Molecular characterization and protective efficacy of the microneme 2 protein from Eimeria tenella

Microneme proteins play an important role in the adherence of apicomplexan parasites to host cells during the invasion process. In this study, the microneme 2 protein from the protozoan parasite Eimeria tenella (EtMIC2) was cloned, characterized, and its protective efficacy as a DNA vaccine investigated. The EtMIC2 gene, which codes for a 35.07 kDa protein in E. tenella sporulated oocysts, was cloned and recombinant EtMIC2 protein (rEtMIC2) was produced in an Escherichia coli expression system. Immunostaining with an anti-rEtMIC2 antibody showed that the EtMIC2 protein mainly localized in the anterior region and membrane of sporozoites, in the cytoplasm of first- and second-generation merozoites, and was strongly expressed during first-stage schizogony. In addition, incubation with specific antibodies against EtMIC2 was found to efficiently reduce the ability of E. tenella sporozoites to invade host cells. Furthermore, animal-challenge experiments demonstrated that immunization with pcDNA3.1(+)-EtMIC2 significantly increased average body weight gain, while decreasing the mean lesion score and oocyst output in chickens. Taken together, these results suggest that EtMIC2 plays an important role in parasite cell invasion and may be a viable candidate for the development of new vaccines against E. tenella infection in chickens.

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Genome-Wide Expression Patterns of Rhoptry Kinases during the Eimeria tenella Life-Cycle

Microorganisms ◽

10.3390/microorganisms9081621 ◽

2021 ◽

Vol 9 (8) ◽

pp. 1621

Author(s):

Adeline Ribeiro E Silva ◽

Alix Sausset ◽

Françoise I. Bussière ◽

Fabrice Laurent ◽

Sonia Lacroix-Lamandé ◽

...

Keyword(s):

Life Cycle ◽

Expression Patterns ◽

Housekeeping Genes ◽

Catalytic Triad ◽

Eimeria Tenella ◽

Apicomplexan Parasites ◽

Genome Wide ◽

Infected Cells ◽

First And Second Generation

Kinome from apicomplexan parasites is composed of eukaryotic protein kinases and Apicomplexa specific kinases, such as rhoptry kinases (ROPK). Ropk is a gene family that is known to play important roles in host–pathogen interaction in Toxoplasma gondii but is still poorly described in Eimeria tenella, the parasite responsible for avian coccidiosis worldwide. In the E. tenella genome, 28 ropk genes are predicted and could be classified as active (n = 7), inactive (incomplete catalytic triad, n = 12), and non-canonical kinases (active kinase with a modified catalytic triad, n = 9). We characterized the ropk gene expression patterns by real-time quantitative RT-PCR, normalized by parasite housekeeping genes, during the E. tenella life-cycle. Analyzed stages were: non-sporulated oocysts, sporulated oocysts, extracellular and intracellular sporozoites, immature and mature schizonts I, first- and second-generation merozoites, and gametes. Transcription of all those predicted ropk was confirmed. The mean intensity of transcription was higher in extracellular stages and 7–9 ropk were specifically transcribed in merozoites in comparison with sporozoites. Transcriptional profiles of intracellular stages were closely related to each other, suggesting a probable common role of ROPKs in hijacking signaling pathways and immune responses in infected cells. These results provide a solid basis for future functional analysis of ROPK from E. tenella.

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Exogenous nitric oxide stimulates early egress of Eimeria tenella sporozoites from primary chicken kidney cells in vitro

Parasite ◽

10.1051/parasite/2021007 ◽

2021 ◽

Vol 28 ◽

pp. 11

Author(s):

Xinlei Yan ◽

Wenying Han ◽

Xianyong Liu ◽

Xun Suo

Keyword(s):

Nitric Oxide ◽

Vital Role ◽

Eimeria Tenella ◽

Host Cells ◽

Host Immune System ◽

Kidney Cells ◽

Apicomplexan Parasites ◽

Immune Molecule ◽

Chicken Kidney

Egress plays a vital role in the life cycle of apicomplexan parasites including Eimeria tenella, which has been attracting attention from various research groups. Many recent studies have focused on early egress induced by immune molecules to develop a new method of apicomplexan parasite elimination. In this study, we investigated whether nitric oxide (NO), an immune molecule produced by different types of cells in response to cytokine stimulation, could induce early egress of eimerian sporozoites in vitro. Eimeria tenella sporozoites were extracted and cultured in primary chicken kidney cells. The number of sporozoites egressed from infected cells was analyzed by flow cytometry after treatment with NO released by sodium nitroferricyanide (II) dihydrate. The results showed that exogenous NO stimulated the rapid egress of E. tenella sporozoites from primary chicken kidney cells before replication of the parasite. We also found that egress was dependent on intra-parasitic calcium ion (Ca2+) levels and no damage occurred to host cells after egress. The virulence of egressed sporozoites was significantly lower than that of fresh sporozoites. The results of this study contribute to a novel field examining the interactions between apicomplexan parasites and their host cells, as well as that of the clearance of intracellular pathogens by the host immune system.

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Tampilan Produksi dan Efek Imunomodulasi Ayam Broiler yang Diberi Ransum Berbasis Wheat Pollard Terolah (PRODUCTION PERFORMANCE AND IMMUNOMODULATION EFFECTS ON BROILER GIVEN A PROCESSED WHEAT POLLARD BASED DIET)

jurnal veteriner ◽

10.19087/jveteriner.2019.20.3.352 ◽

2019 ◽

Vol 20 (3) ◽

pp. 352

Author(s):

Bambang Sulistiyanto ◽

Sri Kismiati ◽

Cahya Setya Utama

Keyword(s):

Body Weight ◽

Broiler Chickens ◽

Body Weight Gain ◽

Relative Weight ◽

Production Performance ◽

Metabolic Energy ◽

Feed Conversion ◽

Randomized Design ◽

The Mean ◽

Average Body

An immunomodulator is a natural body defence mechanism due to the stimulation of objects/ environments, both specific and non-specific by means of cellular and humoraldefence. The objective of the study was to prove the immunomodulation ability of broiler chickens up to age 35 days that dietary fed various wheat pollard based rations. The design used in the study was a complete randomized design with six treatments and three replications. The treatment of broiler rats consisted of BR-IAJ (T0), control feed with wheat pollard base (T1), control feed with wheat pollard base plus probiotic (T2), wheat pollard based feed steamed (T3), fermented wheat pollard based feed 40% (T4 ) and fermented wheat pollard based feed 60% (T5). The rations used contained 20,5-22,5% protein with 2900-3100 Kcal metabolic energy. Lohman MB 202 Platinum from PT. Japfa Comfeed used in research with average day old chick (DOC) weight of 36,39 + 2,45 g. Parameters observed included body weight, feed intake, feed conversion ratio (FCR), relative weights of liver organ, thymus, lymph and bursa fabricius. The results showed that wheat pollard-based rations had a significant effect (p <0,05) on the relative weight of liver organ, body weight, FCR and ration consumption, but did not affect the relative weights of spleen organ, thymus and bursa fabricius. The mean relative weight of consecutive liver as follows: 2,47 g (T0), 2,83 g (T1), 3,40 g (T2), 2,99 g (T3), 3,15 g (T4) and 3,06 g (T5). The average body weight, consumption and FCR rations were as follows: 1415,24 g; 136,45 g/day; 2,03 (T0), 775,04 g; 125,19 g/day; 3,40 (T1), 813,17 g; 129,86 g / day; 3,36 (T2), 792,89 g; 128,14 g /day; 3,54 (T3), 892,91 g; 138,33 g /day; 3,27 (T4), 969,56 g; 155,52 g/day; 3,37 (T5). The conclusion of the study was that the ration with the addition of wheat pollard 60% was able to provide an increase in body weight gain and the best immunomodulation seen from the aspect of the proportion of immunomodulating organs to the growth of broiler chickens.

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Molecular characterization and protective efficacy of a new conserved hypothetical protein of Eimeria tenella

Parasite ◽

10.1051/parasite/2021037 ◽

2021 ◽

Vol 28 ◽

pp. 40

Author(s):

Huanzhi Zhao ◽

Shunhai Zhu ◽

Qiping Zhao ◽

Bing Huang ◽

Guiling Liu ◽

...

Keyword(s):

Western Blot ◽

Second Generation ◽

Protective Efficacy ◽

Hypothetical Protein ◽

Eimeria Tenella ◽

Host Cells ◽

Rabbit Serum ◽

Pcr Analysis ◽

Mrna Levels ◽

A Genome

Eimeria tenella is an obligate intracellular parasite that actively invades cecal epithelial cells of chickens. This parasite encodes a genome of more than 8000 genes. However, more than 70% of the gene models for this species are currently annotated as hypothetical proteins. In this study, a conserved hypothetical protein gene of E. tenella, designated EtCHP18905, was cloned and identified, and its immune protective effects were evaluated. The open reading frame of EtCHP18905 was 1053bp and encoded a protein of 350 amino acids with a molecular weight of 38.7kDa. The recombinant EtCHP18905 protein (rEtCHP18905) was expressed in E. coli. Using western blot, the recombinant protein was successfully recognized by anti GST-Tag monoclonal antibody and anti-sporozoites protein rabbit serum. Real-time quantitative PCR analysis revealed that the EtCHP18905 mRNA levels were higher in sporozoites than in unsporulated oocysts, sporulated oocysts and second-generation merozoites. Western blot analysis showed that EtCHP18905 protein expression levels were lower in sporozoites than in other stages. Immunofluorescence analysis indicated that the EtCHP18905 protein was located on the surface of sporozoites and second-generation merozoites. Inhibition experiments showed that the ability of sporozoites to invade host cells was significantly decreased after treatment with the anti-rEtCHP18905 polyclonal antibody. Vaccination with rEtCHP18905 protein was able to significantly decrease mean lesion scores and oocyst outputs as compared to non-vaccinated controls. The results suggest that the rEtCHP18905 protein can induce partial immune protection against infection with E. tenella and could be an effective candidate for the development of new vaccines.

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A Novel Rhoptry Protein as Candidate Vaccine against Eimeria tenella Infection

Vaccines ◽

10.3390/vaccines8030452 ◽

2020 ◽

Vol 8 (3) ◽

pp. 452

Author(s):

Xingju Song ◽

Xu Yang ◽

Taotao Zhang ◽

Jing Liu ◽

Qun Liu

Keyword(s):

Body Weight ◽

Weight Gain ◽

Survival Rate ◽

Protective Efficacy ◽

Body Weight Gain ◽

Surface Antigens ◽

Eimeria Tenella ◽

Dense Granule ◽

Control Group ◽

Lesion Score

Eimeria tenella (E. tenella) is a highly pathogenic and prevalent species of Eimeria that infects chickens, and it causes a considerable disease burden worldwide. The secreted proteins and surface antigens of E. tenella at the sporozoite stage play an essential role in the host–parasite interaction, which involves attachment and invasion, and these interactions are considered vaccine candidates based on the strategy of cutting off the invasion pathway to interrupt infection. We selected two highly expressed surface antigens (SAGs; Et-SAG13 and Et-SAG) and two highly expressed secreted antigens (rhoptry kinases Eten5-A, Et-ROPK-Eten5-A and dense granule 12, Et-GRA12) at the sporozoite stage. Et-ROPK-Eten5-A and Et-GRA12 were two unexplored proteins. Et-ROPK-Eten5-A was an E. tenella-specific rhoptry (ROP) protein and distributed in the apical pole of sporozoites and merozoites. Et-GRA12 was scattered in granular form at the sporozoite stage. To evaluate the potential of rEt-ROPK-Eten5-A, rEt-GRA12, rEt-SAG13 and rEt-SAG proteins as a coccidiosis vaccine, the protective efficacy was examined based on survival rate, lesion score, body weight gain, relative body weight gain and oocyst output. The survival rate was significantly improved in rEt-ROPK-Eten5-A (100%) and rEt-GRA12 (100%) immune chickens compared to the challenged control group (40%). The average body weight gains of rEt-ROPK-Eten5-A, rEt-GRA12, rEt-SAG13 and rEt-SAG immunized chickens were significantly higher than those of unimmunized chickens. The mean lesion score and oocyst output of the rEt-ROPK-Eten5-A immunized chickens were significantly reduced compared to unimmunized challenged chickens. These results suggest that the rEt-ROPK-Eten5-A protein effectively triggered protection against E. tenella in chickens and provides a useful foundation for future work developing anticoccidial vaccines.

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Genetic and phenotypic parameter estimates for body weight and egg production at sexual maturity in Hilly×Fayoumi crossbred chickens

Asian Journal of Medical and Biological Research ◽

10.3329/ajmbr.v4i2.38254 ◽

2018 ◽

Vol 4 (2) ◽

pp. 186-192

Author(s):

Ashutosh Das ◽

Mukta Das Gupta ◽

Md Kabirul Islam Khan ◽

Md Moksedul Momin ◽

Omar Faruk Miazi

Keyword(s):

Body Weight ◽

Sexual Maturity ◽

Egg Production ◽

High Weight ◽

Body Weight Gain ◽

Parameter Estimates ◽

Indigenous Chicken ◽

Chicken Breed ◽

The Mean ◽

Average Body

A crossbreeding experiment between Hilly, a locally adapted chicken type in Bangladesh and Fayoumi, an egg type light chicken breed was carried out to evaluate phenotypic performances and to estimate genetic parameters for body weight and egg production at sexual maturity. Results show the mean hatch weight of Hilly♂ × Fayoumi♀ (H♂×F♀) crosses was 35±0.1 gm. In this study, H♂×F♀ crossbred showed a heavier body weight (1551± 32.0 gm/bird) at 20 weeks in comparison with other indigenous chicken genotypes in Bangladesh. The highest body weight gain was found in 8-10 weeks of age. H♂×F♀ crossbred hens exhibited sexual maturity at an average age of 147.5±1.6 days with an average body weight of 1350± 16.8 gm/bird. The mean weight of eggs at sexual maturity was 33.7±0.5 gm/egg. The estimates of heritability for body weight were ranged from 0.15 to 0.26. We observed a positive genetic correlation between weight at sexual maturity (WSM) and egg weight at sexual maturity (EWSM), meaning that hens with high weight at sexual maturity would produce heavier eggs.Asian J. Med. Biol. Res. June 2018, 4(2): 186-192

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Calcium-dependent phosphorylation alters class XIVa myosin function in the protozoan parasite Toxoplasma gondii

Molecular Biology of the Cell ◽

10.1091/mbc.e13-11-0648 ◽

2014 ◽

Vol 25 (17) ◽

pp. 2579-2591 ◽

Cited By ~ 26

Author(s):

Qing Tang ◽

Nicole Andenmatten ◽

Miryam A. Hortua Triana ◽

Bin Deng ◽

Markus Meissner ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Host Cell ◽

Calcium Ionophore ◽

Protozoan Parasite ◽

Gliding Motility ◽

Lytic Cycle ◽

Host Cells ◽

Response To Treatment ◽

Apicomplexan Parasites ◽

Calcium Dependent

Class XIVa myosins comprise a unique group of myosin motor proteins found in apicomplexan parasites, including those that cause malaria and toxoplasmosis. The founding member of the class XIVa family, Toxoplasma gondii myosin A (TgMyoA), is a monomeric unconventional myosin that functions at the parasite periphery to control gliding motility, host cell invasion, and host cell egress. How the motor activity of TgMyoA is regulated during these critical steps in the parasite's lytic cycle is unknown. We show here that a small-molecule enhancer of T. gondii motility and invasion (compound 130038) causes an increase in parasite intracellular calcium levels, leading to a calcium-dependent increase in TgMyoA phosphorylation. Mutation of the major sites of phosphorylation altered parasite motile behavior upon compound 130038 treatment, and parasites expressing a nonphosphorylatable mutant myosin egressed from host cells more slowly in response to treatment with calcium ionophore. These data demonstrate that TgMyoA undergoes calcium-dependent phosphorylation, which modulates myosin-driven processes in this important human pathogen.

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Transcriptional dynamics in the protozoan parasite Sarcocystis neurona and mammalian host cells after treatment with a specific inhibitor of apicomplexan mRNA polyadenylation

PLoS ONE ◽

10.1371/journal.pone.0259109 ◽

2021 ◽

Vol 16 (10) ◽

pp. e0259109

Author(s):

Arthur G. Hunt ◽

Daniel K. Howe ◽

Ashley Brown ◽

Michelle Yeargan

Keyword(s):

Marine Mammals ◽

Specific Inhibitor ◽

Protozoan Parasite ◽

Host Cells ◽

Mammalian Host ◽

Sarcocystis Neurona ◽

Apicomplexan Parasites ◽

Or Gene ◽

A Site ◽

Mrna Polyadenylation

In recent years, a class of chemical compounds (benzoxaboroles) that are active against a range of parasites has been shown to target mRNA polyadenylation by inhibiting the activity of CPSF73, the endonucleolytic core of the eukaryotic polyadenylation complex. One particular compound, termed AN3661, is active against several apicomplexan parasites that cause disease in humans. In this study, we report that AN3661 is active against an apicomplexan that causes disease in horses and marine mammals (Sarcocystis neurona), with an approximate IC50 value of 14.99 nM. Consistent with the reported mode of action of AN3661 against other apicomplexans, S. neurona mutants resistant to AN3661 had an alteration in CPSF73 that was identical to a mutation previously documented in AN3661-resistant Toxoplasma gondii and Plasmodium falciparum. AN3661 had a wide-ranging effect on poly(A) site choice in S. neurona, with more than half of all expressed genes showing some alteration in mRNA 3’ ends. This was accompanied by changes in the relative expression of more than 25% of S. neurona genes and an overall 5-fold reduction of S. neurona transcripts in infected cells. In contrast, AN3661 had no discernible effect on poly(A) site choice or gene expression in the host cells. These transcriptomic studies indicate that AN3661 is exceedingly specific for the parasite CPSF73 protein, and has the potential to augment other therapies for the control of apicomplexan parasites in domestic animals.

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THE EFFECT OF (Morinda citrifolia L.) MEAL IN DIET ON PERFORMANCE OF BROILER

Jurnal Agroindustri ◽

10.31186/j.agroind.2.1.21-27 ◽

2012 ◽

Vol 2 (1) ◽

pp. 21-27

Author(s):

Yosi Fenita

Keyword(s):

Body Weight ◽

Body Weight Gain ◽

Morinda Citrifolia ◽

Feed Consumption ◽

Feed Conversion ◽

Randomized Design ◽

Different Levels ◽

Effect Of Feeding ◽

Average Body ◽

Average Body Weight

The objective of the research was to evaluate to effect of feeding mengkudu on performances of broilers. The research design used was completely randomized design. One hundred broilers were distributed into five treatments. The treatments were different levels of mengkudu meal (0, 0.75%, 1.5%, 2.25 % and 3%). The observed measured were feed consumption, average body weight (gain) and feed conversion. Results showed that feeding mengkudu (Morinda Citrifolia L.) no effect significant (P>0.05) on feed consumption, average body weight and feed conversion. In conclusion, feeding mengkudu meal up to 3% (in diet) does not negatively affect feed consumption, average body weight, and feed conversion.

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Development of a Viral-Like Particle Candidate Vaccine Against Novel Variant Infectious Bursal Disease Virus

Vaccines ◽

10.3390/vaccines9020142 ◽

2021 ◽

Vol 9 (2) ◽

pp. 142

Author(s):

Yulong Wang ◽

Nan Jiang ◽

Linjin Fan ◽

Li Gao ◽

Kai Li ◽

...

Keyword(s):

Disease Virus ◽

Infectious Bursal Disease Virus ◽

Protective Efficacy ◽

Expression System ◽

Size Exclusion ◽

Candidate Vaccine ◽

Infectious Bursal Disease ◽

Immune Protection ◽

Bursal Disease ◽

Novel Variant

Infectious bursal disease (IBD), an immunosuppressive disease of young chickens, is caused by infectious bursal disease virus (IBDV). Novel variant IBDV (nVarIBDV), a virus that can evade immune protection against very virulent IBDV (vvIBDV), is becoming a threat to the poultry industry. Therefore, nVarIBDV-specific vaccine is much needed for nVarIBDV control. In this study, the VP2 protein of SHG19 (a representative strain of nVarIBDV) was successfully expressed using an Escherichia coli expression system and further purified via ammonium sulfate precipitation and size-exclusion chromatography. The purified protein SHG19-VP2-466 could self-assemble into 25-nm virus-like particle (VLP). Subsequently, the immunogenicity and protective effect of the SHG19-VLP vaccine were evaluated using animal experiments, which indicated that the SHG19-VLP vaccine elicited neutralization antibodies and provided 100% protection against the nVarIBDV. Furthermore, the protective efficacy of the SHG19-VLP vaccine against the vvIBDV was evaluated. Although the SHG19-VLP vaccine induced a comparatively lower vvIBDV-specific neutralization antibody titer, it provided good protection against the lethal vvIBDV. In summary, the SHG19-VLP candidate vaccine could provide complete immune protection against the homologous nVarIBDV as well as the heterologous vvIBDV. This study is of significance to the comprehensive prevention and control of the recent atypical IBD epidemic.

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