Renal Hematopoietic Tissue Research Articles

Perfluorooctanoic Acid Promotes Recruitment and Exocytosis of Rodlet Cells in the Renal Hematopoietic Tissue of Common Carp.

Per- and polyfluoroalkyl substances (PFAS) are ubiquitous environmental contaminants, with perfluorooctanoic acid (PFOA) being a prominent member. PFOA poses a risk to aquatic ecosystems and human health due to its presence in water, environmental persistence, and bioaccumulation. Since rodlet cells (RCs) have emerged as potential biomarkers for chemical stressors, this study aimed to investigate the effects of sub-chronic PFOA exposure on RCs in the renal hematopoietic tissue of common carp. Three groups of fish were used: an unexposed control group and two groups exposed to environmentally relevant (200 ng L-1) and elevated (2 mg L-1) PFOA concentrations. Light and transmission electron microscopy were employed to assess RCs' distribution patterns and exocytosis, while biometry quantified RCs in the hematopoietic tissue. The results showed that, even at environmentally relevant concentrations, PFOA significantly influenced RCs' distribution patterns, leading to increased occurrence and cluster formation, as well as heightened exocytosis activity. This research highlights PFOA's immunotoxicity in fish and suggests the potential of RCs as sentinel cells in the immunological response to environmental contaminants. These findings enhance our understanding of PFAS toxicity and emphasise the importance of monitoring their impact on fish as representative vertebrates and reliable animal models.

Investigation of laboratory methods for characterization of aquatic viruses in fish infected experimentally with infectious salmon anemia virus.

Rapid growth in aquaculture has resulted in high-density production systems in ecologically and geographically novel conditions in which the emergence of diseases is inevitable. Well-characterized methods for detection and surveillance of infectious diseases are vital for rapid identification, response, and recovery to protect economic and food security. We implemented a proof-of-concept approach for virus detection using a known high-consequence fish pathogen, infectious salmon anemia virus (ISAV), as the archetypal pathogen. In fish infected with ISAV, we integrated histopathology, virus isolation, whole-genome sequencing (WGS), electron microscopy (EM), in situ hybridization (ISH), and reverse transcription real-time PCR (RT-rtPCR). Fresh-frozen and formalin-fixed tissues were collected from virus-infected, control, and sham-infected Atlantic salmon (Salmo salar). Microscopic differences were not evident between uninfected and infected fish. Viral cytopathic effect was observed in cell cultures inoculated with fresh-frozen tissue homogenates from 3 of 3 ISAV-infected and 0 of 4 uninfected or sham-infected fish. The ISAV genome was detected by shotgun metagenomics in RNA extracted from the medium from 3 of 3 inoculated cell cultures, 3 of 3 infected fish, and 0 of 4 uninfected or sham-infected fish, yielding sufficient coverage for de novo assembly. An ISH probe against ISAV revealed ISAV genome in multiple organs, with abundance in renal hematopoietic tissue. Virus was detected by RT-rtPCR in gill, heart, kidney, liver, and spleen. EM and metagenomic WGS from tissues were challenging and unsuccessful. Our proof-of-concept methodology has promise for detection and characterization of unknown aquatic pathogens and also highlights some associated methodology challenges that require additional investigation.

Multi-tool diagnosis of an outbreak of ranavirosis in amphibian tadpoles in the Canadian boreal forest.

Investigation of mortalities in isolated wild amphibian populations presents diagnostic difficulties that can hinder reaching a definitive diagnosis for the cause of death. Disease can only be diagnosed when pathogen presence (e.g. detection by PCR) is linked to tissue lesions (histopathology) in the host. We report a 2-site outbreak of ranavirosis in wild anuran tadpoles in the boreal forest of Wood Buffalo National Park, Canada, diagnosed by histologic and molecular techniques. Mortalities occurred in wood frog Rana sylvatica tadpoles and boreal chorus frog Pseudacris maculata tadpoles. Lack of mortality in sympatric Canadian toad Bufo (Anaxyrus) hemiophrys tadpoles suggested lower disease susceptibility in this species. In the former 2 species, ranavirosis was diagnosed based on consistent histopathology, immunohistochemistry (IHC), in situ hybridization (ISH), and quantitative PCR results. The most common histopathologic lesion present in wood and boreal chorus frog tadpoles was necrosis of the skin, oral mucosa, renal tubular epithelium, renal hematopoietic tissue, and branchial epithelium. Mild hepatic and pancreatic necrosis and rare intracytoplasmic inclusion bodies in hepatocytes were less common. Skeletal and connective tissues in budding limbs often had multifocal to coalescing necrosis and were intensely positive for ranavirus, with IHC staining even in areas where no obvious necrosis could be observed. Abundant IHC and ISH staining in actively growing tissues support a link between disease emergence and amphibian developmental stage. Our findings provide a definitive diagnosis of ranavirosis in free-living amphibians and highlight the effectiveness of multi-tool approaches to mortality investigation and elucidation of pathogenesis of ranavirosis in wild amphibians.

Citrobacter freundii infection in silver catfish (Rhamdia quelen): Hematological and histological alterations

Citrobacter freundii is a fish pathogen known for its ability to cause injury and high mortality. There have been no studies reporting the effect of this bacterium on hematological parameters and internal organ histology in silver catfish (Rhamdia quelen). Therefore, the aim of this study is to evaluate the hematological and histopathological effects of an experimentally induced C. freundii infection in silver catfish. Twenty fish were divided into healthy and infected groups. The fish of the infected group were inoculated intramuscularly with 100 μL of bacterial suspension (6.4 × 108 CFU mL−1), while healthy control animals received 100 μL of sterile saline. On day 18 post-infection, blood and tissues (cephalic kidneys, livers, and spleens) were collected for histological analysis. The infected animals presented high mortality, as well as hematological and histological changes. In relation to hematology, the infected fish presented aregenerative anemia, protein loss, leukopenia with neutropenia, lymphocytosis, and leukoblastosis. Regarding histology, there was liver degeneration, decrease in the amount of renal hematopoietic tissue, and the presence of melanomacrophage centers (MMCs) in the spleen and cephalic kidney of infected fish. In summary, these alterations may contribute to disease pathophysiology, contributing to high mortality of affected fish.

Diclofenac affects kidney histology in the three-spined stickleback (Gasterosteus aculeatus) at low μg/L concentrations

Diclofenac, a commonly used non-steroidal anti-inflammatory drug, is considered for regulation under the European water framework directive. This is because effects on fish have been reported at concentrations around those regularly found in treated sewage effluents (∼1μg/L). However, a recent publication reports no effects on fish at 320μg/L. In this study, three-spined sticklebacks (Gasterosteus aculeatus) were exposed to 0, 4.6, 22, 82 and 271μg/L diclofenac in flow-through systems for 28days using triplicate aquaria per concentration. At the highest concentration, significant mortalities were observed already after 21days (no mortalities found up to 22μg/L). Histological analysis revealed a significant increase in the proportion of renal hematopoietic tissue (renal hematopoietic hyperplasia) after 28days at the lowest concentration and at all higher concentrations, following a clear dose-response pattern. Skin ulcerations of the jaw were noted by macroscopic observations, primarily at the two highest concentrations. No histological changes were observed in the liver. There was an increase in the relative hepatic mRNA levels of c7 (complement component 7), a gene involved in the innate immune system, at 22μg/L and at all higher concentrations, again following a clear dose-response. The bioconcentration factor was stable across concentrations, but lower than reported for rainbow trout, suggesting lower internal exposure to the drug in the stickleback. In conclusion, this study demonstrates that diclofenac causes histological changes in the three-spined stickleback at low μg/L concentrations, which cause concern for fish populations exposed to treated sewage effluents.

Ontogeny of American paddlefish lymphoid tissues

The temporal and spatial distribution of American paddlefish Polyodon spathula immune cell populations was determined using enzyme cytochemistry and immunohistochemistry. Monocytes and macrophages were present in the renal haematopoietic tissue, spleen, meningeal myeloid tissue, cardiac myeloid tissue and lamina propria of the spiral valve at 7 days post‐hatch (dph). Immature lymphocytes were present in the renal haematopoietic tissue, spleen, meningeal myeloid tissue, cardiac myeloid tissue, thymus and lamina propria of the spiral valve at 7 dph. Type A lymphocytes (T‐cell like) were demonstrated in the thymus by 21 dph. Type B immunoglobulin positive lymphocytes (B‐cell like) were present in the renal haematopoietic tissue, cardiac myeloid tissue and lamina propria of the spiral valve by 7 dph, the thymus at 21 dph, the spleen by 56 dph, and were not observed in the meningeal myeloid tissue of paddlefish aged 7–28 dph. Granulocytes were present in the renal haematopoietic tissue, thymus, spleen, meningeal myeloid tissue, cardiac myeloid tissue and lamina propria of the spiral valve by 7 dph. The spleens in 7–28 dph fish were predominately red pulp. Differentiation of leukocytic and erythrocytic compartments (white and red pulp, respectively) was not apparent in the spleen until 56 dph. Remarkable thymic cortical and medullary differentiation was not yet present at 28 dph, and the thymus was not sampled at 56, 84 or 112 dph. The cardiac myeloid tissue was not developed until 56 dph. Peyer’s patches were present in the lamina propria by 56 dph. Paddlefish lympho‐myeloid structures are therefore slow to develop, and vaccination procedures should be performed at 2–4 months post‐hatch.

American paddlefish leukocytes demonstrate mammalian‐like cytochemical staining characteristics in lymphoid tissues

American paddlefish Polyodon spathula leukocytes demonstrated cytoplasmic staining patterns very similar to mammalian leukocytes when stained with acid phosphatase, α‐naphthyl butyrate esterase and β‐glucuronidase. American paddlefish monocytes, lymphocytes and granulocytes stained positive for acid phosphatase. Monocytes stained positive for α‐naphthyl butyrate esterase. Lymphocytes that stained positive for α‐naphthyl butyrate esterase were designated type A. Lymphocytes that stained positive with antibodies to the L chain of white sturgeon Acipenser transmontanus immunoglobulin (Ig) were designated type B. Type A and type B lymphocytes stained positive for β‐glucuronidase. All leukocytes observed were negative for Sudan Black B. Monocytes, lymphocytes and granulocytes were present in the renal haematopoietic tissue, spleen, thymus, pericardial myeloid tissue, lamina propria of the spiral valve, and in meningeal myeloid tissue located dorsal to the brain, at the base of the brain and around the notochord. Peyer's patches were present in the gut. Morphological characteristics of leukocytes stained with Wright's and haematoxylin and eosin and appeared very similar to those of other fish species.

Ontogeny of channel catfish lymphoid organs

Previously, we showed that catfish could not mount a detectable antibody response after bacterial exposure until 21 days post-hatch (ph). In order to evaluate the changes associated with the development of a functional humoral response, we evaluated the temporal and spatial distribution of immune cell populations in developing catfish. Cells functioning in nonspecific immunity were present in the renal hematopoietic tissue (rht) and thymus at hatch and in the spleen by day 3 ph. Immunoglobulin (Ig) positive lymphocytes were first detected on day 7, 10, and 14 in the rht, thymus and spleen, respectively. Mature thymocytes were first detected on day 10 ph. Distinct thymic regionalization and splenic lymphoid tissue organization were not observed until day 21 ph. We suggest that the reason for a lack of antibody production until day 21 ph is the poor organization of secondary lymphoid tissue until that age.

Pathogenesis of Iridovirus: in vitro Influence on Macrophage Activity and Cytokine-Like Protein Production in Fish

Pathogenesis of Iridovirus: in vitro Influence on Macrophage Activity and Cytokine-Like Protein Production in Fish

Differential cytochemical staining characteristics of channel catfish leukocytes identify cell populations in lymphoid organs

This is one of the first characterizations of channel catfish ( Ictalurus punctatus) leukocytes by enzyme cytochemistry. Leukocytes demonstrated cytoplasmic staining patterns very similar to mammalian leukocytes when stained with acid phosphatase, α-naphthyl butyrate esterase, β-glucuronidase, α-naphthyl acetate esterase, Sudan Black B and anti-immunoglobulin specific immunohistochemistry. Lymphocytes, monocytes, macrophages, neutrophils, and surface immunoglobulin positive (surface Ig+) cells were present in channel catfish renal hematopoietic tissue and spleen and demonstrated distinctive cytoplasmic foci staining patterns, cytoplasmic blushing or cell membrane staining. Monocytes, macrophages, lymphocytes and surface Ig+ cells were present in the thymus. Thymic and splenic cellular organization appeared very similar to these same mammalian tissues. In the thymus, acid phosphatase positive cells were distributed throughout the parenchyma, while α-naphthyl butyrate esterase and β-glucuronidase positive cells were concentrated in the cortex and the medulla, respectively. Surface immunoglobulin positive cells occurred in the cortex. In the spleen, acid phosphatase positive cells were scattered throughout the parenchyma, while α-naphthyl butyrate esterase positive cells were scattered throughout the parenchyma and adjacent to splenic arterioles. β-glucuronidase and surface immunoglobulin positive cells were restricted to immediately adjacent to splenic arterioles. Sudan Black B positive cells were scattered throughout the parenchyma, while α-naphthyl acetate esterase positive cells occurred adjacent to peri-arteriole lymphoid sheaths and appear very similar to mammalian metallophils.

Early hematopoiesis and developing lymphoid organs in the zebrafish.

In zebrafish, the transparent and rapidly developing embryo and the potential for genetic screening offer a unique opportunity to investigate the early development of the vertebrate immune system. Here we describe the initial appearance of various blood lineages and the nature of accumulating hematopoietic tissue in the thymus and kidney, the main lymphoid organs of adult teleosts. The ultrastructure of the first site of hematopoiesis, the intermediate cell mass (ICM), is described from the 5-somite stage, about 11.5 hours post-fertilization (hpf) until 24 hpf. The ICM gives rise to the primitive erythroid lineage, which accounts for all circulating erythrocytes for the first 4 days pf. From 24 to 72 hpf, a few developing granulocytes are seen close to the yolk sac walls and in the caudal axial vein. The heart, previously proposed as an early blood-forming organ in zebrafish, did not contain hematopoietic cells. The thymic primordium, consisting of two layers of epithelial cells, appears at 60 hpf. By 65 hpf, it is colonized by immature lymphoblasts. The thymus gradually accumulates lymphocytes, and the lymphocytes and epithelial cells progressively differentiate for 3 weeks pf. At 96 hr, the pronephros contains hematopoietic cells, mainly developing erythrocytes and granulocytes. The amount of renal hematopoietic tissue increases rapidly; however, lymphocytes were not detected until 3 weeks pf.

Early hematopoiesis and developing lymphoid organs in the zebrafish

In zebrafish, the transparent and rapidly developing embryo and the potential for genetic screening offer a unique opportunity to investigate the early development of the vertebrate immune system. Here we describe the initial appearance of various blood lineages and the nature of accumulating hematopoietic tissue in the thymus and kidney, the main lymphoid organs of adult teleosts. The ultrastructure of the first site of hematopoiesis, the intermediate cell mass (ICM), is described from the 5-somite stage, about 11.5 hours post-fertilization (hpf) until 24 hpf. The ICM gives rise to the primitive erythroid lineage, which accounts for all circulating erythrocytes for the first 4 days pf. From 24 to 72 hpf, a few developing granulocytes are seen close to the yolk sac walls and in the caudal axial vein. The heart, previously proposed as an early blood-forming organ in zebrafish, did not contain hematopoietic cells. The thymic primordium, consisting of two layers of epithelial cells, appears at 60 hpf. By 65 hpf, it is colonized by immature lymphoblasts. The thymus gradually accumulates lymphocytes, and the lymphocytes and epithelial cells progressively differentiate for 3 weeks pf. At 96 hr, the pronephros contains hematopoietic cells, mainly developing erythrocytes and granulocytes. The amount of renal hematopoietic tissue increases rapidly; however, lymphocytes were not detected until 3 weeks pf. Dev Dyn 1999;214:323–336. © 1999 Wiley-Liss, Inc.

Tissue distribution and structure of barrier cells in the hematopoietic and lymphoid organs of salmonids.

Barrier cells have been recognized as a discrete group of fibroblastic- or myofibroblastic-like cells located in the lymphoid and hematopoietic organs of mammals. This paper reports the results of a morphological study of the main lymphoid organs of three salmonid species, in which cells structurally similar to the mammalian barrier cells were observed in healthy animals. The spleen, kidney, and thymus of fingerlings of rainbow trout, Oncorhynchus mykiss, and Coho salmon, Oncorhynchus kisutch, and of adult brown trout, Salmo trutta fario, were processed for electron microscopy study using various fixation methods. Semithin sections were used for the Periodic Acid-Schiff (PAS) staining technique, and for the demonstration of the endogenous peroxidase activity. The kidney and spleen of all the species contained a variable, but usually low, number of electron-dense, elongated, and branched cells, ultrastructurally similar to the mammalian barrier cells. They also occurred in the thymus of some brown trout and Coho salmon, but not of rainbow trout. The electron density of this cell type was present after the various types of fixation procedures. They show numerous ribosomes, well-developed secretory organelles, electron-clear vesicles, large granules, and microfilaments. In all the salmonid species, barriers cells were positive for PAS staining and for endogenous peroxidase activity. A small number of barrier cells were in mitosis. In the different organs barrier cells appeared as isolated cells, or forming syncytial networks. They were found lining the blood sinusoids of the splenic red pulp and of the renal hematopoietic tissue, in the periellipsoidal sheaths, and closely associated with erythropoietic and plasmacytopoietic foci. Our results demonstrate the presence of cells closely resembling mammalian barrier cells in the hematopoietic and lymphoid organs of salmonids. The structure and tissue distribution of the salmonid barrier cells are discussed in relation to the functional roles described for this cell type in mammals.

Epizootic haematopoietic necrosis, a new viral disease in redfin perch, Perca fluviatilis L., in Australia

Abstract. An epizootic disease in redfin perch, Perca fluviatilis L., in north‐east Victoria, Australia, was found to be caused by a recently described iridovirus. The acute disease is characterized by necrosis of the renal haematopoietic tissue, liver and spleen, with variable involvement of the pancreas. Transmission studies confirmed that the iridovirus is the causative agent of the disease. High mortalities occurred in 0+ juveniles in early summer and appear to be a recurrent seasonal event, whereas more sporadic outbreaks occurred in older age classes. Carrier fish were detected at low prevalence. The disease is described as epizootic haematopoietic necrosis (EHN) and is the only viral disease of fish known to occur in Australia.

Histopathology associated with pyridoxine deficiency in Atlantic salmon ( Salmo salar)

Fingerling Atlantic salmon ( Salmo solar) fed a production diet deficient in pyridoxine showed several significant effects: increased mortality; behavioral changes; degenerative changes in kidneys, ovaries, and liver; a paucity of thyroid colloid; and hyperplasia of renal hematopoietic tissue. Changes in nerve tissue were equivocal.

Histopathological Study on Vibriosis of the Salmonids

Vibrio sp. RT group infection had broken out among the salmonids in autumn 1974. Histopathological observation was made on 15 diseased rainbow trout yearlings and 5 diseased amago salmon yearlings. Diseased fish showed a small boil or erosive lesions in the body surface or/and in the head in early stages but advanced cases showed large ulcerative lesions in the body surface or/and in the head, exophthalmos and enlarged spleen.Histopathologically an early lesion was formed in the skin in which considerable bacterial multiplication, edematous dissociation and infiltration of inflammatory cells were observed in the dermal loose connective tissue. In the extended ulcerative lesions bacteria spread extensively through the lateral musculatures and made many large colonies in the interstitial tissues. In these lesions dermal necrosis associated with epithelial slough and considerable edema, slight hemorrhages, infiltration of inflammatory cells, degeneration or necrosis of muscle fibers in the affected musculature were observed. In the lesion formed in the head bacteria diffusely spread through subcutaneous connective tissue and adipose tissue and edema, slight hemorrhage, infiltration of inflammatory cells and tissues necrosis were caused. In an eye ball showing exophthalmos, which was usually associated with lesions formed in the head, marked dilatation of coroid capillaries, detachment of the retina accompanied by necrosis of the pigmented layer, iris hemorrhage and degeneration of the conea were observed. In cases of systemic infection metastatic lesions appeared in the heart and spleen, and bacterial emboli were caused in sinusoids of the liver and renal hematopoietic tissue and in lamellae capillaries.