Publications by authors named "Noboru Inoue"

173 Publications

In Vitro and In Vivo Trypanocidal Efficacy of Synthesized Nitrofurantoin Analogs.

Molecules 2021 Jun 2;26(11). Epub 2021 Jun 2.

Research Center for Global Agromedicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.

African trypanosomes cause diseases in humans and livestock. Human African trypanosomiasis is caused by and . Animal trypanosomoses have major effects on livestock production and the economy in developing countries, with disease management depending mainly on chemotherapy. Moreover, only few drugs are available and these have adverse effects on patients, are costly, show poor accessibility, and parasites develop drug resistance to them. Therefore, novel trypanocidal drugs are urgently needed. Here, the effects of synthesized nitrofurantoin analogs were evaluated against six species/strains of animal and human trypanosomes, and the treatment efficacy of the selected compounds was assessed in vivo. Analogs and , containing 11- and 12-carbon aliphatic chains, respectively, showed the highest trypanocidal activity (IC < 0.34 µM) and the lowest cytotoxicity (IC > 246.02 µM) in vitro. Structure-activity relationship analysis suggested that the trypanocidal activity and cytotoxicity were related to the number of carbons in the aliphatic chain and electronegativity. In vivo experiments, involving oral treatment with nitrofurantoin, showed partial efficacy, whereas the selected analogs showed no treatment efficacy. These results indicate that nitrofurantoin analogs with high hydrophilicity are required for in vivo assessment to determine if they are promising leads for developing trypanocidal drugs.
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http://dx.doi.org/10.3390/molecules26113372DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8199755PMC
June 2021

Synthesis and evaluation of trypanocidal activity of derivatives of naturally occurring 2,5-diphenyloxazoles.

Bioorg Med Chem 2021 Jul 12;42:116253. Epub 2021 Jun 12.

Faculty of Pharmaceutical Sciences, Tohoku Medical and Pharmaceutical University, 4-4-1 Komatsushima, Aoba-ku, Sendai 981-8558, Japan.

African trypanosomiasis is a zoonotic protozoan disease affecting the nervous system. Various natural products reportedly exhibit trypanocidal activity. Naturally occurring 2,5-diphenyloxazoles present in Oxytropis lanata, and their derivatives, were synthesized. The trypanocidal activities of the synthesized compounds were evaluated against Trypanosoma brucei brucei, T. b. gambiense, T. b. rhodesiense, T. congolense, and T. evansi. Natural product 1 exhibited trypanocidal activity against all the species/subspecies of trypanosomes, exhibiting half-maximal inhibitory concentrations (IC) of 1.1-13.5 μM. Modification of the oxazole core improved the trypanocidal activity. The 1,3,4-oxadiazole (7) and 2,4-diphenyloxazole (9) analogs exhibited potency superior to that of 1. However, these compounds exhibited cytotoxicity in Madin-Darby bovine kidney cells. The O-methylated analog of 1 (12) was non-cytotoxic and exhibited selective trypanocidal activity against T. congolense (IC = 0.78 µM). Structure-activity relationship studies of the 2,5-diphenyloxazole analogs revealed aspects of the molecular structure critical for maintaining selective trypanocidal activity against T. congolense.
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http://dx.doi.org/10.1016/j.bmc.2021.116253DOI Listing
July 2021

First molecular detection and identification of Trypanosoma evansi in goats from Cebu, Philippines using a PCR-based assay.

Vet Parasitol Reg Stud Reports 2020 07 19;21:100414. Epub 2020 May 19.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido 080-8555, Japan; Research Center for Global Agromedicine, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido 080-8555, Japan. Electronic address:

The increasing number and severity of surra outbreaks in the Philippines led the government to consider it as the second most important disease of livestock in the country. It is one of the most economically important animal parasitic diseases and has been reported in several animal species, including water buffaloes, cattle, and horses in different regions of the Philippines. However, it has not yet been reported in Cebu, the usual gateway of livestock trade in the area that raises 6% of the 3.75 million goats in the country. In the current study, a PCR-based assay was conducted for the molecular detection and characterization of Trypanosoma evansi in goats in Cebu. A total of 251 goats were randomly sampled from four farms. DNA was extracted and ITS1-PCR was applied to detect different trypanosomes in goats. Eighty-five out of the 251 (33.9%) samples tested positive for T. evansi, two of which were also positive for T. theileri-like trypanosome. The detection rate of T. evansi was slightly higher in male goats (38.3%) than in females (32.5%), and in younger goats (34.5%) than in adults (33.5%). The findings, however, did not differ significantly to suggest any association between sex and age with T. evansi infection in goats. The detection of T. evansi and T. theileri-like trypanosome in goats was confirmed by sequence analysis of ITS1 region. To our knowledge, this is the first report on the molecular detection and identification of caprine T. evansi infection in Cebu, Philippines.
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http://dx.doi.org/10.1016/j.vprsr.2020.100414DOI Listing
July 2020

Immunohistochemical phenotyping of macrophages and T lymphocytes infiltrating in peripheral nerve lesions of dourine-affected horses.

J Vet Med Sci 2020 Oct 12;82(10):1502-1505. Epub 2020 Aug 12.

Research Center for Global Agromedicine, Obihiro University of Agriculture and Veterinary Medicine, Nishi 2-11, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.

Dourine is a deadly protozoan disease in equids caused by infection with Trypanosoma equiperdum. Neurological signs in the later stage of infection may be caused by peripheral polyneuritis and related axonal degeneration. This neuritis involves T lymphocytes, B lymphocytes, and macrophages, and is observed in cases without obvious neurological signs. However, the pathogenesis of neuritis remains unclear. We identified M2 macrophages and CD8 T cells as the predominant phenotypes in neuritis of dourine-affected horses with or without neurological signs. In contrast, the populations of M1 macrophages and CD4 T cells were small. This result indicates that inflammation was chronic and suggests that dourine-associated neuritis occurs at the early stage of infection.
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http://dx.doi.org/10.1292/jvms.20-0172DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7653314PMC
October 2020

Nationwide serological surveillance of non-tsetse-transmitted horse trypanosomoses in Mongolia.

Parasite Epidemiol Control 2020 Aug 25;10:e00158. Epub 2020 Jun 25.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido 080-8555, Japan.

In Mongolia, horses play important roles, not only in livestock production, but also in terms of culture, tradition, and Mongolian beliefs. Although the presence of non-tsetse-transmitted horse trypanosomoses, which are caused by infections with (surra) and (dourine), has been reported in the country, whether there is a nationwide epidemic of these infectious diseases is unknown. In the present study, a nationwide surveillance of horse trypanosomoses was performed. The sample sizes for each province, the whole country, and male and female horses were, respectively, 96, 2,400, and 316 and 306. In total, 3,641 samples of horse sera were collected by simple random sampling. The rTeGM6-4r-based ELISA, which was applied for surra against cattle and water buffalo and dourine against horse, revealed that the overall sero-prevalence of the diseases in Mongolia was 4.8%. Among them, high sero-prevalences were observed in the central provinces (5.2-11.0%,  < 0.05) of the country. The sero-prevalence was significantly higher in females than in males (6.0% and 4.0%,  < 0.05, respectively) and in non-castrated males (8.4%,  < 0.01) compared with castrated males (3.0%). These results suggested that currently, horse trypanosomoses are a nationwide endemic problem in Mongolia. Knowledge of the nationwide endemic status of non-tsetse-transmitted horse trypanosomoses in Mongolia will be useful to prevent these diseases.
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http://dx.doi.org/10.1016/j.parepi.2020.e00158DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7334808PMC
August 2020

Treatment Efficiency of Combination Therapy With Diminazene Aceturate and Quinapyramine Sulfate in a Horse With Dourine.

J Equine Vet Sci 2020 04 2;87:102905. Epub 2020 Jan 2.

OIE Reference Laboratory for Surra, National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan; Research Center for Global Agromedicine, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan. Electronic address:

Dourine is a lethal protozoan disease of equids, and it is caused by Trypanosoma equiperdum infection via coitus. To date, treatment strategies against the dourine are not recommended because of the frequent relapses; therefore, the World Organisation for Animal Health recommends the stamping-out policy for the control of dourine. Our previous studies have revealed a number of horses with dourine in Mongolia that is the fifth largest horse-breeding country. It is difficult to apply the stamping-out policy for cases of dourine in Mongolia because of an inadequate livestock guarantee system. Therefore, the development of effective treatment measures is an urgent need. In this study, an 8-year-old stallion was definitely diagnosed with dourine based on clinical signs, molecular analysis, and microscopic examination of trypanosomes. Combination therapy with diminazene aceturate and quinapyramine sulfate was applied. Before the treatment, the characteristic clinical signs of dourine were observed, and trypanosomes were detected in the urogenital tract mucosal swab samples by microscopic examination and polymerase chain reaction (PCR). Moreover, positive serological results were obtained. After the treatment, we observed an improvement in the health of the treated horse and no trypanosome infection in its urogenital tract by microscopic examination and PCR. Moreover, serological tests showed seronegative results. The horse has showed no relapse for at least 2.5 years after the treatment, and its reproductive ability has improved. Our result suggests that trypanosomes did not invade cerebrospinal fluid when we started the therapy. In conclusion, the combination therapy has therapeutic potential against dourine at an early phase.
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http://dx.doi.org/10.1016/j.jevs.2019.102905DOI Listing
April 2020

Serological and molecular detection of selected hemoprotozoan parasites in donkeys in West Omdurman, Khartoum State, Sudan.

J Vet Med Sci 2020 Mar 22;82(3):286-293. Epub 2020 Jan 22.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido 080-8555, Japan.

In Sudan, donkeys are important animals, providing transportation and income possibilities. However, the prevalence of parasites in donkeys in Sudan has not been thoroughly characterized. Accordingly, in this study, we aimed to detect selected hemoprotozoan parasites in donkeys in West Omdurman, Khartoum State, Sudan, wherein people depend mainly on donkeys for their daily life. In total, 198 blood samples collected from donkeys in a local market in West Omdurman, were screened using serological and molecular diagnostic techniques. Serologically, 52 (26.3%), 56 (28.3%), and 19 (9.6%) samples were positive for trypanosomosis using Card Agglutination Test for Trypanosoma evansi, Trypanosoma evansi crude antigen -based enzyme-linked immuno sorbent assay (ELISA) and recombinant Trypanosoma evansi GM6-4r-based ELISA, respectively. ELISA for equine piroplasmosis revealed 156 (78.8%) and 10 (5.1%)Theileria equi- and Babesia caballi-positive samples, respectively. PCR detected Trypanosoma congolense, subgenus Trypanozoon, Theileria equi, and Babesia caballi in 18 (9.1%), 77 (38.9%), 18 (9.1%), and 8 (4%) samples, respectively. Of the 77 Trypanozoon-positive samples, 35 (45.5%) were confirmed as Trypanosoma evansi type A. To our knowledge, this is the first report of detection of Trypanosoma congolense in donkeys outside of tsetse-infested areas in Sudan.
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http://dx.doi.org/10.1292/jvms.19-0534DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7118482PMC
March 2020

Prevalence of different trypanosomes in livestock in Blue Nile and West Kordofan States, Sudan.

Acta Trop 2020 Mar 16;203:105302. Epub 2019 Dec 16.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan; Research Center for Global Agromedicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan. Electronic address:

African animal trypanosomosis, transmitted cyclically by tsetse flies or mechanically by other biting flies, causes serious health problems in livestock. Although tsetse infestations have been observed in Blue Nile State in Sudan, tsetse was eradicated in West Kordofan in 1962, and no further studies have been carried out. Accordingly, in this study, we investigated the prevalence of trypanosomosis in cattle, sheep, and goats in Blue Nile and West Kordofan States, Sudan. This cross-sectional study was conducted using 70 cattle, 62 sheep, and 116 goats, and the microhematocrit centrifugation technique was used as a parasitological test. KIN-multispecies polymerase chain reaction (PCR) was used to detect Trypanozoon sp., Trypanosoma congolense, and T. vivax; RoTat 1.2 variable surface glycoprotein-specific PCR was used to detect T. evansi; and TviCatL PCR was used to specifically detect T. vivax. The seroprevalence of trypanosomosis was assessed using card agglutination tests CATT/ T. evansi. The parasitological prevalence rates were 4% (3/70) in cattle, 2% (1/62) in sheep, and 4% (5/116) in goats. The molecular prevalence rates of T. vivax, the most prevalent parasite, were 99% (69/70) in cattle, 98% (61/62) in sheep, and 84% (98/116) in goats. Trypanozoon (T. evansi or T. brucie) rates were 30% (21/70) in cattle, 32% (20/62) in sheep, and 12% (14/116) in goats. Among Trypanozoon-positive isolates, T. evansi was confirmed in 24% (5/21) of cattle, 70% (14/20) of sheep, and 86% (12/14) of goats. Finally, T. congolense was recorded only in cattle in Blue Nile State, with a prevalence of 14% (10/70). The seroprevalence rates of CATT/T. evansi were 46% (32/70) in cattle, 45% (28/62) in sheep, and 14% (16/116) in goats. Thus, we confirmed molecularly, for the first time, the presence of Trypanozoon, particularly T. evansi and T. vivax, in sheep and goats in Sudan. Our results show that sheep and goats could be an important reservoir for trypanosomes, potentially leading to the spread of the disease to the northern parts of the country following the movement of these animals. These findings provide important insights into the epidemiology of the disease and could affect the establishment of control strategies against trypanosomosis in Sudan.
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http://dx.doi.org/10.1016/j.actatropica.2019.105302DOI Listing
March 2020

A Seroepidemiological Survey of and in Horses in Mongolia.

J Parasitol 2019 08;105(4):580-586

2   National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.

Equine piroplasmosis caused by and is an economically important disease with a worldwide distribution. The objective of the present study was to investigate the seroepidemiology of and in horses reared in various Mongolian provinces. Serum samples prepared from blood collected from horses in 19 Mongolian provinces were screened for antibodies specific to and using enzyme-linked immunosorbent assays based on recombinant forms of merozoite antigen-2 and the 48-kDa merozoite rhoptry protein, respectively. Of 1,282 horses analyzed, 423 (33%) and 182 (14.2%) were sero-positive for and , respectively. Additionally, 518 (40.4%) were positive for at least 1 parasite species, of which 87 (16.8%) were co-infected with both parasites. Both and were detected in all surveyed provinces, and on a per province basis the positive rates ranged from 19.0 to 74.2% and 4.5 to 39.8%, respectively. - and -positive rates were comparable between male horses (31.9 and 14.1%, respectively) and female horses (34.5 and 14.3%, respectively). However, the positive rates were higher in the >3-yr-old age group (37.7 and 15.6%, respectively) compared with the 1-3-yr-old age group (19.4 and 10.0%, respectively). These findings confirmed that and infections are widespread among horses all over Mongolia, and that horse age is a risk factor for infection in this country. Our results will be useful for designing appropriate control measures to minimize and infections among Mongolian horses.
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August 2019

Detection of canine Schistosoma japonicum infection using recombinant thioredoxin peroxidase-1 and tandem repeat proteins.

J Vet Med Sci 2019 Oct 6;81(10):1413-1418. Epub 2019 Aug 6.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.

Humans and dogs live very close together and share various pathogens causing zoonotic parasitoses like schistosomiasis. A previous population genetics study done for schistosomes in the Philippines suggested that there is a high transmission level of Schistosoma japonicum among humans and dogs proving that the latter are important reservoirs for this zoonotic parasite. A more sensitive and specific test detecting schistosome infection in dogs will therefore strengthen the zoonotic surveillance, which might help in the possible elimination of this ancient disease. In this study, recombinant thioredoxin peroxidase-1 (SjTPx-1) and tandem repeat proteins (Sj1TR, Sj2TR, Sj4TR, Sj7TR) previously tested on human and water buffalo samples were used to assess its diagnostic applicability to dogs. Fifty-nine dog serum and stool samples were collected in the schistosomiasis-endemic municipalities of Calatrava, Negros Occidental and Catarman, Northern Samar in the Philippines and examined using the ELISA as compared to microscopy and fecal sample-based PCR. Samples positive for Babesia gibsoni and Dirofilaria immitis were also used to check for cross-reaction. Results showed that SjTPx-1 (80% sensitivity, 92.3% specificity) and Sj7TR (73.3% sensitivity, 92.3% specificity) have good potentials for diagnosing S. japonicum infection in dogs. These diagnostic antigens will therefore improve the surveillance in the transmission of the parasites from dogs to humans.
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http://dx.doi.org/10.1292/jvms.19-0126DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6863713PMC
October 2019

Genetic diversity and population structure of Glossina morsitans morsitans in the active foci of human African trypanosomiasis in Zambia and Malawi.

PLoS Negl Trop Dis 2019 07 25;13(7):e0007568. Epub 2019 Jul 25.

Division of Collaboration and Education, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Hokkaido, Japan.

The tsetse fly, Glossina morsitans morsitans, is a significant problem in Zambia and Malawi. It is the vector for the human infective parasite Trypanosoma brucei rhodesiense, which causes human African trypanosomiasis, and various Trypanosoma species, which cause African animal trypanosomiasis. Understanding the genetic diversity and population structure of G. m. morsitans is the basis of elucidating the connectivity of the tsetse fly populations, information that is essential in implementing successful tsetse fly control activities. This study conducted a population genetic study using partial mitochondrial cytochrome oxidase gene 1 (CO1) and 10 microsatellite loci to investigate the genetic diversity and population structure of G. m. morsitans captured in the major HAT foci in Zambia and Malawi. We have included 108 and 99 G. m. morsitans samples for CO1 and microsatellite analyses respectively. Our results suggest the presence of two different genetic clusters of G. m. morsitans, existing East and West of the escarpment of the Great Rift Valley. We have also revealed genetic similarity between the G. m. morsitans in Kasungu National Park and those in the Luangwa river basin in Zambia, indicating that this population should also be included in this historical tsetse belt. Although further investigation is necessary to illustrate the whole picture in East and Southern Africa, this study has extended our knowledge of the population structure of G. m. morsitans in Southern Africa.
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http://dx.doi.org/10.1371/journal.pntd.0007568DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6657825PMC
July 2019

Molecular detection and genetic characterisation of pathogenic Theileria, Anaplasma and Ehrlichia species among apparently healthy sheep in central and western Kenya.

Onderstepoort J Vet Res 2019 Jun 13;86(1):e1-e8. Epub 2019 Jun 13.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine.

Tick-borne diseases (TBDs) caused by Theileria, Babesia, Anaplasma and Ehrlichia species are common in tropical and subtropical regions. In this study, we investigated the presence and genetic diversity of Theileria spp., Anaplasma ovis, B. ovis, E. ruminantium and Anaplasma spp. in sheep from the Machakos and Homa Bay counties of Kenya. In order to improve the diagnosis and control of ovine TBDs, a total of 76 blood samples from apparently healthy sheep were screened using a polymerase chain reaction (PCR). The assays were conducted using primers based on Theileria spp. 18S rRNA, Anaplasma ovis Major surface protein-4 (AoMSP4), B. ovis 18S rRNA, E. ruminantium pCS20 and Anaplasma spp. 16S rRNA. The overall infection rates for Theileria spp., A. ovis, E. ruminantium and Anaplasma spp. were 39/76 (51.3%), 26/76 (34.2%), 6/76 (7.9%) and 31/76 (40.8%), respectively. The overall co-infection was 47/76 (61.8%). All Theileria spp. positive samples were confirmed to be of Theileria ovis on sequencing. A phylogenetic analysis of the 18S rRNA gene sequences of T. ovis revealed that all isolates of this study clustered with T. ovis sequences extracted from the GenBank suggesting this gene is highly conserved. E. ruminantium pCS20 sequences were in the same clade on the phylogenetic tree. However, three AoMSP4 sequences from this study appeared in the same clade, while one sequence formed a separate branch revealing genetic divergence. The 16S rRNA sequencing revealed uncharacterised Anaplasma spp. and A. ovis. The phylogenetic analyses of the uncharacterised Anaplasma spp. revealed that the two sequences from this study appear in an independent clade from other sequences extracted from the GenBank. This study provides important information regarding the occurrence of tick-borne pathogens and their degree of genetic diversity among sheep in Kenya, which is useful for the diagnosis and control of TBDs.
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http://dx.doi.org/10.4102/ojvr.v86i1.1630DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6620505PMC
June 2019

Polyradiculoneuropathy in dourine-affected horses.

Neuromuscul Disord 2019 06 18;29(6):437-443. Epub 2019 Mar 18.

Research Center for Global Agromedicine, Obihiro University of Agriculture and Veterinary Medicine, Nishi 2-11, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.

Dourine is an equine protozoan disease caused by Trypanosoma equiperdum. Dourine-afflicted animals die after developing neurological clinical signs, such as unilateral paresis. The disease has been a problem for many years; however, the pathogenesis regarding the neurological clinical signs of dourine has been unclear. In the present study, we conducted a histopathological examination in order to investigate the mechanisms by which dourine-afflicted horses develop the accompanying neurological clinical signs. Four dourine-afflicted horses in Mongolia were evaluated. An apparently healthy horse exhibited multifocal neuritis without axonal or myelin degeneration. The other horses, which had obvious neurological clinical signs, also exhibited multifocal neuritis. In particular, the nerves that innervated areas associated with neurological clinical signs exhibited neuritis with demyelination in the latter horses. Inflamed, non-demyelinating nerves were infiltrated with B lymphocytes and T lymphocytes; while inflamed, demyelinating nerves were infiltrated with mononuclear phagocytes. Our observations revealed lesion progression in the nerves, such that polyradiculoneuropathy could explain the accompanying neurological clinical signs of dourine. To our knowledge, this is the first report to describe a pathogenic mechanism for the development of the neurological clinical signs found in dourine-afflicted horses.
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http://dx.doi.org/10.1016/j.nmd.2019.03.005DOI Listing
June 2019

Equine trypanosomosis: enigmas and diagnostic challenges.

Parasit Vectors 2019 May 15;12(1):234. Epub 2019 May 15.

Department of Biomedical Sciences, Institute of Tropical Medicine, Nationalestraat 155, 2000, Antwerp, Belgium.

Equine trypanosomosis is a complex of infectious diseases called dourine, nagana and surra. It is caused by several species of the genus Trypanosoma that are transmitted cyclically by tsetse flies, mechanically by other haematophagous flies, or sexually. Trypanosoma congolense (subgenus Nannomonas) and T. vivax (subgenus Dutonella) are genetically and morphologically distinct from T. brucei, T. equiperdum and T. evansi (subgenus Trypanozoon). It remains controversial whether the three latter taxa should be considered distinct species. Recent outbreaks of surra and dourine in Europe illustrate the risk and consequences of importation of equine trypanosomosis with infected animals into non-endemic countries. Knowledge on the epidemiological situation is fragmentary since many endemic countries do not report the diseases to the World Organisation for Animal Health, OIE. Other major obstacles to the control of equine trypanosomosis are the lack of vaccines, the inability of drugs to cure the neurological stage of the disease, the inconsistent case definition and the limitations of current diagnostics. Especially in view of the ever-increasing movement of horses around the globe, there is not only the obvious need for reliable curative and prophylactic drugs but also for accurate diagnostic tests and algorithms. Unfortunately, clinical signs are not pathognomonic, parasitological tests are not sufficiently sensitive, serological tests miss sensitivity or specificity, and molecular tests cannot distinguish the taxa within the Trypanozoon subgenus. To address the limitations of the current diagnostics for equine trypanosomosis, we recommend studies into improved molecular and serological tests with the highest possible sensitivity and specificity. We realise that this is an ambitious goal, but it is dictated by needs at the point of care. However, depending on available treatment options, it may not always be necessary to identify which trypanosome taxon is responsible for a given infection.
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http://dx.doi.org/10.1186/s13071-019-3484-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6518633PMC
May 2019

Molecular characterization of a new Trypanosoma (Megatrypanum) theileri isolate supports the two main phylogenetic lineages of this species in Japanese cattle.

Parasitol Res 2019 Jun 4;118(6):1927-1935. Epub 2019 May 4.

Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, Japan.

Trypanosoma (Megatrypanum) theileri is a cosmopolitan, usually non-pathogenic, trypanosome of cattle transmitted by blood-sucking arthropods, mainly tabanid flies. Several T. theileri strains isolated from domestic and wild ruminants via co-culturing with mammalian feeder cells or blood cells have been characterized morphologically and genetically. Here, we cultured a new trypanosome isolate from a Holstein cow in Hokkaido, Japan, and performed morphological and molecular characterization studies. The new isolate (Obihiro strain) was co-cultivated with Madin-Darby bovine kidney (MDBK) cells in GIT medium supplemented with 10% fetal bovine serum. Trypomastigotes and epimastigotes, but not intracellular parasites, were identified in the culture. Analysis of the V7-V8 region of 18S rRNA sequences showed that the Obihiro strain is positioned within the subgenus Megatrypanum. A dendrogram based on whole internal transcribed spacer rDNA sequence showed that the Obihiro strain clustered in the lineage TthII together with the Japanese isolates of T. theileri, Esashi 9, and Esashi 12, and isolates from Zambia and the USA. T. theileri of the KM strain and a T. theileri-like trypanosome isolated from deer (TSD1 strain) clustered in the lineage TthI, separate from the Obihiro strain. Based on a partial cathepsin L-like protein gene analysis, the Obihiro strain clustered with isolates of the TthIIF genotype, which includes T. theileri from Vietnam, Sri Lanka, and Brazil. Our analyses of the T. theileri Obihiro strain provide relevant insights into its genetic diversity in Japanese cattle and corroborate the host specificity of cattle and deer trypanosomes of the subgenus Megatrypanum.
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http://dx.doi.org/10.1007/s00436-019-06313-xDOI Listing
June 2019

Utilization of crude and recombinant ELISAs for serodiagnosis of camel trypanosomosis in Sudan.

Vet Parasitol Reg Stud Reports 2019 04 27;16:100278. Epub 2019 Feb 27.

Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.

This study was carried out to evaluate the application of CATT/T. evansi, crude and recombinant (TeGM6-4r) antigen ELISAs in the diagnosis of camel trypanosomosis caused by two trypanosome species, T. evansi and T. vivax, in Sudan. Concurrently, the current situation of camel trypanosomosis was investigated based on the results of a serological analysis. The recombinant tandem repeat antigen TeGM6-4r is conserved among salivarian trypanosome species and was highly sensitive in the detection Trypanozoon, and T. vivax. It has been validated in the diagnosis of surra in cattle and water buffalo but not in camels. A comparative evaluation of a crude antigen ELISA and a recombinant antigen GM6 (rTeGM6-4r) ELISA was performed using 189 blood samples, which included 148 samples obtained from different camel herds in Eastern Sudan and 41 samples from camels that had been brought from Western Sudan to local markets. The results showed that the rTeGM6-4r ELISA detected the greatest number of positive samples (n = 118, 62%), while CATT/T. evansi and the crude antigen ELISA detected the lowest number of positive samples (n = 73, 39%). The kappa value of rTeGM6-4r as compared to TeCA ELISA was 0.5515, which indicated moderate agreement. We concluded that the rTeGM6-4r ELISA is the test of choice for use in screening camel for trypanosomosis caused by T. evansi and T. vivax in Sudan.
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http://dx.doi.org/10.1016/j.vprsr.2019.100278DOI Listing
April 2019

Identification and genetic characterization of Piroplasmida and Anaplasmataceae agents in feeding Amblyomma variegatum ticks from Benin.

Vet Parasitol Reg Stud Reports 2018 12 29;14:137-143. Epub 2018 Oct 29.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-855, Japan; Research Center for Global Agromedicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-855, Japan. Electronic address:

In many African countries including Benin, the reluctance of some livestock owners to blood collection from their cattle makes epidemiological surveys cumbersome and prevents regular monitoring of tick-borne diseases. In the present study, Amblyomma variegatum ticks were used to find out more about bovine tick-borne pathogens. DNA extracts from 910 adult ticks collected off cattle in North East Benin were examined for Babesia bigemina, B. bovis, Theileria taurotragi, T. annulata, T. orientalis, T. parva, T. mutans, Anaplasma marginale and Ehrlichia ruminantium using pathogen-specific PCR assays and sequence analyses. Altogether, 21.6% of the ticks carried at least one pathogen. A. marginale (142/910) was the most frequent pathogen, followed by E. ruminantium (57/910), B. bovis (10/910), T. mutans (3/910) and B. bigemina (1/910). Theileria taurotragi, T. annulata, T. orientalis, T. parva were not detected in the samples. Babesia bigemina, B. bovis and T. mutans were present in only one location whereas A. marginale and E. ruminantium were found in ticks from 7/8 locations surveyed. Coinfections occurred in 7.1% of all positive ticks. The analyses of partial sequences of B. bovis spherical body protein 4, B. bigemina rhoptry-associated protein-1a, T. mutans 18S rRNA and A. marginale major surface protein 5 showed high sequence conservation and homologies between Benin isolates and those from other African countries. However, E. ruminantium pCS20 partial sequences were different from published West African isolates and presented similar genetic variation with South and East African isolates. These results provide information on the pathogens circulating in North East Benin and suggest that Am. variegatum, one of the most abundant ticks in Africa, may play a role in the transmission of A. marginale.
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http://dx.doi.org/10.1016/j.vprsr.2018.10.006DOI Listing
December 2018

Genetic characterization of tick-borne pathogens in ticks infesting cattle and sheep from three South African provinces.

Ticks Tick Borne Dis 2019 06 15;10(4):875-882. Epub 2019 Apr 15.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, 080-8555, Japan. Electronic address:

Ticks are involved in the transmission of many public health and veterinary important pathogens. Although tick-borne pathogens are widely distributed in South Africa, information on tick-pathogen relationship needs to be updated particularly using modern molecular techniques. This study used PCR and sequencing to confirm the identity of the tick species collected from cattle and sheep from KwaZulu-Natal, Free State and Eastern Cape. Furthermore, presence of Babesia spp., Theileria spp., Anaplasma marginale, Rickettsia spp., Ehrlichia ruminantium and Coxiella burnetii was detected from tick DNA using species-specific PCR or nested PCRs. The study samples consisted of 390 adult ticks (male and female) which were pooled according to species, host animal and sampling site (three ticks per pool) for DNA extraction. The PCR results revealed that out of 130 tick DNA pools, 30 (23.1%) were positive for at least one pathogen. The most frequent pathogen was C. burnetii (9.2%), followed by Rickettsia spp. (7.7%), A. marginale (3.8%), T. mutans (3.1%), T. taurotragi (2.3%) and E. ruminantium (1.5%). The highest prevalence of pathogens was observed in ticks collected from cattle in Eastern Cape (16/42) and the lowest was in ticks obtained from sheep in Free State (1/21). Infected ticks were identified as Rhipicephalus evertsi evertsi (n = 13), R. appendiculatus (n = 3), R. decoloratus (n = 7) and Amblyomma hebraeum (n = 7). Coinfection with two pathogens was found in 21% of pathogen-positive pools. Analysis of Theileria taurotragi 18S rRNA, T. mutans 18S rRNA, C. burnetii htpB, Rickettsia spp. gltA, Rickettsia spp. ompA, E. ruminantium pCS20 and A. marginale Msp5 sequences showed that the pathogens detected in this study were genetically related to isolates previously reported in Africa. These findings provide important information on distribution of ticks and tick-borne pathogens of ruminants and will contribute in the formulation of future control strategies in South Africa.
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http://dx.doi.org/10.1016/j.ttbdis.2019.04.008DOI Listing
June 2019

Short- and long-term effects of orally administered azithromycin on Trypanosoma brucei brucei-infected mice.

Exp Parasitol 2019 Apr 3;199:40-46. Epub 2019 Mar 3.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Nishi 2-11 Inada, Obihiro, Hokkaido, 080-8555, Japan; Research Center for Global Agromedicine, Obihiro University of Agriculture and Veterinary Medicine, Nishi 2-11 Inada, Obihiro, Hokkaido, 080-8555, Japan. Electronic address:

Human African trypanosomosis (HAT) and animal African trypanosomosis (AAT) are diseases of economic importance in humans and animals that affect more than 36 African countries. The currently available trypanocidal drugs are associated with side effects, and the parasites are continually developing resistance. Thus, effective and safe drugs are needed for the treatment of HAT and AAT. This study aimed to evaluate the effects of azithromycin (AZM) on Trypanosoma brucei brucei-infected mice. Mice were randomly divided into 7 groups consisting of a vehicle control group, 5 test groups and a diminazene aceturate (DA)-treated group. Mice were treated orally for 7 and 28 days, as short-term and long-term treatments, respectively. Short-term AZM treatment cured 23% (16 of 70) of the overall treated mice whereas long-term treatment resulted in the survival of 70% of the mice in the groups that received AZM at doses of 300 and 400 mg/kg. Trypanosomes treated in vitro with 25 μg/mL of AZM were subjected to transmission electron microscopy, which revealed the presence of increased numbers of glycosomes and acidocalcisomes in comparison to the vehicle group. The current study showed the trypanocidal effect of AZM on T. b. brucei in vivo. The demonstrated efficacy increased with an increase in treatment period and an increased concentration of AZM.
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http://dx.doi.org/10.1016/j.exppara.2019.02.018DOI Listing
April 2019

Draft Genome Sequence of Strain IVM-t1.

Microbiol Resour Announc 2019 Feb 28;8(9). Epub 2019 Feb 28.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan.

Trypanosoma equiperdum primarily parasitizes the genital organs and causes dourine in equidae. We isolated a new T. equiperdum strain, T. equiperdum IVM-t1, from the urogenital tract of a horse definitively diagnosed as having dourine in Mongolia. Here, we report the whole-genome sequence, the predicted gene models, and their annotations.
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http://dx.doi.org/10.1128/MRA.01119-18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6395869PMC
February 2019

Molecular detection of Anaplasma ovis in small ruminants and ixodid ticks from Mongolia.

Parasitol Int 2019 Apr 17;69:47-53. Epub 2018 Nov 17.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan. Electronic address:

Anaplasma ovis is a tick-borne obligate intracellular rickettsial bacterium that causes anaplasmosis in domestic and wild small ruminants. Sheep and goats, whose combined population is approximately 48.5-million in Mongolia, play a vital role in the country's economy. In this study, we conducted an epidemiological survey of A. ovis in sheep and goats from 19 of 21 provinces in Mongolia. Additionally, DNA samples extracted from unfed ticks collected in 11 Mongolian provinces were also screened for A. ovis. Of 1179 and 871 blood DNA samples from sheep and goats, 813 (69.0%) and 621 (71.3%), respectively, were positive for A. ovis when screened by a PCR assay based on major surface protein 4 gene (msp4). On a per province basis, A. ovis infection rates ranged from 7.4%-93.3% and 13.3%-100% in sheep and goats, respectively. Subsequently, DNA samples prepared from 721 unfed ticks, including Dermacentor nuttalli (n = 378), Ixodes persulcatus (n = 95), Haemaphysalis pospelovashtromae (n = 120), and Hyalomma asiaticum (n = 128), were screened for A. ovis using the same PCR assay. Although nine D. nuttalli were A. ovis-positive, all other tick DNA samples were negative. In addition to reporting A. ovis in sheep and goats from all over Mongolia, this study identified D. nuttalli as a potential transmission vector of A. ovis in Mongolia. The present data highlight the importance of monitoring Mongolian sheep and goats for possible episodes of clinical anaplasmosis and controlling D. nuttalli throughout the country.
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http://dx.doi.org/10.1016/j.parint.2018.11.004DOI Listing
April 2019

Molecular detection and characterization of tick-borne protozoan and rickettsial pathogens isolated from cattle on Pemba Island, Tanzania.

Ticks Tick Borne Dis 2018 09 4;9(6):1437-1445. Epub 2018 Jul 4.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, 080-8555, Japan. Electronic address:

Tick-borne diseases cause significant losses to livestock production in tropical and subtropical regions. In Tanzania, detailed studies on tick-borne pathogens in cattle using sensitive molecular detection methods are scarce. In this study, we investigated the occurrence of Theileria spp., Babesia spp., Anaplasma spp. and Ehrlichia spp. in 245 blood samples collected from cattle on Pemba Island, Tanzania. We used polymerase chain reaction (PCR) and gene sequencing to detect and identify pathogens. PCR screening revealed overall infection rates of 62.4% for Theileria spp., 17.6% for Babesia bigemina, 15.9% for Anaplasma marginale, 7.4% for Ehrlichia ruminantium and 4.5% for Babesia bovis. Further analysis using sequences of Theileria spp. 18S rRNA revealed infection of cattle with Theileria mutans (68.6%), Theileria taurotragi (48.4%), Theileria parva (41.2%), and Theileria ovis (1.9%). Co-infections of cattle, with up to six tick-borne pathogens, were revealed in 46.9% of the samples. Sequence analysis indicated that T. parva p104, E. ruminantium pCS20 and A. marginale MSP-5 genes are conserved among cattle blood samples in Pemba, with 99.3%-100%, 99.6%-100% and 100% sequence identity values, respectively. In contrast, the B. bigemina RAP-1a and B. bovis SBP-2 gene sequences were relatively diverse with 99.5%-99.9% and 66.4%-98.7% sequence identity values respectively. The phylogenetic analyses revealed that T. parva p104, E. ruminantium pCS20 and A. marginale MSP-5 gene sequences clustered in the same clade with other isolates from other countries. In contrast, the B. bigemina RAP-1 and B. bovis SBP-2 gene sequences showed significant differences in the genotypes, as they appeared in separate clades. This study provides important data for understanding the epidemiology of tick-borne diseases, and is expected to improve the approach for diagnosis and control of tick-borne diseases in Tanzania.
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http://dx.doi.org/10.1016/j.ttbdis.2018.06.014DOI Listing
September 2018

The utility of an rTeGM6-4r-based immunochromatographic test for the serological diagnosis of non-tsetse-transmitted equine trypanosomosis in rural areas of Mongolia.

Parasitol Res 2018 Sep 25;117(9):2913-2919. Epub 2018 Jun 25.

National Research Center for Protozoan Diseases, OIE Reference Laboratory for Surra, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Hokkaido, 080-8555, Japan.

Our previous studies report epidemics of non-tsetse-transmitted equine trypanosomosis in Mongolia. However, the current status of non-tsetse-transmitted equine trypanosomosis endemicity remains to be clarified in some parts of Mongolia. We previously reported the potential application of rTeGM6-4r-based diagnostic tools, an rTeGM6-4r-based immunochromatographic test (ICT) and an enzyme-linked immunosorbent assay (ELISA), in the serological surveillance of equine trypanosomosis in Mongolia. In the present study, the utility of the rTeGM6-4r-based ICT was validated. The rTeGM6-4r-based ICT accurately diagnosed positive reference sera that had been prepared from dourine horses in Mongolia, similarly to the rTeGM6-4r-based ELISA. The diagnostic performance of the rTeGM6-4r-based ICT was maintained when the strips were preserved for at least 2 months under dry conditions. The ICT detected 42 positive serum samples from a total of 1701 equine sera that had been collected from all 21 provinces of Mongolia. The κ-value, sensitivity and specificity of rTeGM6-4r-based ICT were 0.58, 50.0% (95% CI, 37.7-62.3%) and 99.3% (95% CI, 98.7-99.6%), respectively, in comparison to the rTeGM6-4r-based ELISA. Our field-friendly rTeGM6-4r-based ICT was found to be useful for the serological diagnosis of non-tsetse-transmitted equine trypanosomosis in rural areas of Mongolia.
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http://dx.doi.org/10.1007/s00436-018-5982-8DOI Listing
September 2018

Transovarial persistence of Babesia ovata DNA in a hard tick, Haemaphysalis longicornis, in a semi-artificial mouse skin membrane feeding system.

Acta Parasitol 2018 06;63(2):433

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-Cho, Obihiro, Hokkaido 080-8555, Japan.

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http://dx.doi.org/10.1515/ap-2018-0051DOI Listing
June 2018

Detection and molecular characterization of tick-borne pathogens infecting sheep and goats in Blue Nile and West Kordofan states in Sudan.

Ticks Tick Borne Dis 2018 03 1;9(3):598-604. Epub 2018 Feb 1.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan; Research Center for Global Agromedicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan. Electronic address:

Tick-borne pathogens (TBPs) are common in livestock of sub-Saharan Africa. However, information regarding TBPs in sheep and goats in Sudan is limited. In this study, 178 blood samples of sheep and goats in Blue Nile and West Kordofan states were investigated for TBPs using PCR. Overall, 110 (61.8%) samples were found to be infected with at least one of the following pathogens: Anaplasma ovis, Theileria ovis, and Ehrlichia ruminantium. Babesia ovis and T. lestoquardi were not identified. A. ovis was the most prevalent pathogen (n = 107, 60.1%), followed by T. ovis (n = 23, 12.9%) and E. ruminantium (n = 1, 0.6%). The prevalence rates of A. ovis and T. ovis were significantly higher in sheep than in goats. Phylogenetic analysis of T. ovis 18S rRNA and A. ovis msp4, groEL, and 16S rRNA, revealed that the pathogens identified in this study are clustered together, indicating similar molecular characteristics. Additionally, phylogenetic analysis of E. ruminantium pCS20 revealed that E. ruminantium in this study belong to the West Africa group, and different to E. ruminantium previously identified in ticks from Sudan. We concluded that TBPs are highly prevalent in the study area and continuous monitoring of TBPs in sheep and goats in Sudan is highly required.
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http://dx.doi.org/10.1016/j.ttbdis.2018.01.014DOI Listing
March 2018

Evaluation of Schistosoma japonicum thioredoxin peroxidase-1 as a potential circulating antigen target for the diagnosis of Asian schistosomiasis.

J Vet Med Sci 2018 Feb 28;80(1):156-163. Epub 2017 Nov 28.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.

Asian schistosomiasis caused by Schistosoma japonicum is a serious zoonotic disease endemic in China, the Philippines and parts of Indonesia. Mass drug administration in endemic areas resulted to decline in disease severity and intensity. The low intensity of infection limits the use of current parasitological methods for schistosomiasis diagnosis. Detection of parasite circulating antigens might provide more informative result as it may indicate the true status of infection. In this study, S. japonicum thioredoxin peroxidase-1 (SjTPx-1) a 22 kDa secreted antioxidant enzyme expressed throughout the life stages of the parasite was evaluated for its potential use as a biomarker for schistosomiasis japonica infection. Rabbit polyclonal antibody and mouse monoclonal antibodies (mAbs) were raised against the recombinant SjTPx-1 (rSjTPx-1). The antibodies produced against the recombinant antigen was confirmed to detect the native SjTPx-1 in crude adult worm lysate. Likewise, the specific binding of mAbs to parasite TPx-1 and not to mammalian peroxiredoxin-1 orthologues was also confirmed. The double antibody sandwich ELISA developed in this study was able to detect at least 1 ng/ml of rSjTPx-1. In addition, this method was able to detect the antigen from all serum samples of experimentally infected rabbit and mice. The diagnostic potential of SjTPx-1 in human clinical samples was also evaluated, in which 4 out of 10 stool-confirmed serum samples had detectable levels of the antigen. The results suggest that SjTPx-1 can be a potential biomarker for Asian zoonotic schistosomiasis.
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http://dx.doi.org/10.1292/jvms.17-0579DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5797876PMC
February 2018

Identification and characterization of interchangeable cross-species functional promoters between Babesia gibsoni and Babesia bovis.

Ticks Tick Borne Dis 2018 02 21;9(2):330-333. Epub 2017 Nov 21.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido, 080-8555, Japan. Electronic address:

The development of transgenic techniques has been reported in many protozoan parasites over the past few years. We recently established a successful transient transfection system for Babesia gibsoni based on Bg 5'-ef-1α promoter. This study investigated 6 homologous and 6 heterologous promoters for B. gibsoni and B. bovis and identified novel interchangeable cross-species functional promoters between B. gibsoni and B. bovis. Ten out of twelve promoters had heterologous promoter function. In particular, Bg 5'-ef-1α and Bg 5'-actin heterologous promoters resulted in a significantly higher luciferase activity than Bb 5'-ef-1α homologous promoter in B. bovis. The present study showed that Bg 5'-actin promoted the highest luciferase activity in both B. gibsoni and B. bovis. The study further indicates that heterologous promoter function widely exists between B. gibsoni and B. bovis. This finding is an important step for future stable transfection construct design and for the production of vaccines based on transfected B. gibsoni and B. bovis parasites.
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http://dx.doi.org/10.1016/j.ttbdis.2017.11.008DOI Listing
February 2018

Molecular analysis of tick-borne protozoan and rickettsial pathogens in small ruminants from two South African provinces.

Parasitol Int 2018 Apr 15;67(2):144-149. Epub 2017 Nov 15.

National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan. Electronic address:

Tick-borne protozoan and rickettsial diseases are a major threat to livestock in tropical and sub-tropical regions of Africa. In this study we investigated the presence and distribution of Theileria spp., Babesia ovis, Anaplasma ovis, Anaplasma phagocytophilum, Ehrlichia ruminantium and SFG Rickettsia in sheep and goats from Free State and KwaZulu-Natal provinces. A total of 91 blood samples were screened in this study, 61 from goats and 30 from sheep. PCR assay was conducted using primers based on Theileria spp. 18S rRNA, Babesia ovis (BoSSU rRNA), Anaplasma ovis (AoMSP4), Anaplasma phagocytophilum epank1, Ehrlichia ruminantium pCS20 and SFG Rickettsia OmpA. Overall infection rates of Theileria spp., Anaplasma ovis and Ehrlichia ruminantium were 18 (19.8%), 33 (36.3%) and 13 (14.3%), respectively. The co-infection of two pathogens were detected in 17/91 (18.7%) of all samples, goats having higher rates of co-infection compared to sheep. Phylogenetic tree analysis sequence of pCS20 gene of E. ruminantium of this study was found to be in the same clade with Kumm2 and Riverside strains both from South Africa. The phylogram of SSU rRNA of Theileria ovis had longer branch length compared to all other sequences most of which were from Asia and Middle East. This study provides important data for understanding the tick-borne diseases occurrence in the study area and it is expected to improve the approach for the diagnosis and control of these diseases.
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http://dx.doi.org/10.1016/j.parint.2017.11.002DOI Listing
April 2018

Transovarial persistence of Babesia ovata DNA in a hard tick, Haemaphysalis longicornis, in a semi-artificial mouse skin membrane feeding system.

Acta Parasitol 2017 Dec;62(4):836-841

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Bovine piroplasmosis, a tick-borne protozoan disease, is a major concern for the cattle industry worldwide due to its negative effects on livestock productivity. Toward the development of novel therapeutic and vaccine approaches, tick-parasite experimental models have been established to clarify the development of parasites in the ticks and the transmission of the parasites by ticks. A novel tick-Babesia experimental infection model recently revealed the time course of Babesia ovata migration in its vector Haemaphysalis longicornis, which is a dominant tick species in Japan. However, there has been no research on the transovarial persistence of B. ovata DNA using this experimental infection model. Here we assessed the presence of B. ovata DNA in eggs derived from parthenogenetic H. longicornis female ticks that had engorged after semi-artificial mouse skin membrane feeding of B. ovata-infected bovine red blood cells. The oviposition period of the engorged female ticks was 21-24 days in the semi-artificial feeding. Total egg weight measured daily reached a peak by day 3 in all female ticks. Nested PCR revealed that 3 of 10 female ticks laid B. ovata DNA-positive eggs after the semi-artificial feeding. In addition, B. ovata DNA was detected at the peak of egg weight during oviposition, indicating that B. ovata exist in the eggs laid a few days after the onset of oviposition in the tick. These findings will contribute to the establishment of B. ovata-infected H. longicornis colonies under laboratory conditions.
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http://dx.doi.org/10.1515/ap-2017-0100DOI Listing
December 2017
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