Invertebrate Survival Journal

Involvement of GSK-3β in regulating endoplasmic reticulum stress-mediated apoptosis in Patinopecten yessoensis under high temperature stress

K Zhou, Z Tong, C Yang, Z He, L Wang, L Song — Mon, 11 Aug 2025 00:00:00 +0000

Glycogen synthase kinase-3β (GSK3β), a conserved regulator involved in glycogen metabolism and signal transduction, was identified in Yesso scallop (Patinopecten yessoensis) as PyGSK-3β, containing a conserved S_TKc domain and phosphorylation sites at Ser9 and Tyr216. The gene expression, protein phosphorylation and apoptosis under high temperature (HT) stress were assessed by using quantitative real-time PCR, Western blotting, and immunohistochemistry. PyGSK-3β mRNA was ubiquitously expressed in all the examined tissues and haemocytes. HT stress significantly upregulated the mRNA expression of PyGSK-3β, PyGRP78, PyIRE1, PyJNK, and PyCaspase-3, as well as the phosphorylation level of PyGSK-3β (Tyr216) and PyJNK (Thr183) in gills, and decreased the mRNA expression of PyBCL-2. Furthermore, the GSK-3 inhibitor SB216763 significantly suppressed the mRNA expression of PyGSK-3β, PyGRP78, PyJNK and PyCaspase-3, decreased the phosphorylation level of PyGSK-3β (Tyr216) and PyJNK (Thr183), and reduced HT stress-induced cell apoptosis after HT treatment. However, SB216763 had no effect on the mRNA expression of PyBCL-2. These findings highlight the pivotal role of PyGSK-3β in ER stress-mediated apoptosis and offer insights into scallop high temperature stress adaptation, suggesting its potential as a molecular target for breeding or further functional studies in aquaculture.

Effects of Magnetic Iron Oxide Nanoparticles (Fe3O4 NPs) on Galleria mellonella Hemocytes

AN Eskin, Ş Öztürk, A Eskin — Thu, 31 Jul 2025 00:00:00 +0000

In this study, we examined the harmful effects of magnetic iron oxide nanoparticles (Fe₃O₄ NPs) on the Greater Wax Moth Galleria mellonella (L.) (Lepidoptera: Pyralidae). For this aim, G. mellonella larvae were administered Fe₃O₄nanoparticles (NPs) at concentrations of 0.4, 2, 10, 50, and 250 µg/10 µl. Subsequently, the impact of these concentrations on total hemocyte counts (THCs) and various hemocyte indices, namely viable, mitotic, apoptotic, necrotic, and micronucleated cells, was assessed using hematoxylin and eosin (H&E) staining. Compared to the control group, G. mellonella larvae exposed to Fe₃O₄ NPs at concentrations of 10, 50, and 250 µg/10 µl exhibited a statistically significant reduction in THCs. Additionally, in these experimental groups exposed to different concentrations of Fe₃O₄ NPs, the percentage of viable cells significantly decreased, while the percentage of apoptotic cells increased in comparison to the control group. Likewise, the percentage of necrotic hemocytes was significantly higher in the larvae exposed to 50 and 250 µg/10 µl Fe₃O₄ NPs than that in the control group. Furthermore, the frequency of micronucleated hemocytes was significantly elevated at Fe₃O₄ NP doses of 10 and 50 µg/10 µl when compared to the control group.

Transcriptome analysis of Apostichopus japonicus early in the regeneration process after evisceration

BC Wu, HF Dang, XH Wang, Q Li, W Zhou, J Liu, SG Ye, RJ Li — Thu, 19 Jun 2025 00:00:00 +0000

In extreme salt stress and anaerobic conditions, Apostichopus japonicus survives by "evisceration." The species can eject its viscera and create a new organ if the habitat is suitable. The molecular response of A. japonicus coelomocytes and visceral regeneration is examined. Previous research found A. japonicus coelomocyte volume recovery 2 h-p-e (two hours post-evisceration) mirrored fast human hematopoiesis. Coelomocytes returned to pre-evisceration levels 6 h-p-e (six hours post-evisceration). First, KCL solution was artificially injected at A. japonicus body mass. Subsequently, evisceration occurred. Immunological tissue coelomocytes were taken at 2 h and 6 h-p-e, and those from the control group (0 h) were also collected. Coelomocyte transcriptome was sequenced. Transcriptome sequencing was applied to the collected coelomocytes. At 2 h-p-e, 860 differentially expressed genes were identified, with 639 upregulated and 221 downregulated. At 6 h-p-e, 1638 genes were upregulated and 432 downregulated. Immunity genes were predominantly differentially expressed in the ‘Molecular function’ category, the major category for these immunity - related differentially expressed genes. Gene differences were analyzed using KEGG annotation. At 2 h-p-e, by comparing the gene expression of A. japonicus coelomocytes with that of the non - eviscerated control group (0h), 860 DEGs were identified. These DEGs were associated with 53 pathways, 22 of which were related to illnesses and the immune system, like the ECM - receptor interaction pathway. 6 h-p-e, 20 major enrichment routes were selected, and 11 of them, including the Phagosome pathway, were associated with regeneration. [Abstract cut for lenght]

Toxicity of the two insect growth regulators, pyriproxyfen and Lufox® against cotton bollworm

M Khorshidi, R Farshbaf Pour Abad, A Zibaee, M Saber — Tue, 25 Feb 2025 00:00:00 +0000

This study investigated the field effects of two insect growth regulators, pyriproxyfen and lufenuron+fenoxycarb (A commercial combination), on some biological and physiological characteristics of third instar larvae of the cotton bollworm, Helicoverpa armigera Hubner. The insecticides significantly affected the weight and duration of larval and pupal development, the longevity of males and females, the egg laying and hatching rates. The larvae treated with sublethal concentrations showed an increase in the total number of hemocytes, plasmatocytes and granulocytes. A significant increase in aminotransferase activity was also observed in the hemolymph of treated larvae. The increased activity of acid phosphatase and alkaline phosphatase was also observed in the treated H. armigera larvae. The total activity of esterase and glutathione S-transferase as detoxifying enzymes also increased significantly after pyriproxyfen and lufenuron+fenoxycarb treatments compared to control. In contrast, the amount of storage macromolecules, protein, triglycerides and glycogen, which are related to the energy cost of pesticide treatment, decreased in the treated larvae compared to the control. According to the results of the present research, the pesticides pyriproxyfen and lufox can be used to control the population of H. armigera by interfering with the intermediate metabolism and cellular immunity of the larvae, but their efficiency and compatibility should be considered with other control methods, especially biological factors.

The effect of Artemisia annua essential oil and one of its main components on the biology and enzymatic and non-enzymatic activities of Ephestia kuehniella Zeller (Lepidoptra: Pyralidae)

S Beshkoufe, R Azizi, Z Afrazeh, A Nemati, M Mojarab-Mahboubkar, J Jalali Sendi — Thu, 31 Jul 2025 00:00:00 +0000

The flour moth, Ephestia kuehniella, is a common storage pest worldwide. One of the most commonly used methods for controlling storage pests is fumigation using synthetic chemical compounds, which have serious negative impacts. In this study, the fumigant toxicity effects of Artemisia annua essential oil and 1,8-cineole, two environmentally safe substances, were investigated on larvicidal activity, developmental stages, cellular immunity, and enzymatic activities of E. kuehniella larvae. The results of fumigation assays on 3^rd instar larvae after 24 and 48h was estimated for A. annua essential oil and 1,8-cineole and lethal concentration (LC) values was calculated. The results indicated higher toxicity of 1,8-cineole over A. annua essential oil. Moreover, prolongation of larval developmental duration and a decrease in pupation period were observed at LC₃₀-treated larvae with A. annua essential oil as well as 1,8-cineole. A significant reduction of total hemocyte count and differential hemocyte count, including the number of plasmatocytes and granulocytes was also observed at LC₃₀ and LC₅₀ concentrations, 24 and 48h post-treatment. The antioxidant enzymes including catalase and peroxidase were significantly increased compared to the control. Similarly, the metabolic enzymes including alanine aminotransferase, aspartate aminotransferase, acid/alkaline phosphatase, lactate dehydrogenase were enhanced. The evaluation of detoxifying enzymes like glutathione S-transferase using DCNB and CDNB substrates, showed a significant increase compared to the control. Conversely, the activity of acetylcholinesterase was significantly decreased compared to the control. The results are indicative of the potentiality of these natural compounds as alternative to classical control measure against this important storage product pest

Influence of sex on intestinal microbiota of Pacific white shrimp Penaeus vannamei

G-Z Wang, Y-X Wang, M-Q Wang — Mon, 21 Jul 2025 00:00:00 +0000

Penaeus vannamei is a high-value aquaculture species. However, with the expansion in farming area and the rise in stocking density, the disease problems of P. vannamei have grown increasingly severe. Given the pivotal role of intestinal microbiota in regulating host health, including digestion, immune function, and metabolic homeostasis, understanding microbial dynamics is critical for disease control. Notably, although pronounced sexual dimorphism exists in P. vannamei, gender-specific microbiota variations remain uncharacterized. This study intends to evaluate the impact of gender variations on the intestinal microbiota of P. vannamei, using the 2bRAD-M technique. The results showed that the average growth rate of female shrimp was significantly higher than that of male shrimp. The Chao 1 index and Simpson index of female shrimp were greater than those of male shrimp. β-diversity research suggested that the female group samples might contain more microbial variety. At the phylum level, the microbial composition of the female and male shrimp groups is similar, with Pseudomonadota, Bacillota_A, Bacteroidota, Actinomycetota, and Planctomycetota being the dominating phyla. At the species level, the female group is predominantly formed of Phaeobacter italicus, NSJ_50_sp014385105, Pseudoalteromonas spongiae, and Xanthomarina gelatinilytica, while the male group contains a larger abundance of Vibrio parahaemolyticus and Vibrio cholerae. These data indicated that female P. vannamei not only display faster growth rates, but also possess a more complex and diverse intestinal microbiota, which may contribute to their higher disease resistance compared to male ones.

XXV scientific meeting of the Italian Association of Developmental and Comparative Immunology (IADCI), February 19-21, 2025, Department of Sciences and Technological Innovation - DISIT, University of Eastern Piedmont “Amedeo Avogadro”, Alessandria, Italy

F Dondero, A Calisi — Thu, 19 Jun 2025 00:00:00 +0000