The immune response of the brown trout, Salmo trutta, L. to MS2 bacteriophage: immunogen concentration and adjuvants

The humoral antibody response to single intraperitoneal inoculations of MS2 bacteriophage was followed in Salmo trutta using a 50% bacteriophage neutralisation litre ( sd ₅₀) method. Immune memory was observed with enhancement of both the time and level of antibody production, though an initial suppression was observed 7 days after secondary inoculation. Antibody titres were related to MS2 concentration, and adjuvants were found to enhance the response. An increased enhancement of antibody production by Freund's complete adjuvant over that of incomplete adjuvant was observed.     

The influence of levamisole on the growth of carp (Cyprinus carpio L.) larvae

The present results indicate that levamisole stimulates the growth of carp larvae without effect on its survival and rate of development. The differences between the growth of larvae exposed to 2 mg · dm^-3 and larvae from control group were significant concerning the length and the weight of larvae.

Zusammenfassung

Der Einfluß von Levamisol anf das Wacbstum von Karpfen
Levamisol stimuliert den Zuwachs frisch geschlüpfter Karpfenbrut. Es hat keinen Einfluß auf Überle-bens-und Entwicklungsrate. Die Unterschiede im Zuwachs von Brut, die mit einer Levamisollösung (Konzentration 2 mg · dm^-3) behandelt wurden, und Brut der Kontrollgruppe waren statistisch signifikant sowohl hinsicbtlich des Gewichtes als auch der Brutlänge.

Résumé

Influence de lévamisole sur la croissance d'alevins de carpe (Cyprinus carpio)
Les résukats d'expériences faites en laboratoire montrent que lévamisole stimule la croissance d'alevins de carpe sans inluencer leur survie ou leur taux de développement. Les différences entre la croissance d'alevins soumis à une concentration de 2 mg · dm^-3, et celle du groupe contrôle étaient significatives quant à la longueur et au poids des alevins.     

The effect of dissolved cadmium on the chloride cells of the gills of the stickleback, Gasterosteus aculeatus L.

The numbers of chloride cells on the gills of three-spined sticklebacks, Gasterosteus aculeatus , have been measured after exposure of the fish to 0.5, 1.0 and 1.5 mg Cd²⁺ dm⁻³ soft water (132 mg dm⁻³ as CaCO₃) and 2, 4 and 6 mg Cd²⁺ dm⁻³ hard water (299 mg dm⁻³). The numbers of chloride cells increased with time but later tended to decline. The significance of this finding is discussed in relation to the functions of chloride cells, one of which is presumed to be to eject absorbed heavy metals such as cadmium.     

Embryonic and larval development of brown trout, Salmo trutta L.: exposure to trace metal mixtures in soft water

Freshly fertilized ova of brown trout, Salmo trutta L., were exposed to all possible mixtures of Al (6000 nmol 1¹), Cu (80 nmol 1⁻¹), Pb (50 nmol 1⁻¹) and Zn (300 nmol 1 ¹). In a separate experiment, newly hatched brown trout yolk-sac fry were exposed to Mn (1500 nmol 1⁻¹), Fe (2500 nmol 1 ¹), Ni (200 nmol 1⁻¹) or Cd (4 nmol 1 ¹), separately, and in mixtures with either Al or Cu. Both experiments were conducted in flowing, artificial softwater media nominally at pH 5.6 [Ca] 20 μmol 1 ¹ and 10° C.
Mortalities were high in fry subjected to treatments which contained both Al and Cu (31–72%), and to the Cu + Fe treatment (78%) compared with those from the other trace metal mixtures (0–22%). In all the treatments tested, fry exposed to trace metal mixtures containing Al and/or Cu had reduced whole body Ca, Na and K content, and seriously impaired skeletal calcification. Whole body Mg content was variable. In trace metal mixtures which contained Cu but not Al, the effects on fry survival and whole body mineral content were in general more deleterious than the corresponding mixtures but with Al present rather than Cu. The presence of Pb and/or Zn in mixtures with Al and/or Cu had a slight ameliorative effect in terms both of fry survival and whole body mineral content.     

Acute toxicology to an estuarine teleost of mixtures of cadmium, copper and zinc salts

Effects of mixtures of chloride salts of cadmium, copper and zinc on survival, whole body residues, and histopathology of mummichog, Fundulus heteroclitus (L.), were investigated in synthetic sea water at 20‰ salinity and 20°C. Mixtures of Cu²⁺ and Zn²⁺ as indicated by 96 h bioassay studies produced more deaths than expected on the basis of toxicities of individual components. Concentrations of Cd²⁺ not ordinarily lethal exerted a negative effect on survival of fish intoxicated by salts of copper, zinc, or both.
Atomic absorption determinations of Cd, Cu, and Zn residues in mummichog which survived 96 h exposures to each of these toxicants provided useful indices of total body burdens for these metals. Residues from survivors held in mixtures, especially Cd²⁺ and Zn²⁺ mixtures, did not conform to patterns observed for single elements. Whole body aggregates of Cd, Cu, and Zn from dead mummichogs were of limited worth owing to possible accumulation of these metals from the medium after death.
Renal and lateral line canal lesions were noted in all fish subjected to copper concentrations of 1 mg/1 and higher. Renal lesions observed in fish immersed in mixtures of Cu²⁺ and Cd²⁺ assumed a damage pattern characteristic of Cd²⁺; with mixtures of Cu²⁺ and Zn²⁺, lesion were typical of Cu²⁺-induced damage. Lesions induced in lateral line epithelium by Cu²⁺ were not affected by either Cd²⁺ or Zn²⁺. Epithelia lining the oral cavity were necrotized by the caustic action of high levels of Zn²⁺ (60 mg/1) and of Cu²⁺ (8 mg/1).     

The interactions of water hardness and pH with the acute toxicity of zinc to the brown trout, Salmo trutta L.

Exposure of brown trout, Salmo trutta , to zinc under continuous flow conditions over 96 h showed that both water hardness and pH exert major influences on the toxicity of the metal. 96-h LC50 values for total zinc ranged from <0.14mg 1⁻¹ in alkaline soft water (pH 8; lOmg 1⁻¹ as CaCO₃) to 3.20 mg 1⁻¹ in acidic hard water (pH 5; 204 mg 1⁻¹ as CaCO₃). A variable reduction in zinc toxicity in hard water compared with soft water over the pH range 4–9 was attributed to high external calcium. Zinc toxicity was positively correlated with decreasing acidity over the pH range 5–7, the metal being most toxic at pH 8–9 where metal complexes predominate. Below pH 5 metal toxicity also increased, irrespective of hardness. Water hardness and pH interacted with zinc toxicity in a complex manner, apparently dependent on physical and chemical transformations of the metal, and as changes in uptake. detoxification and excretion by the fish.     

Cadmium exposure of rainbow trout, Salmo gairdneri Richardson: effects on immune functions

Differential leucocyte counts, phagocytosis, humoral antibody response and the in vitro blasto-genetic response to mitogens (lipopolysaccharide and Concanavalin A) and to an antigen ( Vibrio anguillarum ) were studied in rainbow trout exposed to 0,0.7 or 3.6 μg Cd 1⁻¹ for 12 weeks.
Although the fish did not exhibit any clinical or histological changes, cadmium exposure was found to affect two of the immune parameters measured. The cellular response of fish immunized with V. anguillarum to the homologous antigen was significantly lower for splenocytes obtained from fish exposed to cadmium for 9 weeks (3.6 μg Cd 1⁻¹ group) than for splenocytes obtained from non-exposed fish. Conversely, the humoral antibody response to V. anguillarum O-antigen was higher in the 3.6 μg Cd 1⁻¹ group than in the non-exposed group. Protective immunity of fish vaccinated against V. anguillarum was equally as good in the cadmium-exposed group as in the non-exposed group. No cadmium-induced changes in differential leucocyte counts or in the proportions of phagocytic cells were observed.     

An in situ estimate of daily food consumption and alimentary canal evacuation rates of common carp, Cyprinus carpio L.

Daily food consumption and alimentary canal evacuation rates for common carp, Cyprinus carpio L., were estimated at 4-week intervals from April to October 1979. Both the rate of food evacuation and its relationship to temperature were described by an exponential function. Gut evacuation rates ranged from 6.0% of gut contents per hour at 9°C to 32.7% h⁻¹ at 26.5°C. Temperature accounted for 72–91% of the variation in gut evacuation rate among months. Daily food consumption varied from a low of 3.87 mg g⁻¹ of fish wet wt per day in April at 14°C (0.39% of wet body wt) to a high of 40.75 mg g⁻¹ (4.08%) in August at 26.5°C. Although a greater quantity of food appeared to be consumed during the morning in spring and early summer and a greater amount in the evening after July, there was not a statistically significant time of day for peak feeding activity. The field method described here seemed to provide reasonable estimates of food consumption compared with other field and laboratory studies.     

The effect of dissolved oxygen and salinity on the toxicity of ammonia to smolts of salmon, Salmo salar L.

The survival of Atlantic salmon smolts on exposure to constant concentrations of ammonia has been measured under laboratory conditions. At concentrations of dissolved oxygen close to the air-saturation value, the 24-h LC50 of un-ionised ammonia is 0.15 mg NH₃1⁻¹ in fresh water (hardness 264 mg 1⁻¹ as CaCO₃) and 0.3 mg NH₃1⁻¹ in 30% sea water; at concentrations of dissolved oxygen of 3.5 mg 1⁻¹ in fresh water and 3.1 mg 1⁻¹ in 30% sea water, the 24-h LC50 is 0.09 mg NH₃ 1⁻¹ and 0.12 mg NH₃ 1⁻¹ respectively; for fish acclimated for 1 day to a concentration of ammonia close to the 24-h median for un-acclimated fish, the median is increased between 38 and 79%, depending on test conditions.     

Effects of plant water deficit on the daily carbon balance of leaves of cacao seedlings

The daily carbon balance of individual source leaves of Theobroma cacao L. seedlings was measured at 2- to 3-day intervals during a 19-day period of increasing plant water deficit and during an 8-day period of recovery following rewatering. In each case, responses of stressed seedlings were compared to those of irrigated controls. Leaves of irrigated cacao seedlings assimilated approximately 41 mg carbohydrate dm^-2 during 12-h photoperiods, and exported an average of 34 mg carbohydrate dm^-2 during 24-h measurement cycles. The rate of carbon export from cacao leaves was sharply reduced as leaf water potential (ψ) declined between -0.8 and -2.0 MPa. Further, the rate of export was closely associated with the net assimilation rate (A), with export capacity being severely reduced as A fell to near zero. Net accumulation of dry matter occurred as long as A remained greater than approximately 20 mg carbohydrate dm^-2 over the 12-h photoperiod, but at lower assimilation rates, export exceeded concomitant assimilation. Carbon export continued at the expense of leaf carbon reserves as photoassimilation fell to near zero during periods of severe water stress (ψ < -2.0 MPa). Night respiration rate was independent of plant water status.     

Zinc binding to the gills of rainbow trout: the effect of long-term exposure to sublethal zinc

The effects of sublethal waterborne Zn (2·28 μmol l⁻¹) on Zn binding kinetics to the apical gill surface were studied in juvenile rainbow trout ( Oncorhynchus mykiss ). Two separate radiotracer techniques were employed to ascertain this information. First, in vitro binding kinetic experiments were performed at extremely elevated zinc concentrations (up to 20 mmol l⁻¹) to measure relatively low-affinity binding sites at the gill epithelium. There were no differences in Zn binding parameters ( K_m and B _max) for fish sublethally exposed to Zn for 21 days and their simultaneous controls. Nevertheless, Ca did have an increased inhibitory effect on Zn binding in Zn-exposed fish suggesting that the anionic groups on the gill epithelium of these fish had been altered in some manner. Additionally, in vivo Zn binding kinetics were investigated using environmentally relevant waterborne Zn concentrations (low μmol l⁻¹ range) to isolate high-affinity Zn binding sites (Ca transporters). No appreciable alterations in the Km and B _max values for Zn binding were seen between the Zn-exposed group and its simultaneous control following 15 days of exposure. Furthermore, no significant differences in CC morphometry were observed between treatments. Despite these lack of treatment effects, there were temporal alterations in K_m , B _max and CC fractional surface area in both groups. It is proposed that these fluctuations are controlled by hormonal factors (such as stanniocalcin), believed to play a role in Ca influx.     

Cardiorespiratory status of triploid brown trout during swimming at two acclimation temperatures

At 14° C, standard metabolic rate (75·1 mg O₂ h⁻¹ kg⁻¹), routine metabolic rate (108.8 mg O₂ h⁻¹ kg⁻¹), active metabolic rate ( c . 380 mg O₂ h⁻¹ kg⁻¹), critical swimming speed (U_crit 1·7 BL s⁻¹), heart rate 47 min⁻¹), dorsal aortic pressure (3·2 kPa) and ventilation frequency (63 min⁻¹) for triploid brown trout Salmo trutta were within the ranges reported for diploid brown trout and other salmonids at the same temperature. During prolonged swimming ( c . 80% U _crit), cardiac output increased by 2·3-fold due to increases in heart rate (1·8-fold) and stroke volume (1·2-fold). At 18° C, although standard and routine metabolic rates, as well as resting heart rate and ventilation frequency increased significantly, active metabolic rate and certain cardiorespiratory variables during exercise did not differ from those values for fish acclimated to 14° C. As a result, factorial metabolic scope was reduced (2·93-fold at 18° C v . 5·13-fold at 14° C). Therefore, it is concluded that cardiorespiratory performance in triploid brown trout was not unusual at 18° C, but that reduced factorial metabolic scope may be a contributing factor to the mortality observed in triploid brown trout at temperatures near 18° C.     

Cortisol stimulates hypo-osmoregulatory ability in Atlantic salmon, Salmo salar L.

Hypo-osmoregulatory ability in juvenile Atlantic salmon, Sulrno salur L., was improved by cortisol treatment. Implantation of a vegetable shortening pellet containing cortisol (50 mg kg⁻¹) resulted in elevated plasma cortisol titres. Maximum cortisol levels (160–170ngml⁻¹) were observed at days 6 and 12 after the implantation and dropped significantly by day 55. Cortisol-implanted fish in fresh water developed a twofold increase in gill Na⁺/K⁺-ATPase activity at days 6 and 12, and a threefold increase by day 55. Intestinal mucosa Na⁺/K⁺-ATPase activity was not affected by cortisol. Cortisol-implanted fish exposed to 28 ppt sea water for 48 h tended to show an improved ability to regulate their plasma osmolarity and reduce their ionic load. The osmo-regulatory ability attained at days 12 and 55 was further evaluated by exposing fish to 37 ppt sea water for 96 h. While all the control fish died relatively early in these tests, cortisol-implanted fish showed a clear reduction in their mortality rate. These results indicate that cortisol can induce biochemical and organismal changes during winter months that typify preadaptive events normally occurring in the spring.     

Mortality patterns in trout (Salmo trutta and S. gairdneri) exposed to an anionic detergent in relation to concentration and mechanisms of toxic action

SUMMARY. Histological and electron microscopical study of the gills of fish poisoned by the anionic detergent sodium lauryl sulphate indicates that the nature of the toxic action changes at a concentration of about 120 mg I⁻¹. Toxicity tests were conducted to determine whether this change could be detected in the test data, using brown trout ( Salmo trutta ) and rainbow trout ( S. gairdneri ).
Toxicity curves showed no indication of the change in toxic action, but at concentrations of 120 and 130 mg I⁻¹ of sodium lauryl sulphate 'split' probit lines occurred, and the lines for 150 mg I⁻¹ consistently differed in slope from other lines.     

Limnochemical and phytoplankton studies on Nyumba ya Mungu reservoir, Tanzania

This publication reports on the chemical and phytoplankton aspects of a three month biological survey of Nyumba ya Mungu reservoir, Tanzania.
The lake had a total salt concentration of about 8 meq dm^-3 and specific conductivity of 900 μS cm^-1. The main salt in solution was sodium bicarbonate, and when the lake level fell it sometimes formed a crust on the exposed shore. The concentrations of major nutrient ions were probably not limiting to algal growth and good nutrient replenishment was provided by the two inflow rivers.
Nyumba ya Mungu supports a rich phytoplanton dominated by Melosira and blue-green algae. Acetone extractions gave chlorophyll 'a' concentrations ranging from 20–40 mgm^-3 in the vertical profiles, and light and dark bottle experiments indicated a maximum gross photosynthesis of about 800 mg O₂ m^-3h^-1. There was evidence of chemical stratification in the open water and diurnal stratification in the sheltered bays, but discontinuities were short-lived and the Trade Winds ensured regular mixing of the water. The lake appeared to have a higher primary production than the larger Rift Valley lakes and it was considered that this level of production would continue.     

The effect of calcium concentration on the toxicity of copper, lead and zinc to yolk-sac fry of brown trout, Salmo trutta L., in soft, acid water

Yolk-sac fry of brown trout were exposed to three levels of single trace metals (Cu, 20,40,80 nmol 1^-1; Pb, 12·5,25,50 nmol 1^-1; Zn, 75,150,300 nmol 1^-1) typical of concentrations reported for acid soft waters, in flowing, artificial, soft water media maintained at pH 4·5 and [Ca] of 20 or 200 μmol 1^-1for 30 days.
Mortalities were high in fry subjected to all levels of the three trace metals at [Ca] 20 μmol 1^-1, with 80% of the total deaths occurring between days 11 and 15 of the experiment. 25% mortality occurred at low [Ca] and pH 4·5 in the absence of trace metals, with only one death recorded at [Ca] 200 μmol1^-1'(Cu, 80 nmol 1^-1). At high [Ca] all three levels of Cu and Pb impaired net Na and K uptake; Cu was the only metal to reduce the uptake of Ca. The Zn treatments had no significant effect on mineral uptake. Calcification of centra was reduced by all three Cu treatments at [Ca] 200 μmol 1^-1. The lowest Zn concentration (75 nmol 1^-1) was the only other treatment to impair skeletal development. In the absence of trace metals, low [Ca] significantly reduced Ca, Na and K uptake, skeletal calcification and dry mass at pH 4·5.
The deleterious effects of low Cu, Pb and Zn concentrations at low pH and low [Ca], and the ameliorative effect of higher ambient [Ca], are discussed in relation to fishery status in soft, acid waters.     

Some effects of low pH and calcium on the growth and tissue mineral content of yearling brown trout, Salmo trutta

Yearling brown trout, Salmo trutta , were exposed to low mineral content water (nominal concentrations of 20μmol 1⁻¹ magnesium, 7.7 μmol 1⁻¹ potassium, 44 μmol 1⁻¹ sodium) over a pH range of 4.0–5.2 with ambient calcium concentrations of 2.5–60 μmol 1⁻¹. All fish died at pH 4.0 and 4.2 irrespective of ambient calcium concentration and also at pH 4.4 with only 2–3 μmol 1 ⁻¹ calcium (that is calcium-free water except for that leached from the diet or excreted by the fish). Good growth rates were obtained over the remaining treatments which extended down to pH 4.4 with as little as 7 μmol 1⁻¹ calcium. When starved, weight loss was inversely correlated with pH. Effects on plasma chloride, percentage dry weight and calcium, potassium sodium, and phosphorus contents of skin, muscle and bone tissue were also investigated. These demonstrated pH effects on mineral metabolism in starved fish, but no effects were detected in fed fish.     

Effect of acute and chronic ammonia and nitrite exposure on oxygen consumption and growth of juvenile big bellied seahorse

Juvenile big bellied seahorse Hippocampus abdominalis were exposed acutely and chronically to elevated ammonia and nitrite {24 h exposure: 0·01, 5·0, 10·1, 14·8 and 19·9 mg l⁻¹ total ammonia-nitrogen [TA-N] and <0·001, 74·4, 99·2 and 123·6 mg l⁻¹ [NO₂-N] nitrite-nitrogen and 35 days exposure: 0·11, 0·55, 1·67 and 3·07 mg l⁻¹ TAN and <0·001, 0·92, 4·67 and 9·10 mg NO₂-N l⁻¹}. Significant ( P <0·001) increases in oxygen consumption rate and ventilation frequency occurred at 14·8, 19·9 mg l⁻¹ TA-N and 99·2, 123·6 mg l⁻¹ NO₂-N for acutely exposed fish. Oxygen consumption rate was significantly ( P <0·05) elevated at 1·67 and 3·07 mg l⁻¹ TA-N in chronically treated fish and ventilation frequency increased significantly ( P <0·05) at 0·55, 1·67, 3·07 mg l⁻¹ TA-N and 4·59, 9·10 mg l⁻¹ NO₂-N. There were no significant differences in growth between controls and ammonia exposed fish. Mortalities occurred at 14·8, 19·9 mg l⁻¹ TA-N.     

Episodic exposure to acid and aluminium in soft water: survival and recovery of brown trout,Salmo trutta L.

Yolk-sac fry, swim-up fry and 1–2 yr juveniles of brown trout, Sulmo trutta L., were exposed to episodes of aluminium and low pH, maximum aluminium concentration 12 μmol l⁻¹ (323 μg l⁻¹), minimum pH 4.5, total duration up to 54 h (yolk-sac fry) or up to 78 h (swim-up fry and juveniles), in an artificial soft water medium, [Ca] 20 μmol l⁻¹ (0–8 mg l⁻¹) (nominal baseline: pH 5.6, zero aluminium concentration). Yolk-sac fry mortality was nil or very low. A marked increase in susceptibility, with high mortalities, occurred when the yolk was fully absorbed. Mortality of juveniles exposed to two successive episodes was lower than would have been expected on the basis of comparisons with mortalities in single episodes, and mortality declined as the interval between the two episodes was increased. Disturbance of sodium, potassium or calcium balance or gill damage in surviving yolk-sac fry or juveniles was still evident 5 to 6 days after the end of a single episode.     

Evaluation and Potential Health Hazard of Selected Metals in Water,Sediments, and Fish from the Gomti River

The health hazard associated with the consumption of fish from the Gomti River in India, contaminated with the heavy metals Cr, Cu, Mn, Ni, Pb, and Zn was assessed in terms of target hazard quotients (THQs). The concentrations of metals (mg kg^?1, wet weight basis) in the muscle tissues of different fish species Mastacembelus puncalus, Clupisona garua, Cyrinous carpio, Botia lochachata, Channa punctatus, Heteropneustise fossilis, Puntius sofore, and Clarious batrachus ranged as follows: Cr (2.2–21.4), Cu (0.3–14.3), Mn (2.3–5.5), Ni (0.5–10.9), Pb (1.0–3.9), and Zn (12.3–46.9). The accumulation of metals in fish muscle tissue was in the order: Zn > Cr > Ni > Mn > Cu > Pb. THQs indicated a potential health hazard to children due to the consumption of fish contaminated with Ni and Pb; their THQs were greater than 1 for almost all fish species except for Ni in C. garua (THQ, 0.07) and C. carpio (THQ, 0.90). For adults, insignificant health hazard was associated with THQs less than 1 for all metals in the different fish species, but long-term exposure to these metals and subsequent bioaccumulation in the body may require additional investigation.