scholarly journals The pCS20 PCR assay for Ehrlichia ruminantium does not cross-react with the novel deer ehrlichial agent found in white-tailed deer in the United States of America

2004 ◽  
Vol 71 (2) ◽  
Author(s):  
S.M. Mahan ◽  
B.H. Simbi ◽  
M.J. Burridge
Keyword(s):  
United States ◽  
High Specificity ◽  
Pcr Primers ◽  
The United States ◽  
Rrna Gene ◽  
The Novel ◽  
Pcr Assay ◽  
Ehrlichia Ruminantium ◽  
Cowdria Ruminantium ◽  
Specificity And Sensitivity

White-tailed deer are susceptible to heartwater (Ehrlichia [Cowdria] ruminantium infection) and are likely to suffer high mortality if the disease spreads to the United States. It is vital, therefore, to validate a highly specific and sensitive detection method for E. ruminantium infection that can be reliably used in testing white-tailed deer, which are reservoirs of antigenically or genetically related agents such as Ehrlichia chaffeensis, Anaplasma (Ehrlichia) phagocytophilum (HGE agent) and Ehrlichia ewingii. Recently, a novel but as yet unnamed ehrlichial species, the white-tailed deer ehrlichia (WTDE), has been discovered in deer populations in the United States. Although the significance of WTDE as a pathogen is unknown at present, it can be distinguished from other Ehrlichia spp. based on 16S rRNA gene sequence analysis. In this study it was differentiated from E. ruminantium by the use of the pCS20 PCR assay which has high specificity and sensitivity for the detection of E. ruminantium. This assay did not amplify DNA from the WTDE DNA samples isolated from deer resident in Florida, Georgia and Missouri, but amplified the specific 279 bp fragment from E. ruminantium DNA. The specificity of the pCS20 PCR assay for E. ruminantium was confirmed by Southern hybridization. Similarly, the 16S PCR primers (nested) that amplify a specific 405-412 bp fragment from the WTDE DNA samples, did not amplify any product from E. ruminantium DNA. This result demonstrates that it would be possible to differentiate between E. ruminantium and the novel WTDE agent found in white tailed deer by applying the two respective PCR assays followed by Southern hybridizations. Since the pCS20 PCR assay also does not amplify any DNA products from E. chaffeensis or Ehrlichia canis DNA, it is therefore the method of choice for the detection of E. ruminantium in these deer and other animal hosts.

Breaking Free

2017 ◽  
Author(s):  
Deirdre David
Keyword(s):  
United States ◽  
The United States ◽  
The Novel ◽  
Male Homosexuality ◽  
Domestic Life ◽  
The Family

In the mid- to late 1950s, Pamela emerged as a critically acclaimed novelist, particularly after the family returned to London. In perhaps her best-known novel, The Unspeakable Skipton, she explores the life of a paranoid writer who sponges on English visitors to Bruges. The novel was hailed for its wit and sensitive depiction of the life of a writer. She also published a fine study of a London vicar martyred in marriage to a vain and selfish wife: The Humbler Creation is remarkable for its incisive and empathetic depiction of male despair. The Last Resort sealed her distinction as a brilliant novelist of domestic life in its frank depiction of male homosexuality. While continuing to publish fiction, Pamela maintained her reputation as a deft reviewer. In 1954, she and Charles travelled to the United States—the first of many trips that were to follow.

scholarly journals Development of a Co-Dominant Cleaved Amplified Polymorphic Sequences Assay for the Rapid Detection and Differentiation of Two Pathogenic Clarireedia spp. Associated with Dollar Spot in Turfgrass

Agronomy ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1489
Author(s):  
Tammy Stackhouse ◽  
Sumyya Waliullah ◽  
Alfredo D. Martinez-Espinoza ◽  
Bochra Bahri ◽  
Emran Ali
Keyword(s):  
Pcr Primers ◽  
The United States ◽  
Fungal Species ◽  
Dollar Spot ◽  
Direct Pcr ◽  
Specific Pcr ◽  
Specificity And Sensitivity ◽  
Pure Cultures ◽  
Speed Up ◽  
Cleaved Amplified Polymorphic Sequences

Dollar spot is one of the most destructive diseases in turfgrass. The causal agents belong to the genus Clarireedia, which are known for causing necrotic, sunken spots in turfgrass that coalesce into large damaged areas. In low tolerance settings like turfgrass, it is of vital importance to rapidly detect and identify the pathogens. There are a few methods available to identify the genus Clarireedia, but none of those are rapid enough and characterize down to the species level. This study produced a co-dominant cleaved amplified polymorphic sequences (CAPS) test that differentiates between C. jacksonii and C. monteithiana, the two species that cause dollar spot disease within the United States. The calmodulin gene (CaM) was targeted to generate Clarireedia spp. specific PCR primers. The CAPS assay was optimized and tested for specificity and sensitivity using DNA extracted from pure cultures of two Clarireedia spp. and other closely related fungal species. The results showed that the newly developed primer set could amplify both species and was highly sensitive as it detected DNA concentrations as low as 0.005 ng/µL. The assay was further validated using direct PCR to speed up the diagnosis process. This drastically reduces the time needed to identify the dollar spot pathogens. The resulting assay could be used throughout turfgrass settings for a rapid and precise identification method in the US.

scholarly journals Dermacentor variabilis is the Predominant Dermacentor spp. (Acari: Ixodidae) Feeding on Dogs and Cats Throughout the United States

2021 ◽  
Author(s):  
Kathryn T Duncan ◽  
Meriam N Saleh ◽  
Kellee D Sundstrom ◽  
Susan E Little
Keyword(s):  
United States ◽  
Spotted Fever ◽  
Rocky Mountain ◽  
The United States ◽  
Dermacentor Variabilis ◽  
Western United States ◽  
Its2 Region ◽  
Rrna Gene ◽  
Rocky Mountain Spotted Fever ◽  
Dermacentor Albipictus

Abstract Throughout North America, Dermacentor spp. ticks are often found feeding on animals and humans, and are known to transmit pathogens, including the Rocky Mountain spotted fever agent. To better define the identity and distribution of Dermacentor spp. removed from dogs and cats in the United States, ticks submitted from 1,457 dogs (n = 2,924 ticks) and 137 cats (n = 209 ticks) from veterinary practices in 44/50 states from February 2018-January 2020 were identified morphologically (n = 3,133); the identity of ticks from regions where Dermacentor andersoni (Stiles) have been reported, and a subset of ticks from other regions, were confirmed molecularly through amplification and sequencing of the ITS2 region and a 16S rRNA gene fragment. Of the ticks submitted, 99.3% (3,112/3,133) were Dermacentor variabilis (Say), 0.4% (12/3,133) were D. andersoni, and 0.3% (9/3,133) were Dermacentor albipictus (Packard). While translocation of pets prior to tick removal cannot be discounted, the majority (106/122; 87%) of Dermacentor spp. ticks removed from dogs and cats in six Rocky Mountain states (Montana, Idaho, Wyoming, Nevada, Utah, and Colorado) were D. variabilis, suggesting this species may be more widespread in the western United States than is currently recognized, or that D. andersoni, if still common in the region, preferentially feeds on hosts other than dogs and cats. Together, these data support the interpretation that D. variabilis is the predominant Dermacentor species found on pets throughout the United States, a finding that may reflect recent shifts in tick distribution.

scholarly journals Real-Time PCR Assay for Detection and Enumeration of Dekkera bruxellensis in Wine

2003 ◽  
Vol 69 (12) ◽  
pp. 7430-7434 ◽  
Author(s):  
Trevor G. Phister ◽  
David A. Mills
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Primers ◽  
Rrna Gene ◽  
Dekkera Bruxellensis ◽  
Pcr Assay ◽  
Specific Pcr ◽  
Spoilage Yeast ◽  
Pcr Method ◽  
26S Rrna

ABSTRACT Traditional methods to detect the spoilage yeast Dekkera bruxellensis from wine involve lengthy enrichments. To overcome this difficulty, we developed a quantitative real-time PCR method to directly detect and enumerate D. bruxellensis in wine. Specific PCR primers to D. bruxellensis were designed to the 26S rRNA gene, and nontarget yeast and bacteria common to the winery environment were not amplified. The assay was linear over a range of cell concentrations (6 log units) and could detect as little as 1 cell per ml in wine. The addition of large amounts of nontarget yeasts did not impact the efficiency of the assay. This method will be helpful to identify possible routes of D. bruxellensis infection in winery environments. Moreover, the time involved in performing the assay (3 h) should enable winemakers to more quickly make wine processing decisions in order to reduce the threat of spoilage by D. bruxellensis.

scholarly journals A Survey of Genetic Variation in Streptomyces Isolates Causing Potato Common Scab in the United States

2006 ◽  
Vol 96 (12) ◽  
pp. 1363-1371 ◽  
Author(s):  
Leslie A. Wanner
Keyword(s):  
United States ◽  
Genetic Variation ◽  
Common Scab ◽  
Disease Incidence ◽  
The United States ◽  
Rrna Gene ◽  
Population Genetic Study ◽  
Variable Regions ◽  
Genus Streptomyces ◽  
Characteristic Features

Common scab is a serious disease of potatoes and other root and tuber crops, affecting crop quality and market value. The disease is caused by gram positive soil bacteria in the genus Streptomyces. Disease incidence and severity vary in different locations and years; this is due in part to variation in the environment (weather) and genetic variation in potato cultivars. Little information is available on the contribution of genetic variation by the pathogen. To examine genetic diversity in different locations within the United States, streptomycetes were isolated from lesions on field-grown potatoes from six states. Isolates were classified into species based on sequence of variable regions in the 16s rRNA gene. The presence of genes associated with the recently described S. turgidiscabies pathogenicity island (PAI) was also determined. About half of the isolates belonged to S. scabies or S. europaeiscabiei based on 16s rDNA sequence, and had characteristic features of the PAI. They were found in all six states, and were pathogenic on potato and radish. The remaining isolates included pathogens and nonpathogens. They were varied in appearance, and represent several species, including one pathogenic species not previously reported. Some pathogenic isolates lacked one or more genes characteristic of the PAI, although all had genes for biosynthesis of the pathogenicity determinant thaxtomin. In this relatively small survey, regional differences in scab-causing streptomycetes were seen. This report furnishes tools and baseline data for population genetic study of scab-causing streptomycetes in the United States.

Afghanistan Meets the Amazon: Reading The Kite Runner in America

PMLA ◽  
2009 ◽  
Vol 124 (1) ◽  
pp. 25-43 ◽  
Author(s):  
Timothy Aubry
Keyword(s):  
United States ◽  
Military Intervention ◽  
The United States ◽  
The Novel ◽  
United States Military ◽  
War On Terrorism ◽  
Global War On Terrorism ◽  
Customer Reviews ◽  
Global War ◽  
Fictional Representations

This essay considers the American reception of Khaled Hosseini's The Kite Runner in the context of the Bush administration's global war on terrorism by examining the customer reviews of the novel posted on Amazon. As many of the responses indicate, identification serves as a paradoxical means of negotiating with fictional representations of foreignness. The intense and painful empathy inspired by The Kite Runner serves a valorizing function for American readers, strengthening their sense of their own humanity—an effect that resists strict political categorization. Hosseini's ambivalent conception of what it means to be human, I argue, supports a diversity of competing attitudes toward the United States' military intervention in the Middle East and central Asia, while simultaneously catering to fantasies of escape from ideological and cultural divisions altogether.

An Irishman in America: Irishness and Belonging in Roddy Doyle’s Oh, Play That Thing

2021 ◽  
Vol 5 (1) ◽  
Author(s):  
Genoveffa Giambona ◽  
Keyword(s):  
United States ◽  
Identity Construction ◽  
Historical Fiction ◽  
Minority Groups ◽  
The United States ◽  
The Novel

The purpose of this article is to analyse Roddy Doyle’s representations of Irishness and Ireland in Oh, Play That Thing (2004). The novel is the second instalment in Doyle’s The Last Roundup Trilogy, a historical fiction describing the making of the Irish nation through the adventures and misadventures of Henry Smart, its protagonist. In the novel, constructions of Irishness are projected onto the outside world through Henry’s picaresque travels in the United States. The article examines how Irishness is constructed in the book and how it becomes intertwined with identity construction in other minority groups.

scholarly journals Other People’s Cabins: German Inversions of Onkel Tom’s Hutte

Lateral ◽  
2015 ◽  
Vol 4 (1) ◽  
Author(s):  
Kristin Moriah
Keyword(s):  
United States ◽  
20Th Century ◽  
The United States ◽  
Early 20Th Century ◽  
The Novel ◽  
Housing Projects ◽  
Uncle Tom's Cabin ◽  
The Us ◽  
Archival Work

Kristin Moriah’s essay is rooted in extensive archival work in the US and Germany, examining the transatlantic circulation of Uncle Tom’s Cabin through markets of performance and literature in and between Germany and the United States. The essay follows the performative tropes of Uncle Tom’s Cabin from its originary political resonances to the present-day restaurants, train-stops, and housing projects named for the novel. Moriah reveals how the figurations of blackness arising from these texts are foundational to the construction of Germanness and American-German relations in the early 20th century and beyond.

scholarly journals First Report of Root-Knot Nematode Meloidogyne enterolobii on Poinsettia ‘Luv U Pink’ in Taiwan

Plant Disease ◽  
2021 ◽  
Author(s):  
Che-Chang Liang ◽  
P. Janet Chen
Keyword(s):  
United States ◽  
18S Rrna ◽  
The United States ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
First Report ◽  
Meloidogyne Enterolobii ◽  
Mock Control ◽  
Haplotype 1 ◽  
Coii Region

Poinsettia (Euphorbia pulcherrima Willd. ex Klotzsch.), originated in southern Mexico and northern Guatemala, is the most valuable potted flowering plant in the spurge family (Euphorbiaceae). The European Union and the United States are two biggest poinsettia markets (Taylor et al. 2011), with a wholesale value of $153 million in the United States in 2019. Root knot galls of poinsettia ‘Luv U Pink’ were collected from a production greenhouse located in Nantou County, Taiwan in March 2021. No aboveground symptoms were observed. A nematode population was established from a single female and used for identification and the Koch’s postulate. The perineal patterns of randomly picked 5 females are round or ovoid with moderate to high dorsal arches, but no distinct lateral lines, ventral striae are fine and smooth. The Morphometric characters of second-stage juvenile include: a vermiform body shape, tail narrow and tapering with rounded tail tips, and a distinct hyaline tail end. Measurements of 20 J2 are as follows: body length, 430 (398 - 473) μm; body width, 15.4 (13.4 - 17.8) μm; stylet length,13.4 (13.0 - 14.0) μm; dorsal esophageal gland orifice to basal knob, 3.4 (2.8 - 3.9) μm; tail length, 52.9 (47.6 - 62.2) μm. All morphometric data were consistent with the original description of Meloidogyne enterolobii (Yang and Eisenback 1983). Nematode DNA was extracted using GeneMark Tissue & Cell Genomic DNA Purification Kit (GeneMark, Taiwan) from approximately 1500 J2 and used for amplification of 18S rRNA gene, a D2-D3 region of 28S rRNA gene, and a mtDNA COII region with primer sets 1A/MelR, D2A/D3B, and C2F3/1108, respectively (Power and Harris 1993, Subbotin et al. 2006, Tigano et al. 2005). The sequence of 18S rRNA gene (accession no. MZ948800 haplotype 1 and MZ955998 haplotype 2), haplotype 1 shared 100% identity with that of M. enterolobii from the United States (KP901058) and China (MN832688); haplotype 2 shared 99.8% identity with that of KP901058 and MN832688. The sequence of the D2-D3 region (MZ955995) shared 99% identity with that M. enterolobii from the United States (KP901079). Sequence of the COII region (MZ964625) also shared 99% identity with that of M. enterolobii from the United States (AY446975) and China (MN840970). Phylogenetic trees of the three gene sequences plotted as described by Ye et al. (2021) revealed that the newly described nematode was grouped with M. enterolobii. Sequence analysis of two fragments: 236 bp and 520 bp amplified with gene specific primers Me-F/R and MK7F/R, respectively (Long et al. 2006, Tigano et al. 2010) also confirmed the identity of M. enterolobii. To measure the reproductive factor (Rf), the Poinsettia ‘Luv U Pink’ seedlings with eight true leaves were transplanted into three 12-cm diameter pots each containing 6000 eggs or water (mock control). Forty-five days after inoculation, the average Rf value of three inoculated plants was 6, and no galls were observed on mock control plant roots, confirming that poinsettia is the host of M. enterolobii. M. enterolobii has been reported in several Euphorbia species, including E. heterophylla, E. prostrata, E. punicea and E. tirucalli (Han et al. 2012, Rich et al. 2009). To the best of our knowledge, this is the first report of M. enterolobii infecting E. pulcherrima ‘Luv U Pink’. 

scholarly journals Efficacy of a novel topical combination of esafoxolaner, eprinomectin and praziquantel against fleas in cats, under field conditions

Parasite ◽  
2021 ◽  
Vol 28 ◽  
pp. 22 ◽  
Author(s):  
Eric Tielemans ◽  
Tomoko Otsuki ◽  
Tara Cheesman ◽  
Fiona Selmes ◽  
Anthony Pfefferkorn ◽  
...  
Keyword(s):  
United States ◽  
Field Studies ◽  
The United States ◽  
Positive Control ◽  
Control Group ◽  
The Novel ◽  
Domestic Cats ◽  
Label Dose ◽  
Treatment Tolerance ◽  
Significant Health

Esafoxolaner is a purified afoxolaner enantiomer with insecticidal and acaricidal properties. It is combined with eprinomectin and praziquantel, nematodicidal and cestodicidal compounds, in a novel topical endectoparasiticide formulation for cats. This novel formulation was tested in four field studies, in the United States, Europe, Japan and Australia. In all studies, naturally flea-infested domestic cats were treated with the novel formulation at the label dose and conditions of use. The main objective, identical in the four studies, was to assess efficacy on fleas, based on comparison of mean number of fleas found on infested cats before and one month after treatment. Tolerance to the product was also evaluated in the four studies. Otherwise, the studies had some differences in their design and secondary objectives, for example testing for a reduction in flea infestation-related cutaneous signs, testing of one treatment or of three monthly treatments, and use of a positive control group. In the four studies, a total of 307 cats were treated with the novel formulation. The reduction of fleas one month after treatment was 97.7%, 98.8%, 100% and 99.7% in the United States, Europe, Japan and Australia, respectively. There were no significant health abnormalities attributed to treatment in any of the studies.

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