scholarly journals Investigation and molecular identification of Eimeria sp. sampled from captive forest musk deer

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11751
Author(s):  
Ziwei Ren ◽  
Dong Yu ◽  
Wei Zhao ◽  
Yan Luo ◽  
Jianguo Cheng ◽  
...  
Keyword(s):  
Molecular Identification ◽  
18S Rrna ◽  
Pcr Amplification ◽  
Plasmid Vector ◽  
18S Rrna Gene ◽  
Ovis Aries ◽  
Molecular Characteristics ◽  
Rrna Gene ◽  
Musk Deer ◽  
Fecal Samples

Forest musk deer (Moschus berezovskii) is an endangered, protected species in China. Intestinal coccidiosis is a significant problem for captive forest musk deer. However, there are few reports on the prevalence and molecular characteristics of Eimeria sp. in forest musk deer. We sought to investigate the prevalence of Eimeria sp. in forest musk deer in the Sichuan and Shaanxi provinces in China. We also investigated the molecular characteristics of Eimeria sp. by analyzing the 18S rRNA gene. We collected a total of 328 fecal samples from forest musk deer on seven farms throughout the Sichuan and Shaanxi provinces. We extracted this parasite’s DNA and used this as a template for nested PCR amplification. The 18S rRNA gene fragment was associated with the plasmid vector, and these products were introduced into Escherichia coli (DH5α). The cultured bacterial solution was used as a PCR reaction template for identification purposes. We collected 328 fecal samples from forest musk deer in Lixian (n = 54), Maoxian (n = 52), Ma’erkang (n = 49), Dujiangyan (n = 55), Hanyuan (n = 41), Luding (n = 36) and Weinan (n = 41). One hundred ninety-eight (60.37%) fecal samples tested positive for Eimeria sp. . In our analysis of the 18S rRNA gene we found 34 types of Eimeria sp. with a similarity of 90.5–100%. We constructed a phylogenetic tree based on the parasite’s 18S rRNA gene sequence. Our findings indicated that the Eimeria sp. that parasitized the intestinal tract of forest musk deer was closely related to Eimeria alabamensis from Bos taurus and Eimeria ahsata from Ovis aries. To the best of our knowledge, ours was the first investigation and molecular identification of Eimeria sp. sampled from captive forest musk deer in China. Our results provide epidemiological data for the monitoring and prevention of Eimeria sp. in captive forest musk deer.

First report on molecular identification of Eimeria sp. from captive forest musk deer

2020 ◽  
Author(s):  
Ziwei Ren ◽  
Dong Yu ◽  
Wei Zhao ◽  
Yan Luo ◽  
Jianguo Cheng ◽  
...  
Keyword(s):  
18S Rrna ◽  
Intestinal Tract ◽  
Gene Sequence ◽  
18S Rrna Gene ◽  
Ovis Aries ◽  
Rrna Gene ◽  
Gene Fragment ◽  
Musk Deer ◽  
Rrna Gene Sequence ◽  
Faecal Samples

Abstract Background: Forest musk deer (Moschus berezovskii) is a national first-level protected and endangered wild animals in China. The intestinal coccidiosis of captive forest musk deer is one of the most important diseases. However, few studies have been conducted to quantify Eimeria sp. infection and to identify its molecules. Thus, the objective of this study was to investigate the Eimeria sp. infection in the intestinal tract of forest musk deer in Sichuan and Shaanxi, China, and to identify the 18S rRNA gene fragment of Eimeria sp. , which provides scientific basic experimental data for the molecular epidemiological investigation and population genetic analysis of Eimeria sp. of captive forest musk deer in 7 regions.Methods: 328 faecal samples of forest musk deer were collected from 7 farms. The DNA of Eimeria sp. in the positive samples was extracted and used as template for nested PCR amplification. The 18S rRNA gene fragment was connected with the plasmid vector, and the products were introduced into Escherichia coli (DH5α). The culture bacterial solution was used as a PCR reaction template for identification.Results: In total, we collected 328 faecal samples from forest musk deer in Lixian (n=54), Maoxian (n=52), Ma’erkang (n=49), Dujiangyan (n=55), Hanyuan (n=41), Luding (n=36) and Weinan (n=41). 198 (60.37%) faecal samples were positive for Eimeria sp. We analyzed the 18S rRNA gene sequence of Eimeria sp, and determined 34 types with similarity of 90.5% ~ 100%. A phylogenetic tree constructed based on the 18S rRNA gene sequence of Eimeria sp., it was confirmed that Eimeria sp. parasitized in the intestinal tract of forest musk deer was closely related to Eimeria alabamensis from Bos taurus, Eimeria faurei from Ovis aries and Eimeria ahsata from Ovis aries.Conclusions: To our knowledge, this was the first molecular identification of Eimeria sp. in the intestinal tract of forest musk deer. The study suggested that Eimeria sp. parasitizes in the intestinal tract of forest musk deer, which is their carrier.

scholarly journals High-resolution molecular identification of smalltooth sawfish prey

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Taylor L. Hancock ◽  
Gregg R. Poulakis ◽  
Rachel M. Scharer ◽  
S. Gregory Tolley ◽  
Hidetoshi Urakawa
Keyword(s):  
18S Rrna ◽  
Stomach Content Analysis ◽  
18S Rrna Gene ◽  
Taxonomic Resolution ◽  
Molecular Technique ◽  
Rrna Gene ◽  
Fecal Samples ◽  
Predator Prey ◽  
Critically Endangered Species ◽  
Wide Range

AbstractThe foundation of food web analysis is a solid understanding of predator-prey associations. Traditional dietary studies of fishes have been by stomach content analysis. However, these methods are not applicable to Critically Endangered species such as the smalltooth sawfish (Pristis pectinata). Previous research using the combination of stable isotope signatures from fin clips and 18S rRNA gene sequencing of fecal samples identified the smalltooth sawfish as piscivorous at low taxonomic resolution. Here, we present a high taxonomic resolution molecular technique for identification of prey using opportunistically acquired fecal samples. To assess potential biases, primer sets of two mitochondrial genes, 12S and 16S rRNA, were used alongside 18S rRNA, which targets a wider spectrum of taxa. In total, 19 fish taxa from 7 orders and 11 families native to the Gulf of Mexico were successfully identified. The sawfish prey comprised diverse taxa, indicating that this species is a generalist piscivore. These findings and the molecular approach used will aid recovery planning for the smalltooth sawfish and have the potential to reveal previously unknown predator-prey associations from a wide range of taxa, especially rare and hard to sample species.

A new set of primers directed to 18S rRNA gene for molecular identification of Cryptosporidium spp. and their performance in the detection and differentiation of oocysts shed by synanthropic rodents

2013 ◽  
Vol 135 (3) ◽  
pp. 551-557 ◽  
Author(s):  
Sheila O.S. Silva ◽  
Leonardo J. Richtzenhain ◽  
Iracema N. Barros ◽  
Alessandra M.M. C. Gomes ◽  
Aristeu V. Silva ◽  
...  
Keyword(s):  
Molecular Identification ◽  
18S Rrna ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Cryptosporidium Spp ◽  
Synanthropic Rodents

scholarly journals MORPHOLOGICAL AND MOLECULAR VARIATION OF FUSARIUM OXYSPORUM F.SP LYCOPERSICI ISOLATES CAUSING FUSARIUM WILT IN TOMATO

2020 ◽  
Vol 21 (supplement 1) ◽  
Author(s):  
K. Vignesh ◽  
K. Rajamohan ◽  
R. Anandan ◽  
R. Udhayakumar
Keyword(s):  
Fusarium Oxysporum ◽  
Fusarium Wilt ◽  
18S Rrna ◽  
Sequence Similarity ◽  
Fusarium Species ◽  
Critical Role ◽  
Pcr Amplification ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Molecular Variation

Tomato (Solanum lycopersicum L.) is one of the most important, commercial and widely grown vegetable crop in the world. Tomato plays a critical role in nutritional food requirements, income and employment opportunities for the people. However, its production is threatened by the Fusarium wilt caused by Fusarium oxysporum f.sp. lycopersici and productionlossesbetween30%to40%. In the present investigation an attempt has been made to study the morphological and molecular variation of Fusarium oxysporum f.sp lycopersici isolates. Usual identification of Fusarium species based on their micro and macroscopic features and morphological characters alone may lead to incorrect designation. In order to identify the correct species, we amplified the 18S rRNA gene region by PCR, sequenced and analyzed for sequence similarity among the NCBI data through BLAST. Further, PCR amplification of ITS regions was performed using ITS primers. The amplified product of 18S rRNA gene was sequenced and deposited to Gen Bank with the accession numbers.

scholarly journals Molecular survey of Cryptosporidium spp. in calves from the state of Mato Grosso, Brazil

2020 ◽  
Vol 41 (5supl1) ◽  
pp. 2437-2444
Author(s):  
Thábata dos Anjos Pacheco ◽  
Felippe Danyel Cardoso Martins ◽  
Sayanne Luns Hatum de Almeida ◽  
Thiago Borges Fernandes Semedo ◽  
Michelle Igarashi Watanabe ◽  
...  
Keyword(s):  
18S Rrna ◽  
18S Rrna Gene ◽  
The State ◽  
Severe Illness ◽  
Rrna Gene ◽  
Mato Grosso ◽  
Fecal Samples ◽  
Cryptosporidium Spp ◽  
Gp60 Gene ◽  
Wide Range

Cryptosporidium spp. is a protozoan that infects a wide range of vertebrate hosts; it has been reported to be the cause of severe illness or death in livestock worldwide, which leads to decreased performance and production losses, especially in young animals. This study investigated the presence of Cryptosporidium in calves from beef farms in the state of Mato Grosso, midwestern Brazil. For this purpose, fecal samples from 237 animals aged ? 45 days, raised in 20 rural properties were subjected to DNA extraction and nested polymerase chain reaction (nPCR) targeting 18S ribosomal RNA (18S rRNA) gene followed by sequencing. Additionally, positive samples, previously identified as Cryptosporidium parvum by sequencing and phylogenetic analyses based on 18S rRNA gene, were subsequently analyzed focusing the amplification and sequencing using nPCR of a fragment of the 60 kDa glycoprotein (gp60) gene. Of the 237 fecal samples analyzed by PCR (18S rRNA), 50 (21.1%) fecal samples were positive for Cryptosporidium spp., while 14 (70%) of the 20 properties had at least one positive animal. The following Cryptosporidium species were detected: C. bovis, C. parvum, and C. ryanae. Thereafter, two potentially zoonotic subtypes (IIaA15G2R1 and IIaA16G3R1) of C. parvum were identified based on gp60 gene sequences. This study resulted in the detection of subtype IIaA16G3R1 for the first time in South America and showed a wide distribution of the protozoan in beef farms in the studied area of the State.

Host specificity of Trypanosoma (Herpetosoma) species: evidence that bank voles (Clethrionomys glareolus) carry only one T. (H.) evotomys 18S rRNA genotype but wood mice (Apodemus sylvaticus) carry at least two polyphyletic parasites

Parasitology ◽  
2002 ◽  
Vol 124 (2) ◽  
pp. 185-190 ◽  
Author(s):  
H. A. NOYES ◽  
P. AMBROSE ◽  
F. BARKER ◽  
M. BEGON ◽  
M. BENNET ◽  
...  
Keyword(s):  
Host Specificity ◽  
18S Rrna ◽  
Pcr Amplification ◽  
Wood Mouse ◽  
18S Rrna Gene ◽  
Apodemus Sylvaticus ◽  
Clethrionomys Glareolus ◽  
Rrna Gene ◽  
Wood Mice ◽  
Bank Voles

The strongest evidence for host specificity of mammalian trypanosomes comes from parasites of the subgenus Trypanosoma (Herpetosoma). Laboratory studies have shown that T. (Herpetosoma) species will not infect an alternative host. However, this has not been demonstrated in wild populations. We screened 560 bank voles (Clethrionomys glareolus) and 148 wood mice (Apodemus sylvaticus) for trypanosomes by PCR amplification of the 18S rRNA gene. In total, 109 (19%) bank voles and 12 (8%) wood mice were infected. A HaeIII restriction site was discovered that could be used to discriminate between T. (H.) evotomys of the bank vole and T. (H.) grosi of the wood mouse. All the parasites in the bank voles were identified as T. (Herpetosoma) evotomys by RFLP-PCR. Out of the 12 wood mouse infections 10 were due to T. grosi. Two of the wood mice were infected with parasites with a novel genotype that was most similar to those of T. evotomys and T. microti of voles. Fifty-six fleas collected from the rodents were also screened for trypanosomes; 9 were infected with T. evotomys and 1 with T. grosi. One of the fleas infected with T. evotomys was collected from a wood mouse.

scholarly journals Development of a Dinoflagellate-Oriented PCR Primer Set Leads to Detection of Picoplanktonic Dinoflagellates from Long Island Sound

2006 ◽  
Vol 72 (8) ◽  
pp. 5626-5630 ◽  
Author(s):  
Senjie Lin ◽  
Huan Zhang ◽  
Yubo Hou ◽  
Lilibeth Miranda ◽  
Debashish Bhattacharya
Keyword(s):  
18S Rrna ◽  
Long Island ◽  
Pcr Amplification ◽  
Long Island Sound ◽  
Systematic Investigation ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Island Sound ◽  
Pcr Primer

ABSTRACT We developed dinoflagellate-specific 18S rRNA gene primers. PCR amplification using these oligonucleotides for a picoplanktonic DNA sample from Long Island Sound yielded 24 clones, and all but one of these clones were dinoflagellates primarily belonging to undescribed and Amoebophrya-like lineages. These results highlight the need for a systematic investigation of picodinoflagellate diversity in both coastal and oceanic ecosystems.

scholarly journals A Versatile qPCR for Diagnosis of Leporid Gammaherpesvirus 5 Using Evagreen® or Taqman® Technologies

Viruses ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 715
Author(s):  
Fábio A. Abade dos Santos ◽  
Carina L. Carvalho ◽  
Maria C. Peleteiro ◽  
Francisco Parra ◽  
Margarida D. Duarte
Keyword(s):  
Sensitivity And Specificity ◽  
Internal Control ◽  
18S Rrna ◽  
Pcr Amplification ◽  
18S Rrna Gene ◽  
Myxoma Virus ◽  
Rrna Gene ◽  
Molecular Method ◽  
The Real ◽  
Iberian Hare

In late 2019, the first herpesvirus in the genus Lepus, named leporid gammaherpesvirus 5 (LeHV-5) was described. At the time, herpetic typical lesions were observed in hares infected by the myxoma virus, which is known to induce immunosuppression. Though the real impact of LeHV-5 is still poorly understood, since it affects reproduction, it poses an additional threat to the already fragile populations of Iberian hare, demanding prevalence investigations. In this article, we describe the first quantitative molecular method for LeHV-5 detection, using either Taqman or the EvaGreen systems. This method has excellent sensitivity and specificity, it is able to detect 2.1 copies of LeHV-5 DNA and was validated with an internal control targeting the 18S rRNA gene, allowing monitoring extraction and PCR amplification efficiencies.

scholarly journals Molecular characterization of Cryptosporidium xiaoi in goat kids in Bangladesh by nested PCR amplification of 18S rRNA gene

2015 ◽  
Vol 5 (3) ◽  
pp. 202-207 ◽  
Author(s):  
AMAM Zonaed Siddiki ◽  
Sohana Akter Mina ◽  
Zinat Farzana ◽  
Bibi Ayesa ◽  
Rasel Das ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Nested Pcr ◽  
18S Rrna ◽  
Pcr Amplification ◽  
18S Rrna Gene ◽  
Rrna Gene

scholarly journals Phylogenetic Analysis of the Hypervariable Region of the 18S rRNA Gene of Cryptosporidium Oocysts in Feces of Canada Geese (Branta canadensis): Evidence for Five Novel Genotypes

2004 ◽  
Vol 70 (1) ◽  
pp. 452-458 ◽  
Author(s):  
Kristen L. Jellison ◽  
Daniel L. Distel ◽  
Harold F. Hemond ◽  
David B. Schauer
Keyword(s):  
Genetic Diversity ◽  
Phylogenetic Analysis ◽  
18S Rrna ◽  
Hypervariable Region ◽  
Pcr Amplification ◽  
The United States ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Canada Geese ◽  
Pcr Products

ABSTRACT To assess genetic diversity in Cryptosporidium oocysts from Canada geese, 161 fecal samples from Canada geese in the United States were analyzed. Eleven (6.8%) were positive for Cryptosporidium spp. following nested PCR amplification of the hypervariable region of the 18S rRNA gene. Nine PCR products from geese were cloned and sequenced, and all nine diverged from previously reported Cryptosporidium 18S rRNA gene sequences. Five sequences were very similar or identical to each other but genetically distinct from that of Cryptosporidium baileyi; two were most closely related to, but genetically distinct from, the first five; and two were distinct from any other sequence analyzed. One additional sequence in the hypervariable region of the 18S rRNA gene isolated from a cormorant was identical to that of C. baileyi. Phylogenetic analysis provided evidence for new genotypes of Cryptosporidium species in Canada geese. Results of this study suggest that the taxonomy of Cryptosporidium species in geese is complex and that a more complete understanding of genetic diversity among these parasites will facilitate our understanding of oocyst sources and species in the environment.

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