Comparison of Serological and Molecular Tests for Diagnosis of Caprine Arthritis Encephalitis and Clinical Evaluation of Mammary Glands of Infected Dairy Goats

Background: Caprine Arthritis Encephalitis (CAE) is a disease that causes productive losses in dairy goat flocks due to the reduction in milk production, followed by lesions in joints and mammary glands. An early diagnosis is essential, considering that there is frequent occurrence of asymptomatic animals. Hence, this study aimed to perform a comparison of immunological and molecular based diagnostic tests, represented by Agar Gel Immunodiffusion (AGID), Western Blot (WB) and nested Polymerase Chain Reaction (nPCR). In addition, the mammary glands (MG) of dairy goats were clinically evaluated. Material, Methods & Results: Blood collection and clinical examination were performed in 1191 dairy goats of 12 farms located in Northeastern and Southeastern regions of Brazil. Serological (AGID, WB) and molecular (nPCR) test results were compared and the data, along with MG alterations, were analyzed using Epi-info 7 and WinEpiscope 2.0. Seroprevalence in AGID test was 41.14% (490/1191). In WB, 51.47% (613/1191) of animals were seropositive and nPCR detected 69.44% (827/1191) positive animals. Hence, WB was more sensitive (P < 0.001) than AGID. However, nPCR detected more positive animals than AGID (P < 0.001) and WB (P < 0.001). The analysis of mammary glands revealed that 105 out of 1096 nanny goats presented alterations, of which 101 presented altered consistency, 16 presented elevated temperatures and 60 had enlarged retromammary lymph nodes. There was significant statistic difference (P < 0.05) only when comparing the results of serological tests with MG alterations.Discussion: In general, AGID technique is most frequently used when screening flocks for the disease due to the practicality and low cost this test presents. However, the results demonstrated that AGID detected the lowest number of positive animals. This low sensitivity that the test presented may be attributed to its antigen-antibody interaction mechanism, considering that agar gel precipitation requires multiple interactions. In addition, WB was more effective than AGID in detecting antibodies. On the other hand, nPCR was important for the detection of infected animals that serological tests failed to detect. The intermittence of immunological response observed in the serological tests may be explained by the variation of antibodies levels that may occur during life. Likewise, viral compartmentalization would justify the intermittent detection of proviral DNA. Hence, the results can be influenced by the viral intermittence, test sensitivity, late seroconversion and statistic values that can be calculated (sensitivity, specificity, positive predictive value, negative predictive level and kappa). Crossing the results of the diagnostic tests with the different mammary gland alterations, it was shown that there was a statistically significant difference (P <0.05) only in the comparison of the results of the serological tests with GM alterations. Everything indicates that the humoral or cellular immune system being on stimulus is more propitious to find these changes. In conclusion, WB was more sensitive than AGID and, considering that nPCR can detect a larger number of animals infected with the goat lentivirus, it must be associated with a sensible serological test, such as Western Blot. In addition, infected animals have alterations in MG, which is more frequent in cases with positive serological results.

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Johne’s disease: a successful eradication programme in a dairy goat herd

Veterinary Record ◽

10.1136/vr.104507 ◽

2018 ◽

Vol 182 (17) ◽

pp. 483-483 ◽

Cited By ~ 2

Author(s):

William G Gavin ◽

Catherine A Porter ◽

Nathan Hawkins ◽

Michael J Schofield ◽

John M Pollock

Keyword(s):

Johne's Disease ◽

Johne’S Disease ◽

Dairy Goat ◽

Dairy Goats ◽

Eradication Programme ◽

Agar Gel ◽

Serological Screening ◽

Surveillance Programme ◽

Oral Transmission ◽

Faecal Culture

This retrospective analysis and report describes the successful eradication and posteradication surveillance programme for Johne’s disease (Mycobacterium avium subspecies paratuberculosis (MAP)) in a closed herd of dairy goats. In 1994, MAP’s presence in the goat herd was first suspected through individual annual serological screening and then subsequently confirmed through faecal culture and histopathology in 1997 when implementation of a more aggressive programme of testing and eradication of the diseased animals began. This programme included frequent serological screening of all adult goats using ELISA and agar gel immunodiffusion assays. Faecal cultures for bacteria were performed on suspect or positive animals and for all goats found dead or euthanased, and tissues were submitted for histopathology and acid-fast staining. Additional disease eradication measures included maintaining a closed herd and minimising faecal-oral transmission of MAP. Following a more aggressive testing regimen and euthanasia of goats with positive faecal culture, the herd was first considered free of MAP in 2003 and has remained free to the present day.

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Comparison of Diagnostic Tests for the Detection of Equine Infectious Anemia Antibody

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063878900100114 ◽

1989 ◽

Vol 1 (1) ◽

pp. 50-52 ◽

Cited By ~ 10

Author(s):

Tatsuo Matsushita ◽

Lyndal K. Hesterberg ◽

James P. Porter ◽

Barbara J. Smith ◽

Louis E. Newman

Keyword(s):

Western Blot ◽

Diagnostic Tests ◽

Equine Infectious Anemia Virus ◽

Competitive Elisa ◽

Enzyme Linked Immunosorbent Assay ◽

Veterinary Services ◽

Agar Gel ◽

Equine Infectious Anemia ◽

Low Levels ◽

Anemia Virus

Two diagnostic tests are approved for detecting antibody to equine infectious anemia virus: the agar-gel immunodiffusion (AGID) test and the competitive enzyme-linked immunosorbent assay (ELISA). A total of 420 sera from National Veterinary Services Laboratories check sets were tested with the AGID and competitive ELISA. A 100% correlation was obtained. The AGID and competitive ELISA were further used to test difficult samples with low levels of equine infectious anemia antibody (weak positives). A third test (Western blot) was also used with these weak positive samples to resolve any discordant results.

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Prevalence of Toxoplasma gondii infection in the dairy goat population from an organic farm

Medycyna Weterynaryjna ◽

10.21521/mw.5797 ◽

2017 ◽

Vol 73 (11) ◽

pp. 736-738

Author(s):

Dawid Jańczak ◽

Marcin Świątek ◽

Żaneta Szymańska ◽

Roman Niżnikowski ◽

Elżbieta Gołąb

Keyword(s):

Toxoplasma Gondii ◽

Public Health Problem ◽

Farm Animals ◽

Dairy Goat ◽

Economic Losses ◽

Dairy Goats ◽

Serological Tests ◽

Organic Farm ◽

Goat Population ◽

Toxoplasma Gondii Infection

Protozoal infection of T. gondii is a public health problem and also causes serious economic losses in livestock production in many countries. Farm animals from organic farms are more likely to be infected. The aim of the study was to determine the prevalence of Toxoplasma gondii among 50 dairy goats from an organic farm in the northwestern Poland region and to assess the prevalence of parasite DNA in the milk of infected animals. Serological tests performed by direct agglutination of IgG antibodies against T. gondii were positive in 10% of the tested animals. No parasite DNA was detected in the milk from the seropositive goats. However, the number of tested animals was too small to draw significant epidemiological conclusions.

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Effects of Diluent pH on Enrichment and Performance of Dairy Goat X/Y Sperm

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.747722 ◽

2021 ◽

Vol 9 ◽

Author(s):

Qifu He ◽

Shenghui Wu ◽

Ming Huang ◽

Ying Wang ◽

Kang Zhang ◽

...

Keyword(s):

Ion Concentration ◽

Dairy Goat ◽

Control Group ◽

Sperm Cells ◽

Dairy Goats ◽

Ph Change ◽

Hydrogen Ion Concentration ◽

Expression Of Genes ◽

Significant Difference

In this paper, on the basis of the differences in the hydrogen ion concentration (pH) of the diluent dairy goat semen on X/Y sperm motility, an X/Y sperm enrichment study was conducted to establish a simple and effective method for gender control in dairy goats. Dairy goat semen was diluted using different pH dilutions and was incubated. Then, the X/Y sperm ratio in the isolated upper sperm was determined using the double TaqMan qPCR method. The internal pH change pattern of sperm cells at different pH dilutions was measured using BCECF-AM probe, and the functional parameters of the isolated sperm were tested with the corresponding kit. Next, an in vitro fertilization test was conducted using isolated spermatozoa and oocytes to determine their fertilization rates, the percentages of female embryos, and the expression of genes related to developing potentially fertilized embryos. Results showed that the percentages of the X sperm cells in the upper sperm layer were 67.24% ± 2.61% at sperm dilution pH of 6.2 and 30.45% ± 1.03% at sperm dilution pH of 7.4, which was significantly different from 52.35% ± 1.72% of the control group (pH 6.8) (P < 0.01). Results also showed that there is a relationship between the external pHo and internal pHi of sperm cells. Furthermore, the percentages of female embryos after the in vitro fertilization of the isolated upper sperm with mature oocytes at pH 6.2 and 7.4 were 66.67% ± 0.05 and 29.73% ± 0.04%, respectively, compared with 48.57% ± 0.02% in the control group (pH 6.8). Highly significant differences occurred between groups (P < 0.01). Additionally, no significant difference was observed during the expression of genes related to embryonic development between the blastocysts formed from sperm isolated by changing the pH of the diluent and the control sperm (P > 0.05). Therefore, this study successfully established a simple and effective method for enriched X/Y sperms from dairy goats, which is important for regulating the desired sex progeny during dairy goat breeding and for guiding dairy goat production.

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Human toxocariasis: a seroepidemiological survey in the municipality of Campinas (SP), Brazil

Revista do Instituto de Medicina Tropical de São Paulo ◽

10.1590/s0036-46652002000600002 ◽

2002 ◽

Vol 44 (6) ◽

pp. 303-307 ◽

Cited By ~ 34

Author(s):

Francisco ANARUMA FILHO ◽

Pedro Paulo CHIEFFI ◽

Carlos Roberto Silveira CORREA ◽

Eide Dias CAMARGO ◽

Edilene P. Real da SILVEIRA ◽

...

Keyword(s):

Univariate Analysis ◽

Epidemiological Data ◽

Human Infection ◽

Soil Samples ◽

Blood Collection ◽

Multiple Logistic Regression ◽

Serological Tests ◽

Significant Difference ◽

Santa Monica ◽

Structured Questionnaire

The occurrence of human Toxocara infection was evaluated in three neighborhoods of the periphery of the Campinas municipality (Jardim Santa Mônica, Jardim São Marcos and Jardim Campineiro) in 1999. Forty residences and 138 residents were randomly selected by drawing lots and were submitted to a seroepidemiological survey, which included blood collection for the immunoenzymatic detection (ELISA) of anti-Toxocara antibodies and a blood count, and the application of a semi-structured questionnaire for the evaluation of epidemiological data. Significant levels of anti-Toxocara antibodies were detected in 23.9% of the 1999 samples. No significant difference in the frequency of infection according to age was observed. Environmental contamination with Toxocara eggs was observed in 12.3 and 14.0% of 57 soil samples collected in the same region in December 1998 and July 1999, respectively. Univariate analysis and multiple logistic regression of the data obtained from the questionnaires and of the results of the serological tests, suggest a significant influence of socioeconomic variables on the frequency of human infection with Toxocara under the conditions prevalent in the study area.

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A Specific Immunologic Assay for Functional Plasminogen Activator Inhibitor 1 in Plasma – Standardized Measurements of the Inhibitor and Related Parameters in Patients with Venous Thromboembolic Disease

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646305 ◽

1992 ◽

Vol 68 (05) ◽

pp. 486-494 ◽

Cited By ~ 15

Author(s):

Malou Philips ◽

Anne-Grethe Juul ◽

Johan Selmer ◽

Bent Lind ◽

Sixtus Thorsen

Keyword(s):

Plasminogen Activator ◽

Plasminogen Activator Inhibitor ◽

Normal Subjects ◽

Tissue Type ◽

Blood Collection ◽

Plasminogen Activator Inhibitor 1 ◽

Type Plasminogen Activator ◽

Significant Difference ◽

Activator Inhibitor ◽

Pai 1

SummaryA new assay for functional plasminogen activator inhibitor 1 (PAI-1) in plasma was developed. The assay is based on the quantitative conversion of PAI-1 to urokinase-type plasminogen activator (u-PA)-PAI-l complex the concentration of which is then determined by an ELISA employing monoclonal anti-PAI-1 as catching antibody and monoclonal anti-u-PA as detecting antibody. The assay exhibits high sensitivity, specificity, accuracy, and precision. The level of functional PAI-1, tissue-type plasminogen activator (t-PA) activity and t-PA-PAI-1 complex was measured in normal subjects and in patients with venous thromboembolism in a silent phase. Blood collection procedures and calibration of the respective assays were rigorously standardized. It was found that the patients had a decreased fibrinolytic capacity. This could be ascribed to high plasma levels of PAI-1. The release of t-PA during venous occlusion of an arm for 10 min expressed as the increase in t-PA + t-PA-PAI-1 complex exhibited great variation and no significant difference could be demonstrated between the patients with a thrombotic tendency and the normal subjects.

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Diagnosis of tuberculosis in wildlife: a systematic review

Veterinary Research ◽

10.1186/s13567-020-00881-y ◽

2021 ◽

Vol 52 (1) ◽

Author(s):

Jobin Thomas ◽

Ana Balseiro ◽

Christian Gortázar ◽

María A. Risalde

Keyword(s):

Systematic Review ◽

Test Performance ◽

Diagnostic Tests ◽

Large Scale ◽

Diagnostic Techniques ◽

Review Literature ◽

Meat Industry ◽

Post Mortem ◽

Serological Tests ◽

Ongoing Research

AbstractAnimal tuberculosis (TB) is a multi-host disease caused by members of the Mycobacterium tuberculosis complex (MTC). Due to its impact on economy, sanitary standards of milk and meat industry, public health and conservation, TB control is an actively ongoing research subject. Several wildlife species are involved in the maintenance and transmission of TB, so that new approaches to wildlife TB diagnosis have gained relevance in recent years. Diagnosis is a paramount step for screening, epidemiological investigation, as well as for ensuring the success of control strategies such as vaccination trials. This is the first review that systematically addresses data available for the diagnosis of TB in wildlife following the Preferred Reporting Items of Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The article also gives an overview of the factors related to host, environment, sampling, and diagnostic techniques which can affect test performance. After three screenings, 124 articles were considered for systematic review. Literature indicates that post-mortem examination and culture are useful methods for disease surveillance, but immunological diagnostic tests based on cellular and humoral immune response detection are gaining importance in wildlife TB diagnosis. Among them, serological tests are especially useful in wildlife because they are relatively inexpensive and easy to perform, facilitate large-scale surveillance and can be used both ante- and post-mortem. Currently available studies assessed test performance mostly in cervids, European badgers, wild suids and wild bovids. Research to improve diagnostic tests for wildlife TB diagnosis is still needed in order to reach accurate, rapid and cost-effective diagnostic techniques adequate to a broad range of target species and consistent over space and time to allow proper disease monitoring.

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The Alterations in Methylene Blue/Light-Treated Frozen Plasma Proteins Revealed by Proteomics

Transfusion Medicine and Hemotherapy ◽

10.1159/000515119 ◽

2021 ◽

pp. 1-8

Author(s):

Tiange Wu ◽

Xiaoning Wang ◽

Kai Ren ◽

Xiaochen Huang ◽

Jiankai Liu

Keyword(s):

Mass Spectrometry ◽

Methylene Blue ◽

Western Blot ◽

Blue Light ◽

Differentially Expressed ◽

Enzyme Linked Immunosorbent Assay ◽

Differentially Expressed Protein ◽

Significant Difference ◽

Modified Proteins ◽

Frozen Plasma

Introduction: The aim of this study was to investigate the modified proteins in methylene blue/light-treated frozen plasma (MB-FP) compared with fresh frozen plasma (FFP) in order to gain a better application of MB/light-treated plasma in clinic transfusion. Methods: MB-FP and FFP were collected from Changchun central blood station, and a trichloroacetic acid/acetone precipitation method was used to remove albumin for the enrichment of lower abundance proteins. The plasma protein in MB-FP and FFP were separated using two-dimensional gel electrophoresis (2-DE) and the differentially expressed protein spots were analyzed using mass spectrometry. Finally, the differentially expressed proteins were tested using Western blot and enzyme-linked immunosorbent assay (ELISA). Results: Approximately 14 differentially expressed protein spots were detected in the MB-FP, and FFP was chosen as the control. After 2-DE comparison analysis and mass spectrometry, 8 significantly differentially expressed protein spots were identified, corresponding to 6 different proteins, including complement C1r subcomponent (C1R), inter-alpha-trypsin inhibitor heavy chain H4 (ITI-H4), keratin, type II cytoskeletal 1 (KRT1), hemopexin (HPX), fibrinogen gamma chain (FGG), and transthyretin (TTR). Western blot showed no significant difference in the expression level of KRT1 between MB-FP and FFP (p > 0.05). Both Western blot and ELISA indicated that the level of HPX was significantly higher in FFP than in MB-FP (p < 0.05). Conclusion: This comparative proteomics study revealed that some significantly modified proteins occur in MB-FP, such as C1R, ITI-H4, KRT1, HPX, FGG, and TTR. Our findings provide more theoretical data for using MB-FP in transfusion medicine. However, the relevance of the data for the transfusion of methylene blue/light-treated plasma remains unclear. The exact modification of these proteins and the effects of these modified proteins on their functions and their effects in clinical plasma infusion need to be further studied.

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The effect of clamped and unclamped umbilical cord samples on blood gas analysis

Archives of Gynecology and Obstetrics ◽

10.1007/s00404-021-06076-w ◽

2021 ◽

Author(s):

Elisabetta Colciago ◽

Simona Fumagalli ◽

Elena Ciarmoli ◽

Laura Antolini ◽

Antonella Nespoli ◽

...

Keyword(s):

Blood Gas Analysis ◽

Cross Sectional Study ◽

Gas Analysis ◽

Blood Collection ◽

Large Sample Size ◽

Blood Gas ◽

Delayed Cord Clamping ◽

Cross Sectional ◽

High Risk Women ◽

Significant Difference

Abstract Purpose Delayed cord clamping for at least 60 s is recommended to improve neonatal outcomes. The aim of this study is to evaluate whether there are differences in cord BGA between samples collected after double clamping the cord or without clamping the cord, when blood collection occurs within 60 s from birth in both groups. Methods A cross-sectional study was carried out, collecting data from 6884 high-risk women who were divided into two groups based on the method of cord sampling (clamped vs unclamped). Results There were significant decrease in pH and BE values into unclamped group compared with the clamped group. This difference remained significant when considering pathological blood gas analysis parameters, with a higher percentage of pathological pH or BE values in the unclamped group. Conclusion Samples from the unclamped cord alter the acid–base parameters compared to collection from the clamped cord; however, this difference does not appear to be of clinical relevance. Findings could be due to the large sample size, which allowed to achieve a high power and to investigate very small numerical changes between groups, leading to a statistically significant difference in pH and BE between samples even when we could not appreciate any clinical relevant difference of pH or BE between groups. When blood gas analysis is indicated, the priority should be given to the timing of blood collection to allow reliable results, to assess newborns status at birth and intervene when needed.

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COVID-19 Seroprevalence among Healthcare Workers of a Large COVID-19 Hospital in Rome Reveals Strengths and Limits of Two Different Serological Tests

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph18052650 ◽

2021 ◽

Vol 18 (5) ◽

pp. 2650

Author(s):

Giuseppe Vetrugno ◽

Daniele Ignazio La Milia ◽

Floriana D’Ambrosio ◽

Marcello Di Pumpo ◽

Roberta Pastorino ◽

...

Keyword(s):

Blood Test ◽

Healthcare Workers ◽

Point Of Care ◽

Venous Blood ◽

Administrative Staff ◽

Serological Tests ◽

Predictive Values ◽

Work From Home ◽

Significant Difference ◽

Sensitivity Specificity

Healthcare workers are at the forefront against COVID-19, worldwide. Since Fondazione Policlinico Universitario A. Gemelli (FPG) IRCCS was enlisted as a COVID-19 hospital, the healthcare workers deployed to COVID-19 wards were separated from those with limited/no exposure, whereas the administrative staff were designated to work from home. Between 4 June and 3 July 2020, an investigation was conducted to evaluate the seroprevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) immunoglobulin (IgG) antibodies among the employees of the FPG using point-of-care (POC) and venous blood tests. Sensitivity, specificity, and predictive values were determined with reverse-transcription polymerase chain reaction on nasal/oropharyngeal swabs as the diagnostic gold standard. The participants enrolled amounted to 4777. Seroprevalence was 3.66% using the POC test and 1.19% using the venous blood test, with a significant difference (p < 0.05). The POC test sensitivity and specificity were, respectively, 63.64% (95% confidence interval (CI): 62.20% to 65.04%) and 96.64% (95% CI: 96.05% to 97.13%), while those of the venous blood test were, respectively, 78.79% (95% CI: 77.58% to 79.94%) and 99.36% (95% CI: 99.07% to 99.55%). Among the low-risk populations, the POC test’s predictive values were 58.33% (positive) and 98.23% (negative), whereas those of the venous blood test were 92.86% (positive) and 98.53% (negative). According to our study, these serological tests cannot be a valid alternative to diagnose COVID-19 infection in progress.

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