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Information
月刊，1984年创刊
主 管：内蒙古自治区教育厅
主 办：包头医学院
主 编：赵云山
特邀主编：高长青
编辑出版：包头医学院学报编辑部
国际刊号：ISSN 1006－740X
国内刊号：CN 15－1182/R
邮发代号：16-292
Links
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2023 Vol. 42, No. 4
Published: 2023-07-25
Research and Application
Overview and Specific Topi
Research and Application
547
Identification of Reference Genes in Adipose Tissue and Adipocytes of Grass Carp
Ctenopharyngodon idellus
by qRT-PCR
HU Zechao, ZOU Xiaocui, SUN Jian, BIAN Chenchen, JI Hong
DOI: 10.16378/j.cnki.1003-1111.21102
To explore the stable expression of the four reference genes in grass carp
Ctenopharyngodon idellus
adipose tissue and adipocytes, and to select the best stable reference genes, transcription levels of 4 candidate reference genes including 18S rRNA, β-actin, GAPDH and EF1α were investigated in abdominal adipose tissue of grass carp with body weight of (15.0±0.5) g fed diets containing 4%, 8% and 12% of lipid at water temperature of 26.5—27.5 ℃ for 4 weeks when the adipocytes were treated with oleic acid concentrations of 0, 40, 80 and 120 mmol/L by real-time fluorescence quantitative PCR (qRT-PCR), and then the stability of their expression levels were analyzed by software GeNorm, NormFinder and BestKeeper to screen out the reference genes or combinations stably expressed in adipose tissue and adipose cells. The results showed that specific amplification products and ideal amplification efficiency were obtained by all the quantitative PCR primers for candidate reference genes. The descending order of expression stability for candidate reference genes was expressed as 18S rRNA>β-actin>GAPDH>EF1α and 18S rRNA=β-actin>GAPDH>EF1αIn in adipose tissue and adipocytes. The differentiation analysis of standardized factor pairing of GeNorm showed that its
V
n
/
V
(n +1)
was found to be less than 0.15, that is, with the optimal reference genes number of at least 2 in adipose tissue and adipocytes. The findings indicated that the reference gene stably expressed in adipose tissue and adipocytes of grass carp was 18S rRNA. It was suggested that 18S rRNA and β-actin should be selected as the most suitable reference gene combination in adipose tissue and cells.
2023 Vol. 42 (4): 547-555 [
Abstract
] (
226
)
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556
Effects of Different Straw-Returning on Macrobenthic Community Structure in Rice-Crayfish Fields During Winter and Spring
SUN Ying, LI Wei, XI Yewen, CHENG Yongxu, LI Jiayao
DOI: 10.16378/j.cnki.1003-1111.21063
The purpose of this experiment is to study the characteristics of macrobenthic community changes before and after the application of red swamp crayfish
Procambarus clarkia
under the treatment of the amount of straw-returning to the rice field in winter and spring. Macrobenthic community structure including diversity, density and biomass were analyzed in red swamp crayfish in several 125 m
2
ricecrayfish fields at Shanghai Ocean University Chongming Base (N 31°34', E 121°33') with varying amounts of straw-returning including 0(CK), 1500 kg/hm
2
(S1), 3500 kg/hm
2
(S2), and 4500 kg/hm
2
(S3) from December 2018 to July 2019. The results showed that before and after releasing crayfish, diversity index is presented as S2>S3>S1>CK and CK> S2> S3>S1,respectively. Before the release of crayfish, the relationship between total density, biomass and the amount of returning all showed as S3>S2>S1>CK. After the release of crayfish, the total density of S1, S2 and S3 treatment was dropped significantly. The calculation of the dominance index found that the density of
Branchiura sowerbyi
,
Limnodrilus hoffmeisteri
,
Hippeutis umbilicalis
Benson was decreased significantly (
P
<0.05), suggesting that these might be important natural food for crayfish. Among them, the density of dominant species in S2 treatment was dropped significantly higher than that in S1 and S3 treatments. Redundancy analysis (RDA) showed that soil total phosphorus, soil bulk density and organic matter were the key factors influencing the macrobenthic community structure. Under the experimental conditions, considering the changes of macrobenthos community structure, soil fertility and aquaculture water indicators before and after crayfish feeding, S2 is the most suitable amount of straw returning to the field.
2023 Vol. 42 (4): 556-565 [
Abstract
] (
141
)
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566
Relationship between Morphological Traits and Body Weight of Chinese Shrimp
Fenneropenaeus chinensis
of Different Genders
ZHENG Li, HE Yuying, WANG Qiong, YU An, XIE Yongjun
DOI: 10.16378/j.cnki.1003-1111.21124
To study the influence of morphological traits on body weight of Chinese shrimp
Fenneropenaeus chinensis
of different genders, 158 males and 158 females were selected. The body length (
x
1
), frontal length (
x
2
), carapace length (
x
3
), carapace width (
x
4
), abdominal section long (
x
5
), tail length (
x
6
), external flagellum length of first antenna (
x
7
), internal flagellum length of first antenna (
x
8
), second antenna length (
x
9
), number of above rostral teeth (
x
10
), number of below rostral teeth (
x
11
) and body weight (
y
) were measured, and multiple regression analysis and path analysis were used to analyze the effects of various morphological traits on body weight. The results showed that the correlation between body length and body weight was the largest in both male and female populations, which were 0.885 and 0.869, respectively. In female populations, except the number of above rostral teeth, other morphological traits were significantly or extremely significantly positively correlated with body weight. Except for the number of above rostral teeth and below rostral teeth, other morphological traits of male population were positively correlated with body weight.The morphological traits that had the greatest direct effect on body weight were body length, with path coefficients of 0.401 and 0.673, respectively. In terms of indirect effects, body length and abdominal section long had greater indirect effects on body weight through other morphological traits, which were 1.897 and 1.528, respectively. In the male populations, body length has the largest indirect effect on body weight through other morphological traits, which is 2.159. Body length was the common morphological trait that determined the body weight of the male and female populations. In addition, the external flagellum length of first antenna, the abdominal section long and the number of below rostral teeth had a greater effect on the body weight of the female population, while the carapace length and the second antenna length had a greater effect on the body weight of the male population. Using morphological traits as independent variables and body weight as dependent variables, the regression equations of different sexes of Chinese shrimp were established. The values of male and female were
y
1
=-82.999+3.644
x
1
+4.992
x
5
+1.208
x
7
+2.126
x
11
(
r
2
=0.927),
y
2
=-46.347+3.757
x
1
+1.189
x
3
+0.143
x
9
(
r
2
=0.896)
2023 Vol. 42 (4): 566-574 [
Abstract
] (
160
)
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126
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575
DNA Barcode for Species Identification and Phylogenetic Relationship of Bagridae Fish
LIU Hongyan, XIONG Fei, ZHAI Dongdong, WANG Ying, XIA Ming, CHEN Yuanyuan
DOI: 10.16378/j.cnki.1003-1111.21089
The family Bagridae is a diverse fish group, some species with highly similar traits are difficult to be identified and the phylogenetic relationship are ambiguity. Therefore, an accurate and convenient method for identifying and classifying is important and necessary. Herein, we provided a molecular method based on mitochondrial cytochrome c subunit Ⅰ (COⅠ) for the identification and analysis of phylogenetic relationship of Bagridae fish. Two hundred and fourteen mitochondrial COⅠ gene sequences belonging to thirty-six species of five genera were studied. The average genetic distances of intraspecific and interspecific species were 0.016 and 0.158, respectively. The average interspecific genetic distance was 9.875 times of the average intraspecific genetic distance. It existed barcode gaps between interspecific distance and intraspecific distance. ABGD analysis showed that 36 species were divided into 25 operational taxonomic units (OTU), The sorting of OTU were roughly consistent with the genetic distance method. The interspecific genetic distance of several species forming one OTU were small, and the intraspecific genetic distance of one species separating into several OTUs were large. The members of
Pelteobagrus
,
Pseudobagrus
and
Leiocassis
mixed together and clustered into one large branch. There was one species each clustering into opposite genus in the genus of
Mystus
and
Hemibagrus,
which indicated that it existed incomplete lineage sorting in the two genera. The results indicated that most of Bagridae species could be identified by DNA barcode, but there were limitations in barcode identification of some closely related species. In addition, DNA barcoding had clarified the evolutionary relationships and would provide a scientific reference for phylogenetic classification of Bagridae fish.
2023 Vol. 42 (4): 575-584 [
Abstract
] (
140
)
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61
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585
Characterization and Expression Analyses of Prostaglandin E
2
Receptor EP4 Gene in
Procambarus clarkii
JIANG Hongxia, ZHANG Ran, LI Yizheng, LIU Xuewei, ZHANG Shuaishuai, YU Miao, WANG Lei, ZHANG Meng, QIAO Zhigang, LI Xuejun
DOI: 10.16378/j.cnki.1003-1111.21033
Prostaglandin E
2
(PGE
2
) receptor EP4 plays an important role in the regulation of ovarian development, ovulation and reproduction in animals. The cDNA sequence of the prostaglandin E
2
(PGE
2
) receptor EP4 gene with the complete ORF region was obtained from the ovary of
Procambarus clarkii
using transcriptome sequencing. In order to explore the sequence characteristics and the function of EP4 gene in ovary of
P. clarkii
(PcEP4), the gene sequence was analyzed by bioinformatics, and the gene expression pattern was detected by real-time fluorescent quantitative PCR. Bioinformatics analysis results showed that the ORF of PcEP4 was 2505 bp, encoding a deduced protein with 835 amino acids; PcEP4 is a typical G protein coupled receptor with 7 transmembrane α-helical structures; EP4 amino acid sequence of
P. clarkii
and
Litopenaeus vannamei
were the most consistent,and the genetic relationship was the closest. Gene expression analysis results showed that PcEP4 gene expression level was the highest in the ovary in
P. clarkii
; in the Ⅰ—Ⅵ stage of ovaries in
P. clarkii
, PcEP4 gene expression level was the highest in stage Ⅴ ovary; At 36, 48 and 60 h after PGE
2
injection, the expression level of PcEP4 gene in the ovaries of
P. clarkii
was significantly higher than that in the control group (
P
<0.05), the egg holding rate of
P. clarkii
also increased compared with the control group at 72 h after PGE
2
injection. The above results indicate that, as a PGE
2
receptor, EP4 plays an important role in promoting oocyte maturation and ovulation of
P. clarkii
together with PGE
2
.
2023 Vol. 42 (4): 585-593 [
Abstract
] (
102
)
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594
Development of Larva, Irreversible Starvation Point and Optimal Stocking Time of
Phoxinus lagowskii
Larva
REN Xingyue, LUO Xiaonian, DENG Shuping, DUAN Youjian, JIANG Yusheng, LI Jiao
DOI: 10.16378/j.cnki.1003-1111.21247
In order to study the developmental biological characteristics of
Phoxinus lagowskii
larva and optimal stocking time, microscopic observation of the
P. lagowskii
larval development, the larval starvation test and larva pond experiment were carried out. The observation of postembryonic development of larva showed that: under the condition of water temperature 16—18 ℃, at the beginning of their
P. lagowskii
length was (4.69±0.37) mm, yolk sac total length was (3.12±0.10) mm, 2 days post hatching (dph) the color of
P. lagowskii
eyespots became black, the digestive tract began versed at 5 dph, and the silver fish swim bladder began inflating one room at 6 dph, when the larval notochord began to bend and initial feeding was observed in the larvae at 7 dph, the yolk sac was completely exhausted at 9 dph, and the larvae showed second chamber of swim bladder at 12 dph, second bladder chamber aeration period at 21 dph, the larvae dorsal and anal fin emerged at 23 dph, at 27 dph larval ventral fins began to form, until at 35 dph, all fins were well developed in the larvae. The experiment of
P. lagowskii
irreversible starvation point results showed that: under 16—18 ℃, the
P. lagowskii
larvae first day of open mouth (at 7 dph) feeding rate was 36.36%. At 9 dph, the highest initial feeding rate was 85.71%, the irreversible starvation point was at 14 dph, the larvae of feeding rate was 40.91%. length (
L
) and age (
t
) von-bertalanffy growth formulaof juvenile fish before mouth open was
L
=8.0031[1-e
-0.3139(
t
+2.8093)
]. The optimal stocking time of
P. lagowskii
results showed that water temperature was 14.5—20.1 ℃, rotifer biomass keeps 8.30—18.58 mg/L, the highest survival rate and time of body mass growth appeared at 6 dph, the survival rate was (75.0±2.2)% and the larvae body mass reached (9.0±2.0) mg. Under the optimum total length of largest growth of timely for 8 dph, the minimum total length of the coefficient of variation was at 7.5 dph, minimum weight coefficient of variation of timely under the pond at was 8.5 dph. This experiment provides basic materials for post-embryonic development and optimal stocking time in
P. lagowskii
larva.
2023 Vol. 42 (4): 594-603 [
Abstract
] (
84
)
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67
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604
Spatial and Temporal Distribution of the Fishing Ground Gravity of
Thunnus alalunga
in Western and Central Pacific
XU Zhenqi, WANG Jintao, LEI Lin, CHEN Xinjun
DOI: 10.16378/j.cnki.1003-1111.21138
As a highly migratory oceanic fish,
Thunnus alalunga
has high economic value. The research and analysis of the spatial and temporal distribution of its resources are the basis for further efficient development and utilization and improvement of operational efficiency. This paper was based on the world longline fishing data of
Thunnus alalunga
in the Western and Central Pacific Ocean from 2009 to 2018. Combined with the data, the monthly variation trend and spatial distribution characteristics of
Thunnus alalunga
resources were explored by using statistics and K-means clustering method. The results showed that the center of gravity of the fishing ground is mainly distributed in the range of E 155°—W 160° and S 15°—S 30° during 2009—2018. The Northern Hemisphere winter (from December to February of the next year) is mainly concentrated in the low latitude area of the northern hemisphere (N 0°—N 10°), the center of gravity of fishing ground shifts to the northwest, and the southern hemisphere winter (from June to August) is mainly concentrated in the low latitude area of the southern hemisphere (S 12°—S 22.5°), the center of gravity of the fishing ground shifted to the southeast. The spatial and temporal distribution of the center of gravity of the fishing ground is significantly affected by the sea surface temperature anomaly. When the SSTA is higher, the center of gravity of the fishing ground is scattered in the central sea area, and when the SSTA is relatively lower, the center of gravity of the fishing ground shifts to the southeast and northwest. The results are helpful to the sustainable development, utilization and scientific management of
Thunnus alalunga
in this sea area.
2023 Vol. 42 (4): 604-612 [
Abstract
] (
135
)
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(1 KB)
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(13618 KB) (
35
)
613
Cloning and Expression Analysis of Byssal Protein 3 Gene in Pearl Oyster
Pteria penguin
YANG Lei, CHEN Yi, ZHANG Jiayi, ZHAN Xin
DOI: 10.16378/j.cnki.1003-1111.21120
In order to understand the molecular mechanism of byssus adhesion of the winged pearl oyster
Pteria penguin
, the full-length cDNA sequence of the byssal protein 3 (BP3) gene was obtained by the rapid amplification of cDNA ends (RACE)-polymerase chain reaction (PCR) based on the transcriptional data of pearl oyster, and then real-time quantitative PCR was used to examine the expression profiles of BP3 gene in different tissues and in the feet with/ without the byssus of pearl oyster. The results showed that the full-length cDNA of BP3 gene was 601 bp, including a 5'untranslated region of 71 bp, a 3'untranslated region of 77 bp, and an open-reading frame of 453 bp, which encodes a deduced protein of 150 amino acids. The molecular weight of BP3 was 16.81 ku, and the isoelectric point was 7.33. BP3 was a hydrophilic stable extracellular protein, mainly located in the extracellular matrix. The sequence comparison showed that BP3 in the pearl oyster shares 30.34% sequence identity with BP3 in mussel
Mytilus coruscus
. The real-time quantitative PCR results showed that BP3 gene was expressed in the tested tissues, with the maximal relative expression level in mantle. There were significant differences among the mantle and feet, adductor muscle, pearl pocket (
P
<0.05), without significant difference among the mantle and gill, lip flap (
P
>0.05). While, the expression level in the feet with byssus was significantly higher than that in the feet without byssus (
P
<0.05) in pearl oyster. These findings could provide new information for further study of the molecular mechanism of byssus adhesion in pearl oyster.
2023 Vol. 42 (4): 613-621 [
Abstract
] (
89
)
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(1 KB)
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(3619 KB) (
56
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622
Dynamic Changes in Physical-Chemical Factors and Microbial Diversity in Ecological Culture Ponds of Largemouth Bass
Micropterus salmoides
ZHANG Guangchen, WANG Yang, SUN Xueliang, LUO Yunlong, WANG Jingru, LIU Fanxiang, JI Yanbin
DOI: 10.16378/j.cnki.1003-1111.21081
To explore the changes of microorganisms and water quality physicochemical indices in ecoculture ponds of largemouth bass
Micropterus salmoides
in July—September and their relationship, the microbial community structure was determine by high throughput sequencing technology and the physico-chemical indices were monitored in surface water in the cultured areas and microecosystem of a largemouth bass ecological culture pond in Tianjin from July to September. After the aquaculture tail water was treated by aquatic vegetables, the surface water entered the micro-ecological filtration area with brush as the matrix, and then the bottom water entered the aquatic vegetable treatment area and finally entered the 300 m
2
culture pond with4000 individuals of largemouth bass for recycling. The results showed that there was always higher α diversity of microbial diversity in the sediment than that in water and brush, with the dominant microorganisms in Proteobacteria in all samples. Cyanobacteria in the water was found to be gradually increased to the dominant species in September, with change in the dominant bacteria in the brush samples with the time of cultivation and with the maximal proportion of Firmicutes in September. The maximal content of ammonia nitrogen was observed in the water in the aquaculture area and the microecological area in August, with the maximal contents of total nitrogen, orthophosphates, permanganate index, and suspended solids (SS), and pH and total alkalinity in September in the two regions. The correlation analysis showed that there was a significant negative correlation between the nitrate nitrogen content in the water and the Rhizobiales Incertae Sedis abundance in the brush. There was significant negative correlation between the content of nitrite nitrogen and the abundance of Rhodobacteraceae in sediment and Norank_o_ _norank_c_ _Nitrospira in brush (
P
<0.05). The findings provide a reference for the regulation of water quality and microorganisms in ecological ponds and the subsequent related studies, and the screening of functional bacteria for remediation of water quality in largemouth bass from July to September
2023 Vol. 42 (4): 622-631 [
Abstract
] (
123
)
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(1 KB)
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(8300 KB) (
68
)
632
Establishment of a Cas13a-RPA Detection Method for the Causative Pathogen
Vibrio splendidus
of Sea Cucumber
Apostichopus japonicus
Skin Ulcerative Syndrome
LIU Ji, CHEN Yanru, HOU Zhumei, XU Dongxue, GU Yuanxue, SONG Wenqi, SONG Yize, XIA Bin
DOI: 10.16378/j.cnki.1003-1111.22036
In order to prevent and control the skin ulcerative syndrome (SUS) of sea cucumber
Apostichopus
japonicus
caused by
Vibrio
splendidus
and maintain the healthy and sustainable development of the sea cucumber aquaculture industry, this study used the characteristic of LwCas13a protein when specific crRNA in LwCas13a binds to the target RNA, its collateral cleavage activity can be activated to degrade non-targeted exogenous RNA. Combined this characteristic with the recombinase polymerase amplification (RPA) technique, a rapid, sensitive and specific quantitative detection method for gyrB gene of
V
.
splendidus
—RPA-Cas13a was established. This method can realize rapid real-time detection of
V
.
splendidus
at constant 37 ℃ within 60 mins. Sensitivity test showed that the detection limit of this method is 550 copies/μL(gyrB), which is consistent with the sensitivity level of fluorescence quantitative PCR (qPCR); Specificity test demonstrated a relatively high specificity with the fact that it had no cross-reactions with other
Vibrio
species, e.g.
V
.
vulnificus
,
V
.
harveyi
and
V
.
anguillarum
, and other common aquatic animal bacterial pathogens, including
Pseudoalteromonas nigrifacien
s,
Aeromonas
hydrophlia
and
Edwardsiella
tarda
. The detection rate of 57 environmental samples including skin mucus of
A
.
japonicus
(32 samples), culture pond seawater (15 samples) and surface sediments (10 samples) was 8.77%, highly consistent with that of traditional fluorescence qPCR method (
Kappa
=1.00) with no statistical difference (
P
>0.05). In conclusion, this study established a novel rapid, simple, sensitive and specific RPA-Cas13a quantitative detection method for
V
.
splendidus
, which provides a powerful molecular tool for preventing and controlling the occurrence of sea cucumber SUS
2023 Vol. 42 (4): 632-639 [
Abstract
] (
94
)
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(1 KB)
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(3660 KB) (
155
)
640
Effects of Acid-Base Stress on Survival Rate,Histopathology and Detoxification-Related Enzyme Activities of Mandarin Fish
Siniperca chuatsi
ZHANG Chengshuo, ZHAO Yan, WANG Yanling, ZENG Mengdong, ZHAO Jinliang
DOI: 10.16378/j.cnki.1003-1111.21146
To investigate the effects of acid-base stress on survival rate,histopathology and detoxification related enzyme (cytochrome P450, flavin-containing monooxygenase, UDP-glucuronosyltransferase, glutathione S-transferase) activities, mandarin fish
Siniperca chuatsi
[body weight: large size (30.2±5.1) g and small size (5.3±0.7) g] were subjected to a 72 h static acute exposure in different pH groups (pH 4.0, 5.5, 7.0, 8.5, 10.0). The results showed that large size fish survived for 72 h in each experimental group. All small size fish died at pH 4.0 and 10.0, and the average survival time was (53.60±4.65) min and (49.30±5.34) min, respectively. After 72 h, except that the morphology of gill and liver tissue in pH 7.0 group was normal, the other experimental groups showed several abnormalities. In the pH 4.0 group, gill capillaries were broken and blood cells accumulated. In the pH 5.5 and pH 8.5 groups, hepatocytes were swollen. In the pH 10.0 group, gill epithelial cells fell off and cytoplasmic vacuolation was more obvious. The activities of cytochrome P450 and flavin-containing monooxygenase increased first and then decreased with the stress time in gill and liver tissue. The activity of glutathione S-transferase was significantly increased and higher than that in the pH 7.0 group after 72 h (
P
<0.05). The activity of UDP-glucuronosyltransferase was lower than that in the pH 7.0 group after 72 h in gill. Taken together, these results suggest that pH 4.0 and pH 10.0 are extremely toxic to mandarin fish fry, acid-base stress cause damage to mandarin fish, cytochrome P450, flavin-containing monooxygenase, UDP-glucuronosyltransferase and glutathione S-transferase participate in the detoxification process of mandarin fish upon acid-base stress, and work differently in time and efficiency.
2023 Vol. 42 (4): 640-647 [
Abstract
] (
143
)
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(1 KB)
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(12361 KB) (
58
)
648
Cloning and Expression Analysis of Myocardial Globulin Binding Protein C3 (MyBPC3) of Jinbian Carp
Cypinus carpio
WEI Lingjing, LYU Yejian, GAN Baojiang, HUANG Jie, LIU Kang, ZHANG Sheng, TENG Zhongzuo, MO Feilong, YE Xiangchen
DOI: 10.16378/j.cnki.1003-1111.21057
In this study, the structure and organizational expression characteristics of myosin binding protein C3 (MyBPC3) gene were investigated in Jinbian carp
Cypinus carpio
with body weight of (74.74±0.92) g. Functions of MyBPC3 on regulation mechanism of skin color variation were analyzed. The full-length cDNA sequence of MyBPC3 gene was cloned by RACE technique, and its tissue expression and correlation with melanin synthesis related genes (TYR, TYRP1, TYRP2 and MC-1R) were analyzed. The results showed that the full length cDNA of MyBPC3 gene was 4253 bp, including an open reading frame (ORF, 3783 bp) encoding 1260 amino acids. The predicted protein had molecular weight of 141.57 ku, the theoretical isoelectric point of 6.42, and 22 PKC and 5 PKA potential phosphorylation sites. Phylogenetic tree and homology analysis showed that the MyBPC3 gene of Jinbian carp was the most closely related to that of crucian carp
Carassius auratus
. The qRT-PCR results indicate that MyBPC3 mRNA is distributed in different tissues of Jinbian carp, with significantly higher expression in heart than that in brain, skin, muscle, spleen, liver, gut, and kidney tissues, and significantly lower expression level in heart and skin tissues of the carp with yellow skin individuals than that in the black one (
P
<0.05). The correlation analysis revealed that the MyBPC3 gene was significant positively correlated with melanin synthesis related genes (TYR, TYRP1, TYRP2 and MC-1R). In conclusion, MyBPC3 gene might involve in melanin synthesis of the skin, and its expression difference might be the cause of the skin color variation of Jinbian carp, which provided a reference for the study of the differential regulation mechanism of skin pigmentation in Jinbian carp.
2023 Vol. 42 (4): 648-656 [
Abstract
] (
87
)
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(1 KB)
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(6193 KB) (
66
)
657
Cloning and Expression Analyses of β-Thymosin in Sea Cucumber
Apostichopus japonicus
SUN Hongjuan, JIANG Jingwei, DONG Ying, GAO Shan, GUAN Xiaoyan, CHEN Zhong, WANG Bai, PAN Yongjia, XIAO Yao, JIANG Pingzhe, LI Peipei, ZHANG Chenyu, ZHOU Zunchun
DOI: 10.16378/j.cnki.1003-1111.21204
β-Thymosin is a polypeptide with antimicrobial activity. In this study, a β-Thymosin was cloned by RACE method from sea cucumber
Apostichopus japonicus
, named Ajβ-Thymosin. The full length of Ajβ-Thymosin was 1877 bp, with an ORF of 126 bp encoding 41 amino acids. The molecular weight of Ajβ-Thymosin was 4.6 ku and theoretical isoelectric point was 5.25. Ajβ-Thymosin contained conserved functional sequences: EVASFDKSKLK and G-actin binding residues: LKKTET. Phylogenetic analysis revealed that Ajβ-Thymosin gathered with sea urchin
Strongylocentrotus purpuratus
β-Thymosin. Quantitative real-Time PCR was selected to examine the expression of Ajβ-Thymosin at different tissues and developmental stages. The result indicated that Ajβ-Thymosin had most highly expression level in coelomocytes and began up-regulated at early auricularia. After four pathogens stimulus, the expression of Ajβ-Thymosin was all inhibited at 4 h. In groups of
Bacillus cereus
and
Shewanella baltica
, the expression of Ajβ-Thymosin was up-regulated at 12 h. The expression of Ajβ-Thymosin was up-regulated at 24 h in the group of
Pseudoalteromonas nigrifacien
and at 48 h in the group of
Vibrio splendidus
, respectively. The dynamic expression patterns of Ajβ-Thymosin at different times in response to different pathogens proved that it participated in the immune reaction in sea cucumber coelomocytes with different immune response abilities. The sequence and expression analyses of Ajβ-Thymosin lay foundations for studying new type antimicrobial peptides and prevention strategy of aquaculture diseases.
2023 Vol. 42 (4): 657-663 [
Abstract
] (
109
)
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(1 KB)
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(3606 KB) (
112
)
664
Population Genetic Study of
Triplophysa yarkandensis
in Tarim River Basin in Xinjiang
ZHAO Wenhao, YI Shaokui, ZHOU Qiong, SHEN Jianzhong, LI Dapeng, ZHOU Xiaoyun
DOI: 10.16378/j.cnki.1003-1111.21211
To understand the population genetic characteristics of
Triplophysa yarkandensis
in Tarim River Basin in Xinjiang, eight populations of
T. yarkandensis
were collected from two tributaries of the Tarim River Basin (Qarqan River and Weigan River), and the genetic diversity and population structure were analyzed using mitochondrial COⅠ gene, as well as the newly developed SSR markers. Based on COⅠ sequences, 22 haplotypes were detected, with average haplotype diversity index of 0.8829 and nucleotide diversity index of 0.0032. Based on SSR markers, there was polymorphism information content of 0.629—0.696, observed heterozygosity of 0.607—0.736 and expected heterozygosity range of 0.655—0.755. The results above suggested a high level of genetic diversity in each population. Both COⅠ and SSR results exhibited lower genetic differentiation between populations; the AMOVA results showed that the genetic variations were primarily derived from intra-population (COⅠ, 95.12%; SSR, 97.23%); the genetic distance exhibited a lower level; and the gene flow in the studied populations were higher than 1, suggesting a weak degree of genetic differentiation and an obviously gene flow between populations. The UPGMA phylogenetic tree constructed based on SSR markers showed that the populations from Qarqan River and Weigan River were clustered into separate genetic branch; there was obviously different genetic composition between populations from the two tributaries based on the population structure analysis; and potential genetic barriers were observed based on Barrier analysis. These results indicated that some genetic variation occurred between populations from the two tributaries. To sum up, the genetic diversity of
T. yarkandensis
populations was found to be relative higher in Tarim River, and obvious gene flow between populations can be detected. However, because of the habitat fragmentation in Tarim River system, potential gene flow barriers occurred between populations from Qarqan River and Weigan River, which resulted in obvious genetic differentiation between populations from the two tributaries. Thus, it is necessary to take effective measure to protect the habitat environment of
T. yarkandensis
to avoid the potential loss of genetic diversity.
2023 Vol. 42 (4): 664-673 [
Abstract
] (
98
)
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674
Structural and Functional Characteristics of Intestinal Microbiota in Different Populations of
Macrobrachium rosenbergii
Larvae
SHI Jingu, WU Xia, HUANG Guanghua, YANG Huizan, LIANG Yi, LYU Min, ZENG Lan, HU Dasheng, HUANG Libin, WANG Rui
DOI: 10.16378/j.cnki.1003-1111.21068
Under the same culture conditions, five populations of
Macrobrachium rosenbergii
were cultured. The larvae of
M. rosenbergii
hatched for 20 days in the same batch were selected. The intestinal microflora of the larvae of Burmese population, Sri Lankan population, Vietnamese population, Bengali population and own variety 1 population were analyzed by 16S rRNA high-throughput sequencing technology. The results showed that there were 256 OTUs in common in five groups, and the unique OTUs of each group were 131, 247, 231, 144 and 184, respectively. The species richness of intestinal microbiota in Bengali population was the highest, and the species diversity of gut microbiota in Vietnamese population was the highest. The dominant microflora of five groups were similar in phylum and family level, which were Proteobacteria, Bacteroidetes and Firmicutes, but their abundance and composition of other microflora were different. The abundance of Firmicutes in intestinal microflora of Sri Lankan population was higher than that of Bacteroidetes, which was different from the other four groups. The five groups also have common functions, mainly focusing on carbohydrate metabolism, amino acid metabolism, vitamin metabolism and so on. The results showed that the intestinal microbiota of different populations of
M. rosenbergii
had some similarities in structure, abundance and metabolic function. This study analyzed the structure and functional characteristics of intestinal microbiota of different populations of
M. rosenbergii
larvae, which laid a foundation for further study on the growth and development mechanism of different populations of
M. rosenbergii
.
2023 Vol. 42 (4): 674-681 [
Abstract
] (
105
)
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682
Observation of the Development of Embryos, Larvae and Juveniles of
Tapes dorsatus
ZHANG Kexin, YANG Shangsong, LUO Zexin, ZHANG Yuan, ZHAN Jianqiang, CHEN Linguang, LU Yining, LIU Zhigang
DOI: 10.16378/j.cnki.1003-1111.21125
In order to study the morphological characteristics and development rate of
Tapes dorsatus
in its early development stage, and to provide reference for the morphological development biology and artificial seeding of
T. dorsatus
, we observed, measured and photographed each developmental stages of the seeding process using a light microscope under water temperature of 25—26 ℃ and salinity of 29—31. The results showed that the newly fertilized eggs were round without blastocysts, and the size was (59.0±2.2) μm. After fertilization, the thin and bright fertilization membrane was raised and the polar body was released. The shell length of D-shaped larvae was (101.8±3.7) μm. The hinge line of the early umbo larvae was slightly curved and spherical foot primordium appeared. The umbo of medium term umbo larvae continue to uplift and foot primordium developed into boot foot shape, the base of the foot primordium formed a pair of statocysts which were 10 μm in diameter; in the post stage of umbo larvae, the umbo of larvae was prominent, and the boot-like foot extended into a stick, forming two pairs of tubular gill filaments on both sides of the foot base. The mature larvae had no “eye spots”, statocysts grew to 15 μm. Metamorphosis larvae's shell length was (228.4±11.2) μm, and velum shrinked and disappeared. Simultaneously secreted one or two byssus on the bottom of the foot, and the life of the larvae changed from planktonic life to attached life. In this stage, metamorphosis larvae secreted secondary shell to become juveniles. In the non siphon stage, due to the asymmetric growth of the secondary shell, the top of the shell was protrusive and inclined forward, and the gill filaments increased. In the single siphon stage, an outlet pipe appeared above the shell of juvenile. In the double siphon stage, the juvenile grew out of the inlet pipe, and the length was shorter than that of the outlet pipe. The end of the inlet pipe was surrounded by tentacles. The shell shape was basically the same as that of the adult shellfish, and the front end was rounded and the back end was trunk-shaped.The shell length of double siphon juvenile was (578.1±76.1) μm. It was found that the growth rule of the clam was “fast-slow-fast”. Larvae took 16 days to develop from the D-shaped stage to the metamorphosis, and juvenile took 30 days to develop from the attached life stage to the double siphon stage.
2023 Vol. 42 (4): 682-689 [
Abstract
] (
86
)
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182
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690
Catching Performance of Combination of Square and Diamond Mesh Codend of Trawl in the Northern Part of South China Sea
YAN Lei, YANG Bingzhong, LI Yanan
DOI: 10.16378/j.cnki.1003-1111.21168
In order to evaluate the mesh selectivity of combination of square and diamond mesh codend for the main catches of trawl in the northern South China Sea, the captive selectivity was comparatively investigated in combined mesh codend (treatment net group) and traditional-diamond mesh codend (control net group) using the covered net method, and the selectivity curves were fitted with Logistic and Richards curves to analyze the selectivity performance of 2 different mesh structure codends for blue round scad
Decapterus maruadsi
in the northern South China Sea. The results showed that there were eight species with number escape rate and biomass escape rate higher than 80% in the treatment net group, while there were only five species in the control net group. As the highest quality catch species in the two mesh codends, the number escape rate and mass escape rate of blue round scad were found to be 25.35% and 20.98% in the treatment net group, and 16.39% and 15.48% in the control net group, respectively. The selectivity showed that the Richards curves of the two nets were better fitted, with smaller model residuals and Akaike's information index (AIC values), and the 50% retention lengths (
L
50
) for blue round scad were 14.31 cm and 13.60 cm, respectively. In summary, there were higher escape rates in the reatment net for a variety of bycatch. The findings indicated that application of the combined square and diamond mesh codend effectively improved the trawl selectivity and protected the sustainable use of fishery resources.
2023 Vol. 42 (4): 690-697 [
Abstract
] (
112
)
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62
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698
Heritability Estimating and Correlativity of Growth Traits in Juvenile Edible Jellyfish
Rhopilema esculentum
LI Yulong, ZHANG Weijie, TIAN Meilin, SUN Wei, CHEN Bailing, ZHANG Chenyu, ZHOU Zunchun, LI Yunfeng
DOI: 10.16378/j.cnki.1003-1111.21154
China is the only one country for farming of edible jellyfish
Rhopilema esculentum
. With the expansion and intensification of the jellyfish, various questions such as diseases, poor quality of larvae and less natural resources, have occurred and been serious, causing economic losses. At last, these questions are resolved by provided excellent breeds provided by using the selecting breeding program. It is an important way of selecting breeding. In this study, the phenotypic values of growth traits including body weight and umbrella diameter were measured in 700 individuals (300 days old) of edible jellyfish from 14 full-sib families to estimate the heritability and correlativity of growth traits. The results showed that the heritability of 300 days old individuals was 0.843±0.198 in body weight, and 0.426±0.098 in umbrella diameter, the heritability of these two traits being within higher heritability. In the 300 days old jellyfish, the genetic correlation and phenotypic correlation between body weight and umbrella diameter were 0.962±0.022 and 0.911±0.018 respectively, and both with highly positive genetic correlations. The regression analysis of phenotypic values of growth traits including body weight and umbrella diameter from 700 samples of 300 days old jellyfish revealed the mathematical model of growth traits [body weight (
y
) and umbrella diameter (
x
)]:
y
= 0.1624
x
2.7291
(
r
2
=0.8815). The findings provide guidance to the indirect selection and pre-selection in selective breeding of the jellyfish
R. esculentum
.
2023 Vol. 42 (4): 698-704 [
Abstract
] (
110
)
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135
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705
Substance Exchange between Water-Vascular System and Coelom in Sea Cucumber
Apostichopus japonicus
FAN Xuyuan, WANG Zhenhui, REN Yuan, GUO Liyuan, SUN Qirui, WANG Yinan, LI Qiang
DOI: 10.16378/j.cnki.1003-1111.21199
Sea cucumber
Apostichopus japonicus
with a body mass of (65±10) g was injected in coelom with beet root pigments—beetroot red, purified and adjusted GST-tagged protein, fluorescent microsphere dilution, suspension of
Vibrio splendidus
and suspension of
Escherichia coli.
The distribution of the injected matters in the coelom and the water-vascular system (represented by polian vesicle) was detected to explore the correlation, connectivity and material exchange between the coelom and the water-vascular system of the sea cucumber. The results showed that the coelomic fluid from coelom and polian vesicle (hereinafter referred to as “vesicle fluid”) were stained into red at 6 hours after injection by beet red dye. A large number of GST-tag proteins were detected in the coelomic fluid, and there were also some proteins in the vesicle fluid 6 h and 12 h after protein injection. Many fluorescent microspheres were observed in the coelomic fluid, but not in the vesicle fluid 6 h and 12 h after injection. In addition, a large number of
E. coli
and
V. splendidus
bacteria were found in the coelomic fluid 6, 12, 24 and 72 h after injection, without bacteria isolated in the vesicle fluid. In conclusion, the water-vascular system and the coelom are not found to be infinitely unblocked in sea cucumber. Soluble substances such as small molecules and biological macromolecules were exchanged between the coelom and the water-vascular system, but the exogenous particles and bacteria did not enter the water-vascular system via the coelom. The findings laid a theoretical foundation for the subsequent study of the physiological structure and biological role of the water-vascular system in sea cucumber.
2023 Vol. 42 (4): 705-711 [
Abstract
] (
108
)
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47
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712
Effects of Dietary
Bacillus velezensis
MSP05 on Growth Performance, Intestinal Digestive Capacity and Non-Specific Immune of Loach
Misgurnus anguillicaudatus
MENG Wenrong, MI Haoyu, HUANG Yichen, HUANG Zhifa, LIU Qitong, XIA Mengjie, FANG Yaowei
DOI: 10.16378/j.cnki.1003-1111.21127
This study was conducted to investigate the effect of dietary
Bacillus velezensis
MSP05 on growth performance, digestion, and non-specific immune function of juvenile loach
Misgurnus anguillicaudatus
. 360 fishes with an average initial body weight of (3.24±0.14) g were randomly assigned into 4 groups in triplicates with 30 individuals per replicate. One control group and three experimental groups were added 0, 1×10
6
, 1×10
7
, 1×10
8
cfu/g
B. velezensis
MSP05 to basal diets respectively. After 8 weeks of feeding, the results demonstrated that the weight gain rate(WGR) and specific growth rate(SGR) of the groups adding
B. subtilis
MSP05 were significantly higher than those of the control group (
P
<0.05). The activities of protease and amylase increased firstly and then decreased with the
B. velezensis
level increasing, achieving the highest value in 1×10
7
cfu/g group, while the activities of lipase rose continuously. The optimal morphology of foregut, midgut and hindgut of loach were observed in 1×10
6
, 1×10
6
, 1×10
7
cfu/g group, respectively. The activities of serum catalase (CAT), superoxide dismutase (SOD), lysozyme (LSZ) and alkaline phosphatase (ALP) in the serum were significantly higher than the control group (
P
<0.05), while MDA levels showed the opposite trend. Challenged by
Aeromonas hydrophila
for 15 days, the cumulative mortality rate of juvenile loaches decreased significantly fed with
B. velezensis
. In conclusion, the addition of
B. velezensis
MSP05 can significantly enhance the growth performance, digestive enzymes activity, intestinal morphology, antioxidant enzyme activity, non-specific immune function and disease resistance against
A. hydrophila
.
2023 Vol. 42 (4): 712-718 [
Abstract
] (
96
)
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123
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719
Inhibitory Effect of Dimethylsulfoxide on
N
-Acetyl-β-
D
-Glucosaminidase from Chinese Horseshoe Crab
Tachypleus tridentatus
LIN Jiancheng, WU Jianhong, LIN Juanjuan
DOI: 10.16378/j.cnki.1003-1111.21130
The periodic molting of arthropods is closely related to
N
-acetyl-β-
D
-glucosaminidase (EC3.2.1.52, NAGase). NAGase was extracted from the viscera of
Tachypleus tridentatus
. The effect of dimethyl sulfoxide (DMSO) on NAGase was studied by using the kinetics method of enzymatic reaction. The results showed that DMSO had strong inhibitory effect on the NAGase, the enzyme activity decreased by 38.0% when its concentration reached 0.1 mol/L. The inhibitory effect of DMSO on NAGase was reversible. The kinetics showed that the inhibitory effect of DMSO solution on NAGase was the competitive and non-competitive mixed type. The inhibition constants of DMSO on free enzyme (
K
I
) and on enzyme-substrate complex(
K
IS
) were determined to be 0.200 mol/L and 1.063 mol/L, respectively. The value of
K
I
was less than
that of
K
IS
, indicating a marked protective effect of the substrate on the inactivation of the enzyme. As the concentration of substrate increased, the inactivation of enzyme in DMSO turned to decrease. Meanwhile, UV absorption spectra and fluorescence emission spectra of NAGase were measured. The UV absorption of NAGase treated by DMSO was enhanced with increasing DMSO concentration, but the fluorescence emission peak intensity of the enzyme gradually decreased, their UV absorption peak and fluorescence emission peak of NAGase did not shift. The fluorescence emission of enzyme was almost quenched when the concentration of DMSO reached 0.65 mol/L. It showed that changes of enzyme conformation in the DMSO solvent led to inactivation of the NAGase. The above results confirmed that DMSO might further affect its molting by inhibiting the activity of NAGase from
Tachypleus tridentatus
.
2023 Vol. 42 (4): 719-725 [
Abstract
] (
82
)
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71
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Overview and Specific Topi
726
Advances on Epigenetic Mechanism of Sex Differentiation in Teleost Fish
JIANG Jieming, LIU Ying, LIU Qi, YAN Hongwei
DOI: 10.16378/j.cnki.1003-1111.22010
2023 Vol. 42 (4): 726-734 [
Abstract
] (
150
)
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(1111 KB) (
132
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735
Research Progress and Development Trend in RecirculatingAquaculture System
WANG Haiheng, HOU Haochen, LIU Ying
DOI: 10.16378/j.cnki.1003-1111.21096
2023 Vol. 42 (4): 735-741 [
Abstract
] (
298
)
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(1078 KB) (
177
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742
Research Progress of Germplasm Innovation on
Eriocheir sinensis
XING Xiumei, SHEN Minghao, LI jie, NING Xianhui, ZHANG Kai, YIN Shaowu
DOI: 10.16378/j.cnki.1003-1111.21097
2023 Vol. 42 (4): 742-746 [
Abstract
] (
158
)
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(1 KB)
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(1012 KB) (
248
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