Search Results (26)

Cisplatin is a highly cytotoxic drug used for the treatment of head, neck, and soft tissue cancers; however, it has nephrotoxic effects that can lead to acute kidney injury. Protocatechuic acid (PCA) is a natural widely available antioxidant found in many fruits such as kiwi, mango, and berries. We have recently shown that PCA reduced renal injury in a mouse model of unilateral ureteral obstruction. The current study aims to investigate the protective effects of PCA in Cisplatin-induced inflammation in vitro in Boston University Mouse Proximal Tubular (BUMPT) cells. BUMPT cells were cultured in complete DMEM. Confluent BUMPT cells were then treated with 20 μM Cisplatin ± PCA 50 or 100 μM for 24 h. PCA treatment showed a dose-depending increase in % cell viability in Cisplatin-treated BUMPT cells. PCA treatment also dose-dependently decreased Cisplatin-induced increases in oxidative stress (ROS and TBARS), inflammation (p-NF-κB and IL-6), and apoptosis (cleaved caspase-3 and % of TUNEL⁺ cells) compared to Cisplatin-only treatment. The reduction in oxidative stress, inflammation, and apoptosis with PCA treatment in Cisplatin-treated BUMPT cells was associated with decreases in tubular physical barrier resistance and the expression of the tight junction protein zonula occludens-1 (ZO-1) when compared to BUMPT cells treated with Cisplatin alone. The current findings suggest that PCA treatment improves tubular barrier function in Cisplatin-treated BUMPT cells via reductions in oxidative stress, inflammation, and apoptosis. Full article

Background: Snapper is a significant commercial, recreational, and cultural teleost species in New Zealand, with aquaculture potential. The impact of long-term (chronic) temperature changes on immune and stress responses have not been studied in snapper, yet they have a critical importance to the health status of the fish. Methods: We investigated a set of genes in 30 individual snapper including fin, head kidney, and liver tissue, fish (10 per group) were exposed to either warm (22 °C), cold (14 °C), or ambient temperatures (10.5–18.6 °C) for 3 months. Results: Analyses of experimental fish using NanoString technologies to assess stress- and immune-related genes in the three tissue types showed that 22 out of 25 genes changed significantly in the experiment, indicating the significant impacts of chronic temperature changes on stress and immune responses. Furthermore, using a combined dataset based on this study and a previous one testing the impact of acute temperature changes in snapper, we identified five genes in the non-lethal fin-clip samples that can predict internal organ health status. Conclusions: Taken together, our experiments demonstrate the potential of the NanoString gene expression assessment tool for the rapid monitoring of stress responses in snapper, which can aid in the selection of stress-resilient wild stocks, monitor species in aquaculture environments, and inform the selection of locations for aquaculture. Full article

To explore the effect of density on large yellow croaker (Larimichthys crocea) under intensive aquaculture conditions and determine the appropriate culturing density, this study investigate the effects of different stocking densities on the nutritional composition, stress, and immune levels of large yellow croaker. Through a long-term aquaculture experiment, conducted under flow-through conditions of intensive aquaculture, three initial density groups were set: a low density group [LD], 4.92 kg/m³; a medium density group [MD], 7.56 kg/m³; and a high density group [HD], 10.08 kg/m³, for a 150-day rearing trial. Large yellow croaker were fed to satiation twice daily (6:00, 17:00). At the end of this trial, the final densities were 10.38 ± 0.50, 14.41 ± 1.06, and 18.71 ± 0.99 kg/m³ in the LD, MD, and HD groups, respectively. The results showed that the growth performances were adversely influenced by a high stocking density. Levels of cortisol in serum, superoxide dismutase (SOD) and catalase (CAT) in liver, Na⁺-K⁺ ATPase and Na⁺-K⁺ ATPase gene in gills, and heat shock protein (HSP70/90) genes and glutathione S-transferase (GST) genes in the liver significantly increased under HD treatment. Results of immune response analyses showed that there was a clear decrease in immunoglobulin M (IgM), complement component 4 (C4), and lysozyme (LZM) in serum, lysozyme (LZM) genes, tumor necrosis factor-alpha (TNF-α) genes and interleukin-1β (IL-1β) genes in the head kidney of large yellow croakers reared in the HD group. An obvious increase in free amino acids and fatty acids in the muscle of large yellow croakers reared in HD group was also observed. Overall, this study showed that the optimal final culturing density of large yellow croaker under flow-through systems should be between 14.41 kg/m³ and 18.71 kg/m³ to improve aquaculture efficiency and product quality. Full article

Aeromonas hydrophila causes motile Aeromonas septicemia (MAS), a disease with a high mortality rate in tilapia culture. Feed-based vaccines with the incorporation of inactivated whole-cell bacteria into the feed offer promising tools to control MAS. Currently, the incorporation of genome-free bacteria as bacterial vaccine through the implementation of SimCells^® technology into the feed has become a particular interest. Background/Objectives: This study investigates the efficacy of a feed-based vaccine incorporating genome-free A. hydrophila (FBV-GFAH) against MAS infection in red tilapia. Methods: The vaccine was prepared and delivered at 5% fish body weight for three consecutive days in weeks 0 (prime vaccination) and 2 (first booster vaccination), orally. Throughout a five-week experimental period, the immune-related genes (IL-1β, MHC-II, CD4, IgT, and IgM) expression in the hindgut and head kidney of the fish was determined using RT-qPCR assay. Lysozyme (serum) and overall IgM (serum, gut lavage, and skin mucus) productions were also detected. Results: Fish vaccinated with FBV-GFAH showed a significant (p ≤ 0.05) improvement in relative percent survival compared with unvaccinated fish following bacterial challenge. FBV-GFAH induced the expression of immune-related genes in the hindgut and head kidney, especially after booster vaccination. Furthermore, serum lysozyme activity and overall IgM production in serum, skin mucus, and gut lavage were also significantly (p ≤ 0.05) improved in the FBV-GFAH vaccinated fish than the unvaccinated fish. Conclusions: This study showed that FBV-GFAH is a promising feed-based vaccine technology to control MAS in cultured tilapia. Full article

Background: Snapper (Chrysophrys auratus) is a commercially, recreationally and culturally important teleost species in New Zealand and has been selected as a potential new species for aquaculture. Selective breeding to enhance stress tolerance, survival and growth are major breeding targets, yet research into snapper immune and stress responses has been limited. Methods: We explored a set of candidate genes in the fin, head kidney and liver tissues of 50 individuals by exposing 20 fish to increasing temperature (up to 31 °C) and 20 fish to decreasing temperature (down to 7 °C) for up to 37 h. Of these, we analysed 10 temperature-sensitive and 10 temperature-tolerant fish, along with 10 fish kept at 18 °C (acclimation temperature) as a control group. Results: Expression analyses of candidate stress genes in the three tissue types via NanoString Technologies, Inc., Seattle, WA, USA. showed that 20 out of 25 genes significantly changed in each experiment, demonstrating the significant impact of temperature on stress and immune responses. We further document that 10 key gene biomarkers can be used to predict genotypes that are tolerant to extreme temperatures. Conclusions: Taken together, our novel NanoString method can be used to monitor stress in snapper rapidly, and applications of this tool in this and potentially closely related teleost species can provide insights into stress resilience of wild stocks and inform the selection of grow-out locations for aquaculture. Full article

Phosphoinositide 3-kinases (PI3Ks) are a class of key regulatory factors in eukaryotes that can inhibit viral replication by influencing autophagy. Currently, cyprinid herpesvirus 3 (CyHV-3) poses a serious threat to common carp culture. However, PI3K has not yet been identified in common carp. In this study, full-length PI3KC3 from common carp (CcPI3KC3), consisting of an open reading frame (ORF) of 2664 bp encoding a polypeptide of 887 amino acids, with a predicted molecular mass of 101.19 kDa and a theoretical isoelectric point (pI) of 5.97, was cloned. The amino acid and nucleotide sequences of CcPI3KC3 displayed high similarity to yellow catfish’s (Tachysurus fulvidraco) PI3KC3. The tissue expression profile revealed that the mRNA levels of CcPI3KC3 in the liver, spleen, and head kidney were significantly greater than those in the brain, heart, intestines, gills, eyes, testes, and ovaries of common carp. We compared the expression patterns of CcPI3KC3 between “Longke-11” mirror carp (CyHV-3-resistant carp) and German mirror carp (non-resistant to CyHV-3) at different times (0, 48, 96, 144 h, 192, 240, 288 h post-infection (hpi)) after CyHV-3 infection. The results revealed that CcPI3KC3 mRNA expression significantly increased in the early infection stage. In the CyHV-3-resistant mirror carp variety, the relative expression of CcPI3KC3 was significantly greater at 48, 96, and 144 hpi compared with the nonbreeding strain groups after infection (p < 0.001). These results indicate that the full-length CcPI3KC3 sequence was successfully cloned from common carp for the first time, and it might play an important role in the immune system of common carp against CyHV-3 infection. This study provides a theoretical basis for the molecular mechanism of CyHV-3 resistance. Full article

Several exogenous probiotics are applicable in fish culture; however, challenges in isolation and verification have hindered the full utilization of numerous host probiotics. Therefore, this study aimed to apply the host probiotic Exiguobacterium acetylicum G1-33 to hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) cultures and explore its mechanism of action. In total, 360 hybrid grouper were divided into four groups, which were fed the following for 60 days: three received commercial feed with varying concentrations of E. acetylicum G1-33 (10⁶, 10⁸, and 10¹⁰ CFU/g), while a control group received commercial feed. The results showed that supplementation with 10⁶ and 10⁸ CFU/g of E. acetylicum G1-33 enhanced gut morphology, upregulated growth-related genes (ghr1, igf-2, s6k1, tor), and promoted growth, with supplementation with 10⁸ CFU/g resulting in the most notable enhancement. However, supplementation with 10¹⁰ CFU/g inhibited growth, possibly because of changes in intestinal morphology. Additionally, supplementation with E. acetylicum G1-33 upregulated the expression of immune-related genes (c3, myd88, Cu/Zn-sod, tlr3, and tnf2) in the liver and head kidney but led to an increase in malondialdehyde content, as well as a decrease in alkaline phosphatase and acid phosphatase activities, in the liver and serum, indicating increased oxidative stress. Moreover, supplementation with 10⁶ and 10⁸ CFU/g E. acetylicum G1-33 enhanced the widespread expression of immune-related genes in the head kidney and liver, respectively, and improved resistance to Vibrio harveyi, whereas supplementation with 10¹⁰ CFU/g weakened this resistance. In conclusion, E. acetylicum G1-33, particularly at 10⁸ CFU/g, emerged as an effective probiotic, optimizing growth performance and immunity in hybrid grouper. This research is pioneering in its application of E. acetylicum in mariculture, potentially broadening the range of probiotic strategies in aquaculture. Full article

As an important research tool, cell lines play a vital role in life science research, medical research, and drug development. During the culture of the Scophthalmus maximus head kidney (TK) cell line, we found a phenomenon of cell vacuolization caused by excessive serum concentration. Moreover, the vacuolization of the cells gradually disappeared after passage by trypsin digestion. In clarifying the formation mechanism of this reversible cellular vacuolation, transcriptomics was utilized to explore the mechanism of cell vacuolization caused by excessive serum concentration. Transcriptome analysis indicated that excessive serum concentration could cause the up-regulated expression of PORCN and other genes to promote cell proliferation. Compared with cells whose vacuolization disappeared after trypsin digestion and passage, the expression of mitosis-related genes (BUB1, ttk, Mad2, Cdc20, CDK1, CCNB1), nuclear stability-related genes LMNB1 and tissue stress and repair-related genes HMMR in vacuolated cells caused by excessive serum concentration was significantly up-regulated. There is a regulatory system related to adaptation and stress repair in the cells, which can maintain cell stability to a certain extent. This study provides a theoretical basis for the stable culture of fish cell lines and the solution to the problem of cell vacuolation. Full article

Green terror cichlid (Aequidens rivulatus) is a popular tropical freshwater ornamental fish. In 2021, an unknown disease was observed in cultured A. rivulatus in Tianjin, China, with a cumulative mortality rate of 25% within 7 days of onset. The main clinical signs were scale loss, skin ulceration, and slight bleeding. Histopathological observation revealed obvious damage to the liver, spleen, and kidney of diseased fish. In addition, abundant granulomas were observed in the spleen and head kidney of the diseased fish. To define the potential pathogens from A. rivulatus, bacteria were isolated from the visceral tissue of diseased fish with conventional methods. An artificial infection experiment was carried out to prove the pathogenicity of the isolated bacteria. The strains HG-2021-1 and HG-2021-3 were isolated from diseased fish and identified as being responsible for the disease. They were identified as Streptococcus iniae based on physiological and biochemical tests, lctO gene detection, and 16S rRNA gene sequence analysis. According to the result of multilocus sequence typing (MLST), HG-2021-1 and HG-2021-3 belong to different genotypes of S. iniae. Furthermore, they were found to contain the virulence genes pgmA, scpI, cpsD, and pdi, and the median lethal dose (LD₅₀) for A. rivulatus was 1.8 × 10⁶ Colony-Forming Units (CFU)/mL and 6.6 × 10⁶ CFU/mL, respectively. To our knowledge, this is the first report of fish coinfected by two genotypes of S. iniae. Full article

Aquaculture fish are kept for long periods in sea cages or tanks. Consequently, accumulated stress causes the fish to present serious problems with critical economic losses. Fish food has been supplemented to reduce this stress, using many components as amino acids such as tryptophan. This study aims to determine the transcriptional effect of tryptophan and cortisol on primary cell cultures of salmon head and posterior kidney. Our results indicate activation of the kynurenine pathway and serotonin activity when stimulated with tryptophan and cortisol. An amount of 95% of tryptophan is degraded by the kynurenine pathway, indicating the relevance of knowing how this pathway is activated and if stress levels associated with fish culture trigger its activation. Additionally, it is essential to know the consequence of increasing kynurenic acid “KYNA” levels in the short and long term, and even during the fish ontogeny. Full article

Fish cell culture is a common in vitro tool for studies in different fields such as virology, toxicology, pathology and immunology of fish. Fish cell cultures are a promising help to study how to diagnose and control relevant viral and intracellular bacterial infections in aquaculture. They can also be used for developing vaccines and immunostimulants, especially with the ethical demand aiming to reduce and replace the number of fish used in research. This study aimed to isolate head kidney primary cell cultures from three Chilean salmonids: Salmo salar, Oncorhynchus kisutch, and Oncorhynchus mykiss, and characterize the response to bacterial and viral stimuli by evaluating various markers of the innate and adaptive immune response. Specifically, the primary cell cultures of the head kidney from the three salmonids studied were cultured and exposed to two substances that mimic molecular patterns of different pathogens, i.e., Lipopolysaccharide (LPS) (bacterial) and Polyinosinic: polycytidylic acid (POLY I:C). Subsequently, we determined the mRNA expression profiles of the TLR-1, TLR-8, IgM, TLR-5, and MHC II genes. Head kidney primary cell cultures from the three species grown in vitro responded differently to POLY I:C and LPS. This is the first study to demonstrate and characterize the expression of immune genes in head kidney primary cell culture isolated from three salmonid species. It also indicates their potential role in developing immune responses as defense response agents and targets of immunoregulatory factors. Full article

Golden pompano, Trachinotus ovatus, is a perciform fish with great economic value and is widely cultured in the coastal areas of China. The wide occurrence of bacterial, parasitic and viral diseases has seriously threatened the sustainable development of the golden pompano culture industry. Janus kinases (JAKs) play important roles in most cytokine-mediated inflammatory responses, antiviral immune responses, autoimmune responses and disease pathologies. The present study aimed to obtain the full-length cDNA sequences of JAKs (JAK1, JAK2a, JAK3 and TYK2) from golden pompano and investigate their roles following stimulation with lipopolysaccharide (LPS), polyriboinosinic-polyribocytidylic acid (poly I:C) and Vibrio alginolyticus using RT-PCR, RACE-PCR and real-time qPCR methods. All four JAK proteins of golden pompano shared similar conserved domains, had high identities and clustered well with their teleost counterparts in phylogenetic analysis. Furthermore, these four genes were expressed in all examined tissues from healthy fish and induced in head kidney (HK), spleen, liver and gill post LPS, poly I:C and V. alginolyticus stimulation. Knowledge of the roles of JAKs in the immune response to different microbial pathogens provides a basis for further understanding of these functions. Full article

Cladosporium infections have a poor prognosis in animals, most likely due to a lack of knowledge about diagnosis and treatment. In this study, we described a case of a lethal Cladosporium allicinum infection in a captive bullfrog (Pyxicephalus adspersus) in Europe. One adult male bullfrog was referred with clinical signs of lethargy and a cutaneous nodule. Fungal infection was suspected on cytology and confirmed by histology and cultural isolation. The mold was identified by molecular methods using partial sequencing of the TEF1α gene and the ITS region of rDNA. Climbazole antifungal treatment was started but the frog died after 30 days, and necropsy was done. Pigmented hyphae and structures consistent with muriform bodies were found on a background of diffuse granulomatous inflammation at cytological and histopathological examinations. Fungal culture revealed the presence of pigmented fungi identified as Cladosporium allicinum only by partial sequencing of the TEF1α gene. A focally extensive granuloma with intralesional hyphae and muriform bodies effacing the architecture of head, liver, kidneys, lungs, and large intestine were retrieved after necropsy. This study is the first Italian report of the occurrence of lethal C. allicinum infection in a frog and highlights the role of this Cladosporium sp. in chromoblastomycosis. Full article

The golden pompano (Trachinotus ovatus) is one of the most economically valuable marine fishes in South China. Streptococcus agalactiae, an infectious Gram-positive bacterium that is highly destructive for golden pompano culture, has recently caused massive losses to the golden pompano industry. This study aimed to investigate the dynamic immune response of golden pompano to S. agalactiae infection, using RNA-seq analysis at two different time points after infection. Abundances of differentially expressed genes (DEGs) gradually increased in the liver and spleen 48–120 h post-infection, whereas those in the head kidney were lower at 120 h than at 48 h. Pathway enrichment analysis of DEGs revealed that genes related to the complement system were continuously transcribed between 48 and 120 h. Metabolic and immune-regulation-related pathways were highly enriched in the liver 48 h after infection. Transcriptome analysis was verified using quantitative PCR for eight genes with similar expression trends. This study revealed the inflammatory response of golden pompano after S. agalactiae infection, including inflammation-related chemokines and signaling pathways. Our findings provide a theoretical basis for studying S. agalactiae resistance in golden pompano and provide a reliable resource for the genetic breeding of fish. Full article

In this study, a novel interleukin-1-family member (nIL-1Fm) was studied from the Nile tilapia (Oreochromis niloticus), a widely cultured fish species in brackish water and freshwater in Thailand and worldwide. The complete sequence of the Nile tilapia nIL-1Fm comprised 2474 bp, encoding 350 amino acids. The genome organization of nIL-1Fm had seven exons and six introns, which were conserved in teleost fish. During tissue distribution analysis, the nIL-1Fm was extremely increased in the head kidney, tailfin and trunk kidney, which had the highest levels, followed by the spleen. Additionally, nIL-1Fm was significantly up-regulated under PAMP stimulation, whereas the stimulation of LPS, PHA and poly I:C could significantly induce nIL-1Fm expression in vitro. Moreover, nIL-1Fm transcription was significantly induced when infected with Streptococcus agalactiae in the spleen, liver and intestine. Taking together, our results indicated that nIL-1Fm can expand our knowledge of the IL-1 cytokine family in fish immune response. Full article