Anterior Kidney Research Articles

Bioaccumulation of arsenic in fish (Labeo rohita) in presence of periphyton: ameliorative effect on oxidative stress, physiological condition, immune response and risk assessment.

The present study explores the use of periphyton to ameliorate toxic properties of arsenic (As) to Labeo rohita and also assesses the human food safety aspects. Fish were introduced to arsenite [As(III)] contaminated water (0.3 and 3mg/L) along with periphyton. Biochemical, physiological and immunological parameters, including gene expression, were assessed after 30days of exposure. Periphyton incorporation significantly improved (p < 0.05) the adverse effects of As on respiration, NH3 excretion and brain AChE activity by reducing oxidative stress and As bioaccumulation. The presence of periphyton in As(III) exposed fish (3mg/L) increased the immune response (Immunoglobulin M and Complement C3) in the serum and the regulation of the respective immune genes in the anterior kidney was found to be similar to the control. A speciation study using LC-ICP-MS confirmed the high accumulation of As by periphyton (5.0-31.9μg/g) as arsenate [As (V)], resulting in a lower amount of As in fish muscle. The calculated human health risk indices, Target Hazard Quotient (THQ) and Target Cancer risk (TCR) indicate that fish grown in periphyton-treated water may lower the human health risks associated with As. The study signifies the importance of periphyton-based aquaculture systems in As contaminated regions for safe fish production with enhanced yield.

Trans-Cinnamaldehyde Primes More Robust Channel Catfish Immune Responses to Edwardsiella ictaluri Infection.

Infection with Edwardsiella ictaluri, a causative agent of enteric septicemia of catfish, threatens profitable catfish production through inventory losses. We previously demonstrated that trans-cinnamaldehyde (TC) enhances the survival of catfish following E. ictaluri infection. The present study was conducted to investigate catfish immune responses to TC feeding and E. ictaluri infection. The expression of 13 proinflammatory, innate, and adaptive immune-related genes was evaluated over time in two sets of experiments using real-time polymerase chain reaction (PCR). In the first experiment, catfish were fed a basal diet with or without TC supplementation, while in the second they were fed a TC-supplemented or normal diet followed by infection with E. ictaluri. The catfish group infected with E. ictaluri and fed a TC-diet showed significant changes in the expression of innate and adaptive immune-related genes compared to control group. At 21 and 28 days post-infection, recovered fish showed significant increases in the expression of IgM in the anterior kidney and spleen. These results suggest that the supplemental dietary intake of TC can improve the immune status of catfish via engaging innate and adaptive immune responses and the production of memory cells in immunocompetent tissues. Together, this study provides an important foundation for the potential application of TC as an antimicrobial alternative in aquaculture.

Collectin-K1 Plays a Role in the Clearance of Streptococcus agalactiae in Nile Tilapia (Oreochromis niloticus).

Collectin-K1 (CL-K1) is a multifunctional C-type lectin that has been identified as playing a crucial role in innate immunity. It can bind to carbohydrates on pathogens, leading to direct neutralization, agglutination, and/or opsonization, thereby inhibiting pathogenic infection. In this study, we investigated a homolog of CL-K1 (OnCL-K1) in Nile tilapia (Oreochromis niloticus) and its role in promoting the clearance of the pathogen Streptococcus agalactiae (S. agalactiae) and enhancing the antibacterial ability of the fish. Our analysis of bacterial load displayed that OnCL-K1 substantially reduced the amount of S. agalactiae in tissues of the liver, spleen, anterior kidney, and brain in Nile tilapia. Furthermore, examination of tissue sections revealed that OnCL-K1 effectively alleviated tissue damage and inflammatory response in the liver, anterior kidney, spleen, and brain tissue of tilapia following S. agalactiae infection. Additionally, OnCL-K1 was found to decrease the expression of the pro-inflammatory factor IL-6 and migration inhibitor MIF, while increasing the expression of anti-inflammatory factor IL-10 and chemokine IL-8 in the spleen, anterior kidney, and brain tissues of tilapia. Moreover, statistical analysis of survival rates demonstrated that OnCL-K1 significantly improved the survival rate of tilapia after infection, with a survival rate of 90%. Collectively, our findings suggest that OnCL-K1 plays a vital role in the innate immune defense of resisting bacterial infection in Nile tilapia. It promotes the removal of bacterial pathogens from the host, inhibits pathogen proliferation in vivo, reduces damage to host tissues caused by pathogens, and improves the survival rate of the host.

The Effect of Cynometra cauliflora Linn. Fruit Extract-Induced Congenital Anomalies on Chick Embryos

Background: Cynometra cauliflora is a traditional folk medicine that is used to treat a variety of unpleasant symptoms without adverse effects. Previous studies have shown that C. cauliflora fruit relieving morning sickness and nausea in pregnant women. However, no research has been performed on the effect of C. cauliflora fruit extract on birth anomalies. Objective: In this study, we aim to investigate the effects of C. cauliflora fruit extract on developing chick embryos. Materials and Methods: Three hundred fertilized eggs were divided into 2 groups; 1) the experimental groups received various doses of C. cauliflora fruit extract at 50, 100, 200, and 400 mg/kg, and 2) the control group received a 0.9 % normal saline solution. One hundred and fifty eggs at embryonic day-3 (ED-3) were observed for LC50, morphology, and histology, while the 150 eggs at embryonic day-6 (ED-6) were investigated for morphology and histology. Results: At a dosage of 291.70 mg/kg, the C. cauliflora fruit extract induced mortality in ED-3 by more than 50 %. Moreover, fruit extraction can also result in anomalies in ED-3 and ED-6, such as anterior neuropore openings, heart tubes, liver and kidney tissue defects. Conclusion: As a result, it was determined that tissue damage and organ malformation followed dose-dependent and all-or-none laws. Despite having a wide range of medical properties, C. cauliflora fruit extracts have been demonstrated to impair early growth and development and cause birth abnormalities in animal studies. This will allow further determination of regulatory criteria for medical use, particularly for pregnant women. HIGHLIGHTS First report of the cauliflora fruit extract induced embryonic lethality The cauliflora fruit extract induces embryonic organ and tissue anomalies following dose-dependent and all-or-none laws These experimental protocols are beneficial for teratogenesis studies GRAPHICAL ABSTRACT

Coinfection of channel catfish (Ictalurus punctatus) with virulent Aeromonas hydrophila and Flavobacterium covae exacerbates mortality.

Flavobacterium covae and virulent Aeromonas hydrophila are prevalent bacterial pathogens within the US catfish industry that can cause high mortality in production ponds. An assessment of invivo bacterial coinfection with virulent A. hydrophila (ML09-119) and F. covae (ALG-00-530) was conducted in juvenile channel catfish (Ictalurus punctatus). Catfish were divided into seven treatments: (1) mock control; (2) and (3) high and low doses of virulent A. hydrophila; (4) and (5) high and low doses of F. covae; (6) and (7) simultaneous challenge with high and low doses of virulent A. hydrophila and F. covae. In addition to the mortality assessment, anterior kidney and spleen were collected to evaluate immune gene expression, as well as quantify bacterial load by qPCR. At 96 h post-challenge (hpc), the high dose of virulent A. hydrophila infection (immersed in 2.3 × 107 CFU mL-1 ) resulted in cumulative percent mortality (CPM) of 28.3 ± 9.5%, while the high dose of F. covae (immersed in 5.2 × 106 CFU mL-1 ) yielded CPM of 23.3 ± 12.9%. When these pathogens were delivered in combination, CPM significantly increased for both the high- (98.3 ± 1.36%) and low-dose combinations (76.7 ± 17.05%) (p < .001). Lysozyme activity was found to be different at 24 and 48 hpc, with the high-dose vAh group demonstrating greater levels than unexposed control fish at each time point. Three proinflammatory cytokines (tnfα, il8, il1b) demonstrated increased expression levels at 48 hpc. These results demonstrate the additive effects on mortality when these two pathogens are combined. The synthesis of these mortality and health metrics advances our understanding of coinfections of these two important catfish pathogens and will aid fish health diagnosticians and channel catfish producers in developing therapeutants and prevention methods to control bacterial coinfections.

Antioxidant Defence in Labeo rohita to Biotic and Abiotic Stress: Insight from mRNA Expression, Molecular Characterization and Recombinant Protein-Based ELISA of Catalase, Glutathione Peroxidase, CuZn Superoxide Dismutase, and Glutathione S-Transferase.

Fish possess numerous enzymatic antioxidant systems as part of their innate immunity. These systems have been poorly studied in Labeo rohita (rohu). The present study characterized and investigated the role of antioxidant genes in the defence mechanisms against two types of stressors, including infection and ammonia stress. Four key genes associated with antioxidant activity-catalase, glutathione peroxidase, glutathione S-transferase, and CuZn superoxide dismutase were successfully cloned and sequenced. These genes were found to be expressed in different tissues and developmental stages of rohu. The expression levels of these antioxidant genes in the liver and anterior kidney tissues of rohu juveniles were modulated in response to bacterial infection (Aeromonas hydrophila), parasite infection (Argulus siamensis), poly I:C stimulation and ammonia stress. Additionally, the recombinant proteins derived from these genes exhibited significant antioxidant and antibacterial activities. These proteins also demonstrated a protective effect against A. hydrophila infection in rohu and had an immunomodulatory role. Furthermore, indirect ELISA assay systems were developed to measure these protein levels in healthy as well as A. hydrophila and ammonia-induced rohu serum. Overall, this study characterized and emphasised the importance of the antioxidant mechanism in rohu's defence against oxidative damage and microbial diseases.

Immunogenicity of extracellular proteins of Flavobacterium covae in Ictalurus punctatus

BackgroundColumnaris disease, instigated by Gram-negative Flavobacterium covae (F. covae), is a dreadful disease of many different genera of freshwater fish with alarmingly high mortality rates. Ictalurus punctatus (I. punctatus) is extremely prone to this disease. Despite the terrible implications of this disease, very little is known about how the host and the bacterium's virulence factor interact. Secreted extracellular proteins (SEPs) are key bacterial virulence agents that frequently provide immunogenicity. In a previous study, the major secreted extracellular proteins of F. covae were identified and their protective efficacy was verified in I. punctatus, and the results were published. In the current study, an endeavor has been made to examine the progressive shifts in I. punctatus immunity following immunization with F. covae SEPs in order to more thoroughly comprehend the host's intrinsic and acquired immune responses against F. covae infection. An enzyme-linked immunosorbent test (ELISA) is operated to analyze specific humoral immune responses against SEPs of F. covae. Moreover, the expressions of immune-associated genes that are related to inherent and acquired immunity were monitored in the anterior kidneys and spleens of the vaccinated I. punctatus.ResultsMarked rise in immunoglobulin M (IgM) levels were detected in the serum of I. punctatus vaccinated with different SEP formulations, including crude SEPs, heat-deactivated SEPs, and SEPs combined with adjuvant, at 14 and 21 days post-vaccination. At the same time points, a marked increase in the expression levels of clusters of differentiation (CD4-1 and CD8-β) and major histocompatibility complex (MHC-II) genes was detected in the vaccinated fish's anterior kidneys and spleens, while the highest levels of interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-α) up-regulations were detected in the immunized fish's anterior kidneys 14 days following vaccination.ConclusionsThis work revealed that SEPs are an effective basis for vaccination against F. covae infection, eliciting humoral and inherent immune responses of I. punctatus after SEPs immunization. Improvements are needed to further enhance the I. punctatus immunological responses to SEPs of F. covae since these antigens remain a prospective candidate for future optimization and clinical trials in aquaculture settings.

Unraveling the host-parasite interaction: immune response in Oreochromis mossambicus to Cymothoa eremita (Isopoda, Cymothoidae) infection.

The host immune response in Oreochromis mossambicus during natural infection with the isopod Cymothoa eremita was investigated. The naturally infected fishes were examined for enzyme profile, viz., respiratory burst activity (RBA), myeloperoxidase activity (MPO), and expression of immune-related genes, viz., toll-like receptor 22 (TLR 22), interleukin-1β (IL-1β), tumor necrosis factor α (TNF-α), complement component (C3), chemokine (CXCa), and β-actin in tissues of various organs (buccal cavity, gills and anterior kidney). Significant reduction (P < 0.05) in RBA and MPO was observed in the parasite-infected fishes when compared to the uninfected control fishes. In the buccal cavity, the expression of the immune-related genes was significantly (P < 0.05) upregulated, whereas all the genes except IL-1β were significantly (P < 0.05) upregulated in the anterior kidney. In the case of gill tissue, the expressed genes showed a varied type of regulation. The immunological responses in O. mossambicus during isopod infection have not been investigated in detail so far, and this is the first study unveiling such insights. Hence, this study will help to improve our molecular understanding of the host-immune response to parasitic isopod infection.

Transcriptomic and targeted immune transcript analyses confirm localized skin immune responses in Atlantic salmon towards the salmon louse.

Atlantic salmon (Salmo salar) are highly susceptible to infestations with the ectoparasite Lepeophtheirus salmonis, the salmon louse. Infestations elicit an immune response in the fish, but the response does not lead to parasite clearance, nor does it protect against subsequent infestations. It is, however, not known why the immune response is not adequate, possibly because the local response directly underneath the louse has been poorly evaluated. The present study describes the transcriptomic response by RNA sequencing of skin at the site of copepodid attachment. Analysing differentially expressed genes, 2864 were higher and 1357 were lower expressed at the louse attachment site compared to uninfested sites in the louse infested fish, while gene expression at uninfested sites were similar to uninfested control fish. The transcriptional patterns of selected immune genes were further detailed in three skin compartments/types: Whole skin, scales only and fin tissue. The elevation of pro-inflammatory cytokines and immune cell marker transcripts observed in whole skin and scale samples were not induced in fin, and a higher cytokine transcript level in scale samples suggest it can be used as a nonlethal sampling method to enhance selective breeding trials. Furthermore, the immune response was followed in both skin and anterior kidney as the infestation developed. Here, newly moulted preadult 1 stage lice induced a higher immune response than chalimi and adult lice. Overall, infestation with salmon louse induce a modest but early immune response with an elevation of mainly innate immune transcripts, with the response primarily localized to the site of attachment.

Co-exposure effects of lead and TiO2 nanoparticles in primary kidney cell culture from the freshwater fish Hoplias malabaricus

This study evaluated the effects of Lead (Pb) and titanium dioxide nanoparticles (TiO2 NPs) alone or in combination in anterior kidney macrophages of the freshwater fish Hoplias malabaricus, naïve or stimulated with 1 ng.mL-1 lipopolysaccharide (LPS). Pb (1 ×10-5 to 1 ×10-1 mg.mL-1) or TiO2 NPs (1.5 ×10-6 to 1.5 ×10-2 mg.mL-1) reduced cell viability despite LPS stimulation, especially Pb 10-1 mg.mL-1. In combination, lower concentrations of NPs intensified Pb-induced cell viability reduction while higher concentrations restored the cell viability independently of LPS stimulation. Basal and LPS- induced NO production was reduced by both TiO2 NPs and Pb isolated. The combination of both xenobiotics avoided this reduction of NO production by the isolated compounds at lower concentrations but the protective effect was lost as the concentrations increased. None xenobiotic increase DNA fragmentation. Therefore, at specific conditions, TiO2 NPs may have a protective effect over Pb toxicity, may also provide additional toxicity at higher concentrations.

Growth Retardation, Oxidative Stress, Immunosuppression, and Inflammatory Disturbances Induced by Herbicide Exposure of Catfish, Clarias gariepinus, and the Alleviation Effect of Dietary Wormwood, Artemisia cina

The present study evaluated the impact of chronic herbicide (oxyfluorfen; OXY) exposure on catfish, Clarias gariepinus, in terms of growth, hematobiochemical parameters, immune response, antioxidant- and immune-related gene expression, and resistance to monogenean parasites, Quadriacanthus aegypticus. In addition, the protective role of Wormwood, Artemisia cina (AC) against OXY exposure through diet inclusion was also analyzed. The catfish fingerlings were exposed to OXY (1.16 mg/L) for 60 days and fed diets without AC supplementation (control) and with 5% AC supplementation. The results demonstrated that exposure to OXY stunted growth; decreased survival, erythrograms and leukograms, serum protein, and acetylcholinesterase; and negatively altered the antioxidant status. On the contrary, AC supplementation significantly reduced OXY’s negative impacts on growth and hematological, biochemical, and antioxidant balance. In addition, exposure to OXY markedly increased levels of biomarkers of hepatorenal damage, stress indicators, and DNA damage, which were alleviated with AC supplementation. OXY exposure induced immunosuppression manifested by a decrease in lysozyme activities, complement c3, nitric oxide levels, and phagocytic activity. Furthermore, exposure to OXY negatively regulated the expression of immune-antioxidant genes (CAT, GPX1, SOD1, GST, and TGF-Β1). However, it upregulated the expression of CYP1a, IL-1β, and TNF-α in the liver, anterior kidney, and intestine of C. gariepinus. Meanwhile, the addition of AC to the OXY-exposed fish diets notably restored immune components and remedied the altered immune-related gene expressions. Likewise, the AC supplementation significantly alleviated the OXY-induced reduction in the fish survival rate after Q. aegypticus challenge. Accordingly, AC dietary supplementation in catfish diets could alleviate the negative impact of exposure to OXY on growth performance, physiological status, and some immune-antioxidant-related gene expression.

Streptococcus agalactiae Serotype VII, an Emerging Pathogen Affecting Snakeskin Gourami (Trichogaster pectoralis) in Intensive Farming

Snakeskin gourami (Trichogaster pectoralis) is a freshwater fish species that is being increasingly cultivated in Southeast Asia. The expansion of farms and intensive farming practices has led to the unexplained mortality of snakeskin gourami and tremendous economic losses in many farms. We investigated the unusual mortality of snakeskin gourami at 22 farms in Central Thailand. The moribund fish showed darkened skin, erratic swimming, exophthalmos, and haemorrhaging around the eyeballs, with cumulative mortality between 20% and 45%. Our necropsy findings revealed an enlarged liver and anterior kidney, splenomegaly, haemorrhage in most internal organs, pericarditis, and brain congestion. Histopathology revealed haemorrhaging and congestion of the blood vessels in the liver with infiltration of lymphocytes, enlarged blood vessels with mononuclear and lymphocyte infiltration in the meninges, and cerebral parenchyma were observed. Severe necrotising and suppurative pericarditis with myocardial infarction were found. Epidemiological studies and laboratory diagnosis revealed that Streptococcus agalactiae was predominantly isolated from the moribund fish. Laboratory investigations of the representative 33 isolates of S. agalactiae using mass spectrometry, multiplex polymerase chain reaction assay, pulse-gel electrophoresis, and serotyping suggested that all the isolates were S. agalactiae serotype VII, which is different from the serotype III isolated from diseased tilapia in Thailand. An experimental challenge using three representative isolates of S. agalactiae on snakeskin gourami caused clinical signs, gross lesions, and pathological changes, with high mortality exceeding 60%, which is similar to the mortality in most natural infections. Moreover, S. agalactiae was recovered from the spleen, kidneys, and liver of all the challenged fish. Taken together, this study provides important information that S. agalactiae serotype VII is virulent in snakeskin gourami and can potentially spread among these fish in fish farms. Appropriate preventive measures and the control of animal movements should thus be implemented.

Application of a Lipopolysaccharide (LPS)-Stimulated Mitogenesis Assay in Smallmouth Bass (Micropterus dolomieu) to Augment Wild Fish Health Studies

The utility of a functional immune assay for smallmouth bass (Micropterus dolomieu) lymphocyte mitogenesis was evaluated. Wild populations in the Potomac River have faced disease and mortality with immunosuppression from exposure to chemical contaminants a suspected component. However, a validated set of immune parameters to screen for immunosuppression in wild fish populations is not available. Prior to use in ecotoxicology studies, ancillary factors influencing the mitogenic response need to be understood. The assay was field-tested with fish collected from three sites in West Virginia as part of health assessments occurring in spring (pre-spawn; April–May) and fall (recrudescence; October–November). Anterior kidney leukocytes were exposed to lipopolysaccharide (LPS) from E.coli O111:B4 or mitogen-free media and proliferation was measured using imaging flow cytometry with advanced machine learning to distinguish lymphocytes. An anti-smallmouth bass IgM monoclonal antibody was used to identify IgM+ lymphocytes. Lymphocyte mitogenesis, or proliferative responses, varied by site and season and positively and negatively correlated with factors such as sex, age, tissue parasites, and macrophage aggregates. Background proliferation of IgM− lymphocytes was negatively correlated to LPS-induced proliferation in both seasons at all sites, but only in spring for IgM+ lymphocytes. The results demonstrate that many factors, in addition to chemical contaminants, may influence lymphocyte proliferation.

Chromosome Mapping of Repetitive DNAs in the Picasso Triggerfish (Rhinecanthus aculeatus (Linnaeus, 1758)) in Family Balistidae by Classical and Molecular Cytogenetic Techniques

This work presents the cytogenetic analysis conducted on the Picasso triggerfish (Rhinecanthus aculeatus (Linnaeus, 1758)) from Thailand. Mitotic chromosomes were prepared from the anterior kidney. The cell suspensions were harvested by in vivo colchicine treatment. The present study includes the chromosomal investigation on R. aculeatus, using conventional (Giemsa staining, Ag-NOR and C-banding) and molecular approaches (in situ mapping of five different repetitive DNA classes including 18S rDNA, 5S rDNA, (CA)15, (GA)15 and (CAA)10 as markers.) The results showed that R. aculeatus has karyotypes formed exclusively by telocentric chromosomes (44t; NF=44). The C-positive heterochromatic blocks are preferentially located in the centromeric and telomeric regions of some chromosomal pairs. The Ag-NOR sites occupy the interstitial position of the long arms of the largest telocentric pair (pair 1). The exclusive location of the major ribosomal sites in these pairs was confirmed by hybridization with 18S rDNA probes. However, the 5S rDNA genes are not located on 18S rDNA-bearing chromosomes, but instead located exclusively in the subcentromeric region of pair 4. The mapping of (CA)15, (GA)15 and (CAA)10 microsatellites are sparsely dispersed along all the chromosomes. The karyotype formula of R. aculeatus is 2n (44) = 44t.

Cellular composition of the kidneys of the common carp after the use of cellobacterin-T under conditions of anthropogenic pressure

The study presents the data on the cellular composition of the kidneys of the common carp against the background of the use of the probiotic Celloblacterin-T. The experiment was planned and carried out at a specialized fish farm in Voronezh region. Specimens of the common carp aged 1 year and weighing in the range of 250-300 grams were selected for the experiment. As part of the baseline study, kidneys were taken from fish (n=5) for cytological studies. After that, the fish were divided into 2 groups: carps from the control (n=8000) and experimental (n=8000) ponds. The fish from the control pond received the main diet, and the fish from the experimental pond received the probiotic drug Cellobacterin-T (with the main diet), calculated at a dose of 2 kg/t of feed. Then a test fishing was carried out, during which kidneys were taken from the specimens of both ponds (n=8 from each) for cytological studies. They were conducted 14 days after the beginning of using the probiotic and 42 days after the beginning (12 days after the end of using the probiotic). In the baseline study, we revealed inhibition of the lymphoid lineage of the anterior kidney, an inflammatory neutrophilic reaction caused by the effect of heavy metals, as evidenced by a low number of lymphocytes (16.5%) and a high number of neutrophils and their precursors (10.5%). Against the background of the use of Cellobacterin-T, activation of lymphocytes was observed, their number increased by 1.9 times relative to the baseline study. The number of neutrophils and their precursors was reduced by 3 times. After discontinuation of the drug, the results of the experimental group were close to the results of the baseline study.

Role of computed tomography imaging in the diagnosis of blunt and penetrating abdominal trauma injuries

Background: The present study was conducted with the primary aim of evaluating the role of computed tomography imaging in the assessment of blunt abdominal trauma (BAT) and penetrating abdominal trauma injuries. Methods: This prospective study comprised 53 patients with BAT and PAT injuries referred from the emergency department to the department of radio-diagnosis for ultrasonography and CT. Multidetector computed tomography scans were performed on GE Bright Speed Elite 16 slice CT Scanner. The various injuries seen on the CT images were grouped and examined based on the injury site and the organs involved. Results: The majority of the study subjects, i.e., 17/53 (32.08%) belonged age group of 20-30 yrs. Out of the 53 study subjects, 48 had BAT and 5 had PAT. The majority of study subjects who had BAT, i.e., 26/48 RTA, was the cause of trauma, and 5 out of 5 who had PAT assault was the cause of trauma. The most common organ injured was the spleen (18), followed by the liver (17), bowel (15), Anterior abdominal wall (6), pancreas (1), Kidney (1), Uterus (2), Multiple organs (1) and Hemoperitoneum without solid organ injury (1). CT showed 100% accuracy in the diagnosis of abdominal organ injuries. Conclusions: CT imaging plays a pivotal role in the rapid identification of life-threatening injuries and is helpful for prompt initiation of appropriate care, which leads to an increase in the chance of survival of patients with BAT and PAT injuries.

Growth Performance, Survival, Blood Chemistry, and Immune Gene Expression of Channel Catfish (Ictalurus punctatus) Fed Probiotic-Supplemented Diets.

The channel catfish (Ictalurus punctatus) farming industry is the largest and one of the oldest aquaculture industries in the United States. Despite being an established industry, production issues stemming from disease outbreaks remain problematic for producers. Supplementing fish diets with probiotics to enhance the immune system and growth potential is one approach to mitigating disease. Although considerable laboratory data demonstrate efficacy, these results do not always translate to natural modes of disease transmission. Hence, the present work was conducted in the laboratory but incorporated flow-through water from large catfish pond production systems, allowing for natural exposure to pathogens. Two feeding trials were conducted in an 18-tank aquaria system housing two different sizes, 34.8 ± 12.5 g and 0.36 ± 0.03 g, of channel catfish. Channel catfish in the first trial were fed three experimental diets over six weeks. Commercial diets were top-coated with two selected spore-forming Bacillus spp. probiotics, Bacillus velezensis AP193 (1 × 106 CFU g-1) and BiOWiSH (3.6 × 104 CFU g-1), or a basal diet that contained no dietary additive. In the second eight-week trial, diets were top-coated with BiOWiSH at three concentrations (1.8, 3.6, and 7.3 × 104 CFU g-1), along with one basal diet (no probiotic). At the completion of these studies, growth performance, survival, hematocrit, blood chemistry, and immune expression of interleukin 1β (il1β), tumor necrosis factor-alpha (tnf-α), interleukin-8 (il8), transforming-growth factor β1 (tgf-β1), and toll-like receptor 9 (tlr9) were evaluated using qPCR. Trial results revealed no differences (p &gt; 0.05) among treatments concerning growth, survival, or hematological parameters. For immune gene expression, interesting trends were discerned, with substantial downregulation observed in B. velezensis AP193-fed fish for il1β, tnf-α, and tlr9 expression within splenic tissue, compared to that of the basal and BiOWiSH diets (p &lt; 0.05). However, the results were not statistically significant for anterior kidney tissue in the first trial. In the second trial, varied levels of probiotic inclusion revealed no significant impact of BiOWiSH's products on the expression of il1β, tnf-α, il8, and tgf-β1 in both spleen and kidney tissue at any rate of probiotic inclusion (p &gt; 0.05). Based on these findings, more research on utilizing probiotics in flow-through systems with natural infection conditions is crucial to ensure consistency from a controlled laboratory scale to real-world practices.

Transperitoneal laparoscopic and robot-assisted resection of anterior and posterior kidney tumors

Background. Laparoscopic and robot-assisted resection of the posterior kidney tumors can be performed through retroperitoneal and transperitoneal accesses. Supporters of retroperitoneal partial nephrectomy point to better results for neoplasms in this location. However, most surgeons use transperitoneal access.Aim. To perform comparative analysis of the results of laparoscopic and robot-assisted partial resection of anterior and posterior renal neoplasms.Materials and methods. The results of transperitoneal minimally invasive partial nephrectomy in 156 patients (86 (55.1 %) men and 70 (44.9 %) women) were analyzed. Anterior tumors (1st group) were diagnosed in 96 (61.5 %), posterior (2nd group) in 60 (38.5 %) patients. In the 1st group, mean tumor size was 3.4 ± 1.6 cm; in the 2nd group, 3.2 ± 1.4 cm. In the 1st group, laparoscopic partial nephrectomy was performed in 70 (72.9 %), robot-assisted in 26 (27.1 %) patients; in the 2nd group in 40 (66.7 %) and 20 (33.3 %) patients, respectively. Comparative analysis of operating time, warm ischemia time, complications and glomerular filtration rate was performed.Results. No conversions into nephrectomy or open interventions were performed. Time of laparoscopic partial nephrectomy in the 1st group was shorter than in the 2nd: 108.6 min versus 120.5 min (р &gt;0.05). In robot-assisted partial nephrectomy it was 90.5 and 112.6 min (р &gt;0.05), respectively. For laparoscopic partial nephrectomy, mean warm ischemia time in the 1st group was 15.0 min, in the 2nd group it was 17.5 min (р &gt;0.05); in robot-assisted operation it was 13.2 and 16.0 min (р &gt;0.05), respectively. Blood loss volume was lower in the 1st group compared to the 2nd: 125 ml versus 230 ml (р &lt;0.05). In all patients, renal cell carcinoma was diagnosed; Т1а tumors were predominant (in the 1st and 2nd group, 87.5 and 88.3 %, respectively; р &gt;0.05). Postoperative complications in transperitoneal resection of anterior tumors were observed less frequently: 16.7 % versus 20.0 % (р &gt;0.05). Grade I complications per the Clavien classification in the 1st and 2nd group were observed in 8 (8.3 %) and 6 (10.0 %) patients, grade II in 6 (6.3 %) and 5 (8.3 %) patients, grade III in 2 (2.1 %) and 1 (1.7 %) patients. After resection, urinary leakage through the drain was observed in 2 (2.1 %) patients of the 1st group and 1 (1.7 %) patient of the 2nd group; they underwent drainage of the upper urinary tract using a stent. Blood transfusion in the postoperative period was performed in 2 patients from each group (2.1 and 3.3 %, respectively). In the 1st group, glomerular filtration rate before and after surgery was 68 and 56 ml/min/1.73 m2, in the 2nd group 70.2 and 58.2 ml/min/1.73 m2, respectively; no significant difference was observed (р &gt;0.05).Conclusion. Transperitoneal access allows to safely perform minimally invasive resection of the anterior and posterior kidney tumors with results comparable to the perioperative results of retroperitoneal partial nephrectomy.

Teleost fish IgM+ plasma-like cells possess IgM-secreting, phagocytic, and antigen-presenting capacities.

Plasma cells are terminally differentiated antibody-secreting B lymphocytes that contribute to humoral immunity by producing large numbers of antibodies. Increasing evidence suggests that teleost fish B cells share certain characteristics with mammalian B1 B cells, including antibody-secreting, phagocytic, and antigen-presenting capacities. However, the difference between mature B cells and plasma cells remains unclear. In this study, we found that, based on their light-scattering characteristics, tilapia anterior kidney (AK) leukocytes can be categorized into two IgM+ B-cell subsets: the lymphoid (L) gate and granulocyte–monocyte/macrophage (G-M) subsets. G-M gate cells are more numerous than L-gate cells and have higher mean fluorescence, but lower forward scatter and side scatter. We analyzed the morphological and ultrastructural features of sorted IgM+ cells and found that L-gate IgM+ cells have a high nucleus–cytoplasm ratio and lymphocyte-like morphology, whereas G-M gate IgM+ cells have a small nucleus, more abundant endoplasmic reticulum, and a larger number of mitochondria, and have a plasma cell-like or macrophage-like morphology. To further characterize the cell types, we examined the specific patterns of expression of B-cell- and T-cell-related genes. We found that B-cell-specific genes were expressed by both L-gate and G-M gate IgM+ cells, and that G-M gate IgM+ cells secreted extremely high levels of IgM. However, T-cell-related genes were highly expressed only in L-gate IgM– cells. These results suggest that G-M gate IgM+ cells are similar to plasma-like cells, with high antibody-secreting capacity. Given that G-M gate cells include the granulocyte, monocyte, and macrophage cell types, but not B cells, monocyte/macrophage markers were used to investigate the cell types further. A macrophage receptor with a collagenous structure was frequently observed, and macrophage-expressed gene-1 was highly expressed, in the G-M gate IgM+ cells. Phagocytic capacity, as determined by ingestion of beads or bacteria, was significantly higher in G-M gate IgM+ cells than in L-gate IgM+ cells, as was antigen-processing capacity. Our findings show that tilapia AK leukocytes can be divided into two IgM+ B-cell subsets and that G-M gate IgM+ cells resemble plasma-like cells, having high antibody-secreting, phagocytic, and antigen-presenting capacities. Thus, this study increases our understanding of the functions of teleost fish plasma-like cells.

A simple method to evaluate the toxic effects of Prorocentrum lima extracts to fish (sea bass) kidney cells

The diarrhetic shellfish toxins (DSTs) okadaic acid (OA) and its analogues — the dinophysistoxins (DTXs) — are produced by dinoflagellates such as Prorocentrum lima and can bioaccumulate in filter-feeding organisms as they are transferred through the food web. Although there is no assessment of the harmful effects of these toxins on the fish's immune system, this study developed a primary culture protocol for kidney cells from marine fish Centropomus parallelus and evaluated the immunotoxic effects to P. lima extracts containing DSTs. The cells were obtained by mechanical dissociation, segregated with Percoll gradient, and incubated for 24 h at 28 °C in a Leibovitz culture medium supplemented with 2% fetal bovine serum and antibiotics. The exposed cells were evaluated in flow cytometry using the CD54 PE antibody. We obtained >5.0 × 106 viable cells per 1.0 g of tissue that exhibited no cell differentiation. Exposure to 1.2 or 12 ng DST mL−1 stimulated the immune system activation and increased the proportion of activated macrophages and monocytes in 48 to 52% and in 127 to 146%, respectively. The protocol proved to be an alternative tool to assess the immunotoxic effects of DST exposure on fish's anterior kidney cells.