Hematological characteristics of the spotted rose snapper Lutjanus guttatus (Steindachner, 1869) healthy and naturally infected by dactylogyrid monogeneans

The aim of the present study was to obtain a basic knowledge of the hematology in order to determine changes in blood parameters of the spotted rose snapper Lutjanus guttatus. The morphological features of blood cells were described according to the observations made by light microscopy of Wright-Giemsa-stained blood films. The reference intervals and the mean value were determined for each hematological parameter evaluated in healthy fish and data were compared to those of naturally infected, with dactylogyrid monogeneans fish.Infected fish showed a prevalence of 100% and a mean intensity of 246.6 parasites per fish. Mean values of HCT, WBC, thrombocytes percentage and eosinophils percentage were significantly higher (P < 0.05) in the infected fish. In addition, lymphocytes percentage and total protein were significantly lower (P < 0.05) in the infected fish compared to healthy fish. Only total WBC count, lymphocytes percentage and eosinophils percentage in infected fish were outside reference interval. The hematology of the spotted rose snapper of this study might serve as a basis for future studies and diagnosis. Changes observed in blood parameters in infected fish suggest that the immune system of L. guttatus was affected by the presence of the parasites.     

Expression and activity of trypsin and pepsin during larval development of the spotted rose snapper Lutjanus guttatus

The present study aimed to describe and understand the development of the digestive system in spotted rose snapper (Lutjanus guttatus) larvae from hatching to 40 days post-hatch (dph). The mouth opened between 2 and 3 dph, at that moment the digestive tract was barely differentiated into the anterior and posterior intestine, although the liver and pancreas were already present. Gastric glands were observed until 20 dph, followed by the differentiation of the stomach between 20 and 25 dph. Trypsinogen expression and trypsin activity were detected at hatching, increasing concomitantly to larval development and the change in the type of food. Maximum levels of trypsinogen expression were observed at 25 dph, when animals were fed with Artemia nauplii, and maximum trypsin activity was detected at 35 dph, when larvae were fed with an artificial diet. On the other hand, pepsinogen gene expression was detected at 18 dph, two days before pepsin enzymatic activity and appearance of gastric glands. Maximum pepsin activity was also observed at 35 dph. These results suggest that in this species weaning could be initiated at an earlier age than is currently practiced (between 28 and 30 dph), since larvae of spotted rose snapper develop a functional stomach between days 20 and 25 post-hatch.     

Post-transcriptional silencing of myostatin-1 in the spotted rose snapper (Lutjanus guttatus) promotes muscle hypertrophy

Molecular Biology Reports - Muscle growth is regulated by several factors including the growth differentiation factor 8, known as myostatin, an inhibitor of myocyte differentiation and...     

Expression of myostatin in the spotted rose snapper Lutjanus guttatus during larval and juvenile development under cultured conditions

In this study, the developmental expression pattern of myostatin (mstn) in the spotted rose snapper Lutjanus guttatus under culture conditions is presented. The full coding sequence of mstn from L. guttatus was isolated from muscle tissue, obtaining 1134 nucleotides which encode a peptide of 377 amino acids. The phylogenetic analysis indicated that this sequence corresponds to mstn‐1. mstn expression was detected in embryonic stages, and maintained at low levels until 28 days post‐hatch, when it showed a significant increase, coinciding with the onset of metamorphosis. After that, expression was fluctuating, coinciding probably with periods of rapid and slow muscle growth or individual growth rates. mstn expression was also analysed by body mass with higher levels detected in smaller animals, irrespective of age. mstn was also expressed in other tissues from L. guttatus, presenting higher levels in brain, eye and gill. In brain for instance, two variants of mstn were isolated, both coding sequences were identical to muscle, except that one of them contained a 75 nucleotide deletion in exon 1, maintaining the reading frame but deleting two conserved cysteine residues. Phylogenetic analysis indicated that this brain variant was also mstn‐1. The function of this variant is not clear and needs further investigation. These results indicate that mstn‐1 participates in different physiological processes other than muscle growth in fishes.     

A new species of Euryhaliotrematoides Plaisance & Kritsky, 2004 (Monogenea: Dactylogyridae) from the gills of the spotted rose snapper Lutjanus guttatus (Steindachner) (Perciformes: Lutjanidae)

Euryhaliotrematoides mehen n. sp. (Monogenea: Dactylogyridae) is described based on specimens collected from the gills of the spotted rose snapper Lutjanus guttatus (Steindachner) off Mazatlan, Sinaloa, on the northwestern coast of Mexico. The synapomorphy of this genus is the presence of a funnel-shaped base of the male copulatory organ. This new species differs from all other species of the genus by possessing a male copulatory organ with a base with a thickened margin and a membranous accessory piece resembling a scarf and covering about 60% of its distal region. PCR products of the 28S rRNA (831 bp) and 18S rRNA (662 bp) genes were sequenced and submitted to GenBank (accession numbers HQ615997 and JF938069, respectively). BLASTn searches revealed no 100% identical hits with the previously registered monogenean sequences.     

Effects of feed restriction on growth performance,lipid mobilization,and gene expression in rose spotted snapper (Lutjanus guttatus)

The effects of feed deprivation were evaluated for 1 week and 2 weeks in Lutjanus guttatus juveniles. A significant reduction in body mass was observed in both feed deprivation schemes, as well as in hepatosomatic, viscerosomatic and mesenteric fat indexes. The composition of fasted fish was characterized by a decrease in lipid content; the liver displayed an intense reduction of lipid reserves in both fasted groups, and increased expression of the lysosomal acid lipase. 1 week after re-feeding, both experimental groups showed an increase in specific growth rate, feed intake and feed conversion ratio. A recovery in hepatic lipid reserves was observed, and the expression of the lysosomal acid lipase decreased, although lipid content in both groups was still significantly lower than in control groups. Hepatic expression of the growth hormone receptor decreased after fasting, and remained low even after the fish were fed, whereas the expression of insulin-like growth factor 1 and 2 increased after fasting and decreased in both groups when fish were fed again. Altogether, these results showed a partial compensatory growth response in L. guttatus juveniles after fasting, with enhanced growth rates and improved feed efficiency. Fish used stored lipid reserves as the main energy source, and the expression of growth-related and lipid mobilization marker genes in the liver showed similar patterns in both fasting schemes. Based on the results, we suggest as much as 1-week fasting intervals during grow-out programmes to reduce visceral fat and increase growth rate in this species.

     

Diel spawning periodicity of red snapper Lutjanus campechanus in the northern Gulf of Mexico

Ovaries of red snapper Lutjanus campechanus were examined histologically to determine rates of oocyte maturation, diel spawning periodicity and whether lunar cycle influenced spawning rhythm. Hydration of red snapper oocytes began during the mid‐morning hours; c . 5 h was necessary for oocytes to become fully hydrated and ovulation occurred no more than 5 h after oocytes attained full hydration. Appearance of fresh postovulatory follicles after 1330 hours and the absence of hydrated oocytes after 1830 hours signified that red snapper spawning occurred during this 5 h period. In addition, evidence of a peak in spawning was seen near 1600 hours. Postovulatory follicles degenerated within a 24 h time period. A lunar spawning cycle was not evident.     

Isolation and characterization of microsatellite loci from mangrove red snapper Lutjanus argentimaculatus

Lutjanus argentimaculatus, also called mangrove red snapper, is a commercially important fish in East Asia. A proper understanding of population structure is primarily linked with the management of genetic resources in exploiting marine fisheries. Herein, seven microsatellite loci, which showed high polymorphism (observed heterozygosity per locus ranging from 0.3571 to 0.7857 and expected heterozygosity per locus ranging from 0.6236 to 0.8821), were isolated and characterized from L. argentimaculatus. Cross‐species amplifications also indicate that primers designed for these loci may be useful for further studies about other closely phylogenetic species of the family Lutjanidae.     

Parasite communities of the spotted rose snapper Lutjanus guttatus (Perciformes: Lutjanidae) off the Mexican Pacific coasts: Spatial and long-term inter-annual variations

A total of 802 individuals of Lutjanus guttatus (Steindachner, 1869) specimens were collected over a 10-year period (August 2012 to February 2021) from four locations on the south-central Pacific coast of Mexico. Their parasite communities were quantified and analyzed to determine if they experience significant spatial and inter-annual variations.Thirty-two taxa of metazoan parasite were recovered and identified: four species of Digenea, four Monogenea, one Cestoda, two Acanthocephala, seven Nematoda, one Hirudinea, and nine of Crustacea (six Copepoda, and three Isopoda). Species richness was greatest among the digenean group, which represented 25% of the total species recovered, followed by the nematodes (22% of total species). Species richness at the component community level (10 to 20 species) was similar to reported richness in other Lutjanus spp. The component communities and infracommunities exhibited a similar pattern: low species richness and diversity, and dominance by a single species, mainly the monogenean Haliotrematoides guttati. Parasite community structure and species composition varied through the years, as well as between sampling locations. Variations were possibly caused by a combination of biotic and abiotic factors which generated notable changes in the infection levels of several component species. However, the similarity in the parasite species composition was high locally for short-term periods (one or two years). This result, therefore, suggests that parasite communities of L. guttatus may be more predictable locally, but only for short-term periods.     

Protection of red snapper (Lutjanus sanguineus) against Vibrio alginolyticus with a DNA vaccine containing flagellin flaA gene

Aims: The main aims of this study were to construct a DNA vaccine containing flagellin flaA gene from Vibrio alginolyticus strain HY9901 and to explore the potential application of pcDNA‐flaA as a DNA vaccine candidate for red snapper (Lutjanus sanguineus). Methods and Results: Plasmid DNA encoding flagellin flaA gene (designated as pcDNA‐flaA) was used as a DNA vaccine to immunize red snapper. The distribution, expression and immunoprotection of the DNA vaccine were analysed in tissues of the red snapper by PCR, RT‐PCR and challenge test. PCR results indicated that pcDNA‐flaA distributed in liver, spleen, kidney, gill and injection site muscle at 7–28 days after vaccination. RT‐PCR results indicated that the flaA gene was expressed in all above tissues of vaccinated fish at 7–28 days after vaccination. In addition, fish receiving the DNA vaccine developed a protective response to live V. alginolyticus challenge 28 days post inoculation, the relative per cent survival (RPS) was 88%. Conclusions: This study showed that injection of pcDNA‐flaA induced an efficient, systemic and antigen‐specific immune response in red snapper, which makes it an effective vaccine candidate against V. alginolyticus infection. Significance and Impact of the Study: The finding that red snapper does adequately respond to pcDNA‐flaA intramuscular injection makes pcDNA‐flaA a promising candidate for DNA vaccine treatment. Furthermore, the availability of red snapper for foreign gene expression represents a useful model to develop effective prophylactic strategies and opens new perspectives for the treatment of bacterial pathogens of marine cultured fish.     

Isolation and characterization of microsatellite DNA loci from Russell's snapper (Lutjanus russellii)

A total of 37 microsatellite loci from the Russell's snapper, Lutjanus russellii, were successfully isolated and characterized. Thirty‐four loci were polymorphic in L. russellii samples. Twenty of the 37 markers did not differ significantly from Hardy–Weinberg expected genotype proportions. Significant linkage disequilibrium was detected in one pairwise comparison. The numbers of alleles and observed heterozygosities in polymorphic loci ranged from two to 16 and from 0.41 to 0.95, respectively. These markers will be useful for studying the population genetic structure of this species.     

Osmoregulatory capabilities of the gray snapper,Lutjanus griseus: salinity challenges and field observations

We investigated the osmoregulatory responses (plasma osmolality and blood hematocrit) displayed by the gray snapper 6–192?h after abrupt changes in ambient salinity. Fish were challenged with six different salinity treatments including a control (0, 5, 30, 50, 60, and 70?ppt) and blood samples were collected at various time points post-transfer. Gray snapper across all size classes tested (13.5–24.5?cm total length) acclimated successfully to hypo- and hyper-saline environments (0–60?ppt) after an adjustment period of ～96?h. However, abrupt transfers to 70?ppt resulted in 100% mortality within 48?h. Laboratory results were then compared with field measurements obtained after fish were captured in low salinity (0–4?ppt) or marine (～30?ppt) habitats, suggesting that osmoregulatory processes occurred similarly in both settings. Overall, findings suggest that gray snapper possess similar or higher osmoregulatory capabilities compared to many euryhaline species examined to date, and thus should be considered a euryhaline species.     

斑鳠烂鳃病病原菌的研究

从池塘患烂鳃病的斑鳠(Mystus guttatus)亲鱼病灶中分离出一株毒力较强的致病菌-Mg2,经形态学观察、生理生化特性和16S rRNA基因序列分析鉴定,其主要特征为：菌体细长,无鞭毛和荚膜,革兰氏阴性,大小0.5×(6.5－11)μm,菌落黄色,边缘不整齐呈假树根状。氧化酶、过氧化氢酶阳性,分解酪素和明胶,硝酸盐还原阳性;不分解纤维素、几丁质、酪氨酸、七叶灵和淀粉,吲哚和葡萄糖产气阴性。对恩诺沙星、诺氟沙星、萘啶酸、红霉素、洁霉素敏感,菌株的16S rRNA基因序列分析结果表明：Mg2菌株与柱状黄杆菌聚类,基因序列的同源性在97.5%以上,综合生理生化、分子生物学两方面的分类鉴定结果,Mg2菌株应归类鉴定为柱状黄杆菌（Flavobacterium Columnare）。     

Microsatellite Variation Among Red Snapper (Lutjanus campechanus) from the Gulf of Mexico

Allelic variation at a total of 20 nuclear-encoded microsatellites was examined among adult red snapper (Lutjanus campechanus) sampled from 4 offshore localities in the Gulf of Mexico. The number of alleles at the 20 microsatellites ranged from 5 to 20; average (± SE) direct count heterozygosity values ranged from 0.148 ± 0.025 to 0.902 ± 0.008. No significant departures from expectations of Hardy-Weinberg equilibrium were found for any locus within samples, and genotypes at pairs of microsatellites appeared to be randomly associated, i.e., in genotypic equilibrium. Tests of homogeneity in allele distributions among the 4 localities were nonsignificant for 19 of the microsatellites. Allele distribution at microsatellite Lca 43 was heterogeneous among localities before (but not after) Bonferroni corrections for multiple tests executed simultaneously. Tests of homogeneity in the distribution of individual alleles at Lca 43 gave similar results: one low frequency allele was distributed heterogeneously among samples before, but not after, Bonferroni correction. Molecular analysis of variance indicated that more than 99% of variation at each microsatellite was distributed within sample localities. These results generally are consistent with the hypothesis of a single population (stock) of red snapper in the northern Gulf of Mexico. Received September 25, 2000; accepted January 16, 2001     

Evidence for spatial limitation of the bluestripe snapper Lutjanus kasmira in French Polynesia from parasite and otolith shape analysis

The study focused on small‐scale location and movement patterns of the bluestripe snapper Lutjanus kasmira on the north coast of Moorea (Society Archipelago, French Polynesia, south‐central Pacific). Juveniles of this species occur in the estuary, and adults occur widely in the lagoon, the outer slope and in the intermediate channel, where fish aggregate in large schools during daytime. While fish were all sampled within a few hundred metres, they exhibited significantly different parasite fauna and otolith shapes according to their locality: estuary, lagoon, channel and outer slope. While juveniles did not exhibit any parasites in the estuary parasitological and otolith‐shape data suggest that adults rarely move between their three adjoining habitats, and more interestingly that large aggregations in the channel are formed predominantly by resident individuals with limited local movement, including at night. Besides giving some information on the host biology, such findings may have application in local fisheries management.     

Effects of exploitation on reproductive capacity of blackspot snapper, Lutjanus fulviflamma (Pisces: Lutjanidae) in Mafia Island, Tanzania

There is paucity of information on the effects of exploitation on reproductive characteristics of blackspot snapper, Lutjanus fulviflamma (Forsskål 1775) in Tanzanian coastal waters. We compared size at first sexual maturity (L_M50), sex ratio, fecundity, and breeding season of L. fulviflamma in least fished Mafia Island Marine Park (MIMP) and intensively fished areas (IFA) between May 1999 and April 2001. Fish in MIMP matured at significantly smaller size (female: L_M50 = 206.3 mm; male: L_M50 = 195.5 mm) than in IFA (female: L_M50 = 216.7 mm; male: L_M50 =212.1 mm) total body length. Sex ratio was balanced at 1.03 : 1 (female : male) in MIMP, but it was skewed 0.9 : 1 (female : male) in IFA. Size‐related differences in sex ratio were observed with males predominating in the smaller sizes and females in the larger sizes. Total fecundity of fish in MIMP was determined at 45,200–430,200 oocytes in females of between 207 and 293 mm total length. Lutjanus fulviflamma in MIMP has a prolonged spawning season lasting from September to March peaking in December. None of the fish from IFA were in breeding state, suggesting recruitment overfishing is an added matter of concern for the long‐term sustainability of the fishery at the current exploitation level.     

The food and feeding habits of blackspot snapper, Lutjanus fulviflamma (Pisces: Lutjanidae) in shallow waters of Mafia Island, Tanzania

Natural diet and feeding habits of blackspot snapper, Lutjanus fulviflamma (Forsskål 1775) were investigated in Mafia Island Marine Park (MIMP) and intensively fished areas (IFA) between May 1999 and April 2001. An index of relative importance (IRI) was used to assess the food preference for L. fulviflamma. Feeding habits were studied with respect to sex and size of the predator, seasons and feeding periodicity. Of 4642 L. fulviflamma specimens sampled, 46% had empty stomachs. Brachyurans were the main prey of L. fulviflamma accounting for 48% IRI. Benthic invertebrates dominated by other crustaceans, gastropods, bivalves, cephalopods and polychaetes and demersal teleosts made up the remainder of the diet. Female and male L. fulviflamma were not effectively segregated by trophic dimension in the study areas. Specimens in the 90–120 mm total length (TL) fed exclusively on crustaceans, with shrimps accounting for 40% IRI. Lutjanus fulviflamma in the 270–300 mm TL preferred fish prey accounting for 50% IRI. The diet of fish species showed clear spatial differences, which were dependent on size distributions. Lutjanus fulviflamma fed mostly but not exclusively at night peaking at dusk and dawn. The results clearly demonstrate that L. fulviflamma is an opportunistic forager, which exhibits both diel and ontogenetic feeding habits.     

Construction of a fusion flagellin complex and evaluation of the protective immunity of it in red snapper (Lutjanus sanguineus)

Aims: The main aims of this study were to construct a bivalent subunit vaccine containing flagellin flaA gene and flagellin flaB gene from Vibrio alginolyticus strain HY9901 and to explore the potential application of the fusion protein FlaA‐(G₄S)₃‐FlaB as a vaccine candidate for red snapper (Lutjanus sanguineus). Methods and Results: Flagellin gene flaA and flaB of V. alginolyticus were linked by gene SOEing (gene splicing by overlap extension) technology. The expression of the fusion gene flaA‐(G₄S)₃‐flaB in Escherichia coli BL21(DE3) was confirmed by SDS‐PAGE. Western blot analysis showed that the fusion protein FlaA‐(G₄S)₃‐FlaB, which was purified by affinity chromatography on Ni‐NTA resin, had positive reaction with mouse anti‐FlaA serum and mouse anti‐FlaB serum, respectively. The immunoprotection of FlaA‐(G₄S)₃‐FlaB as a bivalent subunit vaccine was investigated in red snapper model by enzyme‐linked immunosorbent assay (ELISA) and challenge test. Red snapper vaccinated with FlaA‐(G₄S)₃‐FlaB produced specific antibodies and were highly resistant to infection by virulent V. alginolyticus. Conclusions: The fusion gene flaA‐(G₄S)₃‐flaB from V. alginolyticus strain HY9901 was cloned by gene SOEing and was expressed in E. coli. This fusion protein FlaA‐(G₄S)₃‐FlaB is a good protective antigen of V. alginolyticus and should be considered as an effective vaccine candidate against infection by V. alginolyticus in red snapper. Significance and Impact of the Study: Two flagellin genes, flaA and flaB, which are independent in structure and function, were first linked together by gene SOEing technology. The finding that red snapper did adequately respond to the fusion protein FlaA‐(G₄S)₃‐FlaB injection made it a promising candidate for vaccine treatment. To develop effective vaccine candidates against V. alginolyticus, more attention should be given to these immunogenic flagellins.     

Morphological and biochemical effects of food deprivation during the early development of Pacific red snapper Lutjanus peru

We report the effects of food deprivation on the early development of Pacific red snapper Lutjanus peru during the first days of development. The point of no return (PNR) was determined using the feeding incidence after a delay in first feeding. The gradual deterioration of the larvae during food deprivation was recorded using morphometric, histological, enzymatic and biochemical analysis. The time to reach the PNR was 120 h after hatching. Morphologically, the total length, muscle height, head length, tail length and pectoral angle showed the biggest reductions and their growth coefficients changed significantly during food deprivation. Histologically, enterocyte height also was reduced significantly. The protein concentration and activities of the digestive enzymes trypsin, cathepsin-like and lipase showed a significant decrease; meanwhile, amylase activity remained constant during food deprivation. The concentration of total essential free amino acids (EFAAs) decreased significantly while that of the nonessential free amino acids (NEFAAs) remain stable during food deprivation. The most abundant EFAAs were lysine, leucine, isoleucine and valine; the most abundant NEFAAs were alanine, glycine and glutamate, suggesting a more prominent role as energy substrates. At the time of the PNR the concentration of almost all the free amino acids showed a significant decrease. Early food deprivation has a significant impact on the morphology and biochemical characteristics of L. peru. These results suggest that initial feeding of L. peru should begin within 3 days of yolk sac depletion to avoid the PNR. Further studies are necessary to confirm and validate the characters identified in this study as biomarkers of starvation under culture conditions and evaluate their possible utility in ichthyoplankton surveys.     

Within-shelter behaviour of the spotted spiny lobster, Panulirus guttatus (latreille), in simulated communal dens: an exploratory study

Panulirus guttatus is a sedentary spiny lobster that dwells in the coral reef habitat. We conducted an exploratory study to describe the within-shelter behaviour of groups of 4-6 individuals of P. guttatus in simulated communal dens deployed in tanks. Lobsters displayed a diel within-shelter activity rhythm, with a peak shortly before dawn due to an increase in agonistic interactions among sheltered individuals. In a single-chamber cave, aggressions resulted in the constant rearrangement of individuals in the den, related to their preference to cling to the walls. In a triple-chamber cave, the largest male often defended one chamber, but the remaining individuals tended to aggregate in one of the other two chambers during the day. These trends suggest that P. guttatus is a gregarious lobster, but that this gregariousness is influenced by the amount of available shelters and by temporary, size-related social hierarchies. This hypothesis should be tested with fully replicated experiments to help understand the patterns of shelter utilization of P. guttatus in the coral reef habitat.