Diagnostic criteria for proliferative hepatic lesions in brown bullhead Ameiurus nebulosus

Brown bullhead Ameiurus nebulosus is used as indicator species for contaminant effects at areas of concern (AOC) in the Great Lakes and other areas. One of the beneficial use impairments at numerous AOC is 'fish tumors and other deformities'. An impairment occurs when the prevalence of fish tumors and other deformities exceeds those at unimpacted or control sites or when survey data confirm the presence of neoplastic or preneoplastic liver lesions in bullhead or white sucker Catostomus commersonii. Numerous surveys have been conducted over the years assessing neoplasia in these fishes, both liver and skin tumors. However, a major problem in comparing the results has been a lack of consistent criteria for evaluating histological changes in bullhead livers. As individual AOC develop and implement remedial action plans, realistic and attainable delisting targets need to be specified. For this to occur and be consistent from site to site there must be standardization of the criteria being used to evaluate specific impairments. In this report, specific diagnostic criteria are provided for both non-neoplastic and neoplastic proliferative hepatocellular and biliary lesions. These criteria should assist fish pathologists in describing and categorizing proliferative liver lesions from brown bullhead.     

An investigation of carbonic anhydrase activity in the gills and blood plasma of brown bullhead (Ameiurus nebulosus), longnose skate (Raja rhina), and spotted ratfish (Hydrolagus colliei)

Separated plasma and whole blood non-bicarbonate buffering capacities, together with plasma and gill carbonic anhydrase activities and endogenous plasma carbonic anhydrase inhibitor activity were investigated in three species of fish: the brown bullhead (Ameirus nebulosus), a teleost; the longnose skate (Raja rhina), an elasmobranch; and the spotted ratfish (Hydrolagus colliei), a chimaeran. The objective was to test the hypothesis that species possessing gill membrane-bound carbonic anhydrase and/or plasma carbonic anhydrase activity would also exhibit high plasma nonbicarbonate buffering capacity relative to whole blood non-bicarbonate buffering capacity and would lack an endogenous plasma carbonic anhydrase inhibitor. Separated plasma non-bicarbonate buffering capacity constituted > or = 40% of whole-blood buffering in all three species. In addition, all species lacked an endogenous plasma carbonic anhydrase inhibitor. Separated plasma from skate and ratfish contained carbonic anhydrase activity, whereas bullhead plasma did not. Examination of the subcellular distribution and characteristics of branchial carbonic anhydrase activity revealed that the majority of branchial carbonic anhydrase activity originated from the cytoplasmic fraction in all species, with only 3-5% being associated with a microsomal fraction. The microsomal carbonic anhydrase activity of bullhead and ratfish was significantly reduced by washing, indicating the presence of carbonic anhydrase activity that was not integrally associated with the membrane pellet, microsomal carbonic anhydrase activity in skate was unaffected by washing. In addition, microsomal carbonic anhydrase activity from skate and ratfish but not bullhead gills was released to a significant extent from its membrane association by treatment with phosphatidylinositol-specific phospholipase C. The results obtained for skate are consistent with published data for dogfish, suggesting that the possession of branchial membrane-bound carbonic anhydrase activity may be a generalised elasmobranch characteristic. Ratfish, which also belong to the class Chondrichthyes, exhibited a similar pattern. Unlike skate and ratfish, bullhead exhibited high plasma non-bicarbonate buffering capacity and lacked an endogenous carbonic anhydrase inhibitor in the absence of plasma and gill membrane-bound carbonic anhydrase activities.     

Apparent diffusion limitations on branchial CO2 transfer are revealed by severe experimental anaemia in brown bullhead (Ameiurus nebulosus)

In this study, the rapid (within 2 h) effects of acute anaemia on blood gas and acid-base status, as well as cardiorespiratory variables, were examined in brown bullhead (Ameiurus nebulosus). Anaemia was induced by blood withdrawal coupled to volume replacement with saline. Lowering haematocrit from the control value of 23.5+/-1.0% (mean+/-S.E.M.; N=37) to 5.9+/-0.3% (N=37) resulted in a significant increase (by 2.63+/-0.51 torr; N=7) in arterial CO(2) tension (PaCO(2)) over the subsequent 2-h period in the absence of a change in arterial O(2) tension (PaO(2)). Treatment with bovine carbonic anhydrase (CA) reduced the extent of the PaCO(2) increase to the point where it was not statistically significant. In both control and CA-treated fish, arterial pH decreased during acute anaemia; the acidosis was of mixed respiratory and metabolic origin in control fish and primarily metabolic in CA-treated fish. Inducing anaemia caused increases in both cardiac output (V*b) and heart rate that were similar in control and CA-treated fish. Experimental elevation of V*b equivalent to that observed during anaemia, but in the absence of lowered haematocrit, increased PaCO(2) significantly by 1.49+/-0.74 to 1.64+/-0.78 torr (N=5) without affecting PaO(2). These findings suggest that CO(2) excretion in bullhead, as in rainbow trout, is effectively diffusion-limited, and that approximately half of the increase in PaCO(2) measured during the initial 2 h of anaemia results from the impact of increased blood flow (hence decreased gill transit time) in a diffusion-limited system.     

Fine structure of the axillary gland in the brown bullhead (Ictalurus nebulosus)

The axillary glands of Ictalurus are lobulated invaginations of the epidermis, opening at a pore between the pectoral spine and the cleithrum. Holocrine cells lining a false lumen form a viscous secretion. The secretory cells originate in the tenuous basal layer of the gland wall. Secretion is initiated by the formation of compound vesicles in cells that become very large and have complex cytoplasm of a varied appearance. Golgi systems are well developed and the perinuclear cytoplasm may contain many mitochondria and sacs of ribosomal endoplasmic reticulum; some tracts of cytoplasm are vesicular and contain free ribosomes. Some cells contain numerous large lysosomes, and some have extensive contents of fibrillar masses imperfectly separated by membranes, that recall the appearance of the mucous secretion of goblet cells. The secretory cells break down, releasing the degenerating organelles, including the nuclei, into the false lumen. Some structures are still recognizable in the secretion even after it has been expelled, but the main part of the formed secretion consists of the mucus-like masses. Various leucocytes are found in the gland walls and embedded in the secretion. The fine structure differentiates the holocrine cells of the axillary gland from the club cells of the epidermis, and from the venom glands associated with the fin spines of catfishes. The function of the axillary gland secretion remains unknown.     

The significance of biparental care in the brown bullhead,Ictalurus nebulosus

Synopsis Parental care in the brown bullhead is characterized by variation in the participation of each sex. Most broods are attended by both sexes, but some are attended by a male alone, or rarely, a female alone. Two care-givers were more successful than one alone in fostering offspring survival. However, there was no significant difference between two care-givers and one alone in the proportion of time that broods were unattended. Potential brood predators were chased less frequently by one adult alone than by adults aided by their mate. This difference may be unimportant since two adults simultaneously attended their brood only 19% of the time. Males alone attended their broods a significantly greater proportion of time than did either males or females aided by their mates. This difference suggests that males alone sustain a greater cost of care-giving (starvation and therefore reduced future reproduction) than do males aided by their mates. Thus, males alone may more often leave broods (and not return) than males that are aided in care-giving. The differential success observed may be due to a difference in the likelihood that the male (the principal care-giver) leaves the brood permanently, rather than differences in the quality of care one or two adults provide. I suggest that two care-givers are more successful than one because the net benefits of care-giving exceed the net benefits of leaving for males when aided by their mates.     

Evidence for a morphological component in acid-base regulation during environmental hypercapnia in the brown bullhead (Ictalurus nebulosus)

Summary Exposure of adult brown bullheads Ictalurus nebulosus (120–450 g) to environmental hypercapnia (2% carbon dioxide in air) and subsequent recovery caused transient changes in whole body net sodium flux (J _net ^Na+ ) and net chloride flux (J _net ^Cl- ) resulting largely from changes in whole body sodium influx (J _in ^Na+ ) and chloride influx (J _in ^Cl- ). Scanning electron microscopy (SEM) revealed that the fractional area of chloride cells (CCs) on the interlamellar regions was reduced by 95% during environmental hypercapnia. During post-hypercapnic recovery, gill filament CC fractional area increased. The changes in J _in ^Cl- during and after environmental hypercapnia were closely associated with the changes in CC fractional area while the changes in J _in ^Na+ did not correspond to the changes in CC fractional area. Transmission electron microscopy (TEM) supported the SEM observations of CC surface area changes and demonstrated that these changes were caused by covering/uncovering by adjacent pavement cells (PVCs). Lamellar and filament PVC microvilli density increased during hypercapnia while there was a subsequent reduction in the post-hypercapnic period. These data suggest that an important mechanism of acid-base regulation during hypercapnic acidosis is modification of the chloride cell-associated Cl^-/HCO ₃ ^- exchange mechanism. We suggest that bullheads vary availability, and thus functional activity, of this transporter via reversible morphological alterations of the gill epithelium. The increase in density of PVC microvilli may be associated with sodium uptake and/or acidic equivalent excretion during acidosis.     

Incidence of antibiotic and metal resistance and plasmid Carriage in Aeromonas isolated from brown bullhead (latalurus nebulosus)

G.W. PETTIBONE, J.P. MEAR AND B.M. SAMPSELL. 1996. Seventy-four strains of Aeromonas were isolated from skin, intestine, kidney and liver of 16 brown bullhead ( Ictalurus nebulosus ). All strains demonstrated multiple antibiotic resistance (MAR) with most strains resistant to rifampin (97%), novobiocin (96%) and vancomycin (85%). The minimum inhibitory concentration of eight metals to selected strains of Aeromonas revealed an apparent toxicity of chromium > copper > cadmium > nickel > mercury > zinc > cobalt > lead, based on the percentage of isolates sensitive to each concentration of metal. Plasmid DNA was found in 36% (27) of the isolates and most plasmid-containing strains had multiple plasmids less than 12 kilobase pairs (kbp) in size. No relationship between plasmid content and antibiotic or metal resistance was found. Plasmid incidence in bacteria isolated from five fish indicated that plasmids were more likely to occur in strains isolated from kidney and liver than in strains isolated from skin and intestine.     

Impact of black bullhead (Ameiurus melas) on turbidity in a diked wetland

Benthivorous fish in shallow, aquatic systems have been correlated with increased turbidity and declines in macrophyte production and wildlife use. Bullheads have been credited with increasing turbidity, but this has been seldom tested and has not been studied in a diked marsh. To assess the relationships of black bullhead (Ameiurus melas) and turbidity, we assembled mesocosmsin the Show Pool Management Unit of The Ottawa National Wildlife Refuge, OH, U.S.A. We stocked treatment enclosures with different biomasses of black bullhead at weekly intervals. Mean turbidity within treatment enclosures was significantly higher than within controls but remained lower than that of the open marsh. Both surface and bottom turbidity increased with adult and juvenile black bullhead biomass. Turbidity increased with fine sand concentration only in the presence of juvenile fish. Wind speed and direction were significant influences on the turbidity of the open marsh, but not within control enclosures. That treatment turbidity – even at extreme biomasses – remained significantly lower than the turbidity of the open marsh implicates fetch in having a greater influence on a marsh's turbidity than the presence of black bullhead. The greater impact of benthivorous fish on turbidity within shallow systems may be an indirect one through the destruction of macrophytes and subsequent destabilization of unconsolidated substrates.     

Metabolism of benzo[a]pyrene and persistence of DNA adducts in the brown bullhead (Ictalurus nebulosus).

1. The in vitro metabolism of [3H]benzo[a]pyrene (BP) and [14C]benzo[a]pyrene-7,8-dihydrodiol (BP-7,8-diol) by liver of brown bullhead (Ictalurus nebulosus) was characterized, as was the formation and persistence of BP-DNA adducts in vivo. 2. Compared to rat liver microsomes, bullhead liver microsomes produced relatively larger amounts of BP-7,8-diol (predominantly the [-] enantiomer) and smaller amounts of of BP-7,8-diol (predominantly the [-] enantiomer) and smaller amounts of BP-4,5-diol. 3. BP phase I metabolites were efficiently converted by freshly isolated bullhead hepatocytes to conjugates, predominantly glucuronides. 4. BP-7,8-diol was metabolized by hepatocytes 4-fold more rapidly than was BP and was converted to approximately equal amounts of glucuronides, glutathione conjugates and sulfates. 5. BP-DNA adducts formed in bullhead liver with a lag time of several days and maximum adduct formation at 25-30 days. The major adduct was anti-BPDE-deoxyguanosine.     

Vertical distribution and movements of brown bullhead (Ameiurus nebulosusLesueur 1819) in Motuoapa Bay,southern Lake Taupo,New Zealand

Ultrasonic telemetry was used to determine the swimming depths of brown bullhead (Ameiurus nebulosus) in Motuoapa Bay at the southern end of Lake Taupo, New Zealand. Sonic transmitters equipped with a pressure sensor were surgically implanted in 12 adult fish (295–345 mm FL). Three automatic receivers recorded a total of 61335 data points on swimming depth between 26 November 1998 and 7 November 1999. Brown bullheads swam in water from 0 to 17 m deep but showed some seasonal variations. They were more active at night when they used shallow water extensively, especially during the winter months. They were most mobile in spring when they left Motuoapa Bay, suggesting migratory behaviour. During winter the fish remained more stationary, only moving around inside the bay. The results of this study provide guidance for establishing a brown bullhead control program should it become necessary.     

Seasonal changes in serum thyroid hormone levels of the feral brown bullhead, Ictalurus nebulosus Lesueur

Seasonal changes in serum L-thyroxine (T4) and triiodo-L-thyronine (T3) concentrations were measured in a feral population of brown bullheads, Ictalurus nebulosus . Serum T4 was elevated in mid April concomitant with the onset of gonadal recrudescence and the increase in ambient temperatures from the winter level. Thereafter, serum T4 levels were constant throughout the late spring and summer (except for a significant lowering in levels associated with the spawning and early postspawning period), declining to a low overwintering level by mid December. Serum T3 levels increased in April, and apart from a significant decrease in July associated with the postspawning period, rose progressively until August, after which levels declined progressively to reach overwintering levels again by mid December. Between mid April and mid May serum T4/T3 ratios fell from the high values found in fish during the winter months. Apart from variations associated with the fluctuations of T4 and T3 concentrations in the spawning and early postspawning periods, serum T4/T3 ratios remained at a low level throughout the summer and early autumn months, and rose again in October to the overwintering levels. The circannual cycles are discussed in relation to the possible role of the thyroid hormone in gonadal maturation and metabolism in this species.     

Gill morphology during hypercapnia in brown bullhead (Ictalurus nebulosus): role of chloride cells and pavement cells in acid-base regulation

The role of gill chloride cells (CCs) and pavement cells (PVCs) in acid-base regulation was evaluated in brown bullhead catfish (Ictalurus nebulosus) subjected to acute hypercapnia (water Pco₂=15 torr). Chronic (10 day) cortisol treatment was used as a tool to cause CC proliferation to permit a comparison of the regulatory capacities in groups of fish with widely different gill CC populations. Cortisol (4mg kg^?1 day^?1) caused a pronounced increase (170%) in the surface area of CCs exposed to the water based on scanning and transmission electron microscope analysis. The density of PVC apical membrane microvilli was significantly increased (20%) by cortisol treatment. Exposure of either group of fish to hypercapnia caused similar changes in gill epithelial morphology including: (i) a marked reduction in the surface area of exposed CCs (52 and 78% reduction in the control and cortisol-treated fish, respectively); and (ii) pronounced increases in PVC apical membrane microvilli density (21 and 27% in the control and cortisol-treated fish, respectively). The rates of Cl^? uptake (J_incl^?) and Na⁺ uptake (J_inNa⁺) were elevated (150 and 262%, respectively) in the cortisol-treated fish. Regardless of treatment, J_incl^? was markedly reduced to approximately the same levels after 6 h of hypercapnia, J_inNa⁺ was stimulated in the control group and reduced in the cortisol-treated group and thus, after 6 h of hypercapnia, J_inNa⁺ was equal in each group. The similar morphological responses in fish possessing different initial populations suggests that the predominant mechanism of acid-base regulation during hypercapnia, reduction of C1^?/HCO₃^? exchange, is accomplished by removal of the CC-associated C1-/HCO₃^? exchange sites from the water. The increase in PVC microvilli density during hypercapnia suggests a role for the PVC in acid-base regulation.     

Use of shape analysis in the investigation of disputable meristic characters for Ameiurus melas (Rafinesque, 1820) and Ameiurus nebulosus (Lesueur, 1819)

The aim of this paper was to use geometric morphometry as a helpful tool to clarify the overlap between meristic characters (such as the number of anal fin rays) of different fish identification keys employed for the two sister species, Ameiurus melas and Ameiurus nebulosus. Geometric morphometry included shape analysis of 15 landmarks on 432 specimens. The analyses showed significant differences among groups based on the sampling site, while there was no difference among groups based on the number of anal fin rays (used to distinguish species). This suggests that the use of meristic characters for the determination of these two sister species is rather questionable.     

Variable growth and longevity of yellow bullhead (Ameiurus natalis) in the Everglades of south Florida,USA

Yellow bullhead (Ictaluridae: Ameiurus natalis) is the most abundant ictalurid catfish in the Everglades of southern Florida, USA, and, as both prey and predator, is one of many essential components in the ecological‐simulation models used in assessing restoration success in the Everglades. Little is known of its biology and life history in this southernmost portion of its native range; the present study provides the first estimates of age and growth from the Everglades. In total, 144 yellow bullheads of 97–312 mm total length (TL) were collected from canals and marshes of the Everglades between April 2000 and January 2001, and from October 2003 to February 2005. Fish were aged using cross‐sections of pectoral spines and ranged from 1–12 years, with the maximum age almost twice that of any yellow bullhead previously reported. Yellow bullheads from south Florida grew relatively rapidly during their first 3 years, but after age 5 growth slowed and fish approached an asymptote of ～214 mm TL. Compared to other populations in the United States, yellow bullhead in the Everglades grew relatively slowly, were smaller at age overall, but survived to older ages.     

Gonadal morphology, enzyme histochemistry and plasma steroid levels during the annual reproductive cycle of male and female brown bullhead catfish, Ictalurus nebulosus Lesueur

The annual reproductive cycle of the brown bullhead catfish, Ictalurus nebulosus Lesueur, was investigated over a two-year period. In females, GSI increased in the spring as follicles enlarged and the granulosa became hypertrophied, dropped during spawning in August, then rose in the autumn as follicles enlarged slightly. 3β-Hydroxysteroid dehydrogenase (3β-HSD) activity was limited to thecal nests of large, vitellogenic follicles. Plasma testosterone and estradiol-17β levels increased in parallel with GSI. Levels of both steroids dropped prior to the spawning period, although a peak in estradiol-17β was evident during the spawning period. No 11-ketotestosterone was detected in female plasma. In males, GSI increased in the spring as spermatogenesis proceeded, and dropped during spawning. 3β-HSD activity was confined to Leydig cells and was most intense prior to spawning. Plasma testosterone and 11-ketotestosterone peaked during the pre-spawning period, dropped prior to spawning, then rose slowly during the autumn. A peak in estradiol-17β occurred during the spawning period. Significant differences in GSI and plasma steroid levels during the pre-spawning and spawning periods were observed between the two yearly cycles; they may be related to differences in rainfall during these periods.     

Seasonal changes in territoriality and frequency of agonistic behavior in two densities of juvenile brown bullhead,Ictalurus nebulosus

Synopsis Incidence of territoriality and frequency of six agonistic action patterns of juvenile brown bullhead, Ictalurus nebulosus, were observed in the laboratory over a two year period at two fish densities under conditions of constant light and temperature. High density fish showed little territoriality or agonistic behavior while low density fish displayed an annual cycle of agonistic behavior and territoriality. Agonistic behavior frequency and incidence of territoriality peaked in February and March and were higher the second year when compared with the first.     

Morphology and 18S rDNA of Henneguya gurlei (Myxosporea) from Ameiurus nebulosus (Siluriformes) in North Carolina

Henneguya gurlei was isolated from Ameiurus nebulosus captured in North Carolina and redescribed using critical morphological features and 18S small-subunit ribosomal RNA (SSU rDNA) gene sequence. Plasmodia are white, spherical, or subspherical, occur in clusters, measure up to 1.8 mm in length, and are located on the dorsal, pectoral, and anal fins. Histologically, plasmodia are located in the dermis and subdermally, and the larger cysts disrupt the melanocyte pigment layer. The spore body is lanceolate, 18.2 +/- 0.3 microm (range 15.7-20.3) in length, and 5.4 +/- 0.1 microm (range 3.8-6.1) in width in valvular view. The caudal appendages are 41.1 +/- 1.1 microm (range 34.0-49.7) in length. Polar capsules are pyriform and of unequal size. The longer polar capsule measures 6.2 +/- 0.1 microm (range 5.48-7.06), while the shorter is 5.7 +/- 0.1 microm (range 4.8-6.4) in length. Polar capsule width is 1.2 +/- 0.03 microm (range 1.0-1.54). The total length of the spore is 60.9 +/- 1.2 microm (range 48.7-68.5). Morphologically, this species is similar to other species of Henneguya that are known to infect ictalurids. Based on SSU rDNA sequences, this species is most closely related to H. exilis and H. ictaluri, which infect Ictalurus punctatus.