Investigations on the occurrence of tapeworm infections in German horse populations with comparison of different antibody detection methods based on saliva and serum samples

Abstract Background Effective and sustainable worm control in horses would benefit from detailed information about the current regional occurrence of tapeworms. Different diagnostic methods are currently available to detect Anoplocephala spp. infections in horses. However, the format as well as the sensitivity and specificity of the methods vary considerably. Methods A coprological, serological and questionnaire study was conducted to investigate the prevalence and risk factors of tapeworm infections on 48 horse farms in the region of Berlin and Brandenburg, Germany. In total, faecal samples of 484 horses were analysed using the double centrifugation/combined sedimentation-flotation and mini-FLOTAC. Serum (n = 481) and saliva (n = 365) samples were analysed by ELISAs to determine antibody levels against Anoplocephala spp. 12/13 kDa excretory/secretory (E/S) antigens. Results Cestode eggs were detected in 0.6% of faecal samples (farm prevalence 6.3%) without differences between the two methods. In contrast, antibodies against Anoplocephala spp. were detected in 16.2% (farm prevalence 52.1%) and in 29.5% (farm prevalence 75.7%) of the serum and saliva samples, respectively. Both ELISA based methods for detection of tapeworms reported a greater number of infected animals requiring treatment than were positively identified by coproscopy. Logistic regression analysis identified permanent pasture access, large pastures and regular pasture changes and high strongyle egg counts as risk factors for positive serum antibody responses to Anoplocephala spp. while last treatment with praziquantel was protective. Other protective factors were the presence of foals and high numbers of horses on the farm. Daily removal of faeces from the pasture and horse age did not have a significant effect. Conclusions The findings of the present serological investigation indicate that tapeworm prevalence in Berlin/Brandenburg horse farms is much higher than would be anticipated by using conventional/coproscopic analyses. Moreover, the majority of tapeworm-positive horses had not received a cestocidal drug at their last treatment. Considering the already known low sensitivity of the coproscopic detection, the equine veterinary diagnostics can be enhanced by the use of antibody detection methods such as the saliva-based ELISA.

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Rapid Detection of Serum Antibody by Dual-Path Platform VetTB Assay in White-Tailed Deer Infected with Mycobacterium bovis

Clinical and Vaccine Immunology ◽

10.1128/cvi.00120-13 ◽

2013 ◽

Vol 20 (6) ◽

pp. 907-911 ◽

Cited By ~ 18

Author(s):

Konstantin P. Lyashchenko ◽

Rena Greenwald ◽

Javan Esfandiari ◽

Daniel J. O'Brien ◽

Stephen M. Schmitt ◽

...

Keyword(s):

Diagnostic Accuracy ◽

Mycobacterium Bovis ◽

Skin Testing ◽

Serum Antibody ◽

Antibody Responses ◽

Antibody Detection ◽

Serum Samples ◽

Content Type ◽

In The Wild ◽

Free Ranging

ABSTRACTBovine tuberculosis (TB) in cervids remains a significant problem affecting farmed herds and wild populations. Traditional skin testing has serious limitations in certain species, whereas emerging serological assays showed promising diagnostic performance. The recently developed immunochromatographic dual-path platform (DPP) VetTB assay has two antigen bands, T1 (MPB83 protein) and T2 (CFP10/ESAT-6 fusion protein), for antibody detection. We evaluated the diagnostic accuracy of this test by using serum samples collected from groups of white-tailed deer experimentally inoculated withMycobacterium bovis,M. aviumsubsp.paratuberculosis, orM. bovisBCG Pasteur. In addition, we used serum samples from farmed white-tailed deer in herds with no history of TB, as well as from free-ranging white-tailed deer culled during field surveillance studies performed in Michigan known to have bovine TB in the wild deer population. The DPP VetTB assay detected antibody responses in 58.1% of experimentally infected animals within 8 to 16 weeks postinoculation and in 71.9% of naturally infected deer, resulting in an estimated test sensitivity of 65.1% and a specificity of 97.8%. The higher seroreactivity found in deer with naturally acquiredM. bovisinfection was associated with an increased frequency of antibody responses to the ESAT-6 and CFP10 proteins, resulting in a greater contribution of these antigens, in addition to MPB83, to the detection of seropositive animals, compared with experimentalM. bovisinfection. Deer experimentally inoculated with eitherM. aviumsubsp.paratuberculosisorM. bovisBCG Pasteur did not produce cross-reactive antibodies that could be detected by the DPP VetTB assay. The present findings demonstrate the relatively high diagnostic accuracy of the DPP VetTB test for white-tailed deer, especially in the detection of naturally infected animals.

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Cryptosporidiosis in HIV-positive patients and related risk factors: A systematic review and meta-analysis

Parasite ◽

10.1051/parasite/2020025 ◽

2020 ◽

Vol 27 ◽

pp. 27

Author(s):

Ehsan Ahmadpour ◽

Hanie Safarpour ◽

Lihua Xiao ◽

Mehdi Zarean ◽

Kareem Hatam-Nahavandi ◽

...

Keyword(s):

Risk Factors ◽

Data Extraction ◽

Meta Analysis ◽

Diagnostic Methods ◽

Detection Methods ◽

Hiv Positive ◽

Pooled Prevalence ◽

Related Risk ◽

Study Population ◽

Staining Methods

Cryptosporidium is one of the major causes of diarrhea in HIV-positive patients. The aim of this study is to systematically review and meta-analyze the prevalence of Cryptosporidium in these patients. PubMed, Science Direct, Google Scholar, Web of Science, Cochrane and Ovid databases were searched for relevant studies dating from the period of 1 January 2000 to 31 December 2017. Data extraction for the included studies was performed independently by two authors. The overall pooled prevalence was calculated and subgroup analysis was performed on diagnostic methods, geographical distribution and study population. Meta-regression was performed on the year of publication, proportion of patients with diarrhea, and proportion of patients with CD4 < 200 cells/mL. One hundred and sixty-one studies and 51,123 HIV-positive participants were included. The overall pooled prevalence of Cryptosporidium infection in HIV-positive patients was 11.2% (CI95%: 9.4%–13.0%). The pooled prevalence was estimated to be 10.0% (CI95%: 8.4%–11.8%) using staining methods, 13.5% (CI95%: 8.9%–19.8%) using molecular methods, and 26.3% (CI95%: 15.0%–42.0%) using antigen detection methods. The prevalence of Cryptosporidium in HIV patients was significantly associated with the country of study. Also, there were statistical differences between the diarrhea, CD4 < 200 cells/mL, and antiretroviral therapy risk factors with Cryptosporidiosis. Thus, Cryptosporidium is a common infection in HIV-positive patients, and safe water and hand-hygiene should be implemented to prevent cryptosporidiosis occurrence in these patients.

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Detection of Actinobacillus pleuropneumoniae ApxIV toxin antibody in serum and oral fluid specimens from pigs inoculated under experimental conditions

Journal of Veterinary Research ◽

10.1515/jvetres-2017-0021 ◽

2017 ◽

Vol 61 (2) ◽

pp. 163-171 ◽

Cited By ~ 6

Author(s):

Wendy González ◽

Luis G. Giménez-Lirola ◽

Ashley Holmes ◽

Sergio Lizano ◽

Christa Goodell ◽

...

Keyword(s):

Oral Fluid ◽

Serum Antibody ◽

Population Monitoring ◽

Actinobacillus Pleuropneumoniae ◽

Antibody Responses ◽

Antibody Detection ◽

Post Inoculation ◽

Serum Samples ◽

Experimental Conditions ◽

And Control

AbstractIntroduction:The prevention and control ofActinobacillus pleuropneumoniaein commercial production settings is based on serological monitoring. Enzyme-linked immunosorbent assays (ELISAs) have been developed to detect specific antibodies against a variety ofA. pleuropneumoniaeantigens, including long-chain lipopolysaccharides (LPS) and the ApxIV toxin, a repeats-in-toxin (RTX) exotoxin unique toA. pleuropneumoniaeand produced by all serovars. The objective of this study was to describe ApxIV antibody responses in serum and oral fluid of pigs.Material and Methods:Four groups of pigs (six pigs per group) were inoculated withA. pleuropneumoniaeserovars 1, 5, 7, or 12. Weekly serum samples and daily oral fluid samples were collected from individual pigs for 56 days post inoculation (DPI) and tested by LPS and ApxIV ELISAs. The ApxIV ELISA was run in three formats to detect immunlgobulins M, G, and A (IgM, IgG and IgA) while the LPS ELISA detected only IgG.Results:All pigs inoculated withA. pleuropneumoniaeserovars 1 and 7 were LPS ELISA serum antibody positive from DPI 14 to 56. A transient and weak LPS ELISA antibody response was observed in pigs inoculated with serovar 5 and a single antibody positive pig was observed in serovar 12 at ≥35 DPI. Notably, ApxIV serum and oral fluid antibody responses in pig inoculated with serovars 1 and 7 reflected the patterns observed for LPS antibody, albeit with a 14 to 21 day delay.Conclusion:This work suggests that ELISAs based on ApxIV antibody detection in oral fluid samples could be effective in population monitoring forA. pleuropneumoniae.

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Bovine Tuberculosis in a Nebraska Herd of Farmed Elk and Fallow Deer: A Failure of the Tuberculin Skin Test and Opportunities for Serodiagnosis

Veterinary Medicine International ◽

10.4061/2011/953985 ◽

2011 ◽

Vol 2011 ◽

pp. 1-8 ◽

Cited By ~ 27

Author(s):

W. Ray Waters ◽

Gary E. Stevens ◽

Mark A. Schoenbaum ◽

Kathy A. Orloski ◽

Suelee Robbe-Austerman ◽

...

Keyword(s):

Cervus Elaphus ◽

Bovine Tuberculosis ◽

Fallow Deer ◽

Detection Methods ◽

Antibody Detection ◽

Dama Dama ◽

Serologic Test ◽

Serum Samples ◽

Gross Lesions ◽

One Year

In 2009,Mycobacterium bovisinfection was detected in a herd of 60 elk (Cervus elaphus) and 50 fallow deer (Dama dama) in Nebraska, USA. Upon depopulation of the herd, the prevalence of bovine tuberculosis (TB) was estimated at ∼71–75%, based upon histopathology and culture results. Particularly with elk, gross lesions were often severe and extensive. One year ago, the majority of the elk had been tested for TB by single cervical test (SCT), and all were negative. After initial detection of a tuberculous elk in this herd, 42 of the 59 elk were tested by SCT. Of the 42 SCT-tested elk, 28 were TB-infected with only 3/28 reacting upon SCT. After SCT, serum samples were collected from the infected elk and fallow deer from this herd at necropsy and tested by three antibody detection methods including multiantigen print immunoassay, cervidTB STAT-PAK, and dual path platform VetTB (DPP). Serologic test sensitivity ranged from 79 to 97% depending on the test format and host species. Together, these findings demonstrate the opportunities for use of serodiagnosis in the rapid detection of TB in elk and fallow deer.

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BipD of Burkholderia pseudomallei: Structure, Functions, and Detection Methods

Microorganisms ◽

10.3390/microorganisms9040711 ◽

2021 ◽

Vol 9 (4) ◽

pp. 711

Author(s):

Kasturi Selvam ◽

Muhammad Fazli Khalid ◽

Khairul Mohd Fadzli Mustaffa ◽

Azian Harun ◽

Ismail Aziah

Keyword(s):

Sensitivity And Specificity ◽

Burkholderia Pseudomallei ◽

Rapid Determination ◽

Severe Disease ◽

Clinical Manifestations ◽

Culture Method ◽

Laboratory Diagnosis ◽

Detection Methods ◽

Antibody Detection ◽

Low Sensitivity

Melioidosis is a severe disease caused by Burkholderia pseudomallei (B. pseudomallei), a Gram-negative environmental bacterium. It is endemic in Southeast Asia and Northern Australia, but it is underreported in many other countries. The principal routes of entry for B. pseudomallei are skin penetration, inhalation, and ingestion. It mainly affects immunocompromised populations, especially patients with type 2 diabetes mellitus. The laboratory diagnosis of melioidosis is challenging due to its non-specific clinical manifestations, which mimic other severe infections. The culture method is considered an imperfect gold standard for the diagnosis of melioidosis due to its low sensitivity. Antibody detection has low sensitivity and specificity due to the high seropositivity among healthy people in endemic regions. Antigen detection using various proteins has been tested for the rapid determination of B. pseudomallei; however, it presents certain limitations in terms of its sensitivity and specificity. Therefore, this review aims to frame the present knowledge of a potential target known as the Burkholderia invasion protein D (BipD), including future directions for its detection using an aptamer-based sensor (aptasensor).

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Bayesian Estimation of the True Prevalence and of the Diagnostic Test Sensitivity and Specificity of EnteropathogenicYersiniain Finnish Pig Serum Samples

BioMed Research International ◽

10.1155/2015/931542 ◽

2015 ◽

Vol 2015 ◽

pp. 1-7 ◽

Cited By ~ 4

Author(s):

M. J. Vilar ◽

J. Ranta ◽

S. Virtanen ◽

H. Korkeala

Keyword(s):

Bayesian Analysis ◽

Diagnostic Test ◽

Sensitivity And Specificity ◽

Cross Sectional Study ◽

Study Data ◽

Diagnostic Methods ◽

Antibody Detection ◽

Serum Samples ◽

Cross Sectional ◽

True Prevalence

Bayesian analysis was used to estimate the pig’s and herd’s true prevalence of enteropathogenicYersiniain 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 enteropathogenicYersinia. The Bayesian analysis was performed in two steps; the first step estimated the prior true prevalence of enteropathogenicYersiniawith 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 ofYersiniain slaughter-age pigs was 67.5% (95% PI 63.2–70.9). The true prevalence ofYersiniain 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%.

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Performance of Commercial Mycoplasma hyopneumoniae Serum Enzyme-Linked Immunosorbent Assays under Experimental and Field Conditions

Journal of Clinical Microbiology ◽

10.1128/jcm.00485-20 ◽

2020 ◽

Vol 58 (12) ◽

Author(s):

Ana Paula S. Poeta Silva ◽

Ronaldo L. Magtoto ◽

Henrique M. Souza Almeida ◽

Aric McDaniel ◽

Precy D. Magtoto ◽

...

Keyword(s):

Serum Antibody ◽

Mycoplasma Hyopneumoniae ◽

Error Rates ◽

False Positives ◽

Field Conditions ◽

Misclassification Error ◽

Antibody Detection ◽

Serum Samples ◽

Content Type ◽

Immunosorbent Assays

ABSTRACT Mycoplasma hyopneumoniae is an economically significant pathogen of swine. M. hyopneumoniae serum antibody detection via commercial enzyme-linked immunosorbent assays (ELISAs) is widely used for routine surveillance in commercial swine production systems. Samples from two studies were used to evaluate assay performance. In study 1, 6 commercial M. hyopneumoniae ELISAs were compared using serum samples from 8-week-old cesarean-derived, colostrum-deprived (CDCD) pigs allocated to the following 5 inoculation groups of 10 pigs each: (i) negative control, (ii) Mycoplasma flocculare (strain 27399), (iii) Mycoplasma hyorhinis (strain 38983), (iv) Mycoplasma hyosynoviae (strain 34428), and (v) M. hyopneumoniae (strain 232). Weekly serum and daily oral fluid samples were collected through 56 days postinoculation (dpi). The true status of pigs was established by PCR testing on oral fluids samples over the course of the observation period. Analysis of ELISA performance at various cutoffs found that the manufacturers’ recommended cutoffs were diagnostically specific, i.e., produced no false positives, with the exceptions of 2 ELISAs. An analysis based on overall misclassification error rates found that 4 ELISAs performed similarly, although one assay produced more false positives. In study 2, the 3 best-performing ELISAs from study 1 were compared using serum samples generated under field conditions. Ten 8-week-old pigs were intratracheally inoculated with M. hyopneumoniae. Matched serum and tracheal samples (to establish the true pig M. hyopneumoniae status) were collected at 7- to 14-day intervals through 98 dpi. Analyses of sensitivity and specificity showed similar performance among these 3 ELISAs. Overall, this study provides an assessment of the performance of current M. hyopneumoniae ELISAs and an understanding of their use in surveillance.

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Risk Factors for Antimicrobial Resistance in Turkey Farms: A Cross-Sectional Study in Three European Countries

Antibiotics ◽

10.3390/antibiotics10070820 ◽

2021 ◽

Vol 10 (7) ◽

pp. 820

Author(s):

Mayu Horie ◽

Dongsheng Yang ◽

Philip Joosten ◽

Patrick Munk ◽

Katharina Wadepohl ◽

...

Keyword(s):

Risk Factors ◽

Antimicrobial Resistance ◽

Cross Sectional Study ◽

European Countries ◽

Detection Methods ◽

Beta Lactam ◽

Cross Sectional ◽

Shotgun Metagenomics ◽

Faecal Samples ◽

Polymerase Chain

Food-producing animals are an important reservoir and potential source of transmission of antimicrobial resistance (AMR) to humans. However, research on AMR in turkey farms is limited. This study aimed to identify risk factors for AMR in turkey farms in three European countries (Germany, France, and Spain). Between 2014 and 2016, faecal samples, antimicrobial usage (AMU), and biosecurity information were collected from 60 farms. The level of AMR in faecal samples was quantified in three ways: By measuring the abundance of AMR genes through (i) shotgun metagenomics sequencing (n = 60), (ii) quantitative real-time polymerase chain reaction (qPCR) targeting ermB, tetW, sul2, and aph3′-III; (n = 304), and (iii) by identifying the phenotypic prevalence of AMR in Escherichia coli isolates by minimum inhibitory concentrations (MIC) (n = 600). The association between AMU or biosecurity and AMR was explored. Significant positive associations were detected between AMU and both genotypic and phenotypic AMR for specific antimicrobial classes. Beta-lactam and colistin resistance (metagenomics sequencing); ampicillin and ciprofloxacin resistance (MIC) were associated with AMU. However, no robust AMU-AMR association was detected by analyzing qPCR targets. In addition, no evidence was found that lower biosecurity increases AMR abundance. Using multiple complementary AMR detection methods added insights into AMU-AMR associations at turkey farms.

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Evaluation of epizootic situation on arthritis-encephalitis of goats in novosibirsk region

Siberian Herald of Agricultural Science ◽

10.26898/0370-8799-2019-6-12 ◽

2020 ◽

Vol 49 (6) ◽

pp. 104-108

Author(s):

I. S. Onishchenko ◽

I. N. Pen’kova ◽

N. Yu. Balybina ◽

M. A Leonova ◽

V. Yu. Koptev

Keyword(s):

Blood Serum ◽

High Titer ◽

Encephalitis Virus ◽

Diagnostic Methods ◽

Farm Households ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Novosibirsk Region ◽

Private Farm

The analysis of the epizootic situation for viral arthritis-encephalitis of goats in the territory of Novosibirsk region was carried out. No speciﬁ c prophylaxis for this disease has been developed, so the earliest diagnostic methods, as well as the study of its epizootology, are relevant. The studies were conducted in 2019. To study the distribution of goats that are positively responsive to viral arthritis-encephalitis, 198 blood serum samples were taken from goats of various genders, breeds and ages in private farm households and farm enterprises located on the territory of Novosibirsky, Iskitimsky, Ordynsky, Kochenevsky, Moshkovsky and Maslyaninsky districts of Novosibirsk region. In order to study the presence of antibodies to goat arthrit-isencephalitis virus in diagnostic titers, an indirect enzyme-linked immunosorbent assay was used with the antibody detection kit against MVV / CAEV in goat serum (ID Screen® MVV / CAEV Indirect Screening test). Of the 198 animals examined, 86 were found to have diagnostically signiﬁ cant titers of antibodies to the goat arthritis- encephalitis virus, which was 43.4% of the studied population. The result for two goats was uncertain, which amounted to 1%. The remaining animals (55.6%) had no antibodies to goat arthritis-encephalitis virus in their blood serum. The maximum number of positively reacting animals was noted in Novosibirsky district – 66.7%. The Maslyaninsky district was second according to the degree of virus carrying, whereby 47.5% of blood serum samples of goats showed a high titer of antibodies to the goat arthritis-encephalitis virus. The data obtained indicate that at least ﬁ ve districts of the Novosibirsk Region have foci of goat arthritis-encephalitis virus.

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Lateral Flow Assay for the Detection of African Swine Fever Virus Antibodies Using Gold Nanoparticle-Labeled Acid-Treated p72

Frontiers in Chemistry ◽

10.3389/fchem.2021.804981 ◽

2022 ◽

Vol 9 ◽

Author(s):

Wenzhuang Zhu ◽

Kaiwen Meng ◽

Yueping Zhang ◽

Zhigao Bu ◽

Dongming Zhao ◽

...

Keyword(s):

Gold Nanoparticle ◽

African Swine Fever Virus ◽

African Swine Fever ◽

High Sensitivity ◽

Lateral Flow ◽

Fever Virus ◽

Lateral Flow Assay ◽

Diagnostic Methods ◽

Detection Methods ◽

Antibody Detection

African swine fever is a widespread and highly contagious disease in the porcine population, which is caused by African swine fever virus (ASFV). The PCR and ELISA detection methods are the main conventional diagnostic methods for ASFV antigen/antibody detection in the field. However, these methods have limitations of expensive equipment, trained technicians, and time-consuming results. Thus, a rapid, inexpensive, accurate and on-site detection method is urgently needed. Here we describe a double-antigen-sandwich lateral-flow assay based on gold nanoparticle-conjugated ASFV major capsid protein p72, which can detect ASFV antibody in serum samples with high sensitivity and specificity in 10 min and the results can be determined by naked eyes. A lateral flow assay was established by using yeast-expressed and acid-treated ASFV p72 conjugated with gold nanoparticles, which are synthesized by seeding method. A high coincidence (97.8%) of the assay was determined using clinical serum compared to a commercial ELISA kit. In addition, our lateral flow strip can detect as far as 1:10,000 diluted clinically positive serum for demonstration of high sensitivity. In summary, the assay developed here was shown to be rapid, inexpensive, accurate and highly selective. It represents a reliable method for on-site ASFV antibody detection and may help to control the ASFV pandemic.

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