Morphofunctional changes of the digestive gland in the bivalve mollusk Crenomytilus grayanus (Dunker, 1853) in normal conditions and after parasitic invasion by trematodes

Specifics of the digestive cycle were studied in normal individuals of Gray’s mussel Crenomytilus grayanus and in those infested by trematodes from an unpolluted area of the sublittoral zone of Peter the Great Bay in May (during the day), June, July, and August. Four types of tubules corresponding to four phases of the digestive cycle were identified, and three varieties of destructing tubules were distinguished. It was shown that normally phases of absorption and digestion prevailed in the digestive gland. The portion of absorptive tubules (type II) was 95% in spring and 79% in summer. The total portion of destructing and restoring tubules (types III and IV) did not exceed 10% in spring and 20% in summer, and portion of tubules at the initial condition (type I) was 4% in spring and 2% in summer. During the day, insignificant variations in the ratio of different type tubules were recorded. It was revealed that, normally, granulocytomes were formed during utilization of necrotic tubules. Parasitic invasion of trematodes entailed swelling of the gland and a change in the proportion of tubules: the number of absorptive tubules decreased to 20%, the number of destructing and restoring tubules increased up to 60% and 12%, respectively.     

Histopathology of the digestive gland of the bivalve mollusk Crenomytilus grayanus (Dunker, 1853) from southwestern Peter the Great Bay, Sea of Japan

The histomorphology of the digestive gland of the bivalve mollusk Crenomytilus grayanus from Sivuchya Bay, which is located in the southwest of Peter the Great Bay and subjected to the effect of polluted waters of Tumannaya River, was studied. Pathological changes of the digestive tubules, channels, and connective tissue of the gland were recorded in all the mussels studied. The epithelium of the tubules and channels was characteristic with erosive disturbances and by heavy vacuolization of digestive cells; connective tissue of the gland was specified by cells with lipofuscin (granulocytomes) and by foci of cells necrosis and lysis. Nervous fibers running in the gland were swollen in some mollusks. Strongly basophilic spherical formations, presumably one of the development stages of a parasitic plasmodium, were found in the granulocytomes and among vesicular cells of connective tissue of all the mussels. It was concluded that pathological changes in digestive gland of Gray’s mussel might be caused by chronic pollution of the bay and by parasitic invasion.     

Neuroplastic and neuropathological changes in the central nervous system of the Gray mussel Crenomytilus grayanus (Dunker) under environmental stress

We studied here neuron ultrastructure, synaptic plasticity and subcellular localization of NADPH-diaphorase (NADPH-d), a cytochemical marker for nitric oxide syntase, in the pedal ganglia of the Gray mussel Crenomytilus grayanus sampled from the polluted and reference sites in Amursky Bay (Sea of Japan) at lower and higher water temperature (in the beginning and the end of August, respectively). At lower temperature, neuroplastic changes in mussel ganglia prevailed: a sharp increase in the number of cytosomes in NADPH-d-positive neurons and a sharp decrease in the number of mitochondria in both NADPH-d-positive and NADPH-d-negative neurons. At higher temperature, neurodegenerative changes prevailed: disruption of a part of NADPH-d-negative axons and interneuronal contacts, formation of concentric lamellar structures in the neuropils, and accumulation of autophagosomes in NADPH-d negative neurons. The results suggest that the stress-induced production of nitric oxide in cytosomes of mussel neurons and plasticity of gap junctions have a neuroprotective effect.     

Localization of 17 beta-hydroxysteroid dehydrogenase in the gonads of bivalve mollusks--the sea pecten (Patinopecten yessoensis Jay) and Gray's mussel (Crenomytilus grayanus Dunker)

Presence of 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) in testes and ovaries of the common mussels--Patinopecten yessoensis (Jay) and Crenomytilus grayanus (Dunker) has been demonstrated histochemically. The enzyme is revealed in some granular amebocytes and germ cells. In growing oocytes its activity is higher that in oocytes completed their growth. 17 beta-HSD is revealed electron microscopically near agranular endoplasmic reticulum, or on the external surface of its membranes and in globules, possessing, evidently, lipid nature. The data obtained demonstrate that synthesis and metabolism of steroid hormones are possible both in additional gonadal elements and in germ cells of the animals investigated.     

The Effect of Metal Concentration in Bottom Sediments on the Accumulation of Metals by the Mytilids Crenomytilus grayanus and Modiolus kurilensis

The relationship between the concentration of Pb, Cu, Cd, Zn, and Ag in soft tissues of mytilids Crenomytilus grayanus and Modiolus kurilensis and in bottom deposits was studied to determine the potential and limitations in the use of these molluscs as bioindicators of contamination of coastal waters near Vladivostok (northwest of the Sea of Japan). It was found that the mussel C. grayanus began to accumulate Pb and Cu after certain threshold concentrations of these metals (60 and 150 g/g respectively) in fine fractions of sediments was reached. Accumulation of Pb and Cu by M. kurilensis also depended on the level of sediment contamination, but the higher initial metal content in tissues and the higher individual variability reduced the sensitivity of this species as a bioindicator. Accumulation of Cd and especially of Zn in mytilid tissues is adjusted by the organism, which restricts the opportunity of their use for tracking the anthropogenic input of bioavailable forms of these metals.     

Sperm Ultrastructure of Two Mollusk Species Geukensia demissa and Modiolus modiolus (Mytilidae, Bivalvia)

Sperm ultrastructure was studied in two Atlantic species of bivalve mollusks Geukensia demissa and Modiolus modiolus (Mytilidae). These spermatozoa have a classic structure common for species with external fertilization: the head of the spermatozoon of G. demissa and M. modiolus consists of an acrosome, including an acrosomal vesicle and periacrosomal material, a nucleus, and a middle part with two mutually perpendicular centrioles, surrounded by a ring of spherical mitochondria. The spermatozoa of G. demissa and M. modiolus differ in the general shape and size of the head (M. modiolus, 5.8 × 4 m; G. demissa, 4.2 × 2.5 m), as well as the structure of the acrosome and the number of mitochondria (M. modiolus, 8–12; G. demissa, 5–6). The spermatozoa of the Atlantic species M. modiolus and M. modiolus (M. kurilensis) from the Sea of Japan are close in ultrastructure, but differ in the number of mitochondria. This could possibly be a reason to consider M. modiolus from the Sea of Japan as a separate subspecies, M. modiolus kurilensis. In ultrastructure, the spermatozoa of G. demissa differ significantly from the spermatozoa of M. modiolus, but are similar to the spermatozoa of species of the genus Brachidontes.     

Mitotic chromosomal abnormalities and DNA polymorphism in the Nile tilapia (Oreochromis niloticus,Linnaeus, 1758) as a biomarker for water pollution by heavy metals

Mitotic chromosomal aberrations and DNA polymorphism (RAPD marker) were carried out on the Nile tilapia Oreochromis niloticus collected from five sites in Minia governorate, Egypt to test their applicability as biomonitors for heavy metal contaminants of water. The diploid chromosome number of O. niloticus population was 2 n = 44. Different types of chromosomal aberrations were recorded (e.g., deletion, ring, centromeric attenuation, end-to-end association, dicentric chromosome, stickiness chromosomes, endomitosis, fragments and chromatid gap). The chromosomal aberrations varied between O. niloticus population collected from five sites, and the most common type was ring (R) chromosomes. Samples obtained from Bahr Yousef and Irrigation drain exhibited the highest aberration frequency. The frequency of chromosomal aberration was positively correlated with the concentration of heavy metals where their concentration in the surface water of Irrigation drain and Bahr Yousef exceeded the limits defined by WHO as well as the concentration of Pb in muscles. The RAPD marker was also used to identify genetic variation among Nile tilapia samples collected from five different water sources. It created polymorphic and unique bands that can be used as genetic markers to track DNA variations. The dendrogram also revealed that exposure to heavy metal pollution causes gradual accumulation of variance, whereas areas subjected to environmental stress showed higher genetic variation and clustered together.     

A comparison of relative growth in Cerastoderma (=Cardium) edule, Modiolus modiolus, and Mytilus edulis (Mollusca: Bivalvia)

Relative growth has been studied in intertidal populations of Cerastoderma edule and Mytilus edulis and in a subtidal population of Modiolus modiolus. Size dimensions were generally more variable and shell thickness generally greater in the intertidal species. Total weight was correlated with shell weight in all three species but there was a progressively greater emphasis on tissue growth through the sequence Cerastoderma, Mytilus, Modiolus. These results are interpreted in terms of differences in stability between their respective habitats.
Analyses of individual year classes in Cerastoderma showed a marked pattern of alternating years of good and poor growth.     

Koi herpesvirus infection in experimentally infected common carp Cyprinus carpio (Linnaeus, 1758) and three potential carrier fish species Carassius carassius (Linnaeus, 1758); Rutilus rutilus (Linnaeus, 1758); and Tinca tinca (Linnaeus, 1758) by quantitative real‐time PCR and in‐situ hybridization

Only single cells in the carrier fish species Carassius carassius (Linnaeus, 1758) for koi herpesvirus (KHV) are infected in contrast to large numbers in the susceptible species common carp Cyprinus carpio (Linnaeus 1758). Several species of the family Cyprinidae have been described as virus carrier species, showing no clinical signs of a KHV disease but able to transmit the virus to other susceptible fish. In this study, 72 common carp Cyprinus carpio (Linnaeus, 1758), 36 tench Tinca tinca (Linnaeus, 1758), 36 crucian carp Carassius carassius (Linnaeus, 1758) and 36 common roach Rutilus rutilus (Linnaeus, 1758) were experimentally infected with KHV (isolate “Israel”) by immersion and kept at 20°C. The fish were euthanized at 12 timepoints over a period of 90 days and virus DNA was quantified in tissues by a real‐time TaqMan PCR. Whereas KHV‐DNA was found in Cyprinus carpio for up to 90 days, the virus DNA was detectable only in single individuals of Rutilus rutilus, Tinca tinca and Carassius carassius for up to 25 days after experimental virus exposure. Tissue samples of Cyprinus carpio and Carassius carassius were screened by in‐situ hybridization. Positive signals were found in various organs of the common carp tested crucian carp. In the latter species a much smaller number of virus‐positive stained cells was detected compared to the infected carp.     

Localization of NO-Ergic Structures in Oral Lobes, Labia, and Esophagus of the Bivalve Mollusc Crenomytilus grayanus

Localization of NO-ergic elements in oral lobes, labia, and esophagus in the bivalve mollusc, mussel Crenomytilus grayanus was studied using histochemical technique [1] for detection of NADPH-diaphorase (EC 1.6.99.1). The NO-producing elements were revealed in all studied parts of the digestive system. NADPH-diaphorase was found in nerve and secretory cells as well as in nerve plexuses. Numerous NO-ergic nerve cells were observed in the basal part of epithelium of labia and of the initial part of esophagus as well as in the subepithelial area of these organs. In the middle and posterior parts of esophagus, only subepithelially located NO-ergic nerve cell are present. Basiepithelial NO-producing secretory cells are found in all the parts, but most of these cells are observed in labia and the initial part of esophagus. Subepithelial secretory cells labeled with diformazan granules are spread from the folded surface of oral lobes to the initial part of esophagus; no such cells were found on the smooth surface of the lobes. The deposit labeled basi- and subepithelial nerve plexuses in all studied organs except for oral lobes. These plexuses are the most developed in labia and the initial part of esophagus of the studied mollusc.     

Reproduction efficiency of the Horse Leech,Haemopis sanguisuga (Linnaeus, 1758)

The aquaculture sector has recently focused on alternative food sources due to increased costs and limited resources. Live food sources, such as invertebrates, have important roles in digestion processes of fish. It was aimed to determine the reproduction efficiency of the horse leech, Haemopis sanguisuga, which is expected to be used as an alternative live nutritional source in aquaculture in this study. The study was conducted at ambient temperature in the laboratory conditions for three months. A total of 25 gravid leeches with an average body mass of 6.91 ± 2.20 g were used. The leeches lost up to 75% of their body mass and laid 4.30 ± 1.49 cocoons at 12.0 ± 8.2 days interval during cocoon laying period. Leeches stopped laying cocoon when the temperature fell to 16 °C. The average of offspring hatched from the cocoons was 6.45 ± 2.86. The length–body mass relationship of the offspring was exponential (W = 0.012L^2.809, R² = 0.898). The results from the current study indicate that the basic biological parameter affecting the reproduction efficiency of H. sanguisuga is the size of the broodstock leeches, and reproduction activity weakens the broodstocks.