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Article ; Online: Effects of host species on microbiota composition in Phlebotomus and Lutzomyia sand flies.

Tabbabi, Ahmed / Mizushima, Daiki / Yamamoto, Daisuke S / Kato, Hirotomo

2023 Volume 16, Issue 1, Page(s) 310

Abstract: Background: Blood-sucking phlebotomine sand flies are vectors of the protozoan parasites Leishmania spp. Although the intestinal microbiota is involved in a wide range of biological and physiological processes and has the potential to alter vector ... ...

Abstract	Background: Blood-sucking phlebotomine sand flies are vectors of the protozoan parasites Leishmania spp. Although the intestinal microbiota is involved in a wide range of biological and physiological processes and has the potential to alter vector competence, little is known about the factors that modify the gut microbiota composition of sand flies. As a key step toward addressing this issue, we investigated the impact of host species on the gut bacterial composition in Phlebotomus and Lutzomyia sand flies reared under the same conditions. Methods: Bacterial 16S rRNA gene amplification and Illumina MiSeq sequencing were used to characterize the overall bacterial composition of three laboratory-reared sandflies: Phlebotomus papatasi, Ph. duboscqi, and Lutzomyia longipalpis. Results: Our results showed that the larvae of the three sand fly species harbored almost the same microbes but had different relative abundances. Adult Ph. papatasi and Ph. duboscqi revealed similar microbiome compositions, which were distinct from that of adult Lu. longipalpis. Furthermore, we showed that Ph. papatasi and Ph. duboscqi are hosts for different bacterial genera. The experiment was repeated twice to improve accuracy and increase reliability of the data, and the same results were obtained even when a distinct composition of the microbiome among the same species was identified probably because of the use of different larvae food batch. Conclusions: The present study provides key insights into the role of host species in the gut microbial content of different sand fly species reared under the same conditions, which may influence their susceptibility to Leishmania infection.
MeSH term(s)	Animals ; Phlebotomus ; Psychodidae ; RNA, Ribosomal, 16S/genetics ; Reproducibility of Results ; Microbiota ; Larva
Chemical Substances	RNA, Ribosomal, 16S
Language	English
Publishing date	2023-08-31
Publishing country	England
Document type	Journal Article
ZDB-ID	2409480-8
ISSN	1756-3305 ; 1756-3305
ISSN (online)	1756-3305
ISSN	1756-3305
DOI	10.1186/s13071-023-05939-2
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Article ; Online: A real-time PCR for quantification of parasite burden and its correlations with clinical characteristics and anti-rKRP42 IgG level in cutaneous leishmaniasis in Sri Lanka.

De Silva, Nirmitha Lalindi / De Silva, Viraji Nefertiti Hiromel / Weerasooriya, Mirani Vasanthamala / Takagi, Hidekazu / Itoh, Makoto / Kato, Hirotomo / Yahathugoda, Thishan Channa

Parasitology international

2024 Volume 100, Page(s) 102865

Abstract: In visceral and mucocutaneous leishmaniasis, humoral immune response can reflect disease severity and parasite burden. Cutaneous leishmaniasis (CL) in Sri Lanka is caused by a usually visceralizing parasite, Leishmania donovani. We assessed the parasite ... ...

Abstract	In visceral and mucocutaneous leishmaniasis, humoral immune response can reflect disease severity and parasite burden. Cutaneous leishmaniasis (CL) in Sri Lanka is caused by a usually visceralizing parasite, Leishmania donovani. We assessed the parasite burden (relative quantity-RQ) in 190 CL patients using quantitative real-time PCR (qPCR-with primers designed for this study) and smear microscopy, then correlated it with clinical parameters and IgG response. RQ of parasite DNA was determined with human-specific glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as the internal control. The qPCR sensitivity was tested with serially diluted DNA from cultured L. donovani parasites. Smears were assigned a score based on number of parasites per high power field. Data from previous studies were used for comparison and correlation; nested Internal Transcribed Spacer 1 (ITS1) PCR as reference standard (RS) and IgG antibody titers to the Leishmania rKRp42 antigen as the immune response. The qPCR amplified and quantified 86.8% of the samples while demonstrating a fair and significant agreement with ITS1-PCR and microscopy. Parasite burden by qPCR and microscopy were highly correlated (r = 0.76; p = 0.01) but showed no correlation with the IgG response (r = 0.056; p = 0.48). Corresponding mean RQs of IgG titers grouped by percentiles, showed no significant difference (p = 0.93). Mean RQ was higher in early lesions (p = 0.04), decreased with lesion size (p = 0.12) and slightly higher among papules, nodules and wet ulcers (p = 0.72). Our study established qPCR's efficacy in quantifying parasite burden in Sri Lankan CL lesions but no significant correlation was observed between the parasite burden and host IgG response to the Leishmania rKRP42 antigen.
MeSH term(s)	Animals ; Humans ; Parasites ; Real-Time Polymerase Chain Reaction ; Sri Lanka/epidemiology ; Leishmaniasis, Cutaneous/epidemiology ; Leishmaniasis, Cutaneous/parasitology ; Leishmania donovani/genetics ; DNA ; Immunoglobulin G
Chemical Substances	DNA (9007-49-2) ; Immunoglobulin G
Language	English
Publishing date	2024-02-08
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	1363151-2
ISSN	1873-0329 ; 1383-5769
ISSN (online)	1873-0329
ISSN	1383-5769
DOI	10.1016/j.parint.2024.102865
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Article ; Online: A rare sugar, allose, inhibits the development of

Mizushima, Daiki / Yamamoto, Daisuke S / Tabbabi, Ahmed / Arai, Meiji / Kato, Hirotomo

Frontiers in cellular and infection microbiology

2023 Volume 13, Page(s) 1162918

Abstract: A rare sugar, allose, was reported to inhibit the development ... ...

Abstract	A rare sugar, allose, was reported to inhibit the development of
MeSH term(s)	Animals ; Anopheles/parasitology ; Parasites ; Sugars ; Mosquito Vectors ; Carbohydrates ; Plasmodium berghei ; Malaria/parasitology ; Bacteria ; Microbiota
Chemical Substances	allose (6038-51-3) ; Sugars ; Carbohydrates
Language	English
Publishing date	2023-07-20
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2619676-1
ISSN	2235-2988 ; 2235-2988
ISSN (online)	2235-2988
ISSN	2235-2988
DOI	10.3389/fcimb.2023.1162918
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Article: Ayaconin, a novel inhibitor of the plasma contact system from the sand fly Lutzomyia ayacuchensis, a vector of Andean-type cutaneous leishmaniasis

Kawahori, Satoru / Seki, Chisato / Mizushima, Daiki / Tabbabi, Ahmed / Yamamoto, Daisuke S. / Kato, Hirotomo

Acta tropica. 2022 Oct., v. 234

2022

Abstract: Transcriptome analysis of the salivary gland cDNA library from a phlebotomine sand fly, Lutzomyia ayacuchensis, identified a transcript coding for the PpSP15/SL1 family protein as the second most abundant salivary component. In the present study, a ... ...

Abstract	Transcriptome analysis of the salivary gland cDNA library from a phlebotomine sand fly, Lutzomyia ayacuchensis, identified a transcript coding for the PpSP15/SL1 family protein as the second most abundant salivary component. In the present study, a recombinant protein of the PpSP15/SL1 family protein, designated ayaconin, was expressed in Escherichia coli, and its biological activity was characterized. The recombinant ayaconin purified from the soluble fraction of E. coli lysate efficiently inhibited the intrinsic but not extrinsic blood coagulation pathway. When the target of ayaconin was evaluated using fluorescent substrates of coagulation factors, ayaconin inhibited factor XIIa (FXIIa) activity more efficiently in a dose-dependent manner, suggesting that FXII is the primary target of ayaconin. In addition, incubation of ayaconin with FXII prior to activation effectively inhibited FXIIa activity, whereas such inhibition was not observed when ayaconin was mixed after the production of FXIIa, indicating that ayaconin inhibits the activation process of FXII to produce FXIIa, but not the enzymatic activity of FXIIa. Moreover, ayaconin was shown to bind to FXII, suggesting that the binding of ayaconin to FXII is involved in the inhibitory mechanism against FXII activation. These results suggest that ayaconin plays an important role in the blood-sucking of Lu. ayacuchensis.
Keywords	Escherichia coli ; Lutzomyia ayacuchensis ; bioactive properties ; blood coagulation ; cDNA libraries ; coagulation ; cutaneous leishmaniasis ; dose response ; enzyme activity ; fluorescence ; recombinant proteins ; salivary glands ; transcriptomics
Language	English
Dates of publication	2022-10
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	210415-5
ISSN	1873-6254 ; 0001-706X
ISSN (online)	1873-6254
ISSN	0001-706X
DOI	10.1016/j.actatropica.2022.106602
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Article ; Online: Case Report: Successful Treatment with Miltefosine of Severe New World Mucosal Leishmaniasis Caused by

Calvopina, Manuel / Jijon, Sara / Serrano, Esteban / Kato, Hirotomo

The American journal of tropical medicine and hygiene

2020 Volume 103, Issue 2, Page(s) 752–755

Abstract: An 88-year-old man with mutilating mucosal leishmaniasis (ML) involving septal perforation, with granulomas in the pharynx and larynx, was treated with oral miltefosine, 50 mg three times/day for 28 days. Miltefosine, an antineoplastic agent, is ... ...

Abstract	An 88-year-old man with mutilating mucosal leishmaniasis (ML) involving septal perforation, with granulomas in the pharynx and larynx, was treated with oral miltefosine, 50 mg three times/day for 28 days. Miltefosine, an antineoplastic agent, is considered an alternative option for the treatment of ML, showing efficacies of 75-92% in Bolivia, Brazil, and Argentina. The patient denied having previous cutaneous (CL) leishmaniasis, and no CL lesions were recognized by physical examination. Parasites obtained from mucosal lesions were identified by cytochrome
MeSH term(s)	Aged, 80 and over ; Antiprotozoal Agents/therapeutic use ; Cytochromes b/genetics ; Dysphonia/etiology ; Humans ; Leishmania guyanensis/genetics ; Leishmania guyanensis/isolation & purification ; Leishmaniasis, Mucocutaneous/drug therapy ; Male ; Nasal Septal Perforation/etiology ; Nose Diseases/complications ; Nose Diseases/drug therapy ; Nose Diseases/pathology ; Pharyngeal Diseases/complications ; Pharyngeal Diseases/drug therapy ; Pharyngeal Diseases/pathology ; Phosphorylcholine/analogs & derivatives ; Phosphorylcholine/therapeutic use ; Severity of Illness Index
Chemical Substances	Antiprotozoal Agents ; Phosphorylcholine (107-73-3) ; miltefosine (53EY29W7EC) ; Cytochromes b (9035-37-4)
Language	English
Publishing date	2020-06-04
Publishing country	United States
Document type	Case Reports ; Journal Article
ZDB-ID	2942-7
ISSN	1476-1645 ; 0002-9637
ISSN (online)	1476-1645
ISSN	0002-9637
DOI	10.4269/ajtmh.19-0867
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Article ; Online: Ayaconin, a novel inhibitor of the plasma contact system from the sand fly Lutzomyia ayacuchensis, a vector of Andean-type cutaneous leishmaniasis.

Kawahori, Satoru / Seki, Chisato / Mizushima, Daiki / Tabbabi, Ahmed / Yamamoto, Daisuke S / Kato, Hirotomo

Acta tropica

2022 Volume 234, Page(s) 106602

Abstract	Transcriptome analysis of the salivary gland cDNA library from a phlebotomine sand fly, Lutzomyia ayacuchensis, identified a transcript coding for the PpSP15/SL1 family protein as the second most abundant salivary component. In the present study, a recombinant protein of the PpSP15/SL1 family protein, designated ayaconin, was expressed in Escherichia coli, and its biological activity was characterized. The recombinant ayaconin purified from the soluble fraction of E. coli lysate efficiently inhibited the intrinsic but not extrinsic blood coagulation pathway. When the target of ayaconin was evaluated using fluorescent substrates of coagulation factors, ayaconin inhibited factor XIIa (FXIIa) activity more efficiently in a dose-dependent manner, suggesting that FXII is the primary target of ayaconin. In addition, incubation of ayaconin with FXII prior to activation effectively inhibited FXIIa activity, whereas such inhibition was not observed when ayaconin was mixed after the production of FXIIa, indicating that ayaconin inhibits the activation process of FXII to produce FXIIa, but not the enzymatic activity of FXIIa. Moreover, ayaconin was shown to bind to FXII, suggesting that the binding of ayaconin to FXII is involved in the inhibitory mechanism against FXII activation. These results suggest that ayaconin plays an important role in the blood-sucking of Lu. ayacuchensis.
MeSH term(s)	Animals ; Escherichia coli/genetics ; Factor XIIa/metabolism ; Insect Vectors ; Leishmaniasis, Cutaneous ; Phlebotomus ; Psychodidae/genetics
Chemical Substances	Factor XIIa (EC 3.4.21.38)
Language	English
Publishing date	2022-07-08
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.2022.106602
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Article: A prospective mechanism and source of cholesterol uptake by Plasmodium falciparum-infected erythrocytes co-cultured with HepG2 cells

Hayakawa, Eri H / Kato, Hirotomo / Nardone, Glenn A / Usukura, Jiro

Elsevier B.V. Parasitology international. 2021 Feb., v. 80

2021

Abstract: Plasmodium falciparum (P. falciparum) parasites still cause lethal infections worldwide, especially in Africa (https://www.who.int/publications/i/item/world-malaria-report-2019). During P. falciparum blood-stage infections in humans, low-density ... ...

Abstract	Plasmodium falciparum (P. falciparum) parasites still cause lethal infections worldwide, especially in Africa (https://www.who.int/publications/i/item/world-malaria-report-2019). During P. falciparum blood-stage infections in humans, low-density lipoprotein, high-density lipoprotein and cholesterol levels in the blood become low. Because P. falciparum lacks a de novo cholesterol synthesis pathway, it must import cholesterol from the surrounding environment. However, the origin of the cholesterol and how it is taken up by the parasite across the multiple membranes that surround it is not fully understood. To answer this, we used a cholesterol synthesis inhibiter (simvastatin), a cholesterol transport inhibitor (ezetimibe), and an activating ligand of the peroxisome proliferator-activated receptor α, called ciprofibrate, to investigate the effects of these agents on the intraerythrocytic growth of P. falciparum, both with and without HepG2 cells as the lipoprotein feeders. P. falciparum growth was inhibited in the presence of ezetimibe, but ezetimibe was not very effective at inhibiting P. falciparum growth when used in the co-culture system, unlike simvastatin, which strongly promoted parasite growth in this system. Ezetimibe is known to inhibit cholesterol absorption by blocking the activity of Niemann-Pick C1 like 1 (NPC1L1) protein, and simvastatin is known to enhance NPC1L1 expression in the human body's small intestine. Collectively, our results support the possibility that cholesterol import by P. falciparum involves hepatocytes, and cholesterol uptake into the parasite occurs via NPC1L1 protein or an NPC1L1 homolog during the erythrocytic stages of the P. falciparum lifecycle.
Keywords	Plasmodium falciparum ; absorption ; cholesterol ; coculture ; environment ; erythrocytes ; hepatocytes ; high density lipoprotein ; human cell lines ; humans ; ligands ; low density lipoprotein ; parasites ; parasitology ; peroxisome proliferator-activated receptors ; small intestine ; synthesis ; Africa
Language	English
Dates of publication	2021-02
Publishing place	Elsevier B.V.
Document type	Article
Note	NAL-light
ZDB-ID	1363151-2
ISSN	1383-5769
ISSN	1383-5769
DOI	10.1016/j.parint.2020.102179
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Article ; Online: Loop-mediated isothermal amplification (LAMP): An advanced molecular point-of-care technique for the detection of Leishmania infection.

Nzelu, Chukwunonso O / Kato, Hirotomo / Peters, Nathan C

PLoS neglected tropical diseases

2019 Volume 13, Issue 11, Page(s) e0007698

Abstract: Leishmaniasis, caused by protozoan parasites of the Leishmania genus, represents an important health problem in many regions of the world. Lack of effective point-of-care (POC) diagnostic tests applicable in resources-limited endemic areas is a critical ... ...

Abstract	Leishmaniasis, caused by protozoan parasites of the Leishmania genus, represents an important health problem in many regions of the world. Lack of effective point-of-care (POC) diagnostic tests applicable in resources-limited endemic areas is a critical barrier to effective treatment and control of leishmaniasis. The development of the loop-mediated isothermal amplification (LAMP) assay has provided a new tool towards the development of a POC diagnostic test based on the amplification of pathogen DNA. LAMP does not require a thermocycler, is relatively inexpensive, and is simple to perform with high amplification sensitivity and specificity. In this review, we discuss the current technical developments, applications, diagnostic performance, challenges, and future of LAMP for molecular diagnosis and surveillance of Leishmania parasites. Studies employing the LAMP assay to diagnose human leishmaniasis have reported sensitivities of 80% to 100% and specificities of 94% to 100%. These observations suggest that LAMP offers a good molecular POC technique for the diagnosis of leishmaniasis and is also readily applicable to screening at-risk populations and vector sand flies for Leishmania infection in endemic areas.
MeSH term(s)	Animals ; DNA, Protozoan/genetics ; Databases, Factual ; Dog Diseases/diagnosis ; Dog Diseases/parasitology ; Dogs ; Humans ; Leishmania/genetics ; Leishmaniasis/diagnosis ; Mass Screening/methods ; Molecular Diagnostic Techniques/methods ; Nucleic Acid Amplification Techniques/economics ; Nucleic Acid Amplification Techniques/methods ; Point-of-Care Systems ; Psychodidae/parasitology ; Sensitivity and Specificity ; Time Factors
Chemical Substances	DNA, Protozoan
Language	English
Publishing date	2019-11-07
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2429704-5
ISSN	1935-2735 ; 1935-2727
ISSN (online)	1935-2735
ISSN	1935-2727
DOI	10.1371/journal.pntd.0007698
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Article ; Online: A prospective mechanism and source of cholesterol uptake by Plasmodium falciparum-infected erythrocytes co-cultured with HepG2 cells.

Hayakawa, Eri H / Kato, Hirotomo / Nardone, Glenn A / Usukura, Jiro

Parasitology international

2020 Volume 80, Page(s) 102179

Abstract	Plasmodium falciparum (P. falciparum) parasites still cause lethal infections worldwide, especially in Africa (https://www.who.int/publications/i/item/world-malaria-report-2019). During P. falciparum blood-stage infections in humans, low-density lipoprotein, high-density lipoprotein and cholesterol levels in the blood become low. Because P. falciparum lacks a de novo cholesterol synthesis pathway, it must import cholesterol from the surrounding environment. However, the origin of the cholesterol and how it is taken up by the parasite across the multiple membranes that surround it is not fully understood. To answer this, we used a cholesterol synthesis inhibiter (simvastatin), a cholesterol transport inhibitor (ezetimibe), and an activating ligand of the peroxisome proliferator-activated receptor α, called ciprofibrate, to investigate the effects of these agents on the intraerythrocytic growth of P. falciparum, both with and without HepG2 cells as the lipoprotein feeders. P. falciparum growth was inhibited in the presence of ezetimibe, but ezetimibe was not very effective at inhibiting P. falciparum growth when used in the co-culture system, unlike simvastatin, which strongly promoted parasite growth in this system. Ezetimibe is known to inhibit cholesterol absorption by blocking the activity of Niemann-Pick C1 like 1 (NPC1L1) protein, and simvastatin is known to enhance NPC1L1 expression in the human body's small intestine. Collectively, our results support the possibility that cholesterol import by P. falciparum involves hepatocytes, and cholesterol uptake into the parasite occurs via NPC1L1 protein or an NPC1L1 homolog during the erythrocytic stages of the P. falciparum lifecycle.
MeSH term(s)	Anticholesteremic Agents/pharmacology ; Cholesterol/metabolism ; Erythrocytes/metabolism ; Ezetimibe/pharmacology ; Fibric Acids/pharmacology ; Hep G2 Cells ; Humans ; Hypolipidemic Agents/pharmacology ; Plasmodium falciparum/physiology ; Simvastatin/pharmacology
Chemical Substances	Anticholesteremic Agents ; Fibric Acids ; Hypolipidemic Agents ; Cholesterol (97C5T2UQ7J) ; Simvastatin (AGG2FN16EV) ; Ezetimibe (EOR26LQQ24) ; ciprofibrate (F8252JGO9S)
Language	English
Publishing date	2020-08-25
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	1363151-2
ISSN	1873-0329 ; 1383-5769
ISSN (online)	1873-0329
ISSN	1383-5769
DOI	10.1016/j.parint.2020.102179
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Article ; Online: Natural infections of Pintomyia verrucarum and Pintomyia maranonensis by Leishmania (Viannia) peruviana in the Eastern Andes of northern Peru.

Kato, Hirotomo / Seki, Chisato / Kubo, Makoto / Gonzales-Cornejo, Lizandro / Caceres, Abraham G

PLoS neglected tropical diseases

2021 Volume 15, Issue 4, Page(s) e0009352

Abstract: The natural infection of sand flies by Leishmania was investigated in Andean areas located between the Central and Eastern Cordilleras of northern Peru where cutaneous leishmaniasis caused by Leishmania (Viannia) peruviana is endemic. Sand flies were ... ...

Abstract	The natural infection of sand flies by Leishmania was investigated in Andean areas located between the Central and Eastern Cordilleras of northern Peru where cutaneous leishmaniasis caused by Leishmania (Viannia) peruviana is endemic. Sand flies were captured at five locations along the Utcubamba River in the Department of Amazonas, and morphologically identified under a microscope. Among 422 female sand flies dissected, the most dominant species was Pintomyia verrucarum (320 flies), followed by Pi. maranonensis (83 flies), Pi. robusta (13 flies), and Lutzomyia castanea (6 flies). Genetic analysis of sand flies from these areas together with those from other areas revealed that individuals of Pi. verrucarum were closely related regardless of morphological variation of their spermathecae. On the other hand, individuals of Pi. maranonensis collected in the study area were distant from those of other areas with genetic distances over the intraspecific level but mostly below the interspecific level, suggesting the unique characteristics of sand flies in this area. The natural infection of sand flies by flagellate parasites was detected mainly in the hindgut of each one of Pi. verrucarum and Pi. maranonensis. Both parasite species were identified as L. (V.) peruviana based on cytochrome b and mannose phosphate isomerase gene analyses. In addition, parasite species obtained from the lesion of a patient with cutaneous leishmaniasis in the study area in this period was identified as L. (V.) peruviana. These results strongly suggest that Pi. verrucarum and Pi. maranonensis are responsible for the transmission of L. (V.) peruviana in these areas. This is the first report of the natural infection of Pi. maranonensis by L. (V.) peruviana.
Language	English
Publishing date	2021-04-15
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2429704-5
ISSN	1935-2735 ; 1935-2727
ISSN (online)	1935-2735
ISSN	1935-2727
DOI	10.1371/journal.pntd.0009352
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