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Dive into cutting-edge science and meet fellow volunteers at our monthly community science event in Malibu
Discover the fascinating world of eDNA water testing
which helps track biodiversity and monitor our Marine Protected Areas
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Although our Malibu eDNA sample sites are accessible
there are many unknown factors that could pose risks to your health and safety
there is limited data on how water quality and sand conditions may have been impacted
making it difficult to determine whether these beaches are safe
our eDNA team has implemented the following precautions:
As community scientists, we trust you to make informed decisions based on the evolving data and understanding of these conditions. Please do not put yourself in any situation where you feel unsafe. If you have any questions or concerns, don’t hesitate to reach out. And for updates as new data become available, check out our Ash to Action page
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A poem can lift the spirits and nourish the soul. This week, let’s all learn one together!
The Poetry Challenge See Full Poem & Readings Play Game
Share full article167167 By A.O. Scott and Aliza Aufrichtig
Welcome to the end of the Poetry Challenge — the last stanza of our five-day adventure in verse. (If you missed the beginning or middle, you can catch up here)
We hope you are feeling reasonably merry and not unduly tired
at an elementary school not far from where Edna St
Students there were invited to write down a poem and carry it around in their pockets
If they ran into somebody who was in urgent need of a poem
You never know when that’s going to happen
We’ve made space in our busy days and our buzzing minds to carry around a bundle of couplets
Some of us may have inflicted “Recuerdo” on other people
But the question nonetheless arises: Now that you have this poem
suggested by our young friend and implied in Millay’s last lines
a souvenir offered by the poet to whomever might have been with her on that ferry ride and
impulsive act of giving that wraps up the night can stand as a metaphor for poetry itself
respond with something that goes far beyond simple charity
a poem acquires meaning only insofar as it is shared
since everyone who acquires it will understand it — will use it — in their own way
Yours to treasure: to recite under your breath
Human memory is a curious thing — variable
Some mental storage spaces are neatly sorted and cross-indexed
Others are more like kitchen drawers overflowing with odds and ends: candle stubs
takeout menus from restaurants that shut down during Covid
Some of “Recuerdo” may start to go blurry — which one of us ate the pear
— but that slippage is anticipated by the poem itself
It’s not a comprehensive accounting of everything that happened
a skein of especially vivid associations and impressions
Your brain may process the poem in a similar way
the words “a bucketful of gold” might pop into your head
The rhymes and alliterations you labored over this week have become part of your verbal muscle memory
And that repeating first couplet — which comprises six of the poem’s 18 lines and occupied the first day of this challenge — will surely jingle in your pocket for a long time to come
poet Watch her full readingWhether you recite the whole thing every day or pick up scraps of it every now and then
That might be as much poetry as you require
Her “Collected Poems” fill more than 700 pages
Or you might prefer her slimmer individual collections
A “Recuerdo” algorithm could just as well point you toward poems about boats or New York
toward lyrics about regretting the dawn or seizing the day
It lives through intuitive leaps and improbable inferences
uncanny connections between common experience and your own imagination
Every poem is its own recommendation engine
and every reader will find a perfectly idiosyncratic way of following it
First, though, let us know how you did. Did you learn it? Would you read it for us? We hope you’ll share your thoughts and suggestions with us below
And we hope that further poetic voyages are in store
“Aunt Jennifer’s Tigers,” by Adrienne Rich
is a blazing portrait of an artist and her work
Scott admires its craft — and its wildness
Scott marvels at the power and paradox of a sonnet by Gwendolyn Brooks
George Oppen’s “From a Photograph” turns a wintry snapshot into a moving meditation on parenthood and the passage of time
is one of the best things that our critic A.O
Scott walks you through a poem that speaks to his mood right now
It’s called “Party Politics,” but it’s not about those parties
Frank O’Hara’s “Having a Coke With You” makes a charming first impression
and right away you want to get to know it better
Share full article167167Edited by Gregory Cowles
Design and development by Umi Syam and Eden Weingart
Additional video production by McKinnon de Kuyper
Metrics details
Surveys of fish diversity in complex heterogeneous environments are highly challenging to perform using traditional survey methods
Although environmental DNA (eDNA) metabarcoding has been effectively used to evaluate fish diversity
studies exploring the spatial and temporal variability of fish communities in mosaic habitats and their connection to water quality after ecological project implementation are still scarce
we evaluated the changes in water quality and fish assembles using the traditional method and environmental DNA (eDNA) metabarcoding after Ecological water replenishment (EWR) and the links between fish communities and water quality were established in the Baiyangdian (BYD) ecosystem in the North China Plain
CODMn and TP showed a conspicuous improvement pattern
and the number of fish species increased notably after EWR
6 more introduced fish species were recorded when compared with the historical data before the implementation of the EWR project
the species richness showed a highly significant difference among the four habitats in the summer 2020 and spring 2021 (spring: P = 0.000; summer: P = 0.002)
and obvious discrimination of fish communities across two seasons was observed (P = 0.001) with eDNA metabarcoding
The water quality variables driving the changes of fish communities during the same period varied significantly across different habitats
while not all showed noticeable discrepancy in driving cross-seasonal fish community changes
our study highlights that the continuous EWR improves the water quality and fish richness but potential ecological issues associated with introduced species should be carefully considered after EWR
Our results also confirm that eDNA is a reliable tool for assessing fish diversity and distinguishing spatiotemporal variability of fish communities in mosaic habitat ecosystems
previous research has predominantly focused on the influence of the EWR on water quality or fish assemblages
limited attention was given to the integrated influence of water quality and aquatic organisms
in which the key factors driving changes in the biotic community remained unidentified
evaluating the changes of water quality and fish assembles after EWR and finding out the key factors affecting the fish communities are of paramount importance for the conservation of fish biodiversity
while numerous natural ecosystems combine multiple habitat types and distributed in interlace
there is an urgent need to address the fish assemblage characteristics using eDNA metabarcoding in mosaic habitat ecosystems
As the recovery of fish community is a long-term process
the changes in fish diversity in BYD ecosystem following the continued EWR (since 2018) warrant further investigation
prior studies have primarily focused on the overall fish community of BYD ecosystem
without clarifying the specific characteristics and differences of fish communities across its interlaced habitats
this study evaluated the changes in environment and fish community structures with the traditional method and eDNA metabarcoding after EWR across the complex heterogeneous habitats
The spatiotemporal distributions of fish communities were linked with water quality factors to finding out the key factors affecting the fish communities
The aims of this study are as follows (i) evaluate the changes in water quality and fish assembles after continuous EWR
(ii) clarify the connection between water quality parameters and fish assemblage dynamics
and identify the primary water quality factors driving spatiotemporal patterns of fish
(iii) assess the ability of eDNA metabarcoding for discriminating the spatiotemporal heterogeneity of fish communities within mosaic habitat systems
The geographical location of the BYD ecosystem (A); a real-world image of four different habitats in the BYD ecosystem (B); distribution map of sampling sites (C)
Five replicate samples were gathered from each sampling location for eDNA metabarcoding
The accession number of samples ranged from SAMN21400475 to SAMN21401107
non-target species that are typically not classified as fish (such as bacteria and viruses) were removed
The species detected by both traditional method and eDNA metabarcoding are considered target species
Target species include fish identified in previous studies or known to exist in adjacent waters
as well as species that were not previously recorded but are capable of surviving in the habitat
Thirteen sampling sites (approximately 20% sites) were selected to conduct parallel sample analysis
CODMn and TN for the five sections from 2018 to 2021 in real time were monitored by fixed water quality automatic monitoring system (LFWAMS-2010
A level of P < 0.05 was used to determine statistical significance upon rejecting the null hypothesis
Species richness of fish detected by eDNA metabarcoding and traditional method in the spring (A)
summer (B) and the two seasons combined (C)
Percentage of phylum richness identified by eDNA metabarcoding (D)
The species richness (A) of four habitats and global scale and relative abundance (B) of shared phyla in spring and summer. The cumulative species richness (C) and relative abundance (D) of four habitats in two seasons. L, T, R, S and G represent the relative abundance of fish at the lake berth, trench, riverway, swamp and global scales.
Relative abundance (%) of fish per habitat at the species level (A, B) and order level (C, D) with eDNA metabarcoding. Habitats codes were consistent with those in Fig. 3
Nonmetric multidimensional scaling (NMDS) ordination of the entire dataset for four habitats in spring (A)
the entire dataset for four habitats in summer (B) and the global scale in two seasons (C)
RDA plots of the fish communities and water quality variables in spring (D)
temporal trends in water quality variables after EWR were analyzed from 2018 to 2021 in the BYD ecosystem
the fish community status was obtained with eDNA metabarcoding and traditional method
spatiotemporal dynamics of fish communities were evaluated using eDNA metabarcoding
and the relationship between fish assemblages and water quality variables was analyzed within mosaic habitats from BYD ecosystem
Our findings indicated that all water quality variables exhibited a noticeable improvement pattern
and the fish richness was richer than historical data in the BYD ecosystem
the fish communities differed significantly across multiple habitats and seasons
and the spatiotemporal variations in fish communities were closely linked to water quality variables in the BYD ecosystem
nobilis posed a serious threat to native planktivorous fish in the Mississippi River basin
our findings indicate that the water quality improved and the species richness increased but the potential ecological problems of the introduced species should be of concern after EWR in the BYD ecosystem
our study further shows that the water quality factors driving the changes of fish communities in complex habitats of BYD ecosystem during the same period varied significantly across different habitats
and nutrients were the main driving factors for the distribution of fish communities
our results demonstrate that the water quality variables driving the changes of fish communities during the same period varied significantly across different habitats
and that the relative abundance of fish species detected exclusively by eDNA metabarcoding was generally low in the Rhône River basin
our findings further indicated that eDNA metabarcoding is a promising tool for monitoring fish diversity by effectively detecting low-abundance species and promoting more realistic census of the fish community
further showed that eDNA metabarcoding was capable of effectively differentiating fish assemblages at stream and river locations in French Guiana
this study suggested the fish communities differed significantly across multiple habitats and seasons in BYD ecosystem
and eDNA metabarcoding is useful for detecting spatiotemporal heterogeneity of fish assemblage in ecosystems with distribution of different habitats in interlace
The changes in the content of water quality variables and species richness of fish were evaluated and analyzed after EWR in the BYD ecosystem
the spatiotemporal dynamics of fish assemblages and interactive networks between communities and water quality variables were analyzed using eDNA metabarcoding
Our results highlighted that the water quality improved and the species richness increased but the potential ecological problems of the introduced species should be of concern after EWR in the BYD ecosystem
this study further shows that the water quality factors driving the changes of fish communities in complex habitats during the same period focus on the variables with significant differences across different habitats
while which not all for the cross-season fish community for the differences in water quality requirements for fish across seasons
our finding suggested eDNA metabarcoding can be utilized to assess temporal-spatial dynamics of the fish assemblages in mosaic habitat ecosystems and discriminate the spatiotemporal heterogeneity of fish communities
to effectively restore and conserve native fish resources
long-term monitoring of fish communities and water quality factors following continuous EWR is strongly recommended
Raw sequence reads are deposited in the NCBI Sequence Read Archive database BioProject ID: PRJNA565582 (https://www.ncbi.nlm.nih.gov/sra/PRJNA734376)
Assessment of fish communities using environmental DNA: Effect of Spatial sampling design in Lentic systems of different sizes
Human impacts on global freshwater fish biodiversity
Fish conservation in freshwater and marine realms: Status
Fish biodiversity conservation and restoration
urgently needs ‘scientific’ and ‘ecological’ action
Research on variation of water quality before and after ecological water supplementation in the Beijing section of the Yongding river
Water chemical characteristics and water quality evaluation of the river under the ecological water replenishment: A case study in the Yongding river basin in North China
Effects of water replenishment on lake water quality and trophic status: An 11-year study in cold and arid regions
An ecologically dispatch strategy using environmental flows for a cascade multi-sluice system: A case study of the Yongjiang river basin
Changes in fish community structure in the old channel of Tianezhou in the Yangtze river from 2017 to 2021
genetic analysis and enlightenment for ex situ conservation of the Yangtze finless porpoise
Hydraulic assessment of environmental flow regimes to facilitate fish passage through natural riffles: Shoalhaven river below Tallowa dam
eDNA metabarcoding revealed differential structures of aquatic communities in a dynamic freshwater ecosystem shaped by habitat heterogeneity
Present situation of fish stocks in Baiyangdian lake
Comparison of environmental DNA metabarcoding and conventional fish survey methods in a river system
eDNA metabarcoding as a promising conservation tool for monitoring fish diversity in a coastal wetland of the Pearl river estuary compared to bottom trawling
Environmental DNA metabarcoding uncovers environmental correlates of fish communities in spatially heterogeneous freshwater habitats
Environmental DNA metabarcoding for biodiversity monitoring of a highly diverse tropical fish community in a coral reef lagoon: Estimation of species richness and detection of habitat segregation
Environmental DNA survey captures patterns of fish and invertebrate diversity across a tropical seascape
Analysis on the potential of EWR for Baiyangdian Lake by reservoirs in the mountainous area of Daqing River
& Seasonal variation of aquatic macrophytes and its relationship with environmental factors in Baiyangdian lake
Present condition analysis of Hebei Baiyangdian lake fish stocks after diversion from yellow river to the lake
7–2014; Technical Guidelines for Biodiversity Monitoring—Inland Water Fish
Ministry of Ecology and Environment of the People’s Republic of China (Beijing
Seasonal and habitat influences on fish communities within the lower Yasuni river basin of the Ecuadorian Amazon
Quantification of mesocosm fish and amphibian species diversity via environmental DNA metabarcoding
FLASH: Fast length adjustment of short reads to improve genome assemblies
Modeling the ecological status response of rivers to multiple stressors using machine learning: A comparison of environmental DNA metabarcoding and morphological data
UPARSE: Highly accurate OTU sequences from microbial amplicon reads
Spatial variation in macrobenthic assemblages and their relationship with environmental factors in the upstream and midstream regions of the Heihe river basin
Water quality assessment and variation trends analysis of the min river sea-entry section
Estimating aquatic plant diversity and distribution in rivers from Jingjinji region
using environmental DNA metabarcoding and a traditional survey method
Inter-season and interannual variations in fish and macrocrustacean community structure on an Eastern english channel sandy beach: Influence of environmental factors
Comparison of environmental DNA metabarcoding and a traditional survey method for assessing fish diversity and distribution along salinity gradient in an urban brackish reservoir
Assessment of benthic invertebrate diversity and river ecological status along an urbanized gradient using environmental DNA metabarcoding and a traditional survey method
Simultaneous assessment of the Macrobiome and Microbiome in a bulk sample of tropical arthropods through DNA meta systematics
Picante: R tools for integrating phylogenies and ecology
On a test of whether one of two random variables is stochastically larger than the other
Fish community structure and its relationship with environmental factors in Baiyangdian lake
How do the variations of water and sediment fluxes into the estuary influence the ecosystem
Species diversity: From global decreases to local increases
Quantifying the effects of introduced Bighead carp (Cyprinidae; Aristichthys nobilis) stocking on dominant fish species in the Ulungur lake
Mesohabitat associations in the Mississippi river basin: A long-term study on the catch rates and physical habitat associations of juvenile silver carp and two native planktivores
Isotope niche dimension and trophic overlap between bigheaded carps and native filter-feeding fish in the lower Missouri river
Fish assemblages respond to altered flow regimes via ecological filtering of life history strategies
Effects of resource availability and hydrological regime on autochthonous and allochthonous carbon in the food web of a large cross-border river (China)
Northern rivers ecosystem initiative: Nutrients and dissolved oxygen–issues and impacts
Carassius cuvieri,·Red Carassius Auratus) (Shanghai Science & Technical
Fish community structure in relation to acidity in three Nova Scotia rivers
Fish assemblage in different types of irrigation ditches in Huixian wetland
Seasonal dynamics of riverine fish communities using eDNA
Uncovering the complete biodiversity structure in Spatial networks: The example of riverine systems
Spatial and temporal distribution of fish and zooplankton in a shallow lake
Composition of fish communities in German lakes as related to lake morphology
Navigation impacts on freshwater fish assemblages: The ecological relevance of swimming performance
Habitat-specific fishing revealed distinct indicator species in German lowland lake fish communities
Unlocking biodiversity and conservation studies in high-diversity environments using environmental DNA (eDNA): A test with Guianese freshwater fishes
Download references
This work was supported by the National Key Research and Development Project (2019YFC1803402) and National Natural Science Foundation of China (21976202)
and Xinyuan Shi for assisting in sample collection
National Engineering Research Center of Industrial Wastewater Detoxication and Resource Recovery Research Center for Eco-Environmental Sciences
Engineering Research Center of Green development for Conventional Aquatic Biological Industry in the Yangtze River Economic Belt
State Key Laboratory on Environmental Aquatic Chemistry
Research Center for Eco- Environmental Sciences
visualization and writing - original draft
Wenqiang Zhang: investigation and bioinformatics data analysis
Saihong Yan: investigation and bioinformatics data analysis
writing - review & editing and project supervision
The authors declare no competing interests
This study was conducted in strict accordance with the laws governing animal experimentation in China
The protocols were approved by the the Ethics Committee for Animal Experiments of Jimei University and Research Center for Eco-Environmental Sciences
All efforts were made to minimize suffering
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations
Below is the link to the electronic supplementary material
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DOI: https://doi.org/10.1038/s41598-025-99970-1
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Edna was a member of Grace Bible Church in Berne
She was a lifelong homemaker and retired from Troyer’s Market following 18 years of service
She was extremely hardworking and loved hosting people
she cherished the time spent with her family and friends
Indiana; and Carol Schwartz of Shipshewana
Indiana; 23 grandchildren; and six great-grandchildren
she was preceded in death by four brothers
A funeral service for Edna will be held at 10:00 a.m
Burial will follow at Westlawn Cemetery in Geneva
The family will receive friends from 12 noon to 3:00 p.m
Preferred memorials can be given to Galilean Home Ministries
Arrangements are by Zwick & Jahn Funeral Homes
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great-grandmother and great-great-grandmother Phyllis Balfour passed away on Saturday
at home surrounded by her loving son and family members
to Samuel Albert Sudbury and Bertha Hedwig (Kunkel) Sudbury
She married the love of her life Donald Balfour on February 15
Together they raised four boys: Donald Balfour
She is survived by her son Darrin and her four daughters-in-law Judy
nine great-grandchildren and twelve great-great-grandchildren
Phyllis was preceded in death by her husband Donald and her three sons Donald
and sisters Marie (Sudbury) Jenson and Irene (Sudbury) Callahan
Phyllis was a committed homemaker and relished making her home
patio and garden a welcoming gathering spot for family and friends
She enjoyed dressing up in all her beautiful colorful dresses and going out to dinner with family and friends
She loved looking back on the camping memories she made with her husband
She enjoyed many years of season tickets to Hale Center Theater (playhouses) on a regular basis
Phyllis supported Donald in his business venture of Balfour Plumbing where she was a bookkeeper for over 50 years
which has now turned to a 3 generation company
She left a legacy of the importance of family
want to thank the amazing hospice team at Active Home Health
Hospice & Personal Care for all their amazing work in keeping Phyllis happy and healthy till the age of 99
We also want to give our deepest appreciation to a few visitors who came to see Phyllis on a weekly basis
A great thanks to Chris for his final blessing
Phyllis will be laid to rest next to her husband Don at Utah Veterans Memorial Park on June 27
A celebration of life is planned for early June
quietly passed Tuesday April 29 in Lawrenceburg IN
1927 in Lexington to Charlie and Mary Cordelia Fightmaster Carr
she was retired from IBM and a lifelong member of Old Union Christian Church where she sang in the choir and taught Sunday School
She was a member of the Rosie Reds for many years and a die-hard UK basketball fan
painting and most of all her family and friends
grandchildren Jessica (Kevin) Milburn of Greenville KY
Anna (Joey) Finley of Bowling Green KY and Zachary Britton of Lawrenceburg IN
Funeral services will be 12:30 Monday May 5
Burial to follow in Evergreen Memory Gardens
Visitation will be 10:30 – 12:30 prior to service
In lieu of flowers contributions may be sent to Old Union Christian Church Choir 6856 Russell Cave Rd
Edna Earle McCormick Cheek passed away peacefully at home on Tuesday
South Carolina to the late Wilburn Earl and Bessie Moore McCormick
She graduated from McColl-Fletcher Memorial High School in 1958
Edna worked as a teacher’s aide for the Richmond County school system
and later for the Winston-Salem/Forsyth County school system
She enjoyed spending time at her and Marshall’s second home on Oak Island
Edna also enjoyed attending University of South Carolina football games with her family for many years and continued to follow the Gamecocks when she was no longer able to attend in person
she was a talented seamstress; having made her evening gown when she was crowned Miss McColl High as well as making a wedding gown for her sister
The family has many of her treasured creations to remember her by
Edna was preceded in death by her parents; and by Marshall
and granddaughter Mallory of Wilmington; daughter Amy
grandson Thomas and granddaughter Emma of Kernersville; sister Edith Lewis of North Myrtle Beach
S.C.; sister Beth Stone and niece Jessie Stone of Columbia
SC; nephew Kiah Stone and family of Charleston
the family asks for donations to Trellis Supportive Care at 101 Hospice Ln
The family will receive friends on Saturday
from 10:00-10:45 AM at Hayworth-Miller Kernersville Chapel
followed by a memorial service at 11:00 AM in the funeral home
Kernersville Chapel - Hayworth-Miller Funeral Homes & Crematory
views and stories from the front lines of conservation
Editor’s note: From “blue carbon” to “ecosystem services,” environmental jargon is everywhere. Conservation International looks to make sense of it in an occasional explainer series we’re calling “What on Earth?"
a wildlife monitoring tool that is changing the way we study the natural world
I keep seeing this thing called "eDNA" in science articles
animals are constantly leaving tiny traces of themselves in their environment — in the water
These microscopic breadcrumbs contain bits of genetic material: DNA
Scientists call this “environmental DNA,” or eDNA for short. By collecting samples from a pond, a river or possibly even a cloud of dust
researchers can analyze the DNA fragments left behind to figure out what species have been there — even if they never saw them
A stream in the Alto Mayo Protected Forest where scientists took samples to test for eDNA
which helps them rapidly detect species that are difficult to observe directly
scientists don’t need to spot an animal to know that it’s there
A scientist can scoop up a bit of pond water
and figure out which critters have been hanging out there
“eDNA is great for finding species that are rare
secretive or live in places people can’t easily reach,” said Ali Swanson
who leads nature technology at Conservation International
“The technique works especially well in water
where DNA easily mixes and moves around — a single sample can give us a surprisingly rich snapshot of the broader watershed
including areas that would be tough to survey in person.”
What kinds of animals can you find with eDNA
a notoriously difficult species to spot in the wild
eDNA breaks down pretty quickly in the environment — usually within days — so it’s great for detecting recent or current presence
no Jurassic Park — but it does sound like it can really help protect wildlife.
eDNA can provide solid scientific evidence that can help attract funding for conservation or inspire new protections for threatened ecosystems
That discovery is now helping to shape a new ecological corridor that will connect Alto Mayo to other protected areas across the region — giving wildlife more room to roam
So does this mean eDNA can replace other methods
It’s great at telling you what species are present — but it’s still hard to say how many individuals there are
acoustic sensors and good old-fashioned fieldwork still play an important role
“Acoustic monitoring is great for finding birds or insects
camera traps are often ideal for larger mammals
which excels at uncovering shy or rare critters
is part of a mosaic of tools that together reveal a more complete picture of an ecosystem.”
this "blob-headed" fish was discovered on a recent research expedition in the Alto Mayo Protected Forest
When can I investigate my own back yard using eDNA
most eDNA tools are still lab-based and require some serious gear and know-how
But scientists are working on making the process faster
you’re already part of a movement that’s crowdsourcing science
Will McCarry is a staff writer at Conservation International. Want to read more stories like this? Sign up for email updates. Also, please consider supporting our critical work
Edna was a member of Southeast Church of the Nazarene and attended Kent Church of the Nazarene
She was very active in the Nazarene Mission Society and went on Mission trips to several countries
She helped organize and build the Stow Church of the Nazarene and taught Bible Study to hearing impaired students
Edna enjoyed reading and cake decorating and will be deeply missed by all who knew and loved her
she was preceded in death by her husband Lynn Fulper in 1981; brothers
Bernard and Earl Scott and her sister Lena Douthitt
2025 at 12:00PM at Bissler and Sons Funeral Home
Entombment will follow at Grandview Memorial Park Mausoleum
Visitation will be prior to the service from 11:00AM-12:00PM on Friday at the funeral home
please visit the Tribute Wall at www.bisslerandsons.com
Edna Bennett Pierce Prevention Research Center
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The family of Edna Pearl Watt created this Life Tributes page to make it easy to share your memories
You can reach us 24/7/365 at
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ExpandJoliet City Councilwoman Jan Hallums Quillman
Edna Keith Elementary School Principal Laticha Walton
Joliet Public Schools District 86 Deputy Superintendent Dr
Joliet Public Schools District 86 Assistant Superintendent Dr
City of Joliet Tree Advisory Board Member Betsy Satcher
and Tree Advisory Board Chairperson Rita Renwick pose by the new Summer Elixir Elm tree at Edna Keith Elementary School on April 25
(Photo Provided by Joliet Public Schools District 86)
Joliet — Edna Keth Elementary School hosted this year’s annual Joliet Arbor Day celebration by planting a new tree in front of the school
Joliet Mayor Terry D’Arcy attended the event, along with City Council members Jan Hallums Quillman and Pat Mudron
and members of the city’s Tree Advisory Board
D’Arcy read an official Arbor Day proclamation
and Edna Keith kindergarten students recited a poem before the school’s winners of the city’s annual student Arbor Day poster contest were announced
Fifth grader Eian McMillan took first place in the poster contest
followed by third grader Ja’Mya McMurtry in second place and second grader Nestor Tinoco Rodriguez in third place
Fourth grader Annabelle Cordova and fifth grader Jackson Hollister also were recognized as honorable mentions
The students were recognized at the Joliet City Council meeting earlier in April.
Edna Keith Elementary School Arbor Day poster contest winners Eian McMillan, Ja’Mya McMurtry, Nestor Tinoco Rodriguez and Annabelle Cordova pose for a photo April 10, 2025. (Photo Provided by Joliet )
Students from the school, located at 400 4th Ave., then were able to assist the city officials, school Principal Laticha Walton, and Joliet Public Schools District 86 Deputy Superintendent Tanisha Cannon and Assistant Superintendent Judie Nash in planting a young summer elixir elm tree in front of the building.
The city donated the tree as well as a new book for the school’s library, along with Midwest crabapple tree saplings for each of the 72 kindergarten students to take home and plant.
“We were pleased to be selected to host the city’s Arbor Day event,” Walton said. “The students were thrilled to be part of the celebration.”
The city of Joliet was recognized as a “Tree City USA” community for the 34th consecutive year by the National Arbor Day Foundation.
Copyright © 2023 Shaw Local News Network
Share full article184184 By A.O. Scott and Aliza Aufrichtig
Yesterday we put our heads down to look closely at Edna St. Vincent Millay’s “Recuerdo” — at the words and syllables and even individual letters that make it a poem. (If you’re just joining us, this is the third day of our Poetry Challenge. There’s plenty of time to catch up, but you may want to start on Day 1.)
Those words are about something; they come from somewhere; they belong to someone
we find ourselves in New York City in one of its legendary eras
in the company of one of its great characters
“Recuerdo” is a quintessential New York poem
not only because it happens to be about a popular form of public transportation between two of the city’s island boroughs
This all-night boat ride is a great urban adventure
And perhaps only Millay could have turned it into literature
Millay was a fixture of the city’s literary and political bohemia
counting many of its best-known artists and intellectuals among her friends and lovers
eminently quotable snippets of rhymed gossip — pulse with the dynamism and attitude of the modern city
taking in sights and sounds that are still ubiquitous more than a century later
novelist Watch her full readingNobody has time for the daily newspaper
actor Watch his full readingExhaustion nips at the heels of delight; the ferries and subways never stop running
in 1892 and grew up mostly in nearby Camden
in what a biographer called “the smallest house in the poorest part” of that seaside mill town
whose mother couldn’t afford to send her to college
attracted the attention of a wealthy patron who raised the money to send her to Vassar
Millay became notorious on campus for her love affairs with men and women
her brilliance and her disregard for the rules
she published her first book of poems and moved to Greenwich Village
streaking like a comet through Manhattan’s creative scene
“A Few Figs From Thistles” (1920) became a touchstone of its era
The four lines of “First Fig” catch the heedless
headlong spirit of New York in the Jazz Age:
It can be hard for a 21st-century reader to grasp just how famous these poems made her
winning prizes and garnering appreciative reviews in little magazines
She was a young phenom who grew into a wildly popular artist and a durable celebrity
At some point around the end of World War I
she may have spent a night riding the Staten Island Ferry
possibly in the company of the Nicaraguan poet Salomón de la Selva
But whether or not “Recuerdo” is based on a real-life event
it breathes the air of reality — the sounds
smells and sights of a city with the unique power to tire you out and fill you with joy
poet Watch her full readingThe poem is in good company
beckoning poets of the past and future to join it on deck
Millay’s criss-crossing of the harbor invokes Walt Whitman
whose “Crossing Brooklyn Ferry,” published in 1856
On the ferry-boats the hundreds and hundreds that cross
And you that shall cross from shore to shore years hence are more to me
Walt Whitman, “Crossing Brooklyn Ferry”
Whitman is talking to Millay’s future nocturnal passengers, and to the rhymers and dreamers who would populate the metropolis in centuries to come. To Hart Crane, who hymned “the harp and altar” of the Brooklyn Bridge. To Audre Lorde, who wrote about
Audre Lorde, “A Trip on the Staten Island Ferry”
There’s a great New York poem for every light on Broadway. Another one is Sara Teasdale’s “Broadway.”
This night is ours, of all the golden nights,
“This night is ours” may be the perfect distillation of New York as a poetic state of mind — an anticipation of “Recuerdo.” The city that never sleeps is an experience to be shared.
In today’s game, we’ve translated that experience into pictures. See if you can put it back into words!
“Aunt Jennifer’s Tigers,” by Adrienne Rich, is a blazing portrait of an artist and her work. Our critic A.O. Scott admires its craft — and its wildness.
Our critic A.O. Scott marvels at the power and paradox of a sonnet by Gwendolyn Brooks.
George Oppen’s “From a Photograph” turns a wintry snapshot into a moving meditation on parenthood and the passage of time. Our critic A.O. Scott shows you what he loves about it.
“Romantic Poet,” by Diane Seuss, is one of the best things that our critic A.O. Scott read (and reread) this year.
Our critic A.O. Scott walks you through a poem that speaks to his mood right now. It’s called “Party Politics,” but it’s not about those parties, or those politics.
Frank O’Hara’s “Having a Coke With You” makes a charming first impression, and right away you want to get to know it better.
Share full article184184Edited by Gregory Cowles, Alicia DeSantis, Nick Donofrio and Joumana Khatib. Additional editing by Emily Eakin, Tina Jordan, Laura Thompson and Emma Lumeij. Design and development by Umi Syam and Eden Weingart. Additional design by Victoria Pandeirada. Video production by Caroline Kim. Additional video production by McKinnon de Kuyper. Photo editing by Erica Ackerberg. Illustration art direction by Tala Safie. Illustrations by Hannah Robinson.
Share full article639639 By A.O. Scott and Aliza Aufrichtig
Someone once defined poetry as “memorable speech.” By that standard
each of us has committed at least some poetry to memory
song lyrics and movie catchphrases all find their way into our heads
More formal memorization used to be a common classroom ritual
Schoolchildren would stand and recite approved works for their teachers and peers
That kind of learning has mostly gone out of fashion
which may be a sign of progress or a symptom of decline
school shouldn’t be the only place for poetry
And learning a poem by heart doesn’t have to be drudgery
It can be a way of holding onto something beautiful
a morsel of verbal pleasure you can take out whenever you want
A poem recited under your breath or in your head can soothe your nerves
drive away the noise of everyday life or grant a moment of simple happiness
less commodified corner of your consciousness
It’s a flower in the windowbox of your mind
You could store a whole anthology in your brain
But let’s start with one: “Recuerdo,” by Edna St
“Recuerdo,” first published in 1919 in Poetry magazine
is the recollection of a night out on the town — or more precisely on the water
presumably the stretch of New York Harbor served by the Staten Island Ferry
We asked some friends of the Book Review — poets
actors and other literature lovers — to recite it for us
Ina Garten and Ethan Hawke will introduce you to the poem
poet laureate Watch her full readingWhy did we pick “Recuerdo”
We combed through our shelves like Goldilocks
looking for a poem that was just right: not too difficult
but not too simple; not obscure but not a chestnut; not a downer but not frivolous either
and we thought something that rhymed would be more fun — and easier — to memorize than a cascade of free verse
who was born in Maine in 1892 and was a fixture of the Greenwich Village bohemian scene in the 1920s
John Lofman/Published with permission of The Edna St
She was a decidedly modern author who often wrote in traditional forms
and who has stayed popular through 100 years of fluctuating fashion
When you get to the second stanza of “Recuerdo,” read here by Ina Garten
cook and author Watch her full readingIt’s a city poem
but one that incorporates some arresting nature imagery (the sun
It delivers a confidential message — addressed to a “you” who shares the memory of those moments by the fire and in the moonlight — while striking a convivial
The poem concludes with an impulsive act of generosity that carries a hint of melancholy
The night is over; another day is here with its obligations and routines; we’re about to trade the open air of the ferry for the crowded underground platforms of the subway
though of course there’s more to it than that
The poem expresses the desire to hold on to a fleeting experience
to fix it in words and images before it’s washed away on the tide of time
The speaker summons bits and pieces of a memorable night
organizing them into verses that bring those hours back to life
and — impossibly but also unmistakably — we’re right there with her
So here is the challenge: Memorize this poem
you’ll find a game designed to help you learn “Recuerdo.” Today your goal is to master that wonderful refrain
you’ll have one third of the poem.) As the Challenge continues through the week
we’ll look closely at how the poem is made
at what it’s about and at the extraordinary woman who wrote it
There will be new games and videos every day
Share full article639639Edited by Gregory Cowles
Edna Faye Newman (Riffey) of Knoxville TN passed on to her Heavenly home on March 17
2025 due to complications of Heart and Kidney failure
She took every opportunity she could to talk about the Lord
She was a member of Roseberry Baptist Church
She loved singing in the choir and was a member for many years
She was a 1960 graduate of Central High School
playing golf and especially mowing her own yard
She enjoyed throwing parties on holidays for her family
Mom always talked about what a wonderful family and extended family she was blessed with
She was preceded in death by her loving husband of 57 years
Jerry D Newman; her parents Dallas and Christina Riffey Sr.; her parents in laws Clarence and Willie Mae Newman
Most of her large family was waiting on her arrival
and Jack Riffey(Ginny); Sisters Flora Etta Leadbetter,(John) Frances Stalsworth.(Tommy)
Betty Lou Irwin(Kenneth); and Brother-in-law
Lisa Thomas (Anthony); her granddaughter Ashley Thomas and boyfriend Jay Goins; and son
Jessie Riffey and Glenda Newman; Brothers-in-law
Also leaving behind numerous nieces and nephews
She had special friends in Lisa Chambers-Dodson and Kenny Dodson
They were always there when she needed them
The family will receive family and friends on Thursday
2025 from 5:00 PM to 7:00 PM with a service to follow at 7:00 PM at Mynatt Funeral Home Fountain City Chapel
The family asks that in light of her memory to please wear flashy jewelry just like Edna would
Mynatt Funeral Home is honored to serve the family of Edna Newman and condolences may be expressed at www.mynattfh.com
passed away with her family by her side on Monday
a daughter to the late Adam Washington and Carrie (McKinney) Black
she also earned a bachelor’s degree in theology from Adullam Bible College in Jackson
family and education was important and essential to a happy productive life
Edna would say to her children “give your life to Christ
Early in life Edna worked various jobs in the community to make sure all her children were well taken care of and she selflessly brought children in need into her home and worked as a foster mother for over 15 years
She never saw a problem to big that prayer and her faith in God could not overcome
Her powerful prayers reached heaven that pulled her son Mark through a life threating incident at the age of 9
Her children as well as the community relied on her anointed prayer life to help them through difficult times
a loving mother and married to the late A.C
She was a devout Christian and church member
serving as a prominent church mother and missionary of the House of Restoration and Grace Christian Center both in Worcester
MA as well as New Life Christian Center in Jackson
Edna received the 2015 Impact award as Mother of the year from New Life Christian Center under the leadership of Apostle Barry Chaney
Edna was not only a prominent church mother
but she was also a mother to every community she touched.
Edna also had the privilege of being lovingly embraced by the families of her dear friends
District Missionary Jacqueline Coppedge of Worcester
Whenever Missionary Coppedge would visit Edna
Elder Frank and Missy Mary Jardine of Worcester
MA who she would make it her business to have dinners ready for them to take home after Sunday’s church service
Edna’s doors and heart were always open which afforded her the privilege of entering their hearts and home
providing her with extended families that deeply cherished her.
sewing and hat making as all her daughters and daughter in-laws
and granddaughters are wearing to celebrate her memory and some of her monumental designs
Her passion was delivering the gospel of Jesus Christ to everyone who would listen
Edna was wise in her delivery of winning souls for the Kingdom of God
she would always have a kind word of encouragement
One of Edna’s favorite scriptures was Psalms 118:17-18 KJV (I shall not die
but live and declare the works of the Lord)
such as “I’m A Soldier in the Army of the Lord”
She would always say I love to hear my Apostle Chaney sing “I love You Lord”.
and Clarence Estell (Charmaine) of Oklahoma City
Janice Rose (Pastor Alexander) of Worcester
Edna was graced with twenty eight (28) grandchildren
fifty one (51) great grandchildren and one (1) great-great grandchild; three (3) foster sons
Tommy Collins and Kevin Jones and their families
Carl Vester McKinney and Rosco McKinney; a sister
Dorothy McKinney; a host of nieces and nephews the entire McKinney family Shirley
Extended children: Jimmy Carruthers of Osceola
Relatives and friends are invited to a period of calling hours on Tuesday
from 10-11 am immediately followed by her funeral service at 11:00 am in Grace Christian Center
Worcester is honored to be assisting the family with arrangements
Pastor Alexander Rose (House of Restoration) and Entire Church Family
Pastor Emma Black (Grace Christian Center) and Entire Church Family
Chaney and Co Pastor Angelia Chaney (New Life Christian Center) Entire Church Family
Bishop Everett and Minister Kateena Gates family (One In Christ) and Entire Church Family of Clermont
and First Lady Jacqueline Webbs (Mt Sinai COGIC) and entire church family
Bishop Bernard and First Lady Sandra Reese (Framingham COGIC) and entire church family
Elder Benjamin and Lucy Marshall of Framingham
Mother Dora Blue and the late Jimmy Blue of Natick
Edna Mae GriffinBirth date: May 1
Share a story where Edna's kindness touched your heart
Describe a day with Edna you’ll never forget
Edna C Cervantes, a beloved wife, mother, and grandmother passed away on Friday March 21, 2025, at the age of 70 years old. Edna was born on March 24, 1954, to her late parents Frank and Gloria Cerda, and lived her entire life in San Antonio,... View Obituary & Service Information
Cervantes created this Life Tributes page to make it easy to share your memories
© 2025 Mission Park Funeral Chapels & Cemeteries
Made with love by funeralOne
passed away peacefully at home on December 22
She was the youngest and only girl in a household of three older brothers
Her father frequently told her that she was “his favorite daughter.”
Teddy grew up in the Avenues area of Salt Lake City and later moved to a home bordering the Bonneville Golf Course
She attended East High School and the University of Utah where she affiliated with the Chi Omega Sorority along with some of her closest and lifelong friends
Teddy graduated from the University of Utah with two bachelor's degrees
She loved spending the summers at her family cabin in upper Weber Canyon
the entire ranch became a playground and fostered close lifelong relationships
Her cousins became as close as brothers and sisters
As a mother she passed onto her children her love of the Weber and surrounding mountains
She also enjoyed boating and spending time at the beach that often included a stay with her brother Chick
Teddy loved gardening and would call it her “daily exercise” as she spent hours on her hands and knees tending to her yard into her nineties
Teddy was sealed for time and all eternity to James Grant Stringham in the Salt Lake Temple on August 10
Their marriage continued for sixty-three years until Grant’s death in 2017
Teddy and Grant initially lived in Baltimore Maryland while Grant attended medical school at the University of Maryland
She taught elementary school in Baltimore to help support their family
She continued teaching school when they moved to Illinois
They returned to Salt Lake City and established their permanent home in the Parley’s Third Ward where they lived for the next sixty-two years
Mom was an accomplished musician and pianist
Having received formal training in piano and organ
she often accompanied vocal performers and found time to teach piano to each of her children and others
Teddy had a deep love and testimony of the Savior which motivated her lifelong commitment to service of others
She was a devoted member of The Church of Jesus Christ of Latter-day Saints
She served in many capacities of leadership including Ward and Stake Relief Society President
She also served as ward pianist and organist until well into her eighties
She was involved in her community and was always one to provide service to others
Some of her favorite community positions included serving as President of the Medical Auxiliary
and as delegate to the Utah Republican State Convention
She worked with the students of Edison Elementary in Salt Lake City for the next twenty years
she began tutoring and supportive programs to help students with educational assistance and basic necessities
Teddy especially loved working at Edison because of the impact she could make in her students’ lives
Our remarkable mother had an indominable and enduring spirit
but tempered her strong will with love and kindness
Her constant love and dedication to her family was keenly felt and will be sorely missed
Her family was the single most important thing in her life
She constantly strived to maintain close relationships with each of them
She is also survived by 12 grandchildren and 14 great-grandchildren
Teddy was preceded in death by her husband Grant
and her great-grandson Christopher Groscost
The family would like to extend heartfelt appreciation to Amanda Laloni
and other care givers for their loving service that allowed Teddy to remain in her home during the final years of her life
2025 at 12:00 pm (noon) at the Parleys Third Ward
2615 Stringham Avenue (2295 South) Salt Lake City
Friends may call Saturday morning at 11:00 am prior to the funeral
Interment will be in Salt Lake City Cemetery
You may watch a recording of the service online through Zoom by clicking the “Watch Service” button above
Always made me feel so special in your heart
I’ve got birthday voicemail saved from you which I’ve listened to when I’m feeling down
My parents Connie and Jerry thought so highly of her
Living in the same neighborhood for 46 years was so wonderful
Teddy was my mother's friend since kindergarten and I saw her in a couple of the college photos
A Celebration of Life and Memorial Service is to be held at Concordia Lutheran Church in Jamestown on February 22 at 3 pm
Interment will be in the Kensal Cemetery at a later date
to Edward and Rosine (Wahl) Rosenau in Jamestown
She graduated from Jamestown High School in 1947 after which she attended Valley City Teachers College and received her Teaching certificate
She worked as a teacher in a one room schoolhouse until she married Leonard Norheim on June 10
Leonard and Edna lived on the family farmstead and raised four children northwest of Kensal
Paul Lutheran Church of Kensal serving on the Auxiliary and as a Sunday school teacher
One of her fondest memories is that she was a member of the Eldridge basketball team
She also enjoyed her employment at Pipestem Creek where she crafted natural bird feeder arrangements
Edna’s hobbies included watching MN Twins baseball
as well as attending local basketball games with friends
She was preceded in death by her parents; husband Leonard; daughter Sandra Donat; brothers
Harlan and Ray; and two nephews and one niece
one of six children born to the late Arthur and Dora Shaw Dixon
Edna was the owner and operator of Hill House Sportswear in Cana
She is survived by her beloved husband of 59 years
Mildred Dixon Bartlett; several nieces and nephews
and Lara Dixon Payne; several great-nieces and great-nephews; and a sweet exchange daughter
Catherine Dixon Richardson; and three brothers
at 4:00 PM at Moody Funeral Home Chapel in Mount Airy
The family will receive friends Monday from 3:00 until 4:00 PM at the funeral home
memorial contributions may be made to the Tunnel to Towers Foundation
NY 10306; or Mountain Valley Hospice and Palliative Care
all deaths handled by Crain Funeral Home & Cremation Service must have a minimum of the name uploaded to our website
Made with love by funeralOne
Edna Geraldine Silvey-Ross (nee Wilcox) of O’Fallon
passed away peacefully into the arms of her savior on January 9
Edna was the devoted wife of both the late Edgar Silvey and Lester Ross
Edna is the cherished daughter of the late Joe B and Ida Grace Wilcox
and preceded in death by her siblings Robert
Chantel (Brian) Summers; great-grandchildren
Skyler and Thomas Summers and great-great-grandchildren
and she cherished every moment spent with them.
Edna entered the workforce after being widowed in her younger years and served as both mother and father in a time when that wasn't common
Edna served as a telephone operator at Southwestern Bell for over 30 years before transitioning to a position at JC Penney after her retirement.
tending to her plants and flowers and spending time outdoors in the fresh air regardless of the weather
She loved the water even though she never learned to swim
Her warm spirit and love for her family will forever be remembered
and devotion to her family and friends will live on in the hearts of all who knew her
A celebration of her life will be held from 4-7:00 pm on Tuesday
A funeral mass will be held at 10:30am on Wednesday
at Assumption Catholic Church in O'Fallon
She will be laid to rest immediately following the mass at Calvary Cemetery in St
On Saturday, March 1, 2025, Edna “Chocolate” Thomas Morgan transitioned to her heavenly home at the age of 90. She was a beloved mother, family member, educator, friend, and servant of the Lord. She was blessed with a long, fulfilled... View Obituary & Service Information
Edna Thomas Morgan created this Life Tributes page to make it easy to share your memories
Available by phone 24/7
Copyright © 2025 Taylor Funeral Home of Chattanooga, Inc.. All Rights Reserved.
Share full article165165 By A.O. Scott and Aliza Aufrichtig
If you’re joining us in memorizing Edna St. Vincent Millay’s “Recuerdo” this week, you probably already have the first two lines stuck in your head. (If you’re just discovering the Poetry Challenge, please check out yesterday’s introduction
filmmaker Watch his full readingOnce you’ve got these
since this repeating couplet functions as a mini-chorus at the start of each stanza
That refrain tells the story in a nutshell
But this poem is more than just a report on one night on the ferry
It recreates the voyage through a flurry of sensory details
Those features — the imagery and the sound; what your mind’s eye sees and your physical ears hear — are what make “Recuerdo” a poem
and paying attention to how they work can help us learn it
the poem captures how it felt to be on that boat
You can see the sky turning color as the morning air breezes up
poet Watch her full readingYou can taste the fruits of the voyage
novelist Watch her full readingStrain your ears just a little
and you can make out the sounds of boats in the harbor
novelist Watch his full readingThese impressions — and the vividness of Millay’s language — can help anchor the poem in your mind
But the secret to fixing it in your memory is to learn its structure
to listen to the musical patterns of its language
and many of its features originated as aids to memory in an oral
It’s easier to find the word you’re looking for if you know it sounds like the other words around it
alliteration and rhythm are not only pleasing to the ear; they’re sticky
Each line of “Recuerdo” is a poetic wave that breaks on the shore of a rhyme
poet Watch his full readingRhyme is just one of the ways poets use repeating sounds to make their work memorable
and the English language has a fondness for it that goes back to its earliest literature
clusters of consonants in the middle of the lines knot them together and help you hold on to them
playwright and novelist Watch his full readingThe poem’s individual words and syllables bob like a string of harbor buoys
Every line is propelled by the cadence of stressed and unstressed syllables
novelist Watch her full readingThis pattern of rhythm and sound — four-beat lines yoked in rhyming pairs — is a familiar one in English
You may have encountered it before you could read
Every Who down in Who-ville liked Christmas a lot
Joni Mitchell, “Both Sides Now”
Andrew Marvell, “To His Coy Mistress”
there’s a lot of variation within the basic pattern — longer or shorter lines
but it’s also intuitive and physical — it lives in the bobbing of your head or tapping of your foot as you read
and while most English poems (including “Recuerdo”) have varying feet
Millay often places her strong beats after two unstressed syllables: da-da-DUM
novelist Watch her full readingIn others they spatter like raindrops:
TV host and noted book lover Watch her full readingWords are more than sounds and syllables
The words in “Recuerdo” form a bouquet of arresting images and sensations
an experience that will be different for each reader
we have pulled apart some of the components of the poem
you can’t really separate sound from sense
and work together to create something that resists summary
What does a bucketful of gold look like to you
What face do you see when the shawl-covered head turns to acknowledge your greeting
See how many of its words you already have
Share full article165165Edited by Gregory Cowles
Metrics details
Environmental DNA (eDNA) is revolutionizing how we investigate biodiversity in aquatic and terrestrial environments
It is increasingly used for detecting rare and invasive species
assessing biodiversity loss and monitoring fish communities
as it is considered a cost-effective and noninvasive approach
but samples collected from these settings may exhibit PCR inhibition and a low fish read recovery
Here we present an approach for detecting fish in turbid
The workflow includes bead-based extraction
high fidelity and specificity DNA polymerase (Platinum SuperFi II) and multiplexing the universal MiFish primers
By applying this hybrid method to a variety of complex estuarine samples with known inhibition
we have more than doubled the number of recovered fish species while removing most of the off-target amplification
we have developed an eDNA workflow with two main protocols
optimized for detecting fish in turbid estuarine samples
We evaluated each step in these protocols to identify a workflow that is adaptable to robotic platforms
and optimizes the number of fish species and the number of generated fish Amplicon Sequence Variants (ASVs)
Our final selected workflow included an automated KingFisher DNA extraction system
which utilizes magnetic beads for high-throughput DNA isolation
This protocol incorporates the Zymo OneStep™ PCR Inhibitor Removal Kit
to reduce inhibitors commonly present in environmental samples
recognized for its high fidelity and specificity in amplifying low-concentration DNA
which progressively lowers the annealing temperature during the initial cycles to enhance specificity and amplification efficiency
This approach is particularly effective when Multiplexing Mifish primers
which are designed to detect a broad range of fish species
The protocols in this workflow were tested on 48 samples collected from four estuaries in the United States’ National Estuarine Research Reserve System (NERRS) a nationwide network of protected estuarine areas dedicated to research
and stewardship to inform coastal management
This geographical diversity in site selection was intended to ensure a broad spectrum of environmental conditions
Despite reductions in sediment supply due to damming and sediment retention upstream
turbidity remains a defining feature of the estuary
The NERRS System-Wide Monitoring Program (SWMP) collects long-term water quality data at all NERR sites
including this one (see Table S2 in the Supplementary Information)
Elevated turbidity at this site presents challenges for eDNA sampling
including filter clogging and PCR inhibition caused by organic and inorganic particulates
the estuary sampling sites encompasses a substantial portion of the salinity gradient
ranging from salt marsh conditions at China Camp to brackish marsh at Rush Ranch
This estuary also experiences large salinity gradients due to variable freshwater inflows from the Mission and Aransas rivers
low river flows and high evaporation can lead to hypersaline conditions in shallow bays
while freshwater pulses from rainfall events can significantly lower salinity levels for extended periods
This variability influences nutrient dynamics and biological productivity
resulting in fluctuations of organic matter concentrations
humid subtropical climate with significant seasonal temperature variations
The estuary receives substantial freshwater input from the Apalachicola River
creating dynamic salinity gradients influenced by seasonal river flows
and tidal exchanges with the Gulf of Mexico
This variability leads to fluctuations in nutrient levels and high biological productivity
supporting a rich biodiversity that includes important commercial fisheries and rare species
The varying freshwater inflows result in changes in turbidity and organic matter concentrations
The reserve features temperate conditions with well-defined seasons and experiences significant seasonal temperature variations
The Mullica River-Great Bay Estuary within the reserve is characterized by low-gradient
southeast-flowing streams that originate from groundwater inflows
These streams have high concentrations of humic acids and organic matter from decaying vegetation
leading to brown-colored waters rich in dissolved organic carbon
Seasonal salinity gradients occur due to variable freshwater inputs
we can test our eDNA workflow across a range of challenging environmental conditions
This approach ensures that the protocols we developed are robust and adaptable for detecting fish eDNA in various turbid estuarine environments
we tested two DNA extraction methods to evaluate their effectiveness for fish eDNA detection in estuarine environments: the KingFisher automated bead-based extraction and the Qiagen-based extraction
optimized for high-throughput applications
uses paramagnetic beads to automate and streamline DNA isolation
while the Qiagen-based extraction relies on a silica column
a trusted method widely used in eDNA studies
We compared these two approaches to determine which method delivers the highest DNA yield and quality
particularly when working with challenging turbid samples
to improve the overall efficiency and reliability of eDNA-based fish species detection
As interest in molecular monitoring increases
the use of robotic systems in eDNA extraction and handling is becoming more common
especially for processing larger numbers of samples
Robotic systems offer advantages such as increased efficiency and consistency
though they also have potential disadvantages
including the risk of cross-contamination and higher initial costs
Our goal was to validate an automated bead-based extraction method with comparable performance to commonly used column-based methods like QIAGEN kit-based extractions
Samples were extracted with the automated Kingfisher protocol using magnetic beads
and DNA concentrations ranged between 1.84 ng/μl and 25.8 ng/μl (Fig
we observed variability among the sites with Mission-Aransas displaying the highest yields (Fig
Variability within each site reflects conditions at each of the four sampling locations (Table S1
We did not observe any cross-contamination effects
and we found no significant differences between DNA concentrations from samples obtained with the bead-based Kingfisher extraction and the QIAGEN protocol (p-value = 0.7; Fig
This suggests that our automated DNA extraction using magnetic beads is equally effective under various conditions
PCR inhibition removal using Zymo OneStep
Panel 1a: Samples from the San Francisco NERRS site show no detectable PCR amplification due to inhibition when using the KAPA protocol without inhibition removal (KAPA.CTRL)
Panel 1b: The introduction of the Zymo OneStep™ PCR Inhibitor Removal Kit (KAPA.ZYMO) results in visible amplification but also generates off-target DNA fragments
as indicated by additional bands above the expected range
Panel 1c: Using the Zymo kit in combination with the Platinum SuperFi II polymerase and a touchdown PCR program
and the resulting DNA fragments fall within the expected range of 200–300 bp
M: DNA ladder (marker) indicates the fragment size
Inhibition is prevalent across many estuarine samples processed in our lab
often muting or completely preventing PCR amplification
Laboratory managers might opt to apply inhibition removal selectively based on initial PCR results or incorporate it routinely into the workflow to consistently improve outcomes
The Zymo OneStep™ PCR Inhibitor Removal Kit costs approximately $2.07 per sample and using Platinum SuperFi II PCR Master Mix adds around $2.78 per sample
bringing the total increase to $4.85 per sample excluding labor costs
These steps significantly improve the specificity and success of amplification
that an overly specific polymerase will result in non-detection of some target species
was generated using data from three separate E-gels
as indicated by the boundaries visible in the figure
The absence of a single full-length gel image is due to the original experimental setup involving separate gel runs
The impact of inhibition removal on ASV numbers was minimal (mean of 13 ASVs for KAPA.CTRL and 14 with KAPA.ZYMO); however, the comprehensive integration of Zymo with the Platinum protocol markedly increased the mean number of ASVs per sample to 46 (Fig. 2b)
Enhanced fish detection across individual Sites
Box plots comparing the distribution of fish species between the three tested protocols at four different estuaries (lower panel map)
each location has different environmental conditions and expected species
the use of platinum with Zymo cleanup resulted in a higher detection rate of fish species at every site
Statistical significance (p < 0.05) between treatments was assessed using Tukey HSD tests
Platinum.ZYMO was significantly different from both KAPA.CTRL and KAPA.ZYMO
KAPA.ZYMO and Platinum.ZYMO were significantly different from KAPA.CTRL
all three treatments showed significant differences from one another
The Mish-E primer was added in response to feedback from resource managers that elasmobranchs (rays and sharks) were not being detected in areas where they were known to be present
Although we did detect skate (family Rajidae) with the MiFish-U alone
the combined primers detected four additional elasmobranchii belonging to three different families: Dasyatidae (Dasyatidae spp.
Comparison Between Fish Reads and Raw reads
A linear regression analysis shows an increase in sequencing depth correlates with an increased detection of fish ASVs but not species
The gray shaded area represents the 95% Confidence Interval
Diversity of Fish ASVs Across San Francisco Bay
Maximum Likelihood (ML) trees depicting the diversity of unique fish ASVs detected across three different treatments: KAPA.CTRL (control without inhibitor removal)
and Platinum.ZYMO (with Platinum SuperFi II polymerase and Zymo inhibitor removal)
which are color-coded based on their taxonomic fish group
The trees are built using the ASVs detected from San Francisco Bay samples
with each treatment showcasing different ASV richness and distribution
an outer ring displays bar plots of log10-transformed read counts
where each bar represents the abundance of a specific ASV
Bar colors correspond to the fish groups of the ASVs
visually illustrating the variation in read abundance and taxonomic representation between treatments
The Platinum.ZYMO protocol shows the highest diversity and read abundance of fish ASVs
reflecting the enhanced sensitivity and accuracy of this protocol in detecting fish eDNA in complex environments like San Francisco Bay
or PCR artifacts such as errors or stochastic variations in sample processing
Applying a cutoff threshold of 1% of fish reads to remove rare ASVs resulted in 26 ASVs for KAPA.ZYMO assigned to 19 species and 25 ASVs for Platinum.ZYMO assigned to 20 species
represented only 40 reads for KAPA.ZYMO but 3,010 reads for Platinum.ZYMO
By setting a minimum cutoff of 50 reads per ASV for Platinum.ZYMO
we successfully recovered all 25 fish species and the majority of ASVs (223)
This demonstrates the method’s robustness in detecting species while managing potential artifacts
which is an algorithm for resolving true biological sequences from noisy data
enhance the reliability of detected ASVs by removing errors and chimeras
thereby reflecting true biological variation
Recent updates to platforms like MitoFish and MiFish Pipeline have expanded reference datasets
improving species identification for many taxa
These limitations underscore the need for careful interpretation of rare ASVs to avoid distorting community composition analyses
Bioinformatic processing using denoising methods
and establishing appropriate cutoff thresholds remain critical steps in validating the authenticity of rare ASVs
highlight the value of ASVs in providing higher taxonomic resolution and accuracy
especially when appropriate filtering steps are applied
These studies indicate that while OTUs can artificially inflate diversity by including spurious low-abundance sequences
ASVs offer a more accurate and reliable depiction of community composition
provided that denoising and abundance filtering are rigorously applied
bacterial sequences) distorting species detection
along with filtering and denoising approaches
can mitigate these effects by focusing on high-quality
thereby improving species-level identification and quantifying eDNA abundance with higher precision
while OTUs have traditionally been used in fish eDNA metabarcoding
the growing body of evidence supports a shift toward ASV-based approaches
ASVs not only provide more accurate species-level identifications but also offer greater insights into genetic diversity
which are essential for monitoring biodiversity in dynamic ecosystems like estuaries
we enhance the reliability and resolution of species detection
ultimately contributing to more effective conservation and management strategies
supporting data robustness and reducing the likelihood of spurious results due to PCR variability (Fig
Enhanced fish detection in stream waters
Box plots comparing the distribution of fish ASVs and species between the control (KAPA.CTRL) and the new protocol (Platinum.ZYMO) on stream water samples
Significant p-values are indicated by asterisks with *** for p < 0.001
significantly enhances target fish DNA amplification while minimizing off-target reads
This reduction in non-target amplification is particularly important for estuarine systems
where high levels of bacterial and organic matter often complicate eDNA analysis
In addition to Platinum SuperFi II polymerase
and a dual strategy of DNA dilution combined with an inhibitor removal kit (Zymo OneStep™)
which gradually reduces the annealing temperature
improved the specificity of amplification and reduced non-specific amplifications
allowing for more efficient targeting of low-concentration fish DNA in the presence of inhibitors
the combination of DNA dilution with the Zymo inhibitor removal kit was instrumental in mitigating the effects of PCR inhibitors that are prevalent in highly turbid environments
This combined approach not only prevented PCR inhibition but also optimized amplification efficiency
reducing interference from environmental contaminants and improving the quality of detected fish DNA
The use of both inhibitor removal and touchdown PCR in our workflow increased species detection by over 25% compared to traditional methods
This improvement underscores the effectiveness of integrating advanced techniques—such as inhibitor removal
and optimized PCR strategies—into eDNA workflows for challenging environments
Our findings highlight the importance of refining each step of the eDNA process
to enhance the sensitivity and reliability of fish species detection
particularly in complex and inhibitor-rich environments like estuaries
To validate our workflow, we tested the automated DNA extraction and the three PCR protocols in Table 1 on samples collected from four estuaries
Forty-eight samples were collected during the same period (May 2023) from four different locations within each estuary (12 from each site) to ensure that seasonal or time-related variables do not confound across-site comparisons
We then reviewed the results to evaluate the steps that provide the greatest benefit in terms of number of fish species and fish ASVs
we use the term species to refer to both cases where ASVs were matched to the species level and instances where ASVs could not be matched to the species level but were assigned to a higher taxonomic rank
All taxonomic assignments are based on a lowest common ancestor approach
where sequences are matched to the most specific taxonomic level possible using the available reference database
The samples used in this study were collected from a range of NERRS sites (see above) as part of a pilot project incorporating eDNA into a long-term multi-site monitoring network
then transported to a nearby lab for filtering
Each sample was filtered through 1.5um porosity glass fiber filters and filters were replaced if clogging occurred
The filters were stored in pre-prepared tubes containing 4 ml Longmire’s buffer (Longmire et al
Up to 3 filters from the sample were included in one tube
All lab work was conducted in dedicated Biosafety Level 2 (BSL2) molecular laboratory facilities at the University of New Hampshire
All steps from sample extraction through sequencing were performed in separate
We adhered strictly to standard laboratory cleaning practices for BSL2 including the separation of Pre- and Post-PCR laboratory spaces
PCR Mix preparation was done in one designated area
while DNA template addition was performed in a physically separate space to minimize the risk of contamination
All experiments were conducted under laminar flow hoods and Biosafety cabinets for sample preparation
10% bleach and dH2O and were exposed to UV light before and after experimentation
Qiagen-based extraction: Filters were placed in a lyse and spin basket with 400 ml of buffer ATL and 20 μl of proteinase K and incubated at 56 ◦C for one hour
The remainder of the filter extraction was performed on a QIAcube Connect system (QIAGEN®
Germany) following the QIAamp DNA Mini protocol (Qiagen Cat
This kit is designed for extraction of DNA from tissue and is readily adapted to the QIAcube automated system
Although the reagents are proprietary the manufacturer notes that the method removes inhibition and contaminants
Samples from both methods were eluted to 100 μl
DNA concentration was determined using the Qubit dsDNA HS Assay kit (Thermo Fisher
consisting of H2O and plain buffer without filter
50 μl of the sample was diluted 1:5 and stored in a -20C freezer for use in this project
The remaining sample was archived in a -80C freezer for potential future use
ZYMO OneStep-96 PCR Inhibitor Removal Kit (CAT: D6035) was used to remove inhibition from all samples following the manufacturer’s instructions
150 µl of Prep Solution were added to each Silicon-A™-HRC Plate well
and then centrifuged at 3,500 × g for 5 min
50 µl of DNA was added to the prepared plate
The filtered DNA was used for subsequent PCR reactions
Initially we tested a variety of published PCR protocols as well as several polymerases and inhibitors on samples from these sites. We then selected the most promising alternatives for more detailed testing. The different PCR workflows we evaluated are summarized in Table 1 below
All the PCR comparisons were performed on samples extracted using the automated Kingfisher extraction method
and enhance PCR efficiency and reliability in the presence of inhibitors
The overall goal of this work is to improve eDNA-based fish species detection from problematic turbid samples collected in estuarine environments
PCR Verification Amplification of target regions was verified on 2% E-Gel electrophoresis (Thermo Fisher
Absent or muted bands in samples with verified DNA concentrations indicate that the samples are likely to contain inhibiting agents
E-Gel images are also used as an initial screen for successful amplification of positive controls
and absence of contamination in negative controls
The value of PCR replicates on fish species detection
was tested with the optimized protocol incorporating Platinum and Zymo clean up
Samples from each of the four sites were tested
These replicates were then sequenced as separate samples
Our initial protocol used KAPA HiFi HotStart (KAPA) and a touchdown (TD) thermal profile modified from Pitz et al
The cycling conditions were the same as described for Platinum above except that the annealing temperature of the second cycling phase was set to 55 °C
The PCR reaction was established in a total volume of 12 μl
incorporating 6 μl of KAPA Master Mix (2X)
0.7 μl of the forward and 0.7 μl of the reverse MiFish-U primers
Both types are important in estuarine systems
and multiplexing these primers expands the list of potential target species
The primer and adapter sequences used in this study are listed below
Primer & adapter sequences used in this study
MiFish-U-F: 5′- TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGGTCGGTAAAACTCGTGCCAGC-3′
MiFish-E–F: 5′- TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGGTTGGTAAATCTCGTGCCAGC-3′
MiFish-U-R: 5′- GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG CATAGTGGGGTATCTAATCCCAGTTTG-3′
MiFish-E-R: 5′- GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-CATAGTGGGGTATCTAATCCTAGTTTG-3′
PCR products were submitted to the Hubbard Center for Genome Studies (HCGS) for library preparation and sequencing
the amplicon products from the 1st PCR were prepared by incorporating dual-index barcodes and Illumina sequencing adapters (P5 and P7) into the DNA fragments
The PCR amplification conditions were set as follows: initial denaturation at 94 °C for 3 min
followed by 15 cycles of denaturation at 94 °C for 20 s
A final extension phase was conducted at 72 °C for 7 min to ensure complete amplification of target sequences
The amplified libraries were purified using BluePippin to selectively remove short fragments and primer dimers
We ran feature classification at default parameters except we set the number of accepted reference sequences (percent-identity) to 90% (default is 80%)
and the number of accepted sequences to ‘all’ instead of the top 10 (maxaccepts)
The BLAST search results were retained for confirming classifications for each ASV
“fish reads” refer to the total number of sequencing reads assigned to fish species after the raw data are processed and filtered
Fish reads provide a measure of how much DNA from fish species is present in the sample
fish ASVs represent unique genetic sequences identified in the sample
These ASVs allow us to differentiate between individual species and
The trees were generated under the General Time Reversible (GTR) model with a gamma distribution to accommodate rate variation across sites
This modeling choice is particularly effective for sequences exhibiting high variability
as is common with environmental DNA samples from diverse fish populations
A one-way Analysis of Variance (ANOVA) was performed separately for Fish species and Fish ASVs using the `aov` function in R to determine if the observed differences using our optimized protocol were statistically significant from the other treatments
The null hypothesis for each ANOVA was that the means of Fish species and Fish ASVs for all treatment groups were equal.The primary objective of the statistical analysis was to evaluate the effects of different treatments on both Fish species and Fish ASVs
The treatments included were KAPA.CTRL (control)
The p-values for the treatment comparisons were extracted from the ANOVA results
indicating significant differences among the treatment groups
a post-hoc Tukey’s Honest Significant Difference (HSD) test was conducted using the ‘TukeyHSD’ function in R to identify which specific treatment groups differed from each other
All statistical analyses were conducted using R software (version 4.3.2) with the ‘aov’ function for ANOVA and the ‘TukeyHSD’ function for post-hoc comparisons
To further validate our optimized protocol on data collected from stream waters in New England
1000 ml of water per sample were filtered using a 0.45 µm Whatman cellulose nitrate filters and were processed in the same way for DNA extraction and PCR as our four NERRS sites
For these samples we used a T-test to determine if there was a statistically significant difference between the control treatment "KAPA.CTRL" and the optimized protocol "Platinum.ZYMO"
we conducted a linear regression analysis to evaluate the relationship between sequencing depth (total raw reads) and the detection of fish ASVs and species
This analysis was performed using the lm function in R (version 4.3.2)
with fish ASVs and species regressed against the total number of raw reads
To estimate the precision of the regression line
a 95% Confidence Interval (CI) was calculated using the predict() function in R
and the confidence interval was visualized with the ggplot2 package
This approach allowed us to assess whether increased sequencing depth is associated with improved detection of fish ASVs or species
The raw datasets used and analyzed in this study are available on the NCBI Sequence Read Archive (SRA) under the project number PRJNA1136877. Supplementary data (Version v3), including ASVs and comprehensive lists of detected fish species, can be accessed on Zenodo at https://doi.org/https://doi.org/10.5281/zenodo.12753119
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Bioinformatic analyses were supported by New Hampshire- INBRE through an Institutional Development Award (IDeA)
from the National Institute of General Medical Sciences of the NIH
Department of Civil & Environmental Engineering
designed the methodology and led the writing of the manuscript; F.E.B conducted the experiments and optimization steps
and W.K.T conducted the initial DNA extraction optimization
All authors contributed critically to the drafts and gave their final approval for publication
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DOI: https://doi.org/10.1038/s41598-025-85176-y
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finally made her escape at 7:25am on February 3
as the sun was rising up over the Mississippi near Canton
1926 to Victor and Imogene Dieterich Howell
She graduated from Wyaconda High School in the spring of 1944
After her beloved Leland returned home from the front lines of Europe
1945 and were blessed with over 74 years together!
Edna gratefully spent nearly all of her life in Clark County
Other than during the war after high school and working in Ft
she worked well into her 80’s in Clark County at Moore Forms
Survivors include sons Max (Judith) of Waxahachie
Felix and Rex; and many nieces and nephews.
She was also preceded in death by her close friends Eula
On those rare occasions when Edna would travel
She was renowned for her pie crusts and the world’s greatest cinnamon rolls
was a talented gardener and a loyal friend
loved trail riding horses and being on the farm in Chambersburg
The Schuster’s are especially thankful for the love and affection showered on mom and dad over the years by Jerry and the crew at Wiss & Wiss
We are also thankful for the care and support mom received these last few years as a resident at the Lewis County Nursing Home.
Graveside services for Edna Schuster will be held at 1:00 pm on Saturday
A visitation with family present will be held on Saturday
from 12:00 pm to 12:45 pm at the Wilson & Triplett Funeral Home in Kahoka
Memorials contributions in memory of Edna Schuster are suggested to the Clark County Senior Center
You are invited to share your memories of Edna and leave a condolence at www.wilsontriplett.com
Metrics details
Invasive snakes are among the most challenging invaders worldwide due to their exceptionally low detection rate and grave ecological impacts
Environmental DNA (eDNA) has emerged as a promising tool to improve invasive snake detection and enhancing management programs
yet its application to terrestrial snakes remains underexplored
This study provides the first advances in the use of eDNA techniques to detect the terrestrial invasive California kingsnake (Lampropeltis californiae)
californiae-specific primers and tested their effectiveness in detecting the species in different environmental samples
including swab samples from underneath artificial cover objects (ACOs) made of different materials
we conducted a controlled experiment to assess the accumulation and degradation rate of L
californiae eDNA over a 14-day period (7 with snakes in the terraria and 7 after having removed them)
while no detections appeared in randomly collected soil or controls
eDNA was undetectable in terraria prior snake introductions
but remained detectable throughout the study
with no evidence of snake eDNA degradation after snake removal
These findings provide key insights for the implementation of an eDNA-based protocol for the detection of L
offering a valuable tool for monitoring this invasive species
this study could be used for refining eDNA methodologies to detect other elusive terrestrial snake species elsewhere
the high probability of the species being introduced in continental Europe has led to its inclusion in the List of Invasive Alien Species of European Concern (Regulation (EU) 2022/1203)
which makes it also a conservation priority at the European level
the primary objectives of this study were to (1) design and validate specific primers for L
(2) evaluate various sampling methods for collecting and detecting L
californiae eDNA accumulation and degradation to inform detection protocols
The sampling and laboratory protocols developed could be useful and informative for the implementation of eDNA techniques in the detection of other terrestrial snakes elsewhere
as well as two individuals of each endemic reptile species (own samples
available in our facilities at the CSIC (Tenerife
californiae using the commercial kit E.Z.N.A.® Tissue DNA (Omega Bio-tek Inc.
whereas we used ZR Genomic DNA Tissue MicroPrep Kit (Zymo Research
USA) to extract DNA from the endemic reptiles
We assessed DNA quantity and quality using a NanoDrop®ND-1000 spectrophotometer (Thermo Fisher Scientific
species-specific sequence that distinguished L
We then tested primer specificity by amplifying previously extracted DNA from L
californiae and the endemic reptiles using the same PCR conditions as previously described
with an annealing temperature of 57 °C and the subsequent visualization of PCR products via 1.7% agarose gel electrophoresis stained with Real-Safe (Real Biotech Corporation
we amplified extracted DNA using quantitative PCR (qPCR) with 10–20 ng of DNA
and 7.5 µL of iTaq Universal SYBR Green Supermix (Bio-Rad
We performed reactions on an iCycler BIO-RAD real-time thermal cycler at the Genomic Service of the University of La Laguna
In order to optimize amplification conditions
we first tested a gradient of temperatures and cycle numbers using L
californiae DNA as the positive control and a sample without DNA as the negative control
The final qPCR conditions comprised an initial denaturation at 95 °C for 10 min
followed by 40 amplification cycles (15 s at 95 °C
We assessed primer specificity by comparing qPCR melting curves
californiae-specific primers produced distinct melting curves
while non-specific amplification resulted in spurious curves for endemic reptiles
Map of the study area showing the location of our sampling sites and artificial cover objects (ACOs)
We visited ACOs to collect swab samples following the patterns described in the top inset
we used new disposable latex gloves to collect each sample
stored all swab samples in individual paper envelopes
We incorporated silica gel beads into the swab and boot bags with the objective of mitigating potential mold growth
we consolidated the small individual plastic bags into larger hermetic bags for each sample type and site
No contact with the target species had occurred prior to or during the sampling day
since all sampling and personal material (in addition to the researchers) travelled to Gran Canaria from the uninvaded island of Tenerife for each sampling session
To reduce subjectivity in the interpretation of melt curves
and only samples with clearly similar melting curves and Ct values to those of L
californiae were considered as positive (we repeated those that were ambiguous)
To minimize contamination during laboratory procedures
we handled DNA isolates and prepared qPCR reactions using filtered pipet tips and under the laminar flow hood in the pre-PCR laboratory
We performed all tasks in a dedicated working area within the pre-PCR laboratory
which was thoroughly cleaned with 10% diluted bleach after processing each sample
We used new disposable gloves to manipulate each sample and to access to the freezer
and all instruments were sterilized with 10% diluted bleach
rinsed with distilled water and dried with disposable paper towels
We carried out amplifications in a post-PCR laboratory within the CSIC facilities that did not share equipment or material with the pre-PCR laboratory
We transferred samples between pre- and post-PCR laboratories in a single direction (pre-PCR to post-PCR)
and we spaced the pre-PCR and post-PRC work at least 24 h apart
All substrate sample collection were carried out with care to prevent direct contact with visible urine or feces
Potential sample contamination was minimized by using new disposable latex gloves for each sample and storing them in a − 20 °C freezer located outside the facilities where terraria were situated and where no samples of L
We transported all samples to the CSIC laboratory facilities for subsequent analysis
We extracted the DNA in the CSIC laboratories using a self-designed protocol with the following steps: firstly
we transferred the contents of each sample tube into a new 2 mL tube
we rubbed a sterile cotton swab moistened with ultrapure water (Ultra Pure Water System
USA) over the walls of the empty tube; thirdly
we cut the tip of the swab and dropped it into a 1.5 mL tube
This protocol reduced the time of sample processing without compromising the sensitivity of the technique (tested against other two alternative sample processing methods; see results of those other methods in Supplementary Information II)
We extracted DNA from the swab tips using the commercial extraction kit E.Z.N.A.® Tissue DNA (Omega
All procedures were approved by the Government of the Canary Islands under permit no
Specificity of the self-designed primers for the cytochrome oxidase I subunit
Panel A shows the amplification products (≈ 161 bp) obtained from the extracted DNA of Lampropeltis californiae
Chalcides sexlineatus and Tarentola boettgeri and visualized through 1.7% agarose gel electrophoresis with Real-Safe (Real Biotech Corporation
Taiwan) (the third-last and last lanes represent the negative controls (-C) and the second-last lane included the 25–700 bp DNA marker (LADD-DN1-500; OXGEN™)
Panel B shows the melting temperature curves obtained through qPCR when amplifying the extracted DNA from tails of the four species
Sampling site 2 accounted for 75.00% of all detections
with sampling sites 1 and 3 producing 8.33% and 16.67% of all snake detections
25.00% of which had one positive qPCR replicate
12.50% had two positive replicates and 62.50% had all three positive replicates
62.50% of all detections occurred in corrugated steel plates
The spatial distribution of snake detections during the second period exhibited a similarity to that of the first period
with 50.00% of all detections occurring in sampling site 2
We did not detect positive samples in January
whereas February and March exhibited the highest percentage of positive detections (20.00% and 13.33%
respectively) (detection rate ≤ 10.00% for the remaining months)
We did not find significant differences in eDNA detection rate between periods for swab samples (\(\chi_{1}^{2}=\) 0.01
we observed a significant variation in the detection rate among sites (\(\chi_{2}^{2}=\) 13.07
P = 0.001) and types of materials (\(\chi_{4}^{2}=\) 25.39
Site 2 had a substantially higher detection rate compared to the other sites (standardized residual of 3.46
while site 3 demonstrated a significantly lower rate (standardized residual of -2.65
Corrugated steel plates showed a higher detection rate compared to the remaining materials (standardized residual of 3.98
and used to analyze eDNA degradation over time after Lampropeltis californiae individuals were removed from terraria
This persistent failure underscores an urgent need to explore and adopt novel methodologies to enhance management efficacy
The purpose of this research was to initiate the study of the use of eDNA techniques in the detection of L
an objective that aims to guide the control given the elusive behaviour of the species
we designed and tested primers for the COI subunit
Given the relative paucity of development in eDNA sampling protocols for terrestrial environments in comparison to the aquatic counterparts
our comparative analysis on the probability of detecting L
californiae eDNA from swab samples from ACOs made of five different materials
suggested that swab samples under ACOs represent the most effective sampling protocol currently available (particularly effective with corrugated steel plates and corrugated bitumen sheets)
we demonstrated that the detection rate remains consistent irrespective of the intensification of the sampling regime or the number of ACOs
the detection rate might be enhanced if the sampling is conducted during the months of February and March
and during the seven days after an individual L
californiae was in direct contact with the substrate
In order to implement the use of eDNA techniques in the routine management of L
further work is needed to optimise detection protocols (including refining primer specificity or eDNA amplification protocols)
sampling techniques (including further research on the type of samples to be collected in the field and their location to increase detection rate)
as well as to deepen the knowledge on the species eDNA accumulation and degradation in natural environments
The future development of these techniques to infer L
californiae density could be of great use for conservation practitioners
The datasets generated during and/or analysed during the current study are available at Figshare and have the following DOI number: https://doi.org/10.6084/m9.figshare.28202660.v1
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Gallo (GESPLAN S.A.) for their support to our research
González for providing laboratory materials and equipment from the Biochemistry
Cell Biology and Genetic Department (University of La Laguna)
Klassert (Helmholtz Centre for Infection Research)
provided critical insights into this study
Borges and the staff of Agrobiology Area of the IPNA-CSIC
Martín (IUETSPC) for lending us some of the equipment used in this study
We sincerely appreciate the contributions of two anonymous reviewers and Dr
which led to a significant improvement of our original manuscript
Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature
This work has been supported by the agreement between the Government of the Canary Islands and CSIC to carry out the project “Habitat use and impacts of the California kingsnake upon native communities of Gran Canaria (Lamproimpact)”
Borja Maestresalas & Marta López-Darias
Marta López-Darias carried out the study conception and design and Mercedes López-González substantially contributed to these tasks
All authors participated in material preparation and data collection
while Mercedes López-González performed genetic analysis
Mercedes López-González and Marta López-Darias wrote the first draft of the manuscript
All authors commented on previous versions of the manuscript and read and approved the final manuscript
Marta López-Darias was in charge of research supervision
project administration and funding acquisition
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DOI: https://doi.org/10.1038/s41598-025-96387-8
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Novel approach to study fish DNA collected from water samples enables researchers to estimate biomass of multiple Alaska fish species simultaneously
Around the world, standard ways to measure fish abundance and biomass to manage commercial, recreational and subsistence fisheries rely heavily on deriving estimates from fish caught in research surveys and commercial fishing nets and other collected biological data. In a new study, NOAA Fisheries scientists
in partnership with the University of Alaska Fairbanks
show that it is possible to estimate fish biomass for more than one species at the same time
They found they can estimate the abundance of ecologically and commercially-important Alaska fish species—Arctic cod
and Pacific cod—which are difficult to distinguish among in eDNA samples
eDNA is genetic material shed by organisms into the surrounding environment
its DNA is shed and accumulates in the water around it
This genetic material can be recovered from environmental samples.
we learned that we could accurately quantify species compositions and estimate biomass for different species of cod and pollock at the same time using eDNA,” said Kimberly Ledger
lead author and research biologist with Alaska Fisheries Science Center’s Auke Bay Laboratories
“It is possible to apply these methods to other species to improve the quantitative utility of eDNA.”
A central goal of managing species is understanding where they live (their distribution) and how many of them exist (their biomass or abundance)
Accurately estimating this information is difficult
It relies on making inferences about an entire community based on observations of a subset of individuals
NOAA Fisheries gets this information from fish collected in nets during research surveys and from data collected by fishery observers on commercial fishing boats and in processing plants.
With eDNA scientists are able to detect and quantify fish DNA from the environment to help estimate species abundance and biomass from just a sample of water
This revolutionary new method that relies on eDNA is highly versatile
capable of detecting and identifying a wide range of species
and minimally disruptive to sensitive species.
Scientists caution that it doesn’t replace standard trawl surveys or actual sampling by fisheries observers
We we still need to collect biological data including size
which is important for stock assessments.
eDNA researchers are actively working to identify ways to use this information to support management decisions with the help of stock assessment authors and quantitative ecologists
A genetic marker is a gene or a short segment of DNA that has a distinctive location on a specific chromosome that can be used to identify a species or an organism.
scientists identified a new genetic marker
to detect and differentiate eDNA from six closely-related cod species in the North Pacific and the Arctic.
Scientists demonstrated that they could
This genetic research also shows promise for estimating the absolute abundance of these fish from eDNA samples.
Fishery biologist and co-author Mary Beth Rew Hicks kept live Pacific cod
and Arctic cod in various combinations and abundances in holding tanks at the Center’s Newport Laboratory
She collected water samples from the tanks with known biomass (based on the numbers and weights of each species of fish)
This experimental design enabled comparison of eDNA-derived species compositions with true compositions within a group of closely-related species with overlapping habitats and distributions.
“We’ve moved the dial in the use of eDNA beyond just being able to provide a measure of presence and absence of species to providing meaningful data to estimate abundance and better reflect underlying community composition in real-world environments,” said Ledger.
fish are moving around to find ideal conditions for their survival—food and the right temperatures to spawn
These dynamic shifts are posing new challenges for resource management and the scientists who conduct long-term research surveys in traditional areas where these fish have been found.
“This new tool will help us efficiently track these shifts in distribution to complement our standard methods of assessing fish stocks,” added Wes Larson
co-author and manager for the Genetics Program at the Alaska Fisheries Science Center.
The family of Edna Marie Johnson created this Life Tributes page to make it easy to share your memories
Receive emails when new obituariesare published to our website
Owned and operated by the Atchley family since 1920
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2025 at Methodist Medical Center in Oak Ridge
She enjoyed being with her family and friends
Edna was sweet and loving to everyone
Jr and Todd Shipley; 12 grandkids and 7 great grandkids
The family will receive friends from 11:00am - 1:00pm
2025 at Mynatt Funeral Home Powell Chapel and then proceed to Norris Memorial Gardens for a 2:00pm graveside service
Mynatt Funeral Home Powell Chapel is honored to be serving the family of Edna Ann Varner
Mynatt Funeral Home Powell Chapel * 2000 Powell Drive * Powell
TN 37849 * (865) 362-5382 * www.mynattfh.com
Edna Katheryn Duncan Cupples passed peacefully on December 29
at the age of 101 in the comforting grace of her family’s love
Katheryn was a force to behold when it came to her faith
She endured over a century of our collective history
She was a devote Catholic and a loving and loyal wife
playful and adventurous in life and in spirit
The pain of her absence is felt by generations
Beloved wife of 64 years to the late Macile Marion Cupples
mother of Marion Annette Cupples Ross and the late Charles Wiley Cupples
Katherine Webb and Benjamin (and Katie) Ross
also members of Immaculate Conception Catholic Church
New Orleans and Brown & Root are invited to attend the Funeral Mass in the chapel of Mothe Funeral Home
Visitation will be held on Thursday from 12pm until 2pm
Interment at Westlawn Memorial Park Cemetery
Family and friends may view and sign the online guest book at www.mothefunerals.com
Edna Adams – Public Relations Specialist in the Office of Communications – was named a Rising Star in PR Daily’s Top Women in Communications awards program
“I’m so honored to receive this recognition for the work I’ve done
but I’m even more honored to do the work for Macon-Bibb County,” says Adams
“The people I work with really make this all so fun and rewarding.”
“Edna has been at the leading edge of changing local government communications
From her writing to her event planning to her media strategy
“What sets Edna apart from her colleagues is her ability to balance all that is required of her…while pushing the envelope on creativity in communications so that the event
the message all stand out in the crowded onslaught of information are bombarded with every day,” says Chief Communications Officer Chris Floore
from original features to public notices to community partner information
from press conferences to groundbreakings to celebrations to major announcements
There are some weeks where she will coordinate 3-4 major events that require significant planning
her events include setting buildings on fire to help the Fire Department make announcements; opening the new pickleball facility on January 1 with a pickleball drop countdown; volunteers packing boxes of food at the Food Bank Grand Opening; reopening an airport runway with planes flying overhead; and more
putting a touch to them to make them more interesting for the media and the public
She has been a key organizer for the community’s Hispanic Heritage Festival
growing it from a few vendors in a small park to having several hundred people
and musical performances in one of our largest parks
“The results speak for themselves,” adds Mayor Miller
showing they saw the coverage and remembered it
The creativity in our event planning means we are getting through the noise and reaching our audience.”
The Office of Communications includes Edna Adams
Their charge is to find the best ways to let people know what the government and its partners are doing to improve the community through strategic communications
These recognitions follow nearly 30 individual
the City-County Communications & Marketing Association (3CMA)
National Association of Government Communicators (NAGC)
and the Georgia chapter of the Solid Waste Association of North America
Customer Service
Sign up for email updates from Macon-Bibb County to get information on announcements
Strategerist host Andrew Kaufmann and Devon Yarbrough
sat down with Milauna Jackson who portrays Captain Abby Campbell in the 2024 Netflix film
“The Six Triple Eight” and Retired Army Col
a passionate advocate whose work led to two crucial recognitions for the 6888
Milauna discusses what it was like working on the Netflix film while Col
Cummings shares the behind-the-scenes bipartisan advocacy for the battalion who played a critical role in World War II
Edna was known for her stubborn yet sweet disposition
embodying a spirit of faithfulness that resonated with all who knew her
Edna found joy in being a dedicated member of Cornerstone Baptist Temple in Dayton
where she cultivated lasting friendships and engaged in the community
She had a passion for bowling and enjoyed playing cards and games
where her competitive yet fun-loving nature shone through
Edna's proudest accomplishments were rooted in her roles as a faithful wife and devoted mother
She taught her daughters the meaning of being a Proverbs 31 wife
Her 68-year marriage to her beloved husband
was a testament to their enduring love and partnership
they raised four children: Richard (Laura) Hamlett
Her love extended to her eight grandchildren and five great-grandchildren
Robert (Rosemary) Gilbert; and numerous nieces and nephews who will fondly remember her as a steadfast presence in their lives
Each of these individuals held a special place in her heart
and their memories will live on through those she leaves behind
but her spirit and the love she shared will endure in the hearts of all who had the privilege of knowing her
The family will receive friends on Thursday
2025 from 1-3pm at the Cornerstone Baptist Temple
with Pastor Jerry Siler officiating.
Edna will be laid to rest at Dayton National Cemetery on Wednesday
It is with great sadness and broken hearts that we announce Edna Mae Wyatt Millisock
“In the blink of an eye she went home to be with her Lord and Savior in her eternal heavenly home.”
Edna was born in the fall season of October 21
which was located in Waynesville at that time
Edna had made her home in Canton all of her life and she was so proud of her family heritage and of her hometown
being involved in things that were happening in Canton
and anything fun that was happening around her
She treasured sharing special memories of her years growing up with her children and these will be memories of hers that they each will hold onto forever
Edna was a beacon of love for her family and for anyone blessed to know her
she chose to enter the health care profession
she was employed as a Certified Nursing Assistant for many dedicated years at Health Care facilities in our community
Edna was like a ray of sunshine to the patients she cared for
and always laughter to brighten their days
Her life has been a testament to her unwavering faith and devotion to her Lord and Saviour
Edna was a faithful member of Old Time Ministries
and with her boundless love for her family
she became an inspiration to each of them to dedicate their lives to Jesus Christ
and we know today that she is surrounded in her heavenly home by the most breathtaking flowers one could ever imagine
She also enjoyed going shopping and “ treasure seeking,” and in her quiet times at home
she could often be found watching her favorite wrestling programs on television
Hearing her lift her sweet voice in praise will never be forgotten by her family
and great-grandmother whose life has touched so very many
let us never forget that a mother’s love lingers like a quiet whisper in our hearts
Edna was preceded in death by the love of her life
Milford Millisock who passed away in 2005; also her sons
Barron Wyatt and Marty White and a granddaughter
Jack (Billy) White and Robert Wyatt; her cherished granddaughter
Brett and grandson Aidan who resided at the home with Edna and where her beloved Laura has devoted her life during Edna’s illness to care for her with great compassion and love; also 18 very loved grandchildren and 12 great-grandchildren who were also adored and cherished by Edna; a sister
A Service of Remembrance to celebrate the life of Edna will be held at 4:00 pm Tuesday
2025 in the chapel of Crawford / Ray Funeral Home and Cremation Service
The family will be receiving friends at the funeral home from 3:00 pm until 4:00 pm prior to the service
Inurnment will be held at a later date at Clarks Chapel Cemetery
where Edna will be laid to rest beside her loved ones
Crawford / Ray Funeral Home and Cremation Service is deeply honored to be caring for the Millisock family
Edna Louise Dowdy, 82, formerly of Chattanooga, passed away Wednesday, March 26, 2025, at her residence in Murfreesboro, Tennessee after a battle with many health complications. A native of Chattanooga, Tennessee, and a resident of LaFayette,... View Obituary & Service Information
The family of Edna Louise Dowdy created this Life Tributes page to make it easy to share your memories
Copyright © 2025 Taylor Funeral Home of Chattanooga
a daughter of the late Henry and Geneva Kirby Sherbert
Edna was married to the late Carl Eggleton.
Edna was a member of First Baptist Church-Spartanburg and was retired from the public school systems.
Mike Wood (Kathy) and Steve Wood (Libby); 6 grandchildren; and 11 great-grandchildren
A cryptside service will be held 11:00 a.m
at Greenlawn Memorial Gardens Heritage Chapel Mausoleum
Greenlawn Memorial Gardens Heritage Chapel Mausoleum
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activities on preceding journals at which the article was previously under consideration are not shown (for instance submission
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