Protective effect of dietary curcumin in Anabas testudineus (Bloch) with a special note on DNA fragmentation assay on hepatocytes and micronucleus assay on erythrocytes in vivo

The present study was conducted to evaluate the safety of long-term dietary curcumin at doses 0.5 and 1 % in Anabas testudineus employing hematological and cytological techniques. The fish were fed with curcumin-supplemented feed for 6 months. Fine blood smears were prepared and subjected to three different staining techniques. The erythrocyte micronucleus frequency (MN) and the cytometric measurements of erythrocytes were determined. Blood from the control and treated fish was subjected to the assessment of several hematological parameters. Also, DNA fragmentation assay on hepatocytes was conducted. The results showed that hemoglobin content, RBC count and hematocrit increased in the curcumin-fed fish compared to control, whereas WBC count, platelet count, mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration were unaffected. WBC/RBC ratio was lower in the case of curcumin-treated fish. The cytometric measurements revealed no change in the erythrocytes and their nuclei after curcumin treatment. DNA fragmentation assay revealed intact DNA in curcumin-fed group, ruling out the possibility of curcumin-induced DNA damage. The positive control group showed a significant increase in MN frequency compared to negative control and curcumin-fed groups. In fact, the MN frequency decreased in 1 % curcumin-fed group compared to the negative control and 0.5 % curcumin groups. All these indicated a state of well-being of the curcumin-treated fish. Therefore, it is concluded that curcumin could be used as a safe feed ingredient to improve the growth of finfish in aquaculture.     

Oxidative metabolism in a teleost,Anabas testudineus Bloch: effect of testosterone and estradiol-17β on hepatic enzyme activities

Testosterone (T) administration to maleAnabas testudineus significantly stimulated the activities of cytochrome oxidase, -glycerophosphate dehydrogenase (-GPDH) and Mg²⁺ adenosine triphosphatase (Mg²⁺ ATPase) and inhibited lactate dehydrogenase (LDH), cytosolic and mitochondrial malate dehydrogenases (MDH). The activities of succinate dehydrogenase (SDH), glucose-6-phosphate dehydrogenase (G-6-PDH) and catalase were unaffected by testosterone treatment. Administration of estradiol-17 (E2) in female fish, significantly stimulated cytochrome oxidase activity, inhibited Mg²⁺ ATPase, SDH, catalase and cytosolic and mitochondrial MDH activity, and was without effect on other enzymes studied.The simultaneous injections of actinomycin D or chloramphenicol and T or E2 prevented the hormonal influence on hepatic enzyme activities. The present study demonstrates that inA. testudineus sex steroids influence hepatic oxidative metabolism by a mechanism sensitive to the action of inhibitors of protein synthesis.     

Modulation of digestive enzymes,GH, IGF-1 and IGF-2 genes in the teleost,Tilapia (<Emphasis Type="Italic">Oreochromis mossambicus</Emphasis>) by dietary curcumin

Aquaculture is faced with the challenges of the use of synthetic compounds as growth enhancers and the presence of several contaminants in water. These factors severely deteriorate the quality and quantity of aquaculture products. Phytochemicals play a major role by working as antioxidant agents of which curcumin has become the gold standard. Curcumin, from Curcuma longa shows a wide spectrum of biological activities which include anticancerous, antioxidant, anti-inflammatory, antibacterial, antiviral, antifungal, antidiabetic, antistress, hepatoprotective and gastroprotective effects. Curcumin in 0.5 and 1 % doses were given as feed additive to Oreochromis mossambicus for 35 days. After feeding trial, activities of digestive enzymes such as α-amylase, protease and lipase were analysed. There was a significant increase in the activities of α-amylase, protease and lipase with 0.5 and 1 % curcumin supplementation in feed. Real-time quantification of GH in brain, and IGF-1 and IGF-2 genes in muscle revealed that curcumin significantly increased the expression of these genes. This is the first study to report that curcumin supplementation at concentrations of 0.5 and 1 % in the feed improved the activities of digestive enzymes and also modulates the expression of GH in brain and growth factors such as IGF-1 and IGF-2 in muscle of O. mossambicus.     

Curcumin analogue inhibits lipid peroxidation in a freshwater teleost, <Emphasis Type="Italic">Anabas testudineus</Emphasis> (Bloch)—an in vitro and in vivo study

The effect of a synthetic curcumin analogue (salicylcurcumin) on fish lipid peroxidation was investigated in both in vitro and in vivo conditions using a teleost model Anabas testudineus (Bloch). Curcumin analogue inhibited the formation of lipid peroxidation products and thiobarbituric acid reactive substances (TBARS) content at the three concentrations (10⁻² M, 10⁻³ M and 10⁻⁴ M) in vitro. TBARS content was reduced by 80% in the liver and 68% in brain by the higher concentration of salicylcurcumin. For in vivo study, salicylcurcumin (0.5%) was supplemented along with the basal feed for a period of 60 days. It produced a 60% reduction in liver TBARS content. The antioxidant enzyme superoxide dismutase (SOD) was stimulated, whereas catalase (CAT) and glutathione peroxidase (GPx) were inhibited. Glutathione (GSH) was reduced and glutathione reductase (GR) unchanged. Even though there was an increase in SOD activity, the CAT and GPx did not increase accordingly, maybe due to the direct scavenging of H₂O₂ by salicylcurcumin. The protein content also increased in the curcumin-fed animals, indicating a positive growth-promoting effect. Therefore, it would be beneficial to supplement salicylcurcumin along with the aquaculture feed in order to help the fish to cope with adverse conditions in the environment. This would increase the survival rate, disease resistance and ultimately the growth rate.     

In vivo protective effect of dietary curcumin in fish Anabas testudineus (Bloch)

The present study describes, for the first time, the protective effect of natural curcumin in vivo in a lower vertebrate, a teleost, Anabas testudineus (Bloch). Two doses of curcumin 0.5 and 1% were supplemented in the 40% protein feed and fed to fish for the periods, 2 and 8 weeks. The antioxidant status, protein content, and the tissue structure in experimental fish were examined after the short-term and long-term feeding. In all the curcumin fed groups, the lipid peroxidation product, thiobarbituric acid reactive substances content either decreased or unaffected. The glutathione content increased while the antioxidant enzyme activity pattern varied with time and dose. The histological analysis also confirmed the safety of curcumin retaining the normal arrangement of hepatocytes, hepatopancreas, macrophage–melanocyte centers in Anabas. The improved antioxidant status and protein content suggest a favorable effect for curcumin in cultured fish.     

Local endemism within the Western Ghats-sri Lanka biodiversity hotspot

The apparent biotic affinities between the mainland and the island in the Western Ghats-Sri Lanka biodiversity hotspot have been interpreted as the result of frequent migrations during recent periods of low sea level. We show, using molecular phylogenies of two invertebrate and four vertebrate groups, that biotic interchange between these areas has been much more limited than hitherto assumed. Despite several extended periods of land connection during the past 500,000 years, Sri Lanka has maintained a fauna that is largely distinct from that of the Indian mainland. Future conservation programs for the subcontinent should take into account such patterns of local endemism at the finest scale at which they may occur.