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J J Cerón - One of the best experts on this subject based on the ideXlab platform.
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Diagnostic accuracy of porcine acute phase proteins in Meat Juice for detecting disease at abattoir
Veterinary Record, 2015Co-Authors: A M Gutiérrez, S Martínez-subiela, J J CerónAbstract:The aim of this work was to evaluate whether acute phase protein (APP) determinations could assist Official Veterinarians carrying out work in slaughterhouses. To test this hypothesis, the diagnostic accuracy of APP determinations in Meat Juice of pigs was analysed to differentiate between healthy and diseased pigs. One hundred and one pigs of two different origins were classified into two groups according to their health status (healthy and diseased pigs), which was determined by a veterinary clinical examination on the farm. To assess the pigs’ immune status, against the main porcine diseases, serological analyses were monitored. A general idea of the degree of disease coverage was analysed by examining organ lesions postmortem. Haptoglobin (Hp) and C reactive protein (CRP) were measured in Meat Juice samples. 72.13 per cent of pigs appeared to be seropositive for the porcine respiratory and reproductive syndrome virus, and almost 86.2 per cent of them had concomitant infections with other pathogens, such as Porcine circovirus type 2 or Swine influenza virus. Median Hp and CRP concentrations were significantly higher in diseased animals at different stages of the production chain, when compared with levels found in healthy finishing pigs (P
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Towards a better understanding of salivary and Meat Juice acute phase proteins determination in pigs: An expression study
Veterinary Immunology and Immunopathology, 2013Co-Authors: Laura Soler-vasco, J J Cerón, A. Gutiérrez, A. Müllebner, J.c. DuvigneauAbstract:Acute phase proteins (APPs) determination in different fluids like serum, saliva and Meat Juice measured with ultrasensitive assays can be used to evaluate the disease status of porcine populations under field conditions. Liver is the main production site of serum APPs, but the origin of APPs that can be determined in body fluids different from blood remains unknown. The objective of this study was to clarify the origin of three APPs: C-reactive protein (CRP), serum amyloid A (SAA) and haptoglobin (Hp) in saliva and Meat Juice. The mRNA expression of these proteins was measured in liver, salivary gland and diaphragmatic muscle by quantitative PCR and compared with the protein levels in serum, saliva and Meat Juice, respectively in healthy and naturally diseased animals. As expected, concentrations of all APP were significantly higher in all body fluids from diseased animals. Levels of all APPs mRNA were very low in diaphragmatic muscle tissue, and the expression was independent of the disease status. In contrast, we found higher expression levels of SAA and Hp mRNA in the salivary gland of diseased animals, while CRP mRNA was not detected. Our data indicate that the APP present in Meat Juice derived predominantly from serum. This assumption is also supported by the good correlation of the levels of both proteins in Meat Juice with those in serum. Further, the lower variability of the APP levels within the two groups of animals, suggests Meat Juice as an alternate sampling material. The APP levels that are determined in saliva, however, appear to result from an increased local production except for CRP, indicating that the salivary gland responds to disease. These findings are relevant for the establishment of saliva as the preferred diagnostic sample for health monitoring programmes, due to the technical and ethical advantages of the collection.
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Dual-label time-resolved fluoroimmunoassay for simultaneous quantification of haptoglobin and C-reactive protein in Meat Juice from pigs.
Canadian journal of veterinary research = Revue canadienne de recherche veterinaire, 2012Co-Authors: A M Gutiérrez, M D Parra, J J Cerón, Blas A. Marsilla, S Martínez-subielaAbstract:A new method was developed to simultaneously measure 2 acute-phase proteins (APPs) by time-resolved immunofluorometry. The assay, based on double-label quantification of haptoglobin (Hp) and C-reactive protein (CRP) in Meat Juice samples from pigs, was constructed by use of a combination of europium and samarium chelate lanthanides as labels. Meat Juice samples from 154 pigs were used for analytic and clinical validation of the assay through determination of precision, accuracy, limit of detection, and quantification. The analytic performance of the assay was satisfactory, with good intra-assay and interassay precision and accuracy. The levels of Hp and CRP were increased in the Meat Juice samples of diseased animals compared with healthy ones. According to the results, higher sensitivity could be achieved if the cut-off values of both proteins were taken into account for clinical relevance rather than used individually. Since the dual assay saved both time and sample, it could be used as a rapid and sensitive screening test in porcine production.
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Fast measurement of serum amyloid A in different specimens from swine by using a new one-step time-resolved fluorescent immunoassay:
Journal of Veterinary Diagnostic Investigation, 2011Co-Authors: Laura Soler, A M Gutiérrez, S Martínez-subiela, J J CerónAbstract:The objective of the present study was to develop a one-step, fast, competitive time-resolved fluorescent immunoassay to determine porcine serum amyloid A (SAA) by using species-specific reagents. The assay consisted of an all-in-one format involving only 55 min of incubation that was adapted and validated for use in 3 different specimens: serum, saliva, and Meat Juice. The method had overall within- and between-run coefficients of variation under 8% and 12%, respectively, and coefficients of determination higher than 0.93 for linearity under dilution analysis for all specimens. The limits of detection were 0.32 mg/l, 0.28 mg/l, and 1.74 mg/l for serum, saliva, and Meat Juice measurements, respectively. Upper and lower limits of quantification were determined for each sample type and resulted in wide assay ranges that allowed a precise SAA measurement in all the fluids investigated. Statistically significant differences (P = 0.0004 for serum and P < 0.0001 for the saliva and Meat Juice samples) in SAA lev...
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Evaluation of changes in haptoglobin and C-reactive protein concentrations caused by freezing of saliva and Meat Juice samples collected from healthy and diseased pigs
American Journal of Veterinary Research, 2011Co-Authors: A M Gutiérrez, S Martínez-subiela, J J CerónAbstract:Objective—To evaluate changes in stability of haptoglobin and C-reactive protein (CRP) concentrations caused by freezing of saliva and Meat Juice samples. Animals—16 specific-pathogen-free pigs and 16 pigs with clinical signs of disease. Procedures—Saliva and diaphragmatic muscle were collected immediately before and after slaughter, respectively. Haptoglobin and CRP concentrations of pooled samples were measured before storage (day 0) and after 7, 15, 30, 60, 120, 210, and 365 days of storage at −20°C and after repeated freezing-thawing cycles (up to 7 times). In a second experiment, addition of a protease-inhibitor cocktail to saliva and storage of saliva samples at −80°C for up to 30 days were assessed for effects on CRP concentrations. Results—Haptoglobin concentrations in saliva did not change for up to 120 days in samples stored at −20°C, but longer storage times and multiple freezing-thawing cycles increased haptoglobin concentrations. Salivary CRP concentrations decreased significantly after 7 day...
Niels Christian Feld - One of the best experts on this subject based on the ideXlab platform.
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Single versus double testing of Meat-Juice samples for Salmonella antibodies, in the Danish pig-herd surveillance programme.
Preventive Veterinary Medicine, 2003Co-Authors: Lars Ekeroth, Lis Alban, Niels Christian FeldAbstract:Abstract In Denmark, a national serological surveillance-and-control programme for Salmonella in pigs has been in operation since 1995. The programme is based on the Danish mix-ELISA and uses double testing (two ELISA-wells used per sample) of Meat-Juice samples taken in relation to slaughter. All herds are classified monthly into one of the three levels; the classification is based on the percentage of positive serological results in the previous 3 months. In connection with evaluation of the programme in 2001, we investigated whether single testing (testing in one well only) could be expected to be sufficiently precise compared to double testing. Data from the year 2000 were used, and mathematical modelling. Single testing was simulated by randomised selection of one of the two results in the double testing. A slight increase in the prevalence of Salmonella-positive samples (1.02–1.09 times more through the four quarters of the year 2000) was found in the simulated single testing, as compared to the double testing. Around 0.5% of the herds would be allocated to another herd level in single testing—almost equal numbers one level up and one level down. No herd being seronegative in double testing would be allocated to levels 2 or 3 (herds with >40 or >70%, respectively, serological reactors) in single testing. The prevalence of “false-positive” diagnoses (positive in single testing and negative in double testing) and inversely defined “false-negative” diagnoses varied from 4.2 to 8.7% and from 3.2 to 4.5%, respectively, through the four quarters of the year 2000. The probability of allocating a herd to a wrong level due to sampling error was on the average 6.2 (varying from 1.66 to over 100) times higher than the probability of allocating a herd to a wrong level due to the test inaccuracy introduced by going from double to single testing. This is, however, an average; a herd with a true prevalence close to one of the level border cut-offs (40 and 70% weighted seroprevalence, respectively) would have a higher risk of being allocated to a wrong level than a herd with a true prevalence far from the level border cut-offs. The results are based on the current Danish sample sizes in the surveillance scheme, which implies that 60, 75 or 100 samples are taken annually in a herd, depending on its size. Other sample sizes would produce other results.
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Monitoring porcine reproductive and respiratory syndrome virus infection status in swine herds based on analysis of antibodies in Meat Juice samples.
Veterinary Research, 2001Co-Authors: S. Mortensen, Bertel Strandbygaard, Anette Bøtner, Niels Christian Feld, Preben WillebergAbstract:An indirect ELISA test was developed as a novel tool aimed at monitoring the herd infection status of swine herds. Meat Juice samples from pig carcasses were analysed for the presence of antibodies against porcine reproductive and respiratory syndrome virus (PRRSV). A study of samples from herds with known PRRS status was undertaken. The PRRS status of the herds was evaluated based on the analysis of blood samples by another serological test (blocking ELISA) capable of differentiating between infection with PRRSV of the American type and European type. The specificity of the indirect ELISA test on Meat Juice samples was 0.98. The sensitivity of the test depended on the type of the PRRSV strain involved. The apparent prevalence in herds infected with the American type of PRRSV was 0.44. The apparent prevalence in herds infected with the European type of PRRSV was 0.64. Herd level sampling and herd level criteria for assessing the PRRS status of herds by the new test were developed. Herds were classified as PRRS negative or PRRS seropositive based on 10 Meat Juice samples collected randomly at slaughter throughout a 3-month-period. Herd PRRS status classification by the indirect ELISA was validated in 47 herds by collection of blood samples from the herds. Eighteen herds were classified as PRRS negative by both test systems. Twenty-nine herds were classified as PRRS seropositive by both test systems. Acceptable herd classification was achieved using this test.
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nation wide salmonella enterica surveillance and control in danish slaughter swine herds
Preventive Veterinary Medicine, 1997Co-Authors: J Mousing, Niels Christian Feld, J P Nielsen, Thode P Jensen, C Halgaard, F Bager, Berit Jamie Nielsen, S BechnielsenAbstract:Abstract A nation-wide Salmonella enterica surveillance and control programme was initiated in Danish finishing herds over the first quarter of 1995. In Denmark, all swine for slaughter are identifiable by a unique herd code. For each herd code, and depending on the herd's annual kill, random samples ranging from four to more than 60 swine are obtained quarterly at the abattoir. A Meat sample from each pig is frozen, and Meat Juice (harvested after thawing) is examined for specific antibodies against S. enterica using an indirect enzyme-linked immunosorbent assay (ELISA). The ELISA combines several S. enterica O-antigens, and allows detection of antibody response after a variety of different S. enterica serovar infections. Results are transferred to a central database, which each month (based on Meat-Juice tests obtained in the previous 13 weeks) assigns all herds into three S. enterica infection levels: Level 1, in which the S. enterica prevalence is deemed low and acceptable; Level 2, where there is a moderate prevalence of S. enterica seroreacl.ors (from > 50% in the smallest to > 10% in the largest herds); Level 3, in which S. enterica seroreactor prevalence is clearly unsatisfactory ( > 50% for most herd sizes). Irrespective of Salmonella level, all herds receive a monthly update on the current results of the S. enterica test results. If a herd is categorized in Level 2 or 3, it must receive an advisory visit by a practising veterinarian and a local swine extension specialist, and certain management hygiene precautions must be taken. If a herd is categorized in Level 3, the finishers from the herd must additionally be slaughtered under special hygiene precautions. This is supervised by the veterinary authorities. During 1995, 604000 samples were tested for S. enterica, corresponding to 3.0% of the total kill. In December 1995, 15522 herds (representing > 90% of the national production) were categorized into one of the three levels: 14551 herds (93.7%) in Level 1; 610 herds (3.9%) in Level 2; 361 herds (2.3%) in Level 3. The proportion of serologically positive Meat-Juice samples collected during 1995 ranged from a mean of 2.9% in smaller herds (101–200 swine slaughtered per year) to 6.1% in relatively large herds (more than 5000 swine slaughtered per year).
Madalena Vieira-pinto - One of the best experts on this subject based on the ideXlab platform.
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Toxoplasma gondii Infection in Hunted Wild Boars (Sus scrofa): Heart Meat Juice as an Alternative Sample to Serum for the Detection of Antibodies
EcoHealth, 2015Co-Authors: Catarina Coelho, Ana Patrícia Lopes, João Rodrigo Mesquita, Luís Cardoso, Madalena Vieira-pintoAbstract:Toxoplasmosis is a global zoonosis caused by the protozoan parasite Toxoplasma gondii . Detection of antibodies to T . gondii in serum samples from hunted animals may represent a key step for public health protection. It is also important to assess the circulation of this parasite in wild boar population. However, in hunted animals, collection of blood is not feasible and Meat Juice may represent an alternative sample. The purpose of the present study was to evaluate heart Meat Juice of hunted wild boars as an alternative sample for post-mortem detection of antibodies to T . gondii by modified agglutination test (MAT). The agreement beyond chance between results from Meat Juice assessed with Cohen’s kappa coefficient revealed that the 1:20 Meat Juice dilution provided the highest agreement. McNemars’s test further revealed 1:10 as the most suitable Meat Juice dilution, as the proportion of positive paired samples (serum and Meat Juice from the same animal) did not differ at this dilution. All together, these results suggest a reasonable accuracy of heart Meat Juice to detect antibodies to T . gondii by MAT and support it as an alternative sample in post-mortem analysis in hunted wild boars.
S Martínez-subiela - One of the best experts on this subject based on the ideXlab platform.
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Diagnostic accuracy of porcine acute phase proteins in Meat Juice for detecting disease at abattoir
Veterinary Record, 2015Co-Authors: A M Gutiérrez, S Martínez-subiela, J J CerónAbstract:The aim of this work was to evaluate whether acute phase protein (APP) determinations could assist Official Veterinarians carrying out work in slaughterhouses. To test this hypothesis, the diagnostic accuracy of APP determinations in Meat Juice of pigs was analysed to differentiate between healthy and diseased pigs. One hundred and one pigs of two different origins were classified into two groups according to their health status (healthy and diseased pigs), which was determined by a veterinary clinical examination on the farm. To assess the pigs’ immune status, against the main porcine diseases, serological analyses were monitored. A general idea of the degree of disease coverage was analysed by examining organ lesions postmortem. Haptoglobin (Hp) and C reactive protein (CRP) were measured in Meat Juice samples. 72.13 per cent of pigs appeared to be seropositive for the porcine respiratory and reproductive syndrome virus, and almost 86.2 per cent of them had concomitant infections with other pathogens, such as Porcine circovirus type 2 or Swine influenza virus. Median Hp and CRP concentrations were significantly higher in diseased animals at different stages of the production chain, when compared with levels found in healthy finishing pigs (P
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Dual-label time-resolved fluoroimmunoassay for simultaneous quantification of haptoglobin and C-reactive protein in Meat Juice from pigs.
Canadian journal of veterinary research = Revue canadienne de recherche veterinaire, 2012Co-Authors: A M Gutiérrez, M D Parra, J J Cerón, Blas A. Marsilla, S Martínez-subielaAbstract:A new method was developed to simultaneously measure 2 acute-phase proteins (APPs) by time-resolved immunofluorometry. The assay, based on double-label quantification of haptoglobin (Hp) and C-reactive protein (CRP) in Meat Juice samples from pigs, was constructed by use of a combination of europium and samarium chelate lanthanides as labels. Meat Juice samples from 154 pigs were used for analytic and clinical validation of the assay through determination of precision, accuracy, limit of detection, and quantification. The analytic performance of the assay was satisfactory, with good intra-assay and interassay precision and accuracy. The levels of Hp and CRP were increased in the Meat Juice samples of diseased animals compared with healthy ones. According to the results, higher sensitivity could be achieved if the cut-off values of both proteins were taken into account for clinical relevance rather than used individually. Since the dual assay saved both time and sample, it could be used as a rapid and sensitive screening test in porcine production.
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Fast measurement of serum amyloid A in different specimens from swine by using a new one-step time-resolved fluorescent immunoassay:
Journal of Veterinary Diagnostic Investigation, 2011Co-Authors: Laura Soler, A M Gutiérrez, S Martínez-subiela, J J CerónAbstract:The objective of the present study was to develop a one-step, fast, competitive time-resolved fluorescent immunoassay to determine porcine serum amyloid A (SAA) by using species-specific reagents. The assay consisted of an all-in-one format involving only 55 min of incubation that was adapted and validated for use in 3 different specimens: serum, saliva, and Meat Juice. The method had overall within- and between-run coefficients of variation under 8% and 12%, respectively, and coefficients of determination higher than 0.93 for linearity under dilution analysis for all specimens. The limits of detection were 0.32 mg/l, 0.28 mg/l, and 1.74 mg/l for serum, saliva, and Meat Juice measurements, respectively. Upper and lower limits of quantification were determined for each sample type and resulted in wide assay ranges that allowed a precise SAA measurement in all the fluids investigated. Statistically significant differences (P = 0.0004 for serum and P < 0.0001 for the saliva and Meat Juice samples) in SAA lev...
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Evaluation of changes in haptoglobin and C-reactive protein concentrations caused by freezing of saliva and Meat Juice samples collected from healthy and diseased pigs
American Journal of Veterinary Research, 2011Co-Authors: A M Gutiérrez, S Martínez-subiela, J J CerónAbstract:Objective—To evaluate changes in stability of haptoglobin and C-reactive protein (CRP) concentrations caused by freezing of saliva and Meat Juice samples. Animals—16 specific-pathogen-free pigs and 16 pigs with clinical signs of disease. Procedures—Saliva and diaphragmatic muscle were collected immediately before and after slaughter, respectively. Haptoglobin and CRP concentrations of pooled samples were measured before storage (day 0) and after 7, 15, 30, 60, 120, 210, and 365 days of storage at −20°C and after repeated freezing-thawing cycles (up to 7 times). In a second experiment, addition of a protease-inhibitor cocktail to saliva and storage of saliva samples at −80°C for up to 30 days were assessed for effects on CRP concentrations. Results—Haptoglobin concentrations in saliva did not change for up to 120 days in samples stored at −20°C, but longer storage times and multiple freezing-thawing cycles increased haptoglobin concentrations. Salivary CRP concentrations decreased significantly after 7 day...
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Fast measurement of serum amyloid A in different specimens from swine by using a new one-step time-resolved fluorescent immunoassay
Journal of Veterinary Diagnostic Investigation, 2011Co-Authors: Laura Soler-vasco, S Martínez-subiela, Ana Marià Gutiérrez, J J CerónAbstract:The objective of the present study was to develop a one-step, fast, competitive time-resolved fluorescent immunoassay to determine porcine serum amyloid A (SAA) by using species-specific reagents. The assay consisted of an all-in-one format involving only 55 min of incubation that was adapted and validated for use in 3 different specimens: serum, saliva, and Meat Juice. The method had overall within-and between-run coefficients of variation under 8% and 12%, respectively, and coefficients of determination higher than 0.93 for linearity under dilution analysis for all specimens. The limits of detection were 0.32 mg/l, 0.28 mg/l, and 1.74 mg/l for serum, saliva, and Meat Juice measurements, respectively. Upper and lower limits of quantification were determined for each sample type and resulted in wide assay ranges that allowed a precise SAA measurement in all the fluids investigated. Statistically significant differences (P = 0.0004 for serum and P < 0.0001 for the saliva and Meat Juice samples) in SAA levels were found when healthy (n = 20) and diseased (n = 20) pigs were compared. The obtained results indicate that this fast, sensitive, and robust assay for SAA measurement could be of use to determine health and welfare status in swine by employing alternative samples to serum.
David Ranucci - One of the best experts on this subject based on the ideXlab platform.
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Comparison of Serum and Meat Juice for Detection of Anti-Toxoplasma gondii Antibodies in Hunted Wild Boars (Sus scrofa)
Trends in Veterinary Sciences, 2013Co-Authors: David Ranucci, Fabrizia Veronesi, Raffaella Branciari, Dino Miraglia, I. Di Matteo, C. Marini, D. Piergili FiorettiAbstract:Samples of serum and diaphragm Meat Juice obtained from wild boars were tested for anti-Toxoplasma gondii antibodies by means of an indirect immunofluorescent antibody test (IFAT). The results were compared to evaluate the concordance between the two matrices. A total of 160 blood and muscle Juice samples were tested. Antibodies against T. gondii were detected in 28 (17.5 %) blood samples and in 22 Meat Juice samples (13.75 %). The results showed an “almost perfect” concordance between serum and Meat Juice analyses (K value = 0.86). These preliminary data show that diaphragm Meat Juice could be an alternative to serum when using IFAT for T. gondii monitoring purposes in wild boars.
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Evaluation of an Immunofluorescence Antibody Assay for the Detection of Antibodies Against Toxoplasma gondii in Meat Juice Samples from Finishing Pigs
Foodborne Pathogens and Disease, 2012Co-Authors: David Ranucci, Fabrizia Veronesi, Raffaella Branciari, Dino Miraglia, Iolanda Moretta, Daniela Piergili FiorettiAbstract:Serum and Meat Juice analyses for the detection of Toxoplasma gondii antibodies by an immunofluorescence antibody assay were compared in 100 seropositive and 100 seronegative slaughtered heavy swine. Meat Juice was obtained from diaphragm and gracilis muscles of the serologically tested animals. Seventy-two diaphragmatic Meat Juice samples (36%, 95% interval confidence [IC] 29.4%-43.1%) and 63 gracilis Meat Juice samples (31.5%, 95% IC 25.1%-38.4%) tested positive for T. gondii antibodies. The average concordance between serum and Meat Juice derived from both muscles was "substantial" (K=0.6-0.8). The K-value was 1 when considering serum samples showing a titer >1/16, whereas it decreased to 0.62 and to 0.49 when considering serum samples with a 1/16 titer in Meat Juice from diaphragm and gracilis muscle, respectively.