Papers - HIRAI Takuya
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Detection of bovine respiratory disease complex-related pathogens in nasopharynx-associated lymphoid tissue. Reviewed
Hegazy AA, Nakai M, Fuke N, Hussein AE, Kondo H, Hirai T.
J Vet Diagn Invest 37 ( 2 ) 284 - 297 2025.3
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Histopathological findings of the nasopharynx-associated lymphoid tissue of pigs co-infected with porcine circovirus 2 and porcine reproductive and respiratory syndrome virus. Reviewed
Toyama K, Hirai T, Sueyoshi M, Zhou Z, Niazi AM, Kawaguchi N, Fuke N, Yamaguchi R.
J Vet Med Sci. 2022.10
Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)
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Niazi A.M., ZiHeng Z., Fuke N., Toyama K., Habibi W.A., Kawaguchi N., Yamaguchi R., Hirai T.
Journal of Comparative Pathology 197 23 - 34 2022.9
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Journal of Comparative Pathology
Porcine respiratory disease complex, which is caused by a combination of pathogens, including swine influenza A virus (SIV) and porcine reproductive respiratory syndrome virus (PRRSV), results in significant economic losses in pig production systems. Nasopharynx-associated lymphoid tissue (NALT) plays an important role in the uptake of pathogens and defence of the nasal mucosa in rodents and humans. We characterized NALT M cells in pigs and detected SIV antigen and PRRSV nucleic acid in NALT using histopathological, immunohistochemical and in-situ hybridization analyses. All SIV- and PRRSV-positive cases examined had suppurative nasopharyngitis and pneumonia. M cells were detected by immunohistochemistry and the distribution of M cells showed an increase in the middle section of NALT. SIV antigen was detected in M cells and PRRSV nucleic acid was demonstrated in the cytoplasm of macrophages in NALT. We believe that SIV and PRRSV infection in the upper respiratory tract induces local immunosuppression and these results confirm that swine NALT is a location for virus replication and may be strongly associated with the development of pneumonia in pigs.
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Rhabdomyolysis, myoglobinuric nephrosis, and crystalline nephropathy in a captive bottlenose dolphin Reviewed
Nueangphuet P., Hamano T., Hirai T., Sakaguchi Y., Sonoda H., Otsuka M., Yamato O., Hobo S., Ikeda M., Yamaguchi R.
Journal of Veterinary Diagnostic Investigation 34 ( 4 ) 668 - 673 2022.7
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Journal of Veterinary Diagnostic Investigation
A 5-y-old female bottlenose dolphin (Tursiops truncatus) from an aquarium in Japan had clinical signs of anorexia, vomiting, and bradykinesia. Enrofloxacin and lactated Ringer solution were administered for treatment of bacterial infection and for rehydration. Elevations of creatine kinase and aspartate aminotransferase activities were detected on day 4 of treatment, indicating that rhabdomyolysis had developed on day 3. On day 5, serum creatinine and urea concentrations increased and remained high throughout the remaining treatment; the dolphin died on day 16. Postmortem examination revealed massive necrosis of the longissimus dorsi muscles. Histologic examination revealed extensive necrosis of skeletal myofibers, multifocal renal tubular necrosis with intratubular casts and crystals, and suppurative bronchopneumonia. The renal casts labeled positively with anti-myoglobin antibody; expression of aquaporin-1 was decreased in renal tubules compared to normal kidney tissue. To our knowledge, this description of clinicopathologic findings of rhabdomyolysis leading to acute kidney injury with concomitant crystalline nephropathy has not been reported previously in a bottlenose dolphin.
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Niazmand M., Hirai T., Ito S., Habibi W., Noori J., Hasheme R., Yamaguchi R.
Journal of Wildlife Diseases 55 ( 4 ) 804 - 811 2019.10
Language：English Publishing type：Research paper (scientific journal) Publisher：Journal of Wildlife Diseases
We performed postmortem examinations on seven Misaki feral horses (Equus caballus) and evaluated Misaki feral horses, Japanese wild boars (Sus scrofa leucomystax), domestic pigs (Sus scrofa), and wild Japanese macaques (Macaca fuscata fuscata) from 2015 to 2017 in Cape Toi, Kushima, Miyazaki Prefecture, southern Japan, for antibodies against Japanese encephalitis virus (JEV). Strongylus vulgaris infection with severe arterial lesions and hemomelasma ilei was present in all necropsied horses. We frequently found intestinal ulcers, perihepatitis filamentosa, and poor body condition. We recorded degenerative arthropathy in metacarpophalangeal joints in two cases and a fracture of the rib with diaphragmatic rupture in one case. A total of 73% (177/242) of horses were seropositive for JEV as tested by hemagglutination inhibition (HI). The HI data also revealed that 74% (59/80) of the wild boars, 67% (60/90) of the pigs, and 29% (22/75) of the wild monkeys were seropositive for JEV. Our findings showed that Strongylus spp. are still a risk to horses in this region, and that environmental factors such as topographic location of the pasture and steep slope may have caused of degenerative arthropathy and bone fracture. Our results showed that JEV is endemic in Japan. The wild boars and pigs were presumed to act as strong amplifiers and sources of infection, with subsequent risk to humans.
DOI： 10.7589/2018-10-265
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Experimental induction of chicken amyloid A amyloidosis in white layer chickens by inoculation with inactivated vaccines.
Habibi WA, Hirai T, Niazmand MH, Okumura N, Yamaguchi R
Avian pathology : journal of the W.V.P.A 46 ( 5 ) 1 - 9 2017.5
Language：Japanese Publishing type：Research paper (scientific journal)
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Enhanced detection of Porcine reproductive and respiratory syndrome virus in fixed tissues by in situ hybridization following tyramide signal amplification.
Trang NT, Hirai T, Ngan PH, Lan NT, Fuke N, Toyama K, Yamamoto T, Yamaguchi R
Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 27 ( 3 ) 326 - 31 2015.5
Language：Japanese Publishing type：Research paper (scientific journal)
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Membranoproliferative glomerulonephritis in a calf with nephrotic syndrome. Reviewed
Trang NT, Hirai T, Nabeta R, Fuke N, Yamaguchi R
Journal of comparative pathology 151 ( 2-3 ) 162 - 5 2014.8
Language：English Publishing type：Research paper (scientific journal) Publisher：Journal of Comparative Pathology
A 2-month-old Japanese black calf was presented with a history of weight loss, exophthalmos and subcutaneous oedema of the brisket. Urinalysis and serum biochemistry showed proteinuria and hypoproteinaemia suggestive of nephrotic syndrome. Microscopically, lesions in the kidney were characterized by proliferation of mesangial cells and diffuse thickening of the glomerular basement membranes with the appearance of double contours. Immune complex deposits were confirmed by electron microscopy and immunofluorescence using reagents specific for bovine immunoglobulin G, complement factor C3 and bovine viral diarrhoea virus (BVDV). Consequently, the glomerular lesion in this case was diagnosed as membranoproliferative glomerulonephritis. BVDV type 1 was detected in serum by nested reverse transcriptase polymerase chain reaction. Viral antigen was also identified in the glomeruli by immunofluorescence. These results suggest that BVDV may have been the cause of immune complex glomerulonephritis in this calf. © 2014 Elsevier Ltd.
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Detection of porcine reproductive and respiratory syndrome virus in oral fluid from naturally infected pigs in a breeding herd.
Trang NT, Hirai T, Yamamoto T, Matsuda M, Okumura N, Giang NT, Lan NT, Yamaguchi R
Journal of veterinary science 15 ( 3 ) 361 - 7 2014
Language：Japanese Publishing type：Research paper (scientific journal)
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Mekata H., Yamada K., Umeki K., Yamamoto M., Ochi A., Umekita K., Kobayashi I., Hirai T., Okabayashi T.
BMC Veterinary Research 20 ( 1 ) 2024.12
Publishing type：Research paper (scientific journal) Publisher：BMC Veterinary Research
Severe fever with thrombocytopenia syndrome (SFTS) is a fatal zoonosis caused by ticks in East Asia. As SFTS virus (SFTSV) is maintained between wildlife and ticks, seroepidemiological studies in wildlife are important to understand the behavior of SFTSV in the environment. Miyazaki Prefecture, Japan, is an SFTS-endemic area, and approximately 100 feral horses, called Misaki horses (Equus caballus), inhabit Cape Toi in Miyazaki Prefecture. While these animals are managed in a wild-like manner, their ages are ascertainable due to individual identification. In the present study, we conducted a seroepidemiological survey of SFTSV in Misaki horses between 2015 and 2023. This study aimed to understand SFTSV infection in horses and its transmission to wildlife. A total of 707 samples from 180 feral horses were used to determine the seroprevalence of SFTSV using enzyme-linked immunosorbent assay (ELISA). Neutralization testing was performed on 118 samples. In addition, SFTS viral RNA was detected in ticks from Cape Toi and feral horses. The overall seroprevalence between 2015 and 2023 was 78.5% (555/707). The lowest seroprevalence was 55% (44/80) in 2016 and the highest was 92% (76/83) in 2018. Seroprevalence was significantly affected by age, with 11% (8/71) in those less than one year of age and 96.7% (435/450) in those four years of age and older (p < 0.0001). The concordance between ELISA and neutralization test results was 88.9% (105/118). SFTS viral RNA was not detected in ticks (n = 516) or feral horses. This study demonstrated that horses can be infected with SFTSV and that age is a significant factor in seroprevalence in wildlife. This study provides insights into SFTSV infection not only in horses but also in wildlife in SFTS-endemic areas.
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Hepatocellular carcinoma with lung metastasis showing hemochromatosis in an Egyptian fruit bat (Rousettus aegyptiacus). Reviewed
Kawaguchi N, Fuke N, Nueangphuet P, Pornthummawat A, Niazi AM, Izzati UZ, Hirai T, Yamaguchi R.
J Vet Med Sci. 10 ( 86 ) 49 - 53 2024.1
Language：English Publishing type：Research paper (scientific journal)
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Tanaka Y.L., Shofa M., Butlertanaka E.P., Niazi A.M., Hirai T., Mekata H., Saito A.
Pathogens 13 ( 1 ) 2024.1
Publishing type：Research paper (scientific journal) Publisher：Pathogens
Pigs are important animals for meat production but can carry several zoonotic diseases, including the Japanese encephalitis virus, Nipah virus, and influenza viruses. Several Orthomyxoviridae and Coronavirinae respiratory viruses require cleavage of envelope proteins to acquire viral infectivity and consequently, need a host protease or the addition of exogenous trypsin for efficient propagation. Host TMPRSS2 is a key protease responsible for viral cleavage. Stable expression of human TMPRSS2 in African green monkey-derived Vero cells can enhance the porcine epidemic diarrhea virus. However, considering the narrow host tropism of viruses, a porcine cell line expressing pig TMPRSS2 could be optimal for replicating pig-derived viruses. Herein, we generated and evaluated a pig-derived PK-15 cell line stably expressing pig TMPRSS2. This cell line markedly (>1000-fold) and specifically enhanced the growth of influenza viruses. Furthermore, we demonstrated the usefulness of a PK-15 cell line lacking the Stat2 gene with a stable expression of pig TMPRSS2 for efficient virus isolation from clinical samples in the presence of type I interferons. Therefore, PK-15 cells expressing pig TMPRSS2 could be a valuable and promising tool for virus isolation, vaccine production, and virological studies of TMPRSS2-dependent viruses.
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A case of hydronephrosis due to intrarenal ureteral obstruction in a Japanese Black calf Reviewed
Sato R., Hirai T., Hegazy A.A., Nakai M., Sato Y., Yamada K., Satoh H., Steiner A.
Journal of Veterinary Medical Science 86 ( 11 ) 1162 - 1167 2024
Publishing type：Research paper (scientific journal) Publisher：Journal of Veterinary Medical Science
A 23-day-old Japanese Black female calf presented with distension of the lower abdomen. Abdominal ultrasonography revealed a cystic structure with fluid accumulation and moderate echoluminance in the right abdominal cavity. Ultrasonography and contrast-enhanced computed tomography revealed congenital hydronephrosis due to narrowing of the intrarenal ureter, and right kidney resection was performed. Blood urea nitrogen and serum creatinine levels, which were within reference values preoperatively, peaked on the third postoperative day and gradually decreased afterward until reaching preoperative values on the tenth day. In bovine hydronephrosis, nephrectomy may provide a favorable prognosis if one kidney is intact. Moreover, this case suggests that blood tests immediately after surgery are not suitable for evaluating residual kidneys.
DOI： 10.1292/jvms.24-0173
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Surgical management of a dermoid cyst with mandibular gland tissue in a Japanese Black calf. Reviewed
Sato R, Sonoda M, Matsuo R, Takada R, Kanda I, Satoh H, Fuke N, Yamada K, Hirai T.
J Vet Med Sci. 19 ( 85 ) 1110 - 1115 2023.10
Language：English Publishing type：Research paper (scientific journal)
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Suwanruengsri M., Uemura R., Izzati U.Z., Kanda T., Fuke N., Yasuda M., Hirai T., Yamaguchi R.
Thai Journal of Veterinary Medicine 52 ( 1 ) 33 - 44 2022.3
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Thai Journal of Veterinary Medicine
Mycoplasma bovis (M. bovis) is a pathogen that especially effects the respiratory system of cattle. We focused on the histopathological changes in the lungs caused by M. bovis because the lesions in the bronchi or bronchioles and alveoli are quite different in natural infections. Thirty-five lung samples from 2-to 12-month-old Japanese Black calves with respiratory symptoms were collected and examined for bacteria presence and pathological changes using immunohistochemistry (IHC). Using the loop-mediated isothermal amplification method, 18 of the 35 samples were found to be positive for M. bovis, which was isolated and identified. In 7 of the samples, only M. bovis was detected, whereas in the other 11, other bacteria coexisted with M. bovis. Anti-M. bovis, anti-MAC387, anti-Iba1, anti-CD3, anti-CD20, anti-AE1/AE3 cytokeratin and anti-IL8 were used for antigen detection. Histopathological studies and diagnoses of the lungs were examined in two parts; bronchiolar lesions and alveolar lesions. The bronchiolar findings were divided into three groups: granulomatous, caseonecrotic and suppurative bronchiolitis. The alveolar lesions found were quite mild, except in three cases. IHC staining revealed strong evidence of M. bovis in degenerated neutrophils, which was confirmed by electron microscopy. The main inflammatory cell response to M. bovis infections was degenerate neutrophils, which caused the destruction of the bronchial epithelium and possibly induced epithelioid granulomatous inflammation, resulting in severe chronic respiratory diseases in cattle.
DOI： 10.14456/tjvm.2022.4
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Nueangphuet P., Tanabe M., Izzati U.Z., Fuke N., Hirai T., Yamaguchi R.
Journal of Veterinary Diagnostic Investigation 34 ( 1 ) 102 - 106 2022.1
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Journal of Veterinary Diagnostic Investigation
A 14-y-old Miniature Pinscher bitch was admitted to a veterinary clinic because of inappetence and a distended abdomen; ultrasound examination revealed a fluid-filled uterus with a single 1-cm hyperechoic nodule in its lumen. Ovariohysterectomy was performed. Grossly, the uterine horns were distended irregularly and asymmetrically, and the uterine lumen contained 200–300 mL of brown watery fluid. A single white polypoid 0.9-cm diameter nodule was present at the site of the distended uterine horn and arose from the endometrium with a narrow stalk. Histologically, the polyp consisted of dense, smooth muscle fascicles admixed with glandular components; its surface was covered by simple cuboidal epithelium with areas of squamous metaplasia. The myomatous cells expressed the myogenic markers smooth muscle actin and desmin. We interpreted the mass as an adenomyomatous uterine polyp, which is a rare variant of an endometrial polyp.
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Suwanruengsri M., Uemura R., Kanda T., Fuke N., Nueangphuet P., Pornthummawat A., Yasuda M., Hirai T., Yamaguchi R.
Journal of Veterinary Diagnostic Investigation 34 ( 1 ) 68 - 76 2022.1
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Journal of Veterinary Diagnostic Investigation
Mycoplasma bovis, the most important primary pathogen in the family Mycoplasmataceae, causes pneumonia, arthritis, otitis media, and mastitis in cattle. Histopathologic pulmonary changes associated with M. bovis infection have been characterized as suppurative-to-caseonecrotic bronchopneumonia; infection in other organs has been reported in only a few studies that examined caseonecrotic endocarditis and suppurative meningitis. Granulomatous lesions associated with M. bovis infection have been reported only rarely. We studied the granulomatous inflammation associated with M. bovis infection in several organs of 21 Japanese Black cattle. M. bovis was detected by isolation and loop-mediated isothermal amplification methods; other bacteria were detected using culture on 5% blood sheep agar and a MALDI-TOF MS Biotyper. Tissues were examined by histopathology and by immunohistochemistry (IHC) using anti–M. bovis, anti-Iba1, anti-iNOS, and anti-CD204 antibodies. All 21 cases, which included 2 cases of meningitis-meningoencephalitis, 8 cases of endocarditis, and 11 cases of bronchopneumonia, had caseonecrotic granulomatous inflammation associated with M. bovis infection. The IHC for macrophages revealed a predominance of iNOS-labeled (M1) macrophages in the inner layer of the caseonecrotic granulomas associated with meningitis-meningoencephalitis, endocarditis, and bronchopneumonia in Japanese Black cattle naturally infected with M. bovis.
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Toyama K., Hirai T., Sueyoshi M., Zhou Z., Niazi A.M., Kawaguchi N., Fuke N., Yamaguchi R.
Journal of Veterinary Medical Science 84 ( 11 ) 1536 - 1542 2022
Authorship：Corresponding author Publishing type：Research paper (scientific journal) Publisher：Journal of Veterinary Medical Science
Porcine circovirus 2 (PCV2) causes porcine circovirus-associated disease, and co-infection with porcine reproductive and respiratory syndrome virus (PRRSV) severely affects the pig breeding industry. Both viruses target the macrophages in lymphoid tissues. Various porcine pathogens enter via the nasal cavity, and the nasopharynx-associated lymphoid tissue (NALT) acts as the mucosal immune system. However, the pathological analysis has not progressed. This study aimed to histologically examine the NALT of pigs with suspected PCV2 and PRRSV infections. Six pigs were subjected to necropsy, and their NALT, tonsils, and mesenteric lymph nodes were collected. Macrophages, lymphocytic depletion, multinucleated giant cells, intracytoplasmic inclusion bodies, and neutrophil infiltration increased in the NALT. In situ hybridization revealed positive signals for PCV2 in the NALT of all pigs and PRRSV in the NALT of three pigs. PCV2-positive macrophages were mainly identified in the follicles, whereas PRRSV-positive tissues were found primarily around the crypt and directly below the epithelium. Quantitative PCR revealed 108 –1010 copies of PCV2 DNA/µL and 102 –104 copies of PRRSV DNA/µL in the NALT. Therefore, both PCV2 and PRRSV were detected in the NALT of pigs. In conclusion, the infection and replication of both viruses in the NALT and tonsils may suppress host immunity and promote co-infection with other pathogens.
DOI： 10.1292/jvms.22-0231
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Myint O., Hoa N.T., Fuke N., Pornthummawat A., Lan N.T., Hirai T., Yoshida A., Yamaguchi R.
BMC Veterinary Research 17 ( 1 ) 2021.12
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：BMC Veterinary Research
Background: Porcine epidemic diarrhoea (PED) is a highly contagious infectious disease with negative economic impacts on the swine industry. PED outbreaks were reported from 2009 to 2015, but sporadic infection has been observed until now in Vietnam. However, the seroprevalence of PEDV infection has not yet been reported for commercial pig farms in Vietnam. The aim of this study was to assess the seroprevalence of PEDV infection in Vietnamese pig farms to reveal the endemic status of PEDV in northern Vietnam. Results: A serological survey of PEDV infection was carried out using indirect ELISA in commercial pig farms in Hai Duong, Hung Yen and Thai Binh provinces in northern Vietnam in 2019. Twenty sera were randomly collected from each of 10 commercial pig farms, from each province; none of the farms had vaccinated for PEDV. Serological evidence of natural PEDV infection, expressed as a high antibody titre, was observed in the pig farms in all 3 provinces. The OD values were significantly higher (p < 0.001) for pig sera from Thai Binh than from Hai Duong and Hung Yen. No significant differences (p > 0.05) were detected for seropositivity to PEDV based on locality, age, pig breed and farm size. Conclusions: This study indicates serological evidence of natural PEDV infection with high antibody titre in commercial pig farms. PEDV infection was widespread among the pig population in these 3 provinces and that good management and strict biosecurity are needed at these pig farms.
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Nueangphuet P., Suwanruengsri M., Fuke N., Uemura R., Hirai T., Yamaguchi R.
Journal of Comparative Pathology 189 31 - 44 2021.11
Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Journal of Comparative Pathology
Mycoplasma hyopneumoniae (Mhp) is the primary pathogen of porcine enzootic pneumonia (PEP). Consolidated lung tissue from the cranioventral lung lobes of 15 pigs with PEP was collected for quantitative polymerase chain reaction, histopathology and immunohistochemistry. Histopathology revealed the co-existence of bronchial-associated lymphoid tissue hyperplasia with intra-alveolar neutrophils and macrophage infiltration in lesions of suppurative bronchopneumonia. Immunolabelling of infiltrated macrophages with CD163/CD204 indicated the presence of M2-polarized macrophages. Mhp antigen was detected on respiratory epithelial cells and in phagocytosed neutrophils. The intensity of Mhp immunolabelling and number of CD163/CD204-positive macrophages were correlated with the Mhp load in lung tissue (r = 0.87, 0.56, P <0.05). IL-8 immunolabelling was mainly found in neutrophils and correlated with Mhp load, Mhp immunolabelling and histological lesion score (r = 0.70, 0.66, 0.64, P <0.05), respectively. Apoptosis was seen in intra-alveolar cells and was correlated with Mhp load (r = 0.62, P <0.05). It is postulated that IL-8 attracts neutrophils to the lesions, while M2-polarized macrophages are a major source of IL-10 and promote a Th2-type immune response.