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1.
PLoS One ; 19(2): e0288948, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38359003

RESUMO

Swimmer's itch (SI) is a dermatitis in humans caused by cercariae of avian and mammalian schistosomes which emerge from infected snails on a daily basis. Mitigation methods for SI have long been sought with little success. Copper sulfate application to the water to kill the snail hosts is the historically employed method, but is localized, temporary, and harmful to many aquatic species. Here, we test an alternative method to control Trichobilharzia stagnicolae, a species well-known to cause SI in northern Michigan and elsewhere in North America. Summer relocation of broods of the only known vertebrate host, common merganser (Mergus merganser), greatly reduced snail infection prevalence the following year on two large, geographically separated lakes in northern Michigan. Subsequent years of host relocation achieved and maintained snail infection prevalence at ~0.05%, more than an order of magnitude lower than pre-intervention. A Before-After-Control-Intervention (BACI) study design using multiple-year snail infection data from two intervention lakes and three control lakes demonstrates that dramatic lake-wide reduction of an avian schistosome can be achieved and is not due to natural fluctuations in the parasite populations. The relevance of reducing snail infection prevalence is demonstrated by a large seven-year data set of SI incidence in swimmers at a high-use beach, which showed a substantial reduction in SI cases in two successive years after relocation began. In addition, data from another Michigan lake where vertebrate-host based intervention occurred in the 1980's are analyzed statistically and show a remarkably similar pattern of reduction in snail infection prevalence. Together, these results demonstrate a highly effective SI mitigation strategy that avoids the use of environmentally suspect chemicals and removes incentive for lethal host removal. Biologically, the results strongly suggest that T. stagnicolae is reliant on the yearly hatch of ducklings to maintain populations at high levels on a lake and that the role of migratory hosts in the spring and fall is much less significant.


Assuntos
Dermatite , Schistosomatidae , Esquistossomose , Dermatopatias Parasitárias , Infecções por Trematódeos , Animais , Humanos , Lagos/parasitologia , Infecções por Trematódeos/parasitologia , Esquistossomose/epidemiologia , Dermatopatias Parasitárias/etiologia , Dermatopatias Parasitárias/parasitologia , Patos , Caramujos/parasitologia , Mamíferos
2.
Microbiol Resour Announc ; 13(2): e0108823, 2024 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-38179906

RESUMO

We report genomes of nine phages isolated from Actinobacteria Rhodococcus equi NRRL B-16538. Six of these phages belong to actinobacteriophage cluster CR, which otherwise contains Gordonia phages; two form the CF cluster; and one is a singleton. Genome lengths are 62,017-80,980 bp with 63.9%-67.3% GC content.

3.
Pathogens ; 11(6)2022 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-35745505

RESUMO

A novel schistosome from Planorbella snails currently known as avian schistosomatid sp. C (ASC) was recently described as being capable of causing the papules associated with swimmer's itch. We conducted a paired study with 24 human volunteers, exposing each of their forearms to five drops of water containing cercariae of ASC or Trichobilharzia stagnicolae, and examined the skin for papules 1-3 days later. A mixed effects model showed that only the parasite species significantly affected the number of papules, while prior experimental exposure, swimming history, and swimmer's itch experience did not. The total number of papules produced by the two species were very different: ASC produced a total of 2 papules from the 298 cercariae used, compared to 49 papules from 160 T. stagnicolae cercariae, a difference factor of more than 43X, which was comparable to the odds ratio of 45.5 computed using the statistical model. A well-known agent of swimmer's itch, T. stagnicolae, is able to penetrate human skin more frequently than ASC, likely meaning that ASC is only a minor cause of swimmer's itch where T. stagnicolae is present. We also completed limited experiments that compared the cercarial behavior of the two species in vitro and in situ. A known stimulant of schistosome cercarial penetration, α-linolenic acid, did not stimulate ASC cercariae to initiate penetration-associated behaviors as frequently as T. stagnicolae. However, when placed on esophageal tissue of the known vertebrate host for ASC, Canada goose (Branta canadensis), ASC cercariae were observed penetrating the esophageal epithelium quickly, whereas T. stagnicolae cercariae did not exhibit any penetration behaviors.

4.
Pathogens ; 11(5)2022 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-35631108

RESUMO

Although most studies of digenetic trematodes of the family Schistosomatidae dwell on representatives causing human schistosomiasis, the majority of the 130 identified species of schistosomes infect birds or non-human mammals. The cercariae of many of these species can cause swimmer's itch when they penetrate human skin. Recent years have witnessed a dramatic increase in our understanding of schistosome diversity, now encompassing 17 genera with eight more lineages awaiting description. Collectively, schistosomes exploit 16 families of caenogastropod or heterobranch gastropod intermediate hosts. Basal lineages today are found in marine gastropods and birds, but subsequent diversification has largely taken place in freshwater, with some reversions to marine habitats. It seems increasingly likely that schistosomes have on two separate occasions colonized mammals. Swimmer's itch is a complex zoonotic disease manifested through several different routes of transmission involving a diversity of different host species. Swimmer's itch also exemplifies the value of adopting the One Health perspective in understanding disease transmission and abundance because the schistosomes involved have complex life cycles that interface with numerous species and abiotic components of their aquatic environments. Given the progress made in revealing their diversity and biology, and the wealth of questions posed by itch-causing schistosomes, they provide excellent models for implementation of long-term interdisciplinary studies focused on issues pertinent to disease ecology, the One Health paradigm, and the impacts of climate change, biological invasions and other environmental perturbations.

5.
One Health ; 13: 100280, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34258371

RESUMO

Parasites with complex life cycles engaging multiple host species living among different environments well-exemplify the value of a cross-cutting One Health approach to understanding fundamental concerns like disease emergence or spread. Here we provide new information regarding a pathogenic schistosome trematode parasite of both wild and domestic mammals that has recently expanded its known range from mesic/wet environments of the southeastern United States to the arid southwest. In 2018, 12 dogs living near a man-made pond in Moab, Utah, were found positive for Heterobilharzia americana, the most westerly report of this endemic North American schistosome, and the first from Utah. Raccoon scats collected near the pond were positive for H. americana eggs, and snails living near the pond's water line identified as Galba humilis shed H. americana cercariae, the first indication of natural infections in this widespread North American snail species. The susceptibility of G. humilis to H. americana was confirmed experimentally. Our studies support the existence of two variants of H. americana and emphasize the need for further investigations of lymnaeids and their compatibility with H. americana, to better define the future potential for its spread. Capture of a new species of intermediate host vector snail and construction of man-made habitats suitable for this snail have created the potential for a much more widespread animal health problem, especially for dogs and horses. H. americana will prove difficult to control because of the role of raccoons in maintaining transmission and the amphibious habits of the snail hosts of this pathogenic schistosome.

6.
Int J Parasitol Parasites Wildl ; 7(2): 171-179, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29988865

RESUMO

Histologic studies of fish from Douglas Lake, Cheboygan County, Michigan, USA show that Diplostomum spp. infect the lens of spottail shiners (Notropis hudsonius) and common shiners (Luxilus cornutus). In contrast, infection was confined to the choroidal vasculature of yellow perch (Perca flavescens), and the morphology of the pigment epithelium and retina in regions adjacent to the metacercariae was abnormal. The difference in location of metacercariae within the host suggested that different Diplostomum species may infect shiners and perch in Douglas Lake. Species diversity was investigated by sequencing the barcode region of the cytochrome oxidase I gene of metacercariae. Four species of Diplostomum were identified, all four of which were present in shiner lenses; however, only Diplostomum baeri was present in the perch choroid. To determine whether infection of perch eyes affects the response of the retina to a light stimulus, electroretinograms (ERG) were recorded. The amplitude of the b-wave of the ERG was reduced and the b-wave latency was increased in infected perch, as compared to uninfected eyes, and the flicker-fusion frequency was also reduced. Infection of the yellow perch choroid by Diplostomum baeri, which shows strong host and tissue specificity, has an adverse effect on retinal function, lending support to the hypothesis that parasite-induced impairment of host vision may afford Diplostomum baeri the evolutionary benefit of increasing the likelihood of transmission, via host fish predation, to its definitive avian host.

7.
Bioinformatics ; 32(13): 2026-8, 2016 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-27153674

RESUMO

UNLABELLED: PhamDB is a web application which creates databases of bacteriophage genes, grouped by gene similarity. It is backwards compatible with the existing Phamerator desktop software while providing an improved database creation workflow. Key features include a graphical user interface, validation of uploaded GenBank files, and abilities to import phages from existing databases, modify existing databases and queue multiple jobs. AVAILABILITY AND IMPLEMENTATION: Source code and installation instructions for Linux, Windows and Mac OSX are freely available at https://github.com/jglamine/phage PhamDB is also distributed as a docker image which can be managed via Kitematic. This docker image contains the application and all third party software dependencies as a pre-configured system, and is freely available via the installation instructions provided. CONTACT: snelesen@calvin.edu.


Assuntos
Bacteriófagos/genética , Bases de Dados de Ácidos Nucleicos , Genes Virais , Software , Internet , Linguagens de Programação , Interface Usuário-Computador
8.
CBE Life Sci Educ ; 13(4): 711-23, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25452493

RESUMO

In their 2012 report, the President's Council of Advisors on Science and Technology advocated "replacing standard science laboratory courses with discovery-based research courses"-a challenging proposition that presents practical and pedagogical difficulties. In this paper, we describe our collective experiences working with the Genomics Education Partnership, a nationwide faculty consortium that aims to provide undergraduates with a research experience in genomics through a scheduled course (a classroom-based undergraduate research experience, or CURE). We examine the common barriers encountered in implementing a CURE, program elements of most value to faculty, ways in which a shared core support system can help, and the incentives for and rewards of establishing a CURE on our diverse campuses. While some of the barriers and rewards are specific to a research project utilizing a genomics approach, other lessons learned should be broadly applicable. We find that a central system that supports a shared investigation can mitigate some shortfalls in campus infrastructure (such as time for new curriculum development, availability of IT services) and provides collegial support for change. Our findings should be useful for designing similar supportive programs to facilitate change in the way we teach science for undergraduates.


Assuntos
Genômica/educação , Currículo , Modelos Educacionais , Desenvolvimento de Programas , Estados Unidos , Universidades
9.
CBE Life Sci Educ ; 13(1): 111-30, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24591510

RESUMO

There is widespread agreement that science, technology, engineering, and mathematics programs should provide undergraduates with research experience. Practical issues and limited resources, however, make this a challenge. We have developed a bioinformatics project that provides a course-based research experience for students at a diverse group of schools and offers the opportunity to tailor this experience to local curriculum and institution-specific student needs. We assessed both attitude and knowledge gains, looking for insights into how students respond given this wide range of curricular and institutional variables. While different approaches all appear to result in learning gains, we find that a significant investment of course time is required to enable students to show gains commensurate to a summer research experience. An alumni survey revealed that time spent on a research project is also a significant factor in the value former students assign to the experience one or more years later. We conclude: 1) implementation of a bioinformatics project within the biology curriculum provides a mechanism for successfully engaging large numbers of students in undergraduate research; 2) benefits to students are achievable at a wide variety of academic institutions; and 3) successful implementation of course-based research experiences requires significant investment of instructional time for students to gain full benefit.


Assuntos
Biologia/educação , Currículo , Pesquisa/educação , Atitude , Comportamento Cooperativo , Coleta de Dados , Docentes , Genoma , Genômica/educação , Humanos , Conhecimento , Aprendizagem , Anotação de Sequência Molecular , Avaliação de Programas e Projetos de Saúde , Pesquisadores , Autorrelato , Inquéritos e Questionários , Fatores de Tempo
10.
mBio ; 5(1): e01051-13, 2014 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-24496795

RESUMO

UNLABELLED: Engaging large numbers of undergraduates in authentic scientific discovery is desirable but difficult to achieve. We have developed a general model in which faculty and teaching assistants from diverse academic institutions are trained to teach a research course for first-year undergraduate students focused on bacteriophage discovery and genomics. The course is situated within a broader scientific context aimed at understanding viral diversity, such that faculty and students are collaborators with established researchers in the field. The Howard Hughes Medical Institute (HHMI) Science Education Alliance Phage Hunters Advancing Genomics and Evolutionary Science (SEA-PHAGES) course has been widely implemented and has been taken by over 4,800 students at 73 institutions. We show here that this alliance-sourced model not only substantially advances the field of phage genomics but also stimulates students' interest in science, positively influences academic achievement, and enhances persistence in science, technology, engineering, and mathematics (STEM) disciplines. Broad application of this model by integrating other research areas with large numbers of early-career undergraduate students has the potential to be transformative in science education and research training. IMPORTANCE: Engagement of undergraduate students in scientific research at early stages in their careers presents an opportunity to excite students about science, technology, engineering, and mathematics (STEM) disciplines and promote continued interests in these areas. Many excellent course-based undergraduate research experiences have been developed, but scaling these to a broader impact with larger numbers of students is challenging. The Howard Hughes Medical Institute (HHMI) Science Education Alliance Phage Hunting Advancing Genomics and Evolutionary Science (SEA-PHAGES) program takes advantage of the huge size and diversity of the bacteriophage population to engage students in discovery of new viruses, genome annotation, and comparative genomics, with strong impacts on bacteriophage research, increased persistence in STEM fields, and student self-identification with learning gains, motivation, attitude, and career aspirations.


Assuntos
Bactérias/virologia , Bacteriófagos/genética , Genômica/educação , Microbiologia/educação , Adulto , Feminino , Humanos , Masculino , Estudantes , Adulto Jovem
11.
J Virol ; 88(5): 2461-80, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24335314

RESUMO

UNLABELLED: Genomic analysis of a large set of phages infecting the common host Mycobacterium smegmatis mc(2)155 shows that they span considerable genetic diversity. There are more than 20 distinct types that lack nucleotide similarity with each other, and there is considerable diversity within most of the groups. Three newly isolated temperate mycobacteriophages, Bongo, PegLeg, and Rey, constitute a new group (cluster M), with the closely related phages Bongo and PegLeg forming subcluster M1 and the more distantly related Rey forming subcluster M2. The cluster M mycobacteriophages have siphoviral morphologies with unusually long tails, are homoimmune, and have larger than average genomes (80.2 to 83.7 kbp). They exhibit a variety of features not previously described in other mycobacteriophages, including noncanonical genome architectures and several unusual sets of conserved repeated sequences suggesting novel regulatory systems for both transcription and translation. In addition to containing transfer-messenger RNA and RtcB-like RNA ligase genes, their genomes encode 21 to 24 tRNA genes encompassing complete or nearly complete sets of isotypes. We predict that these tRNAs are used in late lytic growth, likely compensating for the degradation or inadequacy of host tRNAs. They may represent a complete set of tRNAs necessary for late lytic growth, especially when taken together with the apparent lack of codons in the same late genes that correspond to tRNAs that the genomes of the phages do not obviously encode. IMPORTANCE: The bacteriophage population is vast, dynamic, and old and plays a central role in bacterial pathogenicity. We know surprisingly little about the genetic diversity of the phage population, although metagenomic and phage genome sequencing indicates that it is great. Probing the depth of genetic diversity of phages of a common host, Mycobacterium smegmatis, provides a higher resolution of the phage population and how it has evolved. Three new phages constituting a new cluster M further expand the diversity of the mycobacteriophages and introduce novel features. As such, they provide insights into phage genome architecture, virion structure, and gene regulation at the transcriptional and translational levels.


Assuntos
Família Multigênica , Micobacteriófagos/classificação , Micobacteriófagos/genética , Mycobacterium smegmatis/virologia , RNA de Transferência/genética , RNA Viral , Composição de Bases , Sequência de Bases , Códon , Sequência Conservada , Ordem dos Genes , Tamanho do Genoma , Genoma Viral , Sequências Repetidas Invertidas , Lisogenia/genética , Micobacteriófagos/ultraestrutura , Fases de Leitura Aberta , Filogenia , RNA de Transferência/química , Sequências Repetitivas de Ácido Nucleico , Alinhamento de Sequência , Vírion/genética , Vírion/ultraestrutura , Montagem de Vírus/genética
12.
Proc Natl Acad Sci U S A ; 109(28): E1957-62, 2012 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-22623529

RESUMO

Plasmodium falciparum lines differ in their ability to infect mosquitoes. The Anopheles gambiae L3-5 refractory (R) line melanizes most Plasmodium species, including the Brazilian P. falciparum 7G8 line, but it is highly susceptible to some African P. falciparum strains such as 3D7, NF54, and GB4. We investigated whether these lines differ in their ability to evade the mosquito immune system. Silencing key components of the mosquito complement-like system [thioester-containing protein 1 (TEP1), leucine-rich repeat protein 1, and Anopheles Plasmodium-responsive leucine-rich repeat protein 1] prevented melanization of 7G8 parasites, reverting the refractory phenotype. In contrast, it had no effect on the intensity of infection with NF54, suggesting that this line is able to evade TEP1-mediated lysis. When R females were coinfected with a line that is melanized (7G8) and a line that survives (3D7), the coinfection resulted in mixed infections with both live and encapsulated parasites on individual midguts. This finding shows that survival of individual parasites is parasite-specific and not systemic in nature, because parasites can evade TEP1-mediated lysis even when other parasites are melanized in the same midgut. When females from an extensive genetic cross between R and susceptible A. gambiae (G3) mosquitoes were infected with P. berghei, encapsulation was strongly correlated with the TEP1-R1 allele. However, P. falciparum 7G8 parasites were no longer encapsulated by females from this cross, indicating that the TEP1-R1 allele is not sufficient to melanize this line. Evasion of the A. gambiae immune system by P. falciparum may be the result of parasite adaptation to sympatric mosquito vectors and may be an important factor driving malaria transmission.


Assuntos
Anopheles/metabolismo , Anopheles/parasitologia , Proteínas do Sistema Complemento/metabolismo , Proteínas de Insetos/metabolismo , Plasmodium falciparum/metabolismo , Plasmodium falciparum/patogenicidade , Alelos , Animais , Cruzamentos Genéticos , Feminino , Humanos , Sistema Imunitário , Malária/parasitologia , Malária/transmissão , RNA de Cadeia Dupla/genética , Especificidade da Espécie
13.
PLoS One ; 6(10): e26750, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22053209

RESUMO

Five newly isolated mycobacteriophages--Angelica, CrimD, Adephagia, Anaya, and Pixie--have similar genomic architectures to mycobacteriophage TM4, a previously characterized phage that is widely used in mycobacterial genetics. The nucleotide sequence similarities warrant grouping these into Cluster K, with subdivision into three subclusters: K1, K2, and K3. Although the overall genome architectures of these phages are similar, TM4 appears to have lost at least two segments of its genome, a central region containing the integration apparatus, and a segment at the right end. This suggests that TM4 is a recent derivative of a temperate parent, resolving a long-standing conundrum about its biology, in that it was reportedly recovered from a lysogenic strain of Mycobacterium avium, but it is not capable of forming lysogens in any mycobacterial host. Like TM4, all of the Cluster K phages infect both fast- and slow-growing mycobacteria, and all of them--with the exception of TM4--form stable lysogens in both Mycobacterium smegmatis and Mycobacterium tuberculosis; immunity assays show that all five of these phages share the same immune specificity. TM4 infects these lysogens suggesting that it was either derived from a heteroimmune temperate parent or that it has acquired a virulent phenotype. We have also characterized a widely-used conditionally replicating derivative of TM4 and identified mutations conferring the temperature-sensitive phenotype. All of the Cluster K phages contain a series of well conserved 13 bp repeats associated with the translation initiation sites of a subset of the genes; approximately one half of these contain an additional sequence feature composed of imperfectly conserved 17 bp inverted repeats separated by a variable spacer. The K1 phages integrate into the host tmRNA and the Cluster K phages represent potential new tools for the genetics of M. tuberculosis and related species.


Assuntos
Evolução Molecular , Micobacteriófagos/genética , Sítios de Ligação Microbiológicos , Sequência de Bases , Mapeamento Cromossômico , Análise por Conglomerados , Sequência Conservada/genética , Deleção de Genes , Genoma Viral/genética , Anotação de Sequência Molecular , Dados de Sequência Molecular , Família Multigênica/genética , Mutação/genética , Micobacteriófagos/crescimento & desenvolvimento , Micobacteriófagos/isolamento & purificação , Micobacteriófagos/ultraestrutura , Análise de Sequência de DNA , Temperatura , Proteínas Virais/genética , Vírion/genética , Vírion/ultraestrutura , Integração Viral/genética , Replicação Viral/fisiologia
14.
Dev Comp Immunol ; 32(10): 1119-30, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18417215

RESUMO

All fibrinogen (FBG)-bearing proteins documented to date in the freshwater snail Biomphalaria glabrata, the intermediate host of the human blood fluke Schistosoma mansoni, possess the same molecular structure; one or two immunoglobin superfamily (IgSF) domains at the N-terminus and a FBG domain at the C-terminus (named as FBG-related protein (FREP)). Here we report two novel genes that encode FBG-bearing proteins from B. glabrata. Different from all known FREPs, the first gene encodes a protein (657 amino acids (aa)) composed of a long N-terminal region with no sequence homology to any known protein, a middle epidermal growth factor (EGF) repeat region and a C-terminal FBG domain, designated FBG-related molecule (FReM). Differential expression at 2 days post-exposure (dpe) to the trematode S. mansoni or Echinostoma paraensei was found in the S. mansoni susceptible M line and resistant BS-90 snail strains. The second gene is a new member of the FREP family, designated FREP14, which encodes a 399 aa putative secreted protein. FREP14 is different from known FREPs in that it is encoded by a single locus and is not upregulated in early or late stage S. mansoni exposure, but is upregulated in late stage E. paraensei infection. Furthermore, gene expression during the snail's ontogenesis and at a late stage of trematode infection (52 dpe) has been investigated in the two newly identified genes (FReM and FREP14) described in this paper and five representative members of known FREPs (FREPs 2, 3, 4, 12, and 13). A variety of expression patterns were observed, suggestive of functional diversity among the members of FBG-bearing proteins. Our findings further broaden our understanding of the diversity and function of the FBG-bearing protein encoded genes in B. glabrata.


Assuntos
Biomphalaria/genética , Biomphalaria/parasitologia , Echinostoma/fisiologia , Fibrinogênio/metabolismo , Imunoglobulinas/metabolismo , Schistosoma mansoni/fisiologia , Infecções por Trematódeos/transmissão , Sequência de Aminoácidos , Animais , Sequência de Bases , Biomphalaria/química , Biomphalaria/metabolismo , Sequência Conservada , Regulação da Expressão Gênica , Humanos , Imunoglobulinas/química , Imunoglobulinas/genética , Dados de Sequência Molecular , Alinhamento de Sequência , Infecções por Trematódeos/genética , Infecções por Trematódeos/imunologia , Infecções por Trematódeos/parasitologia
15.
J Biol Chem ; 283(6): 3217-3223, 2008 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-18065421

RESUMO

The involvement of reactive oxygen species (ROS) in mosquito immunity against bacteria and Plasmodium was investigated in the malaria vector Anopheles gambiae. Strains of An. gambiae with higher systemic levels of ROS survive a bacterial challenge better, whereas reduction of ROS by dietary administration of antioxidants significantly decreases survival, indicating that ROS are required to mount effective antibacterial responses. Expression of several ROS detoxification enzymes increases in the midgut and fat body after a blood meal. Furthermore, expression of several of these enzymes increases to even higher levels when mosquitoes are fed a Plasmodium berghei-infected meal, indicating that the oxidative stress after a blood meal is exacerbated by Plasmodium infection. Paradoxically, a complete lack of induction of catalase mRNA and lower catalase activity were observed in P. berghei-infected midguts. This suppression of midgut catalase expression is a specific response to ookinete midgut invasion and is expected to lead to higher local levels of hydrogen peroxide. Further reduction of catalase expression by double-stranded RNA-mediated gene silencing promoted parasite clearance by a lytic mechanism and reduced infection significantly. High mosquito mortality is often observed after P. berghei infection. Death appears to result in part from excess production of ROS, as mortality can be decreased by oral administration of uric acid, a strong antioxidant. We conclude that ROS modulate An. gambiae immunity and that the mosquito response to P. berghei involves a local reduction of detoxification of hydrogen peroxide in the midgut that contributes to limit Plasmodium infection through a lytic mechanism.


Assuntos
Anopheles/imunologia , Anopheles/microbiologia , Bactérias/metabolismo , Regulação da Expressão Gênica , Plasmodium berghei/metabolismo , Espécies Reativas de Oxigênio , Animais , Antioxidantes/química , Catalase/metabolismo , Fenômenos Fisiológicos Celulares , Peróxido de Hidrogênio/química , Imunidade Inata , RNA de Cadeia Dupla/química , RNA Mensageiro/metabolismo , Fatores de Tempo
16.
Proc Natl Acad Sci U S A ; 104(7): 2121-6, 2007 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-17284604

RESUMO

The mosquito Anopheles gambiae is a primary vector of Plasmodium parasites in Africa. The effect of aging on reproductive output in A. gambiae females from three strains that differ in their ability to melanize Plasmodium and in their systemic levels of hydrogen peroxide (H2O2), a reactive oxygen species (ROS), was analyzed. The number of eggs oviposited after the first blood meal decreases with age in all strains; however, this decline was much more pronounced in the G3 (unselected) and R (refractory to Plasmodium infection) strains than in the S (highly susceptible to Plasmodium) strain. Reduction of ROS levels in G3 and R females by administration of antioxidants reversed this age-related decline in fecundity. The S and G3 strains were fixed for two functionally different catalase alleles that differ at the second amino acid position (Ser2Trp). Biochemical analysis of recombinant proteins revealed that the Trp isoform has lower specific activity and higher Km than the Ser isoform, indicating that the former is a less efficient enzyme. The Trp-for-Ser substitution appears to destabilize the functional tetrameric form of the enzyme. Both alleles are present in the R strain, and Ser/Ser females had significantly higher fecundity than Trp/Trp females. Finally, a systemic reduction in catalase activity by dsRNA-mediated knockdown significantly reduced the reproductive output of mosquito females, indicating that catalase plays a central role in protecting the oocyte and early embryo from ROS damage.


Assuntos
Anopheles/fisiologia , Catalase/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Envelhecimento , Substituição de Aminoácidos , Animais , Sequência de Bases , Catalase/genética , Catalase/metabolismo , Embrião não Mamífero/enzimologia , Feminino , Fertilidade , Peróxido de Hidrogênio/análise , Dados de Sequência Molecular , Oócitos/enzimologia , Espécies Reativas de Oxigênio/análise
17.
Mem. Inst. Oswaldo Cruz ; 101(supl.1): 167-177, Oct. 2006. tab, graf
Artigo em Inglês | LILACS | ID: lil-441243

RESUMO

To provide a novel resource for analysis of the genome of Biomphalaria glabrata, members of the international Biomphalaria glabrata Genome Initiative (biology.unm.edu/biomphalaria-genome.html), working with the Arizona Genomics Institute (AGI) and supported by the National Human Genome Research Institute (NHGRI), produced a high quality bacterial artificial chromosome (BAC) library. The BB02 strain B. glabrata, a field isolate (Belo Horizonte, Minas Gerais, Brasil) that is susceptible to several strains of Schistosoma mansoni, was selfed for two generations to reduce haplotype diversity in the offspring. High molecular weight DNA was isolated from ovotestes of 40 snails, partially digested with HindIII, and ligated into pAGIBAC1 vector. The resulting B. glabrata BAC library (BG_BBa) consists of 61824 clones (136.3 kb average insert size) and provides 9.05 × coverage of the 931 Mb genome. Probing with single/low copy number genes from B. glabrata and fingerprinting of selected BAC clones indicated that the BAC library sufficiently represents the gene complement. BAC end sequence data (514 reads, 299860 nt) indicated that the genome of B. glabrata contains ~ 63 percent AT, and disclosed several novel genes, transposable elements, and groups of high frequency sequence elements. This BG_BBa BAC library, available from AGI at cost to the research community, gains in relevance because BB02 strain B. glabrata is targeted whole genome sequencing by NHGRI.


Assuntos
Animais , Biomphalaria/genética , Cromossomos Artificiais Bacterianos , Biblioteca Gênica , Schistosoma mansoni/fisiologia , Biomphalaria/classificação , Biomphalaria/parasitologia , Impressões Digitais de DNA , Interações Hospedeiro-Parasita/genética
18.
Mem Inst Oswaldo Cruz ; 101 Suppl 1: 167-77, 2006 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17308766

RESUMO

To provide a novel resource for analysis of the genome of Biomphalaria glabrata, members of the international Biomphalaria glabrata Genome Initiative (http://biology.unm.edu/biomphalaria-genome.html), working with the Arizona Genomics Institute (AGI) and supported by the National Human Genome Research Institute (NHGRI), produced a high quality bacterial artificial chromosome (BAC) library. The BB02 strain B. glabrata, a field isolate (Belo Horizonte, Minas Gerais, Brasil) that is susceptible to several strains of Schistosoma mansoni, was selfed for two generations to reduce haplotype diversity in the offspring. High molecular weight DNA was isolated from ovotestes of 40 snails, partially digested with HindIII, and ligated into pAGIBAC1 vector. The resulting B. glabrata BAC library (BG_BBa) consists of 61824 clones (136.3 kb average insert size) and provides 9.05 x coverage of the 931 Mb genome. Probing with single/low copy number genes from B. glabrata and fingerprinting of selected BAC clones indicated that the BAC library sufficiently represents the gene complement. BAC end sequence data (514 reads, 299860 nt) indicated that the genome of B. glabrata contains ~ 63% AT, and disclosed several novel genes, transposable elements, and groups of high frequency sequence elements. This BG_BBa BAC library, available from AGI at cost to the research community, gains in relevance because BB02 strain B. glabrata is targeted whole genome sequencing by NHGRI.


Assuntos
Biomphalaria/genética , Cromossomos Artificiais Bacterianos , Biblioteca Gênica , Schistosoma mansoni/fisiologia , Animais , Biomphalaria/classificação , Biomphalaria/parasitologia , Impressões Digitais de DNA , Interações Hospedeiro-Parasita/genética
19.
Mol Ecol ; 14(12): 3889-902, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16202103

RESUMO

Schistosoma mansoni is the most widespread of the human-infecting schistosomes, present in 54 countries, predominantly in Africa, but also in Madagascar, the Arabian Peninsula, and the Neotropics. Adult-stage parasites that infect humans are also occasionally recovered from baboons, rodents, and other mammals. Larval stages of the parasite are dependent upon certain species of freshwater snails in the genus Biomphalaria, which largely determine the parasite's geographical range. How S. mansoni genetic diversity is distributed geographically and among isolates using different hosts has never been examined with DNA sequence data. Here we describe the global phylogeography of S. mansoni using more than 2500 bp of mitochondrial DNA (mtDNA) from 143 parasites collected in 53 geographically widespread localities. Considerable within-species mtDNA diversity was found, with 85 unique haplotypes grouping into five distinct lineages. Geographical separation, and not host use, appears to be the most important factor in the diversification of the parasite. East African specimens showed a remarkable amount of variation, comprising three clades and basal members of a fourth, strongly suggesting an East African origin for the parasite 0.30-0.43 million years ago, a time frame that follows the arrival of its snail host. Less but still substantial variation was found in the rest of Africa. A recent colonization of the New World is supported by finding only seven closely related New World haplotypes which have West African affinities. All Brazilian isolates have nearly identical mtDNA haplotypes, suggesting a founder effect from the establishment and spread of the parasite in this large country.


Assuntos
Variação Genética , Filogenia , Schistosoma mansoni/genética , África , Animais , Arábia , Região do Caribe , DNA de Helmintos/genética , DNA Mitocondrial/genética , Feminino , Geografia , Haplótipos , Humanos , Madagáscar , Masculino , Análise de Sequência de DNA , América do Sul
20.
Am J Trop Med Hyg ; 73(1): 131-9, 2005 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16014848

RESUMO

Two species of Biomphalaria are reported from Egypt, the indigenous Biomphalaria alexandrina and Biomphalaria glabrata, the latter believed to be introduced during the past few decades. Both are known to be excellent hosts of Schistosoma mansoni, the human-infecting blood fluke common in Egypt. Given the concerns regarding the spread of the exotic B. glabrata, this study was carried out to get a more current picture of the status of Biomphalaria in Egypt. Snail collections were undertaken during 2002-2003 from regions between Alexandria and Ismailia in the north of the Nile Delta, to as far south as Abu Simbel at Lake Nasser. Biomphalaria snails were found in 37 out of 76 sampled localities and were widely distributed in the Nile Delta and along the Nile as far south as Aswan. According to the results of species-specific polymerase chain reaction assays that sampled both nuclear and mitochondrial genomes, and according to DNA sequence data, all Biomphalaria collected during this survey were B. alexandrina. There was no evidence of the presence of B. glabrata or of hybridization of B. alexandrina with B. glabrata in the examined sites. The results were surprising given that some field-collected snails strongly resembled B. glabrata in both size and conchology and that previous survey work suggested B. glabrata had established in Egypt. Continued scrutiny to ascertain the possible presence of B. glabrata in Egypt is warranted. Also, the planorbid Helisoma duryi was detected in the Delta and as far south as Aswan, so it is important for Egyptian schistosomiasis workers to accurately distinguish this non-schistosome-transmitting snail from Biomphalaria.


Assuntos
Biomphalaria , Animais , Biomphalaria/anatomia & histologia , Biomphalaria/parasitologia , Vetores de Doenças , Egito , Geografia , Densidade Demográfica , Schistosoma mansoni , Esquistossomose mansoni/transmissão
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