Sequence diversity of internal transcribed spacer-1 (ITS-1) region of Eimeria infecting chicken and its relevance in species identification from Indian field samples

Bhaskaran, Mohana Subramanian; Venkatesan, Lakshmipriya; Aadimoolam, Ramesh; Jayagopal, Harikrishnan Tirunelveli; Sriraman, Rajan

doi:10.1007/s00436-009-1696-2

Cited by 22 publications

(24 citation statements)

References 24 publications

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“…The present findings are in agreement with the findings of Bhaskaran et al (2009). However involvement of E. tenella in many outbreaks has also been documented from other parts of India (Jithendran 2001;Aarthi et al 2010;Jadhav et al 2011).…”

Section: Discussionsupporting

confidence: 83%

Prevalence of poultry coccidiosis in Jammu region of Jammu & Kashmir State

Sharma

Iqbal²,

Azmi³

et al. 2013

J Parasit Dis

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In this study prevalence of chicken coccidiosis in Jammu division were undertaken in both organized and backyard chickens during the year 2010-2011, with an overall prevalence of 39.58 % on examination of 720 faecal samples. Five Eimeria species were identified viz., E. tenella, E. necatrix, E. maxima, E. acervulina and E. mitis. E. tenella was the predominant species in both organized and unorganized farms. The highest prevalence percentage was found in July, 2011 (68.9 %) and the lowest percentage was found in May, 2011 (12.5 %). Coccidial prevalence was found to be 53.61 % in unorganized (backyard poultry birds) as compared to organized birds (25.55 %). Maximum positive cases of coccidian infection was found in monsoon season (60.55 %) and least in summer season (21.66 %). Birds of age 31-45 days showed more prevalence percentage (58.86 %). Higher oocysts count was recorded from July to September with a peak value (38973.00 ± 3075.6) in July and lowest (12914.00 ± 595.48) in the month of May.

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Section: Discussionsupporting

confidence: 83%

Prevalence of poultry coccidiosis in Jammu region of Jammu & Kashmir State

Sharma

Iqbal²,

Azmi³

et al. 2013

J Parasit Dis

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“…1 and Supplementary Table 2). These gaps may have been due to variations in the ITS-1 sequence, as has been reported previously in the case of E. tenella from India (Bhaskaran et al, 2010). While it is clear that PCR can facilitate the detection of minority Eimeria species sub-populations which may be missed by routine microscopy (Frölich et al, 2013), the reliance of PCR on very small primer annealing sites within a target genome also risks false negatives where genetic diversity occurs.…”

Section: Discussionmentioning

confidence: 59%

An optimised protocol for molecular identification of Eimeria from chickens

Kumar

Garg

Moftah

et al. 2014

Veterinary Parasitology

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Molecular approaches supporting identification of Eimeria parasites infecting chickens have been available for more than 20 years, although they have largely failed to replace traditional measures such as microscopy and pathology. Limitations of microscopy-led diagnostics, including a requirement for specialist parasitological expertise and low sample throughput, are yet to be outweighed by the difficulties associated with accessing genomic DNA from environmental Eimeria samples. A key step towards the use of Eimeria species-specific PCR as a sensitive and reproducible discriminatory tool for use in the field is the production of a standardised protocol that includes sample collection and DNA template preparation, as well as primer selection from the numerous PCR assays now published. Such a protocol will facilitate development of valuable epidemiological datasets which may be easily compared between studies and laboratories. The outcome of an optimisation process undertaken in laboratories in India and the UK is described here, identifying four steps. First, samples were collected into a 2% (w/v) potassium dichromate solution. Second, oocysts were enriched by flotation in saturated saline. Third, genomic DNA was extracted using a QIAamp DNA Stool mini kit protocol including a mechanical homogenisation step. Finally, nested PCR was carried out using previously published primers targeting the internal transcribed spacer region 1 (ITS-1). Alternative methods tested included sample processing in the presence of faecal material, DNA extraction using a traditional phenol/chloroform protocol, the use of SCAR multiplex PCR (one tube and two tube versions) and speciation using the morphometric tool COCCIMORPH for the first time with field samples.

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“…Of the 18 samples, 16 were also found to contain medium sized oocysts (NTP group) while 14 samples were found to contain the large sized oocysts (BM group). Previous studies carried out in Czechoslovakia (Kucera 1990), France (Williams et al 1996) and Korea (Lee et al 2010) reported E. acervulina to be the most highly prevalent species, while studies in China (Sun et al 2009), India (Bhaskaran et al 2010), Jordan (Al-Natour et al 2002 and Iran (Hadipour et al 2013) showed E. tenella to be highly prevalent. These studies conducted in countries from several regions of the world show that both E. acervulina and E. tenella are common among chicken farms, which is consistent with the occurrence of AM and NTP Eimeria species found in the 18 samples.…”

Section: Resultsmentioning

confidence: 94%

Comparison of Molecular Methods for the Detection of Eimeria in Domestic Chickens in Malaysia

Loo

Lim

Efendi

et al. 2019

JSM

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Coccidiosis, caused by the Eimeria species, greatly affects the poultry industry. Severity of the disease varies depending on the identity of the infecting parasites, encouraging identification of Eimeria species circulating on a farm as a valuable component of chicken management. Conventional methods of Eimeria species identification are time consuming and can be subjective in nature. Given these limitations, molecular approaches have been developed for specific detection of Eimeria species. In this study, faecal samples were collected from commercial broiler farms and subjected to microscopic examination for Eimeria occurrence. Eimeria species were putatively identified by morphological characterisation and grouped into three categories based on oocyst size. Molecular detection of Eimeria species occurrence in these samples was then performed using two published PCR assays (the individual components of a SCAR-based multiplex PCR, and assays developed for quantitative PCR, termed PCR-SCAR 1 and PCR-SCAR 2 here) and a LAMP assay. Comparison of the results obtained demonstrated that the three molecular methods were capable of detecting all Eimeria species of the reference Houghton strain, but showed varying efficiencies in detecting Malaysian field isolates. PCR-SCAR 2 was found to be the most effective, detecting all seven Eimeria species and indicating the presence of Eimeria parasites in most flocks. Differences in the ability of the molecular methods to detect Eimeria may be a consequence of sequence divergence between isolates from different regions, implying that development of region-specific methods using local Eimeria strains may be required to improve the efficiency of molecular assays for Eimeria detection.

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Sequence diversity of internal transcribed spacer-1 (ITS-1) region of Eimeria infecting chicken and its relevance in species identification from Indian field samples

Cited by 22 publications

References 24 publications

Prevalence of poultry coccidiosis in Jammu region of Jammu & Kashmir State

Prevalence of poultry coccidiosis in Jammu region of Jammu & Kashmir State

An optimised protocol for molecular identification of Eimeria from chickens

Comparison of Molecular Methods for the Detection of Eimeria in Domestic Chickens in Malaysia

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