Eimeria maxima: The influence of host genotype on parasite reproduction as revealed by quantitative real-time PCR

Blake, Damer P.; Hesketh, P; Archer, Andrew; Shirley, M. W.; Smith, Adrian L.

doi:10.1016/j.ijpara.2005.09.011

Cited by 43 publications

(51 citation statements)

References 64 publications

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“…Knowledge of Eimeria species at the genomic level is continuously emerging, and objective molecular methods for Eimeria species differentiation have been developed (Stucki et al, 1993;Tsuji et al, 1997;Schnitzler et al, 1998Schnitzler et al, , 1999Gasser et al, 2001Gasser et al, , 2005Su et al, 2003;Lien et al, 2007;Haug et al, 2007). Nevertheless, the practical implementation of these techniques in routine diagnostics and epidemiological studies of chicken coccidiosis have so far been limited (Lew et al, 2003;Gasser et al, 2005;Blake et al, 2006;Morris et al, 2007a,b).…”

Section: Introductionmentioning

confidence: 99%

Coccidial infections in commercial broilers: epidemiological aspects and comparison ofEimeriaspecies identification by morphometric and polymerase chain reaction techniques

et al. 2008

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The objective of this study was to add to existing knowledge of the epidemiology and the aetiology of coccidial infections in commercial broiler flocks. Polymerase chain reaction (PCR) and morphometric identification of the Eimeria species were compared as means of differentiation in the field samples of faeces and litter. For morphometry, the Eimeria species were categorized into three groups based on lengths of the oocysts. Two random samples of commercial broilers were studied, one during 2000/01 and the other during 2003/04. The prophylactic regime (in-feed narasin), husbandry and methods applied were broadly the same for both subpopulations. Coccidial infection prevalence increased from approximately 45% to approximately 75% during this period, but infection levels (oocysts per gram of faeces) did not significantly change. There were substantial geographical differences in both prevalence and infection levels. A change in Eimeria species profile occurred during the study period. Five Eimeria species were identified at slaughter, by PCR targeting the ITS-1 region of the genome; Eimeria acervulina (100%), Eimeria tenella (77%), Eimeria maxima (25%), Eimeria praecox (10%) and Eimeria necatrix (2%). PCR and morphometric tentative identification were in complete agreement in only 49% of the cases.

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Section: Introductionmentioning

confidence: 99%

Coccidial infections in commercial broilers: epidemiological aspects and comparison ofEimeriaspecies identification by morphometric and polymerase chain reaction techniques

et al. 2008

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“…The ability to extract eimerian DNA of a quality suitable for LAMP using equipment no more specialised than a microcentrifuge and a water bath, supplemented by inhibitor adsorption using chelex resin, now promotes the wider use of molecular biology in eimerian diagnostics. Intriguingly, the reported detection of quantitative PCR-measurable Eimeria DNA in intestinal tissue 20 days after the initiation of parasite infection, 11 days after the last detectable oocyst output, raises the suggestion that LAMP may be used to detect resolved parasite exposure as well as ongoing infection, even after any visible lesions may have been resolved 25 …”

Section: Discussionmentioning

confidence: 99%

Loop-mediated Isothermal Amplification (LAMP) Assays for the Species-specific Detection of <em>Eimeria</em> that Infect Chickens

Barkway

Pocock

Vrba

et al. 2015

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Eimeria species parasites, protozoa which cause the enteric disease coccidiosis, pose a serious threat to the production and welfare of chickens. In the absence of effective control clinical coccidiosis can be devastating. Resistance to the chemoprophylactics frequently used to control Eimeria is common and sub-clinical infection is widespread, influencing feed conversion ratios and susceptibility to other pathogens such as Clostridium perfringens. Despite the availability of polymerase chain reaction (PCR)-based tools, diagnosis of Eimeria infection still relies almost entirely on traditional approaches such as lesion scoring and oocyst morphology, but neither is straightforward. Limitations of the existing molecular tools include the requirement for specialist equipment and difficulties accessing DNA as template. In response a simple field DNA preparation protocol and a panel of species-specific loop-mediated isothermal amplification (LAMP) assays have been developed for the seven Eimeria recognised to infect the chicken. We now provide a detailed protocol describing the preparation of genomic DNA from intestinal tissue collected post-mortem, followed by setup and readout of the LAMP assays. Eimeria species-specific LAMP can be used to monitor parasite occurrence, assessing the efficacy of a farm's anticoccidial strategy, and to diagnose sub-clinical infection or clinical disease with particular value when expert surveillance is unavailable.

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“…qPCR of the eimerian 5S rDNA has been used for total genome estimation of Eimeria, irrespective of species (Blake et al, 2006), and a variety of additional qPCR assays have been developed for species-specific quantitative identification of different Eimeria species (Blake et al, 2006;Morgan et al, 2009;Vrba et al, 2010). However, the advantage of this method is fully exploited only if truly quantitative estimations (or relative abundance) of each species are obtained (Morgan et al, 2009).…”

Section: Discussionmentioning

confidence: 99%

“…The extracted DNA was subjected to qPCR for Eimeria genome quantification using the 5S rDNA as target (Blake et al, 2006). qPCR was performed in a Realplex 4S cycler (Eppendorf, Hamburg, Germany) using 500 nM primer concentration, 250 nM probe (labelled with FAM and TAMRA) concentration (sequences as described by Blake et al, 2006), and 2 ml undiluted extracted DNA with the TaqMan † Universal PCR Master Mix (Applied Biosystems, Foster City, California, USA). The qPCR was performed in triplicate with the following cycle conditions: 508C for 2 min, 10 min at 958C and 40 cycles of 958C for 15 sec and 608C for 1 min.…”

Section: Methodsmentioning

confidence: 99%

“…The advantages of these molecular methods are their reduced requirement for specialist parasitological knowledge, speed, specificity, discriminatory power, objectivity, high throughput and automation potential. Total genome numbers can be estimated using the multicopy 5S rDNA as target (Blake et al, 2006;Vrba et al, 2010) and oocyst numbers calculated using standard or microscopically determined sporulation percentages (Morgan et al, 2009). One of the disadvantages of these techniques in addition to the costs involved is their susceptibility to the presence of PCR inhibitors (Hartman et al, 2005;Sarfo et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

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Real-time PCR-based quantification ofEimeriagenomes: a method to outweigh underestimation of genome numbers due to PCR inhibition

et al. 2013

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Eimeria maxima: The influence of host genotype on parasite reproduction as revealed by quantitative real-time PCR

Cited by 43 publications

References 64 publications

Coccidial infections in commercial broilers: epidemiological aspects and comparison ofEimeriaspecies identification by morphometric and polymerase chain reaction techniques

Coccidial infections in commercial broilers: epidemiological aspects and comparison ofEimeriaspecies identification by morphometric and polymerase chain reaction techniques

Loop-mediated Isothermal Amplification (LAMP) Assays for the Species-specific Detection of <em>Eimeria</em> that Infect Chickens

Real-time PCR-based quantification ofEimeriagenomes: a method to outweigh underestimation of genome numbers due to PCR inhibition

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