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Identification and Characterization of α-Actinin 1 of Histomonas meleagridis and Its Potential Vaccine Candidates against Histomonosis.

ABSTRACT: Histomonas meleagridis is a protozoan parasite that causes histomonosis in gallinaceous birds such as turkeys and chickens. Since the banning and restricted usage of effective drugs such as nitarsone, 80-100% morbidity and mortality occur in turkeys and 20-30% mortality in chickens. New ideas are needed to resolve the re-emergence of histomonosis in poultry. In this study, the α-actinin encoding gene from H. meleagridis was cloned. The 1839-bp gene encoding 612 amnio acids showed close phylogenetic relationships with Trichomonas vaginalis and Tritrichomonas foetus. It was then inserted into the prokaryotic expression vector pET28a(+) and induced with isopropyl-β-D-thiogalactopyranoside. A 73 kDa recombinant protein rHmα-actinin 1 was obtained and purified with a Ni-NTA chromatography column. rHmα-actinin 1 was recognized by mouse anti-rHmα-actinin 1 polyclonal antibody, mouse anti-rHmα-actinin 1 monoclonal antibody, and rehabilitation sera from H. meleagridis infected chickens. Native α-actinin 1 in the total proteins of H. meleagridis can also be detected with mouse anti-rHmα-actinin monoclonal antibody. Immunolocalization assays showed that Hmα-actinin 1 was mainly distributed in the cytoplasm of virulent histomonads JSYZ-D9 and in the peripheral regions (near the plasma membrane) of attenuated histomonads JSYZ-D195. Based on in vivo experiment, when chickens were subcutaneously immunized with rHmα-actinin 1 at 5 and 12 days old and then challenged with H. meleagridis at 19 days old, rHmα-actinin 1 reduced the lesion scores 12 days after infection (31 days old) and increased the body weight gain during the challenged period (19-31 days old). Furthermore, it also strengthened the cellular and humoral immune responses 7 days after the second immunization (19 days old). In conclusion, Hmα-actinin 1 could be used as a candidate antigen to develop vaccines against chicken histomonosis.

SUBMITTER: Liu D

PROVIDER: S-EPMC10376378 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature

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Identification and Characterization of α-Actinin 1 of Histomonas meleagridis and Its Potential Vaccine Candidates against Histomonosis.

Liu Dandan D Chen Chen C Chen Qiaoguang Q Wang Shuang S Li Zaifan Z Rong Jie J Zhang Yuming Y Hou Zhaofeng Z Tao Jianping J Xu Jinjun J

Animals : an open access journal from MDPI 20230717 14

Histomonas meleagridis is a protozoan parasite that causes histomonosis in gallinaceous birds such as turkeys and chickens. Since the banning and restricted usage of effective drugs such as nitarsone, 80-100% morbidity and mortality occur in turkeys and 20-30% mortality in chickens. New ideas are needed to resolve the re-emergence of histomonosis in poultry. In this study, the α-actinin encoding gene from H. meleagridis was cloned. The 1839-bp gene encoding 612 amnio acids showed c ...[more]

PMID: 37508107

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Project description:IntroductionHistomonosis, a protozoan disease caused by Histomonas meleagridis, poses a significant economic burden on domestic poultry in China. To reduce the losses caused by this disease in chickens, an attenuated vaccine was developed by exploiting the diminished virulence of H. meleagridis through successive in vitro passages.MethodsFour experiments were conducted to evaluate the viability of attenuated H. meleagridis as a potential vaccine candidate. Experiment 1 evaluated the route of infection (oral vs. intracloacal) and dose (5 × 104, 1 × 105, and 2 × 105 H. meleagridis/chicken) using the virulent strain H. meleagridis JSYZ-D10. Experiment 2 evaluated the attenuated effect of the H. meleagridis JSYZ-D168 strain (infection dose: 2 × 105 H. meleagridis/chicken). Experiment 3 evaluated the immunoprotective effect of different immunization doses (5 × 104, 1 × 105, and 2 × 105 H. meleagridis/chicken). Experiment 4 evaluated the immunoprotective effect of different immunization schedules (immunization at 3 days of age; immunization at 14 days of age; two immunizations, one at 3 days of age and one at 14 days of age; immunization and infection dose: 2 × 105 H. meleagridis/chicken).ResultsThe results showed that the intracloacal route of infection was more effective and stable compared to the oral route. The pathogenicity of the JSYZ-D168 H. meleagridis strain was significantly reduced compared to the original virulent strain. Chickens vaccinated by intracloacal immunization at a dose of 2 × 105 H. meleagridis/chicken on day 14 provided effective protection against a virulent strain challenge, significantly resulting in increased body weight and reduced lesions in the cecum and liver within 28 days post-immunization (p < 0.05). Poor immunoprotection was obtained either when the immunization dose was 1 × 105 H. meleagridis/chicken or when the immunization program was a single immunization at 3 days of age only.DiscussionIn conclusion, the administration of a vaccine provides a measurable degree of protection against the detrimental effects induced by H. meleagridis, thus warranting its endorsement in clinical settings.

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Identification and Characterization of α-Actinin 1 of Histomonas meleagridis and Its Potential Vaccine Candidates against Histomonosis.

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Identification and Characterization of α-Actinin 1 of <i>Histomonas meleagridis</i> and Its Potential Vaccine Candidates against Histomonosis.

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