State of the pacific oyster Crassostrea gigas (Thunberg, 1793) population in the Losos Bight (Sakhalin Island) in 2013–2020

The state of the aggregation of Pacific oyster Crassostrea gigas (Thunberg, 1793) in the Losos Bight (Aniva Bay) in 2013–2020 was assessed. For now, this aggregation has been in depression as a result of intense fishing. The abundance of the aggregation of commercial individuals (shell height >120 mm) was poor two recent years. The total stock estimated is 31.5 t; the commercial stock – 4.7 t.

The world’s tiniest land snails from Laos and Vietnam (Gastropoda, Pulmonata, Hypselostomatidae)

Two new, extremely small land snail species, Angustopila coprologos Páll-Gergely, Jochum & Hunyadi n. sp. and Angustopila psammion Páll-Gergely, Vermeulen & Anker n. sp. are described from northern Vietnam and northern Laos, respectively. The former is characterized by a rough surface sculpture and bears tiny mud granules arranged in a pattern of radial lines on its shell surface. The latter species is the new global record-holder of the tiniest land snail title, with a shell width of 0.6–0.68 mm and a shell height of 0.46–0.57 mm. These measurements surpass the former records of Angustopila pallgergelyi and Acmella nana.

Population structure and relative growth of rock snails Reishia bitubercularis (Gastropoda: Muricidae) from Ambon Island, Eastern Indonesia

The present study reports and discusses population structure and morphometric relationships of muricid <em>Reishia bitubercularis</em> in two populations from Ambon Island, Maluku. Field sampling was conducted at two locations with typical hard substrate i.e. Ambon Bay and Central Maluku, where each location comprises three different stations. Morphometric relationships were analyzed for shell dimensions i.e., shell length (SL), shell width (SW), shell height (SH) and total weight (TW). A total of 496 individuals of <em>R. bitubercularis </em>were collected in both locations. SL ranged from 7.07 to 42.38 mm in Ambon Bay, and 12.45 to 42.69 mm in Central Maluku. The highest number of individuals in Ambon Bay and Central Maluku was in size 26-28 mm and 30-32 mm, respectively. The mean SW/SL ratio ranged from 0.67 mm to 0.73 mm. Sex ratio was significantly different from 1:1, with females outnumbered males in both locations. Morphometric relationships indicated SL grows faster than SW in both locations and faster than SH and TW in Central Maluku. While the growth rate between SL vs SH; and TW vs SL are relatively similar in Ambon Bay. Overall, both Ambon Bay and Central Maluku have a similar variation of abiotic factors which also play an important role in shell morphometrics and relative growth of muricids, especially related to the exposed area in the rocky intertidal zone where sampling was conducted.

Impact of contamination on biology, biochemical, and histology of Eobania vermiculata and Monacha obstructa during different seasons in Ismailia, Egypt

Land snails are found to be an appropriate sentinel organism, and the biomarkers chosen are effective for terrestrial heavy metal biomonitoring. The study aimed to compare the biological, biochemistry, and histology of two land snails in the Ismailia governorate, Egypt during different seasons. Random snails were collected from January 2015 to December 2015 from two sites in the Ismailia Governorate, on wet days during each season's middle month. Soft tissues were taken from the dissected snails. It was noticed that most of the morphometric parameters measured shell height (ShH), last whorl width (LWW), maximum diameter (MaxD), aperture height (AH), and aperture width (AW) were higher in Eobania vermiculata (Sp. 1) than those measured in Monacha obstructa (Sp. 2), except for shell height measurement, which was the same in both species. The growth rate of Sp. 1 and Sp. 2 changed seasonally. In the more polluted areas with heavy metals, lipid peroxidation (LPO) was higher in snails and total protein content than in the snails collected from the less polluted areas for all seasons. However, the snails displayed lower levels of glutathione (GSH) as compared to snails at a less polluted site. GSH and LPO levels, on the other hand, have a negative relationship between them. Histopathological alterations in the digestive gland were more obvious in the general architecture of the digestive gland that had lost its tubular appearance. The excretory cells showed an increase in their excretory granules' number and size while calcium cells decreased. Also, gonad follicles have lost their normal architecture with the degeneration of some stages of spermatogenesis and oogenesis. In conclusion, There was a strong correlation between GSH levels and total protein content in the same soft tissues. GSH and LPO levels, on the other hand, have a negative relationship. The overall results display the usefulness of E. vermiculata and M. obstructa land snails as bioindicator organisms and support the application of this ecotoxicological approach for evaluating the biologic impact of toxins. E. vermiculata is more abundant than M. obstructa . The density, morphometric, biochemical, and histology of E. vermiculata and M. obstructa were different at different seasons.

INTERNAL QUALITIES OF EGG(S) OF LAYING BIRDS FED DIETS CONTAINING VERNONIAAMYGDALINALEAF (BITTER LEAF) MEAL AT DIFFERENT GROWING PHASES

One hundred and twenty eight Isa-Brown pullet chicks were used for this study to determine the internal qualities of egg(s) from birds fed diets containing Vernoniaamygdalina leaf meals at different growing phases. The leaves used were air-dried, crushed and milled, and was then incorporated into the experimental treatments (T2, T3, and T4) at 2% inclusion each. The birds were randomly allotted into four dietary treatments (T1, T2, T3 and T4) and each treatment was replicated four times with each replicate having 8 birds from the chick phase to the growing phase, thereafter 6 birds per replicate were randomly picked at the laying phase (point of cage), with T1 serving as the control treatment throughout the experimental period of 39weeks. The parameters measured include egg weight, shell weight, shell thickness, shell height, yolk weight, yolk height, yolk width, albumin length, albumin height, albumin weight, while yolk index and Haugh unit were calculated. The result of the experiment showed that T3 with 2% inclusion of Vernoniaamygdalinafrom the growing phase had the best shell thickness, yolk color and acceptable value for Haugh unit. Other parameters measured showed that yolk weight, and Haugh unit were significant at (p<0.05) with 2% inclusion of Vernoniaamygdalina. Every other parameter measured showed no significant difference across the treatments at (p>0.05). The result showed that 2% inclusion of Vernoniaamygdalinaleaf meal at the growing phase of the experiment improved the internal qualities of eggs.

Molecular Characterization and Expression of the LAP3 Gene and Its Association with Growth Traits in the Blood Clam Tegillarca granosa

Leucine aminopeptidase 3 (LAP3) is a metallopeptidase that cleaves N-terminal residues and is involved in protein maturation and degradation. In this study, we characterized the leucine aminopeptidase 3 (LAP3) gene from Tegillarca granosa (Tg-LAP3 for short), which appeared to consist of 15,731 nucleotides encoding 530 amino acids. We identified 12 introns and 13 exons in the Tg-LAP3 gene, suggesting a highly conserved genomic structure. The proximal promoter sequence consists of 1922 bps with a typical TATA box structure, which is the general structural characteristic of core promoters in eukaryotes. We found two functional domains in the Tg-LAP3 protein, including an N-terminal domain (41–174aa) and a peptidase_M17 catalytic domain (209–522aa). Multiple alignment showed that Tg-LAP3 shares 73.4% identity with LAP3 of Mizuhopecten yessoensis and 55.2–70.7% identity with LAP3 of other species. Quantitative analysis of Tg-LAP3 in embryos/larvae and adult tissues indicated that the highest expression occurred in eyebot larva, with limited expression in other stages; among tissues, the highest expression was found in the liver (p < 0.05). Association analysis found that three single-nucleotide polymorphisms (SNPs) (g.-488A > G, g.-1123C > T, and g.-1304C > A) in the proximal promoter were successfully typed, but there was no significant difference in growth traits (body weight, shell length, shell width, and shell height) among these genotypes. The results of our study demonstrate the functional roles of the Tg-LAP3 gene and provide valuable information for molecular marker-assisted selection (MAS) of the blood clam.

The methylome of Biomphalaria glabrata and other mollusks: enduring modification of epigenetic landscape and phenotypic traits by a new DNA methylation inhibitor

Background 5-Methylcytosine (5mC) is an important epigenetic mark in eukaryotes. Little information about its role exists for invertebrates. To investigate the contribution of 5mC to phenotypic variation in invertebrates, alteration of methylation patterns needs to be produced. Here, we apply new non-nucleoside DNA methyltransferase inhibitors (DNMTi) to introduce aleatory changes into the methylome of mollusk species. Results Flavanone inhibitor Flv1 was efficient in reducing 5mC in the freshwater snails Biomphalaria glabrata and Physa acuta, and to a lesser degree, probably due to lower stability in sea water, in the oyster Crassostrea gigas. Flv1 has no toxic effects and significantly decreased the 5mC level in the treated B. glabrata and in its offspring. Drug treatment triggers significant variation in the shell height in both generations. A reduced representation bisulfite-sequencing method called epiGBS corroborates hypomethylation effect of Flv1 in both B. glabrata generations and identifies seven Differential Methylated Regions (DMR) out of 32 found both in Flv1-exposed snails and its progeny, from which 5 were hypomethylated, demonstrating a multigenerational effect. By targeted bisulfite sequencing, we confirmed hypomethylation in a locus and show that it is associated with reduced gene expression. Conclusions Flv1 is a new and efficient DNMTi that can be used to induce transient and heritable modifications of the epigenetic landscape and phenotypic traits in mollusks, a phylum of the invertebrates in which epigenetics is understudied.

Growth and survival of the hatchery juveniles of pacific oyster Crassostrea gigas (Thunberg, 1793) in Peter the Great Bay

Growth and survival rates of juveniles of pacific oyster Crassostrea gigas are estimated for the first time in conditions of Peter the Great Bay (Japan Sea). The data were collected at artificial hatchery in the Aquaculture Center located on Popov Island in 2019. The juveniles were settled and reared on two types of substrate: 1) perforated plastic plates with the diameter of 30 cm (area 7.1 dm2), and 2) scallop shells with the height about 10 cm (average area 0.8 dm2) mounted in the western Peter the Great Bay (Voevoda Bay) in September 2019. Before placing in this site, the average height of the juveniles’ shells did not exceed 7 mm. In July 2020, after the 9-month exposure of the collectors with spat in the Voevoda Bay, the height increased in 12–18 times and reached 55–90 mm on the substrate 1 and 25–65 mm on the substrate 2. Then the oysters reared on the substrate 1 were replaced from the plastic plates to the cage shelves and the cages were moved to the Stark Strait. Difference in the growth rate became more apparent in September 2020, when the height of shells settled on the plastic plates reached 67 mm, on average (73 % in the range 50–85 mm) and for the shells settled on the scallops — 32 mm, on average (72 % in the range 20–45 mm). The shell height increased to 76.6 and 52.4 mm, respectively, to the end of October 2020. Finally, about 59 % of the oysters reared on the substrate 1 and cage shelves and 9 % of the oysters reared on the substrate 2 reached the commercial size (80–100 mm). Their survival in the period from mid-October 2019 to July 2020 is estimated as 46.9 % (28.8–98.2 %) for the substrate 1 and 33.5 % (4.0–78.3 %) for the substrate 2. On the substrate 1, survival of the mollusks attached to lower surface of the plates was twice higher. The survival had a tendency to decrease with increasing of the juveniles density. The experiment showed good viability for artificially hatched juveniles of pacific oyster in conditions of Peter the Great Bay. Their growth in the Voevoda Bay and the Stark Strait, with necessary technological measures, as replacing, thinning, and substrate cleaning from fouling, allows to produce oysters with the commercial size.

Dynamics of stock for yesso scallop Mizuhopecten yessoensis (Jay, 1856) in the northwestern Tatar Strait from the beginning of its fishery to nowadays

Settlements of yesso scallop Mizuhopecten yessoensis in the northwestern Tatar Strait (within the waters of Khabarovsk Region) are considered on the data of scuba (< 20 m depth) and dredge (> 20 m depth) surveys conducted in 2001–2018. To 2018, only two settlements remain in this area from 9–12 ones in 2010–2014; both remained settlements have no commercial value. Trend to decreasing of the settlements density is shown: the mean density was 3.0 ind./m2 in 2001, 0.20 ind./m2 in 2010, and 0.0005 ind./m2 in 2018. Commercially valuable scallops with the shell height > 120 mm prevailed in the settlements (86–100 %), whereas juveniles were rare or absent (1.6 %, on average) in all years of surveys. CPUE decreased from 200 kg/diver/hour in 2001–2003 to 10 kg/diver/hour in 2018. These changes are obviously reasoned by overexploitation of the population using dredging and scuba gathering. Total commercial stock of yesso scallop dropped to the minimal value in 2018: 200 t, that was in 4.5 times lower than the established limit. Thus, 5 years ban is established for M. yessoensis landing in the area.

Optimizing Harvest Time Through Absolute and Relative Growth of the Black Clam, Chionista fluctifraga, Cultivated in the Intertidal Along the Southeastern Coast of the Gulf of California

The shell height-body weight relationship of the black clam Chionista fluctifraga cultivated in the intertidal of the southeastern Gulf of California, was evaluated for the first time, to improve the knowledge in determining its harvesting time. Clam seeds (n=900,000; 6.1±1.9 mm) were produced in the laboratory. The culture was divided into pre-fattening in racks on the bottom (2 months) and fattening directly in the bottom (16 months) from May 2018 to October 2019. Each month, 60 (n=1080) clams were randomly selected to stablish the morphometric relationship between shell height (SH) and body weight (BW). The growth rate (mm/d and g/d) was recorded. The SH-BW Interaction in both cultivation phases showed positive allometry. SH in prefattening and fattening registered a growth rate of 0.072 and 0.058 mm/d, respectively. In fattening, the interaction BW-SH displayed a high coefficient of determination (R2=0.99). A final survival rate of 90% was obtained. The results yielded the equations that describe the relative growth of C. fluctifraga for the fattening stage. A harvesting time after 10 months of fattening in the intertidal zone is stablished to reach the commercial size (35 mm, SH).