scholarly journals DNA extraction methods for molecular detection of Eimeria spp. in cattle and sheep

2020 ◽  
Vol 40 (7) ◽  
pp. 514-518
Author(s):  
Caroline Z. Reginato ◽  
Patrícia Bräunig ◽  
Luiza P. Portella ◽  
Ana Paula G. Mortari ◽  
Camila E. Minuzzi ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Molecular Detection ◽  
Dna Detection ◽  
Eimeria Species ◽  
Extraction Methods ◽  
Ammonium Bromide ◽  
Oocyst Wall ◽  
Fecal Samples ◽  
Eimeria Spp ◽  
Commercial Kit

ABSTRACT: Molecular detection of Eimeria species in fecal samples can be useful for experimental and diagnostic purposes. However, the parasite quantity presence in feces and the oocyst wall are an obstacle in DNA extraction protocols. Therefore, adequate sampling and effective disruption of the oocysts are essential to improve the accuracy of DNA detection by PCR. The aims of this study were to evaluate the suitability of six protocols for DNA extraction from Eimeria spp. present in bovine and sheep. Twenty pools of fecal samples from cattle (10 pools) and sheep (10 pools) were distributed to six DNA extraction protocols: commercial kit, commercial kit with modification, DNAzol, cetyl-trimethyl ammonium bromide (CTAB), glass beads and commercial kit for fecal samples. Fecal samples were submitted to DNA extraction and PCR. Among the protocols tested, CTAB was determined to be most suitable for DNA extraction from oocysts (90% of DNA detection by PCR); DNAzol and CTAB resulted in higher DNA detection from bovine samples (80%). CTAB and commercial kit with modification improved PCR detection of Eimeria spp. in sheep samples, with positive amplification of DNA in all tested samples.

scholarly journals First molecular detection of Eimeria spp. in eared doves (Zenaida auriculata) from Brazil

2020 ◽  
Vol 41 (4) ◽  
pp. 1259
Author(s):  
Alessandra Taroda ◽  
Luiz Daniel de Barros ◽  
Mércia de Seixas ◽  
Sérgio Tosi Cardim ◽  
João Pedro Sasse ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Mitochondrial Genome ◽  
Cytochrome C Oxidase ◽  
Dna Extraction ◽  
Nested Pcr ◽  
Molecular Detection ◽  
Southern Brazil ◽  
Oxidase Gene ◽  
Fecal Samples ◽  
Eimeria Spp

The aim of the present study was to detect Eimeria spp. in eared doves (Zenaida auriculata) from Brazil. Two hundred and fourteen birds were trap-capture in different regions of Londrina city, Paraná state, Southern Brazil. Fecal samples were collected, and DNA extraction was performed. A nested PCR based on the subunit I of the cytochrome c oxidase gene of the Eimeria mitochondrial genome was used to detect the DNA presence of this coccidian in eared dove feces. From 214 birds, 171 (79.9%) were positive for Eimeria spp. There was significantly difference of positivity between the site of capture. When analyzing the genders, the numbers of positive males were 84/103 (81.5%) and females 87/111 (78.4%). To the best of the authors’ knowledge, this is the first study with molecular prevalence of Eimeria sp. in Z. auriculata. Further studies should be done to identify the species of Eimeria that infect eared doves Z. auriculata.

scholarly journals Eimeria infections in goats in Southern Portugal

2014 ◽  
Vol 23 (2) ◽  
pp. 280-286 ◽  
Author(s):  
Liliana Machado Ribeiro da Silva ◽  
Maria João Martins Vila-Viçosa ◽  
Telmo Nunes ◽  
Anja Taubert ◽  
Carlos Hermosilla ◽  
...  
Keyword(s):  
Eimeria Species ◽  
Clinical Signs ◽  
Intestinal Infection ◽  
Dairy Goats ◽  
Diagnostic Investigation ◽  
Highly Pathogenic ◽  
Fecal Samples ◽  
Eimeria Spp ◽  
Southern Portugal ◽  
Prevalent Species

Coccidiosis caused by Eimeria species is a major form of intestinal infection affecting intensively and semi-intensively reared goats. The province of Alentejo is the main goat-producing area in Portugal. Therefore, all 15 Serpentina goat farms in Alentejo were analyzed regarding the occurrence and diversity of Eimeria species. Fecal samples obtained from 144 animals (52.1% dairy goats, 47.9% pre-pubertal goats) were examined using the modified McMaster technique to determine the number of oocysts per gram of feces. Eimeria spp. oocysts were present in 98.61% of the fecal samples and, overall, nine different Eimeria species were identified. The most prevalent species were E. ninakohlyakimovae (88%) and E. arloingi (85%), followed by E. alijevi (63%) and E. caprovina(63%). The average number of oocysts shed was significantly lower in dairy goats than in pre-adult animals. Astonishingly, no clinical signs of coccidiosis were observed in any of the animals examined, even though they were shedding high numbers of oocysts and were infected with highly pathogenic species. Thus, implementation of routine diagnostic investigation of the occurrence and diversity of caprine Eimeria species may be a useful tool for determination and better understanding of their potential economic impact on goat herds in southern Portugal.

Rapid and Effective DNA Extraction Method with Bead Grinding for a Large Amount of Fungal DNA

2010 ◽  
Vol 73 (6) ◽  
pp. 1077-1084 ◽  
Author(s):  
M. WATANABE ◽  
K. LEE ◽  
K. GOTO ◽  
S. KUMAGAI ◽  
Y. SUGITA-KONISHI ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Sodium Dodecyl ◽  
Extraction Methods ◽  
Molecular Techniques ◽  
Ammonium Bromide ◽  
Uv Absorbance ◽  
Chemical Methods ◽  
Wide Range ◽  
Ctab Method ◽  
Commercial Kits

To identify a rapid method for extracting a large amount of DNA from fungi associated with food hygiene, extraction methods were compared using fungal pellets formed rapidly in liquid media. Combinations of physical and chemical methods or commercial kits were evaluated with 3 species of yeast, 10 species of ascomycetous molds, and 4 species of zygomycetous molds. Bead grinding was the physical method, followed by chemical methods involving sodium dodecyl sulfate (SDS), cetyl trimethyl ammonium bromide (CTAB), and benzyl chloride and two commercial kits. Quantity was calculated by UV absorbance at 260 nm, quality was determined by the ratio of UV absorbance at 260 and 280 nm, and gene amplifications and electrophoresis profiles of whole genomes were analyzed. Bead grinding with the SDS method was the most effective for DNA extraction for yeasts and ascomycetous molds, and bead grinding with the CTAB method was most effective with zygomycetous molds. For both groups of molds, bead grinding with the CTAB method was the best approach for DNA extraction. Because this combination also is relatively effective for yeasts, it can be used to extract a large amount of DNA from a wide range of fungi. The DNA extraction methods are useful for developing gene indexes to identify fungi with molecular techniques, such as DNA fingerprinting.

scholarly journals Prevalence and risk factors associated with Eimeria species infection in cattle of different geographical regions of Indonesia

2021 ◽  
pp. 2339-2345
Author(s):  
Fitrine Ekawasti ◽  
Raden Wisnu Nurcahyo ◽  
Lintang Winantya Firdausy ◽  
April Hari Wardhana ◽  
Dyah Haryuningtyas Sawitri ◽  
...  
Keyword(s):  
Risk Factors ◽  
Beef Cattle ◽  
Eimeria Species ◽  
Economic Losses ◽  
Intensive Management ◽  
Adult Cattle ◽  
Fecal Samples ◽  
Geographical Regions ◽  
Eimeria Spp ◽  
Intensively Managed

Background and Aim: Eimeria spp. are gastrointestinal protozoans that affect animal productivity, thereby causing symptoms that range from bloody diarrhea to death. These symptoms cause economic losses to farmers. The distribution of Eimeria spp. in cattle has, therefore, been reported to have spread widely, especially in the tropics and subtropics. Indonesia is a tropical country at high risk of Eimeria infections. This study aimed to identify the prevalence and risk factors related to the levels of eimeriosis in beef cattle originating from different geographic areas in Indonesia. Materials and Methods: Here, 817 fecal samples were collected from beef cattle in Indonesia, including 282 calves, 535 adults, 530 males, and 287 females. In addition, 156 semi-intensively and 661 intensively managed cattle were randomly collected. Then, fecal samples were analyzed by parasitology examinations. Results: Screening examination using the sugar flotation modification method showed that Eimeria spp. were prevalent in Indonesia, as 65.4% of the bacterial strain was detected. The prevalence of identified Eimeria spp. in Indonesia was highest in North Maluku (Maluku Island) (94.1%), whereas the lowest levels were observed in West Java (24.0%) (Java Island). The prevalence was also found to be higher in males (79.3%) than females (51.9%). Similarly, levels in semi-intensively managed cattle (66.7%) were higher than those subjected to intensive management (65.9%). However, its prevalence in calf and adult cattle was similar. Conclusion: Bovine eimeriosis spp. were detected at high prevalence in Indonesia, and high-level risks were observed in infected males, including those under the semi-intensive management. In addition, although the results from oocyst examinations were based on qualitative analysis, the endemicity levels of Eimeria spp. among farms in Indonesia should be considered because Eimeria spp. were distributed in most parts of Indonesia. Based on the results of this study, we provide the first information about the prevalence of bovine eimeriosis from different geographical locations in Indonesia, which have differing climates associated with the level of the existing risk factors. Hence, farmers are advised to pay more attention to strict biosecurity techniques on their farms, thereby favoring the early control of bovine eimeriosis.

The Sensitivity of PCR Detection of Cryptosporidium Oocysts in Fecal Samples Using Two DNA Extraction Methods

2003 ◽  
Vol 7 (3) ◽  
pp. 147-153 ◽  
Author(s):  
Gabriella Lindergard ◽  
Daryl V. Nydam ◽  
Susan E. Wade ◽  
Stephanie L. Schaaf ◽  
Hussni O. Mohammed
Keyword(s):  
Dna Extraction ◽  
Extraction Methods ◽  
Pcr Detection ◽  
Fecal Samples ◽  
Cryptosporidium Oocysts ◽  
Dna Extraction Methods

scholarly journals Extracting the invisible: obtaining high quality DNA is a challenging task in small arthropods

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6753 ◽  
Author(s):  
Andrea Lienhard ◽  
Sylvia Schäffer
Keyword(s):  
Dna Extraction ◽  
Low Cost ◽  
Extraction Methods ◽  
Chelating Resin ◽  
Precipitation Method ◽  
Ammonium Bromide ◽  
User Friendliness ◽  
Silica Membrane ◽  
High Quality ◽  
Dna Extraction Methods

BackgroundThe application of an appropriate extraction method is a relevant factor for the success of all molecular studies.MethodsSeven different DNA extraction methods suitable for high-throughput DNA sequencing with very small arthropods were compared by applying nine different protocols: three silica gel based spin methods, two cetyltrimethyl ammonium bromide (CTAB) based ones (one with an additional silica membrane), a protein precipitation method and a method based on a chelating resin (applying different protocols). The quantity (concentration) and quality (degradation, contamination, polymerase chain reaction (PCR) and sequencing success) of the extracted DNA as well as the costs, preparation times, user friendliness, and required supplies were compared across these methods. To assess the DNA quantity, two different DNA concentration measurements were applied. Additionally, the effect of varying amounts of starting material (different body sizes), variable lysis temperatures and mixing during DNA extraction was evaluated.ResultsAlthough low DNA concentrations were measured for all methods, the results showed that—with the exception of two methods—the PCR success was 100%. However, other parameters show vast differences. The time taken to perform DNA extraction varied from 20 min to 2.5 h (Chelex vs. CTAB) and the costs from 0.02 to 3.46 € (Chelex vs. QIAamp kit) per sample. High quality genomic DNA was only gained from four methods. Results of DNA quantity measurements further indicated that some devices cannot deal with small amounts of DNA and show variant results.DiscussionIn conclusion, using Chelex (chelating resin) turned out as a rapid, low-cost method which can provide high quality DNA for different kinds of molecular investigations.

scholarly journals Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples

2018 ◽  
Vol 11 (1) ◽  
Author(s):  
Estelle Menu ◽  
Charles Mary ◽  
Isabelle Toga ◽  
Didier Raoult ◽  
Stéphane Ranque ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Extraction Methods ◽  
Enteric Pathogens ◽  
Fecal Samples ◽  
Dna Extraction Methods

scholarly journals Occurrence of gastrointestinal parasites in goat kids

2012 ◽  
Vol 21 (1) ◽  
pp. 65-67 ◽  
Author(s):  
Willian Marinho Dourado Coelho ◽  
Alessandro Francisco Talamini do Amarante ◽  
Katia Denise Saraiva Bresciani
Keyword(s):  
Eimeria Species ◽  
Gastrointestinal Parasites ◽  
Male And Female ◽  
Fecal Samples ◽  
Helminth Eggs ◽  
Eimeria Spp ◽  
One Year

Fecal samples from male and female goat kids, of different breeds and up to one year of age, were analyzed to determine egg and oocyst counts per gram of feces (EPG and OPG, respectively), and fecal culturing was performed to identify nematode genera. Helminth eggs and Eimeria spp. oocysts were found in 93.06% (188/202) and 77.22% (156/202) of the fecal samples, respectively. From fecal cultures, the following genera were identified: Cooperia in 11.88% (24/202), Haemonchus in 51.98% (105/202), Oesophagostomum in 9.4% (19/202), Strongyloides in 5.94 (12/202) and Trichostrongylus in 20.79% (42/202) of the samples. The Eimeria species found were E. alijevi in 25.24% (51/202), E. arloingi in 7.42% (15/202), E. caprina in 2.97% (6/202), E. caprovina in 10.39% (21/202), E. christenseni in 4.45% (9/202), E. joklchijevi in 11.38% (23/202), E. hirci in 9.4% (19/202) and E. ninakohlyakimovae in 28.71% (58/202) samples. Among the gastrointestinal parasites, the genus Haemonchus and two Eimeria species (E. ninakohlyakimovae and E. alijevi) were predominants.

Sign in / Sign up

Export Citation Format

Share Document