Toxoplasma gondii infections in chickens – performance of various antibody detection techniques in serum and meat juice relative to bioassay and DNA detection methods

Schares, Gereon; Koethe, Martin; Bangoura, Berit; Geuthner, Anne-Catrin; Randau, F.; Ludewig, Martina; Maksimov, Pavlo; Sens, Mareen; Bärwald, Andrea; Conraths, Franz Josef; Villena, Isabelle; Aubert, Dominique; Opsteegh, Marieke; Giessen, Joke van der

doi:10.1016/j.ijpara.2018.03.007

Cited by 32 publications

(67 citation statements)

References 49 publications

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“…Sera were screened by ELISA as previously described, making use of purified antigens that are highly specific towards the three investigated parasites, i.e. TgSAG1 (p30) [33][34][35][36][37][38], NcSRS2 (p38) [39] and the APure-BbELISA antigen [40][41][42][43]. We used an anti-cat conjugate (anti-cat (goat) IgG (H&L) peroxidase (#102-035-003; Dianova, Hamburg, Germany) for all Feliformia species including both hyena species.…”

Section: Laboratory Analyses Elisamentioning

confidence: 99%

Species-specific differences in Toxoplasma gondii, Neospora caninum and Besnoitia besnoiti seroprevalence in Namibian wildlife

et al. 2020

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Background: Knowledge about parasitic infections is crucial information for animal health, particularly of free-ranging species that might come into contact with livestock and humans. Methods:We investigated the seroprevalence of three tissue-cyst-forming apicomplexan parasites (Toxoplasma gondii, Neospora caninum and Besnoitia besnoiti) in 506 individuals of 12 wildlife species in Namibia using in-house enzyme linked immunosorbent assays (indirect ELISAs applying purified antigens) for screening and immunoblots as confirmatory tests. We included six species of the suborder Feliformia, four species of the suborder Caniformia and two species of the suborder Ruminantia. For the two species for which we had most samples and life-history information, i.e. cheetahs (Acinonyx jubatus, n = 250) and leopards (Panthera pardus, n = 58), we investigated T. gondii seroprevalence in relation to age class, sex, sociality (solitary, mother-offspring group, independent sibling group, coalition group) and site (natural habitat vs farmland).Results: All but one carnivore species (bat-eared fox Otocyon megalotis, n = 4) were seropositive to T. gondii, with a seroprevalence ranging from 52.4% (131/250) in cheetahs to 93.2% (55/59) in African lions (Panthera leo). We also detected antibodies to T. gondii in 10.0% (2/20) of blue wildebeest (Connochaetes taurinus). Adult cheetahs and leopards were more likely to be seropositive to T. gondii than subadult conspecifics, whereas seroprevalence did not vary with sex, sociality and site. Furthermore, we measured antibodies to N. caninum in 15.4% (2/13) of brown hyenas (Hyaena brunnea) and 2.6% (1/39) of black-backed jackals (Canis mesomelas). Antibodies to B. besnoiti were detected in 3.4% (2/59) of African lions and 20.0% (4/20) of blue wildebeest. Conclusions:Our results demonstrate that Namibian wildlife species were exposed to apicomplexan parasites at different prevalences, depending on parasite and host species. In addition to serological work, molecular work is also needed to better understand the sylvatic cycle and the clear role of wildlife in the epidemiology of these parasites in southern Africa.

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Section: Laboratory Analyses Elisamentioning

confidence: 99%

Species-specific differences in Toxoplasma gondii, Neospora caninum and Besnoitia besnoiti seroprevalence in Namibian wildlife

et al. 2020

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“…A total of 23 non-infected control chickens and 66 inoculated chickens were used, which were orally inoculated with oocysts or brains of chronically infected mice or by intravenous (i.v.) injection of in vitro-cultivated tachyzoites [22]. Three different T. gondii strains were used: the type II T. gondii strain CZ-Tiger [25]; type II T. gondii ME49 [26]; and type III T. gondii NED [27].…”

Section: Parasite Strains and Experimental Infectionsmentioning

confidence: 99%

“…MC-qPCR was essentially performed as described [29] with some slight modi cations [22]. For the PD-qPCR, tissues were digested [11,30] and the qPCR performed on digests as described [31,32] using primers and a probe targeting the 529 bp repeat of T. gondii [33].…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

“…The results have been used to estimate the burden of infections in chickens or on chicken farms (reviewed by [10]) to assess the potential risk for consumers [15], to identify chickens with viable T. gondii infections [16] or to assess risk factors for infection in this livestock species [9,[17][18][19][20]. Suitable serological techniques for chickens include MAT [12,16,21], IFAT [9,21,22] and ELISA [9,21,22].…”

Section: Introductionmentioning

confidence: 99%

“…To determine the diagnostic characteristics of the TgSAG1 bio -BBMA, we used sera and tissues from experimentally or naturally infected chickens. These had been collected in previous studies [9,22], in which we had determined the true infection status in these chickens using magnetic-capture-(MC-) realtime PCR (qPCR). Likewise, a combination of mouse-bioassay, MC-qPCR and quantitative PCR on acidic pepsin muscle digests (PD-qPCR) had been used.…”

Section: Introductionmentioning

confidence: 99%

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Fluorescent bead-based serological detection of Toxoplasma gondii infection in chickens

Fabian

Hedar

Koethe

et al. 2020

Preprint

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Background: Free-ranging chickens are often infected with Toxoplasma gondii and seroconvert upon infection. This indicates environmental contamination with T. gondii. Methods: Here, we established a bead-based multiplex assay (BBMA) using the Luminex technology for the detection of T. gondii infections in chickens. Recombinant biotinylated T. gondii surface antigen 1 (TgSAG1bio) bound to streptavidin-conjugated magnetic Luminex beads served as antigen. Serum antibodies were detected by a fluorophore-coupled secondary antibody. Beads of differing color codes were conjugated with anti-chicken IgY or chicken serum albumin and served for each sample as an internal positive or negative control, respectively. The assay was validated with sera from experimentally and naturally infected chickens. The results were compared to those from reference methods, including other serological tests, PCRs and bioassay in mice.Results: In experimentally infected chickens, the vast majority (98.5%, n = 65/66) of birds tested seropositive in the BBMA. This included all chickens positive by magnetic-capture PCR (100%, n = 45/45). Most, but not all inoculated and TgSAG1bio-BBMA-positive chickens were also positive in two previously established TgSAG1-ELISAs (TgSAG1-ELISASL, n = 61/65; or TgSAG1-ELISASH, n = 60/65), or positive in an immunofluorescence assay (IFAT, n = 64/65)) and in a modified agglutination test (MAT, n = 61/65). All non-inoculated control animals (n = 28/28, 100%) tested negative. In naturally exposed chickens, the TgSAG1bio-BBMA showed a high sensitivity (98.5%; 95% confidence interval, CI: 90.7–99.9%) and specificity (100%; 95% CI: 85.0–100%) relative to a reference standard established using ELISA, IFAT and MAT. Almost all naturally exposed chickens that were positive in bioassay or by PCR tested positive in the TgSAG1bio-BBMA (93.5%; 95% CI: 77.1–98.9%), while all bioassay- or PCR-negative chickens remained negative (100%; 95% CI: 85.0–100%).Conclusions: The TgSAG1bio-BBMA represents a suitable method for the detection of T. gondii infections in chickens with high sensitivity and specificity, which is comparable or even superior to other tests. Since assays based on this methodology allow for the simultaneous analysis of a single biological sample with respect to multiple analytes, the described assay may represent a component in future multiplex assays for broad serological monitoring of poultry and other farm animals for various pathogens.

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Development of a droplet digital polymerase chain reaction tool for the detection of Toxoplasma gondii in meat samples

et al. 2022

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Toxoplasmosis is a zoonotic disease caused by the protozoan parasite Toxoplasma gondii. Infection in humans has usually been related to the consumption of raw, undercooked or cured meat. The aim of this study was to develop a droplet digital polymerase chain reaction (ddPCR)-based assay for the detection and quantification of T. gondii in meat samples. To optimize the ddPCR, T.gondii reference DNA aliquots at five known concentrations: 8000 cg/µl, 800 cg/µl, 80 cg/µl, 8 cg/µl were used. Moreover, results obtained by ddPCR and quantitative PCR (qPCR) were compared using 80 known samples (40 positive and 40 negative), as well as 171 unknown diaphragm tissue samples collected at slaughterhouses. The ddPCR showed a sensitivity of 97.5% and a specificity of 100%, with a detection limit of 8 genomic copy/µl of T. gondii. A nearly perfect agreement (κ = 0.85) was found between results obtained by ddPCR and qPCR for both positive and negative known samples analysed. On the 171 diaphragm tissue samples from field, 7.6% resulted positive by ddPCR and only 1.2% by qPCR. Therefore, this innovative method could be very useful for the detection of T. gondii in meat samples, aiming to prevent human infections.

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Toxoplasma gondii infections in chickens – performance of various antibody detection techniques in serum and meat juice relative to bioassay and DNA detection methods

Cited by 32 publications

References 49 publications

Species-specific differences in Toxoplasma gondii, Neospora caninum and Besnoitia besnoiti seroprevalence in Namibian wildlife

Species-specific differences in Toxoplasma gondii, Neospora caninum and Besnoitia besnoiti seroprevalence in Namibian wildlife

Fluorescent bead-based serological detection of Toxoplasma gondii infection in chickens

Development of a droplet digital polymerase chain reaction tool for the detection of Toxoplasma gondii in meat samples

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