. This method avoids the use of biological components, decreasing dramatically the analysis cost. Firstly, AGP is labeled with an electrochemical tag (osmium (VI) complex) and then the total amount of AGP is quantified by adsorptive transfer stripping square wave voltammetry. The method optimized showed a good linear correlation (r = 0.9992) and limit of detection of 1.6mgl-1. The methodology was successfully applied to quantify AGP in a commercial serum sample. This methodology could be useful in clinical diagnosis because of AGP levels increase two or three times when inflammatory processes happen. Moreover, the inherent advantages of SPE technology (low sample consumption, low cost and point of care testing) make this methodology very attractive in this field. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Matrix removal in state of the art sample preparation methods for serum by charged aerosol detection and metabolomics-based LC-MS.

    Science.gov (United States)

    Schimek, Denise; Francesconi, Kevin A; Mautner, Anton; Libiseller, Gunnar; Raml, Reingard; Magnes, Christoph

    2016-04-07

    Investigations into sample preparation procedures usually focus on analyte recovery with no information provided about the fate of other components of the sample (matrix). For many analyses, however, and particularly those using liquid chromatography-mass spectrometry (LC-MS), quantitative measurements are greatly influenced by sample matrix. Using the example of the drug amitriptyline and three of its metabolites in serum, we performed a comprehensive investigation of nine commonly used sample clean-up procedures in terms of their suitability for preparing serum samples. We were monitoring the undesired matrix compounds using a combination of charged aerosol detection (CAD), LC-CAD, and a metabolomics-based LC-MS/MS approach. In this way, we compared analyte recovery of protein precipitation-, liquid-liquid-, solid-phase- and hybrid solid-phase extraction methods. Although all methods provided acceptable recoveries, the highest recovery was obtained by protein precipitation with acetonitrile/formic acid (amitriptyline 113%, nortriptyline 92%, 10-hydroxyamitriptyline 89%, and amitriptyline N-oxide 96%). The quantification of matrix removal by LC-CAD showed that the solid phase extraction method (SPE) provided the lowest remaining matrix load (48-123 μg mL(-1)), which is a 10-40 fold better matrix clean-up than the precipitation- or hybrid solid phase extraction methods. The metabolomics profiles of eleven compound classes, comprising 70 matrix compounds showed the trends of compound class removal for each sample preparation strategy. The collective data set of analyte recovery, matrix removal and matrix compound profile was used to assess the effectiveness of each sample preparation method. The best performance in matrix clean-up and practical handling of small sample volumes was showed by the SPE techniques, particularly HLB SPE. CAD proved to be an effective tool for revealing the considerable differences between the sample preparation methods. This detector can

  9. A competitive immunoassay for ultrasensitive detection of Hg{sup 2+} in water, human serum and urine samples using immunochromatographic test based on surface-enhanced Raman scattering

    Energy Technology Data Exchange (ETDEWEB)

    She, Pei; Chu, Yanxin [The Key Lab of Health Chemistry & Molecular Diagnosis of Suzhou, College of Chemistry, Chemical Engineering & Materials Science, Soochow University, Renai Road 199, Suzhou 215123 (China); Liu, Chunwei; Guo, Xun [OptoTrace (Suzhou) Technologies, Inc., STE 316, Building 4, No. 218, Xinghu Street, bioBAY, Suzhou Industrial Park, Suzhou 215123 (China); Zhao, Kang [The Key Lab of Health Chemistry & Molecular Diagnosis of Suzhou, College of Chemistry, Chemical Engineering & Materials Science, Soochow University, Renai Road 199, Suzhou 215123 (China); Li, Jianguo, E-mail: lijgsd@suda.edu.cn [The Key Lab of Health Chemistry & Molecular Diagnosis of Suzhou, College of Chemistry, Chemical Engineering & Materials Science, Soochow University, Renai Road 199, Suzhou 215123 (China); Du, Haijing; Zhang, Xiang [The Key Lab of Health Chemistry & Molecular Diagnosis of Suzhou, College of Chemistry, Chemical Engineering & Materials Science, Soochow University, Renai Road 199, Suzhou 215123 (China); Wang, Hong [OptoTrace (Suzhou) Technologies, Inc., STE 316, Building 4, No. 218, Xinghu Street, bioBAY, Suzhou Industrial Park, Suzhou 215123 (China); Deng, Anping, E-mail: denganping@suda.edu.cn [The Key Lab of Health Chemistry & Molecular Diagnosis of Suzhou, College of Chemistry, Chemical Engineering & Materials Science, Soochow University, Renai Road 199, Suzhou 215123 (China)

    2016-02-04

    An immunochromatographic test (ICT) strip was developed for ultrasensitive competitive immunoassay of Hg{sup 2+}. This strategy was achieved by combining the easy-operation and rapidity of ICT with the high sensitivity of surface-enhanced Raman scattering (SERS). Monoclonal antibody (mAb) against Hg{sup 2+} and Raman active substance 4-mercaptobenzoic acid (MBA) dual labelled gold nanoparticles (GNPs) were prepared as an immunoprobe. The Raman scattering intensity of MBA on the test line of the ICT strip was measured for quantitative determination of Hg{sup 2+}. The ICT was able to directly detect Hg{sup 2+} without complexing due to the specific recognition of the mAb with Hg{sup 2+}. The IC{sub 50} and limit of detection (LOD) of the assay for Hg{sup 2+} detection were 0.12 ng mL{sup −1} and 0.45 pg mL{sup −1}, respectively. There was no cross-reactivity (CR) of the assay with other nineteen ions and the ICT strips could be kept for 5 weeks without loss of activity. The recoveries of the assay for water, human serum and urine samples spiked with Hg{sup 2+} were in range of 88.3–107.3% with the relative standard deviations (RSD) of 1.5–9.5% (n = 3). The proposed ICT was used for the detection of Hg{sup 2+} in urine samples collected from Occupational Disease Hospital and the results were confirmed by cold-vapor atomic fluorescence spectroscopy (CV-AFS). The assay exhibited high sensitivity, selectivity, stability, precision and accuracy, demonstrating a promising method for the detection of trace amount of Hg{sup 2+} in environmental water samples and biological serum and urine samples. - Highlights: • The proposed ICT was able to directly detect Hg{sup 2+} without formation of Hg{sup 2+}-ligand complex. • The proposed ICT exhibited high sensitivity, specificity, stability, precision and accuracy for Hg{sup 2+} detection. • The proposed ICT was applicable for the detection of trace amount of Hg{sup 2+} in water, human serum and urine samples.

  10. Serum iron test

    Science.gov (United States)

    Fe+2; Ferric ion; Fe++; Ferrous ion; Iron - serum; Anemia - serum iron; Hemochromatosis - serum iron ... A blood sample is needed. Iron levels are highest in the morning. Your health care provider will likely have you do this test in the morning.

  11. Sample preparation-free, real-time detection of microRNA in human serum using piezoelectric cantilever biosensors at attomole level.

    Science.gov (United States)

    Johnson, Blake N; Mutharasan, Raj

    2012-12-04

    A sensitive, selective, sample preparation-free method for near real-time detection of microRNA in buffer and human serum is given using gold (Au)-coated dynamic piezoelectric cantilever sensors. Sensor response to thiolated DNA probe chemisorption, hsa-let-7a hybridization, labeled-DNA hybridization, and Au nanoparticle-functionalized DNA hybridization was monitored continuously in flowing liquid samples using custom flow-cells. The assay showed successful detection of target let-7a with a dynamic range spanning 6 orders of magnitude (10 fM-1 nM) with a limit of detection of less than 10 attomoles (∼4 fM). The serum background had negligible effect on sensitivity relative to the results obtained in the buffer due to reduction in nonspecific binding caused by continuous sensor vibration. Both hybridization and nonspecific binding reduction were confirmed using fluorescence-based assays to support sensor-based results. The sensor-based method demonstrated excellent selectivity for the microRNA target in comparison with similar microRNA differing by only a single nucleotide (hsa-let-7c) and random microRNA sequences. Au nanoparticle-based amplification of sensor response was investigated and led to an order of magnitude improvement in the detection limit and a 128% amplification of sensor response over the entire dynamic range. Au nanoparticle amplification was verified by scanning electron microscopy. The cantilever sensor-based microRNA assay provides competitive sensitivity with current microRNA detection methods and has the advantage of requiring no sample preparation, even when working with biological samples that contain a complex background.

  12. Significance of Neospora caninum in cattle farming

    OpenAIRE

    Ilić Tamara; Dimitrijević Sanda

    2006-01-01

    Neospora caninum is an obligate intracellular protozoan parasite which primarily causes diseases in dogs and cattle all over the world. It was first described in Norway in the mid-eighties in dogs, after which, until the present time, clinical neosporosis was proven in sheep, goats, deer, rhinoceroses, horses, and experimental rodents. Antibodies against N. caninum have been found also in the serum of water buffalo, red and gray foxes, coyotes, camels, and felines. Due to the similarity of th...

  13. Development of an enzyme-linked immunosorbent assay for serodiagnosis of ringworm infection in cattle.

    Science.gov (United States)

    Bagut, Elena Tatiana; Cambier, Ludivine; Heinen, Marie-Pierre; Cozma, Vasile; Monod, Michel; Mignon, Bernard

    2013-08-01

    The aim of this study was to develop an in-house enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of ringworm infection in cattle. We used available recombinant forms of Trichophyton rubrum dipeptidyl peptidase V (TruDppV) and T. rubrum leucin aminopeptidase 2 (TruLap2), which are 98% identical to Trichophyton verrucosum orthologues. Field serum samples from 135 cattle with ringworm infection, as confirmed by direct microscopy, fluorescence microscopy, and PCR, and from 55 cattle without any apparent skin lesions or history of ringworm infection that served as negative controls were used. Sensitivities, specificities, and positive and negative predictive values were determined to evaluate the diagnostic value of our ELISA. Overall, the ELISAs based on recombinant TruDppV and TruLap2 discriminated well between infected animals and healthy controls. Highly significant differences (P < 0.0001, Mann-Whitney U test) were noted between optical density values obtained when sera from infected versus control cattle were tested. The ELISA developed for the detection of specific antibodies against DppV gave 89.6% sensitivity, 92.7% specificity, a 96.8% positive predictive value, and a 78.4% negative predictive value. The recombinant TruLap2-based ELISA displayed 88.1% sensitivity, 90.9% specificity, a 95.9% positive predictive value, and a 75.7% negative predictive value. To the best of our knowledge, this is the first ELISA based on recombinant antigens for assessing immune responses to ringworm infection in cattle; it is particularly suitable for epidemiological studies and also for the evaluation of vaccines and/or vaccination procedures.

  14. Collection and Characterization of Samples for Establishment of a Serum Repository for Lyme Disease Diagnostic Test Development and Evaluation

    Science.gov (United States)

    Molins, Claudia R.; Sexton, Christopher; Young, John W.; Ashton, Laura V.; Pappert, Ryan; Beard, Charles B.

    2014-01-01

    Serological assays and a two-tiered test algorithm are recommended for laboratory confirmation of Lyme disease. In the United States, the sensitivity of two-tiered testing using commercially available serology-based assays is dependent on the stage of infection and ranges from 30% in the early localized disease stage to near 100% in late-stage disease. Other variables, including subjectivity in reading Western blots, compliance with two-tiered recommendations, use of different first- and second-tier test combinations, and use of different test samples, all contribute to variation in two-tiered test performance. The availability and use of sample sets from well-characterized Lyme disease patients and controls are needed to better assess the performance of existing tests and for development of improved assays. To address this need, the Centers for Disease Control and Prevention and the National Institutes of Health prospectively collected sera from patients at all stages of Lyme disease, as well as healthy donors and patients with look-alike diseases. Patients and healthy controls were recruited using strict inclusion and exclusion criteria. Samples from all included patients were retrospectively characterized by two-tiered testing. The results from two-tiered testing corroborated the need for novel and improved diagnostics, particularly for laboratory diagnosis of earlier stages of infection. Furthermore, the two-tiered results provide a baseline with samples from well-characterized patients that can be used in comparing the sensitivity and specificity of novel diagnostics. Panels of sera and accompanying clinical and laboratory testing results are now available to Lyme disease serological test users and researchers developing novel tests. PMID:25122862

  15. Vitamin D status of dairy cattle: Outcomes of current practices in the dairy industry.

    Science.gov (United States)

    Nelson, Corwin D; Lippolis, John D; Reinhardt, Timothy A; Sacco, Randy E; Powell, Jessi L; Drewnoski, Mary E; O'Neil, Matthew; Beitz, Donald C; Weiss, William P

    2016-12-01

    The need for vitamin D supplementation of dairy cattle has been known for the better part of the last century and is well appreciated by dairy producers and nutritionists. Whether current recommendations and practices for supplemental vitamin D are meeting the needs of dairy cattle, however, is not well known. The vitamin D status of animals is reliably indicated by the concentration of the 25-hydroxyvitamin D [25(OH)D] metabolite in serum or plasma, with a concentration of 30ng/mL proposed as a lower threshold for sufficiency. The objective of this study was to determine the typical serum 25(OH)D concentrations of dairy cattle across various dairy operations. The serum 25(OH)D concentration of 702 samples collected from cows across various stages of lactation, housing systems, and locations in the United States was 68±22ng/mL (mean ± standard deviation), with the majority of samples between 40 and 100ng/mL. Most of the 12 herds surveyed supplemented cows with 30,000 to 50,000 IU of vitamin D3/d, and average serum 25(OH)D of cows at 100 to 300 DIM in each of those herds was near or above 70ng/mL regardless of season or housing. In contrast, average serum 25(OH)D of a herd supplementing with 20,000 IU/d was 42±15ng/mL, with 22% below 30ng/mL. Cows in early lactation (0 to 30d in milk) also had lower serum 25(OH)D than did mid- to late-lactation cows (57±17 vs. 71±20ng/mL, respectively). Serum 25(OH)D of yearling heifers receiving 11,000 to 12,000 IU of vitamin D3/d was near that of cows at 76±15ng/mL. Serum 25(OH)D concentrations of calves, on the other hand, was 15±11ng/mL at birth and remained near or below 15ng/mL through 1mo of age if they were fed pasteurized waste milk with little to no summer sun exposure. In contrast, serum 25(OH)D of calves fed milk replacer containing 6,600 and 11,000 IU of vitamin D2/kg of dry matter were 59±8 and 98±33ng/mL, respectively, at 1mo of age. Experimental data from calves similarly indicated that serum 25(OH

  16. Heterogeneous results of serum testosterone obtained in multiple blood samples from women with idiopathic hirsutism in both basal and post-dexamethasone conditions.

    Science.gov (United States)

    Scaglia, H E; Aquilano, D; Zylbersztein, C; Spinedi, E; Colombani, M; Simon, J; Encabo, J

    1981-01-01

    Serum radioimmunoassayable testosterone (T), cortisol and luteinizing hormone (LH) were determined before and after dexamethasone (DXM) administration in 35 patients with idiopathic hirsutism (IH). Blood samples were taken at 15-min intervals during 1 hour in both basal and post-DXM conditions. Testosterone values obtained in 5 normal women in the same conditions during the early follicular phase were (mean +/- SD): baseline, 2.26 +/- 0.49 nmol/l; post-DXM, 0.80 +/- 0.35 nmol/l. Serum T levels in the whole group of patients with IH were significantly higher than those in the control group (mean +/- SD): baseline, 3.30 +/- 1.80nmol/l; post-DXM, 1.67 +/- 1.49nmol/l. Patients with IH were divided into 4 groups according to T results in the DXM test (mean +/- SD in both basal and post-DXM conditions, respectively): group 1 (n = 13) 1.67 +/- 0.66 and 0.62 +/- 0.35nmol/l; group 2 (n = 11) 3.89 +/- 1.63 and 3.09 +/- 1.49nmol/l; group 3 (n = 6) 3.96 +/- 1.46 and 0.87 +/- 0.73nmol/l; and group 4 (n = 5) 5.45 +/- 1.25 and 2.05 +/- 0.38nmol/l. In all cases, maximal adrenal inhibition, as judged by serum cortisol, was obtained. No LH modifications after DXM were obtained in any of the cases. Our results demonstrate that there is no common androgenic abnormality in IH. It is possible to obtain normal or high circulating T levels. The findings of this study also suggest that the adrenals, to ovary or both may be the sources of high T levels.

  17. Measurement of yunaconitine and crassicauline A in small-volume blood serum samples by LC-MS/MS: tracing of aconite poisoning in clinical diagnosis.

    Science.gov (United States)

    Ka-Wing Chung, Karen; Pak-Lam Chen, Sammy; Ng, Sau-Wah; Wing-Lai Mak, Tony; Sze-Yin Leung, Kelvin

    2012-08-15

    Aconite poisoning is one of the most serious types of herb-related medical emergencies. In Hong Kong, many if not most of these poisoning cases are due to confusion in herbal species; that is, the wrong herbs are used in prescriptions. Such human errors, while inevitable perhaps, can be serious, and sometimes fatal. The chemical components responsible for aconite poisoning are yunaconitine and crassicauline A. In the present study, a rapid and sensitive method for the screening and quantification of yunaconitine and crassicauline A in human serum, using LC-MS/MS, was developed and validated. Methyllycaconitine was chosen as the internal standard. The limit of detection (LOD) of yunaconitine and crassicauline A were found to be 0.022 and 0.021 ng/mL, respectively. The limit of quantification (LOQ) was 0.1 ng/mL for both yunaconitine and crassicauline A. The recovery of yunaconitine and crassicauline A ranged from 78.6% to 84.9% and 78.3% to 87.2%, respectively. The matrix effect of yunaconitine and crassicauline A ranged from 110.0% to 130.4% and 121.2 to 130.0%, respectively. Both yunaconitine and crassicauline A were stable in serum for at least 3 months at -20 °C, and the extracts were stable for at least 7 days. For clinical applications, serum samples of two patients confirmed to have had aconite herbs poisoning in 2008 were quantified using the developed method. The result showed that this method can be utilized in clinical routine applications. This screening method expedites the diagnosis in cases of suspected aconite poisoning, thus enabling doctors to treat the condition more quickly and effectively. Copyright © 2012 Elsevier B.V. All rights reserved.

  18. Ochratoxin A in human serum samples collected in Isparta-Turkey from healthy individuals and individuals suffering from different urinary disorders.

    Science.gov (United States)

    Ozçelik, N; Koşar, A; Soysal, D

    2001-04-08

    Ochratoxin A (OA) is a nephrotoxic fungal metabolite (mycotoxin) occurring in foodstuffs. The compound is causally associated with mycotoxin porcine nephropathy, a disease comparable with a human kidney disease called endemic nephropathy. In this paper OA levels in the human serum samples collected from healthy individuals and individuals suffering from different urinary disorders in Isparta-Turkey are presented. OA was measured in serum samples of 40 healthy people and a total of 93 patients with different kinds of urinary disorders. Four different kinds of urinary disorders were represented: chronic renal failure treated by hemodialysis (35), chronic renal failure treated by peritoneal dialysis (28), patients with bladder cancer (15), patients with renal stones (15). Analysis of OA in human blood samples was performed using an analytical method based on the measurement of fluorescence spectra. The mean concentration of OA in the healthy group was 0.4 +/- 0.28 ng/ml. The highest mean concentration was found in the group of patients treated by hemodialysis, 2.1+/- 1.2 ng/ml. The mean concentrations of the toxin in all patients groups were higher compared to the control group. Also, a significant difference was found between the mean concentrations of the groups of patients treated by dialysis (hemodialysis or peritoneal dialysis) and of the patients with renal stones or bladder cancer, only with the exception of the difference between peritoneal dialysis and renal stones group. No other significant differences were found when comparing the two groups. The findings indicate that OA may have a role in the human urinary pathology considered herein. A higher level of OA in dialysis groups compared to the control, renal stones and bladder cancer groups could probably be explained by the reduced glomerular filtration rate of these patients.

  19. Bovine serum albumin-Cu(II) hybrid nanoflowers: An effective adsorbent for solid phase extraction and slurry sampling flame atomic absorption spectrometric analysis of cadmium and lead in water, hair, food and cigarette samples

    Energy Technology Data Exchange (ETDEWEB)

    Yilmaz, Erkan [Department of Chemistry, Faculty of Sciences, Erciyes University, Kayseri 38039 (Turkey); Ocsoy, Ismail [Department of Analytical Chemistry, Faculty of Pharmacy, Erciyes University, Kayseri 38039 (Turkey); Nanotechnology Research Center (ERNAM), Erciyes University, Kayseri 38039 (Turkey); Ozdemir, Nalan [Department of Chemistry, Faculty of Sciences, Erciyes University, Kayseri 38039 (Turkey); Soylak, Mustafa, E-mail: soylak@erciyes.edu.tr [Department of Chemistry, Faculty of Sciences, Erciyes University, Kayseri 38039 (Turkey)

    2016-02-04

    Herein, the synthesis of bovine serum albumin-Cu(II) hybrid nanoflowers (BSA-NFs) through the building blocks of bovine serum albumin (BSA) and copper(II) ions in phosphate buffered saline (PBS) and their use as adsorbent for cadmium and lead ions are reported. The BSA-NFs, for the first time, were efficiently utilized as novel adsorbent for solid phase extraction (SPE) of cadmium and lead ions in water, food, cigarette and hair samples. The method is based on the separation and pre-concentration of Cd(II) and Pb(II) by BSA-NFs prior to determination by slurry analysis via flame atomic absorption spectrometry (FAAS). The analytes were adsorbed on BSA-NFs under the vortex mixing and then the ion-loaded slurry was separated and directly introduced into the flame AAS nebulizer by using a hand-made micro sample introduction system to eliminate a number of drawbacks. The effects of analytical key parameters, such as pH, amount of BSA-NFs, vortexing time, sample volume, and matrix effect of foreign ions on adsorbing of Cd(II) and Pb(II) were systematically investigated and optimized. The limits of detection (LODs) for Cd(II) and Pb(II) were calculated as 0.37 μg L{sup −1} and 8.8 μg L{sup −1}, respectively. The relative standard deviation percentages (RSDs) (N = 5) for Cd(II) and Pb(II) were 7.2%, and 5.0%, respectively. The accuracy of the developed procedure was validated by the analysis of certified reference materials (TMDA-53.3 Fortified Water, TMDA-70 Fortified Water, SPS-WW2 Waste Water, NCSDC-73349 Bush Branches and Leaves) and by addition/recovery analysis. The quantitative recoveries were obtained for the analysis of certified reference materials and addition/recovery tests. The method was successfully applied to the analysis of cadmium and lead in water, food, cigarette and hair samples. - Highlights: • The synthesis of bovine serum albumin-Cu(II) hybrid nanoflowers is reported. • The nanoflowers were utilized for solid phase microextraction of

  20. Monitoring of cefepime in human serum and plasma by micellar electrokinetic capillary chromatography: Improvement of sample preparation and validation by liquid chromatography coupled to mass spectrometry.

    Science.gov (United States)

    Šestáková, Nela; Theurillat, Regula; Sendi, Parham; Thormann, Wolfgang

    2017-04-01

    Cefepime monitoring in deproteinized human serum and plasma by micellar electrokinetic capillary chromatography and liquid chromatography coupled to mass spectrometry in presence of other drugs is reported. For micellar electrokinetic capillary chromatography, sample preparation comprised dodecylsulfate protein precipitation at pH 4.5 using an increased buffer concentration compared to that of a previous assay and removal of hydrophobic compounds with dichloromethane. This provided robust conditions for cefepime analysis in the presence of sulfamethoxazole and thus enabled its determination in samples of patients that receive cotrimoxazole. The liquid chromatography assay is based upon use of a column with a pentafluorophenyl-propyl modified and multiendcapped stationary phase and the coupling to electrospray ionization with a single quadrupole detector. The performances of both assays with multilevel internal calibration were assessed with calibration and control samples and both assays were determined to be robust. Cefepime levels monitored by micellar electrokinetic capillary chromatography in samples from patients that were treated with cefepime only and with cefepime and cotrimoxazole were found to compare well with those obtained by liquid chromatography coupled to mass spectrometry. Cefepime drug levels determined by micellar electrokinetic capillary chromatography could thereby be validated. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Simultaneous determination of 12 vitamin D compounds in human serum using online sample preparation and liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Abu Kassim, Nur Sofiah; Shaw, Paul Nicholas; Hewavitharana, Amitha K

    2018-01-19

    The development and validation of a method to simultaneously quantify 12 vitamin D compounds in human serum by LC-MS/MS is described. The main challenge was that of extracting and chromatographing vitamin D compounds with a range of polarities, both lipophilic and hydrophilic, in a single analytical procedure. The extractions of all 12 vitamin D compounds were achieved by an optimised protein precipitation method using acetonitrile as the precipitant, and the separation was accomplished by using a pentafluorophenyl (PFP) column. The sensitivity was increased by minimising matrix effects in MS detector rather than using a lengthy derivatisation procedure; an online solid phase extraction (SPE) using a PFP guard column was used for cleanup. Detection limits for all compounds were in the picomole range when using a 500μL sample volume. Recovery percentages ranged from 92% to 99%. LC-MS/MS resolution of all 12 vitamin D compounds, including the chromatographic separation of 25(OH)D3 from the isomer 3-epi-25(OH)D3 was achieved. Stable isotope labelled vitamin D compounds were used as internal standards for the quantification of all 12 vitamin D compounds. This is a simple yet accurate, selective, and sensitive method for the quantification of 12 major vitamin D compounds, including the sulfated forms, in human serum. The method is sufficiently robust to offer potential for use in routine analysis in a pathology laboratory setting. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Development of a fast sample treatment for the analysis of free and bonded sterols in human serum by LC-MS.

    Science.gov (United States)

    Mendiara, Isabel; Domeño, Celia; Nerín, Cristina

    2012-12-01

    The analysis of sterols in biological fluids allows the clinical study of cholesterol related diseases. This research is focused on reducing the sample processing time of the determination of free and bonded sterols in human serum. Ten sterols were studied: cholesterol precursors (desmosterol, lanosterol, and cholestanol); phytosterols (stigmasterol, campesterol, sitosterol, and sitostanol) and oxysterols (7-α-hydroxy-4-cholesten-3-one, 24-hydroxycholesterol, and 27-hydroxycholesterol). Ultrasound assistance was used to diminish the reaction time during the alkaline hydrolysis for determining total sterols. Different retention mechanisms of solid-phase extraction were compared, two reversed-phase sorbents DSC-18 and polymeric Oasis-HLB and a novel zirconia-coated silica phase. DSC-18 and zirconia-coated silica were the most suitable sorbents to analyze these metabolites. The resulting extracts were analyzed by liquid chromatography coupled to mass spectrometry. The analytical parameters were determined and better values were observed with DSC-18 cartridges for most sterols. LOQ were in the low ng/mL level. Recoveries were in the range 85-99%. Average intermediate precision was 15%. Accuracy for both cartridges was more than 92%. Zirconia-coated silica showed better performance for the oxysterols, with recoveries around 90%. The procedure allows the determination of free and bonded sterol precursors, phytosterols, and oxysterols in human serum. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Serum Samples From Middle-aged Adults Vaccinated Annually with Seasonal Influenza Vaccines Cross-neutralize Some Potential Pandemic Influenza Viruses.

    Science.gov (United States)

    Wang, Wei; Alvarado-Facundo, Esmeralda; Chen, Qiong; Anderson, Christine M; Scott, Dorothy; Vassell, Russell; Weiss, Carol D

    2016-02-01

    We examined serum samples from adults ages 48-64 who received multiple seasonal influenza vaccines from 2004 to 2009 for cross-neutralizing antibodies to potential pandemic strains. Using pseudoviruses bearing various hemagglutinins (HA-pseudoviruses), we found serum neutralization titers (≥160) in 100% against A/Japan/305/1957 (H2N2), 53% against A/Hong Kong/1073/99 (H9N2), 56% against the H3N2 variant A/Indiana/08/11 (H3N2v), 11% against A/Hong Kong/G9/97 (H9N2), and 36% A/chicken/Hong Kong/SF4/01 (H6N1). None had titers >160 to A/Shanghai/2/13 (H7N9) or A/Netherlands/219/03 (H7N7). Thirty-six percent to 0% had neutralization titers to various H5N1 strains. Titers to H9, H6, and H5 HA-pseudoviruses correlated with each other, but not with H3N2v, suggesting group-specific cross-neutralization. Published by Oxford University Press for the Infectious Diseases Society of America 2015. This work is written by (a) US Government employee(s) and is in the public domain in the US.

  4. Relative abundance of hard tick on reared cattle (Family: Bovidae ...

    African Journals Online (AJOL)

    A survey was carried out among cattle reared in Idah LGA of Kogi State, for tick infestations for a period of four months (May – August, 2009). A total of 294 cattle were sampled, 181 were infested with three species of hard ticks (Family: Ixodidae), comprising of Amblyomma variegatum, Boophilus decoloratus, and ...

  5. Field study on nematode resistance in Nelore-breed cattle

    NARCIS (Netherlands)

    Bricarello, P A; Zaros, L G; Coutinho, L L; Rocha, R A; Kooyman, F N J|info:eu-repo/dai/nl/331317788; De Vries, E|info:eu-repo/dai/nl/074757695; Gonçalves, J R S; Lima, L G; Pires, A V; Amarante, A F T

    2007-01-01

    The present study evaluated Nelore cattle with different degrees of resistance to natural infections by gastrointestinal nematodes. One hundred weaned male cattle, 11-12 months of age, were kept on the same pasture and evaluated from October 2003 to February 2004. Faecal and blood samples were

  6. Characterization and Strains of Mycobacteria Isolated from Cattle ...

    African Journals Online (AJOL)

    This work is a pilot study looking kind conducted from March to October 2011 and to determine the frequency of M. bovis strains isolated from cattle carcasses in ... The study also helped to establish the 16 samples positive culture of Mycobacterium bovis distribution of M. bovis based on race, sex, age of cattle and make ...

  7. A survey of traditional beef cattle farmers in Southern Botswana ...

    African Journals Online (AJOL)

    Most of the sampled farmers (56%) used artificial insemination (AI) to improve the genetic quality of their cattle. The constraints, which prevented farmers from using AI were long distances (over 60 km) between cattle post and AI camp centre, limited carrying capacity of the AI camps, small numbers of cows permitted per ...

  8. A survey of small-scale cattle farming systems in the North West ...

    African Journals Online (AJOL)

    A questionnaire-based survey was conducted among a purposive sample of 60 small-scale cattle farmers in the Mopeane Rustenburg district, all of whom owned five or more head of cattle. Results showed that small-scale cattle farming is a part-time activity for most farmers in this district, and a way of investing or saving ...

  9. A novel SNP of Chemerin gene in cattle and its association with ...

    African Journals Online (AJOL)

    User

    2012-04-26

    Apr 26, 2012 ... SNP 2692C>T. It was shown that associations do exist between chemerin gene and carcass and meat quality traits. As a result of the small sample size of this study, it is ..... HW, Hardy-Weinberg equilibrium; QC, Qinchuan cattle breed; CYR, Caoyuan red cattle breed; JR, Jiaxian red cattle breed; LX, Luxi.

  10. Impact of cattle congregation sites on soil nutrients and soil compaction

    Science.gov (United States)

    This study determined the impact of grazing cattle on the changes in soil quality around and beneath cattle congregation sites (mineral feeders, water troughs, and shades). Baseline soil samples around and beneath three congregations sites in established (>10 yr) grazed beef cattle pastures at the U...

  11. Seroprevalence of leptospiral infection in feline population in urban and dairy cattle herds in Mashhad, Iran.

    Science.gov (United States)

    Talebkhan Garoussi, Massoud; Mehravaran, Mohsen; Abdollahpour, Gholamreza; Khoshnegah, Javad

    2015-01-01

    The importance of cats in the Leptospira epidemiology is due to the possibility of transferring leptospirosis to wild and domesticated animals. The purpose of this survey was to determine the prevalence of Leptospira infection in shorthair cats in different location of Mashhad, Iran. Totally, 147 blood samples were taken from 42 (28.57%), 52 (35.37%) and 53 (36.05%) households, stray and cats which lived in industrial dairy cattle herds of Mashhad, Iran, respectively. Sera were tested with seven live Leptospira antigens using microscopic agglutination test (MAT). Sera with 50.00% agglutination at the dilution of ≥ 1/100 were considered as positive samples. Agglutination at dilutions of < 1/100 considered as suspected to Leptospira infection. Overall, 19 (12.92%) out of 147 cats showed reaction in MAT. The seroprevalence at a titer ≥ 1:100 and < 1:100 were 10 (6.80%) and 9 (6.12%), respectively. Serum samples showed positive reaction against Leptospira intterogans hardjo (no = 10; 52.63%), pomona (no = 5; 26.31%) and icterohaemorrhagiae (no = 4; 21.05%). Eight cats (42.10%) belong to dairy cattle herds had the most infection only by L. I. hardjo with 1:200 titer. There were no significant differences among the weight' age and sex of infected cats. However, there were significant differences between the infected cats in dairy cattle herds and the cats in the urban area (p < 0.05). It is concluded that cats can be infected by Leptospira spp. especially in commercial dairy cattle herds. Cats can be considered as a sanitation hazards in the area for this zoonotic disease.

  12. A new supramolecular based liquid solid microextraction method for preconcentration and determination of trace bismuth in human blood serum and hair samples by electrothermal atomic absorption spectrometry.

    Science.gov (United States)

    Kahe, Hadi; Chamsaz, Mahmoud

    2016-11-01

    A simple and reliable supramolecule-aggregated liquid solid microextraction method is described for preconcentration and determination of trace amounts of bismuth in water as well as human blood serum and hair samples. Catanionic microstructures of cetyltrimethylammonium bromide (CTAB) and sodium dodecyl sulfate (SDS) surfactants, dissolved in deionized water/propanol, are used as a green solvent to extract bismuth (III)-diethyldithiocarbamate complexes by dispersive microextraction methodology. The extracted solid phase is easily removed and dissolved in 50 μL propanol for subsequent measurement by electrothermal atomic absorption spectrometry (ET-AAS). The procedure benefits the merits of supramolecule aggregates' properties and dispersive microextraction technique using water as the main component of disperser solvent, leading to direct interaction with analyte. Phase separation behavior of extraction solvent and different parameters influencing the extraction efficiency of bismuth ion such as salt concentration, pH, centrifugation time, amount of chelating agent, SDS:CTAB mole ratio, and solvent amounts were thoroughly optimized. Under the optimal experimental conditions, the calibration curve was linear in the range of 0.3-6 μg L-1 Bi (III) with a limit of detection (LOD) of 0.16 μg L-1 (S/N = 3). The relative standard deviations (RSD) of determination were obtained to be 5.1 and 6.2 % for 1 and 3 μg L-1 of Bi (III), respectively. The developed method was successfully applied as a sensitive and accurate technique for determination of bismuth ion in human blood serum, hair samples, and a certified reference material.

  13. Automated isotope dilution liquid chromatography-tandem mass spectrometry with on-line dilution and solid phase extraction for the measurement of cortisol in human serum sample.

    Science.gov (United States)

    Kawaguchi, Migaku; Eyama, Sakae; Takatsu, Akiko

    2014-08-05

    A candidate reference measurement procedure involving automated isotope dilution coupled with liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) with on-line dilution and solid phase extraction (SPE) has been developed and critically evaluated. We constructed the LC-MS/MS with on-line dilution and SPE system. An isotopically labelled internal standard, cortisol-d4, was added to serum sample. After equilibration, the methanol was added to the sample, and deproteination was performed. Then, the sample was applied to the LC-MS/MS system. The limit of detection (LOD) and limit of quantification (LOQ) were 0.2 and 1ngg(-1), respectively. Excellent precision was obtained with within-day variation (RSD) of 1.9% for ID-LC-MS/MS analysis (n=6). This method, which demonstrates simple, easy, good accuracy, high precision, and is free from interferences from structural analogues, qualifies as a reference measurement procedure. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Simplifying sample preparation using fabric phase sorptive extraction technique for the determination of benzodiazepines in blood serum by high-performance liquid chromatography.

    Science.gov (United States)

    Samanidou, Victoria; Kaltzi, Ioanna; Kabir, Abuzar; Furton, Kenneth G

    2016-06-01

    Fabric phase sorptive extraction (FPSE), a recently introduced novel sample preparation technology, has been evaluated for the extraction of benzodiazepines from human blood serum. FPSE utilizes a flexible fabric surface as the substrate platform for creating sol-gel hybrid organic-inorganic sorbent coatings. FPSE media can be introduced directly into the sample containing the target analyte(s), requiring no need for prior sample pretreatment or clean-up. Benzodiazepines were selected as model analytes because they represent one of the most widely used therapeutic drugs in psychiatry and are also amongst the most frequently encountered drugs in forensic toxicology. The chromatographic separation of target analytes was performed on a LiChroCART-LiChrospher®100 RP-18e (5 µm, 250 × 4 mm) analytical column, operated at room temperature. Ternary gradient elution was applied with a mobile phase that consisted of acetonitrile, methanol and ammonium acetate (0.05 M), which was delivered at a flow rate of 1.0 mL/min. Diode array detection was performed with monitoring at 240 nm. FPSE was performed using cellulose fabric extraction media coated with sol-gel poly(ethylene glycol) (sol-gel PEG). Absolute recovery values in the equilibrium state for the examined benzodiazepines were found to be 27% for bromazepam, 63% for lorazepam, 42 % for diazepam and 39% for alprazolam. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

  15. β-Cyclodextrin anchoring onto pericarpium granati-derived magnetic mesoporous carbon for selective capture of lopid in human serum and pharmaceutical wastewater samples.

    Science.gov (United States)

    Liu, Rui-Lin; Zhang, Zhi-Qi; Jing, Wang-Hui; Wang, Lu; Luo, Zhi-Min; Chang, Rui-Miao; Zeng, Ai-Guo; Du, Wei; Chang, Chun; Fu, Qiang

    2016-05-01

    Functionalized magnetic carbonaceous nanomaterials, which are important materials with many practical and research applications in biomedical, pharmaceutical and biological fields, have recently attracted much attention. In this study, a magnetic mesoporous carbon coated with β-cyclodextrin (MMC@β-CD) was synthesized for the first time from natural pericarpium granati (PG). The as-obtained MMC@β-CD has high surface areas (203 m(2)g(-1)), large pore volumes (0.16 cm(3)g(-1)), relatively broad mesoporous sizes (6.8 nm) and a high saturation magnetization of 26.2 emu g(-1), which is sufficient for magnetic separation by an external magnetic field. The MMC@β-CD was used as an innovative adsorbent for magnetic solid-phase extraction of lopid via host-guest interaction prior to spectrofluorometric analysis. The proposed method was successfully applied to analyze lopid in human serum and pharmaceutical wastewater samples with recoveries in the range of 85.0-103.5% for the spiked samples. Overall, this work not only provides an inexpensive and eco-friendly method to fabricate MMC@β-CD (or MMC) from PG, but also develops a highly selective approach for capture of lopid in biological samples and environmental substances. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Genetic diversity of bovine viral diarrhea virus in cattle from Mexico.

    Science.gov (United States)

    Gómez-Romero, Ninnet; Basurto-Alcántara, Francisco J; Verdugo-Rodríguez, Antonio; Bauermann, Fernando V; Ridpath, Julia F

    2017-05-01

    Bovine viral diarrhea virus (BVDV) infects cattle populations worldwide, causing significant economic losses though its impact on animal health. Previous studies have reported the prevalence of BVDV species and subgenotypes in cattle from the United States and Canada. We investigated the genetic diversity of BVDV strains detected in bovine serum samples from 6 different Mexican regions. Sixty-two BVDV isolates from Mexico were genetically typed based on comparison of sequences from the 5' untranslated region (5'-UTR) of the viral genome. Phylogenetic reconstruction indicated that 60 of the samples belonged to the BVDV-1 genotype and 2 to the BVDV-2 genotype. Comparison of partial 5'-UTR sequences clustered 49 samples within BVDV-1c, 8 samples within BVDV-1a, 3 samples within BVDV-1b, and 2 samples clustered with the BVDV-2a subgenotypes. Our study, combined with information previously published on BVDV field strain diversity in the United States and Canada, benefits the development of effective detection assays, vaccines, and control programs for North America.

  17. Identification of cattle, buffaloes and rodents as reservoir animals of Leptospira in the District of Gampaha, Sri Lanka.

    Science.gov (United States)

    Denipitiya, D T H; Chandrasekharan, N V; Abeyewickreme, W; Hartskeerl, R A; Hapugoda, M D

    2017-03-23

    Leptospirosis is an important emerging infectious disease in Sri Lanka. Rats are the most important reservoir of Leptospira but domestic and wild mammals may also act as important maintenance or accidental hosts. In Sri Lanka, knowledge of reservoir animals of leptospires is poor. The objective of this study was to identify potential reservoir animals of Leptospira in the District of Gampaha, Sri Lanka. Blood and kidney samples were collected from 38 rodents and mid-stream urine samples were randomly collected from 45 cattle and five buffaloes in the District of Gampaha. Kidney and urine samples were tested by real-time polymerase chain reaction (PCR) and serum samples were tested by the microscopic agglutination test (MAT). Of the 38 rodent kidney samples, 11% (4/38) were positive by real-time PCR. The prevalence of leptospiral carriage was 11% (3/26) and 8% (1/12) in female and male rodents, respectively. Three rodent serum samples were positive by MAT. Of the 50 cattle/buffalo urine samples tested, 10% (5/50) were positive by real-time PCR. The prevalence of leptospiral carriage was 9% (4/45) and 20% (1/5) in cattle and buffaloes, respectively. Results of PCR and MAT showed that Leptospira were present in a significant proportion of the rodents and farm animals tested in this study and suggest that these (semi-) domestic animals form an infection reservoir for Leptospira. Therefore, there is a potential zoonotic risk to public health, most notably to farmers in this area.

  18. Bestimmung der blut/serum-verhältnisse verschiedener forensisch relevanter analyten in authentischen proben [ Determination of blood/serum ratios of different forensically relevant analytes in authentic samples

    NARCIS (Netherlands)

    Jantos, R.; Schuhmacher, M.; Veldstra, J.L.; Bosker, W.M.; Klöpping-Ketelaars, I.; Touliou, K.; Sardi, G.M.; Brookhuis, K.A.; Ramaekers, J.G.; Mattern, R.; Skopp, G.

    2011-01-01

    For forensic toxicological investigations only whole blood, but no serum is often available. Pharmacokinetic data are helpful for interpreting the results, but most of these studies indicate serum or plasma concentrations. In order to obtain reliable conversion factors which also take intersubject

  19. Seroepidemiological investigation of foot-and-mouth disease virus serotypes in cattle around Lake Mburo National Park in South-Western Uganda

    DEFF Research Database (Denmark)

    Mwiine, Frank Norbert; Ayebazibwe, Chrisostom; Alexandersen, Søren

    2010-01-01

    Foot-and-mouth disease (FMD) outbreaks in cattle occur annually in Uganda. In this study the authors investigated antibodies against FMD virus (FMDV) in cattle in surrounding areas of Lake Mburo National Park in South-western Uganda. Two hundred and eleven serum samples from 23 cattle herds were...... in SPBEs were identified in 61% (19/31), 33% (5/15), 6%7 (20/30), 37% (10/27) and 12% (4/33) of the investigated samples for serotypes O, A, SAT 1, SAT 2 and SAT 3, respectively. This study indicates that most of the FMD outbreaks in the cattle herds in the investigated area were probably caused by FMDV...... serotype O, A and/ or SAT-serotype(s). It also shows that the usage of non-purified, multivalent vaccines in Uganda obscures the serological diagnosis of FMDV outbreaks, and that the sampling strategy needs to be improved. Finally, it emphasizes the importance of isolation and characterization of FMD...

  20. The diversity of BVDV subgenotypes in a vaccinated dairy cattle herd in Brazil.

    Science.gov (United States)

    Otonel, Rodrigo A A; Alfieri, Alice F; Dezen, Stelamaris; Lunardi, Michele; Headley, Selwyn A; Alfieri, Amauri A

    2014-01-01

    Bovine viral diarrhoea virus (BVDV) is an important pathogen of cattle that occurs worldwide with substantial economic impact on beef and dairy industries. The aim of this study was to describe the diversity of BVDV subgenotypes in persistently infected (PI) animals identified in a highly productive, regularly vaccinated, dairy cattle herd presenting with reproductive failure. Serum samples were collected from all animals within the herd (n = 692) and used to detect the presence of BVDV RNA. Using reverse transcription polymerase chain reaction assay, 29 cows were identified as transiently infected, three animals (two cows and one calf) as persistently infected, and one calf as putative BVDV PI animal. The sequences of 5'UTR and/or N(pro) gene of BVDV used in phylogenetic analyses revealed that the three PI animals were infected by three different BVDV subgenotypes (BVDV-1a, BVDV-1b, and BVDV-1d). These results demonstrated that in an open dairy cattle herd, regular vaccination against BVDV by itself is not able to prevent viral circulation in the herd. Furthermore, depending on the frequency of the acquisition of heifers and/or cows for replacement, several BVDV subgenotypes may co-exist simultaneously in the same herd.

  1. Ultra-sensitive detection of prion protein fibrils by flow cytometry in blood from cattle affected with bovine spongiform encephalopathy

    Directory of Open Access Journals (Sweden)

    Maas Elke

    2005-10-01

    Full Text Available Abstract Background The definite diagnosis of prion diseases such as Creutzfeldt-Jakob disease (CJD in humans or bovine spongiform encephalopathy (BSE in cattle currently relies on the post mortem detection of the pathological form of the prion protein (PrPSc in brain tissue. Infectivity studies indicate that PrPSc may also be present in body fluids, even at presymptomatic stages of the disease, albeit at concentrations well below the detection limits of currently available analytical methods. Results We developed a highly sensitive method for detecting prion protein aggregates that takes advantage of kinetic differences between seeded and unseeded polymerization of prion protein monomers. Detection of the aggregates was carried out by flow cytometry. In the presence of prion seeds, the association of labelled recombinant PrP monomers in plasma and serum proceeds much more efficiently than in the absence of seeds. In a diagnostic model system, synthetic PrP aggregates were detected down to a concentration of approximately 10-8 nM [0.24 fg/ml]. A specific signal was detected in six out of six available serum samples from BSE-positive cattle. Conclusion We have developed a method based on seed-dependent PrP fibril formation that shows promising results in differentiating a small number of BSE-positive serum samples from healthy controls. This method may provide the basis for an ante mortem diagnostic test for prion diseases.

  2. Validation of a reference method for total cholesterol measurement in human serum and assignation of reference values to proficiency testing samples.

    Science.gov (United States)

    Heuillet, Maud; Lalere, Beatrice; Peignaux, Maryline; De Graeve, Jacques; Vaslin-Reimann, Sophie; Pais De Barros, Jean-Paul; Gambert, Philippe; Duvillard, Laurence; Delatour, Vincent

    2013-03-01

    Our objective was to develop a reference method to measure total cholesterol in human serum, in order to assign values and assess the accuracy of field methods in French clinical laboratories. A reference method based on gas chromatography coupled with mass spectrometry and isotope dilution (GC-IDMS) was developed and validated. It was then used to assign reference values to five frozen serum samples from voluntary proficiency testing schemes gathering 170 French clinical laboratories. Three peer groups were defined and bias against the reference method target value was calculated. Accuracy of the reference method was assessed against NIST SRM 1951b. Bias of the reference method was less than 0.5% and imprecision was less than 1.0%. Our study indicated that field methods tended to overestimate total cholesterol concentration, mean bias being +5.02% ± 1.02%. The most popular methods (phenolic chromogen with spectrophotometric detection, 80% of participants) exhibited the highest bias (peer group mean bias: +5.51 ± 1.24%). Neither these methods nor those using a non-phenolic chromogen with reflectometric detection (10% of participants, peer group mean bias: +4.20 ± 1.44%) met NCEP recommendations according to which bias should be less than 3%. Only the methods using a non phenolic chromogen with a spectrophotometric detection met these recommendations (10% of participants, peer group mean bias: +1.39 ± 2.75%). As all three peer groups provided positively biased results, the consensus mean usually used to assess the trueness of routine methods is biased as well, which results in an erroneous estimation of method bias. Therefore, this study highlights the value added by reference method target values to assess trueness of field methods and monitor performance of clinical laboratories. Copyright © 2012 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  3. GENETIC CHARACTERIZATION OF ROMANIAN CATTLE BREEDS USING BIOCHEMICAL MARKERS

    Directory of Open Access Journals (Sweden)

    REBEDEA MARIANA

    2007-01-01

    Full Text Available The paper presents a genetic characterization of cattle breeds in Romania based onbiochemical markers in the blood and the milk. The surveyed breeds are: RomanianBlack Spotted Cattle (BNR, Romanian Spotted Cattle (BR, Romanian Brown (Band Romanian Steppe, and the markers identified are represented by some proteins,serum transferrin (Tf, serum albumins (Al, hemoglobin (Hb respectively-from theblood and beta-lactoglobulin (βLg-from the milk. In order to determine thegenotypes in the studied populations electrophoresis was used in three differentvariations, depending on the type of the protein, and the migration substrates usedwere starch and polyacrylamide. The identified genetic structures in the individualsfrom the surveyed breeds allowed their genetic characterization based on gene andgenotype frequencies, as well as using these data in establishing the identity andpaternity of the individuals in the surveyed breeds.

  4. Serological Investigation And Interpreting Serum Chemistry Profile ...

    African Journals Online (AJOL)

    From the above number of samples 20 serum samples only are used as infected group and 20 serum samples from clinically healthy animals considered as control group. Serum samples were used to establish the concentration of total protein, albumin, glucose, total cholesterol, calcium and phosphorus. Serum nitrate ...

  5. Trace analysis of mefenamic acid in human serum and pharmaceutical wastewater samples after pre-concentration with Ni–Al layered double hydroxide nano-particles

    Directory of Open Access Journals (Sweden)

    Hossein Abdolmohammad-Zadeh

    2014-10-01

    Full Text Available In this work, the nickel–aluminum layered double hydroxide (Ni–Al LDH with nitrate interlayer anion was synthesized and used as a solid phase extraction sorbent for the selective separation and pre-concentration of mefenamic acid prior to quantification by UV detection at λmax=286 nm. Extraction procedure is based on the adsorption of mefenamate anions on the Ni–Al(NO3− LDH and/or their exchange with LDH interlayer NO3− anions. The effects of several parameters such as cations and interlayer anions type in LDH structure, pH, sample flow rate, elution conditions, amount of nano-sorbent and co-existing ions on the extraction were investigated and optimized. Under the optimum conditions, the calibration graph was linear within the range of 2–1000 µg/L with a correlation coefficient of 0.9995. The limit of detection and relative standard deviation were 0.6 µg/L and 0.84% (30 µg/L, n=6, respectively. The presented method was successfully applied to determine of mefenamic acid in human serum and pharmaceutical wastewater samples.

  6. Relationship between serum uric acid and electrocardiographic alterations in a large sample of general population: data from the Brisighella Heart Study.

    Science.gov (United States)

    Cicero, Arrigo F G; Rosticci, Martina; Reggi, Alessandra; Derosa, Giuseppe; Parini, Angelo; Grandi, Elisa; D'Addato, Sergio; Borghi, Claudio

    2015-06-01

    Serum uric acid (SUA) may contribute to the increased cardiovascular damage through direct injury to the endothelium and alteration of cardiovascular function. To evaluate the association of SUA with the presence of the most recurrent electrographic alterations and with the length of the main ECG intervals in a large sample of general population. For this study, on the database of the Brisighella Heart Study, we evaluated the available data of 790 men and 849 women, excluding subjects affected by gout or taking antihyperuricemic agents, those taking drug increasing the QT interval and those using beta-blockers or non-dihydropyridine calcium channel blockers at the moment of the ECG registration. Multiple ascending stepwise regression analyses were carried out to determine the independent predictors of the predefined ECG alterations. The prevalence of predefined ECG alterations was comparable between genders, with the exception of sinus bradicardia, left-anterior fascicular block, atrio-ventricular blocks and left ventricular hypertrophy (LVH), which appeared to be more frequent in men. The multivariate analysis revealed that SUA was associated to ischaemic alterations, LVH, sinus tachycardia and tachyarrhytmias. Age was associated to all evaluated ECG alterations beyond sinus tachycardia and LVH. Male sex was associated to sinus bradicardia, atrio-ventricular blocks, anterior-left fascicular block and LVH. Blood pressure was associated to different ECG alterations, but with clinically relevant OR with ischaemic alterations and LVH. SUA level is related the prevalence of both organic and rhythm ECG alterations in a wide sample of general population.

  7. A retrospective study of 1,098 blood samples with anemia from adult cats: frequency, classification, and association with serum creatinine concentration.

    Science.gov (United States)

    Furman, E; Leidinger, E; Hooijberg, E H; Bauer, N; Beddies, G; Moritz, A

    2014-01-01

    Frequency and classification of anemia in terms of regeneration status and erythrocyte indices are not well described in cats. To determine frequency and regenerative status of anemia in samples from adult cats, to assess the sensitivity and specificity of macrocytosis and hypochromasia for detecting regenerative anemia (RA), and to evaluate the association of anemia with increased serum creatinine concentration (SC). Laboratory records from 30,503 blood samples from cats (2003-2011). Clinicopathologic data reviewed retrospectively. Anemia defined as hematocrit (Ht) ≤27%, red blood cell count (RBC) ≤5.5 × 10(6)/μL and hemoglobin (Hb) ≤9.0 g/dL. RA defined by manual absolute reticulocyte count >50 × 10(3)/μL. Macrocytosis was defined as mean corpuscular volume (MCV) >55 fL and hypochromasia as mean corpuscular hemoglobin concentration (MCHC) anemia, 633 of 1,098 (57.7%) classified as nonregenerative (NRA) and 465 of 1,098 (42.3%) as regenerative. RBC, Ht, and Hb were significantly lower in the RA compared to NRA group (P Anemia was more severe in cats with RA. Erythrocyte indices were not sensitive indicators of RA. Copyright © 2014 by the American College of Veterinary Internal Medicine.

  8. Detection of lipomannan in cattle infected with bovine tuberculosis

    Science.gov (United States)

    Early and rapid detection of bovine tuberculosis (bTB) is critical to controlling the spread of this disease in cattle and other animals. In this study, we demonstrate the development of an immunoassay for the direct detection of the bovine bTB biomarker, lipomannan (LM) in serum using a waveguide-...

  9. Mineral element status of soils, native pastures and cattle blood ...

    African Journals Online (AJOL)

    Based on the information obtained from soil, native pasture and serum assays, low levels of Zn, P, Cu and Mg in soil and native pasture could potentially limit livestock production in the study area. Keywords: animal nutrition; communal grazing; Horro cattle; seasonal changes; minerals. African Journal of Range & Forage ...

  10. Evaluation of hair cortisol in beef cattle of divergent temperaments

    Science.gov (United States)

    The objective of this research project was to evaluate the relationships among hair and serum cortisol concentrations and cattle disposition. Spring born (n = 101) crossbred beef heifers (7 to 8 mo. of age) were evaluated for temperament preweaning and at weaning by pen score (PS; 1 = calm and 5 = e...

  11. Sero-epidemiology of Breda virus in cattle using ELISA

    NARCIS (Netherlands)

    Horzinek, M.C.; Koopmans, M.P.; Boom, U. van den; Woode, G.N.

    1989-01-01

    Two direct blocking enzyme linked immunosorbent assays (ELISA) for the detection of antibodies to Breda virus in sera of cattle were compared. An ELISA with consecutive addition of antigen and test serum to an antibody-coated plate gave higher positive: negative absorbence ratios than an ELISA in

  12. Bovine serum albumin-Cu(II) hybrid nanoflowers: An effective adsorbent for solid phase extraction and slurry sampling flame atomic absorption spectrometric analysis of cadmium and lead in water, hair, food and cigarette samples.

    Science.gov (United States)

    Yilmaz, Erkan; Ocsoy, Ismail; Ozdemir, Nalan; Soylak, Mustafa

    2016-02-04

    Herein, the synthesis of bovine serum albumin-Cu(II) hybrid nanoflowers (BSA-NFs) through the building blocks of bovine serum albumin (BSA) and copper(II) ions in phosphate buffered saline (PBS) and their use as adsorbent for cadmium and lead ions are reported. The BSA-NFs, for the first time, were efficiently utilized as novel adsorbent for solid phase extraction (SPE) of cadmium and lead ions in water, food, cigarette and hair samples. The method is based on the separation and pre-concentration of Cd(II) and Pb(II) by BSA-NFs prior to determination by slurry analysis via flame atomic absorption spectrometry (FAAS). The analytes were adsorbed on BSA-NFs under the vortex mixing and then the ion-loaded slurry was separated and directly introduced into the flame AAS nebulizer by using a hand-made micro sample introduction system to eliminate a number of drawbacks. The effects of analytical key parameters, such as pH, amount of BSA-NFs, vortexing time, sample volume, and matrix effect of foreign ions on adsorbing of Cd(II) and Pb(II) were systematically investigated and optimized. The limits of detection (LODs) for Cd(II) and Pb(II) were calculated as 0.37 μg L(-)(1) and 8.8 μg L(-)(1), respectively. The relative standard deviation percentages (RSDs) (N = 5) for Cd(II) and Pb(II) were 7.2%, and 5.0%, respectively. The accuracy of the developed procedure was validated by the analysis of certified reference materials (TMDA-53.3 Fortified Water, TMDA-70 Fortified Water, SPS-WW2 Waste Water, NCSDC-73349 Bush Branches and Leaves) and by addition/recovery analysis. The quantitative recoveries were obtained for the analysis of certified reference materials and addition/recovery tests. The method was successfully applied to the analysis of cadmium and lead in water, food, cigarette and hair samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. The Sero-epidemiology of Coxiella burnetii in Humans and Cattle, Western Kenya: Evidence from a Cross-Sectional Study.

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    Nicola A Wardrop

    2016-10-01

    Full Text Available Evidence suggests that the intracellular bacterial pathogen Coxiella burnetii (which causes Q fever is widespread, with a near global distribution. While there has been increasing attention to Q fever epidemiology in high-income settings, a recent systematic review highlighted significant gaps in our understanding of the prevalence, spatial distribution and risk factors for Q fever infection across Africa. This research aimed to provide a One Health assessment of Q fever epidemiology in parts of Western and Nyanza Provinces, Western Kenya, in cattle and humans. A cross-sectional survey was conducted: serum samples from 2049 humans and 955 cattle in 416 homesteads were analysed for C. burnetii antibodies. Questionnaires covering demographic, socio-economic and husbandry information were also administered. These data were linked to environmental datasets based on geographical locations (e.g., land cover. Correlation and spatial-cross correlation analyses were applied to assess the potential link between cattle and human seroprevalence. Multilevel regression analysis was used to assess the relationships between a range of socio-economic, demographic and environmental factors and sero-positivity in both humans and animals. The overall sero-prevalence of C. burnetii was 2.5% in humans and 10.5% in cattle, but we found no evidence of correlation between cattle and human seroprevalence either within households, or when incorporating spatial proximity to other households in the survey. Multilevel modelling indicated the importance of several factors for exposure to the organism. Cattle obtained from market (as opposed to those bred in their homestead and those residing in areas with lower precipitation levels had the highest sero-prevalence. For humans, the youngest age group had the highest odds of seropositivity, variations were observed between ethnic groups, and frequent livestock contact (specifically grazing and dealing with abortion material was

  14. The Sero-epidemiology of Coxiella burnetii in Humans and Cattle, Western Kenya: Evidence from a Cross-Sectional Study.

    Science.gov (United States)

    Wardrop, Nicola A; Thomas, Lian F; Cook, Elizabeth A J; de Glanville, William A; Atkinson, Peter M; Wamae, Claire N; Fèvre, Eric M

    2016-10-01

    Evidence suggests that the intracellular bacterial pathogen Coxiella burnetii (which causes Q fever) is widespread, with a near global distribution. While there has been increasing attention to Q fever epidemiology in high-income settings, a recent systematic review highlighted significant gaps in our understanding of the prevalence, spatial distribution and risk factors for Q fever infection across Africa. This research aimed to provide a One Health assessment of Q fever epidemiology in parts of Western and Nyanza Provinces, Western Kenya, in cattle and humans. A cross-sectional survey was conducted: serum samples from 2049 humans and 955 cattle in 416 homesteads were analysed for C. burnetii antibodies. Questionnaires covering demographic, socio-economic and husbandry information were also administered. These data were linked to environmental datasets based on geographical locations (e.g., land cover). Correlation and spatial-cross correlation analyses were applied to assess the potential link between cattle and human seroprevalence. Multilevel regression analysis was used to assess the relationships between a range of socio-economic, demographic and environmental factors and sero-positivity in both humans and animals. The overall sero-prevalence of C. burnetii was 2.5% in humans and 10.5% in cattle, but we found no evidence of correlation between cattle and human seroprevalence either within households, or when incorporating spatial proximity to other households in the survey. Multilevel modelling indicated the importance of several factors for exposure to the organism. Cattle obtained from market (as opposed to those bred in their homestead) and those residing in areas with lower precipitation levels had the highest sero-prevalence. For humans, the youngest age group had the highest odds of seropositivity, variations were observed between ethnic groups, and frequent livestock contact (specifically grazing and dealing with abortion material) was also a risk

  15. Evaluation of the relationship between some blood metabolites and elements on the ovarian persistent follicle formation in dairy cattle

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    Z Merati

    2011-02-01

    Full Text Available The object of this study was to investigate the effect of some blood metabolites and elements on the ovarian persistent follicle formation in dairy cattle. Blood samples were obtained from 117 lactating cows in two groups of normal and persistent follicle status. Glucose, urea, total protein, phosphorus and betahydroxy butyrate were measured by spectrophotometer method. Estradiol concentration in blood serum was measured by ELISA method. Effects of calving season, parity and milk production at 50-60 days postpartum were studied on persistent follicle status. Status of animal (presence or absence of resistant follicle had significant effect on urea, protein, beta-hydroxybutyrate, phosphorus and estradiol (p

  16. Prevalência de anticorpos contra o vírus da língua azul em bovinos e ovinos do Sudoeste e Sudeste do Rio Grande do Sul Bluetongue virus antibodies in cattle and sheep in Southwest and Southeast regions of Rio Grande do Sul, Brazil

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    J.R.R. Costa

    2006-04-01

    Full Text Available It was studied bluetongue virus antibodies prevalence for sheep and cattle in Southwest and Southeast regions of Rio Grande do Sul State. A total of 2613 serum samples (1272 bovine and 1341 ovine were tested by agar gel immunodiffusion. Eight bovine and two ovine samples were positive meaning a prevalence of 0.63% and 0.15%, respectively. These results show that most of animals in these regions are negative to bluetongue.

  17. Comparison of PRRSV Nucleic Acid and Antibody Detection in Pen-Based Oral Fluid and Individual Serum Samples in Three Different Age Categories of Post-Weaning Pigs from Endemically Infected Farms

    Science.gov (United States)

    De Regge, Nick; Cay, Brigitte

    2016-01-01

    Background Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of an economically important disease in swine. Since it has been shown that PRRSV and PRRSV specific antibodies can be detected in oral fluid, many different aspects have been studied to show that oral fluid could be a worthy alternative diagnostic sample to serum for monitoring and surveillance of this disease. Thorough field evaluations are however missing to convincingly show its usefulness under representative field conditions. Methodology Pen-based oral fluid samples and serum samples from all individual pigs in the corresponding pens were collected from post-weaning pigs of three different age categories in eight endemically PRRSV infected farms and one PRRSV free farm in Belgium. All samples were tested by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and ELISA to detect PRRSV RNA and PRRSV specific antibodies, respectively. Results While the relative specificity of PRRSV detection by qRT-PCR in pen-based oral fluid compared to serum collected from individual pigs was high in all age categories (>90%), the relative sensitivity decreased with the age of the pigs (89, 93 and 10% in 8-12w, 16-20w and 24-28w old pigs, respectively). The latter correlated with a lower percentage of PRRSV positive pigs in serum/pen in the different age categories (55, 29 and 6%, respectively). Irrespective of the age category, pen-based oral fluid samples were always found PCR positive when at least 30% of the individual pigs were positive in serum. PRRSV specific antibody detection in oral fluid by ELISA showed a 100% relative sensitivity to detection in serum since oral fluid samples were always positive as soon as one pig in the pen was positive in serum. On the other hand, two false positive oral fluid samples in 11 pens without serum positive pigs were found, resulting in a relative specificity of 82%. Indications are however present that the oral fluid

  18. Quantitative chiral and achiral determination of ketamine and its metabolites by LC-MS/MS in human serum, urine and fecal samples.

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    Hasan, Mahmoud; Hofstetter, Robert; Fassauer, Georg M; Link, Andreas; Siegmund, Werner; Oswald, Stefan

    2017-05-30

    Ketamine (KET) is a widely used anesthetic drug which is metabolized by CYP450 enzymes to norketamine (n-KET), dehydronorketamine (DHNK), hydroxynorketamine (HNK) and hydroxyketamine (HK). Ketamine is a chiral compound and S-ketamine is known to be the more potent enantiomer. Here, we present the development and validation of three LC-MS/MS assays; the first for the quantification of racemic KET, n-KET and DHNK in human serum, urine and feces; the second for the separation and quantification of the S- and R-enantiomers of KET, n-KET and DHNK, and the third for separation and quantification of 2S,6S-hydroxynorketamine (2S,6S-HNK) and 2R,6R-hydroxynorketamine (2R,6R-HNK) in serum and urine with the ability to separate and detect 10 additional hydroxylated norketamine metabolites of racemic ketamine. Sample preparation was done by liquid-liquid extraction using methyl tert-butyl ether. For achiral determination of KET and its metabolites, an isocratic elution with ammonium acetate (pH 3.8; 5mM) and acetonitrile on a C18 column was performed. For the separation of S- and R-enantiomers of KET, n-KET and DHNK, a gradient elution was applied using a mobile phase of ammonium acetate (pH 7.5; 10mM) and isopropanol on the CHIRAL-AGP ® column. The enantioselective separation of the HNK metabolites was done on the chiral column Lux ® -Amylose-2 with a gradient method using ammonium acetate (pH 9; 5mM) and a mixture of isopropanol and acetonitrile (4:1). The mass spectrometric detection monitored for each analyte 2-3 mass/charge transitions. D4-ketamine and D4-n-KET were used as internal standards. The assays were successfully validated according to current bioanalytical guidelines and applied to a pilot study in one healthy volunteer. Compared to previously published methods, our assays have superior analytical features such as a lower amount of required matrix, faster sample preparation, shorter analytical run time and higher sensitivity (LLOQ up to 0.1ng/ml). Moreover, our

  19. Intervention of genetic flow of the foreign cattle toward diversity of phenotype expressions of local cattle in the District of Banyuwangi

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    MOHAMAD AMIN

    2010-04-01

    Full Text Available Amin M (2010 Intervention of genetic flow of the foreign cattle toward diversity of phenotype expressions of local cattle in the District of Banyuwangi. Biodiversitas 10: 69-74. The aims of the present research are two folds: to know the phenotypic diversity and to reconstruct the cross-breeding pattern of local cattle in Banyuwangi. Based on three sampling areas, it was found that there were 32 phenotypic cattle (10 in the sub districts of Rogojampi, 16 in Tegaldlimo and 6 in Glagah areas. The phenotypic varieties were caused by two factors, namely the flow of genetic intervention of the other local cattle (Bali, Ongole, and Brahman cattle and the artificial insemination program using the semen of Limousine, Simmental, Aberdeen Angus and Santa Gertrudis cattle.

  20. An investigation into an outbreak of Rift Valley fever on a cattle farm in Bela-Bela, South Africa, in 2008

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    Lourenço P. Mapaco

    2012-04-01

    Full Text Available In 2008, a suspected outbreak of Rift Valley fever (RVF was reported on a farm in the Bela-Bela area, Limpopo Province, South Africa. Seven calves died on the affected dairy farm, where no RVF vaccination programme was practised. No apparent clinical disease was reported in the other 300 cattle (33 calves included or 200 sheep on the farm. During the outbreak, blood samples from 77.7% (233/300 of the cattle and 36.5% (73/200 of the sheep were collected on the affected farm and 55 blood samples were taken from cattle on a neighbouring farm. Eight weeks later, 78% of the cattle (234/300 and 42.5% of the sheep (85/200 were bled on the affected farm only. All sera were tested by an Immunoglobulin M (IgM-capture Enzymelinked immunosorbent assay (ELISA and by an indirect Immunoglobulin G (IgG ELISA. Selected IgM-positive (n = 14, IgG-positive (n = 23 and samples negative for both IgM and IgG-specific antibodies against RVF virus (n = 19 were tested using the serum neutralisation test (SNT. Sera from IgM-positive (n = 14 and negative (n = 20 animals were also tested by a TaqMan polymerase chain reaction (PCR. On the affected farm, 7% (16/233 of the cattle were IgM-positive and 13.7% (32/233 IgG-positive at the first bleed and 2% were IgM-positive at the second bleed, whilst the number of cattle positive for IgG-specific antibodies increased by 21.3% compared with the first bleed. Only 1.4% of sheep were positive for both IgM and IgG antibodies at the first collection; at the second bleed, IgM-positive cases decreased to 1.2%, whilst IgG-positive cases increased to 2.4%. Whilst no IgM-positive cattle were found on the neighbouring farm, 5.5% of cattle were IgG-positive. The SNT confirmed most of the ELISA results, whilst PCR results were all negative. Although serology results indicated virus circulation on both farms, the negative PCR results demonstrated that the animals were not viraemic at the time they were sampled. The movement of infected

  1. Significance of Neospora caninum in cattle farming

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    Ilić Tamara

    2006-01-01

    Full Text Available Neospora caninum is an obligate intracellular protozoan parasite which primarily causes diseases in dogs and cattle all over the world. It was first described in Norway in the mid-eighties in dogs, after which, until the present time, clinical neosporosis was proven in sheep, goats, deer, rhinoceroses, horses, and experimental rodents. Antibodies against N. caninum have been found also in the serum of water buffalo, red and gray foxes, coyotes, camels, and felines. Due to the similarity of this Coccidia with Toxoplasma gondi, the neosporosis was for a series of years incorrectly diagnozed as toxoplasmosis. Domestic canines, dogs, are the only real host for N. caninum. Its life cycle covers three stages of development: tachyzoites, tissue cysts and oocysts. Carnivores are infected by ingesting parts of infected tissue which contain tissue cysts with bradyzoites. The dominant pathway of transmission of this cause in cattle is transplacentary infection, but cattle can also be infected by ingestion of feed or water contaminated by sporulated oo-cysts of N. caninum. Bitches can be subclinical carriers of the parasite, when they pass on the cause transplacentarily, which results in more than one litter being born with the infection. Neosporosis today appears as the main cause of abortions and neonatal deaths in dairy cows and fattening cattle in almost all parts of the world, but with the highest incidence in the United States (US, New Zealand, The Netherlands, and Germany. The treatment of this disease has not been fully determined, but medicines used for the treatment of toxoplasmosis have yielded certain good results. There is no verified vaccine that would prevent undesired abortions in cattle. .

  2. Sensitive and selective spectrophotometric assay of piroxicam in pure form, capsule and human blood serum samples via ion-pair complex formation

    Science.gov (United States)

    Alizadeh, Nina; Keyhanian, Fereshteh

    2014-09-01

    A simple, accurate and highly sensitive spectrophotometric method has been developed for the rapid determination of piroxicam (PX) in pure and pharmaceutical formulations. The proposed method involves formation of stable yellow colored ion-pair complexes of the amino derivative (basic nitrogen) of PX with three sulphonphthalein acid dyes namely; bromocresol green (BCG), bromothymol blue (BTB), bromophenol blue (BPB) in acidic medium. The colored species exhibited absorption maxima at 438, 429 and 432 nm with molar absorptivity values of 9.400 × 103, 1.218 × 103 and 1.02 × 104 L mol-1 cm-1 for PX-BCG, PX-BTB and PX-BPB complexes, respectively. The effect of optimum conditions via acidity, reagent concentration, time and solvent were studied. The reactions were extremely rapid at room temperature and the absorbance values remained constant for 48 h. Beer’s law was obeyed with a good correlation coefficient in the concentration ranges 1-100 μg mL-1 for BCG, BTB complexes and 1-95 μg mL-1 for BPB complex. The composition ratio of the ion-pair complexes were found to be 1:1 in all cases as established by Job’s method. No interference was observed from common additives and excipients which may be present in the pharmaceutical preparations. The proposed method was successfully applied for the determination of PX in capsule and human blood serum samples with good accuracy and precision.

  3. Implementation of a SPR immunosensor for the simultaneous detection of the 22K and 20K hGH isoforms in human serum samples.

    Science.gov (United States)

    de Juan-Franco, Elena; Rodríguez-Frade, J M; Mellado, M; Lechuga, Laura M

    2013-09-30

    We have implemented a Surface Plasmon Resonance (SPR) immunosensor based on a sandwich assay for the simultaneous detection of the two main hGH isoforms, of 22 kDa (22K) and 20 kDa (20K). An oriented-antibody sensor surface specific for both hormone isoforms was assembled by using the biotin-streptavidin system. The immunosensor functionality was checked for the direct detection of the 22K hGH isoform in buffer, which gave high specificity and reproducibility (intra and inter-assay mean coefficients of variation of 8.23% and 9% respectively). The selective determination of the 22K and 20K hGH isoforms in human serum samples in a single assay was possible by using two specific anti-hGH monoclonal antibodies. The detection limit for both hormone isoforms was 0.9 ng mL(-1) and the mean coefficient of variation was below 7.2%. The excellent reproducibility and sensitivity obtained indicate the high performance of this immunosensor for implementing an anti-doping test. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Evaluation of antibodies against glycoprotein D (gD) and glycoprotein G (gG) in HSV-1 infected individuals' serum samples.

    Science.gov (United States)

    Meshkat, Z; Roostaee, M H; Soleimanjahi, M; Zandi, K

    2012-04-01

    Glycoproteins D (gD) and G (gG) of herpes simplex virus type 1 (HSV-1) are virus envelope glycoproteins that are able to induce HSV-1 antibody production in infected persons. Therefore, those proteins could be in interest to develop the serodiagnostic test(s) for HSV antibody detection. The aim of present study was the comparison of anti-gD and anti-gG antibodies in HSV-1 infected individuals' serum samples. In this study, recombinant gD and gG were prepared and used for western blot test to detect the antibodies against HSV-1. Our data showed the total gD antibody titer was higher than gG antibody titer in the HSV-1 infected patient's sera but the gG antibody titer was high significantly. According to our results, gD and gG can be used for designing the diagnostic laboratory tests to evaluate total antibody against HSV-1 and HSV-2.

  5. Bayesian Estimation of the True Prevalence and of the Diagnostic Test Sensitivity and Specificity of Enteropathogenic Yersinia in Finnish Pig Serum Samples

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    M. J. Vilar

    2015-01-01

    Full Text Available Bayesian analysis was used to estimate the pig’s and herd’s true prevalence of enteropathogenic Yersinia in serum samples collected from Finnish pig farms. The sensitivity and specificity of the diagnostic test were also estimated for the commercially available ELISA which is used for antibody detection against enteropathogenic Yersinia. The Bayesian analysis was performed in two steps; the first step estimated the prior true prevalence of enteropathogenic Yersinia with data obtained from a systematic review of the literature. In the second step, data of the apparent prevalence (cross-sectional study data, prior true prevalence (first step, and estimated sensitivity and specificity of the diagnostic methods were used for building the Bayesian model. The true prevalence of Yersinia in slaughter-age pigs was 67.5% (95% PI 63.2–70.9. The true prevalence of Yersinia in sows was 74.0% (95% PI 57.3–82.4. The estimates of sensitivity and specificity values of the ELISA were 79.5% and 96.9%.

  6. Neuropathology of organophosphate poisoning in dairy cattle

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    Yulvian Sani

    2007-03-01

    Full Text Available The purpose of this study is to investigate pathological changes in brain tissues of Frisien Holstein dairy cattle affected by organophosphate (OP. The study was directed to anticipate spongiform encephalopathy. Samples consisted of animal feeds, sera and brain tissues were collected from Lembang, West Java. Animal feeds (fodders and commercial feed were collected directly from the dairy farms around Lembang. Sera (31 samples were from dairy cattle owned by the local farmers and brain tissues were from the local animal slaughter house. Pesticide residues were analysed following a standard procedure using gas chromatography (GC. There was an interaction between pesticide residues in animal feeds, residue level of pesticides in sera and brain tissues to cause encephalopathy in dairy cattle. Pesticide contamination in animal feeds was regarded as the source of encephalopathy in dairy cattle. The total average of OP residues (16.8 ppb were lower than organochlorines/OC (18.7 ppb in fodder, showing that pesticides were originated from the contaminated soils. On the other hand, the total average of OP residues in commercial feeds (12.0 ppb, sera (85.6 ppb and brain tissues (22.7 ppb were higher than OC (1.8; 16.7; and 5.1 ppb. The OP appears more frequently used for dairy farm activity as insecticides. Histopathological examination for brain tissues of dairy cattle showed that most cattle were diagnosed as encephalopathy with microscopic changes of vacuolation, neuronal necrosis, chromatolysis of neurons and nucleolysis of neurons. The encephalopathy was confirmed in rats intoxicated with chlorpyrifos methyl as severe brain damage with spongiform-like lesions.

  7. Epidemiological profile of reproductive loss in dairy cattle

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    Raul Costa Mascarenhas Santana

    2012-12-01

    Full Text Available Several agents can be present in dairy cattle with a history of abortion as Neospora caninum, Leptospira spp, Bovine Herpesvirus Type 1 (BHV-1 and Brucella abortus. Some of these are considered transmitters cosmopolitan zoonosis of great economic impact and risk to human and animal health. The aim of this work was draw an epidemiological profile of reproductive losses and to determine the prevalence of antibodies against the main agents of reproductive diseases in dairy cattle. The study was conducted on a property in São Carlos city. For determination of reproductive failure, pre-existing data of abortion and stillbirths were analyzed from January 2006 to December 2011 on an average of 274 dairy cows of Holstein and crossbred Holstein-Jersey. On March 1, 2012 blood serum samples were collected of 142 breeding animals of ages above two years, in which 21.1% showed cases of abortions or stillbirths of at least one pregnancy. We used serologic tests of microscopic agglutination test, immunofluorescent antibody technique, serum neutralization technique, tamponated acidified antigen test for detection of anti-Leptospira spp and anti-Neospora caninum, anti-Bovine Herpesvirus Type-1 (BHV-1 and anti- Brucella abortus, respectively. The tests were performed at Universidade Estadual Paulista, Botucatu and Jaboticabal campi and EMBRAPA Southeast Livestock. During the study period, it was observed an average monthly rate of 1.7 abortions and 0.7 stillbirths, with an incidence of 63.6% and 58.0% of the cases observed, respectively, between November and April, period of higher pluviometric precipitation in the region. Among the cases of abortions observed, 76.2% happened between the fourth and sixth month of pregnancy. The serological tests carried out showed that 15.5% of the animals had titers greater than or equal to 1:200 of anti-Neospora caninum. Among the animals with a history of abortions or stillbirths, 28.58% and 11.22%, respectively, were serum

  8. Coxiella burnetii seroprevalence and associated risk factors in dairy and mixed cattle farms from Ecuador.

    Science.gov (United States)

    Carbonero, Alfonso; Guzmán, Lucía T; Montaño, Karen; Torralbo, Alicia; Arenas-Montes, Antonio; Saa, Luis R

    2015-03-01

    Q fever is a zoonotic disease caused by Coxiella burnetii, a bacterial agent for which ruminants are the main reservoir. An extensive cross-sectional study to determine the seroprevalence of and associated risk factors for Q fever was performed in dairy and mixed (dairy-beef) cattle herds in Ecuador. A total of 2668 serum samples from 386 herds were analyzed using an ELISA. In addition, a questionnaire with 57 variables related to management, feeding, facilities, biosecurity and animal health was completed for every cattle farm. A Generalized Estimating Equations model was used to determine the factors associated with C. burnetii seropositivity. The true prevalence of C. burnetii seropositivity in dairy and mixed cattle from Ecuador reached 12.6% (CI95%: 11.3-13.9%). The herd prevalence was 46.9% (181/386) (CI95%: 41.9-51.9%), and the within herd prevalence ranged between 8% and 100% (mean: 25.0%; Q1: 12.5%, Q2: 25.0%, Q3: 37.5%). Four factors were included in the GEE model for C. burnetii seropositivity: age of the cattle (OR: 1.01; CI95%: 1.006-1.014), feeding of calves with milk replacers (OR: 1.94; CI95%: 1.1-3.3), bovine respiratory syncytial virus seropositivity (OR: 1.54; CI95%: 1.1-2.3), and disinfection of the umbilical cord (OR: 0.60; CI95%: 0.4-0.9). Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Novel Secreted Antigens of Mycobacterium paratuberculosis as Serodiagnostic Biomarkers for Johne's Disease in Cattle

    Science.gov (United States)

    Facciuolo, Antonio; Kelton, David F.

    2013-01-01

    Johne's disease is a chronic gastroenteritis of cattle caused by Mycobacterium avium subsp. paratuberculosis that afflicts 40% of dairy herds worldwide. M. avium subsp. paratuberculosis-infected cattle can remain asymptomatic for years while transmitting the pathogen via fecal contamination and milk. Current serodiagnosis with enzyme-linked immunosorbent assays (ELISAs) fails to detect asymptomatic M. avium subsp. paratuberculosis-infected cattle due to the use of poorly defined antigens and knowledge gaps in our understanding of M. avium subsp. paratuberculosis components eliciting pathogen-specific immune responses. We set out to (i) define a subset of proteins that contain putative antigenic targets and (ii) screen these antigen pools for immunogens relevant in detecting infection. To accomplish our first objective, we captured and resolved M. avium subsp. paratuberculosis-secreted proteins using a 2-step fractionation method and reverse-phase liquid chromatography to identify 162 unique proteins, of which 66 had not been previously observed in M. avium subsp. paratuberculosis culture filtrates. Subsequent screening of M. avium subsp. paratuberculosis-secreted proteins showed four antigens, of which one or more reacted on immunoblotting with individual serum samples from 35 M. avium subsp. paratuberculosis-infected cows. Moreover, these novel antigens reacted with sera from 6 low M. avium subsp. paratuberculosis shedders and 3 fecal-culture-positive cows labeled as ELISA seronegative. The specificity of these antigens was demonstrated using negative-control sera from uninfected calves (n = 5) and uninfected cows (n = 5), which did not react to any of these antigens in immunoblotting. As three of the four antigens are novel, their characterization and incorporation into an ELISA-based format will aid in detecting asymptomatic cattle in early or subclinical stages of disease. PMID:24089453

  10. Conservation of the genetic material of Macedonian Busha cattle

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    Bunevski Gjoko

    2016-01-01

    Full Text Available The Busha is an indigenous breed of cattle in many Balkan countries. It has been bred for centuries. It belongs to primitive shorthorn cattle (Bos brachyceros europaeus. These cattle used to be the dominant and most important breed in almost all Balkan countries until the 1950s and 1960s, but today in lowland areas where intensive farming is practiced they have already been replaced by more productive and specialized breeds of cattle. In Macedonia this breed has officially been classified as a triple purpose breed (raised for meat, milk and draft but considering its low production capabilities it is more similar to some primitive draft breeds. This breed is part of the National Biodiversity Program for the conservation of indigenous breeds of animals in the Republic of Macedonia. Economic, cultural and scientific reasons underlie the need to protect the biological diversity of autochthonous breeds of cattle such as the Busha. The aim of the research was to establish a gene bank for different strains of adult Busha cattle in the Republic of Macedonia. To this end, 998 samples of blood, 1100 hair coat samples and 958 doses of semen were collected from adult Busha cattle. Also, a phenotypic characterization was done on adult Busha cattle for their major productive and morphological traits. During the last few years, there have been certain negative trends in the population size of Busha cattle in accordance with the decline of the rural population in the hills and uplands and young people's disinterest in rearing indigenous breeds of cattle such as the Busha.

  11. Epidemiology and genetic characterization of BVDV, BHV-1, BHV-4, BHV-5 and Brucella spp. infections in cattle in Turkey.

    Science.gov (United States)

    Aslan, Muhammet Eren; Azkur, Ahmet Kursat; Gazyagci, Serkal

    2015-11-01

    The aim of the study was to determine the epidemiological data of bovine viral diarrhea virus (BVDV), bovine herpesvirus-1 (BHV-1), bovine herpesvirus-4 (BHV-4), bovine herpesvirus-5 (BHV-5) and Brucella-associated cattle that were previously reported to have abortion and infertility problems in Ankara, Corum, Kirikkale and Yozgat provinces, Turkey. Whole blood and sera samples were obtained from 656 cattle, and antibodies against Brucella spp. were detected in 45 (6.86%) and 41 (6.25%) animals by Rose Bengal plate and serum tube agglutination tests, respectively. The seropositivity rates against BVDV, BHV-1 and BHV-4 were 70.89%, 41.3% and 28.78%, respectively. RT-PCR and PCR were performed to detect RNA and DNA viruses in blood samples, respectively. The BVDV 5'-untranslated region and BHV-1 gB gene detected in this study were phylogenetically analyzed. The BVDV strains analyzed in this study were closely related to those previously reported from Turkey. The nucleotide sequence from the BHV-1 strain detected in this study is the first nucleotide sequence of BHV-1 circulating in this area of Turkey deposited in the GenBank. The presence of Brucella spp. and prevalence of BHV-1, BHV-4 and BVDV in cattle should be further investigated throughout these regions.

  12. ELISA-based serological survey of Mycoplasma bovis in cattle in three local government areas in Adamawa State, Nigeria

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    Markus Isa Francis

    2015-06-01

    Full Text Available A serological survey for the detection of antibodies to Mycoplasma bovis was conducted in Adamawa State, Nigeria during the year 2012. A total of 400 serum samples were collected from three local government areas (LGA namely Yola (n=140, Mubi (n=130 and Ganye (n=130, and the samples were examined for the presence of M. bovis antibodies using BIO-X M. bovis antibody ELISA Kit. The overall seroprevalence of M. bovis was recorded as 19.5% (n=78/400. The highest seroprevalence was recorded in Ganye LGA (27.7%; n=36/130, followed by Yola LGA (20.0%; n=28/140 and Mubi LGA (10.8%; n=14/130. The cattle aging <1-year had the highest prevalence (25.8%, followed by 4-year-old cattle (22.4%. Based on sex distribution, 20.4% of the cows and 17.4% of the bulls were seropositive to M. bovis. Breed susceptibility to M. bovis infection showed that White Fulani had the highest prevalence (21.8%, and Sokoto Gudali had the lowest (11.9%. No statistical significant association was found between M. bovis infection and age, sex and breed of the cattle. In conclusion, the above findings are indicative for the presence of M. bovis in the study area.

  13. Anthelmintics for cattle.

    Science.gov (United States)

    Prichard, R K

    1986-07-01

    A number of anthelmintics are available for the control of gastrointestinal nematodes in cattle. In North America, O. ostertagi, Cooperia spp., lung worm, and F. hepatica probably cause the greatest losses in production. The older anthelmintics are often deficient in their action against some of these parasites. Recently, the Paratect morantel tartrate slow-release bolus has provided a mechanism for the prevention of infections with gastrointestinal nematodes and lung worm, to some extent, and this has been shown to produce considerable economic benefits. Fenbendazole removes arrested O. ostertagi larvae; thus, its availability is an important step in the prevention of type-2 ostertagiasis. It also has a very broad spectrum of activity that includes most other nematodes and tapeworms and is a very safe anthelmintic. Ivermectin is highly effective against almost all cattle nematodes and also has great value for the control of arthropod ectoparasites. In addition, it and levamisole are the only anti-nematode drugs that can be administered to cattle by injection. Clorsulon is a new, safe anthelmintic that provides good control of liver fluke and, thus, fills a gap in the control of helminths of cattle in North America. The efficient use of anthelmintics in association with management based on a knowledge of parasite epidemiology can ensure that cattle do not rapidly become re-infected. In this way, the benefits from the use of anthelmintics can be very considerable and far greater than the costs of control.

  14. Survey of serum concentrations of dioxins, furans, and coplanar polychlorinated biphenyls in a small non-random sample of U.S. residents

    Energy Technology Data Exchange (ETDEWEB)

    Grassman, J. [Brooklyn Coll. CUNY, Health and Nutrition Sciences, Brooklyn, New York, NY (United States); Patterson, D.G. Jr.; Needham, L.L. [National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA (United States); Spencer, D.L.; Masten, S.A. [Environmental Toxicology Program, National Inst. of Environmental Health Sciences, Research Triangle Park, NC (United States)

    2004-09-15

    This cross sectional assessment of serum dioxin concentrations was conducted as part of a larger study to examine the relationship between dioxin exposure and gene expression in peripheral blood mononuclear cells. Recent reports indicate that environmental levels of dioxins have declined since the mid-1980's. Except for the recent National Health and Nutrition Examination Survey (NHANES), there has been little systematic surveillance of serum dioxins levels in the US general population. Here, we report the serum concentrations of 22 congeners of dioxins and dioxin-like compounds and their relationship with age, sex, smoking, and meat consumption.

  15. Does the time interval between antimüllerian hormone serum sampling and initiation of ovarian stimulation affect its predictive ability in in vitro fertilization-intracytoplasmic sperm injection cycles with a gonadotropin-releasing hormone antagonist?

    DEFF Research Database (Denmark)

    Polyzos, Nikolaos P; Nelson, Scott M; Stoop, Dominic

    2013-01-01

    To investigate whether the time interval between serum antimüllerian hormone (AMH) sampling and initiation of ovarian stimulation for in vitro fertilization-intracytoplasmic sperm injection (IVF-ICSI) may affect the predictive ability of the marker for low and excessive ovarian response....

  16. Rapid detection of cytomegalovirus in bronchoalveolar lavage fluid and serum samples by polymerase chain reaction: correlation of virus isolation and clinical outcome for patients with human immunodeficiency virus infection

    DEFF Research Database (Denmark)

    Hansen, K K; Vestbo, Jørgen; Benfield, T

    1997-01-01

    Bronchoalveolar lavage (BAL) fluids and serum samples from 153 patients with pulmonary symptoms who were infected with human immunodeficiency virus (HIV) and underwent BAL were examined for the presence of cytomegalovirus (CMV) by conventional culture and by polymerase chain reaction (PCR...

  17. Genomic dairy cattle breeding

    DEFF Research Database (Denmark)

    Mark, Thomas; Sandøe, Peter

    2010-01-01

    The aim of this paper is to discuss the potential consequences of modern dairy cattle breeding for the welfare of dairy cows. The paper focuses on so-called genomic selection, which deploys thousands of genetic markers to estimate breeding values. The discussion should help to structure...... the thoughts of breeders and other stakeholders on how to best make use of genomic breeding in the future. Intensive breeding has played a major role in securing dramatic increases in milk yield since the Second World War. Until recently, the main focus in dairy cattle breeding was on production traits......, unfavourable genetic trends for metabolic, reproductive, claw and leg diseases indicate that these attempts have been insufficient. Today, novel genome-wide sequencing techniques are revolutionising dairy cattle breeding; these enable genetic changes to occur at least twice as rapidly as previously. While...

  18. Viral Communities Among Sympatric Vampire Bats and Cattle.

    Science.gov (United States)

    Escalera-Zamudio, Marina; Taboada, Blanca; Rojas-Anaya, Edith; Löber, Ulrike; Loza-Rubio, Elizabeth; Arias, Carlos F; Greenwood, Alex D

    2017-11-21

    Vampire bats are the only mammals known to feed exclusively on blood from other animals, often from domestic cattle. We tested the hypothesis that the adaptation of vampire bats to hematophagy would have resulted in shared viral communities among vampire bats and cattle, as a direct result of historic spillover events occurring due to hematophagy. We analyzed the presence of different viruses in sample populations of sympatric bat and prey populations and searched for shared viruses between taxa. A limited number of DNA viral groups were detected within each species. However, there was no evidence for a shared viral community among the vampire bat and cattle populations tested.

  19. Evaluation of the use of nonesterified fatty acids and β-hydroxybutyrate concentrations in pooled serum samples for herd-based detection of subclinical ketosis in dairy cows during the first week after parturition.

    Science.gov (United States)

    Borchardt, Stefan; Staufenbiel, Rudolf

    2012-04-15

    To evaluate the use of nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHBA) concentrations in pooled serum samples for herd-based detection of subclinical ketosis (SCK) in dairy cows after calving. Cross-sectional study. 1,100 dairy cows from 110 herds. Blood samples were collected from 10 healthy cows/herd in the first week after parturition. Aliquots of serum were mixed to create a pooled sample. Concentrations of NEFA and BHBA were measured to estimate prevalence of SCK. Pooled sample test results were compared with those obtained for individual samples. Linear regression and receiver-operating characteristic curve analysis were performed; Bland-Altman plots were used to evaluate agreement between methods. Overall prevalence of SCK was 30.7%, 19.3%, and 13.6%, as determined by use of BHBA threshold concentrations of 1,000, 1,200, and 1,400 μmol/L, respectively. Pooled sample concentrations of NEFA and BHBA were significantly correlated (r = 0.98 and 0.97, respectively) with individual sample means and with the number of cows that had NEFA (R(2) range, 0.81 to 0.84) or BHBA (R(2) range, 0.65 to 0.76) concentrations above predefined thresholds. Pooled sample concentrations of NEFA and BHBA were very accurate to highly accurate for herd-based detection of SCK. Analysis of NEFA and BHBA concentrations in pooled serum samples was useful for herd-based detection of SCK. A sample size of 10 cows/herd was deemed adequate for monitoring dairy herds for SCK. Reference criteria specific to pooled samples should be used for this type of herd-based testing.

  20. Development and validation of a rapid turboflow LC-MS/MS method for the quantification of LSD and 2-oxo-3-hydroxy LSD in serum and urine samples of emergency toxicological cases.

    Science.gov (United States)

    Dolder, Patrick C; Liechti, Matthias E; Rentsch, Katharina M

    2015-02-01

    Lysergic acid diethylamide (LSD) is a widely used recreational drug. The aim of the present study is to develop a quantitative turboflow LC-MS/MS method that can be used for rapid quantification of LSD and its main metabolite 2-oxo-3-hydroxy LSD (O-H-LSD) in serum and urine in emergency toxicological cases without time-consuming extraction steps. The method was developed on an ion-trap LC-MS/MS instrument coupled to a turbulent-flow extraction system. The validation data showed no significant matrix effects and no ion suppression has been observed in serum and urine. Mean intraday accuracy and precision for LSD were 101 and 6.84%, in urine samples and 97.40 and 5.89% in serum, respectively. For O-H-LSD, the respective values were 97.50 and 4.99% in urine and 107 and 4.70% in serum. Mean interday accuracy and precision for LSD were 100 and 8.26% in urine and 101 and 6.56% in serum, respectively. For O-H-LSD, the respective values were 101 and 8.11% in urine and 99.8 and 8.35% in serum, respectively. The lower limit of quantification for LSD was determined to be 0.1 ng/ml. LSD concentrations in serum were expected to be up to 8 ng/ml. 2-Oxo-3-hydroxy LSD concentrations in urine up to 250 ng/ml. The new method was accurate and precise in the range of expected serum and urine concentrations in patients with a suspected LSD intoxication. Until now, the method has been applied in five cases with suspected LSD intoxication where the intake of the drug has been verified four times with LSD concentrations in serum in the range of 1.80-14.70 ng/ml and once with a LSD concentration of 1.25 ng/ml in urine. In serum of two patients, the O-H-LSD concentration was determined to be 0.99 and 0.45 ng/ml. In the urine of a third patient, the O-H-LSD concentration was 9.70 ng/ml.

  1. Fasting Serum Glucose and Cholesterol as Predictors of Cardiovascular Reactivity to Acute Stress in a Sample of African American College Students.

    Science.gov (United States)

    Clark, Vernessa R; Perkins, Patrice; Carson, Bernice L; Boyd, Kimberly; Jefferson, Trayce M

    2015-01-01

    African Americans are at a greater risk of cardiovascular hyperactivity to stress than Caucasians; however the risk factors for this activity are not clearly delineated for African Americans. The purpose of this study was to determine the ability of fasting serum cholesterol concentration and fasting serum glucose (FSG) to predict cardiovascular reactivity to stress in African Americans. Serum cholesterol concentration and FSG levels were measured in 48 (40 women, 8 men) African American college students aged 18-30 years. Heart rate, cardiac output, stroke volume, mean arterial pressure and systolic and diastolic blood pressure were measured as the participants viewed a racially noxious scene on a digital video disc. Measurements were taken prior to the scene (pre stressor period), during the scene (stressor period), and while the participant recovered from the scene (recovery period). A multiple regression analysis revealed that total serum cholesterol and LDL significantly predicted diastolic blood pressure during the pre-stressor period. FSG significantly predicted mean arterial pressure during the recovery period, and predicted stroke volume during the pre-stressor period, stressor period, and the recovery period. FSG was a better predictor of cardiovascular reactivity to stress than serum cholesterol concentration, predicting mean arterial pressure and stroke volume. This finding may be due to the association of glucose with diabetes, which is more prevalent in African Americans.

  2. Analytical Bias Exceeding Desirable Quality Goal in 4 out of 5 Common Immunoassays: Results of a Native Single Serum Sample External Quality Assessment Program for Cobalamin, Folate, Ferritin, Thyroid-Stimulating Hormone, and Free T4 Analyses.

    Science.gov (United States)

    Kristensen, Gunn B B; Rustad, Pål; Berg, Jens P; Aakre, Kristin M

    2016-09-01

    We undertook this study to evaluate method differences for 5 components analyzed by immunoassays, to explore whether the use of method-dependent reference intervals may compensate for method differences, and to investigate commutability of external quality assessment (EQA) materials. Twenty fresh native single serum samples, a fresh native serum pool, Nordic Federation of Clinical Chemistry Reference Serum X (serum X) (serum pool), and 2 EQA materials were sent to 38 laboratories for measurement of cobalamin, folate, ferritin, free T4, and thyroid-stimulating hormone (TSH) by 5 different measurement procedures [Roche Cobas (n = 15), Roche Modular (n = 4), Abbott Architect (n = 8), Beckman Coulter Unicel (n = 2), and Siemens ADVIA Centaur (n = 9)]. The target value for each component was calculated based on the mean of method means or measured by a reference measurement procedure (free T4). Quality specifications were based on biological variation. Local reference intervals were reported from all laboratories. Method differences that exceeded acceptable bias were found for all components except folate. Free T4 differences from the uncommonly used reference measurement procedure were large. Reference intervals differed between measurement procedures but also within 1 measurement procedure. The serum X material was commutable for all components and measurement procedures, whereas the EQA materials were noncommutable in 13 of 50 occasions (5 components, 5 methods, 2 EQA materials). The bias between the measurement procedures was unacceptably large in 4/5 tested components. Traceability to reference materials as claimed by the manufacturers did not lead to acceptable harmonization. Adjustment of reference intervals in accordance with method differences and use of commutable EQA samples are not implemented commonly. © 2016 American Association for Clinical Chemistry.

  3. Risk factors associated with increased bovine leukemia virus proviral load in infected cattle in Japan from 2012 to 2014.

    Science.gov (United States)

    Ohno, Ayumu; Takeshima, Shin-nosuke; Matsumoto, Yuki; Aida, Yoko

    2015-12-02

    Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis, a malignant B cell lymphoma. BLV has spread worldwide and causes serious problems. After infection, the BLV genome is integrated into the host DNA and can be amplified during periods of latency. We previously designed degenerate primers using the Coordination of Common Motifs (CoCoMo) algorithm to establish a new quantitative real-time PCR method (BLV-CoCoMo-qPCR-2) of measuring the proviral load of both known and novel BLV variants. Here, we aimed to examine the correlation between proviral load and risk factors for BLV infection, such as breeding systems, parousity, and colostrum feeding. Blood and serum samples were collected from 83 BLV-positive farms in 22 prefectures of Japan, and the BLV proviral load and anti-BLV antibody levels were measured. BLV was detected in 73.3% (1039/1,417) of cattle by BLV-CoCoMo-qPCR-2 and the provirus was detected in 93 of 1039 antibody-negative samples. The results showed that the proviral load increased with progression of lymphocytosis. Next, the risk factors associated with increasing BLV infection rate were examined along with any association with proviral load. The proviral load was higher in cattle with lymphocytosis than in healthy cattle, and higher in multiparous cows than in nulliparous cows. Finally, proviral loads were higher in contact breeding systems than in non-contact breeding systems. Taken together, these findings may help to formulate a plan for eliminating BLV from contaminated farms. This is the first nationwide study to estimate BLV proviral load in Japanese cattle. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Clinical haematology and biochemistry profiles of cattle naturally infected with Theileria orientalis Ikeda type in New Zealand.

    Science.gov (United States)

    Lawrence, K E; Forsyth, S F; Vaatstra, B L; McFadden, Amj; Pulford, D J; Govindaraju, K; Pomroy, W E

    2018-01-01

    To present the haematology and biochemistry profiles for cattle in New Zealand naturally infected with Theileria orientalis Ikeda type and investigate if the results differed between adult dairy cattle and calves aged biochemistry results were obtained from blood samples from cattle which tested positive for T. orientalis Ikeda type by PCR, that were submitted to veterinary laboratories in New Zealand between October 2012 and November 2014. Data sets for haematology and biochemistry results were prepared for adult dairy cattle (n=62 and 28, respectively) and calves aged biochemistry (pbiochemistry changes observed in cattle infected with T. orientalis Ikeda type were consistent with extravascular haemolytic anaemia. Adult dairy cattle were more likely to be severely anaemic than calves. There were differences in haematology and biochemistry profiles between adult dairy cattle and calves, but most of these differences likely had a physiological rather than pathological basis. Overall, the haematological changes in calves aged <6 months appeared less severe than in adult dairy cattle.

  5. Prevalence of hemoprotozoan diseases in cattle population of chittagong division, Bangladesh

    DEFF Research Database (Denmark)

    Alim, Md. Abdul; Das, Shubhagata; Roy, Krisna

    2012-01-01

    A one year (2009-10) prevalence study on hemoprotozoan diseases was conducted in crossbred and indigenous cattle, Chittagong, Bangladesh. Blood samples were collected randomly from 216 crossbred and 432 indigenous cattle of four representative areas in three consecutive seasons. Samples were...

  6. Comparative Brucella abortus antibody prevalence in cattle under contrasting husbandry practices in Uganda

    Directory of Open Access Journals (Sweden)

    Gerald Nizeyimana

    2013-02-01

    Full Text Available A study was conducted in the Luwero and Nakasongola districts in central Uganda to determine and compare the prevalence and distribution of antibodies against Brucella abortus in cattle under contrasting husbandry practices, using two serological tests. Three hundred and fifteen serum samples were systematically sampled from 29 farms and subsequently tested using the Rose Bengal plate test (RBPT and Indirect Antibody Enzyme Linked Immunosorbent Assay (I-ELISA. The overall prevalence of antibodies against Brucella abortus in the Nakasongola and Luwero districts was 2.4% and 4.7% on RBPT, compared with 1.2% and 3.34 % on I-ELISA. There was no significant difference between the results obtained by RBPT and indirect antibody ELISA (p > 0.05. It was noted that antibodies against Brucella abortus were widely spread over different farms regardless of the cattle grazing system (p > 0.05. Based on the findings, it is feasible to use RBPT as a cheaper screening alternative for brucellosis. A comprehensive national brucellosis study should be undertaken to study the epidemiology and prevalence of brucellosis in Uganda.

  7. Bovine herpes virus-1 (BoHV-1 detection in dairy cattle with reproductive problems in Sudan

    Directory of Open Access Journals (Sweden)

    Amira Mohamed Elhassan

    2015-06-01

    Full Text Available The present work aimed to observe the infection pattern of Bovine herpes virus-1 (BoHV-1 in dairy cattle with reproductive problems in Sudan. A total of 140 samples comprising of vaginal swab (n=97, placenta (n=15, whole blood (n=19, uterine fluid (n=1, and serum (n=8 were collected from 16 dairy herds showing particularly high rate of abortion and infertility in Khartoum State. The samples were used for virus isolation, and were tested by Enzyme-Linked Immunosorbent Assay (ELISA and polymerase chain reaction (PCR. No virus could be isolated from the samples inoculated for isolation in cell culture. Out of 80 specimens tested by ELISA, 7 (8.75% were found to be positive, and one sample was doubtful. Using PCR, 11 (10.7% out of 103 samples were found to be positive. When comparing between two methods for DNA extraction, the DNA extracted by commercial kit was found to be better in quality as compared to the DNA extracted using phenol/chloroform/isoamyl-alcohol method. The study confirmed the presence of BoHV-1 in cattle farms with reproductive problems in Sudan.

  8. On-line coupling of solid-phase extraction with mass spectrometry for the analysis of biological samples III. Determination of prednisolone in serum

    NARCIS (Netherlands)

    van Hout, M.W.J.; Hofland, C.M; Niederlander, H.A G; Bruins, A.P.; de Zeeuw, R.A.; de Jong, G.J.

    2003-01-01

    Solid-phase extraction (SPE) was directly coupled to mass spectrometry (MS) to assess the feasibility of the system for the rapid determination of prednisolone in serum. A C-18 stationary phase allowed washing of the cartridge with 25% methanol. Elution was performed by switching the methanol

  9. Neutralizing antibodies against two HIV-1 strains in consecutively collected serum samples: cross neutralization and association to HIV-1 related disease

    DEFF Research Database (Denmark)

    Arendrup, M; Nielsen, C M; Hansen, J E

    1992-01-01

    97 sera collected during a 10-year period from 10 HIV-1 infected individuals were tested for neutralizing capacity against a virus isolate FICPH-22 obtained from a Danish AIDS patient, and the laboratory strain HTLV-IIIB. Three patterns of serum neutralizing activity were demonstrated: (a) patients...

  10. Sample preparation of human serum for the analysis of tumor markers : Comparison of different approaches for albumin and gamma-globulin depletion

    NARCIS (Netherlands)

    Govorukhina, N I; Keizer-Gunnink, A; van der Zee, A G J; de Jong, Steven; de Bruijn, H W A; Bischoff, R

    2003-01-01

    LC-MS is a powerful method for the sensitive detection of proteins and peptides in biological fluids. However, the presence of highly abundant proteins often masks those of lower abundance and thus generally prevents their detection and identification in proteomic studies. In human serum the most

  11. Glaucoma and vitamins A, C, and E supplement intake and serum levels in a population-based sample of the United States.

    Science.gov (United States)

    Wang, S Y; Singh, K; Lin, S C

    2013-04-01

    To investigate the potential association between glaucoma prevalence and supplemental intake, as well as serum levels of vitamins A, C and E. This cross-sectional study included 2912 participants in the 2005-2006 National Health and Nutrition Examination Survey, age ≥40 years, who self-reported a presence or absence of glaucoma. Participants were interviewed regarding the use of dietary supplements during the preceding 30-day period. Participants also underwent serum measurements of vitamins A, C, and E (both alpha- and gamma-tocopherol). Information on the primary outcome measure, presence or absence of glaucoma, as well as demographic information, comorbidities and health-related behaviors, was assessed via interview. Multivariate odds ratios for self-reported glaucoma, comparing the highest quartile of consumption to no consumption, and adjusted for potential confounding variables were 0.48 (95% confidence interval (CI) 0.13-1.82) for vitamin A, 0.47 (95% CI 0.23-0.97) for vitamin C, and 2.59 (95% CI 0.89-7.56) for vitamin E. Adjusted odds ratios for self-reported glaucoma comparing the highest vs lowest quintiles of vitamin serum levels were 1.44 (95% CI 0.79-2.62) for vitamin A, 0.94 (95% CI 0.42-2.11) for vitamin C, 1.40 (95% CI 0.70-2.81) for alpha-tocopherol, and 0.64 (95% CI 0.24-1.70) for gamma-tocopherol. Neither supplementary consumption with nor serum levels of vitamins A and E were found to be associated with glaucoma prevalence. While low- and high-dose supplementary consumption of vitamin C was found to be associated with decreased odds of glaucoma, serum levels of vitamin C did not correlate with glaucoma prevalence.

  12. Spontaneous coffee senna poisoning in cattle: report on 16 outbreaks

    Directory of Open Access Journals (Sweden)

    Priscila M.S. Carmo

    2011-02-01

    Full Text Available Sixteen outbreaks of Senna occidentalis (coffee senna that occurred in cattle in the state of Rio Grande do Sul, Brazil, were reviewed. The great majority (75% of the outbreaks occurred in adult cattle at pasture during the autumn and winter months with 50% in May, evidencing a striking seasonality. Mortality rates varied from 4.2% to 55.2% and cattle died 2 days up to 2 weeks after showing clinical signs that included dry feces (occasionally diarrhea, muscle weakness, reluctance to move, tachypnea, instability of the hind limbs with dragging of the toes, tremors in muscles of the thighs, neck, and head, ear dropping, sternal recumbency, lateral recumbency and death. Myoglobinuria characterized by a dark red or black discolored urine was a consistent finding in cattle affected at pasture but not in those poisoned by ration contaminated with coffee senna beans. Creatine phosphokinase serum activity was marked ly elevated. Main gross changes observed in 23 necropsies involved skeletal muscles of the hind limbs. These changes consisted of varying degrees of paleness of muscle groups. Subepicardial and subendocardial hemorrhages were present in the hearts of all affected cattle. Histologically a segmental degenerative myopathy of striated muscles was present in every case and had a multifocal polyphasic or monophasic character. Myocardial (3/23, hepatic (3/13, renal (3/10, and splenic (1/6 microscopic lesions were observed occasionally. Myocardial lesions were mild and consisted of vacuolation of cardiomyocytes or focal fibrosis. Hepatic changes consisted of diffuse hepatocelular vacuolation, cytosegrosomes within hepatocytes, and individual hepatocellular necrosis. Kidneys had vacuolar degeneration of tubular epithelium associated with acidophilic casts (proteinosis within tubular lumina. In the spleen there was marked necrosis of lymphocytes of the white pulp. No histological changes were found in the brains of 13 affected cattle. The data of this

  13. Perceptions of Farmers against Liquid Fertilizer Benefits of Beef Cattle Urine

    OpenAIRE

    Sirajuddin, Sitti Nurani; kasim, kasim; moh saleh, Ikrar

    2014-01-01

    The aim of this study was to know the perception of livestock farmers on the use of liquid organic fertilizer from urine of cattle at Sinjai Regency, South Sulawesi Province. The choice of location for a farmer group manufactures and markets liquid organic fertilizer from cattle urine. This research was conducted in May to July 2013.The population were all livestock farmers who use organic liquid fertilizer from cattle urine samples while livestock farmers who are directly involved in the ma...

  14. Molecular and Phylogenetic Analysis of Bovine Papillomavirus Type 1: First Report in Iraqi Cattle

    OpenAIRE

    Mohammed A Hamad; Al-Shammari, Ahmed M; Odisho, Shoni M.; Yaseen, Nahi Y.

    2016-01-01

    This study aimed to provide the first molecular characterization of bovine papillomavirus type 1 (BPV-1) in Iraq. BPV is a widely spread oncogenic virus in Iraqi cattle and is associated with the formation of both benign and malignant lesions, resulting in notable economic losses in dairy and beef cattle. In the current study, 140 cutaneous papilloma specimens were collected from cattle in central Iraq. These samples were submitted to histopathological examination, PCR, and sequencing analysi...

  15. Determination of Diagnostic Antigens in Cattle Amphistomiasis Using Western Blotting

    Directory of Open Access Journals (Sweden)

    A Halajian

    2009-05-01

    Full Text Available "nBackground: Mixed infection with amphistomes seems common in native cattle of Iran. The aim of this study was to determine diagnostic antigens in cattle mixed amphistomiasis."nMethods: Specific antigens of Cotylophoron cotylophorum, Gastrothylax crumenifer and Paramphisto­mum cervi (mixed infection, the most common species, were collected from cattle was deter­mined. Adult trematodes were collected from the rumen of naturally infected cattle at meat inspec­tion. After their homogenization and centrifugation, somatic antigens were prepared and ana­lyzed by SDS-PAGE. Specific antigens were determinated by western blot with homologous and heterolo­gous sera. SDS-PAGE of whole worms extract was performed at different concentrations and subse­quent gels staining. Immunoblotting analysis using sera from cattle naturally infected with am­phistomes, Dicrocoelium dendriticum, Fasciola spp. and hydatid cyst was performed."nResults: Electrophorese analysis of somatic antigens revealed the presence of 10 and 21 protein bands at 4 µgr/ml and 8 µgr/ml with molecular weights ranging from 25-120 and 25-150 kDa, respectively. The best result was taken at 8 mg/ml concentration. Although western blot of these proteins demon­strate 5 major antigenic polypeptides ranging from 50 to 100 kDa which were recognized by serum of cat­tle naturally infected with mixed amphistomes.

  16. The effects of bovine viral diarrhoea virus on cattle reproduction in relation to disease control.

    Science.gov (United States)

    Fray, M D; Paton, D J; Alenius, S

    2000-07-02

    Bovine viral diarrhoea virus (BVDV) is a major reproductive pathogen in cattle. Infection of the bull can lead to a fall in semen quality and the isolation of infectious virus in the ejaculate, while infection in the cow leads to poor conception rates, abortions and congenital defects. BVDV also reduces the animal's resistance to other respiratory and enteric pathogens. The prevalence of BVDV is primarily due to the efficiency with which the virus crosses the placenta of susceptible females. Calves that survive infection during the first trimester of pregnancy are born with a persistent and lifelong infection. These persistently infected (PI) animals represent between 1.0% and 2.0% of the cattle population and continuously shed infectious virus. The availability of reliable diagnostic ELISA and PCR techniques, which can test milk or serum samples for virus or antibodies, has simplified BVDV surveillance and improved the prospects for control. Although PI animals are the principal vectors within and between herds, they can be readily identified and removed. By contrast, cows carrying a PI foetus are particularly problematic. These animals have been compared to 'Trojan Horses' because they are virus-negative and antibody-positive but they deliver PI calves. In general, acutely infected cattle are much less efficient vectors but infections at the onset of puberty have resulted in a localised and persistent infection within the testes. Under these circumstances, virus shedding into the semen may remain undetected. Transmission of BVDV can be controlled through vaccination or eradication. BVDV vaccine technology has been developing over the past 30 years, but currently available vaccines are still of the conventional inactivated or attenuated sort. In general, vaccination has not been applied with sufficient rigor to make a significant impact on the level of circulating virus, unlike the national and regional eradication programmes established in areas such as

  17. Prevalence of cysticercosis in Estonian pigs and cattle

    DEFF Research Database (Denmark)

    Dorbek-Kilin, Elisabeth; Åhlberg, Tuuli; Tummeleht, Lea

    2018-01-01

    must be registered and reported when found. Our aim was to estimate the prevalence of cysticercosis in Estonia caused by T. solium in pigs and T. saginata in cattle. The four slaughterhouses participating in the study slaughter between them approximately 80% of pigs and cattle in Estonia annually....... Sampling spanned from February to April 2014, visiting the slaughterhouses five times per week. Visual inspection, palpation, and incisions at predilection sites were used to find cysts in both species. The sites inspected in both species were the external masseter, tongue, heart, and diaphragm....... In addition, the internal masseter in pigs was examined, and the internal pterygoid muscle and esophagus in cattle. DNA was extracted from the cysts and used for PCR amplification of the cox1-gene for Taenia genus and species identification. A total of 564 cattle and 1217 pigs were examined. Cysts were found...

  18. Evaluation of the hormones responsible for the gastrointestinal motility in cattle with displacement of the abomasum; ghrelin, motilin and gastrin.

    Science.gov (United States)

    Ozturk, A S; Guzel, M; Askar, T K; Aytekin, I

    2013-06-15

    This study provides the evidence of increased serum gastrointestinal motility hormone concentrations including ghrelin, motilin and gastrin in cattle with displacement of abomasum (DA). In this study, 38 cows with DA (21 left DA (LDA) and 17 right DA (RDA)) and 15 healthy controls were included. All cattle with DA were at the stage of postpartum one to eight weeks, and had clinical signs including anorexia, decreased milk yield and scanty, pasty faeces. Serum ghrelin, motilin and gastrin concentrations, and leptin concentration which is a functional antagonist of ghrelin, were determined by ELISA. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), Na, K, Cl, Ca and P concentrations were measured by spectrophotometer. In serum biochemical analysis, increases were seen on the serum ALT, AST and GGT activities; however, serum Na, K, Cl and P concentrations decreased in abomasal displacement compared with the control animals. The serum ghrelin, motilin and gastrin concentrations increased in the cattle with LDA and RDA, as compared with those in the healthy controls. On the other hand, serum leptin concentration decreased in the cattle with DA compared with the controls. Increases in the serum ghrelin, motilin and gastrin concentrations might be attributed to activation of gastrointestinal motility hormones to enhance of gastric emptying in impaired gastric motility and/or outlet occlusion in displaced abomasum.

  19. Husbandry risk factors associated with subclinical coccidiosis in young cattle.

    Science.gov (United States)

    Mitchell, E S E; Smith, R P; Ellis-Iversen, J

    2012-07-01

    This paper describes an observational longitudinal study of cattle farms in England and Wales, which aimed to identify management practices associated with the presence of Eimeria spp. infection in young cattle. Thirty cattle farms situated in England and Wales were selected and one group of more than 20 young cattle aged 5-18 months of age was monitored on each farm. Three variables were identified as significantly associated with status in a multivariable model. The odds of finding Eimeria spp. were lower on farms that kept sheep on the same premises as the cattle, as was an increase in the maximum age within the sampled group. The latter probably reflects the development of post-infection immunity within the sampled animals. Good water-trough hygiene protected against Eimeria spp. oocyst excretion, with the odds of detection being higher on farms where it was reported that the water troughs were not cleaned and emptied more than once per month. The value of frequent emptying and cleaning of water troughs in reducing the exposure of calves to Eimeria spp. and thus lowering the impact of coccidiosis, both clinical and subclinical should be communicated to cattle farmers. Crown Copyright © 2011. Published by Elsevier Ltd. All rights reserved.

  20. Serological survey of Neospora caninum infection in dairy cattle herds in Aguascalientes, Mexico.

    Science.gov (United States)

    García-Vázquez, Z; Cruz-Vázquez, C; Medina-Espinoza, L; García-Tapia, D; Chavarria-Martinez, B

    2002-06-03

    A serological survey for antibody activity to Neospora caninum was carried out in Aguascalientes, a state in the central part of Mexico. One-hundred and eighty-seven serum samples from 13 dairy herds were tested by the ELISA test. The herd prevalence was 100% and the overall prevalence was 59% (n=110). Seventy-six of 97 seropositive cows had previous records of abortion. There was a statistically significant difference between the groups (P<0.05). However, the odds ratio was 1.4, suggesting an association between abortion and seropositivity. Neosporosis in dairy cattle appears to be widespread in Mexico, warranting more epidemiological studies to determine the distribution of the causative protozoan.

  1. Citrus pulp for cattle.

    Science.gov (United States)

    Arthington, John D; Kunkle, William E; Martin, Amy M

    2002-07-01

    Citrus pulp is classified as an energy concentrate by-product feed. Citrus by-products fed to beef cattle include citrus molasses, citrus meal, wet citrus pulp, dried citrus pulp, and pelleted citrus pulp; however, in current production systems, pulp (wet, dry, and pelleted) is the only by-product commonly used. Citrus pulp production in the United States is limited to specific subtropical regions, of which south central Florida remains the largest with additional production in California and Texas.

  2. Salmonella in Swedish cattle

    OpenAIRE

    Ågren, Estelle

    2017-01-01

    In Sweden, all herds detected with salmonella are put under restrictions and measures aiming at eradication are required. The purpose of these studies was to provide a basis for decisions on how surveillance and control of salmonella in Swedish cattle can be made more cost-efficient. Results from a bulk milk screening were used to investigate seroprevalence of salmonella and to study associations between salmonella status and geographical location, local animal density, number of test pos...

  3. A survey of Western Australian sheep, cattle and kangaroos to determine the prevalence of Coxiella burnetii.

    Science.gov (United States)

    Banazis, Michael Janis; Bestall, Abbey Simone; Reid, Simon Andrew; Fenwick, Stan Gordon

    2010-07-14

    The objective of this study was to investigate the prevalence of Coxiella burnetii in two domestic ruminant species (cattle and sheep) and the western grey kangaroo (Macropus fuliginosus) in Western Australia (WA). The IDEXX CHEKiT Q Fever ELISA and CFT were used to test sera from 50 sheep and 329 head of cattle for anti-C. burnetii antibodies and 343 kangaroo sera were tested using an indirect ELISA developed specifically for this study. Faecal or urine samples collected from the same animals were tested with two PCR assays to identify active shedding of C. burnetii in excreta. Only two of the 379 ruminant sera had detectable levels of anti-C. burnetii antibodies according to the ELISA while the CFT did not detect any positive samples. In contrast 115 of the 343 western grey kangaroo serum samples were positive when tested with the antibody-ELISA. The first qPCR assay, targeting the IS1111a element, identified 41 of 379 ruminant and 42 of 343 kangaroo DNA samples as positive for C. burnetii DNA. The second qPCR, targeting the JB153-3 gene, identified nine C. burnetii DNA-positive ruminant samples and six positive kangaroo samples. Sequence comparisons showed high degrees of identity with C. burnetii. Isolation of C. burnetii from faeces was also attempted but was not successful. From the results presented here it appears that domestic ruminants may not be the most significant reservoir of C. burnetii in WA and that kangaroos may pose a significant threat for zoonotic transfer of this pathogen. (c) 2009 Elsevier B.V. All rights reserved.

  4. Evaluation of cardiac troponin I alterationsin dairy cattle with septicmetritis

    Directory of Open Access Journals (Sweden)

    majid fartashvand

    2013-11-01

    Full Text Available Metritis is an important disease in dairy cattle which causes economical loses including decrease in milk yield, increase calving interval, treatment costs and death of ill cases. Septic metritis usually occurs within 2-10 days after parturition, and characterized clinically with sever toxemia associated with purulent odorous uterine discharge with or without retained placenta. In this study, serum levels of cTnI were measured in 50 female Holstein cattle with septicmetritis and compared with normal cows. cTnI of serum in disease and control groups were 0.017 ± 0.008 and 0.005 ± 0.000 ng/dl, respectively. Heart rate, respiratory rate and rectal temperature in disease cases were significantly higher than normal cattle. There was significant correlation with cTnI and heart rate and rectal temperature. Endotoxemia is one of possible reasons of elevation of serum cTnI. Cytokines and endotoxins originated from gram negative bacteria that cause myocardium depression and ventricular dilatation. Furthermore impairment of left ventricle function is a significant effect of septic shock.

  5. Serosurvey of Crimean-Congo Hemorrhagic Fever Virus in Cattle, Mali, West Africa.

    Science.gov (United States)

    Maiga, Ousmane; Sas, Miriam Andrada; Rosenke, Kyle; Kamissoko, Badian; Mertens, Marc; Sogoba, Nafomon; Traore, Abdallah; Sangare, Modibo; Niang, Mamadou; Schwan, Tom G; Maiga, Hamidou Moussa; Traore, Sekou F; Feldmann, Heinz; Safronetz, David; Groschup, Martin H

    2017-06-01

    AbstractCrimean-Congo hemorrhagic fever is a tick-borne disease caused by the arbovirus Crimean-Congo hemorrhagic fever virus (CCHFV, family Bunyaviridae, genus Nairovirus ). CCHFV can cause a severe hemorrhagic fever with high-case fatality rates in humans. CCHFV has a wide geographic range and has been described in around 30 countries in the Middle East, Asia, Europe, and Africa including Mali and neighboring countries. To date, little is known about the prevalence rates of CCHFV in Mali. Here, using banked bovine serum samples from across the country, we describe the results of a seroepidemiological study for CCHFV aimed at identifying regions of circulation in Mali. In total, 1,074 serum samples were tested by a modified in-house CCHFV-IgG-enzyme-linked immunosorbent assay (ELISA) with confirmatory testing by commercial ELISA and immunofluorescence assay. Overall, 66% of samples tested were positive for CCHFV-specific IgG antibodies. Regional seroprevalence rates ranged from 15% to 95% and seemed to correlate with cattle density. Our results demonstrate that CCHFV prevalence is high in many regions in Mali and suggest that CCHFV surveillance should be established.

  6. Brucellosis in working equines of cattle farms from Minas Gerais State, Brazil.

    Science.gov (United States)

    Junqueira, Danilo Guedes; Dorneles, Elaine Maria Seles; Gonçalves, Vitor Salvador Picão; Santana, Jordana Almeida; Almeida, Valéria Maria de Andrade; Nicolino, Rafael Romero; Silva, Marcos Xavier; Mota, Ana Lourdes Arrais de Alencar; Veloso, Flávio Pereira; Stynen, Ana Paula Reinato; Heinemann, Marcos Bryan; Lage, Andrey Pereira

    2015-10-01

    The present survey aimed at estimating the seroprevalence of brucellosis in working equines of cattle farms from Minas Gerais State, Brazil, and investigating risk factors associated with the infection. Serum samples from 6439 animals, including 5292 horses, 1037 mules and 110 donkeys, were collected from 1936 herds, between September 2003 and March 2004, in 848 municipalities from the state of Minas Gerais, Brazil. The prevalence of antibodies against smooth Brucella spp. found in equines from Minas Gerais State was 1.37% (95% CI: 0.97-1.78), resulting in a prevalence of herds with infected animals of 4.28% (95% CI: 4.21-4.36). There were differences between regions but these were not of major epidemiological relevance nor were most of them statistically significant, given the considerable overlap of confidence intervals. Nevertheless, the point estimates suggest that the three northeastern regions have slightly higher prevalence than the rest of the state, both at the herd and animal levels. No association of Brucella spp. seropositivity with sex, age or host was observed. In conclusion, the present study showed a low but widespread prevalence of antibodies against smooth Brucella in equines kept in cattle farms in Minas Gerais, a state where bovine brucellosis is also widespread albeit with low prevalence. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Endocrine patterns in two strains of Japanese black cattle with growth retardation.

    Science.gov (United States)

    Takasu, Masaki; Hagiwara, Youko; Ohba, Yasunori; Nishii, Naohito; Hosoda, Iwai; Kitoh, Katsuya; Katoh, Kazuo; Kitagawa, Hitoshi

    2005-11-01

    Endocrine patterns were compared in 2 strains of Japanese black cattle with growth retardation; MHO- and HSK-paternal strains (MHO and HSK cattle, respectively). MHO cattle (n=8) displayed lower serum concentrations of insulin-like growth factor-1 (IGF-1), triiodothyronine (T3), thyroxine (T4), and cortisol (31.1+/-20.7 ng/ml, 73.9+/-51.9 ng/dl, and 2.9+/-2.9 microg/dl, 1.3+/-0.7 microg/dl, respectively) than those in both HSK cattle (n=5) (64.9+/-47.6 ng/ml, 97.8+/-40.7 ng/dl, 4.1+/-2.1 microg/dl and 1.8+/-1.1 microg/dl, respectively), and the controls (n=6) (314.7+/-197.2 ng/ml, 140.2+/-21.3 ng/dl, 5.8+/-1.7 microg/dl, and 3.0+/-1.4 microg/dl, respectively). The area under the concentration curve of growth hormone (GH-AUC 0-600 min) in MHO cattle (22210+/-18951 ng.min/ml) tended to be greater than those in HSK (7887+/-6340 ng.min/ml) and the controls (2811+/-1275 ng.min/ml). MHO cattle showed a high GH-AUC0-600 min in contrast to a low serum IGF-1 concentration, as well as lower serum T3, T4, and cortisol concentrations. HSK cattle exhibited the same secretory patterns, but much more moderately. Growth retardation in Japanese black cattle exhibits some variations based on pedigree.

  8. Diagnostic and Prognostic Significance of Lipid Profiles in Holstein Dairy Cattle with Displaced Abomasum: Before and After Surgical Operation

    Directory of Open Access Journals (Sweden)

    Arafat Khalphallah

    2016-01-01

    Full Text Available The study aimed to describe the pattern of changes in clinical findings and lipid metabolism profiles in dairy cattle with displacement of the abomasum (DA from day 0 until day 30 after operation. The study was conducted on DA cattle (n= 25 belonged to dairy farms in Hokkaido area, Japan. Cows were examined and sampled at days 0 (operation, 7 and 30. They were clinically and biochemically examined to estimate BCS and many serum biochemical constituents such as lecithin:cholesterol acyltransferase (LCAT and apolipoprotein B-100 (apoB-100, β-hydroxybutyric acid (BHBA, non-esterified fatty acids (NEFAs and aspartate amino transferase (AST. Based on blood BHBA at day 0, DA cows were classified into three categories; DA only (<1.2 mmol/l, DA with subclinical ketosis (DA SCK (1.2-2.4 mmol/l and DA with clinical ketosis (DA CK (≥2.5 mmol/l. The changes in the pattern of serum biochemical constituents throughout this study indicated recovery of diseased cows and significant effect of surgical operation. Serum biochemical constituents returned to their physiological values indicating that these cows were restoring their normal physiological status. This was reflected through a significant (P<0.05 elevation of LCAT, apoB-100, and cholesterol and a significant (P<0.05 reduction in AST, NEFAs and BHBA (Not in DA group, in all DA groups particularly at day 30 when their values compared with those at day 0. The current study also recorded no remarkable changes (P>0.05 between the diseased groups except for NEFA and BHBA (at day 0 between DA group and the other two groups at any of the three sampling days.

  9. A survey of small-scale cattle farming systems in the North West ...

    African Journals Online (AJOL)

    Unknown

    A survey of small-scale cattle farming systems in the North West Province of. South Africa ... Abstract. A questionnaire-based survey was conducted among a purposive sample of 60 small-scale cattle farmers in the .... has a negative impact on pasture and soil resources and represents a potential threat to the sustainability of.

  10. Sulphur levels in saliva as an estimation of sulphur status in cattle: a validation study

    NARCIS (Netherlands)

    Dermauw, V.; Froidmont, E.; Dijkstra, J.; Boever, de J.L.; Vyverman, W.; Debeer, A.E.; Janssens, G.P.J.

    2012-01-01

    Effective assessment of sulphur (S) status in cattle is important for optimal health, yet remains difficult. Rumen fluid S concentrations are preferred, but difficult to sample under practical conditions. This study aimed to evaluate salivary S concentration as estimator of S status in cattle.

  11. Development and evaluation of microwave-assisted and ultrasound-assisted methods based on a quick, easy, cheap, effective, rugged, and safe sample preparation approach for the determination of bisphenol analogues in serum and sediments.

    Science.gov (United States)

    Song, Shanjun; Shao, Mingwu; Wang, Weihua; He, Yajuan; Dai, Xinhua; Wang, Huiyu; Liu, Liliang; Guo, Feng

    2017-12-01

    Microwave- and ultrasound-assisted methods based on a quick, easy, cheap, effective, rugged, and safe sample preparation approach followed by high-performance liquid chromatography with tandem mass spectrometry were developed for the simultaneous determination of eight bisphenol analogues in serum and sediment. The developed methods provided satisfactory extraction efficiency for the energy provided by microwaves and ultrasound. Compositions of commercial sorbents (primary secondary amine, MgSO4 , octadecyl-modified silica, and graphitized carbon black) were evaluated. The ultrasound-assisted method was suited for serum using primary secondary amine, MgSO4 , and octadecyl-modified silica as sorbents and a mixture of hexane and ethyl acetate as extraction solvent. The microwave-assisted method worked better for sediment with tetrahydrofuran and methanol as solvents and primary secondary amine, MgSO4 , octadecyl-modified silica, and graphitized carbon black as sorbents. Other experimental parameters, such as extraction temperature and time, were also optimized. The inter- and intraday relative standard deviations ranged from 2.7 to 5.5%. The limits of detection were between 0.1 and 1.0 ng/mL for serum and between 0.1 and 0.5 ng/g dry weight for sediment. The proposed methods were successfully applied to seven sediment and 20 human serum samples. The results showed that the developed methods were practical for the analysis and biomonitoring of bisphenols in sera and sediment. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Serum uric acid change and modification of blood pressure and fasting plasma glucose in an overall healthy population sample: data from the Brisighella heart study.

    Science.gov (United States)

    Cicero, Arrigo F G; Rosticci, Martina; Bove, Marilisa; Fogacci, Federica; Giovannini, Marina; Urso, Riccardo; D'Addato, Sergio; Borghi, Claudio

    2017-06-01

    Serum uric acid (SUA) is an emerging risk factor for incident hypertension and type 2 diabetes. It is less clear if changes in SUA are associated to different incidence in these main cardiovascular risk factors. From the cohort of the Brisighella Heart Study, we selected non-diabetic subjects that in 2008 were untreated with SUA-lowering drugs nor antihypertensive ones. Then we divided the subjects in four main groups: the ones that maintained their SUA level unchanged during the next 4 years, the ones that increased it >1 mg/dL without treatment, the ones that reduced it >1 mg/dL without drug treatment and the ones that reduced it >1 mg/dL with the continuous use of allopurinol. Compared with 2008, SBP significantly increased in subjects with worsened (and untreated) SUA level, while improved in subjects treated with allopurinol (p population. Key messages Serum uric acid (SUA) is an emerging risk factor for incident hypertension and type 2 diabetes. SUA improvement could positively influence the age-related worsening of SBP and FPG in general population.

  13. On-line concentration by field-enhanced sample injection with reverse migrating micelles in micellar electrokinetic capillary chromatography for the analysis of coumarins from traditional Chinese medicine and human serum.

    Science.gov (United States)

    Jiang, Ting-Fu; Lv, Zhi-Hua; Wang, Yuan-Hong; Yue, Mei-E

    2010-06-01

    In this work, a simple, reproducible and sensitive micellar electrokinetic chromatography method was developed for the separation and determination of three coumarins, imperatorin (IM), isoimperatorin (IO) and osthole (OS) from traditional Chinese medicine and human serum. Field-enhanced sample injection with reverse migrating micelles was used for on-line concentration of the coumarins. The optimum buffer contained 50 mM H(3)PO(4), 160 mM sodium dodecyl sulfate, 20% acetonitrile and 15% 2-propanol, and the pH of buffer was 2.0. The sample solution was diluted with water containing 5 mM sodium dodecyl sulfate and injected for 15 s with -8 kV after injection of 2 s water plug. The effects of concentrations of sodium dodecyl sulfate and organic modifier, the sample matrix, the injection time of water plug, the injection voltage and injection time of sample on the separation and stacking efficiency were investigated. Under the optimum conditions, the analytes were well separated and by optimizing the stacking conditions, about 93, 195 and 136 fold improvement in the detection sensitivity was obtained for IM, IO and OS. The contents of three coumarins in Angelica dahurica Benth, Radix Angelicae Pubescentis and Fructus Cnidii were successfully determined with satisfactory repeatability and recovery. The possibilities of using this method for the determination of three coumarins in spiked human serum were also tested. Copyright 2009 John Wiley & Sons, Ltd.

  14. Evaluation of Genetic Diversity, Population Structure, and Relationship Between Legendary Vechur Cattle and Crossbred Cattle of Kerala State, India.

    Science.gov (United States)

    Radhika, G; Aravindakshan, T V; Jinty, S; Ramya, K

    2017-03-30

    The legendary Vechur cattle of Kerala, described as a very short breed, and the crossbred (CB) Sunandini cattle population exhibited great phenotypic variation; hence, the present study attempted to analyze the genetic diversity existing between them. A set of 14 polymorphic microsatellites were chosen from FAO-ISAG panel and amplified from genomic DNA isolated from blood samples of 30 Vechur and 64 unrelated crossbred cattle, using fluorescent labeled primers. Both populations revealed high genetic diversity as evidenced from high observed number of alleles, Polymorphic Information Content and expected heterozygosity. Observed heterozygosity was lesser (0.699) than expected (0.752) in Vechur population which was further supported by positive FIS value of 0.1149, indicating slight level of inbreeding in Vechur population. Overall, FST value was 0.065, which means genetic differentiation between crossbred and Vechur population was 6.5%, indicating that the crossbred cattle must have differentiated into a definite population that is different from the indigenous Vechur cows. Structure analysis indicated that the two populations showed distinct differences, with two underlying clusters. The present study supports the separation between Taurine and Zebu cattle and throws light onto the genetic diversity and relationship between native Vechur and crossbred cattle populations in Kerala state.

  15. Enzyme-Linked Immunosorbent Assay (Elisa) Based Detection of Antibodies to Mycoplasma bovis in Cattle Naturally Infected with Haemoparasites in Institutional farms in Sokoto State, Nigeria

    OpenAIRE

    F.M. Tambuwal; L. Stipkovits; G.O. Egwu; A.U. Junaidu; M.B. Abubakar and U.A. Turaki

    2011-01-01

    This was a cross-sectional study involving cattle from four (4) institutional farms (Prison farm, Livestock Investigation and Breeding Centre (LIBC), Usmanu Danfodiyo University Teaching and Research (UDUTRF) and Kebbe Cattle Ranch (KCR) in Sokoto state, Nigeria. A total of 62 cattle comprising 49 females and 13 males were randomly selected and bled from a total population of 205. The cattle sampled were local breeds comprising Gudali, Rahaji, White-Fulani and their crosses. They were aged 1-...

  16. Seneciosis in cattle associated with photosensitization

    Directory of Open Access Journals (Sweden)

    Paula R. Giaretta

    2014-05-01

    Full Text Available Senecio spp. poisoning is the main cause of cattle mortality in the central region of Rio Grande do Sul. This paper reports an outbreak of seneciosis in cattle with high prevalence of photosensitization, where 83 out of 162 cows (51.3% presented this clinical sign. The outbreak occurred in September 2013, affecting adult cows that were held in a 205 hectare-pasture from April to October 2013 with abundant Senecio brasiliensis infestation. Main clinical signs were weight loss, excessive lacrimation or mucopurulent ocular discharge, nasal serous discharge, ventral diphteric glossitis, crusts in the nose, teats, dorsum of ears, and vulva. Liver biopsy was performed in all the cows under risk; the histopathological findings in the liver biopsies consisted of fibrosis, megalocytosis, and biliary ductal proliferation and were present in 73.4% of the biopsied animals. Six cows had increased serum activity of gamma glutamyl transferase. Three affected cows were necropsied. The main necropsy findings were a hard liver, distended gall bladder, edema of the mesentery and abomasum. Liver histological changes in the necropsied cows were similar to those of the biopsied livers. Spongiosis was detected in the brain of necropsied cows and is characteristic of hepatic encephalopathy.

  17. Clinical utility of HCV core antigen detection and quantification using serum samples and dried blood spots in people who inject drugs in Dar-es-Salaam, Tanzania

    Directory of Open Access Journals (Sweden)

    Zameer Mohamed

    2017-01-01

    Results: Out of 153 HCV-seropositive individuals, 65 (42.5% and 15 (9.8% were co-infected with HIV (41 (63% were on anti-retroviral therapy (ARVs and hepatitis B respectively. In total, 116 were viraemic, median viral load of 5.7 (Interquartile range (IQR; 4.0–6.3 log iU/ml (75 (68.2% were genotype 1a, 35 (31.8% genotype 4a. The median alanine transaminase (ALT (iU/l, aspartate transaminase (AST (iU/l and gamma-glutamyl transferase (GGT (iU/l were 35 (IQR; 23–51, 46 (32–57 and 69 (35–151 respectively. For the quantification of HCV RNA, serum HCVcAg had a sensitivity at 99.1% and a specificity at 94.1%, with an area under the receiver operating curve (AUROC at 0.99 (95% CI 0.98–1.00. DBS HCVcAg had a sensitivity of 76.1% and a specificity of 97.3%, with an AUROC of 0.87 (95% CI 0.83–0.92. HCVcAg performance did not differ by HIV co-infection or HCV genotype.Conclusions: Our study suggests that HCVcAg testing in serum is an excellent alternative to HCV polymerase chain reaction in Africa. Although HCVcAg detection and quantification in DBS has a reduced sensitivity, its specificity and accuracy are good and it could therefore be used for scaling up HCV testing and care in resource-limited African settings.

  18. Metabolomic and Lipidomic Analysis of Serum Samples following Curcuma longa Extract Supplementation in High-Fructose and Saturated Fat Fed Rats.

    Science.gov (United States)

    Tranchida, Fabrice; Shintu, Laetitia; Rakotoniaina, Zo; Tchiakpe, Léopold; Deyris, Valérie; Hiol, Abel; Caldarelli, Stefano

    2015-01-01

    We explored, using nuclear magnetic resonance (NMR) metabolomics and fatty acids profiling, the effects of a common nutritional complement, Curcuma longa, at a nutritionally relevant dose with human use, administered in conjunction with an unbalanced diet. Indeed, traditional food supplements have been long used to counter metabolic impairments induced by unbalanced diets. Here, rats were fed either a standard diet, a high level of fructose and saturated fatty acid (HFS) diet, a diet common to western countries and that certainly contributes to the epidemic of insulin resistance (IR) syndrome, or a HFS diet with a Curcuma longa extract (1% of curcuminoids in the extract) for ten weeks. Orthogonal projections to latent structures discriminant analysis (OPLS-DA) on the serum NMR profiles and fatty acid composition (determined by GC/MS) showed a clear discrimination between HFS groups and controls. This discrimination involved metabolites such as glucose, amino acids, pyruvate, creatine, phosphocholine/glycerophosphocholine, ketone bodies and glycoproteins as well as an increase of monounsaturated fatty acids (MUFAs) and a decrease of n-6 and n-3 polyunsaturated fatty acids (PUFAs). Although the administration of Curcuma longa did not prevent the observed increase of glucose, triglycerides, cholesterol and insulin levels, discriminating metabolites were observed between groups fed HFS alone or with addition of a Curcuma longa extract, namely some MUFA and n-3 PUFA, glycoproteins, glutamine, and methanol, suggesting that curcuminoids may act respectively on the fatty acid metabolism, the hexosamine biosynthesis pathway and alcohol oxidation. Curcuma longa extract supplementation appears to be beneficial in these metabolic pathways in rats. This metabolomic approach highlights important serum metabolites that could help in understanding further the metabolic mechanisms leading to IR.

  19. Assessment of CCL2 and CXCL8 chemokines in serum, bronchoalveolar lavage fluid and lung tissue samples from dogs affected with canine idiopathic pulmonary fibrosis.

    Science.gov (United States)

    Roels, Elodie; Krafft, Emilie; Farnir, Frederic; Holopainen, Saila; Laurila, Henna P; Rajamäki, Minna M; Day, Michael J; Antoine, Nadine; Pirottin, Dimitri; Clercx, Cecile

    2015-10-01

    Canine idiopathic pulmonary fibrosis (CIPF) is a progressive disease of the lung parenchyma that is more prevalent in dogs of the West Highland white terrier (WHWT) breed. Since the chemokines (C-C motif) ligand 2 (CCL2) and (C-X-C motif) ligand 8 (CXCL8) have been implicated in pulmonary fibrosis in humans, the aim of the present study was to investigate whether these same chemokines are involved in the pathogenesis of CIPF. CCL2 and CXCL8 concentrations were measured by ELISA in serum and bronchoalveolar lavage fluid (BALF) from healthy dogs and WHWTs affected with CIPF. Expression of the genes encoding CCL2 and CXCL8 and their respective receptors, namely (C-C motif) receptor 2 (CCR2) and (C-X-C motif) receptor 2 (CXCR2), was compared in unaffected lung tissue and biopsies from dogs affected with CIPF by quantitative PCR and localisation of CCL2 and CXCL8 proteins were determined by immunohistochemistry. Significantly greater CCL2 and CXCL8 concentrations were found in the BALF from WHWTs affected with CIPF, compared with healthy dogs. Significantly greater serum concentrations of CCL2, but not CXCL8, were found in CIPF-affected dogs compared with healthy WHWTs. No differences in relative gene expression for CCL2, CXCL8, CCR2 or CXCR2 were observed when comparing lung biopsies from control dogs and those affected with CIPF. In affected lung tissues, immunolabelling for CCL2 and CXCL8 was observed in bronchial airway epithelial cells in dogs affected with CIPF. The study findings suggest that both CCL2 and CXCL8 are involved in the pathogenesis of CIPF. Further studies are required to determine whether these chemokines might have a clinical use as biomarkers of fibrosis or as targets for therapeutic intervention. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Multi-antigen print immunoassay for seroepidemiological surveillance of bovine tuberculosis on Indian cattle farms

    Directory of Open Access Journals (Sweden)

    Maroudam Veerasami

    2012-09-01

    Full Text Available Bovine tuberculosis caused by Mycobacterium bovis is a zoonotic disease that is responsible for significant economic losses in many countries. The standard diagnostic method, the tuberculin test (TST that is used in control programmes has serious shortcomings and, given the complex nature and the economic impact of the disease, a number of other diagnostic methods have been examined. The authors have attempted to characterise antibody response using the multi-antigen print immunoassay (MAPIA. A total of 511 serum samples were collected from farms in India on which bovine tuberculosis was prevalent and on farms with low incidence. These were tested using the MAPIA against a panel of five defined M. bovis recombinant antigens and two purified protein derivatives (bovine PPD and avian PPD to study the seroprevalence of the disease on Indian cattle farms. Results indicated that the fusion protein of antigen CFP-10:MPB83 showed a positive response in 142 out of 298 serum samples from tuberculosis-prevalent farms, thereby indicating the serological dominance of the proteins post infection. The antigen selected could be used further in the development of a simple, rapid and accurate serological diagnostic test, paired with TST, for use in bovine tuberculosis control programmes.

  1. Detection of Theileria lestoquardi cross infection in cattle with clinical theileriosis in Iran.

    Science.gov (United States)

    Jalali, Seyedeh Missagh; Jolodar, Abbas; Rasooli, Aria; Darabifard, Ameneh

    2016-12-01

    Theileriosis caused by Theileria lestoquardi (malignant ovine theileriosis) in sheep and Theileria annulata (tropical theileriosis) in cattle is an important hemoprotozoal tick-borne disease in Iran. Due to major biologic and phylogenic similarities of these two species, this study was carried out to investigate the occurrence of natural infections with T.lestoquardi and T.annulata in cattle with clinical theileriosis in Ahvaz, southwest Iran. Fifty one cattle were selected based on clinical signs of theileriosis and confirmation by microscopic examination of blood smears. Blood samples were collected from each animal and hematologic and microscopic examinations were performed. Theileria piroplasmic forms were detected in all affected cattle. Pale mucous membranes (43.14%), icterus (11.76%) and fever (70.6%) were also observed. PCR-RFLP analysis revealed T. annulata infection in all tested cattle while coinfections with T. lestoquardi were found in two samples (3.92%). All sampled cattle including the two with mixed species Theileria infection were anemic. This is the first report of Theileria species cross infections in cattle with clinical theileriosis in Iran. It can be concluded that cattle can be infected with both pathogenic Theileria species, T. lestoquardi and T. annulata which can be an important issue in the epidemiology and spread of ovine malignant theileriosis.

  2. [Determination of bilirubin in capillary plasma by a direct photometric method (DPM, bilirubinometer) and the chemical determination of bilirubin in the bilirubin determination (2,5-dichlorophenyldiazonium method) in serum of venous blood samples].

    Science.gov (United States)

    Schlebusch, H; Liappis, N; Niesen, M

    1988-01-01

    The determination of bilirubin in serum was performed by the 2.5-dichlorphenyldiazonium method (DPD) and in capillary plasma by the direct photometric method (DPM). Both methods showed a good precision and accuracy. The investigation was carried out in 135 samples with a bilirubin concentration up to 25 mg/dl. The comparison of the two methods in 62 samples with a bilirubin concentration up to 10 mg/dl showed a correlation coefficient of r = 0.862 and in 73 samples with a bilirubin concentration between 10 and 25 mg/dl a correlation coefficient of r = 0.893. In 29 cases (21.5%) we found differences between the two methods of 1.5-4.0 mg/dl. Most of them were in the critical higher range. Discussion of the DPD and DPM methods.

  3. Mycobacterium avium subsp. paratuberculosis Antibody Response, Fecal Shedding, and Antibody Cross-Reactivity to Mycobacterium bovis in M. avium subsp. paratuberculosis-Infected Cattle Herds Vaccinated against Johne's Disease

    Science.gov (United States)

    Hovingh, Ernest; Linscott, Rick; Martel, Edmond; Lawrence, John; Wolfgang, David; Griswold, David

    2014-01-01

    Vaccination for Johne's disease with killed inactivated vaccine in cattle herds has shown variable success. The vaccine delays the onset of disease but does not afford complete protection. Johne's disease vaccination has also been reported to interfere with measurements of cell-mediated immune responses for the detection of bovine tuberculosis. Temporal antibody responses and fecal shedding of Mycobacterium avium subsp. paratuberculosis, the causative agent of Johne's disease, were measured in 2 dairy cattle herds using Johne's disease vaccine (Mycopar) over a period of 7 years. Vaccination against Johne's disease resulted in positive serum M. avium subsp. paratuberculosis antibody responses in both herds, and the responses persisted in vaccinated cattle up to 7 years of age. Some vaccinated animals (29.4% in herd A and 36.2% in herd B) showed no serological reactivity to M. avium subsp. paratuberculosis. M. avium subsp. paratuberculosis-specific antibody responses were also detected in milk from Johne's disease-vaccinated animals, but fewer animals (39.3% in herd A and 49.4% in herd B) had positive results with milk than with serum samples. With vaccination against M. avium subsp. paratuberculosis, fecal shedding in both dairy herds was reduced significantly (P < 0.001). In addition, when selected Johne's disease-vaccinated and -infected animals were investigated for serological cross-reactivity to Mycobacterium bovis, no cross-reactivity was observed. PMID:24623626

  4. Comparison of serum pools and oral fluid samples for detection of porcine circovirus type 2 by quantitative real-time PCR in finisher pigs

    DEFF Research Database (Denmark)

    Nielsen, Gitte Blach; Nielsen, Jens Peter; Haugegaard, John

    2018-01-01

    PCR of paired samples at the pen level of pools of sera (SP) from 4 to 5 pigs and the collective oral fluid (OF) from around 30 pigs corresponding to one rope put in the same pen. Pigs in pens of 2 finishing herds were sampled by cross-sectional (Herd 1) and cross-sectional with follow-up (Herd 2) study designs...

  5. Assessment of the probability of introduction of bovine tuberculosis to Danish cattle farms via imports of live cattle from abroad and immigrant workers.

    Science.gov (United States)

    Foddai, Alessandro; Nielsen, Liza Rosenbaum; Krogh, Kaspar; Alban, Lis

    2015-12-01

    Denmark has been recognized as officially free (OTF) from bovine tuberculosis (bTB) since 1980. In this study, we estimated the annual probability (PIntro) of introducing Mycobacterium bovis into the Danish cattle population, through (a) imports of cattle and (b) foreign personnel working in Danish cattle herds. Data from 2000 to 2013 with date, number and origin of imported live cattle were obtained from the Danish Cattle Federation. Information on immigrants working in Danish cattle herds was obtained through a questionnaire sent by email to a sample of Danish cattle farmers (N=460). Inputs obtained from data analysis, expert opinion, the questionnaire and literature were fed into three stochastic scenario tree models used to simulate the effect of import trade patterns, and contact between immigrant workers and cattle. We also investigated the opportunity of testing animals imported from OTF countries by tuberculin skin test and animals from non-OTF countries by interferon-γ test (IFN-γ), exemplified by using year 2009 where the number of imported animals was higher than usual. Results showed that PIntro is driven mainly by importation of live cattle. The combined median annual probability of introducing M. bovis into the Danish cattle population by either imported live cattle or infectious immigrant workers, ranged from 0.3% (90% prediction interval (P.I.): 0.04%:1.4%) in 2001 to 4.9% (90% P.I.: 0.6%; 19.2%) in 2009. The median of the median PIntro estimates from the 14 years was 0.7% (median of 90% P.I.: 0.08%; 3.5%). Hence, on average, at least one introduction each 143 years could be expected, if the annual number of imported animals does not change remarkably in the future. If the number of imported animals increases, compared to the years we analyzed, additional testing of imported cattle might be considered. For example, in 2009, PIntro would have been reduced from 4.9% to 0.8% (90% P.I.: 0.1%; 4.7%) if animals from OTF countries had been tested with

  6. Investigation of haemoglobin polymorphism in Ogaden cattle

    Directory of Open Access Journals (Sweden)

    Sanjoy Kumar Pal

    2014-04-01

    Full Text Available Background and Aim: The Ogaden cattle is one among the tropical cattle breeds (Bos indicus widely distributed in eastern and south eastern part of Ethiopia. The breed has been evolved in arid and semi arid agro-ecological setup, but later on distributed and adapted to the wide agro-ecological zones. Because of its multi-purpose role, the Ogaden cattle have been used for milk, beef, and income generation. Information on the inherent genetic diversity is important in the design of breeding improvement programmes, making rational decisions on sustainable utilization and conservation of Animal Genetic Resources. Limited information is available about genetic variation of Ogaden breed at molecular level. The present investigation was aimed to study the biochemical polymorphism at the Hemoglobin (Hb locus. Materials and Methods: Blood samples collected from 105 Ogaden cattle maintained at Haramaya beef farm by jugular vein puncture were subjected to agarose gel electrophoresis [pH range 8.4-8.5] to study the polymorphic activities of haemoglobin. Results: Three types of phenotypes were detected i.e. a slow moving (AA band, fast moving (BB band and a combination of slow + fast moving bands (AB. The frequency of the fast moving band was less [13 (12.3%] than the slow moving band [57 (54.2%]. Both slow & fast moving phenotype was observed in 35 (33.3% animals. The gene frequency of HBA allele was 0.709 and that of HBB allele 0.291. Conclusion: The distribution of phenotypes was in agreement with codominant single gene inheritance. The Chi-square (χ2 test revealed that the population is under Hardy-Weinberg equilibrium.

  7. Seroprevalence of Neospora caninum Infection in Dairy Cattle in West of Iran

    Directory of Open Access Journals (Sweden)

    Hassan NAYEBZADEH

    2015-07-01

    Full Text Available The Neospora caninum parasite causes abortion in cattle in virtually all parts of the world with enormous economic consequences. The purpose of this study was to determine the seroprevalence of antibodies of Neospora caninum in dairy cattle in Lorestan Province, west of Iran. A total of 347 dairy cows were randomly selected. The serum of each case was analyzed for the possibility of the presence of antibody against N. caninum antigen, using the commercial kit: ELISA. The results of the ELISA test indicated that from 347 dairy cattle examined, the antibodies to N. caninum were found in 34 (9.8%. The percentage of seropositive aborted cattle was 13.33%. This study also indicated that there was no significant relationship between seropositivity and such factors as the age, breed, and abortion history of the cattle. Moreover, no significant relationship between seroprevalence of infection among rural and industrial cows was found. The neosporosis could be one of the possible causes of abortion in cattle. Further studies are recommended to determine the relationship between this parasite and the occurrence of abortion in cattle in the province of Lorestan.

  8. The nasopharyngeal microbiota of feedlot cattle that develop bovine respiratory disease.

    Science.gov (United States)

    Holman, Devin B; McAllister, Tim A; Topp, Edward; Wright, André-Denis G; Alexander, Trevor W

    2015-10-22

    Bovine respiratory disease is the major cause of morbidity and mortality in feedlot cattle. The objective of this study was to compare the nasopharyngeal bacterial microbiota of healthy cattle and cattle treated for BRD in a commercial feedlot setting using a high-density 16S rRNA gene microarray (Phylochip). Samples were taken from both groups of animals (n=5) at feedlot entry (day 0) and ≥60 days after placement. Cattle diagnosed with BRD had significantly less bacterial diversity and fewer OTUs in their nasopharynx at both sampling times. The predominant phyla in both groups were Proteobacteria and Firmicutes. The relative abundance of the phylum Actinobacteria was lower in cattle treated for BRD. At the family-level there was a greater relative abundance (Pbovine nasopharyngeal microbiota is relatively unstable during the first 60 days in the feedlot. Copyright © 2015. Published by Elsevier B.V.

  9. Keterkaitan antara Turbiditas Serum dan Laju Endap Darah dengan Kerusakan Hati pada Sapi Bali

    Directory of Open Access Journals (Sweden)

    Iwan Harjono Utama

    2010-09-01

    Full Text Available This research was aimed to observe serum turbidity and blood sedimentation rate (ESR as a predictorof hepatic damage in bali cattle. Two hundred whole blood and sera from 80 male and 120 female bullswere sampled from Mambal abbatoir, Badung, Bali. All blood were examined for their ESR and sera werefor their turbidity using ZnSO4 solution, besides we observed some hepatic damages pathology anatomicallywithout incision such as : internal bleeding, formation of connective tissue, swelling, and bile-ductenlargement. all of those damages were scaled graded from 0 (without abnormality to 4 (more than 75%liver surface has abnormalities. Results showed all 86% bulls (male 32% and female 54% have their seraturbidity ranged from 1,01-2,00, besides, 89% bulls (45% male and 44% female have their ESR rangedfrom 3-8 mm in 24 hours. Most of liver abnormalities were : swelling (58%, bile-duct enlargement (50,5%,connective tissue formation (73%, and bleeding (59% most of them were falling in scale 1 (less than 25%of liver surface area. Also, ESR has positive correlation (P<0,05 with connective tissue formation andserum turbidity value could be used to predict internal bleeding and connective tissue formation. It couldbe concluded ESR value could be used as a predictor of connective tissue formation and serum turbidityvalue could be used as a predictor for internal bleeding and connective tissue formation in bali cattle.

  10. Serological surveillance of bluetongue virus in cattle in central Iran

    Directory of Open Access Journals (Sweden)

    Vahid Noaman

    2013-06-01

    Full Text Available The aim of this study was to evaluate the seroprevalence and distribution of antibodies to the bluetongue virus (BTV among dairy Holstein cattle of central Iran. From September 2010 to August 2011, 892 blood samples from Holstein dairy cattle were collected from healthy animals. Blood samples were divided according to type of farm (industrial and non-industrial, season (warm and cold, location (North, South, East, and West, cattle production groups (calf, heifer, dairy and dry and age groups (under 6 months, 6 months-2 years and over 2 years. The sera were screened using a commercially competitive enzyme-linked immunosorbent assay (c-ELISA kit. Twenty-four sera (2.69 % were found to be positive for BTV. Bluetongue virus seroprevalence was significantly higher (χ2 = 8.29, df = 3, p 2 years showed a relatively higher seroprevalence, but the difference was not statistically significant (p = 0.06. No statistically significant difference in BTV seroprevalence was noted between farming systems, seasons and cattle production groups (p > 0.05. The results demonstrate that the seroprevalence of BTV is low in cattle from the Isfahan province, central Iran. Further studies are needed to determine the serotypes and vectors of BTV in the central region of Iran.

  11. Seroepidemiological study of bovine respiratory viruses (BRSV, BoHV-1, PI-3V, BVDV, and BAV-3) in dairy cattle in central region of Iran (Esfahan province).

    Science.gov (United States)

    Shirvani, Edris; Lotfi, Mohsen; Kamalzadeh, Morteza; Noaman, Vahid; Bahriari, Masumeh; Morovati, Hasan; Hatami, Alireza

    2012-01-01

    Respiratory diseases in calves are responsible for major economic losses in both beef and dairy production. Several viruses, such as bovine respiratory syncytial virus (BRSV), bovine herpes virus-1 (BoHV-1), bovine parainfluenza virus-3 (BPI-3V), bovine viral diarrhea virus (BVDV), and bovine adenoviruses (BAV), are detected in most clinical cases with respiratory signs. The aim of this study is to define seroprevalences of five major viral causes of bovine respiratory infections in cattle in central region of Iran (Esfahan province). The population targeted was 642 dairy cows (Holstein-Friesian) from 25 farms. Samples of blood serum from female cattle were examined. Sera were tested by commercial ELISA kits to detect antibody against BRSV, BoHV-1, BPI-3V, BVDV, and BAV-3. The results were analyzed by Chi-square test. In the present study, seroprevalences of BRSV, BoHV-1, PI3V, BVDV, and BAV-3 were 51.1%, 72%, 84.4%, 49.2%, and 55.6%, respectively. The present study shows that infections of bovine respiratory viruses are very common in cattle in Esfahan.

  12. Sero-prevalence of Taenia spp. infections in cattle and pigs in rural farming communities in Free State and Gauteng provinces, South Africa.

    Science.gov (United States)

    Tsotetsi-Khambule, A M; Njiro, S; Katsande, T C; Thekisoe, O M M; Harrison, L J S

    2017-08-01

    The aim of this study was to determine sero-prevalence of bovine and porcine cysticercosis in cattle and pigs in rural farming communities in Free State and Gauteng Provinces, Republic of South Africa. Blood samples were collected for a period of twelve months from live cattle (n=1315; 1159) and pigs (n=436; 240) and the serum extracted and stored before analysis by a monoclonal antibody based (HP10) antigen detection ELISA. Results revealed a generally high sero-prevalence and wide distribution throughout the two provinces with Free State having a higher sero-prevalence in both cattle and pigs (23% and 34%) than Gauteng province (15% and 14%). Consumption of infected meat that is either not inspected/missed at meat inspection; poor livestock management practices and limited sanitation in rural communities might have contributed to the occurrence of Taenia spp. infections in the two provinces. It is therefore, recommended that cysticercosis status of animals be established before slaughter. This would assist in ensuring that infected animals are not slaughtered for human consumption or zoonosis preventive measures are taken. Furthermore, public awareness programs on life cycles of T. saginata, T. solium and T. hydatigena and the use of more sensitive diagnostic tools are recommended as part of effective control strategies against taeniid infections. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Economic assessment of Ostertagia ostertagi and Fasciola hepatica infections in dairy cattle herds in Germany using Paracalc®.

    Science.gov (United States)

    Fanke, Jane; Charlier, Johannes; Steppin, Torsten; von Samson-Himmelstjerna, Georg; Vercruysse, Jozef; Demeler, Janina

    2017-06-15

    The aim of the current study was to estimate economic costs of Ostertagia ostertagi and Fasciola hepatica infections in dairy cattle herds in Germany using the online calculation programme Paracalc ® . Following a questionnaire, survey data were available from 464 farms in 14 federal states. On those farms bulk tank milk (BTM) samples and additionally up to six serum samples collected from first season grazing calves were analysed, using a commercially available ELISA (Boehringer Ingelheim SVANOVA Biotech AB, Uppsala, Sweden), an in-house ELISA (F. hepatica) and an in-house serum pepsinogen test. In total, samples obtained from 344 farms were included in the analysis since those were the only farms with complete questionnaires. Median costs per farm and year were estimated for gastrointestinal (GI) nematode infections (€721.38) and F. hepatica infection (€565.61). Decreases in milk yield in multiparous cows were the major reason for annual production losses due to GI nematodes (€13.33 per cow) and F. hepatica infections (€7.95 per cow), which was followed by annual costs for anthelmintic treatment against GI nematode infections in adult cows (€10.00 per cow) and F. hepatica infection associated annual costs due to repeated artificial insemination (€10.13 per cow) and prolonged calving intervals (€9.40 per cow). The study demonstrated that if all required information is provided, the Paracalc ® tool can assist to identify productions losses in dairy cattle herds due to helminth infections and to optimise farm economics in Germany. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Prevalence and risk factors for Leptospira spp. in cattle herds in the south central region of Paraná state

    Directory of Open Access Journals (Sweden)

    Vanessa Y Hashimoto

    2012-02-01

    Full Text Available The aim of this study was to determine the prevalence of anti-Leptospira spp. antibodies and the risk factors for Leptospira spp. infection in breeding cattle herds in the south central region of Paraná state. It was based on the statistic delineation/serological samples and information regarding the selected farms employed in the study of bovine brucellosis for Paraná state in the context of National Program for Control and Eradication of Brucellosis and Tuberculosis. A total of 1.880 females aged >24 months from 274 non vaccinated herds were studied. Serum samples were tested for antibodies against Leptospira spp. using microscopic agglutination test (MAT with 22 Leptospira serovars. The epidemiological questionnaire was applied on all the selected farms and aimed to obtain epidemiological data. Hundred eighty one of 274 herds were positive for Leptospira spp./presenting prevalence of positive herds of 66.06% (IC95%=60.12-71,65%. Presence of >43 cattle (OR=3.120; IC=1.418-6.867/animal purchase (OR=2.010; IC=1.154-3.500/rent of pastures (OR=2.925; IC=1.060-8.068 and presence of maternity paddock (OR=1.981; IC=1,068-3,676 were identified as risk factors for leptospirosis due to any serovar in the multivariate logistic regression. Risk factors for leptospirosis due to serovar Hardjo were presence of >43 cattle (OR=3.622; IC=1.512-8,677/animal purchase (OR=3.143; IC=1.557-6.342/rent of pastures (OR=4.070; IC=1.370-12.087 and presence of horses (OR=2.981; IC=1.321-6.726. These results indicate that Leptospira spp. infection is widespread in the south central region of Paraná state and that factors related to the herd characteristic and management are associated with the infection.

  15. Comparison of the Q-fever complement fixation test and two commercial enzyme-linked immunosorbent assays for the detection of serum antibodies against Coxiella burnetti (Q-fever) in ruminants : recommendations for use of serological tests on imported animals in New Zealand.

    Science.gov (United States)

    Kittelberger, R; Mars, J; Wibberley, G; Sting, R; Henning, K; Horner, G W; Garnett, K M; Hannah, M J; Jenner, J A; Piggott, C J; O'Keefe, J S

    2009-10-01

    To make valid recommendations on the use of serological test methods for the detection of serum antibodies in ruminants against Coxiella burnetii (Q-fever), by comparing the performance of the complement fixation test (CFT) and two ELISA, and by identifying reasons for discrepancies between the test methods. A total of 73 serum samples from infected cattle, 69 from infected goats, and 100 samples from non-infected cattle and 57 samples from non-infected sheep, as well as 95 samples from infected cattle herds (mix of seropositive and seronegative samples), were tested using the CFT, the IDEXX ELISA (I-ELISA) and the Pourquier ELISA (P-ELISA). A mixed panel of 12 serum samples from sheep from inter-laboratory proficiency testing (proficiency panel) was also tested using the CFT and both ELISA, and further investigated using IgG- and IgM-specific ELISA. Generally, the two commercial ELISA were more sensitive than the CFT for the detection of infected ruminants. Good agreement between ELISA for positive and negative results was found for samples from the infected herd, while results for the positive panels varied between the two ELISA. For the total of the positive serum panels, the I-ELISA detected 95% of samples as positive or suspicious, while the P-ELISA detected only 81%. In the P-ELISA, more samples were considered suspicious (18%) than in the I-ELISA (14%). All sera from non-infected sheep and cattle tested negative in the serological test methods employed, except for one positive sample from a sheep in the P-ELISA. Further investigation revealed that a CFT-positive but ELISA-negative result was due to high IgM and low IgG reactivity. The two commercial ELISA were more sensitive than the CFT in all panels from infected ruminants. However, they could only detect IgG. The I-ELISA should be the serological test method of choice for cattle, sheep and goats for import testing of animals into New Zealand because it was more sensitive than the P-ELISA and was equally

  16. Vitamin deficiencies in cattle.

    Science.gov (United States)

    Frye, T M; Williams, S N; Graham, T W

    1991-03-01

    Deficiencies of vitamins A, D, K, E and thiamin can cause severe limitations in beef production. In particular, vitamin A and E can be common causes of lost profit, secondary to limitations of reproductive and growth potential. Prolonged dry periods will reduce available A and E in pasture forage, as can ensiling and prolonged storage of harvested feedstuffs. Polioencephalomalacia is a thiamin responsive disorder, associated with high concentrate feeding and lush pastures. Antimetabolites, such as amprolium, will cause thiamine deficiency when fed in excess. Recent information has shown improved performance with supplemental beta carotene and niacin. The positive responses in reproductive performance, noted with cattle fed supplemental beta carotene, was independent of vitamin A. Supplementation of vitamins above National Research Council recommendations can be justified. However, proper evaluation of feed and animal status, and documentation of a response to supplementation is necessary before diagnosing deficiencies of specific nutrients.

  17. Serological response of cattle to Brucella allergen after repeated intradermal applications of this allergen

    NARCIS (Netherlands)

    Muskens, J.A.M.; Bercovich, Z.; Damen, C.P.R.M.

    1996-01-01

    A study was conducted to determine whether an allergen that has been prepared from a mucoid strain of Brucella abortus triggers a serum antibody response that interferes with the interpretation of serologic tests results. Fifteen cattle seronegative for Brucella antigen were tested with the SDTH

  18. Seroprevalence and risk factors for leptospirosis in cattle, sheep, and goats at consorted rearing from the State of Piauí, northeastern Brazil.

    Science.gov (United States)

    Campos, Ângela Piauilino; Miranda, Dayane Francisca Higino; Rodrigues, Huanna Waleska Soares; da Silva Carneiro Lustosa, Micherlene; Martins, Gustavo Henrique Chaves; Mineiro, Ana Lys Bezerra Barradas; Castro, Vanessa; Azevedo, Sérgio Santos; de Sousa Silva, Silvana Maria Medeiros

    2017-06-01

    Leptospirosis is an endemic disease in Latin America, caused by pathogenic bacteria of the genus Leptospira. It is considered one of the main causes responsible for the negative economic impact on global livestock by causing reproductive problems. The research aimed to determine the prevalence of leptospirosis in cattle, sheep, and goats at consorted rearing in the micro-region of Teresina, Piauí state, northeastern Brazil, as well as to identify prevalent serovars and risk factors associated with seroprevalence. Serum samples were analyzed in 336 sheep, 292 goats, and 253 cattle using microscopic agglutination test (MAT). Overall, 378 samples were positive to MAT, with seroprevalence of 42.9%. The prevalences in cattle, sheep, and goats were 50.5, 40.5, and 34.6%, respectively. All herds presented at least one seropositive animal; the Hardjo/Wolffi serovar association was the most common in cattle and Icterohaemorrhagiae in goats and sheep. Beef production (OR = 4.9), cattle herd over 35 animals (OR = 4.0), feeding on pasture (OR = 6.4), weir and/or stream as water source (OR = 2.1), and no veterinary services (OR = 2.9) were risk factors for cattle infection. For sheep, intensive management system (OR = 5.3), suspended slatted facilities (OR = 2.2), more than 20 sheep in reproductive age (OR = 1.9), and absence of deworming (OR = 3.5) were the risk factors, while for goats, the identified risk factors were sheep herd over 52 animals (OR = 1.9) and no veterinary services (OR = 1.8). We conclude that the infection was spreading in consorted herds in this region. Thus, it would be interesting and important to conduct educative activities to farmers on the economic impacts of this disease and the need of preventive and control strategies mainly focused on sanitary measures and animal handling.

  19. A highly sensitive and selective electrochemical determination of non-steroidal prostate anti-cancer drug nilutamide based on f-MWCNT in tablet and human blood serum sample.

    Science.gov (United States)

    Karthik, R; Sasikumar, R; Chen, Shen-Ming; Vinoth Kumar, J; Elangovan, A; Muthuraj, V; Muthukrishnan, P; Al-Hemaid, Fahad M A; Ajmal Ali, M; Elshikh, Mohamed S

    2017-02-01

    A novel electrochemical sensor based on the functionalized multiwalled carbon nanotube (f-MWCNT) was successfully developed for the sensitive and selective determination of non-steroidal prostate anti-cancer drug nilutamide in tablet and blood serum samples. The f-MWCNT was prepared by the simple reflux method and characterized by the scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), Raman spectroscopy, X-ray powder diffraction (XRD) and fourier transform infrared spectroscopy (FT-IR). Interestingly, the f-MWCNT was exhibited a superior electrocatalytic activity towards the anti-cancer drug nilutamide when compared with pristine MWCNT and unmodified electrodes. Besides, the electrochemical sensor was revealed an excellent current response for the determination of nilutamide with wide linear ranges (0.01-21μM and 28-535μM), high sensitivity (11.023 and 1.412μA μM-1cm2) and very low detection limit (LOD) 0.2nM. The developed electrochemical sensor was showed an excellent selectivity even in the presence of electrochemically active biological substances and nitro aromatic compounds. Moreover, it manifested a good reproducibility and stability. In addition, the f-MWCNT modified glassy carbon electrode (GCE) sensor was successfully applied for the detection of nilutamide in tablet and blood serum sample. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. Development of novel molecularly imprinted magnetic solid-phase extraction materials based on magnetic carbon nanotubes and their application for the determination of gatifloxacin in serum samples coupled with high performance liquid chromatography.

    Science.gov (United States)

    Xiao, Deli; Dramou, Pierre; Xiong, Nanqian; He, Hua; Li, Hui; Yuan, Danhua; Dai, Hao

    2013-01-25

    A novel composite imprinted material, on the basis of magnetic carbon nanotubes (MCNTs)-incorporated layer using gatifloxacin as a template, methacrylic acid as a functional monomer, and ethylene glycol dimethacrylate as a cross-linker, was successfully synthesized by a surface imprinting technique. Adsorption dynamics and a Scatchard adsorption model were employed to evaluate the adsorption process. The results showed that magnetic carbon nanotubes molecularly imprinted polymers (MCNTs@MIP) displayed a rapid dynamic adsorption and a high adsorption capacity of 192.7 μg/mg toward GTFX. Applied MCNTs@MIP as a sorbent, a magnetic solid phase extraction method coupled with high performance liquid chromatography (MSPE-HPLC) was developed for the determination of GTFX in serum samples. The recoveries from 79.1±4.8% to 85.3±4.2% were obtained. MCNTs@MIP can not only be collected and separated fast by external magnetic field but also have high surface-to-volume ratio, outstanding mechanical properties and specific recognition toward template molecule. In addition, the MCNTs@MIP could be regenerated, which could be used for five cycles with lost of less than 7.8% of its recovery on average. These analytical results of serum samples display that the proposed method based on MCNTs@MIP is applicable for fast and selective extraction of therapeutic agents from biological fluids. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Low ficolin-3 levels in early follow-up serum samples are associated with the severity and unfavorable outcome of acute ischemic stroke

    DEFF Research Database (Denmark)

    Fust, George; Fog, Lea Munthe; Illes, Zsolt

    2011-01-01

    papers demonstrated the significance of MBL in ischemic stroke, the role of ficolins has not been examined. METHODS: Sera were obtained within 12 hours after the onset of ischemic stroke (admission samples) and 3-4 days later (follow-up samples) from 65 patients. The control group comprised 100 healthy...... individuals and 135 patients with significant carotid stenosis (patient controls). The concentrations of ficolin-2 and ficolin-3, initiator molecules of the lectin complement pathway, were measured by ELISA methods. Concentration of C-reactive protein (CRP) was also determined by a particle...

  2. Proteomic Analysis of Bovine Pregnancy-specific Serum Proteins by 2D Fluorescence Difference Gel Electrophoresis

    Directory of Open Access Journals (Sweden)

    Jae Eun Lee

    2015-06-01

    Full Text Available Two dimensional-fluorescence difference gel electrophoresis (2D DIGE is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. The purpose of this study was to investigate bovine pregnancy-specific proteins in the proteome between bovine pregnant and non-pregnant serum using DIGE technique. Serums of 2 pregnant Holstein dairy cattle at day 21 after artificial insemination and those of 2 non-pregnant were used in this study. The pre-electrophoretic labeling of pregnant and non-pregnant serum proteins were mixed with Cy3 and Cy5 fluorescent dyes, respectively, and an internal standard was labeled with Cy2. Labeled proteins with Cy2, Cy3, and Cy5 were separated together in a single gel, and then were detected by fluorescence image analyzer. The 2D DIGE method using fluorescence CyDye DIGE flour had higher sensitivity than conventional 2D gel electrophoresis, and showed reproducible results. Approximately 1,500 protein spots were detected by 2D DIGE. Several proteins showed a more than 1.5-fold up and down regulation between non-pregnant and pregnant serum proteins. The differentially expressed proteins were identified by MALDI-TOF mass spectrometer. A total 16 protein spots were detected to regulate differentially in the pregnant serum, among which 7 spots were up-regulated proteins such as conglutinin precursor, modified bovine fibrinogen and IgG1, and 6 spots were down-regulated proteins such as hemoglobin, complement component 3, bovine fibrinogen and IgG2a three spots were not identified. The identified proteins demonstrate that early pregnant bovine serum may have several pregnancy-specific proteins, and these could be a valuable information for the development of pregnancy-diagnostic markers in early pregnancy bovine serum.

  3. Human serum albumin-coated gold nanoparticles for selective extraction of lysozyme from real-world samples prior to capillary electrophoresis.

    Science.gov (United States)

    Yeh, Pei-Rong; Tseng, Wei-Lung

    2012-12-14

    This study describes the use of human serum albumin (HSA)-modified gold nanoparticles (HSA-AuNPs) for the selective extraction and enrichment of high-pI protein, lysozyme (Lyz) prior to analysis by capillary electrophoresis (CE) with UV detection. HSA-AuNPs are capable of extracting Lyz from a complex matrix because a HSA capping layer not only stabilizes gold nanoparticles in a high-salt environment but also exhibits strong electrostatic attraction with Lyz under neutral pH condition. Efficient separation of Lyz and other high-pI proteins has been successfully achieved by the filling of cationic polyelectrolyte, poly(diallydimethylammonium chloride) (PDDAC), to the background electrolyte. After capturing Lyz with HSA-AuNPs, PDDAC-filled CE can be directly used for the analysis of the extracted Lyz without the addition of the releasing agent into the extractor. The extraction efficiency relied on the pH of the solution and the concentration of HSA-AuNPs. Under optimal extraction conditions, the limit of detection at a signal-to-noise ratio of 3 for Lyz was down to 8 nM. The combination of HSA-AuNP extraction and PDDAC-filled CE has been applied the analyses of Lyz in hen egg white, human milk, and human tear. Also, this NP-based extraction can be coupled to matrix-assisted desorption/ionization time-of-flight mass spectrometry and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Copyright © 2012 Elsevier B.V. All rights reserved.

  4. Phylogenetic analysis of Aceh cattle breed of Indonesia through mitochondrial D-Loop region

    Directory of Open Access Journals (Sweden)

    Eka Meutia Sari

    2016-06-01

    Full Text Available The objective of this research was to find the basic data on genetic diversity of mtDNA D-Loop in Aceh cattle and its association with Bhutanese, Chinese, and Indian cattle. There were sixty samples of DNA which had been sequenced; i.e. Banda Aceh (11, Saree (20, and Indrapuri (29. To the best of our knowledge this is the first published data on the complete mitochondrial D-Loop sequence of Aceh cattle. Results show that Aceh cattle have the closest relationship to Bos indicus and have been influenced by Bos taurus. The closest genetic ranges among Aceh cattle, Bhutanese, Chinese, Indian and Zebu were Aceh–Zebu (0.0138, Aceh–Bhutanese (0.0156, Aceh–Chinese (0.0190 and Aceh–Indian (0.0193. D-Loop mtDNA analyses showed that there were 27 haplotypes in which twenty-one samples spread in haplotype 1, two samples were in haplotype 2, and the other four haplotypes had various samples in the range of three to seventeen samples. One sample of Aceh cattle from Saree has a closest maternal genetic with B. taurus. One of the four mutations among the star-shaped clusters on median joining network was a new specific haploid-group in Aceh cattle. From this finding it could be assumed that Aceh cattle form a specific haplotype and it can be conclude that Aceh cattle are animal genetic resources from Aceh in Sumatera Island that have to be preserved.

  5. Combination of atomic force microscopy and mass spectrometry for the detection of target protein in the serum samples of children with autism spectrum disorders

    Science.gov (United States)

    Kaysheva, A. L.; Pleshakova, T. O.; Kopylov, A. T.; Shumov, I. D.; Iourov, I. Y.; Vorsanova, S. G.; Yurov, Y. B.; Ziborov, V. S.; Archakov, A. I.; Ivanov, Y. D.

    2017-10-01

    Possibility of detection of target proteins associated with development of autistic disorders in children with use of combined atomic force microscopy and mass spectrometry (AFM/MS) method is demonstrated. The proposed method is based on the combination of affine enrichment of proteins from biological samples and visualization of these proteins by AFM and MS analysis with quantitative detection of target proteins.

  6. Immune response in cattle vaccinated against rabies

    Directory of Open Access Journals (Sweden)

    Oliveira Alexandre Nunes de

    2000-01-01

    Full Text Available In order to determine the best type of rabies vaccine to use as a booster, 78 serological samples from singly vaccinated cattle were analyzed by counterimmunoelectrophoresis technique. The animals were divided into several groups, received the first vaccine dose with modified live virus vaccine (ERA strain and were revaccinated with inactivated virus or modified live virus vaccines. Boosters were given at 2, 4, 8, 12 and 16 weeks following first vaccination. Results showed high titres in the cases of booster with inactivated vaccine. In all cases, however, detectable antibody titres declined quickly.

  7. Economcis of cattle fattening with crop residues in Northern Guinea ...

    African J