Identification of Eimeria mitis and Eimeria praecox in broiler feces using polymerase chain reaction

Meireles, Marcelo Vasconcelos; Roberto, LO; Riera, RF

doi:10.1590/s1516-635x2004000400010

Cited by 7 publications

(10 citation statements)

References 17 publications

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“…Using a commercial kit identical to that used in our study, Meireles et al (2004) needed to purify DNA samples after the extraction procedure using the Prep-A Gene DNA purification system kit (Bio-Rad ® ) in order to obtain cleaner samples; also, at least 50 Eimeria oocysts were required to obtain good results in individualized PCR. These results show that contaminants in samples complicate the procedures of DNA and PCR extraction.…”

Section: Dicussionmentioning

confidence: 99%

“…Another important factor is the ability of amplification of different species using a single annealing temperature, thereby allowing analysis of a large number of samples at once, as well as permitting the use of multiplex PCR as also found by Fernandez et al (2003b). Other primers for detection of Eimeria species are known, but they require different annealing temperatures, requiring that reactions be performed separately, requiring more time and reagents (Meireles et al, 2004;Prado, 2005;Haug et al, 2007).…”

Section: Dicussionmentioning

confidence: 99%

“…For successful identification, it is necessary to have an efficient procedure for DNA extraction from oocysts; protocols with phenol/chloroform and commercial extraction kits are commonly used to break the rigid oocyst wall (Fernandez et al, 2003b;Meireles et al, 2004;Haug et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

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Molecular diagnosis of Eimeria species affecting naturally infected Gallus gallus

Fs¹,

Aa²,

Teixeira³

et al. 2011

Genet. Mol. Res.

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ABSTRACT. We used PCR to test various protocols and define a technique for DNA extraction directly from chicken-shed stool samples for the identification of Eimeria species that parasitize birds. It was possible to extract and amplify DNA of seven Eimeria species from field stool samples, using both protocols tested; extractions made with phenol/chloroform protocols gave the best results. The primers were specific and sensitive, allowing amplification of samples containing as few as 20 oocysts, both in individual and in a multiplex PCR. Individualized PCR with the phenol/chloroform DNA extraction protocol detected a larger number of Eimeria species. Molecular diagnosis was found to be practical and precise, and can be used for monitoring and epidemiological studies of Eimeria.

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Section: Dicussionmentioning

confidence: 99%

Section: Dicussionmentioning

confidence: 99%

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Molecular diagnosis of Eimeria species affecting naturally infected Gallus gallus

Fs¹,

Aa²,

Teixeira³

et al. 2011

Genet. Mol. Res.

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show abstract

“…This was possibly due to the small number of oocysts in the intestinal content since the praecox showed a frequency of 3.1% and 0%, respectively. As a macroscopic diagnosis is not possible, it is essential to use molecular techniques to verify the presence of these species (Meireles et al 2004, Carvalho et al 2011).…”

Section: Discussionmentioning

confidence: 99%

Comparison of macroscopy, histopathology and PCR for diagnosing Eimeria spp. in broiler chickens

Balestrin

Santiani

et al. 2022

Pesq. Vet. Bras.

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Coccidiosis is a disease of great importance in industrial poultry. The correct diagnosis directs the poultry industry to its best treatment and control. Thus, a survey of Eimeria spp. was carried out in intestines of 64 broiler flocks, with an average age of 29 days. Eight broilers from each flock were randomly removed from the slaughter line, in a total of 512 samples. Macroscopic and histopathological lesions in the intestine were classified into Scores 0 to 4. Polymerase chain reaction (PCR) was used to research the oocysts from the seven species of Eimeria spp. in the intestinal content. The macroscopic evaluations showed that 59.4% (38/64) of the flocks were positive for E. acervulina, 32.8% (21/64) for E. maxima, 29.7% (19/64) for E. tenella, and 34.4% (22/64) for E. brunetti. The histopathological evaluation showed that 87.5% (56/64) of the flocks had at least one broiler with parasitic structures compatible with Eimeria spp. in the duodenum, 70.3% (45/64) in the jejunum, 18.8% (12/64) in the ileum, 46.9% (30/64) in the cecum, and 4.7% (3/64) in the colon. In PCR, 21.9% (14/64) of the flocks were positive for E. acervulina, 12.5% (8/64) for E. maxima, 3.1% (2/64) for E. mitis, and 32.8% (21/64) for E. tenella. The Kappa Cohen test between macroscopy, histopathology, and PCR demonstrated concordance ranging from weak to moderate with the exception of histopathology and PCR of the cecum, which was strong. In the comparison between macroscopy and histopathology, there were significative differences between Scores 0 and 1 (apart from the cecum). For Score 3, there were significative differences in duodenum, jejunum and cecum (p<0.05). In conclusion, the macroscopic diagnosis and PCR can generate false-negative results, and the histopathological exam proved to be effective, making it essential to associate different techniques for the correct diagnosis of Eimeria spp. in broiler chickens.

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“…Conventionally, traditional methods for Eimeria species identification are based on morphology and/or morphometry of the sporulated oocyst, pre‐patent period, sporulation time, and site of pathological lesions due to Eimeria infection (Aarthi et al, 2010; Hadipour et al, 2011). Identification through these methods sometimes lacks accuracy due to overlapping characteristics and mixed infections among different Eimeria species, which leads to a problem for precise Eimeria species discrimination (Carvalho et al, 2011; Meireles et al, 2004; Morgan et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Identification of Eimeria acervulina (Apicomplexa: Eimeriidae) infecting the broiler chicken Gallus gallus domesticus through morphology and molecular analyses

Mohammed,

Al‐Shaebi,

Al‐Quraishy

et al. 2023

Animal Physiology Nutrition

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Coccidiosis is an intestinal protozoan disease that affects the poultry industry worldwide. The severity of this disease varies depending on the identity of the infectious agents. Therefore, this study was carried out to identify the Eimeria species that affect broiler chickens, Gallus gallus domesticus, through morphological and molecular phylogenetic analyses. Twenty‐five faecal samples were collected from the broiler chickens in a commercial poultry farm in Riyadh (Saudi Arabia). Using the floatation technique, faeces were examined microscopically for the Eimeria occurrence. Identification of Eimeria species was performed based on morphological criteria and molecular tools (DNA amplification for the partial small subunit ribosomal RNA (18S rRNA), internal transcribed spacer (ITS)‐1, and mitochondrial cytochrome c oxidase I (COI) genes. In this study, 32% (8 out of 25) of collected samples were found to be positive for coccidiosis. After sporulation in potassium dichromate (K2Cr2O7), the sporulated oocysts were observed as ovoid and measured 18.37–23.19 µm (19.87) long and 15.07–18.67 µm (16.46) wide, with the anterior location of a polar granule and absence of micropyle. These Eimeria oocysts were assumed to size and shape characteristics of Eimeria acervulina. Molecular analysis was conducted on the sequences of the polymerase chain reaction products from the three genes studied (18S rRNA, ITS‐1, and COI). At the three genes, results showed that the resultant sequences clustered with E. acervulina from different regions confirming morphological description. This study highlighted the importance of molecular techniques to detect avian Eimeria species more than the traditional morphology‐based tool to optimise the appropriate anticoccidial strategies for long‐term control in the studied area.

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Identification of Eimeria mitis and Eimeria praecox in broiler feces using polymerase chain reaction

Cited by 7 publications

References 17 publications

Molecular diagnosis of Eimeria species affecting naturally infected Gallus gallus

Molecular diagnosis of Eimeria species affecting naturally infected Gallus gallus

Comparison of macroscopy, histopathology and PCR for diagnosing Eimeria spp. in broiler chickens

Identification of Eimeria acervulina (Apicomplexa: Eimeriidae) infecting the broiler chicken Gallus gallus domesticus through morphology and molecular analyses

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