Application of otolith shape analysis for stock discrimination and species identification of five goby species （Perciformes： Gobiidae） in the northern Chinese coastal waters

We tested the use of otolith shape analysis to discriminate between species and stocks of five goby species( Ctenotrypauchen chinensis, Odontamblyopus lacepedii, Amblychaeturichthys hexanema, Chaeturichthys stigmatias, and Acanthogobius hasta) found in northern Chinese coastal waters. The five species were well differentiated with high overall classification success using shape indices(83.7%), elliptic Fourier coefficients(98.6%), or the combination of both methods(94.9%). However, shape analysis alone was only moderately successful at discriminating among the four stocks(Liaodong Bay, LD; Bohai Bay, BH; Huanghe(Yellow) River estuary HRE, and Jiaozhou Bay, JZ stocks) of A. hasta(50%–54%) and C. stigmatias(65.7%–75.8%). For these two species, shape analysis was moderately successful at discriminating the HRE or JZ stocks from other stocks, but failed to effectively identify the LD and BH stocks. A large number of otoliths were misclassified between the HRE and JZ stocks, which are geographically well separated. The classification success for stock discrimination was higher using elliptic Fourier coefficients alone(70.2%) or in combination with shape indices(75.8%) than using only shape indices(65.7%) in C. stigmatias whereas there was little difference among the three methods for A. hasta. Our results supported the common belief that otolith shape analysis is generally more effective for interspecific identification than intraspecific discrimination. Moreover, compared with shape indices analysis, Fourier analysis improves classification success during inter- and intra-species discrimination by otolith shape analysis, although this did not necessarily always occur in all fish species.     

Comparative study of otolith and sulcus morphology for stock discrimination of yellow drum along the Chinese coast

Otolith morphology is widely used for fish stock identification.The sulcus,a structure on the medial side of the otolith,is an important feature in morphological analysis.This study was conducted to evaluate the feasibility of using sulcus morphology for stock identification and to compare its performance with commonly used otolith morphology analysis.Otoliths were collected and analyzed from three geographical groups(the Huanghe(Yellow)River estuary,HHE;the Jiaozhou Bay,JZB;and the Changjiang(Yangtze)River estuary,CJE)of yellow drum Nibea albiftora.The results show that the analysis of sulcus morphology based on shape indices(SIs),elliptic Fourier coefficients(EFc),and a combination of the two parameters identified stocks at overall classification rates of 51.0%,72.5%,and 73.2%,respectively.These classification rates are similar to those obtained using otolith morphology analysis(57.0%,73.8%,and76.5%by SIs,EFc,and their combination,respectively).The findings suggest that sulcus morphology is comparable to the commonly used otolith morphology for identifying stocks of sciaenids,such as the yellow drum.For both otolith and sulcus morphology,EFc could identify the stocks more efficiently than SIs,while the combination of SIs and EFc was even better.     

Otolith shape analysis for stock discrimination of two Collichthys genus croaker(Pieces: Sciaenidae,) from the northern Chinese coast

The otolith morphology of two croaker species(C ollichthys lucidus and C ollichthys niveatus) from three areas(Liaodong Bay, LD; Huanghe(Yellow) River estuary, HRE; Jiaozhou Bay, JZ) along the northern Chinese coast were investigated for species identifi cation and stock discrimination. The otolith contour shape described by elliptic Fourier coefficients(EFC) were analysed using principal components analysis(PCA) and stepwise canonical discriminant analysis(CDA) to identify species and stocks. The two species were well dif ferentiated, with an overall classifi cation success rate of 97.8%. And variations in the otolith shapes were significant enough to discriminate among the three geographical samples of C. lucidus(67.7%) or C. niveatus(65.2%). Relatively high mis-assignment occurred between the geographically adjacent LD and HRE samples, which implied that individual mixing may exist between the two samples. This study yielded information complementary to that derived from genetic studies and provided information for assessing the stock structure of C. lucidus and C. niveatus in the Bohai Sea and the Yellow Sea.     

Stock discrimination of spottedtail goby (Synechogobius ommaturus) in the Yellow Sea by analysis of otolith shape

Otolith shape is species specific and is an ideal marker of fish population affiliation. In this study, otolith shape of spottedtail goby Synechogobius ommaturus is used to identify stocks in different spawning locations in the Yellow Sea. The main objectives of this study are to explore the potential existence of local stocks of spottedtail goby in the Yellow Sea by analysis of otolith shape, and to investigate ambient impacts on otolith shape. Spottedtail goby was sampled in five locations in the Yellow Sea in 2007 and 2008. Otoliths are described using variables correlated to size (otolith area, perimeter, length, width, and weight) and shape (rectangularity, circularity, and 20 Fourier harmonics). Only standardized otolith variables are used so that the effect of otolith size on the shape variables could be eliminated. There is no significant difference among variables of sex, year, and side (left and right). However, the otolith shapes of the spring stocks and the autumn stocks differ significantly. Otolith shape differences are greater among locations than between years. Correct classification rate of spottedtail goby with the otolith shape at different sampling locations range from 29.7%–77.4%.     

Using otolith weight to predict the age of Pennahia macrocephalus in the mouth of the Beibu Gulf

The relation between otolith weight (OW) and the age of marine fish is studied. A total of 222 individuals of bighead white croaker, Pennahia macrocephalus were sampled seasonally in the mouth of the Beibu Gulf, the South China Sea, in 2007. Since there are no significant differences in sagittal OW between otolith in pairs (P≧>0.05), the undamaged left sagittal otolith is used for age determination. The highest correlations among standard length, OW and fish ages are confirmned by linear, exponential and multinomial regression. Results show that sagittal OW overlaps only occasionally among age groups, and to individuals with similar standard length, the older and slower-growing fish has a heavier otolith because of the continued otolith material deposition. There are differences in sagittal OW among different age groups and significant positive linear relationship with age (P<0.05). The age readings can be verified by plotting the sagittal OW versus the standard length for age groups, and the individuals with similar standard length but in different ages can be separated by sagittal OW frequency analysis. Mostly, the predicted ages using the regression between sagittai OW and ages are closed to the observed ages by counting annulus on scale. It indicates that the sagittal OW analysis is a useful technique for validating the accuracy of age determination by annuli counts, especially for individuals of similar size. Furthermore,the technique is applied for Pennahia macrocephalus with discussion in this paper.     

Use of Otolith Shape for the Identification of Trumpeter Sillago （Sillago maculata） and Silver Sillago （Sillago sihama）

Otolith shape is species-specific in fish. In this study, 161 samples of Trumpeter sillago Sillago maculata and 164 of Silver sillago Sillago sihama were collected from Beibu Gulf in July 2009. The main objective of this study was to use morphological variables and Fourier harmonics to describe otolith characteristics and use discriminant analyses to separate the two species. Otoliths were measured by traditional one and two dimensional measures(otolith length, width, diameters, area, perimeter, rectangularity and circularity), as well as by Fourier analysis to capture the finer regions of the otolith. Analysis of covariance(ANCOVA)showed that there was significant correlation between morphological variables(diameter, perimeter, otolith length, otolith width, otolith area, density, harmonics 3, harmonics 4, harmonics 5, harmonics 13, harmonics 15, and harmonics 16)and body length. To minimize size effects on the morphological variables between species, only fish with a body length between 90 and 140 mm were included in the data analysis and the variables which had significant relation with body length were transformed using the residual. The result showed that the accuracy of discriminant analysis was 97. 8% and 100% for Trumpeter sillago and Silver sillago, respectively. This high accuracy indicated that otolith shape was described accurately by morphological variables and Fourier harmonics, and discriminant analysis was an effective way to identify and separate the two species.     

Using otolith weight to predict the age of <Emphasis Type="Italic">Pennahia macrocephalus</Emphasis> in the mouth of the Beibu Gulf

The relation between otolith weight (OW) and the age of marine fish is studied. A total of 222 individuals of bighead white croaker, Pennahia macrocephalus were sampled seasonally in the mouth of the Beibu Gulf, the South China Sea, in 2007. Since there are no significant differences in sagittal OW between otolith in pairs (P≥0.05), the undamaged left sagittal otolith is used for age determination. The highest correlations among standard length, OW and fish ages are confirmed by linear, exponential and multinomial regression. Results show that sagittal OW overlaps only occasionally among age groups, and to individuals with similar standard length, the older and slower-growing fish has a heavier otolith because of the continued otolith material deposition. There are differences in sagittal OW among different age groups and significant positive linear relationship with age (P<0.05). The age readings can be verified by plotting the sagittal OW versus the standard length for age groups, and the individuals with similar standard length but in different ages can be separated by sagittal OW frequency analysis. Mostly, the predicted ages using the regression between sagittal OW and ages are closed to the observed ages by counting annulus on scale. It indicates that the sagittal OW analysis is a useful technique for validating the accuracy of age determination by annuli counts, especially for individuals of similar size. Furthermore, the technique is applied for Pennahia macrocephalus with discussion in this paper. Supported by the National Natural Science Foundation of China (No. 30771653) and Bureau of Fisheries, Ministry of Agriculture     

Analyses of egg size,otolith shape,and growth revealed two components of small yellow croaker in Haizhou Bay spawning stock

The geographic al variations in life history characteristic s of small yellow cro aker Larimich thys polyactis,caused by experienced different environmental conditions,have been observed in China seas.Previous studies based on spatial distribution,migration route,and body morphometrics suggested a complex stock structure.In this study,to clarify the source of a spawning stock,we investigated the reproduction strategy and inter-structure of the Haizhou Bay(HZB)spawning stock in the middle Yellow Sea from both egg survey and adult otolith increment analysis.Egg and adult samples were collected from three surveys during spawning season in 2013.Distinct spatial and temporal variations were detected in egg distribution and size,and otolith shape analy sis of adult fishes revealed two morphotypes with different increment growth using random forest cluster.The results indicate the existence of two components within the same spawning stock in HZB from different wintering grounds,and accordingly special protection should be required for this stock given the significance to maintain connectivity between adj acent subpopulations.     

Population identification of red-fin pargo,Parargyrops edita Tanaka,in Taiwan strait and Beibu Gulf

This paper deals with population identification ofParargyrops edita Tanaka. Samples of spawning stocks were collected from Niushan-Penghu, Minnan-Taiwan shoal (Southern Fujian-Taiwan shoal) and Beibu Gulf during the spawning period 1981–1982. Morphological characters, both meristic and morphometric, are counted and measured respectively. The stock concept and intraspecific categories have been discussed. Some meristic characters as the number of left or right ventral fin spine, right ventral fin rays and abdominal vertebrae of this fish may be constant, others as the numbers of left or right pectoral fin rays and gill-rakers may have obvious geographical variation. Morphological characters have not attained the level of subspecific distinctness, no sub-species could be found among these stocks by means of coefficient of difference (C.D.). Basing upon the comparative analysis ofM _diff, the authors consider that stocks of Minnan-Taiwan shoal and Niushan-Penghu belong to the same local population, Taiwan Strait local population. By means of the discriminant function analysis and linear relationship analysis, statistical differences will be found to exist in meristic characters between the stock of Minnan-Taiwan shoal and that of Niushan-Penghu, so that the two are virtually different stocks or subpopulations. The stock of Beibu Gulf belongs to another called the Beibu Gulf local population. These two local populations are of allopatric distribution and show statistically significant difference between themselves. Among these stocks, the Minnan-Taiwan shoal stock is strongly influenced by the much more complicated environmental conditions of the mixed boundary region of the subtropical Sino-Japanese Subregion in the north and the tropical Indo-Malayan Subregion in the south. Thus its coefficient of variability (C.V.) is greater than those of the other two in the mean values of various meristic characters. This paper was published in Chinese inOceanologia et Limnologia Sinica 14 (6): 511–520.     

Otolith development in larval and juvenile Schizothorax davidi: ontogeny and growth increment characteristics

Laboratory-reared Schizothorax davidi larvae and juveniles were examined to assess the formation and characteristics of David’s schizothoracin otoliths.Otolith development was observed and their formation period was verifi ed by monitoring larvae and juveniles of knownage.The results revealed that lapilli and sagittae developed before hatching,and the first otolith increment was identified at 2 days post hatching in both.The shape of lapilli was relatively stable during development compared with that of sagittae;however,growth of four sagittae and lapilli areas was consistent,but the posterior area grew faster than the anterior area and the ventral surface grew faster than the dorsal surface.Similarly,the sum length of the radius of the anterior and posterior areas on sagittae and lapilli were linearly and binomially related to total fish length,respectively.Moreover,daily deposition rates were validated by monitoring knownage larvae and juveniles.The increase in lapilli width was 1.88±0.080 0μm at the ninth increment,which reached a maximum and the decreased gradually toward the otolith edge,whereas that of sagittae increased more slowly.These results illustrate the developmental biology of S.davidi,which will aid in population conservation and fish stock management.     

Otolith Microstructure Analysis Reveals Different Growth Histories of Japanese Sardine (Sardinops melanostictus) in the Oyashio Waters

After decades of low year classes,the stock of Japanese sardine(Sardinops melanostictus)has begun to recover since the mid-2000s.The hatch dates and otolith growth rates of age-0 juvenile sardine,which were collected in the subarctic Oyashio waters in autumn 2018,were determined from an otolith microstructure analysis.The sardines were hatched from late January to late April,while mostly in February and March.The otolith growth rate increased continuously up to 60 d after hatching and thereafter de-creased.The revealed growth rate in a crucial growth period is faster than that reported for juvenile sardines collected in the 1990s,which is coincided with the recent recovery trend of the sardine stock.Two groups with different hatch dates,growth histories,and migration routes were identified using unsupervised random forest clustering analysis.They were considered inshore and offshore migration individuals in accordance with recent researches.In the offshore group,a high proportion of sardine juveniles hatched late and grew faster in the Kuroshio-Oyashio transitional waters,a finding consistent with the hypothesis of growth-rate-dependent re-cruitment.This finding on the population composition and growth rate of juvenile sardine in the Oyashio waters can be a basis for an improved prediction of their survival and provides us with valuable information on the recruitment processes of this stock during the period of stock recovery.     

Genetic variation in two sea cucumber （Apostichopus japonicus） stocks revealed by ISSR markers

Sea cucumber Apostichopus japonicus samples were collected in Changdao, Penglai (PL), 27 individuals, and Lingshandao, Qingdao (QD), 30 individuals, in the Shandong Peninsula, China. Ten SSR primers were used to assess the genetic variation and relationship between and within the two stocks. Respectively, for each stock, the percentage of polymorphic bands was 85.2% and 86.9%; the gene diver- sity was 0.360 5 and 0.342 8; and the Shannon’s information index was 0.515 0 and 0.499 0. At species level, the percentage of polymorphic bands was 92.2%, the total gene diversity was 0.378 9 and the Shannon’s information index was 0.550 4. The coefficient of overall genetic differentiation and the ge- netic distances between the stocks were also calculated to be 0.073 0 and 0.079 6 using the POPGENE program. Results show that the genetic diversity of the two stocks is still large but the genetic distance between the two stocks is close. A dendrogram was constructed for the 57 individuals from the two stocks, showing that the genetic structure was unitary for PL stock but complex for QD stock.     

Otolith chemical signatures of spottedtail goby Synechogobius ommaturus in coastal waters of China

This study explores the potential existence of local stocks of Synechogobius ommaturus using otolith chemical signatures. The concentrations of 10 elements (K, Ca, Na, Mg, Sr, Pb, Ba, Mn, Co, Zn) in the otoliths from eight stocks (Dandong, Dalian, Tianjin, Dongying, Weihai, Qingdao, Ganyu and Xiamen) of S. ommaturus were measured by Inductively Coupled Plasma Mass Spectrometry and regressed against otolith weight. No significant correlation between otolith weight and concentrations of Na, Mg, Ca, Sr, Co, Ba, Zn and Pb were observed. Concentrations of K and Mn were correlated with otolith weight. Consequently, the residuals of the regressions were used instead of the original concentrations in subsequent analyses. Analysis of variance (ANOVA) showed no significant difference between males and females in otolith chemical signatures. Differences were found among the locations for K, Sr, Mn, Co, Ba, Pb, but not for Ca, Na, Mg and Zn. Residuals of the regressions of elemental concentrations against otolith weight that fitted the normal distribution and homogeneity were studied by discriminant analysis. The success of the discriminant function to predict the location of S. ommaturus capture ranged from 100% for Weihai and Xiamen stocks, to only 44.4% and 62.5% for Qingdao and Dandong stocks, respectively. The average prediction success was 80.3%.     

Single nucleotide polymorphisms in intron 1 and intron 2 of Larimichthys crocea growth hormone gene are correlated with growth traits

The growth hormone gene (GH) affects animal growth and is a potential target for genetic studies of variation related to growth traits. In this study, we analyzed single nucleotide polymorphisms (SNPs) in GH intron regions and their associations with growth traits in large yellow croaker, Larimichthys crocea, from Zhejiang and Fujian stocks. The results of PCR-single strand conformation polymorphism showed two haplotypes of intron 1, named AA and AB genotypes, in Zhejiang stock. AB exhibited an SNP at position 196 (G A) that was negatively correlated with body height and positively correlated with standard length/body height (P 0.05). Two different genotypes, CC and CD, were identified in intron 2 in Fujian stock, with CD showing an SNP at position 692 (T C). The CD genotype had a significantly positive correlation with both weight and total length (P 0.01). These basic data highlight the potential for using GH as a genetic marker of fish growth in marker assisted selection.     

Genetic variation of natural and cultured stocks of Paralichthys olivaceus by allozyme and RAPD

Population genetics of the left-eyed flounder, Paralichthys olivaceus, including natural and cultured stocks distributed in the coastal waters near Qingdao of eastern maritime China, was analyzed in allozyme and RAPD. The results showed that among total 29 gene loci of 15 isozymes, 9 and 7 were po- lymorphic in natural and cultured stocks, respectively. The status of genetic diversity in P. olivaceus is low in terms of polymorphic loci in chi-square test and genetic departure index of Hardy-Weinberg equi- librium. More alleles in IDHP, CAT, GDH and Ldh-C allozymes were found in the fish, which could be used as markers in assortive breeding and distinguishing stock, population or species evolution. Total 88 and 86 RAPD bands ranging from 200 to 2 500 bp were recognized individually in average of 7.8–8.0 bands per primer. The genetic diversity in cultured stock is lower than that in natural ones showing an ob- viously decreasing genetic divergence. Therefore, effective countermeasures must be taken to protect ge- netic resources of marine cultured fishes. The 2 markers have their own pros and cons. Combining the 2 markers to investigate the genetic variation of populations is suggested. The results provide basic data of this flounder and they are useful for studying genetic improvement and genetic resources of the fish.     

Fish community diversity in the middle continental shelf of the East China Sea

The status of fishery stocks in the coastal waters of China is far from ideal, mainly due to climate change and the impacts of human activities (e.g., pollution and overfishing). Thus, the restoration and protection of fishery resources have become critical and complex. The stability and balanced structure of the fish community is a basic foundation for the protection of fishery resources. Based on data collected from bottom trawls by the R/V Beidou in continental shelf of the East China Sea in November 2006 and February 2007, changes in the composition and diversity of fish species and functional groups were analyzed. The research area was divided into offshore waters and inshore waters by the two-way indicator species analysis (TWIA). The results showed that the dominant species were different between offshore waters and inshore waters and also varied with the survey time. The most abundant family was Sciaenidae and Teraponidae in November 2006, Sciaenidae, Engraulidae and Triglidae were most abundant in February 2007. The species belonged mainly to mobile piscivores (G6), benthivores/piscivores (G4), benthivores (G3) and planktivores (G1), and the dominant species in November 2006 were commercial species (e.g. Larimichthys polyactis and Trichiurus japonicus), but small-sized species were dominant in February 2007 (e.g., Harpadon nehereus, Benthosema pterotum, Champsodon capensis, and Acropoma japonicum). The species diversity showed a similar trend as the functional group diversity. Stations with higher diversity were mainly distributed in inshore waters in February 2007, whereas higher diversity was found in offshore waters in November 2006. The highest biomass and species number were found in G6 group, followed by the G4, G5 and G1 groups. The distribution of the number of individuals of each functional group showed the opposite trend as that of the biomass distribution. In addition, the size spectra were mainly concentrated around 3–29 cm, and the individual number of fish species gradually decreased with the increase in body size, but the relative biomass showed a moderate fluctuation in each size class. These changes showed that species with faster growth rate and earlier maturity age became dominant in continental shelf of the East China Sea. So the variations in biological characteristics of fish should be fully considered in maintaining sustainable utilization of fishery resource.     

Effects of Trawl Selectivity and Genetic Parameters on Fish Body Length Under Long-Term Trawling

Long-term fishing pressure affects the biological characteristics of exploited fish stocks. The biological characteristics of hairtail(Trichiurus lepturus) in the East China Sea are unable to recover because of long-term trawling. Fishing induces evolutionary effects on the fish's biological characteristics. Evidence of these changes includes small size at age, a shift to earlier age structure, and early maturation. Natural and artificial selection usually affect the fish's life history. Selection can induce different chances of reproduction, and individual fish can give a different genetic contribution to the next generation. In this study, analysis of time-dependent probability of significance and test of sensitivity were used to explore the effects of fish exploitation rate, mesh size, and heritability with long-term trawling. Results showed that fishing parameters were important drivers to exploited fish population. However, genetic traits altered by fishing were slow, and the changes in biological characteristics were weaker than those caused by fishing selection. Exploitation rate and mesh size exhibited similar evolutionary trend tendency under long-term fishing. The time-dependent probability of significance trend showed a gradual growth and tended to be stable. Therefore, the direction of fishing-induced evolution and successful management of fish species require considerable attention to contribute to sustainable fisheries in China.     

华贵栉孔扇贝两种壳色群体生长和消化酶活力比较

以华贵栉孔扇贝桔黄和紫褐壳色个体为亲本建立两个壳色群体,从壳长、壳高、壳宽和体重几方面进行生长比较,并对两个群体做了淀粉酶和纤维素酶活力比较。结果表明:桔黄壳色群体的平均壳长、平均壳高、平均壳宽和平均体重显著大于紫褐壳色群体(P<0.05)。在24℃、27℃、30℃和33℃条件下,桔黄壳色群体的淀粉酶和纤维素酶活力显著高于紫褐壳色群体(P<0.05);不同壳色群体生长与其淀粉酶和纤维素酶活力有关。     

Monthly variation in the fat content of anchovy (Engraulis japonicus) in the Yellow Sea: implications for acoustic abundance estimation

Anchovy is a key species in the Yellow Sea ecosystem. An accurate estimate of anchovy abundance is vital for the management of the anchovy stock and measurement of the ecosystem response to changes in anchovy abundance. However, the acoustic fish abundance estimate may be biased by 30%?C40% if the fat-content induced target strength variation is not taken into account. We measured the monthly variation in the fat content of anchovy (Engraulis japonicus) in the Yellow Sea, and evaluated the potential effect of variation in fat content on the acoustic assessment of anchovy abundance. The fat content of anchovy varied seasonally, with two maxima and two minima in a year. The highest fat content (14.75%) was measured in the pre-spawning period in May, and the lowest fat content (2.48%) was measured during the post-spawning period in October. Fat content appeared to correlate with water content, but not body size. Assuming that the target strength is decreased by 0.2dB for every 1% increase in fat content, the seasonal difference in the target strength of anchovy may be as high as 2.45 dB. Given this, the acoustic abundance estimate may be biased by between 43% and 76%. Our results highlight the need for more information on the changes in fat content of fishes whose abundance is estimated by acoustic surveys.