scholarly journals Molecular Evidence of Rickettsia felis in Phereoeca sp.

2021 ◽  
Vol 30 (1) ◽  
Author(s):  
Fernanda Sales de Araújo ◽  
Rafael Mazioli Barcelos ◽  
Tiago Antônio de Oliveira Mendes ◽  
Cláudio Mafra
Keyword(s):  
Blood Feeding ◽  
Coi Gene ◽  
Pcr Analysis ◽  
Molecular Evidence ◽  
Rickettsia Felis ◽  
Obligate Intracellular ◽  
Transitional Group ◽  
Polymerase Chain ◽  
Mutualistic Association ◽  
Level Identification

Abstract Rickettsia felis is an obligate intracellular bacterium capable of infecting ticks, fleas, lice, and other arthropods. This bacterium is classified as a member of the Transitional Group (TRG) Rickettsia. It is known the evidence of R. felis mutualistic and obligatory relationship with some eukaryote organisms. However, there aren’t scientific accounts of R. felis and moths of the order Lepidoptera association. The current work reports the first identification of the bacteria R. felis in Phereoeca sp. For that, a polymerase chain reaction (PCR) assay using gltA, ompA, and ompB genes was used. The nucleotide sequences showed 100% of identity with other Rickettsia felis sequences. The genus-level identification of the moth larvae was performed by morphological taxonomic keys and PCR analysis of the cytochrome oxidase I (COI) gene. The nucleotide sequenced showed 94.94% similarity with the species Phereoeca praecox. However, with the low number of sequences deposited in the databases, the species was classified as Phereoeca sp. The results suggest that R. felis may develop in an organism without blood-feeding behavior (Lepidoptera), as it has been demonstrated for booklice (Psocoptera). Further investigation is necessary in order to confirm pathogenic or mutualistic association with moths.

Molecular identification and characterization of Trichinella spiralis from a leopard in India

2019 ◽  
Author(s):  
Anil Kumar Nehra ◽  
Hira Ram ◽  
P. S. Banerjee ◽  
Rajat Garg ◽  
M. Karikalan ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Genetic Markers ◽  
Trichinella Spiralis ◽  
Ribosomal Subunit ◽  
Species Level ◽  
Homology Search ◽  
Coi Gene ◽  
Molecular Evidence ◽  
Level Identification

TPresent study describes species level identification of Trichinella spiralis of leopard origin from India using multiplex PCR and molecular characterization of the parasite based on sequencing of multiple genetic markers viz. 5S ribosomal DNA intergenic spacer region (5S ISR), partial mitochondrial large ribosomal subunit (Mt-lsr) and partial mitochondrial cytochrome c oxidase I (COI) genes. A single amplicons of 173 bp, indicative of T. spiralis was obtained in multiplex PCR. Further, specific PCR amplifications viz. 750 bp (5S ISR), 445 bp (Mt-lsr) and 850 bp (COI) were obtained for selected genetic markers. Homology search analysis of 5S-ISR, Mt-lsr gene and COI gene showed highest 99.6% identity with sequences originating from China (KT894074, T. spiralis), 98.6% similarity with T. spiralis China isolates (GU339127, GU339147) and 99.8% sequence homology with T. spiralis sequences originating from Belarus (MH119334), respectively. In the phylogenetic analysis, sequences of each selected genetic marker clustered together with published T. spiralis isolates only, which further confirmed species level identification of detected larvae as T. spiralis, although very few differences were noted with reference to relative positions. This is the first study from India, which provide molecular evidence on circulation of T. spiralis in wild animals.

scholarly journals Molecular detection of Rickettsia spp. in ticks collected from dogs from the Department of Piura, Peru

2018 ◽  
Vol 39 (2) ◽  
pp. 605
Author(s):  
Luis Fernando Cerro Temoche ◽  
Eloy Da Silva Seabra Junior ◽  
Matheus Dias Cordeiro ◽  
Adivaldo Henrique da Fonseca ◽  
Nathalie Costa da Cunha ◽  
...  
Keyword(s):  
Urban Areas ◽  
Rhipicephalus Sanguineus ◽  
Positive Sample ◽  
Metropolitan Region ◽  
Positive Sequence ◽  
Pcr Analysis ◽  
Rickettsia Felis ◽  
Amblyomma Triste ◽  
Total Dna ◽  
Polymerase Chain

The aim of this study was to determine the frequency of ticks positive for genus Rickettsia bacteria among ticks collected from domestic dogs in the Department of Piura, Peru, using polymerase chain reaction (PCR) analysis. Ticks were collected from dogs in urban areas of the metropolitan region of Piura, Peru. Only three species of ticks were identified; 977 Rhipicephalus sanguineus (180 nymphs, 417 females, and 380 males), Six Amblyomma triste females, and one Amblyomma tigrinum male. After classifying the specimens morphologically by stage, species, and sex, their total DNA was tested by PCR using primers that amplify fragments of the gltA, ompA, ompB, and htrA genes. The resulting positive sample was sequenced, compared to the GenBank database, and analyzed phylogenetically. The Rickettsia spp. infection rate in the tick pools was 0.2% (1/484); the positive specimen was an R. sanguineus tick. GenBank analysis of the positive sequence revealed 100% identify with Rickettsia felis; however, no products of the htrA, ompA and ompB genes were amplified from this sample. To the best of our knowledge, this is the first report of R. felis in R. sanguineus in Peru.

scholarly journals Technical Limitations Associated With Molecular Barcoding of Arthropod Bloodmeals Taken From North American Deer Species

2020 ◽  
Vol 57 (6) ◽  
pp. 2002-2006
Author(s):  
Erin M Borland ◽  
Daniel A Hartman ◽  
Matthew W Hopken ◽  
Antoinette J Piaggio ◽  
Rebekah C Kading
Keyword(s):  
Mitochondrial Gene ◽  
Blood Feeding ◽  
Species Level ◽  
Coi Gene ◽  
Cytochrome C Oxidase I ◽  
Molecular Barcoding ◽  
Deer Species ◽  
Multiple Loci ◽  
D Loop ◽  
Level Identification

Abstract Accurate species-level identification of the source of arthropod bloodmeals is important for deciphering blood feeding patterns of field-collected specimens. Cytochrome c oxidase I (COI) mitochondrial gene sequencing has been used for this purpose; however, species resolution can be difficult to obtain from certain vertebrate genera, including Odocoileus. Sanger sequencing of mitochondrial genes was employed to identify the bloodmeal source of wild-caught mosquitoes trapped in Greeley, Colorado. Initial sequencing of the COI gene of mitochondrial DNA in bloodmeals was inadequate for species-level resolution of bloodmeals from deer in the genus Odocoileus, with current databases returning low fidelity matches to multiple genera. The use of the hypervariable D loop of the control region provided species-level identification of white-tailed deer (Order: Artiodactyla, Family: Cervidae, Odocoileus virginianus); however, taxonomic identification was successful only to genus for mule (O. hemionus hemionus) and black-tailed deer (O. hemionus columbianus). We advocate the use of multiple loci for bloodmeal analysis and the buildout of available databases to include multiple mitochondrial reference genes for reliable host species identification.

Sensitivity of polymerase chain reaction (PCR)-southern hybridization and conventional PCR analysis for Halal authentication of gelatin capsules

LWT ◽  
2015 ◽  
Vol 63 (1) ◽  
pp. 714-719 ◽  
Author(s):  
Sahilah Abd Mutalib ◽  
Nursheila Mustafa Muin ◽  
Aminah Abdullah ◽  
Osman Hassan ◽  
Wan Aida Wan Mustapha ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Southern Hybridization ◽  
Pcr Analysis ◽  
Conventional Pcr ◽  
Chain Reaction ◽  
Gelatin Capsules ◽  
Halal Authentication ◽  
Polymerase Chain

scholarly journals Effect of a Berry Polyphenolic Fraction on Biofilm Formation, Adherence Properties and Gene Expression of Streptococcus mutans and Its Biocompatibility with Oral Epithelial Cells

Antibiotics ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 46
Author(s):  
Mariem Souissi ◽  
Amel Ben Lagha ◽  
Kamel Chaieb ◽  
Daniel Grenier
Keyword(s):  
Epithelial Cells ◽  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Cell Model ◽  
Pcr Analysis ◽  
Adhesion Properties ◽  
Oral Epithelial Cells ◽  
Polymerase Chain ◽  
Oral Epithelial ◽  
Wild Blueberry

The ability of Streptococcus mutans to adhere to oral surfaces and form biofilm is a key step in the tooth decay process. The aim of this study was to investigate a berry (wild blueberry, cranberry, and strawberry) polyphenolic fraction, commercialized as Orophenol®, for its antibacterial, anti-biofilm, and anti-adhesion properties on S. mutans. Moreover, the biocompatibility of the fraction with human oral epithelial cells was assessed. Phenolic acids, flavonoids (flavonols, anthocyanins, flavan-3-ols), and procyanidins made up 10.71%, 19.76%, and 5.29% of the berry polyphenolic fraction, respectively, as determined by chromatography and mass spectrometry. The berry polyphenolic preparation dose-dependently inhibited S. mutans biofilm formation while not reducing bacterial growth. At concentrations ranging from 250 to 1000 µg/mL, the fraction inhibited the adhesion of S. mutans to both saliva-coated hydroxyapatite and saliva-coated nickel–chrome alloy. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis showed that incubating S. mutans with the berry polyphenolic fraction was associated with a reduced expression of luxS gene, which regulates quorum sensing in S. mutans. The berry fraction did not show any significant cytotoxicity in an oral epithelial cell model. In conclusion, Orophenol®, which is a mixture of polyphenols from wild blueberry, cranberry and strawberry, possesses interesting anti-caries properties while being compatible with oral epithelial cells.

DNA barcodes provide evidence for the independent origin of the cultivated silkworms Samia ricini and Samia cynthia

2021 ◽  
pp. 1-8
Author(s):  
J.-C. Huang ◽  
X.-Y. Li ◽  
Y.-P. Li ◽  
R.-S. Zhang ◽  
D.-B. Chen ◽  
...  
Keyword(s):  
Genetic Distance ◽  
Phylogenetic Relationship ◽  
Dna Barcode ◽  
Distinct Species ◽  
Coi Gene ◽  
Molecular Evidence ◽  
Dna Barcodes ◽  
Close Relationship ◽  
Molecular Phylogenetic ◽  
Phylogenetic Framework

Samia ricini (Wm. Jones) and Samia cynthia (Drury) (Lepidoptera: Saturniidae) have been used as traditional sources of food as well as silk-producing insects. However, the phylogenetic relationship between the two silkworms remains to be addressed. In this study, the mitochondrial cytochrome c oxidase subunit 1 (COI) gene sequences corresponding to DNA barcodes from 13 Samia species were analysed, and a DNA barcode-based phylogenetic framework for these Samia species was provided. Phylogenetic analysis showed that multiple individuals of a species could be clustered together. Our analysis revealed a close relationship among Samia yayukae Paukstadt, Peigler and Paukstadt, Samia abrerai Naumann and Peigler, Samia kohlli Naumann and Peigler, Samia naessigi Naumann and Peigler, Samia naumanni Paukstadt, Peigler and Paukstadt, and Samia kalimantanensis Paukstadt and Paukstadt. The mixed clustering relationship and low Kimura-2-parameter (K2P) genetic distance (0.006) between individuals of S. ricini and Samia canningi (Hutton) indicated that the cultivated silkworm S. ricini was derived from the non-cultivated silkworm S. canningi. The remote phylogenetic relationship and high K2P genetic distance (0.039) indicated that S. ricini and S. cynthia are distinct species, thus providing solid molecular evidence that they had entirely independent origins. The relationships between S. kalimantanensis and S. naumanni and between S. cynthia and Samia wangi Naumann and Peigler, as well as the potential cryptic species within S. abrerai were also discussed. This is the first study to assess the DNA barcodes of the genus Samia, which supplements the knowledge of species identification and provides the first molecular phylogenetic framework for Samia species.

Development of Three-Stage Bioaerosol Sampler for Size-Selective Sampling

2022 ◽  
Author(s):  
Jun-Hyung Lim ◽  
Sang Hwan Nam ◽  
Jongwoo Kim ◽  
Nam Hoon Kim ◽  
Gun-Soo Park ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Size Range ◽  
Collection Efficiency ◽  
Commercial Product ◽  
Pcr Analysis ◽  
Cyclone Separator ◽  
Chain Reaction ◽  
Selective Sampling ◽  
Sampling Flow Rate ◽  
Polymerase Chain

Abstract In this study, a three-stage bioaerosol sampler with a sampling flow rate of 170 L/min was designed and fabricated for sampling the bioaerosols released during human breathing and coughing, and its performance was evaluated. The sampler was constructed using a cyclone separator with a cutoff size of 2.5 µm as a preseparator, a multi-nozzle virtual impactor with a cutoff size of 0.34 µm as an aerosol concentrator, and a BioSampler, which is a commercial product, for collecting bioaerosols in a collection fluid. The collection efficiency of the sampler was evaluated through simulations and experiments. Only particles with sizes of 0.1-4 µm were selectively collected in the collection fluid. Bacteriophage bioaerosols were sampled using the developed sampler and ACD-200 Bobcat sampler, which is a commercial product. The amounts of collected bacteriophages were compared using the polymerase chain reaction (PCR) technique. The sampling performance of the developed sampler was similar to that of the ACD-200 Bobcat sampler. Moreover, the developed sampler showed its ability to sample bioaerosols of a specific size-range and collect them directly in a collection fluid for the PCR analysis. Therefore, the developed sampler is expected to be useful for indoor environmental monitoring by effectively sampling the bioaerosols released indoors during human breathing and coughing.

scholarly journals Rickettsia typhi IN RODENTS AND R. felis IN FLEAS IN YUCATÁN AS A POSSIBLE CAUSAL AGENT OF UNDEFINED FEBRILE CASES

2015 ◽  
Vol 57 (2) ◽  
pp. 129-132 ◽  
Author(s):  
Gaspar PENICHE-LARA ◽  
Karla DZUL-ROSADO ◽  
Carlos PÉREZ-OSORIO ◽  
Jorge ZAVALA-CASTRO
Keyword(s):  
Rattus Rattus ◽  
Conventional Polymerase Chain Reaction ◽  
Causal Agent ◽  
Wild Rodents ◽  
Murine Typhus ◽  
Rickettsia Typhi ◽  
Rickettsia Felis ◽  
Vector Borne ◽  
Polymerase Chain ◽  
First Time

Rickettsia typhi is the causal agent of murine typhus; a worldwide zoonotic and vector-borne infectious disease, commonly associated with the presence of domestic and wild rodents. Human cases of murine typhus in the state of Yucatán are frequent. However, there is no evidence of the presence of Rickettsia typhi in mammals or vectors in Yucatán. The presence of Rickettsia in rodents and their ectoparasites was evaluated in a small municipality of Yucatán using the conventional polymerase chain reaction technique and sequencing. The study only identified the presence of Rickettsia typhi in blood samples obtained from Rattus rattus and it reported, for the first time, the presence of R. felis in the flea Polygenis odiosus collected from Ototylomys phyllotis rodent. Additionally, Rickettsia felis was detected in the ectoparasite Ctenocephalides felis fleas parasitizing the wild rodent Peromyscus yucatanicus. This study’s results contributed to a better knowledge of Rickettsia epidemiology in Yucatán.

Characterization of alpha-adrenoceptor gene expression in arterial and venous smooth muscle

1993 ◽  
Vol 265 (5) ◽  
pp. H1501-H1509 ◽  
Author(s):  
P. Ping ◽  
J. E. Faber
Keyword(s):  
Smooth Muscle ◽  
Messenger Rna ◽  
Vena Cava ◽  
Pcr Analysis ◽  
Relative Distribution ◽  
Alpha Adrenoceptor ◽  
Aortic Media ◽  
Unique Restriction ◽  
Polymerase Chain

Six genes coding for three unique alpha 1- (1A, 1B, 1C) and three unique alpha 2- (2A, 2B, 2C) adrenergic receptor (AR) subtypes have been cloned. Ligand binding and contractile studies have demonstrated that both alpha 1- and alpha 2-ARs can exist on vascular smooth muscle (VSM) cells, although less is known about the relative distribution and specific subtypes in different vascular segments. In the present study polymerase chain reaction (PCR) analysis was used to characterize the species of alpha-AR messenger RNA (mRNA) present in freshly isolated rat thoracic aortic media and vena cava and in cultured VSM cells (passage 2) derived from both sources. To prevent possible contamination of VSM mRNA, aortic media was separated from adventitia, and vessels were denuded of endothelial cells. Oligonucleotide primers specific for each of the six adrenergic genes were synthesized and used to probe for the presence of alpha-AR mRNA species after reverse transcription of total cellular RNA to cDNA. PCR-amplified AR transcripts were distinguished by the size of amplified DNA fragments and unique restriction endonuclease cleavage. Expression of alpha 1C- or alpha 2C-mRNA was not detected in vascular tissues or cultured VSM cells, although the alpha 2C-primers detected the expected alpha 2C expression in cerebral cortex. Only alpha 1A-mRNA was detected in aortic adventitia. VSM from aorta expressed alpha 1A-, alpha 1B-, and alpha 2A-mRNA, and this pattern was preserved in cultured aortic VSM. Vena cava also expressed both alpha 1A and alpha 1B; however only alpha 2B-mRNA was detected.(ABSTRACT TRUNCATED AT 250 WORDS)

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