scholarly journals Molecular survey of Babesia vogeli and Hepatozoon species in dogs from urban area of Midwestern Brazil

2019 ◽  
Vol 40 (3) ◽  
pp. 1357
Author(s):  
Maerle Oliveira Maia ◽  
André Luís Santos de Freitas ◽  
Jamila Guimarães Santos ◽  
Thábata dos Anjos Pacheco ◽  
Dirceu Guilherme de Souza Ramos ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Urban Areas ◽  
Ixodid Ticks ◽  
Hepatozoon Canis ◽  
Molecular Survey ◽  
Rdna Sequences ◽  
Babesia Vogeli ◽  
18S Rdna Sequences ◽  
Polymerase Chain ◽  
Hepatozoon Species

In Brazil, the most important tickborne pathogens affecting dogs include Babesia vogeli, Ehrlichia canis, Anaplasma platys, Hepatozoon canis, and Mycoplasma haemocanis. Babesia spp. and Hepatozoon spp., transmitted by ixodid ticks, have been reported to naturally infect dogs and are widespread. The authors aimed to investigate the incidence of B. vogeli and Hepatozoon spp. infection using molecular methods to identify factors associated with the infection in dogs from urban areas of Cuiabá municipality, Midwestern Brazil. Polymerase chain reaction (PCR) assay revealed a prevalence of 9.36% (Confidence Interval-CI 95%; 2.72%; 6.79%) and 9.61% (CI 95%; 7.0%; 13.0%) among dogs for B. vogeli and Hepatozoon, respectively. DNA sequences obtained from 10 Hepatozoon PCR positive samples were sequenced and were identical to one another and, moreover, were 100% (541/541 base of pairs-bp) homologous to the corresponding 18S rDNA sequences of H. canis. Twenty-five dogs (6.02%) generated amplicons using PCR protocols for both organisms, indicating co-infection by these two protozoans. To the best of our knowledge, our study was the first molecular survey to consider the entire population of dogs from the study area. Moreover, young dogs (0-12 months of age), as well as animals living in walled houses?without access to the street?were more susceptible to infection with B. vogeli and H. canis, respectively.

scholarly journals SEQUENCE ANALYSIS OF 18s DNA OF Melosira sp., Dunaliella sp., Isochrysis sp. AND Porphyridium sp.

2015 ◽  
Vol 2 (1) ◽  
pp. 592
Author(s):  
Lucia Kusumawati ◽  
Ruben Wahyudi ◽  
Reinhard Pinontoan ◽  
Maria Gorreti Lily Panggabean
Keyword(s):  
Sequence Analysis ◽  
Phylogenetic Tree ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Morphological Characters ◽  
Rdna Sequences ◽  
Pcr Products ◽  
18S Rdna Sequences ◽  
Pcr Method ◽  
High Level

<p>Phytoplankton has high level of biodiversity. In previous years phytoplankton was identified by their morphological characters. However, their morphology might change in different environments. These difficulties can be overcome by comparing their 18S rDNA sequences. This research is aimed to verify the identity of Melosira sp., Dunaliella sp., Isochrysis sp. and Porphyridium sp. Here, PCR method was used to amplify 18s DNA sequences. Three primer pairs were used, i.e. 18S-F and 18S-R; 501F and 1700R; 18S-2F and 18S-2R. PCR products were sequenced. MEGA5 was used to make phylogenetic tree. Genus verification for Isochrysis sp., Dunaliella sp. and Melosira sp. were conducted successfully using Blast and phylogenetic tree. 18s DNA sequence of Porphyridium sp. shows an interesting result and needs further verification.</p><p><br /><strong>Keywords</strong>: Phytoplankton, Melosira sp., Dunaliella sp., Isochrysis sp., Porphyridium sp.</p>

Survey of slug-parasitic nematodes in East and West Flanders, Belgium and description ofAngiostoma gandavensisn. sp. (Nematoda: Angiostomidae) from arionid slugs

2019 ◽  
Vol 94 ◽  
Author(s):  
P.R. Singh ◽  
M. Couvreur ◽  
W. Decraemer ◽  
W. Bert
Keyword(s):  
New Species ◽  
Light Microscopy ◽  
Dna Sequences ◽  
Morphological Characters ◽  
Molecular Data ◽  
Parasitic Nematodes ◽  
Rdna Sequences ◽  
18S Rdna Sequences ◽  
East And West ◽  
Tail Tip

AbstractA survey for slug-associated nematodes in five locations of East and West Flanders in Belgium revealed the presence of one new and six known slug-parasitic nematodes,Agfa flexilis(Dujardin, 1845),Alloionema appendiculatum(Schneider, 1859),Angiostoma dentiferum(Mengert, 1953),Angiostoma limacis(Dujardin, 1845),Angiostoma norvegicum(Rosset al., 2017) andPhasmarhabditis hermaphrodita(Schneider, 1859).Angiostoma norvegicumandP.hermaphroditaare recorded for the first time in Belgium. The six known species are documented by light microscopy (LM) microphotographs and informative DNA sequences.Angiostoma gandavensisn. sp. (Angiostomatidae), discovered from arionid slugs, is described based on light microscopy, scanning electron microscopy (SEM) and molecular data. Based on analyses of D2D3 expansion segment of 28S and 18S rDNA sequences, this new species is found to be related toA.limacis,A.norvegicum,A.margaretae(Rosset al., 2011) andA.milacis(Ivanova and Wilson, 2009). The new species can be distinguished from these others based on morphological characters such as the distinctive mucronate structures at the tail tip of both sexes, presence of lateral ala, reflexed female ovaries and the number and arrangement pattern of male genital papillae.

scholarly journals Monitoring the Bacterial and Fungal Biota of Eleven Tobacco Grades Stored at Three Different Locations

2009 ◽  
Vol 23 (6) ◽  
pp. 368-376
Author(s):  
R Villemur ◽  
M Lacasse ◽  
A Morin
Keyword(s):  
Ribosomal Rna ◽  
Storage Time ◽  
Bacterial Population ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Rdna Sequences ◽  
Fungal Isolates ◽  
Gradient Gel Electrophoresis ◽  
18S Rdna Sequences ◽  
Polymerase Chain

AbstractTobacco as many other plants has its own microbiota. There are very few studies determining the evolution of this microbiota during tobacco storage, which may affect the quality of tobacco. Polymerase chain reaction (PCR) combined with denaturing gradient gel electrophoresis (DGGE) were used to determine changes in the microbiota of tobacco during the aging of eleven different tobacco grades stored at three different locations for twelve months. The microbial fraction of these tobacco grades was extracted, and the bacterial 16S and the fungal 18S ribosomal RNA gene (rDNA) sequences were PCR amplified before being segregated by DGGE. The bacterial complexity of the tobacco grades was represented by DGGE migrating banding profiles that varied between 20 and 30 bands. Some variations in the banding profiles were observed between the tobacco grades, but overall no substantial changes occurred in the bacterial population of the different grades during their storage at different locations. Most of the fungal DGGE profiles were identical and had only one dominating band related to the genus Aspergillus. Bacterial and fungal isolates were also derived from the microbial fractions of the tobacco, and part of their respective 16S and 18S rDNA sequences were determined. Bacterial isolates belonged to Bacillales and gamma Protobacteria. Fungal isolates belonged to the genus Aspergillus. Our results showed that the bacterial and fungal biota of tobacco are relatively stable throughout 12 months storage time.

scholarly journals Genetic diversity of Hepatozoon spp. in rodents from Chile

2021 ◽  
Vol 30 (4) ◽  
Author(s):  
Amir Salvador Alabí ◽  
Gustavo Monti ◽  
Carola Otth ◽  
Paulina Sepulveda-García ◽  
Livia Perles ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
18S Rdna ◽  
Nucleotide Polymorphism ◽  
Conventional Pcr ◽  
Rdna Sequences ◽  
18S Rdna Sequences ◽  
Hepatozoon Species ◽  
First Time ◽  
18S Rdna Gene ◽  
High Diversity

Abstract This study aimed to investigate the genetic diversity of Hepatozoon spp. in rodents from Valdivia, Chile. A total of 74 rodents (synanthropic n=38; wild n=36) were trapped in Valdivia. We performed conventional PCR assays for Apicomplexa organisms targeting two overlapping 18S rDNA gene fragments (600 bp and 900 bp) followed by sequencing of selected amplicons. Hepatozoon spp. occurrence was 82.43% (61/74). Twelve sequences obtained from the 600 bp and ten from the 900 bp 18S rDNA fragments were identified as Hepatozoon sp. Six sequences obtained from 18S rDNA-based overlapping PCR protocols were used for concatenated (1,400 bp) phylogenetic, haplotype and distance analyses. Hepatozoon spp. 18S rDNA concatenated sequences from the present study were detected in Oligoryzomys longicaudatus, Rattus norvegicus, Mus musculus, and Abrothrix longipilis grouped with Hepatozoon species earlier described in rodents and reptiles from Chile and Brazil. Nucleotide polymorphism of the six 18S rDNA sequences (1,400 bp) from this study, and other Chilean sequences from rodents and rodent’s ticks, showed high diversity with a total of nine Chilean haplotypes. Three haplotypes from Valdivia were identified for the first time in this study, suggesting the circulation of novel haplotypes in rodents from southern Chile.

Biodegradation of Phenol by Antarctic Strains of Aspergillus fumigatus

2013 ◽  
Vol 68 (9-10) ◽  
pp. 384-393 ◽  
Author(s):  
Maria Gerginova ◽  
Jordan Manasiev ◽  
Husein Yemendzhiev ◽  
Anna Terziyska ◽  
Nadejda Peneva ◽  
...  
Keyword(s):  
Aspergillus Fumigatus ◽  
Dna Sequences ◽  
Phenol Degradation ◽  
Phenol Hydroxylase ◽  
Sequence Identity ◽  
Rdna Sequences ◽  
Catechol 1,2 Dioxygenase ◽  
18S Rdna Sequences ◽  
Intracellular Activities ◽  
High Degree

Taxonomic identifi cation of three newly isolated Antarctic fungal strains by their 18S rDNA sequences revealed their affi liation with Aspergillus fumigatus. Phenol (0.5 g/l) as the sole carbon source was completely degraded by all strains within less than two weeks. Intracellular activities of three key enzymes involved in the phenol catabolism were determined. Activities of phenol hydroxylase (EC 1.14.13.7), hydroquinone hydroxylase (EC 1.14.13.x), and catechol 1,2-dioxygenase (EC 1.13.11.1) varied signifi cantly between strains. The rates of phenol degradation in the three strains correlated best with the activity of catechol 1,2-dioxygenase. Six pairs of oligonucleotide primers were designed on the basis of the Aspergillus fumigatus Af293 genome sequence (NCBI Acc. No. XM_743491.1) and used to amplify phenol hydroxylase-related gene sequences. DNA sequences of about 1200 bp were amplifi ed from all three strains and found to have a high degree of sequence identity with the corresponding gene of Aspergillus fumigatus Af293

Detection and molecular identification of Hepatozoon canis and Babesia vogeli from domestic dogs in Palestine

Parasitology ◽  
2016 ◽  
Vol 144 (5) ◽  
pp. 613-621 ◽  
Author(s):  
KIFAYA AZMI ◽  
AMER AL-JAWABREH ◽  
ABEDELMAJEED NASEREDDIN ◽  
AHMAD ABDELKADER ◽  
TAHER ZAID ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Rhipicephalus Sanguineus ◽  
18S Rrna Gene ◽  
Blood Parasites ◽  
Hepatozoon Canis ◽  
Rrna Gene ◽  
Babesia Vogeli ◽  
Wide Range ◽  
Pcr Products ◽  
Hepatozoon Felis

SUMMARYDogs serve as hosts for a great number of parasites, which may affect their health and wellbeing. This study aimed to observe tick borne pathogens in dogs from Palestine including Hepatozoon canis and Babesia species. The prevalence of both H. canis and Babesia species infections in apparently healthy dogs, from ten districts of the West Bank was surveyed. DNA was extracted from blood samples obtained from dogs (n = 362) and ticks (n = 213) collected from dogs (n = 77). A primer set that amplifies a partial sequence of the Babesia and Hepatozoon 18S rRNA gene was used for PCR and the DNA sequences of the PCR products of all samples were determined. Twenty-nine (8·0%) of the dogs were found infected including 20 with H. canis (5·5%), seven with Babesia vogeli (1·9%) and two with undefined Babesia spp. (0·6%). Twelve Rhipicephalus sanguineus s.l ticks were pathogen-positive, including ten with H. canis (4·7%), one with B. vogeli (0·5%), and one with Hepatozoon felis (0·5%). The results indicated that a wide range of tick borne pathogens is circulating in the canine population in the surveyed region. This study is the first report on the prevalence of H. canis, B. vogeli and Babesia spp. in dogs in Palestine and its results will assist in the management of diseases associated with these blood parasites.

scholarly journals DMSO Improves the Ski-Slope Effect in Direct PCR

2021 ◽  
Vol 11 (4) ◽  
pp. 1943
Author(s):  
Joo-Young Kim ◽  
Ju Yeon Jung ◽  
Da-Hye Kim ◽  
Seohyun Moon ◽  
Won-Hae Lee ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Pcr Amplification ◽  
Analytical Techniques ◽  
Direct Pcr ◽  
Dna Profile ◽  
Novel Technologies ◽  
Specific Amplification ◽  
Polymerase Chain ◽  
Dna Profiles

Analytical techniques such as DNA profiling are widely used in various fields, including forensic science, and novel technologies such as direct polymerase chain reaction (PCR) amplification are continuously being developed in order to acquire DNA profiles efficiently. However, non-specific amplification may occur depending on the quality of the crime scene evidence and amplification methods employed. In particular, the ski-slope effect observed in direct PCR amplification has led to inaccurate interpretations of the DNA profile results. In this study, we aimed to reduce the ski-slope effect by using dimethyl sulfoxide (DMSO) in direct PCR. We confirmed that DMSO (3.75%, v/v) increased the amplification yield of large-sized DNA sequences more than that of small-sized ones. Using 50 Korean buccal samples, we further demonstrated that DMSO reduced the ski-slope effect in direct PCR. These results suggest that the experimental method developed in this study is suitable for direct PCR and may help to successfully obtain DNA profiles from various types of evidence at crime scenes.

Using a Polymerase Chain Reaction as a Complementary Test to Improve the Detection of Dermatophyte Fungus in Nails

2014 ◽  
Vol 104 (3) ◽  
pp. 233-237 ◽  
Author(s):  
María José Iglesias Sánchez ◽  
Ana María Pérez Pico ◽  
Félix Marcos Tejedor ◽  
María Jesús Iglesias Sánchez ◽  
Raquel Mayordomo Acevedo
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Classical Method ◽  
Clinical Manifestations ◽  
Culture Method ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Traditional Procedure ◽  
Polymerase Chain

Background Dermatomycoses are a group of pathologic abnormalities frequently seen in clinical practice, and their prevalence has increased in recent decades. Diagnostic confirmation of mycotic infection in nails is essential because there are several pathologic conditions with similar clinical manifestations. The classical method for confirming the presence of fungus in nail is microbiological culture and the identification of morphological structures by microscopy. Methods We devised a nested polymerase chain reaction (PCR) that amplifies specific DNA sequences of dermatophyte fungus that is notably faster than the 3 to 4 weeks that the traditional procedure takes. We compared this new technique and the conventional plate culture method in 225 nail samples. The results were subjected to statistical analysis. Results We found concordance in 78.2% of the samples analyzed by the two methods and increased sensitivity when simultaneously using the two methods to analyze clinical samples. Now we can confirm the presence of dermatophyte fungus in most of the positive samples in just 24 hours, and we have to wait for the result of culture only in negative PCR cases. Conclusions Although this PCR cannot, at present, substitute for the traditional culture method in the detection of dermatophyte infection of the nails, it can be used as a complementary technique because its main advantage lies in the significant reduction of time used for diagnosis, in addition to higher sensitivity.

scholarly journals Phylogeny of the Thoracican Barnacles Based on 18S rDNA Sequences

2000 ◽  
Vol 20 (2) ◽  
pp. 393-398 ◽  
Author(s):  
D. James Harris ◽  
Laura S. Maxson ◽  
Lee F. Braithwaite ◽  
Keith A. Crandall
Keyword(s):  
18S Rdna ◽  
Rdna Sequences ◽  
18S Rdna Sequences

Polymerase chain reaction based mapping of rye involving repeated DNA sequences

Genome ◽  
1992 ◽  
Vol 35 (4) ◽  
pp. 621-626 ◽  
Author(s):  
Peter M. Rogowsky ◽  
Ken W. Shepherd ◽  
Peter Langridge
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Chromosome 1 ◽  
Repeated Dna ◽  
Chain Reaction ◽  
Pcr Marker ◽  
Banding Patterns ◽  
Repeated Dna Sequences ◽  
Cereal Rye ◽  
Polymerase Chain

A novel type of polymerase chain reaction (PCR) marker was developed for the mapping of cereal rye (Secale cereale). Primer pairs were synthesized targeting the insertion sites of three individual copies of the R173 family of rye specific repeated DNA sequences. While one primer was derived from a sequence within the respective R173 element, the second primer corresponded to a flanking region. The complex banding patterns obtained in rye allowed not only the mapping of the three R173 elements to certain chromosome regions of 1RS (the short arm of rye chromosome 1) but also the mapping of an additional 3–10 easily identifiable bands per primer pair to other rye chromosomes. Linkage mapping of a polymorphic 1R band derived from three rye cultivars demonstrated the presence of nonallelic, dominant markers in two independent crosses. Because of the high copy number of the R173 family (15 000 copies per diploid rye genome), its dispersion over the entire length of all chromosomes and the high number of markers obtained per primer pair, PCR markers based on the R173 family provide an almost unlimited source for well-spaced markers in rye mapping.Key words: polymerase chain reaction, mapping, repetitive DNA sequences, wheat, rye.

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