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He maintains a general litigation practice concentrating in mass tort and industry-wide litigation, as well as matters involving class actions, products liability, negligence and insurance. He has represented a wide variety of defendants in asbestos personal injury and property damage litigation in Maryland and nationally since He has tried dozens of civil jury trials to verdict in cases lasting from a week to nearly seven months. He has briefed and argued over 60 reported appellate decisions.
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Try out PMC Labs and tell us what you think. Learn More. Box , Nairobi, Kenya. Echinostomes are a diverse group of digenetic trematodes that are globally distributed.
The diversity of echinostomes in Africa remains largely unknown, particularly in analyses using molecular markers. Therefore, we were interested in the composition and host usage patterns of African echinostomes, especially those that also use schistosome transmitting snails as intermediate hosts.
We collected adults and larval stages of echinostomes from 19 different localities in East Africa 1 locality in Uganda and 18 in Kenya. In this study we provide locality information, host use, museum vouchers, and genetic data for two loci 28S and nad 1 from 98 samples of echinostomes from East Africa.
Combining morphological features, host use information, and phylogenetic analyses we found 17 clades of echinostomes in East Africa. Four clades were found to use more than one genus of freshwater snails as their first intermediate hosts. We also determined at least partial life cycles 2 of the 3 of four clades using molecular markers. Of the 17 clades, 13 use Biomphalaria or Bulinus as a first intermediate host. The overlap in host usage creates opportunities for competition, including against human schistosomes.
Thus, our study can be used as a foundation for future studies to ascertain the interactions between schistosomes and echinostomes in their respective intermediate hosts. The Echinostomatoidea is a diverse superfamily of trematodes that includes nine families and genera Tkach et al.
Here we focus on taxa from one of the families the Echinostomatidae, referred to hereafter as echinostomes. Echinostomes are globally distributed and have a multi-host life cycle that involves a vertebrate definitive host, a molluscan first intermediate host, and a second intermediate host that is typically a mollusc, amphibian, or fish.
Echinostomes are known to cause disease in humans, mostly in southeast Asia where raw second intermediate hosts are consumed Graczyk and Fried, Echinostomes are also known to actively influence the establishment of pre-existing infections in snail first intermediate hosts, thus are considered important components to community composition over time and space Lim and Heyneman, ; Sousa, ; Lafferty et al.
Echinostomes are characterized by having a distinctive cephalic crown of collar spines, a ventral sucker larger than the oral sucker, two testes tandemly or symmetrically arranged, a pretesticular ovary, and a cirrus sac Fried, ; Kostadinova and Jones, ; Fried and Toledo, The family Echinostomatidae the recent reclassification now includes taxa that belonged to the former Rhopaliidae, Looss, ; Cathaemasiidae Fuhrmann, ; and Ribeiroiinae Travassos, is the most speciose family in the superfamily Tkach et al.
Delineation of genera has traditionally been based extensively on characteristics of adult worms and has included consideration of definitive host use, the morphology of the cephalic collar, number and arrangement of the collar spines, position of the testes and ovary, and location and structure of the vitellaria Kostadinova, Characteristics of the larval stages, especially of cercariae, have received less consideration.
A recent molecular phylogenetic study focused on 28S rRNA sequences and incorporated a broad array of echinostome species has provided a new framework to organize our thinking about echinostomes Tkach et al.
The focus of this study is to uncover the diversity of echinostomes transmitted in Africa as part of a larger focus in how biodiversity can influence disease transmission.
One of our motivations is to learn how other digenean species, particularly echinostomes, can influence schistosomiasis transmission in Sub-Saharan Africa by competing with schistosome sporocysts for access to their required snail hosts. Studies have shown that when multiple species of digenetic trematodes colonize the same snail host, echinostomes have usually proven to be dominant to other species Lim and Heyneman, ; Hechinger et al.
There is growing evidence for some species that echinostome rediae are specialized for the purpose of attacking and killing the larvae of competing digenean species within the body of their molluscan hosts, while other rediae are more specialized for reproduction Garcia-Vedrenne et al.
An important first step in understanding echinostome-schistosome interactions is the characterization of the biodiversity and host use of echinostome species in East Africa. With these data, we can relate our results to the growing body of research that highlights the relevance between biodiversity and human disease transmission Johnson and Thieltges, ; Civitello et al. However, very little is known about African echinostomes with respect to biogeography, phylogenetic placement especially using molecular markers , and host use.
The majority of echinostome descriptions from Africa are of adults that use birds as a definitive host Dietz, ; Odhner, ; Faust, ; Hilmy, ; Dollfus, ; Bisseru, ; Appleton et al. Even meticulous species descriptions of adult morphology alone can lead to confusion in their systematics.
Likewise, descriptions other life cycle stages like cercariae or metacercariae by themselves may also prove difficult or unreliable as a basis for species identifications. Therefore, to characterize the species diversity of echinostomes in Kenya and surrounding areas, particularly in the Lake Victoria Basin which encompasses multiple East African countries, molecular markers were used in combination with morphological features to identify the life cycle stages we collected.
This effort allowed us to link certain life cycle stages across space and time to better differentiate clades of echinostomes and understand host usage patterns and how they relate to disease transmission in East Africa. Towards this end, we collected and characterized different species of echinostomes that are transmitted in East Africa, primarily from western Kenya, with an emphasis on species that use Biomphalaria or Bulinus as their first intermediate hosts.
These two snail genera host Schistosoma mansoni, and S. Our goal here is to detail how many echinostome species use these snail hosts to provide context for future experiments to determine their ability to compete with and prey upon the sporocysts of schistosomes in their snail hosts.
It is also critical for the evolutionary characterization of organisms to have a permanent museum voucher Pleijel et al. The tissue culture plates were placed in natural light for two hours to induce shedding of cercariae. Available keys were used for preliminary identification of African snails and their trematodes Fain, ; Brown ; Brown and Kristensen, ; Brown, ; Frandsen and Christensen, ; Schell, To maintain a connection between the morphological voucher and the genetic voucher, part of the posterior portion of the adult was severed and used for molecular work and the remaining part was stained according to Fried and Manger We sequenced the 28S gene because such data are available for many of the echinostomes listed in GenBank and can thus provide a broader taxonomic comparison of our specimens into genera according to the scheme of Tkach et al.
We sequenced the nad 1 gene to provide additional resolution for some of the more-closely related representatives we obtained. Partial sequences of the 28S ribosomal gene and nicotinamide adenine dinucleotide dehydrogenase subunit 1 nad 1 from 98 echinostome specimens were amplified by polymerase chain reaction PCR.
Samples were chosen based on host usage, locality, and sampling year. One or two cercariae, one rediae, or a partial portion of the posterior end of an adult were used for DNA extraction. PCR cycles were followed according to Tkach et al. For some of the samples, only the cercariae after shedding of snails were saved, but its snail host was not preserved. Cercariae can often have adherent snail tissue on them that can be amplified with snail specific primers Devkota et al.
Therefore, for the cercariae where we did not have the snail host, we used snail cox1 primers to generate amplicons from those cercariae, particularly those shed from Bulinus. This was done in attempt to verify the Bulinus species from which the cercariae were shed, because identification based only on snail keys is difficult for this genus Stothard et al. Many of samples yielded amplicons; however, in some cases, we were unable to amplify snail DNA from the cercariae samples, therefore we could not designate a species.
PCR fragments were separated by agarose gel electrophoresis and visualized with 0. Both strands were sequenced using an Applied Biosystems automated sequencer and BigDye terminator cycle sequencing kit Version 3. A total of bases were used for 28S alignment and bases for nad 1 alignments. Sequences generated in this study were submitted to GenBank Table 2.
Heuristic searchers were utilized for ML analyses and bootstrap replicates were run for each dataset. Provisional identification, sample name, host it was collected from, life cycle stage, collection locality, date, Museum of Southwestern Biology voucher number, and GenBank accession numbers of echinostome specimens used in this study.
Data were summarized within and between groups Table 3 , Table 4. Intra- and interclade P- distance values of 28S amplified from the 98 echinostomes in this study. Intra- and interclade P- distance values of nad1 amplified from the 94 minus the 4 Ribeiroia samples echinostomes in this study. We collected echinostome adults and larva between — from 19 localities Table 1.
Cercariae or rediae were collected from 9 species of snail hosts and adults were collected from two species of birds. We sequenced 28S and nad 1 from 92 different cercariae, 4 metacercariae, and 2 adult samples. Although we attempted to sequence nad 1 from all 98 samples, 4 samples would not amplify using the nad 1 primers.
Forty-seven samples from GenBank and 98 specimens from this study were used in phylogenetic analyses to determine how the samples were related. Because some of our resulting clades had multiple representatives, we chose two or three specimens per clade to simplify the presentation of echinostome diversity.
Sequences bp were obtained for all 98 samples of which bp were used for Maximum Likelihood Fig. Caballerotrema sp. Tkach et al. These analyses revealed 17 clades, the names for which are shown in Fig. Clades were color coded Fig. Phylogenetic relationships of echinostomes from this and study bolded and from GenBank with accession numbers based on bp of the 28S gene inferred from ML and BI analyses.
Specimens are named based on sample name, the host and locality it was collected from, and color-coded based on clade designation from nad 1 p-distance values of less than 1. A black circle indicates clades where more than one genus of snails was found to be infected and a red star indicates clades where sequences from two different life-cycle stages matched.
Phylogenetic relationships of echinostomes from this and study bolded and from GenBank with accession numbers based on bp of the nad 1 gene inferred from ML and BI analyses. Pictures of echinostomoid cercariae collected from Kenya: Clade 3, Patagifer sp.
A2 represents the cluster of spines posterior to the oral sucker, B clade 1, Patagifer sp. Forty-one samples from GenBank and the same specimens from this study were used to generate the 28S tree in this study were used in the analysis. Four of the Ribeiroia samples did not amplify or the quality of the sequences was poor. Therefore, 94 samples were used in the original analyses and to determine p-distance values. Nad 1 sequences revealed two additional clades that were not found from the 28S analysis see below under Patagifer.
Species of Patagifer were known to use ibises as definitive hosts and snails as both the first and second intermediate hosts Faltynkova et al. Many of our samples 43 grouped into clades 2 and 3, including 2 samples from Uganda. Thirty-one specimens grouped into clade 2 Patagifer sp. There was a 0. We completed the life cycle of worms from clade 2. We acquired eggs from fecal samples from a sacred ibis Threskiornis aethiopicus , hatched the eggs and experimentally exposed Biomphalaria sudanica to the miracidia.
We then used cercariae from successful experimental infections to expose B. We sequenced representatives of each life cycle stage for clade 2 and found them to be identical or to differ by less than 1. Clade 2 cercariae had tail fins and 58—62 collar spines. The larvae also possessed a structure we termed the spine pocket containing approximately 20 spines that was located mid-ventrally just posterior to the oral sucker.
Acquisition of nad 1 sequences for P. We also note of interest that our cercariae in clade 2 resembled cercariae from two South American species of Biomphalaria : 1 cercariae of B.
Samples from Kazinga Channel in Uganda also grouped into this clade and clade 3. Clade 3 cercariae have tail fins, 54 collar spines, a spine pocket posterior to the oral sucker containing a cluster of 25 spines Fig. Appleton et al.
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A dengue vaccine for individuals aged 9 years and older has been licensed, but there remains urgent medical need for a vaccine that is safe and effective against all four dengue virus serotypes DENV in recipients of all ages. Company officials, however, are confident they'll get it. Takeda's product is the second dengue vaccine to enter clinical trials, after Sanofi Pasteur's Dengvaxia. New long-term Phase III data for Takeda's candidate dengue vaccine show continued protection against illness and hospitalization, regardless of previous exposure, out to the three-year mark following the two-dose jab. Read more.
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State Rep. Aaron Miller took advantage of an opportunity Friday to tour and learn more about the St. Joseph County Health Center. The freshman representative heard details about the Centreville-based agency from Lisa Loker, marketing director, and Steve Kujacznski, medical director of the facility. Following a tour of the building, Miller met with Loker, Kujacznski and board member Gwen Donmyer to hear more about its services and how it is meeting the needs of St. Joseph County residents. Loker said the center is eager to hear more about potential legislation regarding registered nurse advanced practice.
The Covid pandemic has put the UK health and care workforce under unprecedented pressure. The workforce had been struggling to cope even before the pandemic took hold. Staff stress, absenteeism, turnover and intentions to quit had reached alarmingly high levels in , with large numbers of nurse and midwife vacancies across the health and care system.
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