Differential effects of exposure to parasites and bacteria on stress response in turbot Scophthalmus maximus simultaneously stressed by low water depth

The stress response of turbot Scophthalmus maximus was evaluated in fish maintained 8 days under different water depths, normal (NWD, 30 cm depth, total water volume 40 l) or low (LWD, 5 cm depth, total water volume 10 l), in the additional presence of infection–infestation of two pathogens of this species. This was caused by intraperitoneal injection of sublethal doses of the bacterium Aeromonas salmonicida subsp. salmonicida or the parasite Philasterides dicentrarchi (Ciliophora:Scuticociliatida). The LWD conditions were stressful for fish, causing increased levels of cortisol in plasma, decreased levels of glycogen in liver and nicotinamide adenine dinucleotide phosphate (NADP) and increased activities of G6Pase and GSase. The presence of bacteria or parasites in fish under NWD resulted in increased cortisol levels in plasma whereas in liver, changes were of minor importance including decreased levels of lactate and GSase activity. The simultaneous presence of bacteria and parasites in fish under NWD resulted a sharp increase in the levels of cortisol in plasma and decreased levels of glucose. Decreased levels of glycogen and lactate and activities of GSase and glutathione reductase (GR), as well as increased activities of glucose‐6‐phosphate dehydrogenase (G6PDH), 6‐phosphogluconate dehydrogenase (6PGDH) and levels of nicotinamide adenine dinucleotide phosphate (NADPH) occurred in the same fish in liver. Finally, the presence of pathogens in S. maximus under stressful conditions elicited by LWD resulted in synergistic actions of both type of stressors in cortisol levels. In liver, the presence of bacteria or parasites induced a synergistic action on several variables such as decreased activities of G6Pase and GSase as well as increased levels of NADP and NADPH and increased activities of GPase, G6PDH and 6PGDH.     

Interactive effect of ammonia and crowding stress on ion-regulation and expression of immune-related genes in juvenile turbot (Scophthalmus maximus)

The present study was to investigate the interactive effect of ammonia and crowding stress on ion-regulation and expression of immune-related genes in juvenile turbot (Scophthalmus maximus). The fish were exposed to four total ammonia nitrogen (0, 5, 20, 40 mg/L TAN) and two stocking density. After 96 h of exposure, blood, gill, and liver samples were collected to measure biochemical parameters and mRNA levels of immune-related genes. The results showed that co-exposure to high TAN (20 and 40 mg/L) and high density significantly increased plasma sodium (Na⁺), gill Na⁺/K⁺-ATPase activity and mRNA levels. Following individual and combined exposure to high TAN and high density, the heat shock protein 70 (HSP 70), HSP 90, tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) genes expression were obviously higher than their control. Conversely, the lysozyme (LZM) and hepcidin mRNA levels were down-regulated in liver of fish exposed to high TAN alone and combination of high TAN-density. Moreover, glutathione S-transferase (GST) mRNA levels significantly increased in treatments with individual high TAN and high density, but decreased in high TAN-density co-exposed fish. Overall, ion homeostasis and immune status were adversely influenced in high exposed turbot under high density.     

The ageing phenomenon of turbot spermatozoa: effects on morphology, motility and concentration, intracellular ATP content, fertilization, and storage capacities

The deleterious effect of the ageing phenomenon of turbot spermatozoa was investigated in relation to the sampling date. Spermatozoa with a low or highly condensed chromatin and a middle piece containing numerous or a few vesicles were observed simultaneously 80 and 47 days before the beginning of the spawning period of the females. The middle piece of spermatozoa contained few vesicles, 39 days after the end of the reproductive period. At the same date, some spermatozoa appeared in which the plasma membrane was broken. Sperm motility, assessed just after collection in terms of arbitrary motility scores from 0 to 5, was significantly increased both at 10 and 60 s post-activation, for samples collected 18 days after, 25 days before and 9 days after the beginning of the spawning period of the females, respectively compared to samples collected 6 days before, 55 days after and 88 days after the end of this period. A lower short-term storage capacity was recorded at 10 and 60 s post-activation for sperm samples collected 6 days before and 88 days after the end of the reproductive period, respectively compared to 18 days and 9 days after the beginning of the spawning period. At 60 s post-activation, a higher motility of thawed spermatozoa was observed for samples collected 5 days before the beginning of the spawning period (motility recovery index: 86.4 ± 19.4%) compared to 71 days after the end of this period (55.0 ± 12.0%). The fertilizing capacity of sperm samples collected 61 days after the end of the spawning season (66.1 ± 14.6%) was significantly lower than that recorded for samples collected 34 days after the beginning of the spawning period (75.2±9.6%). On the contrary, there was no significant decrease in endogenous ATP content (31 days after the beginning of the spawning period, 14.53 ± 0.84; 48 days after the end of this period, 10.75 ± 5.26 nmol 10^{? 8} spermatozoa). Furthermore, sperm concentration significantly increased between the same dates (respectively 3.3 ± 0.8–9.4±4.8×10⁹ spermatozoa ml^?1).     

The interaction of temperature and fish size on growth of juvenile turbot

Growth rate of tagged juvenile turbot was significantly influenced by the interaction of temperature and fish size. The results suggest the optimum temperature for growth of juvenile turbot in the size range 25–75 g is between 16 and 19°C. Optimal temperature for growth decreased rapidly with increasing size, and is between 13 and 16°C for 100 g turbot. Although individual growth rates varied highly at all times within the temperature treatments, significant size rank correlations were maintained during the experimental period. The study confirms that turbot exhibit ontogenetic variation in temperature optimum, which might partly explain different spatial distribution of juvenile and adult turbot in ocean waters.     

The combined effects of chlorine and ammonia on litter breakdown in outdoor experimental streams

The response of Potamogeton crispus L. breakdown to controlled doses of different levels of chlorine and chlorine + ammonia was investigated over two years in outdoor experimental streams. In 1985, downstream riffles of 2 streams were dosed (observed in-stream concentrations) at ca. 10 μg/L Total Residual Chlorine (TRC), one stream at 64 μg/L TRC and one stream at 230 μg/L TRC. Two control streams were not dosed and the upstream riffles of each stream served as within stream controls. In 1986, the downstream riffle of one stream was dosed at 70 μg/L TRC and a second stream was dosed at 200 μg/L TRC. Four streams were also dosed with 2.5 mg/L NH₃-N: one stream with no chlorine, one stream with ca. 10 μg/L TRC, one with 56 μg/L TRC, and one with 150 μg/L TRC. A seventh stream was dosed for 2 h at 2000 μg/L TRC and 2.5 mg/L ammonia and then allowed to recover (recovery stream). Each year, litter decomposition (degree day k values) was measured during two 35 day trials (Jun–Jul and Aug–Sep). In 1985, when streams were dosed with chlorine alone, decomposition was significantly reduced with the high (230 μg/L TRC) chlorine dose. Downstream decomposition was 27% (Jun–Jul) and 59% (Aug–Sep) of the upstream (control) rate. No other chlorine effects were found during this period. In Jun–Jul 1986, there was significantly lower decomposition in the downstream dosed sites of the 200 μg/L TRC alone stream, the 146 μg/L TRC + ammonia stream and the recovery stream; downstream decay rates were (respectively) 56%, 42% and 64% of the upstream control sites. No other up-down pairs were different in July 1986. In Aug–Sep, all three streams with chlorine + ammonia (6, 56 and 146 μg/L TRC + 2,5 mg/L ammonia) and the 70 μg/L TRC alone stream had significantly lower decomposition rates in the downstream dosed sites. For these streams, downstream decay rates ranged from 46% (high chlorine + ammonia) to 73% (low chlorine + ammonia) of the upstream control rates. No other up-down pairs were different during this trial. Up and downstream sites of the stream dosed with 2.5 mg/L ammonia alone were nearly identical for both trials (< 3% difference). These results indicate that TRC at less than 250 μg/L can significantly reduce litter decomposition and strongly suggest that addition of ammonia to chlorinated water can increase the toxic effect of chlorine. currently at the Department of Fisheries and Wildlife currently at the Department of Fisheries and Wildlife     

Stochastic simulation of size variation in turbot: possible causes analysed with an individual-based model

Causes of size variation in a population of juvenile turbot were studied using an individual based model (IBM). Each simulation started with 800 (divided into eight groups of 100 each) 120-day-old (posthatch) juveniles and was run for 140 days, and the data gained from model simulations compared directly with the result of a laboratory study with size-graded turbot. Stochastic growth with memory, which was included in the models as an individual genetical growth rate variation, is important in explaining size variation, and the combination between individual genetic growth rate and social interactions related to size-dependent hierarchies also contributes to size variation. The use of size-dependent growth rate alone fails to explain size variation, and is of little value in predicting size variation in turbot culture. Further, the results indicate formation of different types of size hierarchies for different sizes of juvenile turbot.     

摄食水平和投喂频率对大菱鲆幼鱼生长及生化成分的影响

以大菱鲆幼鱼(7.56±0.03 g)为试验对象,在水温17～19℃下,设计了4个摄食水平梯度(1.0％、1.5％、2.0％和饱食)和3个投喂频率梯度(1、2和3次·d-1),探讨了不同摄食水平和投喂频率对其生长及生化成分的影响.结果表明:随着摄食水平的升高,大菱鲆幼鱼的终末体重、相对增重率和摄食率显著升高(P＜0.05);特定生长率随摄食水平的增加呈线性升高;湿重、干重、蛋白质和能量转化效率出现先升高后降低的趋势;摄食水平对鱼体的体成分和组间表观消化率无显著影响(P＞0.05);不同投喂频率下,终末体重、相对增重率、特定生长率及转化效率呈先升高再降低的趋势,在2次·d-1时取得最大值;投喂频率对鱼体的体成分和组间表观消化率无显著影响(P＞O.05).     

Effects of salinity on the ionic balance and growth of juvenile turbot

The effects of salinity changes (27, 19 and 10‰) on seawater-adapted juvenile turbot were studied on their plasma osmolarity and ion concentrations, on oxygen consumption, on gill Na⁺,K⁺-ATPase activity after 3 months and on growth parameters. All plasma concentrations (except chloride) were unchanged, suggesting that fish were well adapted to their environment. Oxygen consumption was significantly decreased in the 19 and 10‰ groups, where fish weighed significantly more 105 days after transfer than fish maintained in sea water. These results, and the fact that apparent food conversion rates were lower in a diluted environment, suggest that on a long term schedule growth conditions could be improved by adaptation to brackish waters (salinities between 10 and 19‰). The effects of transfer from sea water to 27, 19, 10 and 5‰ were also followed during the first 3 weeks. With salinity 10‰ a steady state was reached on day 21 with all plasma values within the same range. The significant differences observed in osmolarity, plasma ion concentrations and Na⁺,K⁺-ATPase activity 3 weeks after transfer of juveniles to 5‰ salinity, compared with transfers in higher salinities, suggest that there is a threshold of acclimation of turbot to a hypotonic environment.     

Effect of temperature and dissolved oxygen concentration on the metabolic rate of the turbot and the relationship between metabolic scope and feeding demand

Turbot Scophthalmus maximus maximum oxygen uptake following feeding and exhaustive exercise increased from 107 mg O2 kg⁻¹ h⁻¹ at 6° C to c . 218 mg O2 kg⁻¹ h⁻¹ at 18° C, then increased slightly from 18 to 22° C to 224 mg O2 kg⁻¹ h⁻¹. Standard oxygen uptake increased exponentially as a function of temperature from 11 mg O2 kg ⁻¹ h⁻¹ at 6° C to 66 mg O2 kg⁻¹ h⁻¹ at 22° C. Gradual reduction in oxygen concentration to 87–90% air saturation at 6, 10. 18° C and <80% at 14 and 22° C limited the maximum metabolic rate but, supersaturation (>100% saturation) had little effect. Metabolic scope attained a maximum of 176 mg O₂ kg⁻¹ h⁻¹ at 18° C. Interpolation of the results showed that this value changed little between 16 and 20° C. It is suggested that this temperature range is optimal for turbot of c . 500 g. A comparison with a previous study on feeding demand in intensive farming conditions showed a linear relationship between appetite and metabolic scope. It is concluded that the ability of a fish to supply energy (including the energy requirement of digestive metabolism) above a standard level is a limiting factor in the manifestation of its feeding demand.     

Chronic toxicity of ammonia to juvenile gilthead seabream Sparus aurata and related histopathological effects

Effect of 20 days exposure of juvenile gilthead seabream ( Sparus aurata ) to elevated levels of ammonia on growth and survival was examined in a continuous flow system. Suppressed growth and reduced survival were observed at concentrations of 8.2 and 13 mg l⁻¹ total ammonia-N (0.5 and 0.7 mg l⁻¹ un-ionized ammonia-N, respectively) and higher. The maximum acceptable toxic concentration (MATC) for growth was between 4.8 to 8.2 mg l⁻¹ total ammonia-N (0.3 and 0.5 mg l⁻¹ un-ionized ammonia-N, respectively). Fish exposed to high ammonia levels (13 mg l⁻¹ total ammonia-N, 0.7 mg l⁻¹ un-ionized ammonia-N) displayed clear signs of liver pathology. Existing evidence suggests that S. aurata is less sensitive to ammonia than other reported marine and freshwater fish.
Under certain conditions ammonia concentration in the intensive fish ponds in Eilat may exceed the no observed effect concentration for S. aurata .     

Microsatellite DNA variation in wild populations and farmed strains of turbot from Ireland and Norway: a preliminary study

Turbot Scophthalmus maximus is the focus of a rapidly expanding aquaculture industry, while at the same time wild catches appear to be in decline. As a preliminary investigation into the effects of hatchery rearing, genetic variation at three polymorphic microsatellite loci was assessed in two wild populations and two farmed strains of turbot, from Ireland and Norway. Although a considerable loss of rare alleles was observed in the Irish farmed strain, no statistically significant reductions were found in mean heterozygosity or allelic diversity in farmed strains compared to wild populations. Significant genetic heterogeneity was found between wild and farmed samples from each country but not between the two wild populations. Genetic differentiation between the farmed strains was presumed to be caused by drift in the hatcheries. The utility of these particular microsatellite loci in comparing these samples and the importance of molecular genetic testing of farmed strains is stressed.     

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.     

三种不同小球藻去除亚硝态氮和氨氮能力的研究

为了研究小球藻在不同质量浓度的氨氮和亚硝态氮环境下的去除能力与生长效果,从实验室挑选3株不同的小球藻CV315-1(Chlorella sp.)、CV315-2(Chlorella sorokiniana)和CV315-3(Chlorella pyrenoidosa)分别置于不同质量浓度的氨氮和亚硝态氮模拟污水中,在温...     

不同包膜控释尿素对农田土壤氨挥发的影响

为了探索包膜控释尿素土壤氨挥发损失规律特征和提高肥料氮素利用率,采用小麦玉米轮作田间试验,通过与普通尿素进行对比,运用土壤氨挥发原位测定方法——通气法系统研究了硫包膜和树脂包膜控释尿素的施用对小麦玉米轮作农田土壤氨挥发的影响.研究结果表明:在两种施氮量水平下(210 kg/hm2和300 kg/hm2),与普通尿素相比,硫包膜和树脂包膜控释尿素在小麦基肥期、小麦追肥期和玉米施肥期的施用均减少了土壤氨挥发的累积损失量,分别达35.1％-54.3％、59.6％-75.2％、65.6％-98.1％;有效降低了土壤氨挥发通量峰值且延迟其出现时间3-8 d,并能延缓土壤氨挥发主要阶段的时间分别为4-12 d、5-12 d.在小麦玉米轮作周年中,控释尿素土壤氨挥发累积损失量为28.39-43.35 kg/hm2,土壤氨挥发损失率为4.48％-5.63％,控释尿素时段土壤氨挥发通量比普通尿素降低了51.0％-70.8％;且树脂包膜控释尿素的施用降低小麦玉米轮作农田土壤氨挥发的效果优于硫包膜控释尿素.     

The influence of feeding regime on sexual maturation, fecundity and atresia in first-time spawning turbot

Exposure of adolescent turbot Scophthalmus maximus to low rations during vitellogenesis, covering the 4 months immediately prior to spawning, led to a drop of 70% in mean ovary weight, and was associated with poor growth of the vitellogenic oocytes or, in a third of cases, the absence of vitellogenic oocytes. Exposure to low rations during the recruitment of vitellogenic oocytes, 4–8 months prior to spawning, produced a more variable response. A few of these females produced well-developed vitellogenic oocytes, but in most, vitellogenic oocytes were absent or only poorly developed and in some cases there was a high incidence of atresia. Exposure to intermediate rations throughout oocyte recritment and vitellogenesis also depressed oocyte development. In maturing females the number of non-atretic vitellogenic oocytes (relative potential fecundity) averaged 998 per g of somatic body weight of fish at the start of the spawning season in mid-June. Maturation of the males was not significantly influenced by the dietary regime. The small size of the testes and the low levels of milt production, even in fish fed high rations immediately prior to spawning, showed that reproductive investment in males was much lower than in females.     

不同缓控释肥对鲜食玉米产量、品质及氨挥发的影响

采用田间试验,设置不施氮对照(CK)、常规施肥(U)、增效尿素(DU)、包膜尿素(CU)、缓控释掺混尿素(CDU)共5个处理,研究了常规施肥(240 kg N·hm^-2)和不同缓控释肥料一次性减量施用(180 kg N·hm^-2)对鲜食玉米产量、品质与土壤无机氮变化和氨挥发的影响。结果表明: U处理氨挥发总量最高,追肥是产生氨挥发损失的重要因素;与U处理相比,DU、CU、CDU处理氨挥发减排78%～81%。收获后U处理80～100 cm土层硝态氮浓度最高,为51.6 mg·kg^-1,氮淋溶风险较高,而DU、CU、CDU处理同土层硝态氮含量均较低,降低了淋溶风险。与U处理相比,减氮25%的3个缓控释肥处理没有减产,并增加了籽粒维生素C、可溶性糖和蛋白质含量;缓控释肥处理之间,DU处理的氮肥农学效率和经济效益最高。综上,减量施用新型缓控释肥可以实现鲜食玉米稳产提质,显著降低氨挥发损失和硝态氮淋失风险。与成本较高的树脂包膜控释肥相比,双效抑制剂增效肥(DU)成本低、制作便捷,是鲜食玉米专用肥的较好选择。     

Defences against ammonia toxicity in tropical air-breathing fishes exposed to high concentrations of environmental ammonia: a review

In the tropics, air-breathing fishes can be exposed to environmental ammonia when stranded in puddles of water during the dry season, during a stay inside a burrow, or after agricultural fertilization. At low concentrations of environmental ammonia, NH₃ excretion is impeded, as in aerial exposure, leading to the accumulation of endogenous ammonia. At high concentrations of environmental ammonia, which results in a reversed NH₃ partial pressure gradient (P_NH3), there is retention of endogenous ammonia and uptake of exogenous ammonia. In this review, several tropical air-breathing fishes (giant mudskipper, African catfish, oriental weatherloach, swamp eel, four-eyed sleeper, abehaze and slender African lungfish), which can tolerate high environmental ammonia exposure, are used as examples to demonstrate how eight different adaptations can be involved in defence against ammonia toxicity. Four of these adaptations deal with ammonia toxicity at branchial and/or epithelial surfaces: (1) active excretion of NH₄⁺; (2) lowering of environmental pH; (3) low NH₃ permeability of epithelial surfaces; and (4) volatilization of NH₃, while another four adaptations ameliorate ammonia toxicity at the cellular and subcellular levels: (5) high tolerance of ammonia at the cellular and subcellular levels; (6) reduction in ammonia production; (7) glutamine synthesis; and (8) urea synthesis. The responses of tropical air-breathing fishes to high environmental ammonia are determined apparently by behavioural adaptations and the nature of their natural environments.Communicated by I.D. Hume     

生物炭对农田土壤氨挥发的影响机制研究进展

降低土壤氨挥发量是农田生态系统中减少土壤氮素损失、提高氮肥利用率的关键途径之一。生物炭具有独特的理化性质,施入土壤后可改变土壤理化性状,影响土壤氮素循环,并对农田土壤中氨挥发产生重要的影响。本文首先介绍了稻田和旱田两种土地利用方式下农田氨挥发过程及其影响因素(气候条件、土壤环境、施肥管理等);其次,重点综述了生物炭对农田生态系统氨挥发影响的研究进展,并从物理吸附机制、气液平衡机制、生物化学过程调节机制等方面探讨了生物炭介入下农田土壤氨挥发的响应机制,认为土壤氨挥发减排的响应主要是基于生物炭表面含氧官能团对土壤NH₄⁺和NH₃的吸附作用及促进土壤硝化作用;而生物炭增加土壤氨挥发排放主要与生物炭提高土壤pH值和透气性、增强土壤有机氮矿化微生物活性有关。最后,对生物炭减少土壤氨挥发、提高氮肥利用率的研究方向进行了展望。     

The volatilization of ammonia from cattle urine applied to soils as influenced by soil properties

The amounts of ammonia volatilized, following the application of cattle urine to 22 soils, were measured in the laboratory during an incubation period of 10 days. The urine contained 12.0 g N dm^-3 and was applied to small columns of soil at a rate equivalent to 26.5 g N m^-2. The soils were from fields of both grassland and arable cultivation and varied widely in properties. Ammonia volatilization ranged from 6.8 to 41.3% of the total urinary N, with a mean value of 26.4%. The soil property most closely related to the extent of volatilization was cation exchange capacity (CEC), and this was so whether all 22 soils were considered together or whether the 14 grassland and 8 arable soils were considered separately. In general, the higher the CEC the less the amount of ammonia volatilized. However, for a given value of CEC, volatilization tended to be greater from a grassland than from an arable soil. The pH of a soil/urine mixture measured after 24 hours was also quite closely correlated with the amount of ammonia volatilized, but the initial pH and titratable acidity of the soil were poorly correlated with ammonia volatilization. ei]H Marschner ei]H Lambers