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Article: Theileria

Woods, Kerry / Perry, Carmen / Brühlmann, Francis / Olias, Philipp

Frontiers in cell and developmental biology

2021 Volume 9, Page(s) 662805

Abstract: One of the first events that follows invasion of leukocytes ... ...

Abstract	One of the first events that follows invasion of leukocytes by
Language	English
Publishing date	2021-04-20
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2737824-X
ISSN	2296-634X
ISSN	2296-634X
DOI	10.3389/fcell.2021.662805
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Article ; Online: Functional Analysis of the Role of Toxoplasma gondii Nucleoside Triphosphate Hydrolases I and II in Acute Mouse Virulence and Immune Suppression.

Olias, Philipp / Sibley, L David

Infection and immunity

2016 Volume 84, Issue 7, Page(s) 1994–2001

Abstract: Bioluminescent reporter assays have been widely used to study the effect of Toxoplasma gondii on host gene expression. In the present study, we extend these studies by engineering novel reporter cell lines containing a gamma-activated sequence (GAS) ... ...

Abstract	Bioluminescent reporter assays have been widely used to study the effect of Toxoplasma gondii on host gene expression. In the present study, we extend these studies by engineering novel reporter cell lines containing a gamma-activated sequence (GAS) element driving firefly luciferase (FLUC). In RAW264.7 macrophages, T. gondii type I strain (GT1) infection blocked interferon gamma (IFN-γ)-induced FLUC activity to a significantly greater extent than infection by type II (ME49) and type III (CTG) strains. Quantitative trait locus (QTL) analysis of progeny from a prior genetic cross identified a genomic region on chromosome XII that correlated with the observed strain-dependent phenotype. This QTL region contains two isoforms of the T. gondii enzyme nucleoside triphosphate hydrolase (NTPase) that were the prime candidates for mediating the observed strain-specific effect. Using reverse genetic analysis we show that deletion of NTPase I from a type I strain (RH) background restored the higher luciferase levels seen in the type II (ME49) strain. Rather than an effect on IFN-γ-dependent transcription, our data suggest that NTPase I was responsible for the strain-dependent difference in FLUC activity due to hydrolysis of ATP. We further show that NTPases I and II were not essential for tachyzoite growth in vitro or virulence in mice. Our study reveals that although T. gondii NTPases are not essential for immune evasion, they can affect ATP-dependent reporters. Importantly, this limitation was overcome using an ATP-independent Gaussia luciferase, which provides a more appropriate reporter for use with T. gondii infection studies.
MeSH term(s)	Acute Disease ; Adenosine Triphosphate/metabolism ; Animals ; Cell Line ; Chromosome Mapping ; Gene Expression ; Gene Knockout Techniques ; Genes, Reporter ; Host-Parasite Interactions/genetics ; Host-Parasite Interactions/immunology ; Humans ; Immunomodulation ; Mice ; Mutation ; Nucleoside-Triphosphatase/genetics ; Nucleoside-Triphosphatase/metabolism ; Quantitative Trait Loci ; STAT1 Transcription Factor/genetics ; STAT1 Transcription Factor/metabolism ; Toxoplasma/enzymology ; Toxoplasma/genetics ; Toxoplasma/immunology ; Toxoplasma/pathogenicity ; Toxoplasmosis, Animal/immunology ; Toxoplasmosis, Animal/parasitology ; Transcription, Genetic ; Virulence/genetics
Chemical Substances	STAT1 Transcription Factor ; Adenosine Triphosphate (8L70Q75FXE) ; Nucleoside-Triphosphatase (EC 3.6.1.15)
Language	English
Publishing date	2016-07
Publishing country	United States
Document type	Journal Article
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
DOI	10.1128/IAI.00077-16
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Article: Fatal Diplostomum phoxini infection in captive Atlantic puffin Fratercula arctica chicks following ingestion of infected European minnows Phoxinus phoxinus.

Hoby, Stefan / Berenguer Veiga, Inês / Olias, Philipp / Küchler, Leonore / Schönbächler, Katja / Blatter, Hansueli / Schmidt-Posthaus, Heike

Diseases of aquatic organisms

2022 Volume 150, Page(s) 161–167

Abstract: Conservation of endangered animal species is a major task of zoos. Husbandry and breeding of Atlantic puffins Fratercula arctica in captivity is challenging. In 2019, the entire chick population (n = 4 chicks) in Berne Animal Park's Atlantic puffin ... ...

Abstract	Conservation of endangered animal species is a major task of zoos. Husbandry and breeding of Atlantic puffins Fratercula arctica in captivity is challenging. In 2019, the entire chick population (n = 4 chicks) in Berne Animal Park's Atlantic puffin colony (Bern, Switzerland) died within 7 d. Due to supply constraints, the chicks had been fed with wild-caught European minnows Phoxinus phoxinus. At necropsy, the main pathological finding in all deceased puffin chicks was a multifocal, moderate to severe subacute heterophilic and granulomatous enteritis with intralesional adult trematodes and eggs. Metacercariae surrounded by few necrotic cells and scattered macrophages were found in the brain and spinal cord of the food fish. Additional microbiological analyses of both the puffin chicks and fish were unremarkable. Diplostomum phoxini DNA could be identified in formalin-fixed paraffin-embedded tissue from the small intestine of all puffin chicks and European minnows following PCR and sequencing of the 18S ribosomal RNA gene and the internal transcribed spacer (ITS1) region. This report illustrates the importance of intensive health checks of food fish for animal species kept in captivity.
MeSH term(s)	Animals ; Brain ; Charadriiformes ; Cyprinidae ; Eating ; Trematoda
Language	English
Publishing date	2022-08-18
Publishing country	Germany
Document type	Journal Article
ISSN	0177-5103
ISSN	0177-5103
DOI	10.3354/dao03683
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Article ; Online: Single- and duplex TaqMan-quantitative PCR for determining the copy numbers of integrated selection markers during site-specific mutagenesis in Toxoplasma gondii by CRISPR-Cas9.

Hänggeli, Kai Pascal Alexander / Hemphill, Andrew / Müller, Norbert / Schimanski, Bernd / Olias, Philipp / Müller, Joachim / Boubaker, Ghalia

PloS one

2022 Volume 17, Issue 9, Page(s) e0271011

Abstract: Herein, we developed a single and a duplex TaqMan quantitative PCR (qPCR) for absolute quantification of copy numbers of integrated dihydrofolate reductase-thymidylate synthase (mdhfr-ts) drug selectable marker for pyrimethamine resistance in Toxoplasma ... ...

Abstract	Herein, we developed a single and a duplex TaqMan quantitative PCR (qPCR) for absolute quantification of copy numbers of integrated dihydrofolate reductase-thymidylate synthase (mdhfr-ts) drug selectable marker for pyrimethamine resistance in Toxoplasma gondii knockouts (KOs). The single TaqMan qPCR amplifies a 174 bp DNA fragment of the inserted mdhfr-ts and of the wild-type (WT) dhfr-ts (wtdhfr-ts) which is present as single copy gene in Toxoplasma and encodes a sensitive enzyme to pyrimethamine. Thus, the copy number of the dhfr-ts fragment in a given DNA quantity from KO parasites with a single site-specific integration should be twice the number of dhfr-ts copies recorded in the same DNA quantity from WT parasites. The duplex TaqMan qPCR allows simultaneous amplification of the 174 bp dhfr-ts fragment and the T. gondii 529-bp repeat element. Accordingly, for a WT DNA sample, the determined number of tachyzoites given by dhfr-ts amplification is equal to the number of tachyzoites determined by amplification of the Toxoplasma 529-bp, resulting thus in a ratio of 1. However, for a KO clone having a single site-specific integration of mdhfr-ts, the calculated ratio is 2. We then applied both approaches to test T. gondii RH mutants in which the major surface antigen (SAG1) was disrupted through insertion of mdhfr-ts using CRISPR-Cas9. Results from both assays were in correlation showing a high accuracy in detecting KOs with multiple integrated mdhfr-ts. Southern blot analyses using BsaBI and DraIII confirmed qPCRs results. Both TaqMan qPCRs are needed for reliable diagnostic of T. gondii KOs following CRISPR-Cas9-mediated mutagenesis, particularly with respect to off-target effects resulting from multiple insertions of mdhfr-ts. The principle of the duplex TaqMan qPCR is applicable for other selectable markers in Toxoplasma. TaqMan qPCR tools may contribute to more frequent use of WT Toxoplasma strains during functional genomics.
MeSH term(s)	Antigens, Surface/pharmacology ; CRISPR-Cas Systems/genetics ; DNA/pharmacology ; DNA Copy Number Variations ; Mutagenesis, Site-Directed ; Polymerase Chain Reaction ; Pyrimethamine/pharmacology ; Tetrahydrofolate Dehydrogenase/genetics ; Tetrahydrofolate Dehydrogenase/metabolism ; Thymidylate Synthase/genetics ; Thymidylate Synthase/metabolism ; Toxoplasma
Chemical Substances	Antigens, Surface ; DNA (9007-49-2) ; Tetrahydrofolate Dehydrogenase (EC 1.5.1.3) ; Thymidylate Synthase (EC 2.1.1.45) ; Pyrimethamine (Z3614QOX8W)
Language	English
Publishing date	2022-09-16
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0271011
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Article: Fatal spirorchiidosis in European pond turtles (

Schönbächler, Katja / Olias, Philipp / Richard, Olivia K / Origgi, Francesco C / Dervas, Eva / Hoby, Stefan / Basso, Walter / Berenguer Veiga, Inês

International journal for parasitology. Parasites and wildlife

2022 Volume 17, Page(s) 144–151

Abstract: Infections with intravascular digenean trematodes of the Spirorchiidae family (spirorchiidoses) are of great conservation concern both in marine and freshwater turtles due to their pathogenic potential. Between 2014 and 2021, ...

Abstract	Infections with intravascular digenean trematodes of the Spirorchiidae family (spirorchiidoses) are of great conservation concern both in marine and freshwater turtles due to their pathogenic potential. Between 2014 and 2021,
Language	English
Publishing date	2022-01-11
Publishing country	England
Document type	Journal Article
ZDB-ID	2715239-X
ISSN	2213-2244
ISSN	2213-2244
DOI	10.1016/j.ijppaw.2022.01.004
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Article: Encephalitis Associated with Sarcocystis halieti Infection in a Free-Ranging Little Owl (Athene noctua)

Maier-Sam, Kristina / Kaiponen, Taina / Schmitz, Anna / Schulze, Christoph / Bock, Sabine / Hlinak, Andreas / Olias, Philipp

Journal of wildlife diseases. 2021 July 5, v. 57, no. 3

2021

Abstract: A juvenile Little Owl (Athene noctua) was diagnosed with granulomatous encephalitis and muscular sarcocysts. Sarcocystis halieti was identified in the brain and muscle tissue by PCR and subsequent sequencing. This is the first report of S. halieti as a ... ...

Abstract	A juvenile Little Owl (Athene noctua) was diagnosed with granulomatous encephalitis and muscular sarcocysts. Sarcocystis halieti was identified in the brain and muscle tissue by PCR and subsequent sequencing. This is the first report of S. halieti as a potential encephalitis-causing pathogen in birds.
Keywords	Athene noctua ; Sarcocystis ; brain ; encephalitis ; juveniles ; muscle tissues ; pathogens ; sarcocysts ; wildlife
Language	English
Dates of publication	2021-0705
Size	p. 712-714.
Publishing place	Wildlife Disease Association
Document type	Article
ZDB-ID	410709-3
ISSN	1943-3700 ; 0090-3558
ISSN (online)	1943-3700
ISSN	0090-3558
DOI	10.7589/JWD-D-20-00184
Database	NAL-Catalogue (AGRICOLA)

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Article: Fatal spirorchiidosis in European pond turtles (Emys orbicularis) in Switzerland

Schönbächler, Katja / Olias, Philipp / Richard, Olivia K. / Origgi, Francesco C. / Dervas, Eva / Hoby, Stefan / Basso, Walter / Berenguer Veiga, Inês

International journal for parasitology. 2022 Apr., v. 17

2022

Abstract	Infections with intravascular digenean trematodes of the Spirorchiidae family (spirorchiidoses) are of great conservation concern both in marine and freshwater turtles due to their pathogenic potential. Between 2014 and 2021, Spirorchis sp. infections associated with granulomatous inflammation and sudden death were detected in European pond turtles (Emys orbicularis) from three conservation breeding facilities in Switzerland. Blood fluke eggs associated with lesions were found in the intestine, spleen, testis, skeletal musculature, heart, kidneys, stomach, pancreas, liver, lung, and meninges from nine pond turtles submitted for necropsy and in the intestinal content from five of these animals. Two novel polymerase chain reactions (PCRs) targeting the 28S ribosomal RNA gene and the ITS2 region and subsequent sequencing revealed 100% nucleotide identity with a Spirorchis sp. previously isolated from an Escambia map turtle (Graptemys ernsti) in the USA. Our findings suggest a spill-over event secondary to direct or indirect contact with invasive North American turtle species in Switzerland. We describe the clinical, haematological, ultrasonographical, endoscopical, parasitological, pathological, and molecular findings associated with spirorchiid blood fluke infections of the Spirorchis genus in E. orbicularis, as well as the biosecurity measures that were developed to prevent the spread of this parasite among breeding and highly endangered free-ranging E. orbicularis populations in Switzerland.
Keywords	Emys orbicularis ; Graptemys ; Spirorchiidae ; biosecurity ; blood flukes ; death ; freshwater ; genes ; heart ; indirect contact ; inflammation ; intestines ; liver ; lungs ; meninges ; necropsy ; pancreas ; parasitology ; pathogenicity ; ribosomal RNA ; spleen ; stomach ; testes ; turtles ; Switzerland
Language	English
Dates of publication	2022-04
Size	p. 144-151.
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	2715239-X
ISSN	2213-2244
ISSN	2213-2244
DOI	10.1016/j.ijppaw.2022.01.004
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Article ; Online: New Haemoproteus parasite of parrots, with remarks on the virulence of haemoproteids in naive avian hosts.

Valkiūnas, Gediminas / Pendl, Helene / Olias, Philipp

Acta tropica

2017 Volume 176, Page(s) 256–262

Abstract: Haemoproteus infections can cause fatal disease in parrots (Psittaciformes), one of the most endangered groups of birds. The great diversity of parrots in tropical and subtropical ecosystems has been markedly understudied in terms of their parasite ... ...

Abstract	Haemoproteus infections can cause fatal disease in parrots (Psittaciformes), one of the most endangered groups of birds. The great diversity of parrots in tropical and subtropical ecosystems has been markedly understudied in terms of their parasite diversity. Only two psittacine Haemoproteus species have been described. Here we report a new Haemoproteus parasite, H. (Parahaemoproteus) homohandai n. sp. (lineage hARCHL01) found in erythrocytes of a Red-and-green macaw Ara chloropterus. We morphologically and genetically characterize the parasite based on a segment of the mitochondrial cytochrome b gene, which can be used for identification and diagnosis of infection. This is the first Haemoproteus species described from South American parrots and the first genetically characterized psittacine Haemoproteus sp. Haemoproteus homohandai n. sp. can be readily distinguished from other haemoproteids by its growing circumnuclear and close to circumnuclear macrogametocytes, which are strictly associated with erythrocyte nuclei, but do not touch the erythrocyte envelope along their entire margin and do not fill erythrocytes up to their poles. Illustrations of growing and mature gametocytes of the new species are given, and a phylogenetic analysis identifies the position of this parasite lineage in relation to other Haemoproteus parasites. Importantly, H. homohandai n. sp. and all other Haemoproteus lineages reported from parrots cluster with species of the subgenus Parahaemoproteus, indicating the transmission by Culicoides biting midges.
Language	English
Publishing date	2017-12
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	210415-5
ISSN	1873-6254 ; 0001-706X
ISSN (online)	1873-6254
ISSN	0001-706X
DOI	10.1016/j.actatropica.2017.08.004
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Article ; Online: A parasite DNA binding protein with potential to influence disease susceptibility acts as an analogue of mammalian HMGA transcription factors.

Durrani, Zeeshan / Kinnaird, Jane / Cheng, Chew Weng / Brühlmann, Francis / Capewell, Paul / Jackson, Andrew / Larcombe, Stephen / Olias, Philipp / Weir, William / Shiels, Brian

PloS one

2023 Volume 18, Issue 6, Page(s) e0286526

Abstract: Intracellular pathogens construct their environmental niche, and influence disease susceptibility, by deploying factors that manipulate infected host cell gene expression. Theileria annulata is an important tick-borne parasite of cattle that causes ... ...

Abstract	Intracellular pathogens construct their environmental niche, and influence disease susceptibility, by deploying factors that manipulate infected host cell gene expression. Theileria annulata is an important tick-borne parasite of cattle that causes tropical theileriosis. Excellent candidates for modulating host cell gene expression are DNA binding proteins bearing AT-hook motifs encoded within the TashAT gene cluster of the parasite genome. In this study, TashAT2 was transfected into bovine BoMac cells to generate three expressing and three non-expressing (opposite orientation) cell lines. RNA-Seq was conducted and differentially expressed (DE) genes identified. The resulting dataset was compared with genes differentially expressed between infected cells and non-infected cells, and DE genes between infected cell lines from susceptible Holstein vs tolerant Sahiwal cattle. Over 800 bovine genes displayed differential expression associated with TashAT2, 209 of which were also modulated by parasite infection. Network analysis showed enrichment of DE genes in pathways associated with cellular adhesion, oncogenesis and developmental regulation by mammalian AT-hook bearing high mobility group A (HMGA) proteins. Overlap of TashAT2 DE genes with Sahiwal vs Holstein DE genes revealed that a significant number of shared genes were associated with disease susceptibility. Altered protein levels encoded by one of these genes (GULP1) was strongly linked to expression of TashAT2 in BoMac cells and was demonstrated to be higher in infected Holstein leucocytes compared to Sahiwal. We conclude that TashAT2 operates as an HMGA analogue to differentially mould the epigenome of the infected cell and influence disease susceptibility.
MeSH term(s)	Cattle ; Animals ; DNA-Binding Proteins/genetics ; Disease Susceptibility ; Transcription Factors/metabolism ; Parasites/metabolism ; Theileriasis/parasitology ; Theileria annulata/genetics ; HMGA Proteins/metabolism ; Mammals/metabolism
Chemical Substances	DNA-Binding Proteins ; Transcription Factors ; HMGA Proteins
Language	English
Publishing date	2023-06-05
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0286526
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Article ; Online: Toxoplasma

Hakimi, Mohamed-Ali / Olias, Philipp / Sibley, L David

Clinical microbiology reviews

2017 Volume 30, Issue 3, Page(s) 615–645

Abstract: Early electron microscopy studies revealed the elaborate cellular features that define the unique adaptations of apicomplexan parasites. Among these were bulbous rhoptry (ROP) organelles and small, dense granules (GRAs), both of which are secreted during ...

Abstract	Early electron microscopy studies revealed the elaborate cellular features that define the unique adaptations of apicomplexan parasites. Among these were bulbous rhoptry (ROP) organelles and small, dense granules (GRAs), both of which are secreted during invasion of host cells. These early morphological studies were followed by the exploration of the cellular contents of these secretory organelles, revealing them to be comprised of highly divergent protein families with few conserved domains or predicted functions. In parallel, studies on host-pathogen interactions identified many host signaling pathways that were mysteriously altered by infection. It was only with the advent of forward and reverse genetic strategies that the connections between individual parasite effectors and the specific host pathways that they targeted finally became clear. The current repertoire of parasite effectors includes ROP kinases and pseudokinases that are secreted during invasion and that block host immune pathways. Similarly, many secretory GRA proteins alter host gene expression by activating host transcription factors, through modification of chromatin, or by inducing small noncoding RNAs. These effectors highlight novel mechanisms by which
MeSH term(s)	Animals ; Host-Parasite Interactions/immunology ; Host-Parasite Interactions/physiology ; Humans ; Signal Transduction ; Toxoplasma/genetics ; Toxoplasma/physiology ; Transcription, Genetic
Language	English
Publishing date	2017
Publishing country	United States
Document type	Journal Article ; Review ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	645015-5
ISSN	1098-6618 ; 0893-8512
ISSN (online)	1098-6618
ISSN	0893-8512
DOI	10.1128/CMR.00005-17
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