Ecophysiological response of Nerita oryzarum (Recluz), a gastropod, to variations in temperature, pH and salinity

During ecophysiological investigations on an intertidal gastropod, Nerita oryzarum (Recluz), of Mumbai shore, various biochemical changes could be recorded. Glycogen and lipid contents of N. oryzarum were found to decrease, whereas, water content increased with decreasing salinity. The rate of oxygen consumption declined with the decrease in salinity and also in highly acidic (pH 2) as well as highly alkaline (pH 10) sea water. The observed variations in the rate of oxygen consumption and changes in biochemical composition in the animal with changes in salinity, pH and temperature are probably the process of physiological and biochemical adjustments to the fluctuating environmental conditions in the intertidal region.

In vitro phagocytic study of blood leucocytes and peritoneal macrophages of walking catfish Clarias batrachus ( Ham.) against Aeromonas hydrophila and Escherichia coli.

In observation of in vitro phagocytic activity against Aeromonas hydrophila isolate 34k (a virulent form) and Escherichia coli (an avirulent bacteria) of neutrophil- and monocyte-like cells of walking catfish Clarias batrachus showed phagocytosis. N eutrophils and monocytes phagocytized the avirulent form of bacterial isolate more than the virulent one. Other blood leucocytes did not show phagocytosis. Peritoneal macrophage of the fish were separated by glycogen elicitation and the macrophages were being adhered on plastic cover slips for studying their in vitro phagocytic activity. Most of the cells were alive after adherence and showed phagocytosis against the virulent and avirulent bacteria. The percent phagocytosis and phagocytic index were higher against the avirulent E. coli than the virulent A. hydrophila.

Characterisation of neurons in the pedal ganglia of the green-lipped mussel, Perna canaliculus, using antibodies raised against neuropeptides and neurotransmitters involved in gastropod egg-laying behaviour and bivalve reproduction and spawning

To characterise central neurons in the pedal ganglia of both male and female green lipped mussel, Perna canaliculus immunohistochemical techniques were used. Mollusc antibodies were used against neuropeptides and neurotansmitters known to control reproduction and spawning. Anti-ELH and anti-APGWamide showed very strong immunoreactivity in small type of neurons. Anti-5-HT and anti-DA immunoreactivity was mostly in large type of neurons. The labelled neurons are consistent with descriptions of neurosecretory cells implicated in the control of reproduction and spawning on the basis of earlier histological staining techniques used in this species. The use of selective immunological markers for peptides and amines appears to be a, promising tool for further characterisation of neurosecretory cells, and to isolate an'tl characterise neuropeptides and other biologically active materials involved in the control of reproduction in Perna canaliculus.

Effect of feed quality restriction followed by realimentation on growth, nutrient utilization, biochemical changes and haematological profiles of Indian major carp, rohu (Labeo rohita H.)

To investigate the effect of protein restriction with subsequent re-alimentation on nutrient utilization, hematological and biochemical changes of Indian major carp, Rohu (Labeo rohita H.), 150 acclimatized Rohu fingerlings (average 20.74 ± 0.13 g) divided into five experimental groups (30 fingerlings in each groups with three replications with 10 fingerlings in each) for experimental trial of 90 days using completely randomized design. Control group (T sub(CPR)) was fed with feed having 30% crude protein at 3% of body weight for 90 days trial period. Other experimental groups T sub(1PR) was alternatively 3 days fed with feed having 20% CP and 30% CP at 3% of body weight, T sub(2PR) was alternatively 7 days fed with feed having 20% CP and 30% CP at 3% of body weight, T sub(3PR) was alternatively 15 days fed with feed having 20% CP and 30% CP at 3% of body weight and T sub(4PR) was alternatively 25 days fed with feed having 20% CP and 30% CP at 3% of body weight during 90 days trial period with daily ration in two equal halves at morning and afternoon. It was noticed that retention of different nutrients was almost similar among all treatment groups indicated improvement of digestibility of nutrients might not be the mechanisms for recovery growth in carps. Increased percent feed intake of body weight (hyperphagia) (4.14 ± 0.30 or 4.94 ± 0.46 and 3.33 ± 0.29), improved specific growth rate (1.86 ± 0.09 or 2.26 ± 0.05 and 1.43 ± 0.01), absolute growth rate (1.57 ± 0.08 or 1.84 ± 0.18 and 1.36 ± 0.12), protein efficiency ratio (1.19 ± 0.11 or1.16 ± 0.12 and 1.05 ± 0.09) were the important mechanism showing better performance index (21.60 ± 1.09 or 23.80 ± 0.21 and 19.45 ± 0.37) through which the experimental groups which were protein restricted and re-alimented at 3 or 7 days alternatively during 90 days trial period could able to compensate the growth retardation and to catch up the final body weight of control (128.68 ± 11.53 g/f) but other experimental groups failed to compensate during 90 days trial period. Result of the present study indicated that deprived fish i.e., fish received alternate 3 or 7 days protein restriction and re-alimentation showed recovery growth had still lower values of Hb (10.21 ± 0.02, and 9.88 ± 0.04 g/dl), hematocrit value (30.62 ± 0.05 and 26.64 ± 0.11%), total erythrocytic count (3.40 ± 0.01 and 3.29 ± 0.01 X10super(6) mm³), plasma glucose (126.93 ± 0.20 and 126.67 ± 0.05 mg/dl), total plasma lipid (1.04 ± 0.01 and 1.02 ± 0.01 g/dl) and liver glycogen (290.10 ± 0.80 and 288.99 ± 0.95 mg/kg) in comparison to control (10.56 ± 0.08 g/dl, 31.68 ± 0.24%, 3.52 ± 0.03 X10super(6) mm³, 128.23 ± 0.25 mg/dl, 1.07 ± 0.01g/dl and 292.00 ± 0.23 mg/kg) at the end of 90 days trial but total plasma protein in deprived group was compensated with advancement of trial period. All hematological and biochemical parameters studied were proportionately lowered in the experimental group got higher degree of deprivation. These findings suggested that with the increase of trial length complete compensation of hematological and biochemical profiles of rohu might be achieved. The results indicated that the implementation of alternative 7 days low and high protein diet feeding during aquaculture of carps could make economize the operation through minimizing the feed input cost.

Some biochemical constituents and the calorific values of different regions of the body musculature of pond murrel Channa punctatus Bloch

A study of the distribution of some chemical constituents in the musculature of Channa punctatus Bloch showed it to be fairly heterogeneous. The increase in the levels of protein, fat, ash, total carbohydrates, glycogen, RNA, DNA and cholesterol towards the posterior region of the fillet was the result of increase in the number of cells per unit weight of the sample and in the concentration of myocommata. The distribution of water in the musculature was inversely related to that of the fat. The concentration of protein appeared to be associated with the RNA level. The posterior region of the fillet exhibited more calorific value than the anterior region.

Seasonal changes in the biochemical composition of liver in Garra mullya (Sykes)

Seasonal changes taking place in the biochemical constituents of liver of G. mullya are reported. An inverse relationship was noticed in the variation of fat and water. Maximum fat contents were observed during june-july. Reserve fat was utilized through gluconeogenesis during the spawning months. Protein and glycogen percentages were comparatively higher in liver than in the muscles and gonads. Decline in the glycogen content was associated with spawning during July to November. Nutritive values have shown more energy contents in the liver during pre-spawning months.

Technological aspects of preservation and processing of edible shell fish 2. Influence of season on the chemical composition of crab (Scylla serrata)

The purpose of this communication is to bring out the influence of season on the chemical composition of crab, covering a period of 2 years. Changes in moisture, protein, water extractable nitrogen, non-protein nitrogen, glycogen, lactic acid, fat and free amino acid composition of crab meat have been reported on a monthly basis.

Purification and characterisation of phosphorylase from muscle of Tilapia (Tilapia mosambica)

Phosphorylase from muscle of tilapia (Tilapia mosambica) was extracted by water and purified by ammonium sulphate precipitation, centrifugation and repeated recrystallisation. Electro-phorogram of the enzyme preparation showed a single band near origin. The enzyme showed optimum pH and temperature at 6.1 and 37°C respectively. Glucose and glucose-6-phosphate were found to be competitive inhibitors of the enzyme. Maltose and starch acted as primers for the phosphorylase reaction like glycogen.

Studies on the ice storage characteristics of blood clam Anadara granosa meat

Ice-storage study of blood clam (Anadara granosa) meat in direct contact and out of contact (in 200 gauge polyethylene bag) with ice was taken up to assess the amenability of the meat to icing. Changes in moisture, total protein, non-protein nitrogen, α amino nitrogen, total volatile base nitrogen, glycogen, free fatty acid, peroxide value, total bacterial count and coliform count were followed every day. The raw and cooked meat were also subjected to organoleptic evaluation. The study showed that the clam meat can be ice-stored in very good condition out of contact with ice in polyethylene packets for 4 days and in direct contact with ice for 2 days.

Neurons characterization in the cerebral ganglia of the green-lipped mussel, Perna canaliculus, using antibodies raised against neuropeptides and neurotransmitters involved in gastropod egg-laying behaviour and bivalve reproduction and spawning

Immunohistochemical techniques were used to characterise central neurons in the cerebral ganglia of both male and female Perna canaliculus. We used mollusc antibodies raised against neuropeptides and neurotransmitters known to control reproduction and spawning. Anti-ELH and anti-APGWamide showed very strong immunoreactivity in small type of neurons. Anti-5-HT and anti-DA immunoreactivity was mostly in large type of neurons. The labelled neurons are consistent with descriptions of neurosecretory cells implicated in the control of reproduction and spawning on the basis of earlier histological staining techniques used in this species. The use of selective immunological markers for peptides and amines appears to be a promising tool for further characterisation of neurosecretory cells, and to isolate and characterise neuropeptides and other biologically active materials involved in the control of reproduction in Perna canaliculus.

Optimization of dose of methyltestosterone (MT) hormone for sex reversal in tilapia (Oreochromis niloticus L.)

This paper describes the optimization of dose of methyltestosteronei (MT) hormone for masculinization of tilapia (Oreochromis niloticus). Five treatments (i.e. T1 T2, T2, T4 and T5) with different doses such as 0, 40, 50, 60 and 65 mg of MT hormone were mixed with per kg of feed for each treatment and fed the fry four times a day up to satiation for a period of 30 days. The stocking density was maintained 10 spawn/liter of water. The growth of fry at different treatments was recorded weekly and mortality was recorded daily. At the end of hormone feeding the fry were reared in hapas fixed in ponds for another 70 days and at the 100th day the fish were sexed by the gonad squashing and aceto-carmine staining method. The analysis of growth data did not show any significant variation in length and weight of fish among the different treatments. High mortality of fry ranging 66% to 81.6% was observed in different treatments and highest mortality was observed during the first twelve days of the experiment. The sex ratio analysis showed that T2 (40 mg/kg) and T5 (65 mg/kg) produced 93.33% of sex reversed male and T3 (50 mg/kg) and T4 (60 mg/kg) produced 96.66% sex reversed male, and these ratios were significantly (p<0.05) different from 1:1 male: female sex ratio. The control, T1 (0 mg/kg) contained 43.33% male progeny. From these results it is suggested that either 50 mg/kg or 60 mg/kg of MT with a feeding period of 30 days could be considered as an optimum dose for masculinization of tilapia (O. niloticus).

Genetic diversity study of Caspian brown trout (Salmo trutta caspius Kessler, 1877) population in five rivers in the southern part of Caspian Sea, Iran

For study the genetic diversity of Caspian brown trout population in five rivers in the southern part of Caspian Sea in Iran 182 number generators in the fall and winter of 1390 were collected in Chalus, Sardab Rud, Cheshmeh Kileh, Kargan Rud and Astara rivers. Then about 3-5 g of soft and fresh tissue from the bottom fin fish removed and were fixed in ethanol 96°. Genomic DNA was extracted by using ammonium acetate, then quantity and quality of the extracted DNA were determined by using spectrophotometry and horizontal electrophoresis in 1% agarose gel. The polymerase chain reaction was performed by using 16 SSR primers and sequencing primers (D-Loop) and the quality of PCR products amplified by SSR method were performed by using horizontal electrophoresis in 2% agarose gel. Alleles and their sizes were determined by using vertical electrophoresis in 6% polyacrylamide gel and silver nitrate staining method. Gel images were recorded by gel documentarian, the bands were scored by using Photo- Capt software and statistical analysis was performed by using Gene Alex and Pop Gene software. Also the PCR sequencing products after quality assessment by usinghorizontal electrophoresis in 1.5% agarose gel were purified and sent to South Korea Bioneer Corporation for sequencing. Sequencing was performed by chain termination method and the statistical analysis was performed by using Bio- Edit, Mega, Arlequin and DNA SP software. The SSR method, 5 pairs of primers produced polymorphic bands and the average real and effective number of alleles were calculated 5.60±1.83 and 3.87±1.46 in the Cheshmeh Kileh river and 7.60±1.75 and 5.48±1.32 in the Karganrud river and the mean observed and expected heterozygosity were calculated 0.44 ±0.15 and 0.52 ±0.16 in the Cheshmeh Kileh river and 0.50 ±0.11 and 0.70±0.13 in the Karganrud river. Analysis of Molecular Variance results showed that significant differences in genetic diversity between and within populations and between and within individuals in the studied rivers (P<0.01). The sequencing method identified 35 different haplotype, the highest number of polymorphic position (251) and haplotype (14) were observed in the Chalus river. The highest mean observed number of alleles (2.24±0.48) was calculated in the Sardabrud river, the highest mean observed heterozygosity (1.00±0.03) was calculated in the Chalus river and the highest mean nucleotide diversity (0.13±0.07) was observed in the Sardabrud river and mean haplotype diversity was obtained (1) in three studied rivers. The overall results show that there are no same population of this fish in the studied rivers and Karganrud and Chalus rivers in the SSR and sequencing methods had the highest levels of genetic diversity.

Survey of biosystematic and sexual reproductive physiology in Holothuroidea on Hormozgan Province

Sea cucumbers belong to phylum Echinodermata, order Holothuroidea are an abundant and diverse group of Invertebrates, with over 1400 species occuring from the intertidal to the deepest oceanic trenches. Sea cucumbers are important components of the food chain in temperate and coral reef ecosystems and they play an important role as deposite feeders and suspension feeders. Rapid decline in populations may have serious consequences for the survival of other species that are part of the same complex food web,as the eggs, larve and juveniles constitute an important food source for the other marine species including crustaceans, fish and mollusks. In addition sea cucumbers are often called the earthworms of the sea, because they are responsible for the extensive shifting and mixing of the substrate, and recycling of detrital matter. Sea cucumbers consume and grind sediment and organic material into finer particles , turning over the top layers of sediment in lagoons , reefs and other habitats and allowing the penetration of oxygen. While the taxonomy of the holothurian families is generally well known , the distinction of similar species is difficult. There are relatively few holothurian taxonomist.Most sea cucumber species can be identified by Holothurin taxonomists by using the calcareous skeletal ossicles found in the body wall. In this study , at first a sea cucumber from Kish island in Persian gulf has recognized. Individuals collected from west and east extend far away into north and south of coral reefs by diving. I have checked them morphologically and anatomically.Then with key to the orders of the Holothuroidea, They belong to the Aspidochirotida with key to the families of Aspidochirotida, they were in Stichopodidae families and with key to the genus of Stchopodidae, they were Stichopus. Then ossicles were extracted at National Museum of Natural History, by Dr David Pawson. The ossicles were measured on a transect across a slide prepared from the mid-dorsal region of each specimen.The one we have in the shallow waters of Kish island, is Stichopus hermanni, a massive holothurian, body broad, considerably flattened ventraly ,the dorsal side slightly arched and the lateral sides almost vertical; body wall fairy thick and soft ; mouth subterminal; anus central; tentacles usually 20 in number of length and leaf shaped. Numerous ossicles consisting of table with large discs having usually 7 to 15 peripheral holes, but often irregular or incomplete and spire of moderate height ending in a group of spinelets, rosettes of variable development, and c-shaped rods. Color (exept papillae)partly remained after preservation in alcohol which is found at the depth of 4 to 8 meters, on coral reef. Furthermore, the sexual reproductive cycle was described using standard methods. Gonads were removed and transferred to Bouin's fixative for four weeks and then processed according to standard embedding technique. To prevent the loss of tubule contents during embedding, the tubule sections, were cut well beyond the segment selected for sectioning. For each individual, six sections, each section with 5µm diameter by microtome were cut from tubules. These sections were first placed on gelatin coated slides (the gelatin was heated to 42°c) and then transferred to the oven at 37°c for one hour. This technique usually prevents the fragil tubules from breaking and the loss of gametes. The slides were stained with Eosin and Hematoxylin, and good resolution of the various cell types achieved.A second series of slides was stained with the Periodic Acid Schiff(PAS) to identify polysaccharides(glycogen). Monthly sampling was occurred.The sexual reproductive cycle was defined through the combined use of these criteria: Monthly percentages of the gonad stages for each sex, the monthly gonad index (GI) , given as the ratio of the wet gonad weight (G) to the dray weight (DW)and the monthly percentage of individuals that undetermined sex. The gonad consists of two tufts of tubules on which saccules develop. Gonadal development was classified into five stages: post spawning, recovery, growth, advanced growth, and mature stage that were adapted from the earlier studies of holothurians. Histological preparations showed that the sex of larger individuals could be identified by the presence of oogonia and young oocytes in females, and spermatogonic stages in males.The mean diameter of the tubules and gonadal mass follow annual cycles, increasing from late winter through spring, and dropping abruptly after spawning in the summer. Gametogenesis is generally a prolongate process and begins in March. By summer the ovarian tubules contain oocytes with diameter of 120-240 pm and the testicular tubules contain an abundance of spermatozoa (diameter 5-6 gm ).Following spawning the predominant activity within the spent tubules is phagocytosis of the residual gamets.The active phase of gametogenesis (March to July), coincides with an increasing photoperiod regim, and an accelerated gametogenesis occurs in July when temperature is high. Throughout the year, the gonad of Stichopus hermanni is larger in males than in females, and this is due to the number of tubules in the testis rather than to tubules length or diameter.