Reeves＇s Pheasant （Syrmaticus reevesii） is a threatened pheasant species endemic to China. The habitat use of territorial male birds was surveyed by the help of live decoys in a core area of Dongzhai NationaI Natur...Reeves＇s Pheasant （Syrmaticus reevesii） is a threatened pheasant species endemic to China. The habitat use of territorial male birds was surveyed by the help of live decoys in a core area of Dongzhai NationaI Nature Reserve. The breeding habitat selection of this pheasant was examined at two scales （115 m and 250 m scale, i.e. 4.15 hm^2 and 19.63 hm^2, respectively）, including the characteristics at distance scale. Investigation was based on line transect, RS and GIS in Dongzhai National Natural Reserve from 2001 to 2003. Moreover, a range of habitat variables were compared between used and control points at each scale, and stepwise logistic regression was applied to select the key scale and the key habitat factors in relation to breeding habitat selection of this bird. Our results stated that the territorial males at Baiyun occurred mostly in mixed forests, followed by fir forests, pine forests, shrubs, and broadleaf forests. The area of conifer forests was the key factor influencing habitat selection of this bird in breeding period at the scales of 115 m and 250 m, and the proximity of farmland was important for habitat selection in breeding seasons. Furthermore, Reeves＇s Pheasants attached great importance to the scale of 115 m. When considering a range of habitat variables at all scales within a multivariate regression, the leading factors having effect on habitat selection in the breeding period were areas of conifer forests at 115 m scale and the distance to farmland. In addition, these above results suggested that strengthening the management of suitable habitat, and optimizing the habitat configuration are important in promoting conservation of this bird. However, it also highlighted the importance of initiating future researches on the conifer forests and their impact on the population of Reeves＇s Pheasants, which would be beneficial to promote the habitat conservation of this pheasant more effectively.展开更多
Both copper/zinc superoxide dismutase （SOD; Cu/Zn-SOD, SOD1） cDNA and manganese SOD （Mn-SOD, SOD2） cDNA were cloned for the first time from the three-keeled pond turtle, Chinemys reevesii, using RT-PCR and RACE me...Both copper/zinc superoxide dismutase （SOD; Cu/Zn-SOD, SOD1） cDNA and manganese SOD （Mn-SOD, SOD2） cDNA were cloned for the first time from the three-keeled pond turtle, Chinemys reevesii, using RT-PCR and RACE methods in this work. The SOD1 cDNA was 749 bp long and consisted of a 32-bp 5＇-untranslated region （UTR）, a 249-bp 3＇-UTR, and a 468-bp open reading frame （ORF） encoding a 155-amino-acid protein with 16.0 kDa predicted molecular mass and 5.95 theoretical isoelectric point （p/）. The SOD2 cDNA was 1687 bp long and comprised 94-bp of 5＇-UTR, 912-bp 3＇-UTR and 681-bp ORF encoding a 226-amino-acid protein with 25.0 kDa predicted molecular mass and 8.83 pI. The deduced amino acid sequence of SOD1 showed relatively high similarity （77.4%-87.1%） and identity （65.4%-74.4%） with the published sequences of SOD1 from other vertebrate species, whereas SOD2 protein shared slightly higher similarity （83.6%-95.6%） and identity （76.1%-88.9%） with other reported vertebrates SOD2s. Phylogenetic analysis revealed that the C. reevesii SOD1 and SOD2 were separately clustered together, and were highly conserved during evolution. Both SOD mRNA expression was detected widely in the brain, liver, muscle, kidney, gut, spleen, lung and heart at variable levels. The highest expression of the two SODs was observed in muscle, and followed in brain, liver, kidney, gut and heart, whereas low transcriptional levels were found in spleen and lung. Meanwhile, high activity of SOD 1 was kept in brain, liver, muscle, kidney and heart, and followed in gut, spleen and lung. The activities of SOD2 in brain, liver, muscle, kidney, gut and heart were significantly higher than those in spleen and lung.展开更多
Temperature-dependent sex determination（TSD） is a type of environmental sex determination in which the sex of the embryos depends on the ambient temperature; however,the molecular mechanisms governing this process r...Temperature-dependent sex determination（TSD） is a type of environmental sex determination in which the sex of the embryos depends on the ambient temperature; however,the molecular mechanisms governing this process remain unknown.Aromatase,encoded by the cyp19a1 gene,which converts androgens into estrogens in animals,was considered to be the key gene for TSD.In this study,the 5＇-flanking region of the cyp19a1 gene in Reeves＇ turtle（Mauremys reevesii） was cloned,and the promoter region was identified using the luciferase reporter assay.Then the eggs of Reeves＇ turtle were incubated at different temperatures（26°C： male-biased temperature; 29°C： non-sex-biased temperature and 32°C： female-biased temperature）.During the thermosensitive period,the adrenal kidney gonad complexes（AKG） were sampled.DNA methylation analysis of the AKG samples showed that the promoter region of the cyp19a1 gene was significantly de-methylated in the female-biased temperature regime（P＆lt;0.01）.Quantitative analysis of the cyp19a1 gene and estrogen by q PCR and Elisa assay showed that the expression level of the cyp19a1 gene and estrogen content were both upregulated significantly at the female-biased temperature（P＆lt;0.01）.These results indicated that the de-methylation response of the cyp19a1 gene to incubation temperature,especially at the female-biased temperature,could lead to temperature-specific expression of aromatase and increased estrogen levels,which may further determine gonadal development in Reeves＇ turtle.These findings provide insights into the genetic mechanisms underlying TSD.展开更多
Objective This study was conducted to develop an method for identification of Trachemys scripta elegans(Ba Xi Gui,巴西龟)shell glue from Chinemys reevesii(Zhong Hua Cao Gui,中华草龟)shell glue.Methods In this research...Objective This study was conducted to develop an method for identification of Trachemys scripta elegans(Ba Xi Gui,巴西龟)shell glue from Chinemys reevesii(Zhong Hua Cao Gui,中华草龟)shell glue.Methods In this research,an ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS)method coupled with Binary Compare tool of UNIFI software was validated to differentiate Trachemys scripta elegans(Ba Xi Gui,巴西龟)shell glue from Chinemys reevesii(Zhong Hua Cao Gui,中华草龟)shell glue.The gelatins were digested enzymatically into peptides using trypsin.The UPLC-QTOF/MS data of the trypsin digested samples was subjected to Binary Compare of UNIFI software in order to screen out the unique peptides.PEAKS■Studio software was utilized to identify the sequence of the marker peptides.Results It was found that m/z 641.3 was the marker peptide in samples of Trachemys scripta elegans(Ba Xi Gui,巴西龟)shell glue,and m/z 649.3 was the marker peptide in samples of Chinemys reevesii(Zhong Hua Cao Gui,中华草龟)shell glue,with possible amino acid sequence of GEAGPSGPAGPTGAR and GESGPSGPAGPTGAR respectively.Conclusions The results will be helpful for the differentiating between these 2 gelatins,and it can be also used for quality control of Tortoise shell glue(Gui Jia Jiao,龟甲胶).展开更多
Juvenile three-keeled pond turtles(Chinemys reevesii) were fed diets supplemented with vitamin C(Vc) at doses of 0(basal diet, Vc0), 100(Vc100), 200(Vc200), 500(Vc500) and 2500(Vc2500) mg/kg diets at 28°C for 4 w...Juvenile three-keeled pond turtles(Chinemys reevesii) were fed diets supplemented with vitamin C(Vc) at doses of 0(basal diet, Vc0), 100(Vc100), 200(Vc200), 500(Vc500) and 2500(Vc2500) mg/kg diets at 28°C for 4 weeks, respectively. Then, the water temperature was gradually reduced to 10°C, and the turtles were induced into hibernation. Liver tissue samples were collected at three time points: start of hibernation(T1), 4 and 6 weeks’ hibernation(T2 and T3). A control group fed with the basal diet was set to parallel the whole treatment process, but reared at 28°C constantly. The results showed that hibernation mildly affected the antioxidant system and the influence varied with hibernating time. Hepatic malondialdehyde content of the Vc100 group was significantly lower than that of the other groups at T1. At T2, hepatic MDA in the groups of Vc500 and Vc2500 decreased significantly, while no clear differences were found among all groups at T3. The activities of antioxidant enzymes showed a positive correlation with dietary Vc dose before hibernation. After hibernation, total antioxidant capability was not affected by Vc. Superoxide dismutase activity became similar in different groups at T2, but decreased in higher Vc groups(≥ 200 mg/kg) at T3. Glutathione peroxidase and glutathione-S-transferase activities decreased significantly with dietary Vc supplementation(≥ 100 mg/kg) at T2, but recovered at T3. The result indicates that under normal rearing condition, low dietary Vc supplementation(< 100 mg/kg) might be beneficial to the antioxidant defense system. The effect of dietary Vc on the antioxidant defense system differed during hibernation.展开更多
A large amount of marine noise pollution from coastal industrial construction and the water transportation industry affects the growth and development of marine life,which is a big issue threatening marine organisms.H...A large amount of marine noise pollution from coastal industrial construction and the water transportation industry affects the growth and development of marine life,which is a big issue threatening marine organisms.However,most studies have focused on vertebrates,such as marine mammals and fishes,and little is known about the effects of noise on invertebrates.Therefore,the impacts of low-frequency noise(100,300,and 500 Hz)on behavioral and physiological responsesof the sea slug(Onchidium reevesii)were investigated.Under laboratory conditions,sea slugs were stimulated with low-frequency noise at 100,300,and 500 Hz for 1 h.Then,hemolymph enzyme activities(glucose,albumin,triglycerides,superoxide dismutase,catalase,and malondialdehyde)were measured,and mRNA expression of the hsp70 gene was detected in hemolymph and the nervous system by quantitative realtime polymerase chain reaction(qRT-PCR)analysis,while expression of the hsp70 protein was analyzed by immunohistochemistry.The results demonstrated that sea slugs were away from the stimulus source under the influence of low-frequency noise compared to the control group.Enzyme activities,as well as hsp70 gene mRNA and protein expression levels,were significantly higher in the noiseexposed groups than those in the control group.Overall,these changes indicate that low-frequency noise caused oxidative stress in sea slugs in vivo,and the oxidative damage gradually increased when the noise frequency was increased from 100 to 500 Hz.展开更多
The intestine serves as the primary digestive and essential mucosal immune organ,harboring diverse microbiota that play crucial roles in digestion,absorption,and host immunity.However,the association between the intes...The intestine serves as the primary digestive and essential mucosal immune organ,harboring diverse microbiota that play crucial roles in digestion,absorption,and host immunity.However,the association between the intestinal microbiota and host growth and immunity remains insufficiently established,particularly in ectothermic animals where microbiota are easily influenced by external environments.Herein,a comparative analysis of histology,gene expression,transcriptome,and microbiome was deployed to systematically investigate the potential relevance of growth traits,immune responses,and gut microbiota between Reeves'turtles with inferior(IGP)or superior growth performance(SGP).Our results revealed that,in comparison to the IGP group,the SGP group exhibited histological intestinal structures that were more conducive to digestion and absorption,thereby aligning with its superior growth traits.Moreover,transcriptomic annotation identified 50 differentially expressed genes(DEGs)associated with growth and 70 DEGs involved in immunity.These DEGs showed significant enrichment in growth-and immunity-related GO terms and KEGG pathways.Further RT-PCR analyses validated the expression profiles of several genes related to growth and immunity across multiple tissues.In addition,analysis of microbial abundance revealed the presence of various sensitive indicator genera,such as Lactobacillus and Brevundimonas,which potentially imply host growth performance and immune response.At the phylum level,a more diverse and stable microbial interaction between intestinal microbiota in the SGP group and aquaculture water was predicted to contribute to superior growth performance and immunity.Furthermore,correlation analyses revealed a potential interplay between indicator genera with DEGs associated with growth and immune responses.Taken together,these findings highlight the intricate interplay of intestinal microbiota,environmental microbiota,as well as growth-and immunity-related genes in shaping host traits,providing valuable insights into promoting healthy culture practices for Reeves’turtles from both molecular and microbiological perspectives.展开更多
The sea slug(Onchidium reevesii)inhabits intertidal environments and face environmental pressures and disease threats.The immune system of mollusks plays a crucial role in their defense against pathogens and maintaini...The sea slug(Onchidium reevesii)inhabits intertidal environments and face environmental pressures and disease threats.The immune system of mollusks plays a crucial role in their defense against pathogens and maintaining homeostasis.The molecular mechanisms underlying its immune defense and response to pathogens remain largely unexplored.This study aimed to isolate and identify the cDNA sequence of tumor necrosis factor receptor 1(TNFR1)in the sea slug.The obtained cDNA sequence was named OrTNFR1.Like other members of the TNF receptor superfamily,OrTNFR1 contains a signal peptide,TNF homologous domain,and death-related domain.qPCR results showed that OrTNFR1 was highly expressed in the hepatopancreas,blood,and intestines of sea slug.This indicates that OrTNFR1 plays a role in the immune-related tissues of sea slug and is consistent with the broad expression of TNFR1 in bivalves.To investigate the inducible expression pattern and downstream effects of OrTNFR1 in response to lipopolysaccharide(LPS)stimulation,we studied the functional role of the OrTNFR1 gene by stimulating with LPS in vivo and suppressing OrTNFR1 expression using siRNA technology.Our findings demonstrate that LPS stimulation rapidly up-regulates the expression of OrTNFR1 in the hepatopancreas,blood,and intestine of the sea slug.Moreover,si-TNFR1 effectively inhibits LPS-induced TNFR1 expression and its downstream genes.Additionally,we discovered that inhibiting OrTNFR1 reduces LPS-induced cell apoptosis and the expression of apoptosis-associated protein Caspase.TNFR1 is involved in the immune defense of the sea slug,and blocking TNFR1 effectively attenuates LPS-induced cell apoptosis through the TNFR1 pathway.This finding suggests that inhibiting TNFR1 in the sea slug may serve as a potential approach to reduce cell apoptosis.As a shell-less living in harsh intertidal environments,TNFR1 likely participates in its innate immunity.Therefore,studying the TNFR1 pathway contributes to understanding the innate immune signaling pathway in the sea slug,further elucidating host defense against pathogens and apoptotic functions.展开更多
We used mitochondrial cytochrome b and ND4 genes and 9 microsatellite loci to determine genetic diversity,population structure,evolutionary history,and migration patterns within the Reeves’butterfly lizard Leiolepis ...We used mitochondrial cytochrome b and ND4 genes and 9 microsatellite loci to determine genetic diversity,population structure,evolutionary history,and migration patterns within the Reeves’butterfly lizard Leiolepis reevesii(Agamidae).Considering molecular-based phylogeographical lineages,we then performed niche equivalency and similarity tests between divergent lineages.Phylogenetic analyses based on mitochondrial DNA(mtDNA)data revealed 2 lineages(A and B)diverging≈0.84 million years ago and,respectively,restricted to the northern and southern portions of the Wuzhishan and Yinggeling mountain ranges.Lineage B contains individuals from southern Hainan;Lineage A includes individuals from all other localities and can be further divided into 3 clusters according to microsatellite data.The null hypothesis that the 2 lineages shared identical niches was rejected in all niche equivalency tests,indicating niche shifts during genetic divergence.Similarity tests provided evidence of niche conservatism,suggesting that the 2 lineages share more characteristics of their niche spaces than randomly expected.The niche similarity and equivalency tests indicated a complex niche pattern in which both lineages share a main portion of their ecological spaces.The climatic niche of Lineage B represented a marginal and specialized fraction of the entire ecological space of the climatic niche of Lineage A,with warmer conditions.Isolation caused by orogenesis and subsequent niche divergence,together with local adaptation,may have led to genetic differentiation and further lineage sorting in L.reevesii.展开更多
During low tide intertidal organisms face severe temperature fluctuations.This situation may be more pronounced in the context of climate change.To investigate the molecular response mechanism of Onchidium reevesii wh...During low tide intertidal organisms face severe temperature fluctuations.This situation may be more pronounced in the context of climate change.To investigate the molecular response mechanism of Onchidium reevesii when stressed by hot and cold temperatures,we cloned its Caspase-3 and Caspase-9 genes and detected their expression characteristics using RT-qPCR.The results showed that the full-length of the cDNA of Caspase-3 consist of 1831 bp,and included an open reading frame(ORF)of 894 bp,a 50UTR of 155 bp and a 30 UTR of 300 bp,and encoding for 297 amino acids.The full-length of the cDNA of Caspase-9 consisted of 2425 bp,and included an ORF of 1373 bp,a 50 UTR of 290 bp,and a 30 UTR of 759 bp,and encoding for 457 amino acids.During the evolution of species the two genes have been greatly conserved.A homology analysis and phylogenetic tree reconstruction revealed that O.reevesii is most closely related to Aplysia californica.The RT-qPCR showed that both genes were expressed in all of the tissues and that expression was highest in the hepatopancreas.It is speculated that these genes play an essential role in the immunity of O.reevesii to certain diseases.Conditions of temperature stress(40�C and 10�C)showed that both heat and cold stresses could induce significant changes in both genes.Compared to the control group,under cold stress conditions the levels of expression of Caspase-3 and Caspase-9 were significantly upregulated(P<0.05)in each experimental group.When exposed to a constant temperature of 10�C,their expression peaked after 1 h(4.32-fold,P<0.05;and 5.58-fold,P<0.05,respectively)and was at its lowest after 12 h(1.93-fold,P<0.05;and 2.35-fold,P<0.05,respectively).Their expression was upregulated after 24 h(2.68-fold,P<0.05;and 4.21-fold,P<0.05,respectively).The expression of both genes showed similar trends.Under heat stress,the expressions of the two genes showed a wave pattern.When exposed to a constant temperature of 40�C,their expression was up-regulated after 1,6,and 24 h,and down-regulated after 3 and 12 h.Except at 24 h,the other experimental groups did not differ significantly(P>0.05)from the control group.When subjected to various stress conditions the two genes displayed different expression patterns,indicating that the species has different strategies for coping with temperature changes.It further indicates that the effect of temperature on the species may play a role through the Caspase-dependent pathway.These may help to better understand the response,at the molecular level,of O.reevesii to conditions of hot and cold stress.展开更多
基金supported by the National Sciences Foundation of China (30800103)the National Key Project of Scientific and Technical Supporting Programs Funded by Ministry of Science & Technology of China (2008BADB0B01)~~
文摘Reeves＇s Pheasant （Syrmaticus reevesii） is a threatened pheasant species endemic to China. The habitat use of territorial male birds was surveyed by the help of live decoys in a core area of Dongzhai NationaI Nature Reserve. The breeding habitat selection of this pheasant was examined at two scales （115 m and 250 m scale, i.e. 4.15 hm^2 and 19.63 hm^2, respectively）, including the characteristics at distance scale. Investigation was based on line transect, RS and GIS in Dongzhai National Natural Reserve from 2001 to 2003. Moreover, a range of habitat variables were compared between used and control points at each scale, and stepwise logistic regression was applied to select the key scale and the key habitat factors in relation to breeding habitat selection of this bird. Our results stated that the territorial males at Baiyun occurred mostly in mixed forests, followed by fir forests, pine forests, shrubs, and broadleaf forests. The area of conifer forests was the key factor influencing habitat selection of this bird in breeding period at the scales of 115 m and 250 m, and the proximity of farmland was important for habitat selection in breeding seasons. Furthermore, Reeves＇s Pheasants attached great importance to the scale of 115 m. When considering a range of habitat variables at all scales within a multivariate regression, the leading factors having effect on habitat selection in the breeding period were areas of conifer forests at 115 m scale and the distance to farmland. In addition, these above results suggested that strengthening the management of suitable habitat, and optimizing the habitat configuration are important in promoting conservation of this bird. However, it also highlighted the importance of initiating future researches on the conifer forests and their impact on the population of Reeves＇s Pheasants, which would be beneficial to promote the habitat conservation of this pheasant more effectively.
基金supported by a grant from the National Natural Science Foundation of China (Project No. 31172383)
文摘Both copper/zinc superoxide dismutase （SOD; Cu/Zn-SOD, SOD1） cDNA and manganese SOD （Mn-SOD, SOD2） cDNA were cloned for the first time from the three-keeled pond turtle, Chinemys reevesii, using RT-PCR and RACE methods in this work. The SOD1 cDNA was 749 bp long and consisted of a 32-bp 5＇-untranslated region （UTR）, a 249-bp 3＇-UTR, and a 468-bp open reading frame （ORF） encoding a 155-amino-acid protein with 16.0 kDa predicted molecular mass and 5.95 theoretical isoelectric point （p/）. The SOD2 cDNA was 1687 bp long and comprised 94-bp of 5＇-UTR, 912-bp 3＇-UTR and 681-bp ORF encoding a 226-amino-acid protein with 25.0 kDa predicted molecular mass and 8.83 pI. The deduced amino acid sequence of SOD1 showed relatively high similarity （77.4%-87.1%） and identity （65.4%-74.4%） with the published sequences of SOD1 from other vertebrate species, whereas SOD2 protein shared slightly higher similarity （83.6%-95.6%） and identity （76.1%-88.9%） with other reported vertebrates SOD2s. Phylogenetic analysis revealed that the C. reevesii SOD1 and SOD2 were separately clustered together, and were highly conserved during evolution. Both SOD mRNA expression was detected widely in the brain, liver, muscle, kidney, gut, spleen, lung and heart at variable levels. The highest expression of the two SODs was observed in muscle, and followed in brain, liver, kidney, gut and heart, whereas low transcriptional levels were found in spleen and lung. Meanwhile, high activity of SOD 1 was kept in brain, liver, muscle, kidney and heart, and followed in gut, spleen and lung. The activities of SOD2 in brain, liver, muscle, kidney, gut and heart were significantly higher than those in spleen and lung.
基金supported financially by the National Natural Science Foundation of China(Nos.31401053 and 31471966)Guangdong Provincial Natural Science Foundation of China(No.2015A030313903)+1 种基金GDAS Special Project of Science and Technology Development(2017GDASCX-0107)the Funds for Environment Construction and Capacity Building of GDAS’Research Platform(2016GDASPT-0107)
文摘Temperature-dependent sex determination（TSD） is a type of environmental sex determination in which the sex of the embryos depends on the ambient temperature; however,the molecular mechanisms governing this process remain unknown.Aromatase,encoded by the cyp19a1 gene,which converts androgens into estrogens in animals,was considered to be the key gene for TSD.In this study,the 5＇-flanking region of the cyp19a1 gene in Reeves＇ turtle（Mauremys reevesii） was cloned,and the promoter region was identified using the luciferase reporter assay.Then the eggs of Reeves＇ turtle were incubated at different temperatures（26°C： male-biased temperature; 29°C： non-sex-biased temperature and 32°C： female-biased temperature）.During the thermosensitive period,the adrenal kidney gonad complexes（AKG） were sampled.DNA methylation analysis of the AKG samples showed that the promoter region of the cyp19a1 gene was significantly de-methylated in the female-biased temperature regime（P＆lt;0.01）.Quantitative analysis of the cyp19a1 gene and estrogen by q PCR and Elisa assay showed that the expression level of the cyp19a1 gene and estrogen content were both upregulated significantly at the female-biased temperature（P＆lt;0.01）.These results indicated that the de-methylation response of the cyp19a1 gene to incubation temperature,especially at the female-biased temperature,could lead to temperature-specific expression of aromatase and increased estrogen levels,which may further determine gonadal development in Reeves＇ turtle.These findings provide insights into the genetic mechanisms underlying TSD.
基金the funding support from the Aid Program for Science and Technology Innovative Research Team in Higher Educational Institutions of Hunan Province([2010]212)Changsha City Science and Technology Project in 2015(k1501024-31)+3 种基金Key Discipline Program of State Administration of Traditional Chinese Medicine of the P.R.C([2009]30)Key Discipline in Hunan Province([2011]76)Provincial Natural Science Foundation of Hunan(2017JJ3234)Key Project of Science and Technology of Hunan Province(2016SK2033)
文摘Objective This study was conducted to develop an method for identification of Trachemys scripta elegans(Ba Xi Gui,巴西龟)shell glue from Chinemys reevesii(Zhong Hua Cao Gui,中华草龟)shell glue.Methods In this research,an ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS)method coupled with Binary Compare tool of UNIFI software was validated to differentiate Trachemys scripta elegans(Ba Xi Gui,巴西龟)shell glue from Chinemys reevesii(Zhong Hua Cao Gui,中华草龟)shell glue.The gelatins were digested enzymatically into peptides using trypsin.The UPLC-QTOF/MS data of the trypsin digested samples was subjected to Binary Compare of UNIFI software in order to screen out the unique peptides.PEAKS■Studio software was utilized to identify the sequence of the marker peptides.Results It was found that m/z 641.3 was the marker peptide in samples of Trachemys scripta elegans(Ba Xi Gui,巴西龟)shell glue,and m/z 649.3 was the marker peptide in samples of Chinemys reevesii(Zhong Hua Cao Gui,中华草龟)shell glue,with possible amino acid sequence of GEAGPSGPAGPTGAR and GESGPSGPAGPTGAR respectively.Conclusions The results will be helpful for the differentiating between these 2 gelatins,and it can be also used for quality control of Tortoise shell glue(Gui Jia Jiao,龟甲胶).
文摘Juvenile three-keeled pond turtles(Chinemys reevesii) were fed diets supplemented with vitamin C(Vc) at doses of 0(basal diet, Vc0), 100(Vc100), 200(Vc200), 500(Vc500) and 2500(Vc2500) mg/kg diets at 28°C for 4 weeks, respectively. Then, the water temperature was gradually reduced to 10°C, and the turtles were induced into hibernation. Liver tissue samples were collected at three time points: start of hibernation(T1), 4 and 6 weeks’ hibernation(T2 and T3). A control group fed with the basal diet was set to parallel the whole treatment process, but reared at 28°C constantly. The results showed that hibernation mildly affected the antioxidant system and the influence varied with hibernating time. Hepatic malondialdehyde content of the Vc100 group was significantly lower than that of the other groups at T1. At T2, hepatic MDA in the groups of Vc500 and Vc2500 decreased significantly, while no clear differences were found among all groups at T3. The activities of antioxidant enzymes showed a positive correlation with dietary Vc dose before hibernation. After hibernation, total antioxidant capability was not affected by Vc. Superoxide dismutase activity became similar in different groups at T2, but decreased in higher Vc groups(≥ 200 mg/kg) at T3. Glutathione peroxidase and glutathione-S-transferase activities decreased significantly with dietary Vc supplementation(≥ 100 mg/kg) at T2, but recovered at T3. The result indicates that under normal rearing condition, low dietary Vc supplementation(< 100 mg/kg) might be beneficial to the antioxidant defense system. The effect of dietary Vc on the antioxidant defense system differed during hibernation.
基金supported by the Shanghai Collaborative Innovation Center for Cultivating Elite Breeds and Green-culture of Aquaculture Animals(No.A1-3605-21-000202)the Capacity Enhancement of Aquatic Germplasm Resources Research and Support Platform of Shanghai Ocean University(No.A1-3201-20-300206).
文摘A large amount of marine noise pollution from coastal industrial construction and the water transportation industry affects the growth and development of marine life,which is a big issue threatening marine organisms.However,most studies have focused on vertebrates,such as marine mammals and fishes,and little is known about the effects of noise on invertebrates.Therefore,the impacts of low-frequency noise(100,300,and 500 Hz)on behavioral and physiological responsesof the sea slug(Onchidium reevesii)were investigated.Under laboratory conditions,sea slugs were stimulated with low-frequency noise at 100,300,and 500 Hz for 1 h.Then,hemolymph enzyme activities(glucose,albumin,triglycerides,superoxide dismutase,catalase,and malondialdehyde)were measured,and mRNA expression of the hsp70 gene was detected in hemolymph and the nervous system by quantitative realtime polymerase chain reaction(qRT-PCR)analysis,while expression of the hsp70 protein was analyzed by immunohistochemistry.The results demonstrated that sea slugs were away from the stimulus source under the influence of low-frequency noise compared to the control group.Enzyme activities,as well as hsp70 gene mRNA and protein expression levels,were significantly higher in the noiseexposed groups than those in the control group.Overall,these changes indicate that low-frequency noise caused oxidative stress in sea slugs in vivo,and the oxidative damage gradually increased when the noise frequency was increased from 100 to 500 Hz.
基金conducted by supporting from KoreaNational Park Service(KNPS)National Institute of Biological Resource(NIBR) Korea Environmental Industry&Technology Institute(KEITI)
基金financially supported by National Natural Science Foundation of China(32173021)Natural Science Foundation of Hunan Province(2024JJ5184)+3 种基金Hunan Provincial Science&Technology Major Project(2022sfq30,20231F18)Key Projects of Hunan Provincial Education Department(23A0196)Science and Technology Innovation Program of Hunan Province(2024RC8250)Earmarked Fund for Hunan Agriculture Research System(HARS-07).
文摘The intestine serves as the primary digestive and essential mucosal immune organ,harboring diverse microbiota that play crucial roles in digestion,absorption,and host immunity.However,the association between the intestinal microbiota and host growth and immunity remains insufficiently established,particularly in ectothermic animals where microbiota are easily influenced by external environments.Herein,a comparative analysis of histology,gene expression,transcriptome,and microbiome was deployed to systematically investigate the potential relevance of growth traits,immune responses,and gut microbiota between Reeves'turtles with inferior(IGP)or superior growth performance(SGP).Our results revealed that,in comparison to the IGP group,the SGP group exhibited histological intestinal structures that were more conducive to digestion and absorption,thereby aligning with its superior growth traits.Moreover,transcriptomic annotation identified 50 differentially expressed genes(DEGs)associated with growth and 70 DEGs involved in immunity.These DEGs showed significant enrichment in growth-and immunity-related GO terms and KEGG pathways.Further RT-PCR analyses validated the expression profiles of several genes related to growth and immunity across multiple tissues.In addition,analysis of microbial abundance revealed the presence of various sensitive indicator genera,such as Lactobacillus and Brevundimonas,which potentially imply host growth performance and immune response.At the phylum level,a more diverse and stable microbial interaction between intestinal microbiota in the SGP group and aquaculture water was predicted to contribute to superior growth performance and immunity.Furthermore,correlation analyses revealed a potential interplay between indicator genera with DEGs associated with growth and immune responses.Taken together,these findings highlight the intricate interplay of intestinal microbiota,environmental microbiota,as well as growth-and immunity-related genes in shaping host traits,providing valuable insights into promoting healthy culture practices for Reeves’turtles from both molecular and microbiological perspectives.
基金Shanghai Collaborative Innovation Center for Cultivating Elite Breeds and Green-culture of Aquaculture animals(A1-3605-21-000202)the Capacity Enhancement of Aquatic Germplasm Resources Research and Support Platform of Shanghai Ocean University(A1-3201-20-300206).
文摘The sea slug(Onchidium reevesii)inhabits intertidal environments and face environmental pressures and disease threats.The immune system of mollusks plays a crucial role in their defense against pathogens and maintaining homeostasis.The molecular mechanisms underlying its immune defense and response to pathogens remain largely unexplored.This study aimed to isolate and identify the cDNA sequence of tumor necrosis factor receptor 1(TNFR1)in the sea slug.The obtained cDNA sequence was named OrTNFR1.Like other members of the TNF receptor superfamily,OrTNFR1 contains a signal peptide,TNF homologous domain,and death-related domain.qPCR results showed that OrTNFR1 was highly expressed in the hepatopancreas,blood,and intestines of sea slug.This indicates that OrTNFR1 plays a role in the immune-related tissues of sea slug and is consistent with the broad expression of TNFR1 in bivalves.To investigate the inducible expression pattern and downstream effects of OrTNFR1 in response to lipopolysaccharide(LPS)stimulation,we studied the functional role of the OrTNFR1 gene by stimulating with LPS in vivo and suppressing OrTNFR1 expression using siRNA technology.Our findings demonstrate that LPS stimulation rapidly up-regulates the expression of OrTNFR1 in the hepatopancreas,blood,and intestine of the sea slug.Moreover,si-TNFR1 effectively inhibits LPS-induced TNFR1 expression and its downstream genes.Additionally,we discovered that inhibiting OrTNFR1 reduces LPS-induced cell apoptosis and the expression of apoptosis-associated protein Caspase.TNFR1 is involved in the immune defense of the sea slug,and blocking TNFR1 effectively attenuates LPS-induced cell apoptosis through the TNFR1 pathway.This finding suggests that inhibiting TNFR1 in the sea slug may serve as a potential approach to reduce cell apoptosis.As a shell-less living in harsh intertidal environments,TNFR1 likely participates in its innate immunity.Therefore,studying the TNFR1 pathway contributes to understanding the innate immune signaling pathway in the sea slug,further elucidating host defense against pathogens and apoptotic functions.
基金This work was supported by the National Natural Science Foundation of China(31971414 and 31470471)the Finance Science and Technology Project of Hainan Province(ZDYF2018219)。
文摘We used mitochondrial cytochrome b and ND4 genes and 9 microsatellite loci to determine genetic diversity,population structure,evolutionary history,and migration patterns within the Reeves’butterfly lizard Leiolepis reevesii(Agamidae).Considering molecular-based phylogeographical lineages,we then performed niche equivalency and similarity tests between divergent lineages.Phylogenetic analyses based on mitochondrial DNA(mtDNA)data revealed 2 lineages(A and B)diverging≈0.84 million years ago and,respectively,restricted to the northern and southern portions of the Wuzhishan and Yinggeling mountain ranges.Lineage B contains individuals from southern Hainan;Lineage A includes individuals from all other localities and can be further divided into 3 clusters according to microsatellite data.The null hypothesis that the 2 lineages shared identical niches was rejected in all niche equivalency tests,indicating niche shifts during genetic divergence.Similarity tests provided evidence of niche conservatism,suggesting that the 2 lineages share more characteristics of their niche spaces than randomly expected.The niche similarity and equivalency tests indicated a complex niche pattern in which both lineages share a main portion of their ecological spaces.The climatic niche of Lineage B represented a marginal and specialized fraction of the entire ecological space of the climatic niche of Lineage A,with warmer conditions.Isolation caused by orogenesis and subsequent niche divergence,together with local adaptation,may have led to genetic differentiation and further lineage sorting in L.reevesii.
基金This work was supported by the Construction Project of the Double First-class Disciplines of Fisheries.
文摘During low tide intertidal organisms face severe temperature fluctuations.This situation may be more pronounced in the context of climate change.To investigate the molecular response mechanism of Onchidium reevesii when stressed by hot and cold temperatures,we cloned its Caspase-3 and Caspase-9 genes and detected their expression characteristics using RT-qPCR.The results showed that the full-length of the cDNA of Caspase-3 consist of 1831 bp,and included an open reading frame(ORF)of 894 bp,a 50UTR of 155 bp and a 30 UTR of 300 bp,and encoding for 297 amino acids.The full-length of the cDNA of Caspase-9 consisted of 2425 bp,and included an ORF of 1373 bp,a 50 UTR of 290 bp,and a 30 UTR of 759 bp,and encoding for 457 amino acids.During the evolution of species the two genes have been greatly conserved.A homology analysis and phylogenetic tree reconstruction revealed that O.reevesii is most closely related to Aplysia californica.The RT-qPCR showed that both genes were expressed in all of the tissues and that expression was highest in the hepatopancreas.It is speculated that these genes play an essential role in the immunity of O.reevesii to certain diseases.Conditions of temperature stress(40�C and 10�C)showed that both heat and cold stresses could induce significant changes in both genes.Compared to the control group,under cold stress conditions the levels of expression of Caspase-3 and Caspase-9 were significantly upregulated(P<0.05)in each experimental group.When exposed to a constant temperature of 10�C,their expression peaked after 1 h(4.32-fold,P<0.05;and 5.58-fold,P<0.05,respectively)and was at its lowest after 12 h(1.93-fold,P<0.05;and 2.35-fold,P<0.05,respectively).Their expression was upregulated after 24 h(2.68-fold,P<0.05;and 4.21-fold,P<0.05,respectively).The expression of both genes showed similar trends.Under heat stress,the expressions of the two genes showed a wave pattern.When exposed to a constant temperature of 40�C,their expression was up-regulated after 1,6,and 24 h,and down-regulated after 3 and 12 h.Except at 24 h,the other experimental groups did not differ significantly(P>0.05)from the control group.When subjected to various stress conditions the two genes displayed different expression patterns,indicating that the species has different strategies for coping with temperature changes.It further indicates that the effect of temperature on the species may play a role through the Caspase-dependent pathway.These may help to better understand the response,at the molecular level,of O.reevesii to conditions of hot and cold stress.
