Pub Date : 2019-10-14DOI: 10.25431/1824-307X/ISJ.V0I0.164-172
X. Cui, J. W. Cheng, Yan Wang, B. Zhou
The present study describes the molecular characterization and transcriptional features of a tumor suppressor QM protein gene, MmQM, in Meretrix meretrix. The full-length cDNA (819 bp) of MmQM consists of a 657 bp opening reading frame (ORF) encoding a 218 amino acid protein with a calculated molecular mass of 25.3 kDa and theoretical isoelectric point of 10.2. A ribosomal protein L10 signature, an SH3-binding motif, an antibiotic binding site, an amidation site and two protein kinase C phosphorylation sites were revealed from the MmQM sequence. Phylogenetic analysis showed that MmQM is clustered with previously identified mollusk QM proteins. MmQM mRNA transcripts were detectable in all of the examined tissues in a constitutive manner, and were significantly different from each other. After bacterial stimulation, the mRNA transcripts of MmQM in the hepatopancreas significantly increased. Hence, we conclude that MmQM could respond to pathogenic infections and it might play an important role in the innate immunity against microorganisms in the clam M. meretrix.
{"title":"Sequence feature and expression profile of a tumor suppressor QM protein gene from hard clam Meretrix meretrix","authors":"X. Cui, J. W. Cheng, Yan Wang, B. Zhou","doi":"10.25431/1824-307X/ISJ.V0I0.164-172","DOIUrl":"https://doi.org/10.25431/1824-307X/ISJ.V0I0.164-172","url":null,"abstract":"The present study describes the molecular characterization and transcriptional features of a tumor suppressor QM protein gene, MmQM, in Meretrix meretrix. The full-length cDNA (819 bp) of MmQM consists of a 657 bp opening reading frame (ORF) encoding a 218 amino acid protein with a calculated molecular mass of 25.3 kDa and theoretical isoelectric point of 10.2. A ribosomal protein L10 signature, an SH3-binding motif, an antibiotic binding site, an amidation site and two protein kinase C phosphorylation sites were revealed from the MmQM sequence. Phylogenetic analysis showed that MmQM is clustered with previously identified mollusk QM proteins. MmQM mRNA transcripts were detectable in all of the examined tissues in a constitutive manner, and were significantly different from each other. After bacterial stimulation, the mRNA transcripts of MmQM in the hepatopancreas significantly increased. Hence, we conclude that MmQM could respond to pathogenic infections and it might play an important role in the innate immunity against microorganisms in the clam M. meretrix.","PeriodicalId":14623,"journal":{"name":"ISJ-Invertebrate Survival Journal","volume":null,"pages":null},"PeriodicalIF":0.7,"publicationDate":"2019-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75325419","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-10-14DOI: 10.25431/1824-307X/ISJ.V0I0.173-183
Qingzhong Liu, B. Xu, T. Xiao
Serine proteases play central roles in immune defense in invertebrates through innate immunity, and are particularly important complement system in molluscs because their susceptibility to infection due to lack of an adaptive immune ability. A gene encoding the serine protease homolog from the triangle - shell pearl mussel (Hyriopsis cumingii) was identified and designated as HC-SPH in this study. Protein sequence analysis revealed that HC-SPH consists of a typical Tryp_SPc functional domain of serine protease of S1 family lead by a signal peptide, and the molecule shares a highly conserved sequence and structural organization with other members, including a cleavage site, 3 enzymatic active sites and 3 substrate binding sites, so that it was clustered into a trypsin-like serine protease subfamily of the S1 superfamily. Semi - quantitative analysis of the amplicons separated on agarose gel by comparing to the β-actin products revealed that the digestive gland had a strong expression while the gonads were seen as weak expression sites. Infected by Aeromonas hydrophila, the gene expression was significantly up - regulated in the kidney at the 6 hours post challenge (hpc), stomach at 12 hpc and gills at 24 hpc while the expression maintained steadily unchanged in the digestive gland. However, up to 48 hpc, the expression levels in all four tissues reached significantly high, and also joined by a high level of expression in intestine that was down - regulated before 24 hpc, to build up an enhanced immune defense. The complementary up - regulation of the gene expression in these tissues suggested a temporal and spatial reinforce model for HC-SPH in immune response.
{"title":"HC-SPH: A conserved serine protease homolog of S1 superfamily in the triangle-shell pearl mussel (Hyriopsis cumingii)","authors":"Qingzhong Liu, B. Xu, T. Xiao","doi":"10.25431/1824-307X/ISJ.V0I0.173-183","DOIUrl":"https://doi.org/10.25431/1824-307X/ISJ.V0I0.173-183","url":null,"abstract":"Serine proteases play central roles in immune defense in invertebrates through innate immunity, and are particularly important complement system in molluscs because their susceptibility to infection due to lack of an adaptive immune ability. A gene encoding the serine protease homolog from the triangle - shell pearl mussel (Hyriopsis cumingii) was identified and designated as HC-SPH in this study. Protein sequence analysis revealed that HC-SPH consists of a typical Tryp_SPc functional domain of serine protease of S1 family lead by a signal peptide, and the molecule shares a highly conserved sequence and structural organization with other members, including a cleavage site, 3 enzymatic active sites and 3 substrate binding sites, so that it was clustered into a trypsin-like serine protease subfamily of the S1 superfamily. Semi - quantitative analysis of the amplicons separated on agarose gel by comparing to the β-actin products revealed that the digestive gland had a strong expression while the gonads were seen as weak expression sites. Infected by Aeromonas hydrophila, the gene expression was significantly up - regulated in the kidney at the 6 hours post challenge (hpc), stomach at 12 hpc and gills at 24 hpc while the expression maintained steadily unchanged in the digestive gland. However, up to 48 hpc, the expression levels in all four tissues reached significantly high, and also joined by a high level of expression in intestine that was down - regulated before 24 hpc, to build up an enhanced immune defense. The complementary up - regulation of the gene expression in these tissues suggested a temporal and spatial reinforce model for HC-SPH in immune response.","PeriodicalId":14623,"journal":{"name":"ISJ-Invertebrate Survival Journal","volume":null,"pages":null},"PeriodicalIF":0.7,"publicationDate":"2019-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87939851","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-11DOI: 10.25431/1824-307X/ISJ.V0I0.152-163
J. Auclair, C. André, C. Peyrot, K. Wilkinson, P. Turcotte, C. Gagnon, F. Gagné
Cerium oxide nanoparticles (nCeO) are currently used in many sectors of our economy, for instance as fuel additives and in ceramics for catalytic converters. As a result, there are concerns about their release and resulting toxicity in the aquatic environment. The purpose of this study was to examine the bioavailability and toxicity of nCeO and Ce(IV) in zebra mussels (Dreissena polymorpha) in various types of surface water differing in organic matter, conductivity and pH. Mussels were exposed to 100 μg/L Ce as either nCeO or Ce(IV) for 96 h in 4 types of water: 1) green water (high conductivity and low total organic carbon), 2) brown water (low conductivity and high natural total organic matter), 3) 10% municipal effluent (high conductivity and high anthropogenic organic matter) and 4) controls, which consisted of dechlorinated tap water. After the exposure period, the mussels were analyzed for morphological changes, resistance to survive in air, triglycerides (fat reserves), oxidative stress (arachidonate cyclooxygenase and lipid peroxidation) and DNA damage. Evidence of aggregation was observed with nCeO in most types of water, with the exception of the diluted municipal effluent. The data revealed that some of the effects of nCeO were influenced by surface water properties. The mussels were more sensitive to air emersion when exposed to nCeO in green water but not in the other water types and Ce(IV) to all types of water, although a marginal decrease was observed in mussels co-exposed to the diluted municipal effluent. A general decrease in oxidative stress and lipid levels was observed with both forms of Ce and all water types. Ce(IV) in brown water did not reduce the levels of DNA strand breaks compared with the controls. In conclusion, the sublethal toxicity of nCeO could be modulated by the surface water from which the nanoparticle is suspended.
{"title":"Combined effects of surface waters and CeO nanoparticle in zebra mussels","authors":"J. Auclair, C. André, C. Peyrot, K. Wilkinson, P. Turcotte, C. Gagnon, F. Gagné","doi":"10.25431/1824-307X/ISJ.V0I0.152-163","DOIUrl":"https://doi.org/10.25431/1824-307X/ISJ.V0I0.152-163","url":null,"abstract":"Cerium oxide nanoparticles (nCeO) are currently used in many sectors of our economy, for instance as fuel additives and in ceramics for catalytic converters. As a result, there are concerns about their release and resulting toxicity in the aquatic environment. The purpose of this study was to examine the bioavailability and toxicity of nCeO and Ce(IV) in zebra mussels (Dreissena polymorpha) in various types of surface water differing in organic matter, conductivity and pH. Mussels were exposed to 100 μg/L Ce as either nCeO or Ce(IV) for 96 h in 4 types of water: 1) green water (high conductivity and low total organic carbon), 2) brown water (low conductivity and high natural total organic matter), 3) 10% municipal effluent (high conductivity and high anthropogenic organic matter) and 4) controls, which consisted of dechlorinated tap water. After the exposure period, the mussels were analyzed for morphological changes, resistance to survive in air, triglycerides (fat reserves), oxidative stress (arachidonate cyclooxygenase and lipid peroxidation) and DNA damage. Evidence of aggregation was observed with nCeO in most types of water, with the exception of the diluted municipal effluent. The data revealed that some of the effects of nCeO were influenced by surface water properties. The mussels were more sensitive to air emersion when exposed to nCeO in green water but not in the other water types and Ce(IV) to all types of water, although a marginal decrease was observed in mussels co-exposed to the diluted municipal effluent. A general decrease in oxidative stress and lipid levels was observed with both forms of Ce and all water types. Ce(IV) in brown water did not reduce the levels of DNA strand breaks compared with the controls. In conclusion, the sublethal toxicity of nCeO could be modulated by the surface water from which the nanoparticle is suspended.","PeriodicalId":14623,"journal":{"name":"ISJ-Invertebrate Survival Journal","volume":null,"pages":null},"PeriodicalIF":0.7,"publicationDate":"2019-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82910797","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-08-01DOI: 10.25431/1824-307X/ISJ.V0I0.130-140
S. Kausar, M. N. Abbas, Yongju Zhao, H. Cui
The silkworm, Bombyx mori has great economic and scientific value, as it has long been exploited as a primary silk producer and as a model system for lepidopterans and arthropod studies. This species is highly susceptible to microbial diseases that affect quality and quantity of silk, thereby causing huge economical losses. Insects have developed efficient innate immune system to fight against microbial pathogens. The innate immune system plays a crucial biological role in the limitation of microbial infections by using different immune strategies such as antimicrobial peptides production (AMPs), reactive oxygen species generation and melanin formation. So far, many studies identified different biological factors, which are considered to be involve in the regulation of these biochemical processes in B. mori. Here, we describe, current knowledge on the molecular patterns of various immune factors and also highlight their molecular mechanism of action in the limitation of viral, bacterial and fungal pathogens in B. mori. Furthermore, we discussed different strategies to improve the immune responses of silkworm species. This review will be helpful to understand the molecular aspects of immune factors, and their regulatory mechanism to control microbial diseases in the economically important insect species, B. mori.
{"title":"Immune strategies of silkworm, Bombyx mori against microbial infections","authors":"S. Kausar, M. N. Abbas, Yongju Zhao, H. Cui","doi":"10.25431/1824-307X/ISJ.V0I0.130-140","DOIUrl":"https://doi.org/10.25431/1824-307X/ISJ.V0I0.130-140","url":null,"abstract":"The silkworm, Bombyx mori has great economic and scientific value, as it has long been exploited as a primary silk producer and as a model system for lepidopterans and arthropod studies. This species is highly susceptible to microbial diseases that affect quality and quantity of silk, thereby causing huge economical losses. Insects have developed efficient innate immune system to fight against microbial pathogens. The innate immune system plays a crucial biological role in the limitation of microbial infections by using different immune strategies such as antimicrobial peptides production (AMPs), reactive oxygen species generation and melanin formation. So far, many studies identified different biological factors, which are considered to be involve in the regulation of these biochemical processes in B. mori. Here, we describe, current knowledge on the molecular patterns of various immune factors and also highlight their molecular mechanism of action in the limitation of viral, bacterial and fungal pathogens in B. mori. Furthermore, we discussed different strategies to improve the immune responses of silkworm species. This review will be helpful to understand the molecular aspects of immune factors, and their regulatory mechanism to control microbial diseases in the economically important insect species, B. mori.","PeriodicalId":14623,"journal":{"name":"ISJ-Invertebrate Survival Journal","volume":null,"pages":null},"PeriodicalIF":0.7,"publicationDate":"2019-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78382679","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-07-11DOI: 10.25431/1824-307X/ISJ.V0I0.120-129
M. Mallahi, A. Zibaee, Jalal Jalali Sendi, S. Jamali
The effect of Steinernema carpocapsae (IRA18) infection was studied on the mortality, immunity and antioxidant responses of Glyphodes pyloalis Walker larvae. The LC50 value of 582.9 infective juvenile per mL with confidence limit of 359.1-811.5 was obtained via bioassay against the larvae. Injection of S. carpocapsae increased the number of total hemocytes after 1-6 h compared to intact and Ringer-injected larvae while the highest numbers of plasmatocyte and granulocytes were recorded after 1 and 3 h. Although intact larvae had a steady activity of phenoloxidase at different time intervals but those injected by S. carpocapsae showed the elevated enzymatic activity at 3-12 h. Nematode injection significantly increased the activities of superoxide dismutase and catalase compared to intact and Ringer-injected larvae, while no significant difference was observed in peroxidase activity. The injection with S. carpocapsae caused the highest activity of glutathione s-transferase using CDNB as reagent, but the enzymatic assay with DCNB showed no statistical differences among treatments. Also, activities of ascorbate peroxidase and glucose-6-phosphate dehydrogenase significantly increased in the nematode-injected larvae. Intact and Ringer-injected larvae showed no statistical differences in the concentration of malondialdehyde but the highest amount was recorded in nematode-injected larvae. Results of our study indicate that native isolate of S. carpocapsae cause mortality on the larvae of G. pyloalis and It interferes in the immune and antioxidant responses.
{"title":"Effects of Steinernema carpocapsae (Weiser) on immunity and antioxidant responses of Glyphodes pyloalis Walker","authors":"M. Mallahi, A. Zibaee, Jalal Jalali Sendi, S. Jamali","doi":"10.25431/1824-307X/ISJ.V0I0.120-129","DOIUrl":"https://doi.org/10.25431/1824-307X/ISJ.V0I0.120-129","url":null,"abstract":"The effect of Steinernema carpocapsae (IRA18) infection was studied on the mortality, immunity and antioxidant responses of Glyphodes pyloalis Walker larvae. The LC50 value of 582.9 infective juvenile per mL with confidence limit of 359.1-811.5 was obtained via bioassay against the larvae. Injection of S. carpocapsae increased the number of total hemocytes after 1-6 h compared to intact and Ringer-injected larvae while the highest numbers of plasmatocyte and granulocytes were recorded after 1 and 3 h. Although intact larvae had a steady activity of phenoloxidase at different time intervals but those injected by S. carpocapsae showed the elevated enzymatic activity at 3-12 h. Nematode injection significantly increased the activities of superoxide dismutase and catalase compared to intact and Ringer-injected larvae, while no significant difference was observed in peroxidase activity. The injection with S. carpocapsae caused the highest activity of glutathione s-transferase using CDNB as reagent, but the enzymatic assay with DCNB showed no statistical differences among treatments. Also, activities of ascorbate peroxidase and glucose-6-phosphate dehydrogenase significantly increased in the nematode-injected larvae. Intact and Ringer-injected larvae showed no statistical differences in the concentration of malondialdehyde but the highest amount was recorded in nematode-injected larvae. Results of our study indicate that native isolate of S. carpocapsae cause mortality on the larvae of G. pyloalis and It interferes in the immune and antioxidant responses.","PeriodicalId":14623,"journal":{"name":"ISJ-Invertebrate Survival Journal","volume":null,"pages":null},"PeriodicalIF":0.7,"publicationDate":"2019-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88116849","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-07-04DOI: 10.25431/1824-307X/ISJ.V0I0.105-112
Xiaoxu Zhou, X. Chang, Yaoyao Zhan, Xiuli Wang, Kai Lin
MicroRNAs (miRNAs) are important effectors in mediating host-pathogen interaction. MiR-2005 is observed to be involved in immune response processes in Apostichopus japonicus. In the present study, the putative target genes of miR-2005 in A. japonicus coelomocytes were predicted by bioinformatics analysis of transcriptome database and PCR approaches. A total of 506 potential targets were screened, and 187 targets were annotated. Several immune-related target genes were identified in this study, such as SLI3, CFHR5, FGL, A2ML, and Rab9al. The expression patterns of miR-2005 and its potential targets were validated by quantitative real-time PCR in Vibrio splendidus challenged A. japonicus. For further characterization, an overexpression experiment of miR-2005 at cellular levels was applied. Accordingly, significant negative correlation expression profiles were detected between miR-2005 and two candidates targets, suggesting that SLI3 and CHRP5 showed high possibility to be the targets of miR-2005 in A. japonicus. Altogether, this study will enhance our understanding in the context of miR-2005 modulating the interaction of A. japonicus after being challenged by V. splendidus.
{"title":"Expression analysis of miR-2005 and its target genes in Apostichopus japonicus by Vibrio splendidus challenged","authors":"Xiaoxu Zhou, X. Chang, Yaoyao Zhan, Xiuli Wang, Kai Lin","doi":"10.25431/1824-307X/ISJ.V0I0.105-112","DOIUrl":"https://doi.org/10.25431/1824-307X/ISJ.V0I0.105-112","url":null,"abstract":"MicroRNAs (miRNAs) are important effectors in mediating host-pathogen interaction. MiR-2005 is observed to be involved in immune response processes in Apostichopus japonicus. In the present study, the putative target genes of miR-2005 in A. japonicus coelomocytes were predicted by bioinformatics analysis of transcriptome database and PCR approaches. A total of 506 potential targets were screened, and 187 targets were annotated. Several immune-related target genes were identified in this study, such as SLI3, CFHR5, FGL, A2ML, and Rab9al. The expression patterns of miR-2005 and its potential targets were validated by quantitative real-time PCR in Vibrio splendidus challenged A. japonicus. For further characterization, an overexpression experiment of miR-2005 at cellular levels was applied. Accordingly, significant negative correlation expression profiles were detected between miR-2005 and two candidates targets, suggesting that SLI3 and CHRP5 showed high possibility to be the targets of miR-2005 in A. japonicus. Altogether, this study will enhance our understanding in the context of miR-2005 modulating the interaction of A. japonicus after being challenged by V. splendidus.","PeriodicalId":14623,"journal":{"name":"ISJ-Invertebrate Survival Journal","volume":null,"pages":null},"PeriodicalIF":0.7,"publicationDate":"2019-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78020324","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-07-01DOI: 10.25431/1824-307X/ISJ.V0I0.92-104
F. Cavaliere, P. Brandmayr, P. Giulianini, Maria Luigia Vommaro, A. Giglio
Carabids are of special interest as environmental quality assessment indicators of exposure to xenobiotic and for pest control. In agroecosystems, they can be exposed to a wide range of pathogens and environmental pollution exerting a stronger selection on their innate immune systems. Therefore, information on species-specific immunocompetence is necessary to complete the ecological framework of ground beetles. In this study, cellular and humoral responses were characterized in adults of Harpalus (Pseudoophonus) rufipes (De Geer, 1774) to define a baseline knowledge for future ecotoxicological studies. The circulating hemocytes were characterized by light and transmission electron microscopy and in vivo assay performed by injecting latex beads to identify phagocytizing hemocytes. Ultrastructural analyses revealed four morphologically distinct types of circulating hemocytes: prohemocytes, plasmatocytes, granular cells and oenocytoids. Differential hemocyte counts showed that plasmatocytes and granular cells were the most abundant circulating cell types and granular cells exhibited phagocytic activity following immune challenge with latex beads. Mitotic figures and non-differentiated hemocytes observed under light microscopy indicate a continuous cell turnover in the hemolymph. Melanotic nodules found 2h after the immune challenge were formed to immobilize the latex beads. Phenoloxidase (PO) assays revealed an increase of basal PO activity in hemolymph after immune system activation with lipopolysaccharide (LPS). However, the LPS-stimulated adults showed no significant variation in the lysozyme-like enzyme activity in hemolymph. Based on these results, H. rufipes displays a rapid, non-specific immune response involving cellular and humoral effectors that both sequester and clear pathogens.
瓢虫是特别感兴趣的环境质量评价指标暴露于外源和害虫防治。在农业生态系统中，它们可能暴露于各种各样的病原体和环境污染，对其先天免疫系统产生更强的选择。因此，了解种特异性免疫能力对完善地甲虫的生态框架是必要的。在这项研究中，细胞和体液反应的特征在成年Harpalus (Pseudoophonus) rufipes (De Geer, 1774)，以确定未来的生态毒理学研究的基础知识。通过光学和透射电子显微镜对循环血细胞进行了表征，并通过注射乳胶珠进行了体内检测，以鉴定吞噬血细胞。超微结构分析显示了四种形态不同的循环血细胞:原血细胞、浆细胞、颗粒细胞和嗜酸细胞。不同的血细胞计数显示浆细胞和颗粒细胞是最丰富的循环细胞类型，颗粒细胞在乳胶珠免疫攻击后表现出吞噬活性。光镜下观察到的有丝分裂图和未分化的血细胞表明在血淋巴中有连续的细胞更新。在形成免疫攻击以固定乳胶珠2h后发现黑色结节。用脂多糖(LPS)激活免疫系统后，血淋巴中酚氧化酶(PO)活性增加。而lps刺激的成虫血淋巴溶菌酶样酶活性无明显变化。基于这些结果，猩红嗜血杆菌表现出一种快速的、非特异性的免疫反应，涉及细胞和体液效应，既隔离又清除病原体。
{"title":"Harpalus (Pseudoophonus) rufipes as a model to study cellular and humoral immune defence strategies in coleopteran species","authors":"F. Cavaliere, P. Brandmayr, P. Giulianini, Maria Luigia Vommaro, A. Giglio","doi":"10.25431/1824-307X/ISJ.V0I0.92-104","DOIUrl":"https://doi.org/10.25431/1824-307X/ISJ.V0I0.92-104","url":null,"abstract":"Carabids are of special interest as environmental quality assessment indicators of exposure to xenobiotic and for pest control. In agroecosystems, they can be exposed to a wide range of pathogens and environmental pollution exerting a stronger selection on their innate immune systems. Therefore, information on species-specific immunocompetence is necessary to complete the ecological framework of ground beetles. In this study, cellular and humoral responses were characterized in adults of Harpalus (Pseudoophonus) rufipes (De Geer, 1774) to define a baseline knowledge for future ecotoxicological studies. The circulating hemocytes were characterized by light and transmission electron microscopy and in vivo assay performed by injecting latex beads to identify phagocytizing hemocytes. Ultrastructural analyses revealed four morphologically distinct types of circulating hemocytes: prohemocytes, plasmatocytes, granular cells and oenocytoids. Differential hemocyte counts showed that plasmatocytes and granular cells were the most abundant circulating cell types and granular cells exhibited phagocytic activity following immune challenge with latex beads. Mitotic figures and non-differentiated hemocytes observed under light microscopy indicate a continuous cell turnover in the hemolymph. Melanotic nodules found 2h after the immune challenge were formed to immobilize the latex beads. Phenoloxidase (PO) assays revealed an increase of basal PO activity in hemolymph after immune system activation with lipopolysaccharide (LPS). However, the LPS-stimulated adults showed no significant variation in the lysozyme-like enzyme activity in hemolymph. Based on these results, H. rufipes displays a rapid, non-specific immune response involving cellular and humoral effectors that both sequester and clear pathogens.","PeriodicalId":14623,"journal":{"name":"ISJ-Invertebrate Survival Journal","volume":null,"pages":null},"PeriodicalIF":0.7,"publicationDate":"2019-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83847142","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-05-21DOI: 10.25431/1824-307X/ISJ.V0I0.84-91
R. Simonini, Dario Iori, L. Forti, Sara Righi, D. Prevedelli
Polychaetes play a prominent role in marine systems, but little is known about their secondary metabolites compared with other benthic taxa. In the present study, we investigated the toxicity of hallachrome, an unusual 1-2 anthraquinone identified from the skin of some polychaetes, including the Mediterranean infaunal species Halla parthenopeia. Under stress conditions, this worm releases a harmful purple mucus, whose noxious compounds were still unknown. We hypothesized that hallachrome also occurs in the purple mucus, giving rise to its color and toxicity. Soon after the production of the purple exudate, H. parthenopeia also secretes a harmless, transparent mucus, which pushes away the toxic one, suggesting protective functions for the worm itself. LC-MS and 1H-NMR analyses confirmed the presence of the pigment hallachrome in the purple mucus. The average concentration of the pigment in the purple mucus was about 310 mg L-1. Ecotoxicological bioassays on representative species of bacteria, protozoans, rotifers, crustaceans (Artemia franciscana) and polychaetes (Dinophilus gyrociliatus) revealed its severe toxic effects: LC50/EC50 values ranged from 0.11-5.67 mg L-1. Hallachrome showed higher toxicity for A. franciscana than other naturally occurring anthraquinones. Tests on encapsulated embryos of D. gyrociliatus evidenced the ability of a mucus layer to limit hallachrome diffusion, confirming the protective role of the transparent mucus. Given the information available on polychaetes anti-predator strategies, hallachrome cannot be considered a consumer deterrent. However its toxicity and wide range of activity suggest chemical defensive functions against potential competitors, parasites and/or pathogens.
{"title":"Ecotoxicity of hallachrome, an unusual 1-2 anthraquinone excreted by the infaunal polychaete Halla parthenopeia: evidence for a chemical defence?","authors":"R. Simonini, Dario Iori, L. Forti, Sara Righi, D. Prevedelli","doi":"10.25431/1824-307X/ISJ.V0I0.84-91","DOIUrl":"https://doi.org/10.25431/1824-307X/ISJ.V0I0.84-91","url":null,"abstract":"Polychaetes play a prominent role in marine systems, but little is known about their secondary metabolites compared with other benthic taxa. In the present study, we investigated the toxicity of hallachrome, an unusual 1-2 anthraquinone identified from the skin of some polychaetes, including the Mediterranean infaunal species Halla parthenopeia. Under stress conditions, this worm releases a harmful purple mucus, whose noxious compounds were still unknown. We hypothesized that hallachrome also occurs in the purple mucus, giving rise to its color and toxicity. Soon after the production of the purple exudate, H. parthenopeia also secretes a harmless, transparent mucus, which pushes away the toxic one, suggesting protective functions for the worm itself. LC-MS and 1H-NMR analyses confirmed the presence of the pigment hallachrome in the purple mucus. The average concentration of the pigment in the purple mucus was about 310 mg L-1. Ecotoxicological bioassays on representative species of bacteria, protozoans, rotifers, crustaceans (Artemia franciscana) and polychaetes (Dinophilus gyrociliatus) revealed its severe toxic effects: LC50/EC50 values ranged from 0.11-5.67 mg L-1. Hallachrome showed higher toxicity for A. franciscana than other naturally occurring anthraquinones. Tests on encapsulated embryos of D. gyrociliatus evidenced the ability of a mucus layer to limit hallachrome diffusion, confirming the protective role of the transparent mucus. Given the information available on polychaetes anti-predator strategies, hallachrome cannot be considered a consumer deterrent. However its toxicity and wide range of activity suggest chemical defensive functions against potential competitors, parasites and/or pathogens.","PeriodicalId":14623,"journal":{"name":"ISJ-Invertebrate Survival Journal","volume":null,"pages":null},"PeriodicalIF":0.7,"publicationDate":"2019-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76752939","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-05-14DOI: 10.25431/1824-307X/ISJ.V0I0.72-83
Changhao Gong, Chao Liu, Huan Li, M. Li, Zhaoqun Liu, Weilin Wang, Liyan Wang, L. Song
The Pacific oyster Crassostrea gigas is an important species living in the intertidal zones. It is of great significance to study the mechanism of oysters to adapt air exposure. In the present study, weighted gene co-expression network analysis (WGCNA) with the transcriptome data of gills and adductor muscle was conducted to investigate the metabolic transformation of C. gigas under air exposure. GO enrichment of modules specifically expressed in adductor muscle of oysters exposed to air for five, seven and nine days revealed the phased expression of respiratory chain, protein turnover and lipid metabolism, indicating the conversion of energy metabolism. During air exposure, “respiratory chain” and “ribosome biogenesis” were enriched in the muscle on the fifth day, suggesting that glycogen metabolism was dominant in the early stages of air exposure. On the seventh day, many terms about the regulation of proteolysis were enriched, indicating that carbohydrates were not be able to meet the metabolic needs in the oyster adductor muscle, and proteins began to be degraded for energy supply. The processes related to lipid metabolism were enriched on the ninth day. The extremely high glycogen content of C. gigas allowed it to maintain a basic metabolic activity for a long time with a conservative compensation strategy. GO and KEGG enrichments of the modules sensitive to air exposure in gills were mainly involved in “response to endoplasmic reticulum stress”, “Endoplasmic Reticulum (ER) to Golgi transport vesicle membrane” and “protein processing in endoplasmic reticulum”. It revealed that the mechanism of oyster adapting to air exposure was a complex regulatory network depending on the ER. Hub gene network and PPI network analyses found that some transcription factors containing zinc finger domains regulated the biochemical reactions for stress adaptation, indicating that the ER, as a regulatory element sensitive to external stress, could regulate apoptosis, autophagy and protein degradation in gills of C. gigas under air exposure. These results would provide new insights into the adaptation of C. gigas to air exposure in terms of energy metabolism and homeostasis.
{"title":"The transformation of energy metabolism and Endoplasmic Reticulum stress regulation in Pacific oyster Crassostrea gigas under air exposure","authors":"Changhao Gong, Chao Liu, Huan Li, M. Li, Zhaoqun Liu, Weilin Wang, Liyan Wang, L. Song","doi":"10.25431/1824-307X/ISJ.V0I0.72-83","DOIUrl":"https://doi.org/10.25431/1824-307X/ISJ.V0I0.72-83","url":null,"abstract":"The Pacific oyster Crassostrea gigas is an important species living in the intertidal zones. It is of great significance to study the mechanism of oysters to adapt air exposure. In the present study, weighted gene co-expression network analysis (WGCNA) with the transcriptome data of gills and adductor muscle was conducted to investigate the metabolic transformation of C. gigas under air exposure. GO enrichment of modules specifically expressed in adductor muscle of oysters exposed to air for five, seven and nine days revealed the phased expression of respiratory chain, protein turnover and lipid metabolism, indicating the conversion of energy metabolism. During air exposure, “respiratory chain” and “ribosome biogenesis” were enriched in the muscle on the fifth day, suggesting that glycogen metabolism was dominant in the early stages of air exposure. On the seventh day, many terms about the regulation of proteolysis were enriched, indicating that carbohydrates were not be able to meet the metabolic needs in the oyster adductor muscle, and proteins began to be degraded for energy supply. The processes related to lipid metabolism were enriched on the ninth day. The extremely high glycogen content of C. gigas allowed it to maintain a basic metabolic activity for a long time with a conservative compensation strategy. GO and KEGG enrichments of the modules sensitive to air exposure in gills were mainly involved in “response to endoplasmic reticulum stress”, “Endoplasmic Reticulum (ER) to Golgi transport vesicle membrane” and “protein processing in endoplasmic reticulum”. It revealed that the mechanism of oyster adapting to air exposure was a complex regulatory network depending on the ER. Hub gene network and PPI network analyses found that some transcription factors containing zinc finger domains regulated the biochemical reactions for stress adaptation, indicating that the ER, as a regulatory element sensitive to external stress, could regulate apoptosis, autophagy and protein degradation in gills of C. gigas under air exposure. These results would provide new insights into the adaptation of C. gigas to air exposure in terms of energy metabolism and homeostasis.","PeriodicalId":14623,"journal":{"name":"ISJ-Invertebrate Survival Journal","volume":null,"pages":null},"PeriodicalIF":0.7,"publicationDate":"2019-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87755731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-04-19DOI: 10.25431/1824-307X/ISJ.V0I0.66-71
M. Sułek, Lidiia Vertyporokh, P. Waleczko, I. Wojda
Immune priming is a phenomenon that allows invertebrates, which are devoid of acquired immunity, relying on memory T-cells and antibodies, to achieve better protection after subsequent infection. In this short report, we present new investigations of the immune response of primed Galleria mellonella larvae after infection with Bacillus thuringiensis. We compared two related aspects of immune response – hemolymph coagulation and the activity of phenoloxidase in the primed and non-primed larvae after the injection of the lethal dose of B. thuringiensis. The in vivo assay showed that coagulation of hemolymph in response to the bacterial injection occurred more efficiently in the primed animals in comparison to the non-primed ones. Further, we showed that the activity of phenoloxidase was also higher in the primed, infected larvae. Both parameters of insect immune response may contribute to the increased resistance of primed G. mellonella to further infection with B. thuringiensis.
{"title":"Immune priming of Galleria mellonella larvae with Bacillus thuringiensis affects coagulation and phenoloxidase activity upon subsequent infection","authors":"M. Sułek, Lidiia Vertyporokh, P. Waleczko, I. Wojda","doi":"10.25431/1824-307X/ISJ.V0I0.66-71","DOIUrl":"https://doi.org/10.25431/1824-307X/ISJ.V0I0.66-71","url":null,"abstract":"Immune priming is a phenomenon that allows invertebrates, which are devoid of acquired immunity, relying on memory T-cells and antibodies, to achieve better protection after subsequent infection. In this short report, we present new investigations of the immune response of primed Galleria mellonella larvae after infection with Bacillus thuringiensis. We compared two related aspects of immune response – hemolymph coagulation and the activity of phenoloxidase in the primed and non-primed larvae after the injection of the lethal dose of B. thuringiensis. The in vivo assay showed that coagulation of hemolymph in response to the bacterial injection occurred more efficiently in the primed animals in comparison to the non-primed ones. Further, we showed that the activity of phenoloxidase was also higher in the primed, infected larvae. Both parameters of insect immune response may contribute to the increased resistance of primed G. mellonella to further infection with B. thuringiensis.","PeriodicalId":14623,"journal":{"name":"ISJ-Invertebrate Survival Journal","volume":null,"pages":null},"PeriodicalIF":0.7,"publicationDate":"2019-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85658368","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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