Pub Date : 2019-01-01Epub Date: 2019-03-08DOI: 10.1017/pao.2019.1
Mary J Maclean, W Walter Lorenz, Michael T Dzimianski, Christopher Anna, Andrew R Moorhead, Barbara J Reaves, Adrian J Wolstenholme
Lymphatic filariasis (LF) threatens nearly 20% of the world's population and has handicapped one-third of the 120 million people currently infected. Current control and elimination programs for LF rely on mass drug administration of albendazole plus diethylcarbamazine (DEC) or ivermectin. Only the mechanism of action of albendazole is well understood. To gain a better insight into antifilarial drug action in vivo, we treated gerbils harbouring patent Brugia malayi infections with 6 mg kg-1 DEC, 0.15 mg kg-1 ivermectin or 1 mg kg-1 albendazole. Treatments had no effect on the numbers of worms present in the peritoneal cavity of treated animals, so effects on gene expression were a direct result of the drug and not complicated by dying parasites. Adults and microfilariae were collected 1 and 7 days post-treatment and RNA isolated for transcriptomic analysis. The experiment was repeated three times. Ivermectin treatment produced the most differentially expressed genes (DEGs), 113. DEC treatment yielded 61 DEGs. Albendazole treatment resulted in little change in gene expression, with only 6 genes affected. In total, nearly 200 DEGs were identified with little overlap between treatment groups, suggesting that these drugs may interfere in different ways with processes important for parasite survival, development, and reproduction.
{"title":"Effects of diethylcarbamazine and ivermectin treatment on <i>Brugia malayi</i> gene expression in infected gerbils (<i>Meriones unguiculatus</i>).","authors":"Mary J Maclean, W Walter Lorenz, Michael T Dzimianski, Christopher Anna, Andrew R Moorhead, Barbara J Reaves, Adrian J Wolstenholme","doi":"10.1017/pao.2019.1","DOIUrl":"https://doi.org/10.1017/pao.2019.1","url":null,"abstract":"<p><p>Lymphatic filariasis (LF) threatens nearly 20% of the world's population and has handicapped one-third of the 120 million people currently infected. Current control and elimination programs for LF rely on mass drug administration of albendazole plus diethylcarbamazine (DEC) or ivermectin. Only the mechanism of action of albendazole is well understood. To gain a better insight into antifilarial drug action <i>in vivo</i>, we treated gerbils harbouring patent <i>Brugia malayi</i> infections with 6 mg kg<sup>-1</sup> DEC, 0.15 mg kg<sup>-1</sup> ivermectin or 1 mg kg<sup>-1</sup> albendazole. Treatments had no effect on the numbers of worms present in the peritoneal cavity of treated animals, so effects on gene expression were a direct result of the drug and not complicated by dying parasites. Adults and microfilariae were collected 1 and 7 days post-treatment and RNA isolated for transcriptomic analysis. The experiment was repeated three times. Ivermectin treatment produced the most differentially expressed genes (DEGs), 113. DEC treatment yielded 61 DEGs. Albendazole treatment resulted in little change in gene expression, with only 6 genes affected. In total, nearly 200 DEGs were identified with little overlap between treatment groups, suggesting that these drugs may interfere in different ways with processes important for parasite survival, development, and reproduction.</p>","PeriodicalId":87334,"journal":{"name":"Parasitology open","volume":"5 ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1017/pao.2019.1","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25527456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
D. Outlaw, V. W. Walstrom, Haley N. Bodden, Chuan-yu Hsu, Mark A. Arick, D. Peterson
All organisms encounter pathogens, and birds are especially susceptible to infection by malaria parasites and other haemosporidians. It is important to understand how immune genes, primarily innate immune genes which are the first line of host defense, have evolved across birds, a highly diverse group of tetrapods. Here, we find that innate immune genes are highly conserved across the avian tree of life and that although most show evidence of positive or diversifying selection within specific lineages or clades, the number of sites is often proportionally low in this broader context of putative constraint. Rather, the evidence shows a much higher level of negative or purifying selection in these innate immune genes - rather than adaptive immune genes - which is consistent with birds' long coevolutionary history with pathogens and the need to maintain a rapid response to infection. We further explored avian responses to haemosporidians by comparing differential gene expression in wild birds (1) uninfected with haemosporidians, (2) infected with Plasmodium, and (3) infected with Haemoproteus (Parahaemoproteus). We found patterns of significant differential expression with some genes unique to infection with each genus and a few shared between "treatment" groups, but none that overlapped with the genes included in the phylogenetic study.
{"title":"Molecular evolution in immune genes across the avian tree of life.","authors":"D. Outlaw, V. W. Walstrom, Haley N. Bodden, Chuan-yu Hsu, Mark A. Arick, D. Peterson","doi":"10.1017/PAO.2019.3","DOIUrl":"https://doi.org/10.1017/PAO.2019.3","url":null,"abstract":"All organisms encounter pathogens, and birds are especially susceptible to infection by malaria parasites and other haemosporidians. It is important to understand how immune genes, primarily innate immune genes which are the first line of host defense, have evolved across birds, a highly diverse group of tetrapods. Here, we find that innate immune genes are highly conserved across the avian tree of life and that although most show evidence of positive or diversifying selection within specific lineages or clades, the number of sites is often proportionally low in this broader context of putative constraint. Rather, the evidence shows a much higher level of negative or purifying selection in these innate immune genes - rather than adaptive immune genes - which is consistent with birds' long coevolutionary history with pathogens and the need to maintain a rapid response to infection. We further explored avian responses to haemosporidians by comparing differential gene expression in wild birds (1) uninfected with haemosporidians, (2) infected with Plasmodium, and (3) infected with Haemoproteus (Parahaemoproteus). We found patterns of significant differential expression with some genes unique to infection with each genus and a few shared between \"treatment\" groups, but none that overlapped with the genes included in the phylogenetic study.","PeriodicalId":87334,"journal":{"name":"Parasitology open","volume":"5 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1017/PAO.2019.3","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"57048610","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-01-01Epub Date: 2018-11-06DOI: 10.1017/pao.2018.15
Yousuf A Khan, Norma W Andrews, Bidyottam Mittra
Leishmania virulence and disease development critically depends on the ability of Leishmania promastigotes to infect, differentiate into amastigote forms and replicate inside mammalian host macrophages. Understanding changes associated with amastigote differentiation in axenic culture conditions is key to identifying virulence factors. Here we compared efficiency of the conventional pH-temperature-dependent shift method to induce amastigote differentiation with the recently identified trigger for differentiation mediated by mitochondrial reactive oxygen species (ROS). Using two different visceral leishmaniasis species, L. infantum and. L. donovani, we show that ROS-generating methods such as iron deprivation or exposure to sub-lethal concentrations of H2O2 or menadione are significantly more effective in promoting promastigoteamastigote differentiation than the low pH-high temperature shift, leading to higher survival rates, morphological changes and gene expression patterns characteristic of the amastigote stage. Notably, both H2O2 and menadione-mediated differentiation did not require up-regulation of the mitochondrial electron transport chain (ETC)-associated protein p27, suggesting that treatment with oxidants bypasses the necessity to upregulate mitochondrial activity, a precondition for mROS generation. Our findings confirm that ROS-induced differentiation occurs in multiple Leishmania species, including the medically important visceralizing species, and provide mechanistic rationale for earlier reports demonstrating markedly increased virulence of L. infantum promastigotes pre-treated with oxidative reagents.
{"title":"ROS regulate differentiation of visceralizing <i>Leishmania</i> species into the virulent amastigote form.","authors":"Yousuf A Khan, Norma W Andrews, Bidyottam Mittra","doi":"10.1017/pao.2018.15","DOIUrl":"https://doi.org/10.1017/pao.2018.15","url":null,"abstract":"<p><p><i>Leishmania</i> virulence and disease development critically depends on the ability of <i>Leishmania</i> promastigotes to infect, differentiate into amastigote forms and replicate inside mammalian host macrophages. Understanding changes associated with amastigote differentiation in axenic culture conditions is key to identifying virulence factors. Here we compared efficiency of the conventional pH-temperature-dependent shift method to induce amastigote differentiation with the recently identified trigger for differentiation mediated by mitochondrial reactive oxygen species (ROS). Using two different visceral leishmaniasis species, <i>L. infantum</i> and. <i>L. donovani</i>, we show that ROS-generating methods such as iron deprivation or exposure to sub-lethal concentrations of H<sub>2</sub>O<sub>2</sub> or menadione are significantly more effective in promoting promastigoteamastigote differentiation than the low pH-high temperature shift, leading to higher survival rates, morphological changes and gene expression patterns characteristic of the amastigote stage. Notably, both H<sub>2</sub>O<sub>2</sub> and menadione-mediated differentiation did not require up-regulation of the mitochondrial electron transport chain (ETC)-associated protein p27, suggesting that treatment with oxidants bypasses the necessity to upregulate mitochondrial activity, a precondition for mROS generation. Our findings confirm that ROS-induced differentiation occurs in multiple <i>Leishmania</i> species, including the medically important visceralizing species, and provide mechanistic rationale for earlier reports demonstrating markedly increased virulence of <i>L. infantum</i> promastigotes pre-treated with oxidative reagents.</p>","PeriodicalId":87334,"journal":{"name":"Parasitology open","volume":"4 ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1017/pao.2018.15","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37242482","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-01-01Epub Date: 2018-08-02DOI: 10.1017/pao.2018.12
Helen E Robertson, Philipp H Schiffer, Maximilian J Telford
The Dicyemida and Orthonectida are two groups of tiny, simple, vermiform parasites that have historically been united in a group named the Mesozoa. Both Dicyemida and Orthonectida have just two cell layers and appear to lack any defined tissues. They were initially thought to be evolutionary intermediates between protozoans and metazoans but more recent analyses indicate that they are protostomian metazoans that have undergone secondary simplification from a complex ancestor. Here we describe the first almost complete mitochondrial genome sequence from an orthonectid, Intoshia linei, and describe nine and eight mitochondrial protein-coding genes from Dicyema sp. and Dicyema japonicum, respectively. The 14,247 base pair long I. linei sequence has typical metazoan gene content, but is exceptionally AT-rich, and has a unique gene order. The data we have analysed from the Dicyemida provide very limited support for the suggestion that dicyemid mitochondrial genes are found on discrete mini-circles, as opposed to the large circular mitochondrial genomes that are typical of the Metazoa. The cox1 gene from dicyemid species has a series of conserved, in-frame deletions that is unique to this lineage. Using cox1 genes from across the genus Dicyema, we report the first internal phylogeny of this group.
{"title":"The mitochondrial genomes of the mesozoans <i>Intoshia linei, Dicyema</i> sp. and <i>Dicyema japonicum</i>.","authors":"Helen E Robertson, Philipp H Schiffer, Maximilian J Telford","doi":"10.1017/pao.2018.12","DOIUrl":"https://doi.org/10.1017/pao.2018.12","url":null,"abstract":"<p><p>The Dicyemida and Orthonectida are two groups of tiny, simple, vermiform parasites that have historically been united in a group named the Mesozoa. Both Dicyemida and Orthonectida have just two cell layers and appear to lack any defined tissues. They were initially thought to be evolutionary intermediates between protozoans and metazoans but more recent analyses indicate that they are protostomian metazoans that have undergone secondary simplification from a complex ancestor. Here we describe the first almost complete mitochondrial genome sequence from an orthonectid, <i>Intoshia linei</i>, and describe nine and eight mitochondrial protein-coding genes from <i>Dicyema</i> sp. and <i>Dicyema japonicum</i>, respectively. The 14,247 base pair long <i>I. linei</i> sequence has typical metazoan gene content, but is exceptionally AT-rich, and has a unique gene order. The data we have analysed from the Dicyemida provide very limited support for the suggestion that dicyemid mitochondrial genes are found on discrete mini-circles, as opposed to the large circular mitochondrial genomes that are typical of the Metazoa. The <i>cox1</i> gene from dicyemid species has a series of conserved, in-frame deletions that is unique to this lineage. Using <i>cox1</i> genes from across the genus <i>Dicyema</i>, we report the first internal phylogeny of this group.</p>","PeriodicalId":87334,"journal":{"name":"Parasitology open","volume":"4 ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1017/pao.2018.12","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36393563","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2018-01-01Epub Date: 2018-01-11DOI: 10.1017/pao.2017.17
Carolina R Marotta, Priscilla N Dos Santos, Matheus D Cordeiro, Juliana Helena Da S Barros, Lesley Bell-Sakyi, Adivaldo H Fonseca
Parasites of the genus Trypanosoma are microorganisms that display wide morphological, biological and genetic variability. Here we present the first description of an isolate of the genus Trypanosoma naturally infecting the tick Amblyomma brasiliense. The ticks were collected from a specimen of Tayassu pecari (Queixada, white-lipped peccary) from the Itatiaia National Park, Itatiaia, Rio de Janeiro, Brazil. The isolate was characterised by molecular, morphometric and biological analyses. A Trypanosoma culture was isolated from crushed nymphal and adult ticks, propagated in the tick cell line IDE8 and maintained in L15B culture medium, incubated at 32 °C. The isolate grew well in L15B medium at 30 °C, 32 °C and 34 °C but not at lower or higher temperatures. The culture remained stable in axenic L15B medium at 30 °C. Cryopreserved cultures retained viability after cryopreservation in liquid nitrogen. Growth in axenic medium and developmental forms of the trypanosomes were analysed. Analysis of the 18S rDNA region confirmed the authenticity of this new species and the nucleotide sequence was deposited in Genbank. The species was named Trypanosoma amblyommi sp. nov. strain C1RJ. Characteristics related to pathogenicity, involvement with vertebrate hosts, epidemiology, developmental cycle and transmission mechanisms are still unknown. Therefore, further studies are necessary to understand aspects of the biological cycle of Trypanosoma amblyommi sp. nov.
{"title":"<i>Trypanosoma amblyommi</i> sp. nov. (Protozoa: Kinetoplastida) isolated from <i>Amblyomma brasiliense</i> (Acari: Ixodidae) ticks in Rio de Janeiro, Brazil.","authors":"Carolina R Marotta, Priscilla N Dos Santos, Matheus D Cordeiro, Juliana Helena Da S Barros, Lesley Bell-Sakyi, Adivaldo H Fonseca","doi":"10.1017/pao.2017.17","DOIUrl":"10.1017/pao.2017.17","url":null,"abstract":"<p><p>Parasites of the genus <i>Trypanosoma</i> are microorganisms that display wide morphological, biological and genetic variability. Here we present the first description of an isolate of the genus <i>Trypanosoma</i> naturally infecting the tick <i>Amblyomma brasiliense</i>. The ticks were collected from a specimen of <i>Tayassu pecari</i> (Queixada, white-lipped peccary) from the Itatiaia National Park, Itatiaia, Rio de Janeiro, Brazil. The isolate was characterised by molecular, morphometric and biological analyses. A <i>Trypanosoma</i> culture was isolated from crushed nymphal and adult ticks, propagated in the tick cell line IDE8 and maintained in L15B culture medium, incubated at 32 °C. The isolate grew well in L15B medium at 30 °C, 32 °C and 34 °C but not at lower or higher temperatures. The culture remained stable in axenic L15B medium at 30 °C. Cryopreserved cultures retained viability after cryopreservation in liquid nitrogen. Growth in axenic medium and developmental forms of the trypanosomes were analysed. Analysis of the 18S rDNA region confirmed the authenticity of this new species and the nucleotide sequence was deposited in Genbank. The species was named <i>Trypanosoma amblyommi</i> sp. nov. strain C1RJ. Characteristics related to pathogenicity, involvement with vertebrate hosts, epidemiology, developmental cycle and transmission mechanisms are still unknown. Therefore, further studies are necessary to understand aspects of the biological cycle of <i>Trypanosoma amblyommi</i> sp. nov.</p>","PeriodicalId":87334,"journal":{"name":"Parasitology open","volume":"4 ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5837022/pdf/emss-76271.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35893558","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}