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

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.

2020 ◽  
Vol 21 (supplement 1) ◽  
Author(s):  
K. Vignesh ◽  
K. Rajamohan ◽  
R. Anandan ◽  
R. Udhayakumar

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.


Parasitology ◽  
2002 ◽  
Vol 124 (2) ◽  
pp. 185-190 ◽  
Author(s):  
H. A. NOYES ◽  
P. AMBROSE ◽  
F. BARKER ◽  
M. BEGON ◽  
M. BENNET ◽  
...  

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.


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

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.


1998 ◽  
Vol 30 (4-5) ◽  
pp. 455-462 ◽  
Author(s):  
Katarina Winka ◽  
Carina Ahlberg ◽  
Ove E. Eriksson

AbstractOstropales, in a wide sense, currently encompasses Ostropales s. str., with a few lichenized genera, and Graphidales, most of which are lichenized. Conotrema is the only lichenized genus undoubtedly placed in Ostropales s. str. We have sequenced the 18S rRNA gene from some key taxa to answer the questions as to whether one ortwo orders should be recognized and whether Conotrema should be kept in Ostropales if Graphidales is recognized. We have new sequences from Conotrema populorum (Stictidaceae, Ostropales), Cyanodermella viridula (Stictidaceae Ostropales), Graphis scripta (Graphidaceae Graphidales) and Diploschistes ocellatus var. almeriensis (Thelotremataceae Graphidales). In an analysis with 41 taxa, thelichenized D. ocellatus var. almeriensis and G. scripta are the sister group of the lichenized C. populorum and the non-lichenized C. viridula and Stictis radiata. Together they form a monophyletic cluster supported by a bootstrap value of 82%. Therefore, it is possible to use the ordinal name Ostropales in a wide sense to include the Graphidales. If Ostropales are conceived in a narrow sense, Conotrema should be kept in the Stictidaceae. Eight new PCR primer sequences are reported.


2015 ◽  
Vol 5 (3) ◽  
pp. 202-207 ◽  
Author(s):  
AMAM Zonaed Siddiki ◽  
Sohana Akter Mina ◽  
Zinat Farzana ◽  
Bibi Ayesa ◽  
Rasel Das ◽  
...  

2004 ◽  
Vol 70 (1) ◽  
pp. 452-458 ◽  
Author(s):  
Kristen L. Jellison ◽  
Daniel L. Distel ◽  
Harold F. Hemond ◽  
David B. Schauer

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.


PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11751
Author(s):  
Ziwei Ren ◽  
Dong Yu ◽  
Wei Zhao ◽  
Yan Luo ◽  
Jianguo Cheng ◽  
...  

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.


2021 ◽  
Vol 91 (3) ◽  
pp. 269-276
Author(s):  
Devina Sharma ◽  
◽  
Nirbhay K. Singh ◽  
Harkirat Singh ◽  
Shitanshu S. Rath ◽  
...  

The genetic diversity was studied of Cryptosporidium scrofarum (syn Cryptosporidium pig genotype II) of domestic pigs (Sus scrofa domesticus) from Punjab, India. Nested PCR amplification targeting the 18S rRNA and actin gene loci from Cryptosporidium positive samples was carried out, and the amplicons were sequenced. Phylogenetic comparison of a partial 18S rRNA gene revealed that they were genetically most similar to C. scrofarum isolated from other parts of the world. However, comparison of sequences representing a fragment of the genomic actin locus identified a new genotype conserved within the isolates sampled from India but distinct from other published sequences, suggesting the presence of a different Indian genotype.


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