Role of urine in the spawning of female rose bitterling,Rhodeus ocellatus ocellatus

This study clarified the spawning mechanism of female rose bitterling, Rhodeus ocellatus ocellatus, with special emphasis on the physical role of urine. Ovulated females ingested a significantly greater quantity of water by drinking than non-ovulated fish. The body weight of ovulated females increased about 1.3% while it decreased 1.0% in non-ovulated females. Urine volume in the urinary bladder increased rapidly before spawning, synchronized with the spawning cycle. On the other hand, little urine remained in specimens which had completed spawning. Histological observation demonstrated that the oviduct and the ureter joined at the proximal part of the ovipositor. On the basis of these anatomical and physiological results, it is proposed that urine plays the physical role of pushing the ovulated eggs through the elongated ovipositor during spawning.     

Recruitment,distribution boundary and habitat temperature of an arcto‐boreal gadoid in a climatically changing environment: a case study on Northeast Arctic haddock (Melanogrammus aeglefinus)

Climate change is expected to have major effects on the distribution and abundance of fish. In spite of extensive research on the topic in high‐latitude marine ecosystems, the mechanistic understanding of how temperature impacts recruitment and distribution of arcto‐boreal fish stocks remains elusive. Exemplified by an arcto‐boreal gadoid in the Barents Sea, the Northeast Arctic (NEA) haddock (Melanogrammus aeglefinus), we investigate the effect of ecosystem temperature (here temperature from a fixed reference section) on abundance and distribution boundaries between 1981 and 2008. During this time interval there has been a trend of increasing temperature in the ecosystem. We compare the ecosystem temperature with the species habitat temperature of NEA haddock (i.e., ambient temperature of the population) – two temperature approaches representing the indirect and direct environmental impacts on fish, respectively. In addition to the temperature effects, density‐dependent effects on distribution boundaries are considered. The study is based on swept area density estimates and spatial temperature data collected annually in winter surveys. We found a positive relationship between ecosystem temperature and abundance, a connection related to both direct and indirect mechanisms with short‐term and long‐term pathways. Distribution boundaries are, on a year‐to‐year basis, more related to abundance than ecosystem temperature. The long‐term trends, however, indicate a north‐eastward shift in distribution boundaries, probably indirectly related to the coinciding ecosystem temperature increase. In spite of the gradual increase in ecosystem temperature, the abundance of 4‐ to 7‐ year old NEA haddock expanded into colder waters. Thus, our results show how different the two temperature approaches may be.     

Serum lipoproteins in haddock, Melanogrammus aeglefinus L.

The lipoprotein and lipid composition of the serum in postabsorptive haddock was characterized. A low level of very low density lipoproteins (VLDL; <50 mg dL^?1) was observed in serum. High density lipoprotein (HDL) was the predominant lipoprotein class followed by low density lipoproteins (LDL). Of the lipoprotein classes, the highest proportions of triacylglycerol (28%) and phospholipid (64%) were observed in the lipid of VLDL and HDL, respectively. Serum lipid (48%), phospholipid (50%) cholesterol ester (65%) and HDL (51%) contained a high level of polyunsaturated FA, particularly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Serum triacylglycerol (38%) and VLDL (36%) contained a lower proportion of polyunsaturated fatty acid. The phospholipid, triacylglycerol, cholesterol ester and cholesterol contents of the whole serum were 739, 214, 201 and 141 mg dL^?1 respectively. On the basis of total acyl carbon number, the 38C (36%) and 36C (24%) molecules were the predominant phospholipids. The 50C (11%), 52C (17%), 54C (21%), 56C (21%) and 58C (15%) molecules were the major triacylglycerols. High molecular weight apo B‐like proteins were observed in the VLDL, LDL and intermediate density lipoprotein (IDL) fractions. An apo AI‐like protein was predominant in the HDL fraction. The prevalence of enlarged fatty livers in cultured gadoid fish (i.e. haddock and cod) is a major constraint to their commercialization and has been associated with the level of dietary lipid. Information on lipid (triacylglycerol) transport out of the liver through the serum as lipoproteins (VLDL) has implications for our understanding of fatty liver development in cultured gadoid fish. The low level of VLDL triacylglycerol (<12 mg dL^?1) circulating in the serum of postabsorptive haddock suggests either a low level of lipid transport out of the liver as triacylglycerol or relatively rapid turn over of VLDL.     

Characterization of rainbow trout milt collected with a catheter: semen parameters and cryopreservation success

Collection of fish milt by stripping risks the danger of milt contamination by urine. This may seriously influence milt characteristics and quality, including usefulness for cryopreservation. Urine contamination of milt may be avoided by using a catheter for sperm collection. The objectives of this study were to provide basic characteristics of milt collected with a catheter, to test the usefulness of this milt for cryopreservation, and to correlate characteristics of fresh and cryopreserved semen with sperm fertility rates. Milt from 25 rainbow trout Oncorhynchus mykiss (Walbaum) males were used. All samples were cryopreserved using the pellet method within 1 h of collection, using 0.6 m sucrose and 10% dimethyl sulphoxide (DMSO) as an extender. Catheterization resulted in semen of very good motility (> 90% motile spermatozoa) and high fertilization rates after cryopreservation (mean fertilization rate 81.8 ± 13.3% of control, at a sperm/egg ratio of 2.4 ± 0.3 × 10⁶). Osmolality of seminal plasma and concentrations of sodium, potassium and magnesium ions had low variability, which suggests that they are important for creating a stable environment for sperm storage in the sperm duct. Higher variability of certain seminal plasma characteristics, such as protein concentration and antiproteinase activity, suggests that these characteristics are related to individual semen features of particular males. A strong correlation of seminal plasma zinc concentration with protein concentration may reflect an importance of zinc in semen biology. Cryopreservation caused a significant release of protein and acid phosphatase from spermatozoa. Our results did not reveal any single characteristic of semen collected by catheter that could be used as a powerful predictor of cryopreservation success, presumably because all samples were of high quality.     

Renal threshold for urinary glucose excretion by tilapia in response to orally administered carbohydrates and injected glucose

Two experiments, oral carbohydrate administration (Experiment 1) and vein glucose injection (Experiment 2), were conducted to gain more insight into the ability of hybrid tilapia, Oreochromis niloticus × O. aureus, to utilize different carbohydrates and to establish the kidney threshold for urinary glucose excretion. In Experiment 1, both glucose and starch were administered orally after the tilapia were fasted for 24 h. Plasma and urine were sampled from the fish at selected time intervals from 1 to 24 h thereafter. Higher (p<0.05) plasma and urine glucose concentrations were found in fish fed on glucose than in fish fed on starch. The concentration of plasma glucose of tilapia peaked at 3 h (25.45 mM for glucose; 8.24 mM for starch) after the oral ingestion of both carbohydrates. Maximum urinary glucose concentrations (48 mM for glucose; 10 mM for starch) in fish fed glucose and starch were at 3 and 4 h post administration. In Experiment 2, five concentrations (0, 0.08, 0.12, 0.16 and 0.24 g glucose ml^–1) of glucose solution were injected into the caudal vein of the tilapia. Urine were sampled from the fish at 30-min time intervals from 0 to 6 h after the injection. Blood was sampled at 1 h after the injection. Higher urinary glucose concentrations were observed in fish injected with 0.12 g glucose ml^–1. When the urinary glucose concentrations in fish injected with the various glucose concentrations were plotted against the plasma glucose concentrations of the fish 1 h after injection, the kidney threshold for urinary glucose excretion in tilapia appeared to be about 6 mM.     

Growth,Survival, and Whole‐body Proximate and Fatty Acid Composition of Haddock,Melanogrammus aeglefinus L., Postlarvae Fed a Practical Microparticulate Weaning Diet

Further development of high‐quality feeds for hatchery‐reared haddock in the North Atlantic would benefit from a standard formulation that can be used as a reference for hatcheries and laboratory studies. A practical microparticulate diet (PMD) developed and evaluated with newly metamorphosed juvenile haddock, Melanogrammus aeglefinus L., postlarvae is proposed. Survival of fish fed PMD was just as high (88–89%; P > 0.05) as those fed a high‐quality imported feed (Biokyowa). Alternatively, fish fed PMD had higher (P < 0.05) final fork lengths (39.5 vs. 35.1 mm), wet weights (851.3 vs. 580.2 mg) and weight gains (1637.2 vs. 1115.7%). No differences (P > 0.05) in whole‐body moisture (846–857 g/kg), ash (17–18 g/kg), or protein (101 g/kg) contents were found. Lipid content of fish fed PMD (26 g/kg) was higher (P < 0.05) than those fed Biokyowa (21 g/kg) despite PMD containing 15 g/kg lower dietary lipid, suggesting higher intake and/or lipid retention. The PMD formulation proved to be a highly suitable weaning diet for haddock postlarvae based on high feed acceptance, survival, and fish growth. Given the economic and logistical difficulties associated with importing commercial weaning feeds, this easily produced practical weaning diet has good potential for use by laboratory researchers and farm managers for hatchery‐based nutrition research with haddock postlarvae.     

Apparent digestibility of common feed ingredients by juvenile haddock, Melanogrammus aeglefinus L.

The digestible energy (DE) content and the apparent digestibility coefficients (ADCs) of nutrients in common feed ingredients available in Atlantic Canada for haddock, Melanogrammus aeglefinus, feed formulations were determined. Juvenile haddock (mean weight, 93.9±2.1 g) were held in tanks equipped with fecal collection columns and fed practical fish meal‐based diets for 5 weeks. The experimental diets consisted of a reference diet and six test diets, each containing 30% test ingredient, with all diets being supplemented with chromic oxide (Cr₂O₃, 5 g kg^?1) as the inert digestion indicator. Three marine fish by‐products, herring meal (HM), shrimp meal (SM) and crab meal (CRM) and three plant protein supplements, dehulled soybean meal (SBM), canola meal (CAM) and corn gluten meal (CGM) were the test ingredients. The DE content of HM, SBM, CGM, CRM, CAM and SM were 18.3, 18.0, 17.8, 12.4, 10.9 and 8.8 MJ kg^?1 respectively. Protein ADCs were 95.9%, 92.2%, 92.3%, 82.0%, 83.0% and 73.5% respectively. Organic matter ADCs were 95.9%, 89.0%, 72.6%, 68.9%, 58.9% and 54.9% respectively. Lipid ADCs were 97.9%, 83.0%, 57.4%, 62.0%, 87.2% and 55.8% respectively. Based upon its high crude protein content and nutrient ADC and DE content, properly processed dehulled SBM was found to be a good plant protein supplement to partially replace HM in haddock feeds.     

The kidney of rainbow trout, Salmo gairdneri Richardson, in the acute phase of viral haemorrhagic septicaemia: in vivo experiments on the renal excretion of fluid, electrolytes and protein

Abstract. Kidney function was studied in unanaesthetized rainbow trout, Salmo gairdneri Richardson, with acute infection by VHS virus strain F-l causing viral haemorrhagic septicaemia. The experiments were performed in diseased and control fish with catheters chronically implanted in the dorsal aorta and the urinary bladder. Glomerular clearance of polyfructosan, tubular handling of fluid and electrolytes and urinary excretion of protein were studied. In diseased fish, plasma concentrations of inorganic solutes were reduced, but plasma osmolarity and systemic blood pressure remained unchanged. Glomerular filtration rate and urine flow, both linearly correlated, were reduced from 5.20±0.40 to 3.08±0.66 ml/h/kg and from 2.51±0.15 to 1.63±0.28 ml/h/kg, respectively (±SEM). The urine osmolarity was augmented due to increased Na and Mg concentrations in the urine. Net tubular secretion of Mg was observed in diseased fish whereas net reabsorption of Mg prevailed in the controls. Decreased renal protein excretion rate prevented a renal loss of protein and, therefore, diminished plasma protein concentrations in VHS-infected fish may be attributed to extrarenal pathophysiological mechanisms.     

Nephroblastoma in bester sturgeon,a cultured hybrid of Huso huso × Acipenser ruthenus: Diagnostic imaging,clinical and histopathological study

Abdominal distention occurred at an incidence of 1% (15 from 1500 fish) in the population of 1‐year‐old bester (Huso huso × Acipenser ruthenus). Computed tomography (CT) images and radiographs showed a soft tissue mass compressed the posterior part of the swim bladder. Ultrasonography showed that the masses had different patterns. Internal examination revealed the abdominal cavities to be filled with large masses which appeared to encompass most of the visceral organs, including the swim bladder. The masses originated from the posterior kidney. Histologically, the masses were composed of mixtures of embryonal epithelial (tubules and glomeruli), blastema and mesenchymal tissues. The tubules showed cystic, papillary and tubular patterns. Tubules and glomeruloid structures were surrounded by proliferating blastema cells. The primitive mesenchyme was composed of loose streams and whorls of spindle to stellate cells with elongate nuclei. Histological findings in the skeletal muscles, hypoderm and spleen confirmed the metastatic tumour from the kidney in two cases. Immunohistochemically, neoplastic cells of the tubules and glomeruloid structures were positive for cytokeratin AE1/AE3. Sections stained with Masson's trichrome showed blue staining of the stroma. The histopathologic findings were consistent with nephroblastoma.     

Spraguea (Microsporida: Spraguidae) infections in the nervous system of the Japanese anglerfish, Lophius litulon (Jordan), with comments on transmission routes and host pathology

Anglerfish from the genus Lophius are a globally important commercial fishery. The microsporidian Spraguea infects the nervous system of these fish resulting in the formation of large, visible parasitic xenomas. Lophius litulon from Japan were investigated to evaluate the intensity and distribution of Spraguea xenomas throughout the nervous system and to assess pathogenicity to the host and possible transmission routes of the parasite. Spraguea infections in L. litulon had a high prevalence; all fish over 403 mm in standard length being infected, with larger fish usually more heavily infected than smaller fish. Seventy percent of all fish examined had some gross visible sign of infection. The initial site of development is the supramedullary cells on the dorsal surface of the medulla oblongata, where all infected fish have parasitic xenomas. As the disease progresses, a number of secondary sites typically become infected such as the spinal, trigeminal and vagus nerves. Fish with infection in the vagus nerve bundles often have simultaneous sites of infection, in particular the spinal nerves and along the ventral nerve towards the urinary bladder. Advanced vagus nerve infections sometimes form xenomas adjacent to kidney tissue. Spraguea DNA was amplified from the contents of the urinary bladders of two fish, suggesting that microsporidian spores may be excreted in the urine. We conclude that supramedullary cells on the hindbrain are the primary site of infection, which is probably initiated at the cutaneous mucous glands where supramedullary cells are known to extend their peripheral axons. The prevalence of Spraguea infections in L. litulon was very high, and infections often extremely heavy; however, no associated pathogenicity was observed, and heavily infected fish were otherwise normal.     

Interannual differences in larval haddock survival: hypothesis testing with a 3D biophysical model of Georges Bank

The ultimate goal of early life studies of fish over the past century has been to better understand recruitment variability. As evident in the Georges Bank haddock (Melanogrammus aeglefinus) population, there is a strong relationship between recruitment success and processes occurring during the planktonic larval stage. This research sought new insights into the mechanisms controlling the recruitment process in fish populations using biological–physical modeling methods together with laboratory and field data sets. We created the first three‐dimensional model of larval haddock on Georges Bank by coupling models of hydrodynamics, lower trophic levels, a single copepod species, and larval haddock. Interactions between feeding, metabolism, growth, vertical behavior, advection, predation, and the physical environment of larval haddock were quantitatively investigated using the coupled models. Particularly, the model was used to compare survival over the larval period and the sources of mortality in 1995 and 1998, 2 years of disparate haddock recruitment. The results of model simulations suggest that the increased egg hatching rates and higher food availability, which reduced starvation and predation, in 1998 contributed to its larger year‐class. Additionally, the inclusion of temperature‐dependent predation rates produced model results that better agreed with observations of the mean hatch date of survivors. The results from this biophysical model imply that food limitation and its related losses to starvation and predation, especially from hatch to 7 mm, may be responsible for interannual variability in recruitment and larval survival outside of the years studied.     

四指马泌尿系统胚后发育组织学研究

为了解四指马鲅(Eleutheronema tetradactylum)泌尿系统胚后发育的一般规律,采用石蜡组织连续切片和H-E染色技术,对1~50 dph(days post hatching)的四指马鲅泌尿系统胚后发育进行了研究。结果显示,四指马鲅前肾小管在3 dph已经形成,中间分布少量未分化干细胞;至7 dph前肾小管刷状缘明显,泌尿机能增强;15 dph前肾小管继续分化,管壁细胞结构清晰;20 dph观察到少量前肾小管退化,泌尿机能开始衰退;前肾在发育过程中未观察到肾小体形成。中肾肾小管原基在7 dph开始形成;20~25 dph中肾早期肾小体和肾小管芽形成;30 dph肾小管数量剧增,亚成熟肾小体形成,近端小管和远端小管开始区分;45 dph第一近端小管和第二近端小管开始区分,至50 dph观察到成熟肾小体;中肾有较多黑色素-巨噬细胞中心、淋巴细胞和红细胞分布,兼具造血、免疫及泌尿功能。中肾管为输尿管,始于中肾后端,并行向后合并膨大为输尿管膀胱。     

An 11-ketotestosterone induced kidney-secreted protein: the nest building glue from male three-spined stickleback, Gasterosteus aculeatus

In male three-spined sticklebacks, Gasterosteus aculeatus, the kidney hypertrophies during the breeding season and produces a glue which is used in nest-building. This hypertrophy is androgen dependent with 11-ketotestosterone (11 KT) being most effective. The aim of the present study was to characterize the protein composition of this glue. Threads of glue were collected from stickleback nests and glue material was sampled from the content of urinary bladders of male sticklebacks in breeding condition. The samples were investigated using sodium dodecyl sulphate- polyacrylamide gel electrophoresis (SDS-PAGE). One major glycoprotein dominated in both the nest-threads and urinary bladder samples. The identified glycoprotein had a molecular mass of approximately 203 kDa. After deglycosylation the molecular mass was approximately 200 kDa. The amino acid composition of the protein from urinary bladder content was almost identical to the amino acid composition of the protein from the nest-threads. The protein had a relatively high content of cysteine (7.6–8.0%). The glycoprotein was named spiggin. Spiggin was absent in the urinary bladder of untreated castrated fish, but spiggin was present in sham- operated fish and in castrated fish treated with 11 KT. These results demonstrate that spiggin is induced by 11 KT. Spiggin is so far the only protein known to be induced by 11KT and based on the present findings we suggest that spiggin represents a novel structural protein.     

Urinary bladder displacement (lateroflexio vesicae urinariae) in the sow

In older gravid sows in the last stage of gestation or after pariturition lateroflexion of the urinary bladder into the space between the vagina and the pelvic wall occasionally occurs. The reposition of the displaced urinary bladder by exerting external pressure either on the vulva or the perineum is usually unsuccessful. Only after the bladder has been emptied by a catheter spontaneous reposition does occur. In two cases displacement of the urinary bladder occurred immediately after normal pariturition. In both cases relapses were observed after the removal of the implanted balloon catheter. Both sows had the urinary balloon catheter reinserted and it was left in place until the weaning of the piglets. There were no complications during the whole lactation period. Both sows reared their piglets and could then be slaughtered.     

Assessment of sex and evaluation of ovarian maturation of fish using ultrasonography

Ultrasonography is a non‐invasive method that can be used to assess sex and maturity of fish. This study was conducted to examine the feasibility of using ultrasound images to sex juvenile and mature halibut Hippoglossus hippoglossus (Linnaeus), mature winter Pleuronectes americanus (Walbaum) and yellowtail flounder Pleuronectes ferruginea (Storer) and mature haddock Melanogrammus aeglefinus (Linnaeus). In females, both immature and mature ovaries were relatively easy to distinguish, but in males only mature testes were consistently distinguishable. Ovarian maturation of haddock and ovulatory cycles of halibut may also be assessed using ultrasound.     

The growth of larval cod and haddock in the Irish Sea: a model with temperature,prey size and turbulence forcing

We applied a physiological individual‐based model for the foraging and growth of cod (Gadus morhua) and haddock (Melanogrammus aeglefinus) larvae, using observed temperature and prey fields data from the Irish Sea, collected during the 2006 spawning season. We used the model to estimate larval growth and survival and explore the different productivities of the cod and haddock stocks encountered in the Irish Sea. The larvae of both species showed similar responses to changes in environmental conditions (temperature, wind, prey availability, daylight hours) and better survival was predicted in the western Irish Sea, covering the spawning ground for haddock and about half of that for cod. Larval growth was predicted to be mostly prey‐limited, but exploration of stock recruitment data suggests that other factors are important to ensure successful recruitment. We suggest that the presence of a cyclonic gyre in the western Irish Sea, influencing the retention and/or dispersal of larvae from their spawning grounds, and the increasing abundance of clupeids adding predatory pressure on the eggs and larvae; both may play a key role. These two processes deserve more attention if we want to understand the mechanisms behind the recruitment of cod and haddock in the Irish Sea. For the ecosystem‐based management approach, there is a need to achieve a greater understanding of the interactions between species on the scale a fish stock is managed, and to work toward integrated fisheries management in particular when considering the effects of advection from spawning grounds and prey–predator reversal on the recovery of depleted stocks.     

Selectivity and escape percentages during three phases of the towing process for codends fitted with different selection systems

We investigated the selectivity of cod (Gadus morhua) and haddock (Melanogrammus aeglefinus) escaping from three different selection systems (a conventional diamond-mesh codend, a codend fitted with escape windows, and a codend fitted with a sorting grid) using the covered codend technique. By using a MultiSampler attached to the codend cover to partition the collection of escaping fish, we were able to determine the partitioned selectivity of each of these devices at different phases during the towing process: at depth, during the haul back, and at the surface.The percentages of escaping fish at depth from the diamond-mesh codend were significantly lower than those estimated for the escape windows and sorting grid codends: almost 62% of cod and 79% of haddock escaping from the diamond-mesh codend did so at depth, whereas approximately 75% of cod and 93% of haddock escaped from the other two systems at depth. The partitioned selectivity parameters, l₅₀ and SR, were estimated and modelled for each of the three phases of the towing process and used to compare the selective properties of the three selection systems. A comparison was also made with a traditional whole haul selectivity analysis. Finally, bulk catch was added to the model as an explanatory variable and its effect upon selectivity was measured in both, partitioned and whole haul estimates.     

Reabsorption of urea by the kidney of the freshwater rainbow trout

In light of recent evidence that carrier-mediated transport of urea occurs in the mammalian kidney, this study examined the renal handling of urea in freshwater rainbow trout (Oncorhynchus mykiss). Fish were fitted with indwelling arterial and urinary bladder catheters for the measurement of plasma and urine composition (urea, Na⁺, Cl^-, glucose, H₂O), glomerular filtration rate ([³H]PEG-4000), and urine flow rate, thereby allowing quantification of tubular reabsorption rates. The fractional reabsorption of urea (72%) was greater than that of H₂O (50%) but less than that of Na⁺, Cl^-, or glucose (95–100%) and occurred against an apparent concentration gradient, suggesting active reabsorptive transport; [urea] in the urine was only 59% of that in blood plasma. When fish were infused with exogenous urea loads, these patterns remained largely unchanged, and urea reabsorption increased in direct proportion to the filtered urea load. There was no evidence for saturation of the reabsorptive transport mechanism at urea filtration rates up to 4-fold above the normal range, representing urea loading rates that proved toxic. Extra-renal excretion, presumably through the gills, increased markedly, almost keeping pace with urea loading. This evidence suggests that carrier-mediated reabsorptive transport of urea occurs in the kidney and that plasma urea levels are normally subject to tight homeostatic control in freshwater trout.     

Multi‐decadal variations in spawning ground use in Northeast Arctic haddock (Melanogrammus aeglefinus)

Despite the importance both in an ecological and management context, much uncertainty remains about the underlying factors controlling spawning ground use in marine fish. Here, we investigate how spawning ground use of Northeast Arctic (NEA) haddock (Melanogrammus aeglefinus) has changed over time. By combining data from a Soviet‐Russian egg survey conducted over a 35 year period (1959–1993) with a biophysical drift model, we estimate annual median spawning latitudes of NEA haddock. Furthermore, we test for statistical association between the median latitude of spawning and covariates that are candidates for shaping the spawning distribution of haddock. The results indicate no association with temperature, as measured at the Kola transect, or demographic structure, as measured by the mean weight of spawners. However, we find a positive and significant association between spawning latitude and spawning stock biomass, indicating that density‐dependence may be more important in shaping spawning location than other factors such as temperature and demographic structure. A potential mechanism for the reported effect of spawning stock biomass may act through the distribution at the feeding grounds, which is reported to expand north‐ and eastwards at high population density.     

Tissue localization of Aeromonas salmonicida in Atlantic salmon, Salmo salar L., following experimental challenge

Three groups of Atlantic salmon, Salmo salar L., were exposed to live, colony-forming, radiolabelled Aeromonas salmonicida bacteria in a bath challenge: (1) fish with artificial wounds; (2) fish with a reduced epidermal mucus layer caused by removal of the mucus layer on two occasions by a swabbing procedure; and (3) a control group of untreated fish. Fish were killed 2, 6 and 24 h after challenge, and radioactivity (cpm g^–1) was measured in the blood, mucus, skin, wound area, gills, anterior kidney, posterior kidney, spleen, midgut and hindgut. The highest levels of radioactivity were measured in the wound areas and in the gills. There was a significant positive correlation between the levels of radioactivity in the gills and blood, and between the mucus and skin at 2 h post-challenge. Two hours after the bath challenge, live A. salmonicida bacteria were found in the blood of fish in the 'swabbed' and 'artificial wound' groups, and not in the control group. Twenty-four hours after the bath challenge, the kidney of fish from all groups contained viable bacteria, whereas the blood was negative.