Marjan Diricx

Universiti Malaysia Terengganu, Tringano, Terengganu, Malaysia

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Publications (7)21.22 Total impact

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    ABSTRACT: Ammonia is an environmental pollutant that is toxic to all aquatic animals. The toxic effects of ammonia can be modulated by other physiological processes such as feeding and swimming. In this study, we wanted to examine these modulating effects in common carp (Cyprinus carpio). Fish were either fed (2% body weight) or starved (unfed for seven days prior to the sampling), and swimming at a sustainable, routine swimming speed or swum to exhaustion, while being exposed chronically (up to 28 days) to high environmental ammonia (HEA, 1 mg/L ~ 58.8 µmol/L as NH4Cl at pH 7.9). Swimming performance (critical swimming speed, Ucrit) and metabolic responses such as oxygen consumption rate (MO2), ammonia excretion rate (Jamm), ammonia quotient, liver and muscle energy budget (glycogen, lipid and protein), plasma ammonia and lactate, as well as plasma ion concentrations (Na+, Cl-, K+ and Ca2+) were investigated in order to understand metabolic and iono-regulatory consequences of the experimental conditions. Cortisol plays an important role in stress and in both the regulation of energy and the ion homeostasis, therefore plasma cortisol was measured. Results show that during HEA, Jamm was elevated to a larger extent in fed fish and they were able to excrete much more efficiently than the starved fish. Consequently, the build-up of ammonia in plasma of HEA exposed fed fish was much slower. MO2 increased considerably in fed fish after exposure to HEA and was further intensified during exercise. During exposure to HEA, the level of cortisol in plasma augmented in both the feeding regimes, but the effect of HEA was more pronounced in starved fish. Energy stores dropped for both fed and the starved fish with the progression of the exposure period and further declined when swimming to exhaustion. Overall, fed fish were less affected by HEA than starved fish, and although exercise exacerbated the toxic effect in both feeding treatments, this was more pronounced in starved fish. This suggests that fish become more vulnerable to external ammonia during exercise, and feeding protects the fish against the adverse effects of high ammonia and exercise.
    Aquatic Toxicology 08/2013; · 3.73 Impact Factor
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    ABSTRACT: Feeding and swimming can influence ion balance in fish. Therefore we investigated their impact on ionoregulation and its hormonal control in goldfish and common carp. As expected due to the osmorespiratory compromise, exhaustive swimming induced increases in gill Na(+)/K(+) ATPase (NKA) activity in both species, resulting in stable levels of plasma ions. In contrast to our expectations, this only occurred in fed fish and feeding itself increased NKA activity, especially in carp. Fasting fish were able to maintain ion balance without increasing NKA activity, we propose that the increase in NKA activity is related to ammonia excretion rather than ion uptake per se. In goldfish, this increase in NKA activity coincided with a cortisol elevation while no significant change was found in carp. In goldfish, high conversion of plasma T(4) to T(3) was found in both fed and fasted fish resulting in low T(4)/T(3) ratios, which increased slightly due to exhaustive swimming. In starved carp the conversion seemed much less efficient, and high T(4)/T(3) ratios were observed. We propose that thyroid hormone regulation in carp was more related to its role in energy metabolism rather than ionoregulation. The present research showed that both species, whether fed or fasted, are able to sufficiently adapt their osmorepiratory strategy to minimise ions losses while maintaining gas exchange under exhaustive swimming.
    Comparative biochemistry and physiology. Part A, Molecular & integrative physiology 02/2013; · 2.20 Impact Factor
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    ABSTRACT: Fish need to balance their energy use between digestion and other activities, and different metabolic compromises can be pursued. We examined the effects of fasting (7days) on metabolic strategies in goldfish and common carp at different swimming levels. Fasting had no significant effect on swimming performance (U(crit)) of either species. Feeding and swimming profoundly elevated total ammonia (T(amm)) excretion in both species. In fed goldfish, this resulted in increased ammonia quotients (AQ), and additionally plasma and tissue ammonia levels increased with swimming reflecting the importance of protein contribution for aerobic metabolism. In carp, AQ did not change since oxygen consumption (MO(2)) and T(amm) excretion followed the same trend. Plasma ammonia did not increase with swimming suggesting a balance between production and excretion rate except for fasted carp at U(crit). While both species relied on anaerobic metabolism during exhaustive swimming, carp also showed increased lactate levels during routine swimming. Fasting almost completely depleted glycogen stores in carp, but not in goldfish. Both species used liver protein for basal metabolism during fasting and muscle lipid during swimming. In goldfish, feeding metabolism was sacrificed to support swimming metabolism with similar MO(2) and U(crit) between fasted and fed fish, whereas in common carp feeding increased MO(2) at U(crit) to sustain feeding and swimming independently.
    Comparative biochemistry and physiology. Part A, Molecular & integrative physiology 08/2012; 163(3-4):327-35. · 2.20 Impact Factor
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    ABSTRACT: Waterborne ammonia has become a persistent pollutant of aquatic habitats. During certain periods (e.g. winter), food deprivation may occur simultaneously in natural water. Additionally, under such stressful circumstances, fish may be enforced to swim at a high speed in order to catch prey, avoid predators and so on. Consequently, fish need to cope with all these stressors by altering physiological processes which in turn are controlled by their genes. In this present study, toxicogenomic analyses using real time PCR was used to characterize expression patterns of potential biomarker genes controlling growth, ion regulation and stress responses in common carp subjected to elevated ammonia (1 mg/L; Flemish water quality guideline for surface water) following periods of feeding (2% body weight) and fasting (unfed for 7 days prior to sampling). Both feeding groups of fish were exposed to high environment ammonia (HEA) for 0 h (control), 3h, 12h, 1 day, 4 days, 10 days, 21 days and 28 days, and were sampled after performing swimming at different speeds (routine versus exhaustive). Results show that the activity and expression of Na(+)/K(+)-ATPase, an important branchial ion regulatory enzyme, was increased after 4-10 days of exposure. Effect of HEA was also evident on expression patterns of other ion-regulatory hormone and receptor genes; prolactin and cortisol receptor mRNA level(s) were down-regulated and up-regulated respectively after 4, 10 and 21 days. Starvation and exhaustive swimming, the additional challenges in present study significantly further enhanced the HEA effect on the expression of these two genes. mRNA transcript of growth regulating hormone and receptor genes such as Insulin-like growth factor I, growth hormone receptor, and the thyroid hormone receptor were reduced in response to HEA and the effect of ammonia was exacerbated in starved fish, with levels that were remarkably reduced compared to fed exposed fish. However, the expression of the growth hormone gene itself was up-regulated under the same conditions. Expression of somatolactin remained unaltered. Stress representative genes, cytochrome oxidase subunit 1 showed an up-regulation in response to HEA and starvation while the mRNA level of heat shock protein 70 was increased in response to all the three stressors. The expression kinetics of the studied genes could permit to develop a "molecular biomarker system" to identify the underlying physiological processes and impact of these stressors before effects at population level occur.
    Aquatic toxicology (Amsterdam, Netherlands) 06/2012; 122-123:93-105. · 3.12 Impact Factor
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    ABSTRACT: Due to eutrophication, high environmental ammonia (HEA) has become a frequent problem in aquatic environments, especially in agricultural or densely populated areas. During certain periods, e.g. winter, feed deprivation may occur simultaneously in natural waters. Additionally, under such stressful circumstances, fish may be enforced to swim at a high speed in order to catch prey, avoid predators and so on. Consequently, fish need to cope with all these stressors by altering physiological processes which in turn are controlled by genes expression. Therefore, in the present study, ammonia toxicity was tested in function of nutrient status (fed versus starved) and swimming performance activity (routine versus exhaustive). Goldfish, a relatively tolerant cyprinid, were exposed to HEA (1 mg/L; Flemish water quality guideline for surface water) for a period of 3 h, 12 h, 1 day, 4 days, 10 days, 21 days and 28 days and were either fed (2% body weight) or starved (kept unfed for 7 days prior to sampling). Results showed that the activity of Na⁺/K⁺-ATPase in the gills was stimulated by HEA and disturbance in ion balance was obvious with increases in plasma [Na⁺], [Cl⁻] and [Ca²⁺] after prolonged exposure. Additionally, osmoregulation and metabolism controlling hormones like cortisol and thyroid hormones (T3 and T4) were investigated to understand adaptive responses. The expression kinetics of growth, stress and osmo-regulatory representative genes such as Insulin-like growth factor 1 (IGF-I), growth hormone receptor (GHR), thyroid hormone receptor β (THRβ), prolactin receptor (PRLR), cortisol receptor (CR) and Na⁺/K⁺-ATPase α(3) were examined. Overall effect of HEA was evident since Na⁺/K⁺-ATPase activity, plasma cortisol, Na⁺ and Ca²⁺ concentration, expression level of CR and Na⁺/K⁺-ATPase α₃ mRNA in fed and starved fish were increased. On the contrary, transcript level of PRLR was reduced after 4 days of HEA; additionally T3 level and expression of GHR, IGF-I and THRβ genes were decreased following 10-21 days of HEA. Starvation, the additional challenge in the present study, significantly increased plasma cortisol level and CR transcript level under HEA compared to the fed exposed and control fish. Furthermore, a remarkable reduction in T3 and mRNA levels of THRβ, IGF-I and GHR genes was observed under starvation. The toxic effects in both feeding treatments were exacerbated when imposed to exhaustive swimming with more pronounced effects in starved fish. This confirms that starvation makes fish more vulnerable to external ammonia, especially during exercise.
    Aquatic toxicology (Amsterdam, Netherlands) 03/2012; 114-115:153-64. · 3.12 Impact Factor
  • Aquatic Toxicology 01/2012; · 3.73 Impact Factor
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    ABSTRACT: This study aimed to elucidate the physiological effects of high environmental ammonia (HEA) following periods of feeding (2% body weight) and starvation (unfed for 7 days prior to sampling) in gold fish (Carassius auratus). Both groups of fish were exposed to HEA (1 mg/L; Flemish water quality guideline) for 0 h (control), 3 h, 12 h, 1 day, 4 days, 10 days, 21 days and 28 days. Measurements of weight gain (%), oxygen consumption (MO2), ammonia excretion rate, ammonia quotient (AQ), critical swimming speeds (Ucrit), plasma and muscle ammonia accumulation, plasma lactate, liver and muscle glycogen, lipid and protein content were done at various time intervals during the experimental periods. Overall, ammonia excretion rates, plasma ammonia accumulation and AQ were significantly affected by food regime in ammonia free water. HEA, the additional challenge in the present study, significantly altered all the studied parameters among fed and starved groups in days-dependent manner. Results show that weight gain (%), MO2, Ucrit, protein content in liver and muscle, and glycogen content in muscle among starved fish under HEA were considerably reduced compared to control and fed fish. Additionally a remarkable increase in plasma ammonia level, muscle ammonia, lactate accumulation and AQ was seen. However in fed fish, MO2, ammonia excretion rate, AQ and lactate level augmented after exposure to HEA. These results indicate that starved fish appeared more sensitive to HEA than fed fish. Furthermore, as expected, the toxic effect of ammonia exposure in both feeding treatments was exacerbated when imposed to exhaustive swimming (swum at 3/4th Ucrit). Such effects were more pronounced in starved fish. This suggests that starvation can instigate fish more vulnerable to external ammonia during exercise. Therefore, it was evident from our study that feeding ameliorates ammonia handling and reduces its toxicity during both routine and exhaustive swimming. Moreover, recovery was observed for some physiological parameters (e.g. MO2, ammonia excretion, Ucrit, plasma ammonia) during the last exposure periods (21-28 days) while for others (e.g. growth, tissue glycogen and protein content, muscle ammonia) effects only became apparent at this time. In the future, these results need to be considered in ecological context as fish in ammonia polluted may experience different phenomenon (starvation and exercise) simultaneously.
    Aquatic toxicology (Amsterdam, Netherlands) 11/2011; 109:33-46. · 3.12 Impact Factor