scholarly journals Comparative Study of VaginalLactobacillus Phages Isolated from Women in the United States and Turkey: Prevalence, Morphology, Host Range, and DNA Homology

2001 ◽  
Vol 8 (1) ◽  
pp. 31-39 ◽  
Author(s):  
Ali O. Kiliç ◽  
Sylvia I. Pavlova ◽  
Sengul Alpay ◽  
S. Sirri Kiliç ◽  
Lin Tao
Keyword(s):  
United States ◽  
Host Range ◽  
Geographic Origin ◽  
The United States ◽  
Rrna Gene ◽  
Broad Host Range ◽  
Dna Homology ◽  
Genetic Types ◽  
Vaginal Lactobacilli ◽  
Reproductive Aged Women

ABSTRACT Lactobacilli play an important role in maintaining vaginal health. However, during bacterial vaginosis lactobacilli decrease for unknown reasons. Our preliminary study showed that phages could infect vaginal lactobacilli. Therefore, the aim of this study was to analyze the distribution, virulence, and types of vaginal Lactobacillusphages isolated from women of two countries: the United States and Turkey. A total of 209 vaginal lactobacilli were isolated from reproductive-aged women in the United States (n = 107) and Turkey (n = 102). By analysis of 16S rRNA gene sequence and by comparison of protein profiles, most lactobacilli were identified as L. crispatus, L. gasseri, andL. jensenii. After mitomycin C induction, 28% of American lactobacilli and 36% of Turkish lactobacilli released phages. A total of 67 phages were isolated and further characterized by their host range, electron microscopy, and DNA homology. All 67 phages were infective against lactobacilli from both collections. The host ranges of most phages were broad, including multiple Lactobacillusspecies. Even though the phages were all temperate, they were able to cause lytic infection in various strains. The electron micrographs of these phages showed a hexagon-shaped head and a long tail with or without a contractile tail sheath. Based on their morphology, these phages belonged to Bradley's phage groups A and B, and could be further classified into four morphotypes. All four types were found among American phages, but only three were found among Turkish isolates. DNA hybridization with labeled probes of the four types of phages revealed that additional genetic types existed within each morphotype among these phages. The phage genomic sizes ranged between 34 and 55 kb. Many of the lysogenic Lactobacillus strains released phages spontaneously at a high frequency of 10−3to 10−4 PFU/cell. In conclusion, lysogeny in vaginal lactobacilli is widely spread. Some lysogenic lactobacilli spontaneously release phages with a broad host range, which can be lytic against other vaginal lactobacilli regardless of their geographic origin.

Lady’s Slipper Orchid and Hydrangea: New Ornamental Hosts of Tobacco Rattle Virus (TRV) in Minnesota

2020 ◽  
Vol 21 (1) ◽  
pp. 19-20
Author(s):  
Mattie M. Baumann ◽  
Roy G. Kiambi ◽  
Benham E. Lockhart
Keyword(s):  
United States ◽  
Host Range ◽  
Tobacco Rattle Virus ◽  
The United States ◽  
Broad Host Range ◽  
Specific Primers ◽  
Virus Particles ◽  
Foliar Symptoms ◽  
Weed Hosts ◽  
Slipper Orchid

The lady’s slipper orchids are a subfamily encompassing over 160 species, including the state flower of Minnesota, Cypripedium reginae. Hydrangea is a genus of about 75 species of shrubs and trees that are popular in perennial gardens. Chlorotic and necrotic foliar symptoms were observed in lady’s slipper orchid and Hydrangea arborescens on plants in St. Paul, Minnesota. From partially purified extracts, virus particles resembling those of tobacco rattle virus (TRV) were observed. TRV-specific primers amplified products from both hydrangea and lady’s slipper and were then sequenced. The sequences matched published TRV sequences with 99% identity, confirming the presence of the virus. TRV has a broad host range including ornamental, vegetable, and weed hosts. This is the first report of TRV infection in both lady’s slipper and hydrangea in Minnesota and the United States.

scholarly journals Porcine Deltacoronaviruses: Origin, Evolution, Cross-Species Transmission and Zoonotic Potential

Pathogens ◽  
2022 ◽  
Vol 11 (1) ◽  
pp. 79
Author(s):  
Fanzhi Kong ◽  
Qiuhong Wang ◽  
Scott P. Kenney ◽  
Kwonil Jung ◽  
Anastasia N. Vlasova ◽  
...  
Keyword(s):  
United States ◽  
Hong Kong ◽  
Host Range ◽  
Acute Diarrhoea ◽  
The United States ◽  
Broad Host Range ◽  
Zoonotic Potential ◽  
Comprehensive Understanding ◽  
Neonatal Piglets ◽  
Turkey Poults

Porcine deltacoronavirus (PDCoV) is an emerging enteropathogenic coronavirus of swine that causes acute diarrhoea, vomiting, dehydration and mortality in seronegative neonatal piglets. PDCoV was first reported in Hong Kong in 2012 and its etiological features were first characterized in the United States in 2014. Currently, PDCoV is a concern due to its broad host range, including humans. Chickens, turkey poults, and gnotobiotic calves can be experimentally infected by PDCoV. Therefore, as discussed in this review, a comprehensive understanding of the origin, evolution, cross-species transmission and zoonotic potential of epidemic PDCoV strains is urgently needed.

scholarly journals Colletotrichum fioriniae infecting invasive Japanese hop (Humulus scandens) in the United States

Plant Disease ◽  
2021 ◽  
Author(s):  
Reid Frederick ◽  
Craig Cavin ◽  
Jami L Thomas ◽  
William L. Bruckart ◽  
Matthew A. Tancos
Keyword(s):  
United States ◽  
Host Range ◽  
Plant Pathogens ◽  
The United States ◽  
Tween 20 ◽  
Broad Host Range ◽  
Post Inoculation ◽  
Native Plant ◽  
Field Samples ◽  
Humulus Scandens

Japanese hop (Humulus scandens) is a non-native, invasive plant that colonizes disturbed riparian areas throughout the eastern United States and Canada, forming dense, monocultural stands that displace native plant communities due to a high reproductive rate, rapid growth, climbing bines, and dense shading (Balogh and Dancza 2008). It is capable of serving as a reservoir for agronomically important plant pathogens, such as the Tomato spotted wilt virus and powdery mildew species that infect commercial hemp and hop fields (Yoon et al. 2018; Weldon et al. 2020). In the spring of 2016, diseased populations of H. scandens were observed along the Monocacy River in Frederick County, Maryland with severe chlorotic and necrotic leaf lesions. Symptomatic leaves were surface sterilized and placed in moist chambers at 25°C for sporulation. Sporulating acervuli, lacking setae, developed on irregular, tan necrotic leaf lesions following 7 to 12 days in a moist chamber (Figure 1). Conidia were hyaline, aseptate, smooth-walled, fusiform to cylindrical with both ends acute (Figure 1B). Conidia measured (n = 100) [L x W; Average (+ Std. Err), range]: 12.42 µm (± 0.10), 8.41 – 14.48 µm; x 3.91 µm (±0.03), 3.03 – 4.91 µm. Monoconidial fungal cultures were obtained by transferring conidia with a sterile glass needle to acidified potato dextrose agar and incubated at 25°C for 2 to 3 days. Based on phenotypic characteristics and conidial morphology and size, the pathogen appeared to belong to the Colletotrichum acutatum complex (Damm et al. 2012). Therefore, six loci (ITS, GADPH, CHS1, HIS3, ACT, and TUB2) were amplified and sequenced from a representative isolate, 16-008, for species characterization (GenBank accessions MW023070 to MW023075) (Damm et al. 2012). For the ITS region and ACT, GADPH, and CHS1 loci, isolate 16-008 was 100% identical to C. fioriniae and shared 99% similarity to TUB2 and HIS3 for multiple accessions of C. fioriniae in GenBank. Gene sequences were aligned, trimmed, concatenated, and analyzed against 32 reference strains, within the C. acutatum complex (Damm et al. 2012). Concatenated loci were used to generate a maximum likelihood phylogeny using W-IQ-TREE (Trifinopoulos et al. 2016). Results from the phylogenetic analysis demonstrated that isolate 16-008 was most genetically similar to C. fioriniae with a bootstrap support of 100% (Figure 2). Based on phenotypic and sequence analyses, isolate 16-008 was identified as C. fioriniae. Humulus scandens seedlings from Maryland (n = 3) were inoculated with a conidia suspension (107 conidia mL-1) with 0.125% Tween 20® and applied with an atomizer until runoff. Inoculated plants were placed in a dew chamber at 25°C for 2 days. Experimental plants were distributed in a mist tent at 25°C with 14 h of light and monitored for 2 weeks. Negative control plants (n = 2) were sprayed with a sterile 0.125% Tween 20® water solution. All inoculated plants were symptomatic by 12 days post inoculation. No symptoms were observed on the mock-inoculated plants. Symptoms were identical to disease field samples. Inoculations were repeated with the same results. Colletotrichum fioriniae was reisolated and confirmed from excised leaf lesions via ITS and ACT sequencing. To our knowledge, this is the first report of C. fioriniae naturally infecting H. scandens within the United States (Farr and Rossman 2020). Future studies will evaluate the host range of this isolate due to the species broad host range and the weed’s extensive distribution.

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 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.

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.

scholarly journals First Report of Stem and Foliage Blight of Chrysanthemum Caused by Phytophthora drechsleri in the United States

Plant Disease ◽  
2021 ◽  
Author(s):  
Charles Krasnow ◽  
Nancy Rechcigl ◽  
Jennifer Olson ◽  
Linus Schmitz ◽  
Steven N. Jeffers
Keyword(s):  
United States ◽  
South Carolina ◽  
Sequence Similarity ◽  
Morphological Characteristics ◽  
The United States ◽  
Universal Primers ◽  
Broad Host Range ◽  
Pathogenicity Test ◽  
First Report ◽  
Foliage Blight

Chrysanthemum (Chrysanthemum × morifolium) plants exhibiting stem and foliage blight were observed in a commercial nursery in eastern Oklahoma in June 2019. Disease symptoms were observed on ~10% of plants during a period of frequent rain and high temperatures (26-36°C). Dark brown lesions girdled the stems of symptomatic plants and leaves were wilted and necrotic. The crown and roots were asymptomatic and not discolored. A species of Phytophthora was consistently isolated from the stems of diseased plants on selective V8 agar (Lamour and Hausbeck 2000). The Phytophthora sp. produced ellipsoid to obpyriform sporangia that were non-papillate and persistent on V8 agar plugs submerged in distilled water for 8 h. Sporangia formed on long sporangiophores and measured 50.5 (45-60) × 29.8 (25-35) µm. Oospores and chlamydospores were not formed by individual isolates. Mycelium growth was present at 35°C. Isolates were tentatively identified as P. drechsleri using morphological characteristics and growth at 35°C (Erwin and Ribeiro 1996). DNA was extracted from mycelium of four isolates, and the internal transcribed spacer (ITS) region was amplified using universal primers ITS 4 and ITS 6. The PCR product was sequenced and a BLASTn search showed 100% sequence similarity to P. drechsleri (GenBank Accession Nos. KJ755118 and GU111625), a common species of Phytophthora that has been observed on ornamental and vegetable crops in the U.S. (Erwin and Ribeiro 1996). The gene sequences for each isolate were deposited in GenBank (accession Nos. MW315961, MW315962, MW315963, and MW315964). These four isolates were paired with known A1 and A2 isolates on super clarified V8 agar (Jeffers 2015), and all four were mating type A1. They also were sensitive to the fungicide mefenoxam at 100 ppm (Olson et al. 2013). To confirm pathogenicity, 4-week-old ‘Brandi Burgundy’ chrysanthemum plants were grown in 10-cm pots containing a peat potting medium. Plants (n = 7) were atomized with 1 ml of zoospore suspension containing 5 × 103 zoospores of each isolate. Control plants received sterile water. Plants were maintained at 100% RH for 24 h and then placed in a protected shade-structure where temperatures ranged from 19-32°C. All plants displayed symptoms of stem and foliage blight in 2-3 days. Symptoms that developed on infected plants were similar to those observed in the nursery. Several inoculated plants died, but stem blight, dieback, and foliar wilt were primarily observed. Disease severity averaged 50-60% on inoculated plants 15 days after inoculation. Control plants did not develop symptoms. The pathogen was consistently isolated from stems of symptomatic plants and verified as P. drechsleri based on morphology. The pathogenicity test was repeated with similar results. P. drechsleri has a broad host range (Erwin and Ribeiro 1996; Farr et al. 2021), including green beans (Phaseolus vulgaris), which are susceptible to seedling blight and pod rot in eastern Oklahoma. Previously, P. drechsleri has been reported on chrysanthemums in Argentina (Frezzi 1950), Pennsylvania (Molnar et al. 2020), and South Carolina (Camacho 2009). Chrysanthemums are widely grown in nurseries in the Midwest and other regions of the USA for local and national markets. This is the first report of P. drechsleri causing stem and foliage blight on chrysanthemum species in the United States. Identifying sources of primary inoculum may be necessary to limit economic loss from P. drechsleri.

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’. 

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