Cascading effects of the flathead grey mullet Mugil cephalus in freshwater eutrophic microcosmos

Flathead grey mullet (Mugil cephalus L.) stocked in fish ponds were long considered to feed primarily on detritus. However, recent research has found that they obtain much of their food from plankton and that they have a detrimental effect on pond zooplankton and large phytoplankton, whilst enhancing small phytoplankton. It has been suggested that flathead grey mullet may also increase the internal phosphorus loading of the ecosystem, which would also increase phytoplankton density. To test whether zooplankton removal or nutrient release from the sediment is the better explanation for phytoplankton enhancement in the presence of flathead grey mullet, the ecosystems of fish-less tanks, tanks with a 60 m mesh filter and tanks stocked at a fish density of 243 g m^-3 were compared. In the presence of flathead grey mullets, cladocerans, ostracods and chironomid larvae became scarcer than in the control tanks, while there were more small phytoplankton and mud snails. The green algae Cladophora sp. did not occur at all. The presence of a mechanical filter also reduced cladoceran, ostracod and chironomid densities and increased phtyoplankton and mud snail density. However, the situation observed in filter tanks was intermediate between that observed in the fish tanks and the control tanks, due to the lower filtering efficiency of the mechanical filter. The organic matter content of the sediment decreased throughout the experiment in the control and filter tanks, but remained stable in fish tanks. Phosphorus and nitrogen concentrations were not affected by any treatment. These results showed that flathead grey mullet enhanced phytoplankton development due to the removal of large cladocerans and not as a consequence of nutrient release. Furthermore, the flathead grey mullet strongly modified the benthic community, probably due to direct predation.     

Food items of the grey mullet Mugil cephalus in the Banc d'Arguin area (Mauritania)

The stomach and gut content of 22 Mugil cephalus from the Banc d'Arguin shallows (Mauritanian coast, West Africa) is examined. Quartz grains are the predominant material found in the stomach together with small portions of benthic diatoms and flakes, aggregates of fine-grained inorganic and organic particles. As the composition of this material is modified by a sorting procedure during feeding it is difficult to conclude, whether seagrass stands (muddy and mixed sediments with sand fraction) or sandy flats are preferred as feeding habitats. The animals studied did not or not relevantly utilize the non-diatom microphytes, though the environment of the Banc d'Arguin is rich in microbial mats (cyanobacteria), Vaucheria beds and epiphytic vegetation of seagrass leaves.     

Isolation and characterization of visceral excitatory neuropeptides from striped mullet (Mugil cephalus L.) brain

From a brain extract of the catadromous fish, striped mullet (Mugil cephalus), two visceral excitatory neuropeptides (Mvp-1 and Mvp-2) were isolated by means of reversed phase chromatography together with bioassay on fish hindgut. The primary structure of Mvp-1 was elucidated to be SGPAGVLamide by ESI-Q-TOF mass spectrometry. The threshold concentration of Mvp-1 that changes spontaneous contraction of mullet hindgut was between 10(-9) and 10(-8) M. In addition, Mvp-1 was found to exert excitatory activities on some other smooth muscle segments (oviduct and esophagus) of mullet but it did not show any effect on body wall muscle strips. Therefore, the present study suggests that Mvp-1 and Mvp-2 peptides act as factors that control visceral contractions of mullet gastrointestinal tract.     

鲻鱼甲状腺免疫组织化学与超微结构

用组织学、免疫组织化学和电镜技术对鲻鱼甲状腺的分布与形态结构进行研究。结果表明，鲻鱼甲状腺滤泡分散地分布在第1对至第3对鲻弓的入鳃动脉之间；构成同一滤泡的单层上皮细胞呈扁平形和立方形两种形态，反映出同一滤泡中上皮细胞生理活动的不同步，且发现不同发育时期的鲻鱼这两种形态细胞的比例有显著差异。用特异性甲状腺球蛋白(TG)抗体对甲状腺滤泡的免疫染色反应显示，阳性物质分布在滤泡上皮细胞的脑膜及围绕滤泡腔的周边与中央胶质，幼年与成年鲻鱼甲状腺滤泡的免疫阳性反应部位有所不同。两种不同活动状态的细胞，扁平形细胞和立方形细胞，在核与胞质的超微结构上有显著不同。扁平形细胞的核为扁平形或椭圆形，核内常染色质十分丰富，核膜光滑，核周腔和核孔没有扩张，胞质中各种细胞器发育较差；立方形细胞的核膜高度凹陷，将核分为2叶至3叶，核周腔和核孔扩张，胞质中各种细胞器发育良好，出现长棒形和哑铃形线粒体，且数量明显多于扁平形细胞，多层粗面内质网呈板层状排列，还有发育好的高尔基复合体。另外，我们还观察到一种介于扁平形细胞和立方形细胞之间的中间型细胞，其胞质和核的发育明显好于扁平形细胞，但比立方形细胞差。上述结果可为了解鲻鱼甲状腺滤泡细胞分泌活动的机制提供基础资料[动物学报49(2)：230—237，2003]。     

Population genetic structure of striped mullet, Mugil cephalus, along the coast of China, inferred by AFLP fingerprinting

The striped mullet, Mugil cephalus, is an economically important species for both aquaculture and commercial fisheries in China. In this study, the amplified fragment length polymorphism (AFLP) technique was employed to analyze population genetic diversity and genetic distance between different populations with the aim of elucidating the population structure and gene flow of M. cephalus along the coast of China. A total of 230 fragments with 150–500 bp were identified from 118 individuals by five AFLP primer combinations. The polymorphic loci within populations varied from 46.52% to 64.78%, with an average of 53.91%, and the average heterozygosity from 0.1829 to 0.2282, with an average of 0.2074. The UPGMA phenograms of 118 individuals were constructed based on Dice similarity coefficients and four clusters were recognized. AMOVA analysis revealed that 60.7% of genetic variations were within populations and 39.3% between populations. The estimated genetic distance (φ_ST) value over all polymorphic loci across the six populations was 0.393 (p < 0.0010), indicating a strong population structuring. The pairwise φ_ST value ranged from 0.1112 to 0.5358, with an average of 0.3693. The population pairwise gene flows (average N_m = 0.73) are low. In addition, the result of the Mantel test showed that there was a significant correlation between geographic and genetic distances (r = 0.5434, p = 0.0050). It was speculated that there exist at least four distinct geographic populational subdivisions of M. cephalus along the Chinese coast. This research has provided new molecular data which will aid our understanding of the genetic structure of this species.     

Isolation and charaterization of polymorphic microsatellite loci from so-iuy mullet (Mugil soiuy Basilewsky 1855)

The first set of polymorphic microsatellite markers were developed from so-iuy mullet (Mugil soiuy Basilewsky 1855). From a (GT)n-enriched genomic library, 53 microsatellites were selected for designing microsatellite primers, of which 36 gave working primer pairs. Ten of these loci were polymorphic in a test population of 24 individuals with alleles ranging from 3 to 9, and observed and expected heterozygosities from 0.2083 to 0.9167 and from 0.2651 to 0.8812, respectively. No significant linkage disequilibrium between pairs of loci was found, but two loci significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction. These polymorphic microsatellite loci should provide sufficient level of genetic diversity to investigate the fine-scale population structure and evaluate the breeding strategy in Mullet. Genbo Xu and Changwei Shao Contributed equally.     

A GM2-specific beta-hexosaminidase from the roe of striped mullet (Mugil cephalus)

The roe of striped mullet (Mugil cephalus) was found to contain a beta-hexosaminidase different from the beta-hexosaminidases isolated from other sources. The enzyme from mullet roe is able to cleave GalNAc from GM2 without the assistance of either an activator protein or a detergent. It also cleaves the oligosaccharide derived from GM2 and other oligosaccharides containing the GM2 sequence GalNAc beta 4(NeuAc alpha 3)Gal-. However, it is not effective in hydrolyzing neutral glycosphingolipids containing terminal GalNAc or GlcNAc, such as GbOse4Cer, GgOse3Cer, or LcOse3Cer. These results indicate that mullet roe beta-hexosaminidase can specifically cleave GalNAc from the glycoconjugates containing the GM2 sequence. No beta-hexosaminidase with such specificity has been previously described. Thus, this unique enzyme should be very useful for the detection and analysis of glycoconjugates containing the oligosaccharide chains with GM2 sequence.     

Effect of storage time on haematological parameters in mullet,Mugil cephalus

The aim of the present study was to assess the effect of storage time at +4 °C on red blood cell count (RBC), haematocrit (Hct), haemoglobin (Hb), white blood cell count (WBC), thrombocyte count (TC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) in mullet (Mugil cephalus) using an automatic method. After blood collection (T₀), all samples were analyzed using both the manual and automatic method. To test the validation of the automatic method, a paired t‐test was applied, and no statistical difference was observed. The samples were successively divided into four different aliquots and stored at +4 °C to assess the haematological parameters using the automatic method. The first aliquot was refrigerated for 6 h, the second one for 24 h, the third one for 48 h and the last one at for 72 h. One‐way repeated‐measures ANOVA showed a significant effect of storage time (P < 0.05) on Hb, WBC, TC, MCH and MCHC. These results suggest that haematological parameters can be assessed within 6 h from blood collection when samples are stored at +4 °C because long‐term storage modifies the results of the analyses. Further studies on these parameters could be still needed in various fish species to validate an appropriate method for haematological analysis useful not only for the evaluation of the health status of animal living in captivity and in aquaculture but also to have reliability environmental haematological biomarkers. Copyright © 2012 John Wiley & Sons, Ltd.     

Gonadal development and steroid hormone profile of wild caught grey mullet (Mugil cephalus)

The grey mullet Mugil cephalus is one of the popular and fast growing fishes being cultured in tropical and subtropical regions. In the present study, histological observation of gonadal development and corresponding changes in sex steroid levels from different maturity stages of wild caught male and female were studied. In female, testosterone and 17β-estradiol increased with the advancement of maturation and reached peak (17β-estradiol, 323 ± 13 pg/ml; testosterone, 938 ± 7.87 pg/ml) in mature stage, whereas the level of progesterone was maximum (488 ± 4.9 pg/ml) during ripe stage. Vitellogenin level in serum showed a similar trend as 17β-estradiol. In case of male, the testosterone level in serum increased gradually with advancement of maturation and was maximum (1820 ± 40.25 pg/ml) during ripe stage, whereas significant decrease in 17β-estradiol and progesterone was noticed with advancement of maturation. The fundamental information from this investigation would be useful for developing protocol for accelerating maturation and spawning under captive condition.     

Morphologic and Genetic Evidence for Mixed Infection with Two Myxobolus Species (Myxozoa: Myxobolidae) in Gray Mullets,Mugil cephalus,from Korean Waters

The present study was performed to trace the decisive evidence for mixed infection of 2 Myxobolus species, M. episquamalis and Myxobolus sp., in the gray mullet, Mugil cephalus, from Korean waters. Mullets with whitish cyst-like plasmodia on their scales were collected near a sewage plant in Yeosu, southern part of Korea, in 2009. The cysts were mainly located on scales and also found in the intestine. The spores from scales were oval in a frontal view, tapering anteriorly to a blunt apex, and measured 7.2 µm (5.8-8.0) in length and 5.3 µm (4.7-6.1) in width. Two polar capsules were pyriform and extended over the anterior half of the spore, measuring 3.5 µm (2.3-4.8) in length and 2.0 µm (1.5-2.2) in width. In contrast, the spores from the intestine were ellipsoidal, 10.4 µm (9.0-11.9) in length and 8.4 µm (7.3-10.1) in width. The polar capsules were pyriform but did not extend over the anterior half of the spore, 3.7 µm (2.5-4.5) in length and 2.2 µm (1.8-2.9) in width. The nucleotide sequences of the 18S rDNA gene of the 2 myxosporean spores from scales and intestine showed 88.1% identity to each other and 100% identity with M. episquamalis and 94.5% identity with M. spinacurvatura from mullet, respectively. By the above findings, it is first confirmed that mullets from the Korean water are infected with 2 myxosporean species, M. episquamalis and Myxobolus sp.     

Identification of the sex‐determining region in flathead grey mullet (Mugil cephalus)

Elucidation of the sex‐determination mechanism in flathead grey mullet (Mugil cephalus) is required to exploit its economic potential by production of genetically determined monosex populations and application of hormonal treatment to parents rather than to the marketed progeny. Our objective was to construct a first‐generation linkage map of the M. cephalus in order to identify the sex‐determining region and sex‐determination system. Deep‐sequencing data of a single male was assembled and aligned to the genome of Nile tilapia (Oreochromis niloticus). A total 245 M. cephalus microsatellite markers were designed, spanning the syntenic tilapia genome assembly at intervals of 10 Mb. In the mapping family of full‐sib progeny, 156 segregating markers were used to construct a first‐generation linkage map of 24 linkage groups (LGs), corresponding to the number of chromosomes. The linkage map spanned approximately 1200 cM with an average inter‐marker distance of 10.6 cM. Markers segregating on LG9 in two independent mapping families showed nearly complete concordance with gender (R² = 0.95). The sex determining locus was fine mapped within an interval of 8.6 cM on LG9. The sex of offspring was determined only by the alleles transmitted from the father, thus indicating an XY sex‐determination system.     

Isolation and characterization of microsatellite loci in the striped mullet,Mugil cephalus

Forty‐three microsatellites were isolated from grey mullet (Mugil cephalus) of Mediterranean origin using the standard enrichment methods followed by enhanced chemiluminescence for the rapid identification of clones containing microsatellites. Eleven microsatellites were characterized to assess genetic diversity of different populations surrounding the island of Sardinia in Italy. The versatility of these markers was also assessed using an Australian population. All 11 microsatellites were highly polymorphic with a mean heterozygosity of 0.834. The number of alleles ranged from 14 to 38 in the Sardinian populations and from 11 to 25 in the Australian samples.     

Effect of rearing density on the blood and tissues of mullet (Mugil cephalus L.)

This study evaluated the biological effects of high stocking density on mullet (Mugil cephalus). Fifty fish, caught from Faro lake, were divided into two groups, a low-density control group (n = 15), stocked at 3.4 kg/m³ in an 800 l tank and a high-density experimental group (n = 35), stocked at 8.4 kg/m³ an 800 l tank. The ammonia concentration in the control group tank reached 0.3 mg/l after 2 days while that in the experimental tank reached 5 mg/l. Haematological parameters and serum protein profiles were determined following blood sampling. An Unpaired T-test showed significant differences between the two groups on Red Blood Cell Count, Haematocrit, Haemoglobin, White Blood Cell Count, Mean Corpuscular Haemoglobin, Mean Corpuscular Haemoglobin Concentration, total proteins, albumin and α-globulins. In the experimental group, the disappearance of prealbumin and the γ-globulin fraction was particularly evident. Histopathological changes indicated that mucosal electrolytic failure was the main function compromised. This could eventually lead to death. The findings suggest that mullet is sensitive to stocking density linked to ammonia toxicosis.     

Respiratory responses of the striped mullet,Mugil cephalus (L.) to hypoxic conditions*

Ventilation volume, ventilatory frequency, ventilatory stroke volume, percentage utilization of oxygen and respiratory metabolism were measured on unanaesthetized striped mullet, Mugil cephalus L., under ambient and hypoxic conditions with a modified van Dam respiration chamber. Hypoxia caused a statistically significant increase in ventilation volume, ventilatory frequency, and ventilatory stroke volume and a significant decrease in percentage utilization of oxygen. The routine rate of respiratory metabolism was not significantly altered by hypoxia. These responses probably represent ventilatory adjustments which serve to maintain a constant oxygen supply to the gills under conditions of oxygen depletion.     

Neuroendocrine regulation of puberty in fish: insights from the grey mullet (Mugil cephalus) model

We investigated the molecular regulation of pubertal development in the grey mullet, Mugil cephalus, a relatively late-maturing teleost fish. We have isolated and characterized the cDNAs of key reproductive genes along the brain-pituitary-gonadal (BPG) axis as well as the promoters of genes that modulate the axis at multiple levels. Together with relevant findings from other model species, we propose a conceptual model of the neuroendocrine regulation of puberty in the female grey mullet. Research areas that warrant further investigation are identified in the model.     

Pronounced genetic structure of mitochondrial DNA among populations of the circumglobally distributed grey mullet (Mugil cephalus)

Mitochondrial (ml) DNA genotypes of grey mullet ( Mugil cephalus ) from ocean basins around the world were analysed to estimate the amount of genetic differentiation in this cosmopolitan but mostly coastally-restricted species. Extensive genetic diversity was observed. Among 115 specimens from nine locales, 26 different haplotypes were detected using a battery of 13 restriction endonucleases. In phenetic analyses, these haplotypes grouped into seven distinct clusters whose members were in almost perfect accord with the geographic sources of the samples. Thus contemporary gene flow between the widespread collection locales must be absent or nearly so. Results contrast with the relative uniformity in mtDNA composition previously reported for populations of some circumglobally distributed pelagic fishes, and demonstrate that certain marine fishes with cosmopolitan distributions can exhibit pronounced population genetic structure even in the face of morphological conservatism.