Estimation of micro-flora associated with different stages of Macrobrachium rosenbergii (de Man)

The bacterial flora occurring in muscle, haemolymph, hepatopancreas and gill of brood, juveniles, water, eggs, larvae and rearing water were estimated by selective plate count technique for Entrobacteriaceae, Streptococaceae and Vibrionaceae members. The total viable bacterial count was estimated by total plate count technique on nutrient agar. The total viable counts of bacteria were lowest in water from 6.10x10² CFU/mL) and highest in egg (6.06x10super(8) CFU/g). In brood the total counts were varying from 1.62x10² CFU/g in muscle to 2.20x10super(5) CFU/g in gills. In juveniles, the total plate counts were varying from 2.8x10super(4) CFU/g in muscles to 3.67x10 super(8) CFU/g in hepatopancreas. Selective plate counts show that Enterobacteriaceae members dominate in egg and gills of brood and hepatopancreas of juveniles. Vibrios were found to be dominant in water and larvae of rearing tank. Haemolymph of brood was sterile and did not contain any bacteria while muscle of juvenile was having very low count of total viable bacteria.

Fish-makhana (Euryale salisb) integration: a case study of sustainable aquafarming system in north Bihar

The paper deals with a technique to synchronize two crops, fish and makhana (Euryale ferox Salisb) in a pond. In such eco-friendly integration both crops are mutually benefited. Decomposed plant parts of makhana crop form organic matter that releases nutrients in the water to enhance plankton population. Organic detritus not only acts as food for bottom dwelling fishes (mrigal and common carp) but also provides a suitable substratum for the growth of zooplankton, insect larvae, nematodes and gastropods. Fishes contribute to the control of makhana pests. Their faecal matter acts as organic manure for makhana crop. Plankton population fluctuated between 1260 u/l to 4030 u/l in the control pond and 1630 u/l to 4722 u/l in the experimental pond. During the grand growth period of makhana crop (April to July) the dissolved oxygen content fluctuated between 5.02 mg/l to 6.68 mg/l in the covered areas and 6.04 mg/l to 6.92 mg/l in uncovered areas. Makhana leaves acting as blanket barrier over the water surface brought down the D.O. content in the covered areas of the pond. Free CO sub(2) content showed wider fluctuation in the experimental pond (25.2 mg/l to 30.9 mg/l) than in the control pond (25.1 mg/l to 28.6 mg/l). This could be due to decomposition of plant parts of the presiding crop lying as debris at the pond bottom. Autochthonous supply of nutrients enhanced the content of nitrogen, phosphorous and organic carbon in the soil of experimental pond. The experimental pond covering an area of 0.40 ha yielded 852 kg fish and 200 kg pops whereas the control pond covering the same area produced 777 kg fish only. The net profit per ha came out to be Rs.1,04,700 and Rs. 66,200 in integrated and non-integrated system respectively. Owing to crop diversification, the present integrated system was found to be more viable than the non-integrated system in terms of production and net profit.

The bacteriology of oil sardine (Sardinella longiceps) and mackerel (Rastrelliger kanagurta) caught from tropical waters off Cochin. 1 - Quantitative aspects

The total aerobic viable plate counts (TPCs) of skin, gills and intestine of newly caught oil sardine (Sardinella longiceps) and Indian mackerel ( Rastrelliger kanagurta) at four different temperatures, namely 36 ± 1°C, 28 ± 2°C (RT), 8 ± 1°C and 1 ±1°C, are reported. The total plate count at RT of the skin of oil sardine and Indian mackerel were in the range of l0 super(3) to 10 super(7) and 10 super(4) to 10 super(6) per cm², that of gills in the range of 10 super(5) to 10 super(9) and 10 super(4) to 10 super(8) per g and that intestine in the range of 10 super(5) to 10 sueper(9) and 10 super(5) to 10 super(8) per g respectively. The TPCs were markedly affected by the incubation temperature. Incubation at 28 ± 2°C gave the highest count; at 36 ± 1°C and 8 ± 1°C, the counts decreased by nearly 1-2 log cycles from that at RT. Incubation at 1 ± 1°C registered the lowest count. The peak values for bacterial counts of these fishes occurred at different periods of the year.

Effect of incubation period on plate count of raw ices and frozen fish

The total viable counts were estimated in one hundred and sixty five samples of raw, iced and frozen fish using incubation periods of 24, 48, 72 and 96h. For raw fish, 24h and for iced and frozen fish 48h incubation of the plates were found to be adequate. Variation between samples was significant at 1% level for raw iced and frozen samples.

A bacteriological study of the natural flora of edible oyster, Crassostrea madrasensis

The total viable bacterial populations in the oysters and the sea water from the edible oyster farm at Tuticorin were in the range of 10 super(3) to 10 super(4) per ml and 1 super(2) to 10 super(3) per ml respectively. The maximum most probable number of faecal coliform recorded during the one year period of study of both the oysters and seawater were 33 per 100 ml. Pathogenic bacteria (Salmonella sp., Vibrio cholerae, coagulase positive staphylococci and faecal streptococci were absent in oysters and farm water. Study of 197 (98 taken from oyster liquid and 99 from oyster farm water) randomly isolated cultures indicated that gram negative asporogenus rod-like bacteria of the Vibrio, Flavobacterium, Achromobacter and Pseudomonas groups were the dominant flora of the oyster liquid as well as seawater.

A quantitative study of Vibrio parahaemolyticus (Sakazaki et al) in Etroplus suratensis (Bloch) and Metapenaeus dobsoni (Miers) from Cochin backwater

Seasonal variation of Vibrio parahaemolyticus in fish (Etroplus sauratensis) and prawn (Metapenaeus dobsoni) was monitored from March 1982 to February 1983. Analyses of total viable count, vibrio-like organisms, V. parahaemolyticus like organisms and V. parahaemolyticus showed that they occur more in prawn than in fish. In a more polluted environment, the counts of V. parahaemolyticus associated with fish were found to be higher than in prawn.

Notes on the induced maturation and spawning in four-month-old Penaeus monodon Fabricius by eyestalk ablation

The ablation technique consisted of making an incision across the eyeball to allow free flow of fluids while holding the prawn under water, squeezing the eyeball contents outwards, and pinching hard the eyestalk tissue. The cut area heals completely in about a week; no application of antibiotics is necessary. Spent spawners were tagged with thin brass rings (Rodriguez, 1976) around the unablated eyestalk for a separate experiment on rematuration. Two spawning yielding approximately 277,000 eggs were obtained three weeks after ablation, followed four days later by two more spawnings with 160,000 eggs; all four spawners weighed more than 100 g. With a hatching rate of 98% and 78% for the first and second batch, respectively, the spawnings produced viable nauplii. Water temperatures as low as 23 degree C due to a delayed cold spell in March depressed molting; weakened larvae had to be discharged at the mysis stage. Although ovarian development continued, no further spawnings were obtained due mainly to the onset of bacterial and fungal disease. Infection is initiated in injured portions of the exoskeleton, sometimes penetrating right through the muscles to the ovarian tissues. The non-flowthrough conditions and mussel meat feeding led to fouling of the culture water resulting in consecutive mortalities caused by disease. Female P.monodon held in maturation pens were ablated at the age of 15 months (Santiago, et al., 1976); they averaged only 16 g body weight after four months growth in ponds. In another experiment, pond-reared P.monodon females ranging from 50 to 80 g were ablated at approximately seven months (Aquacop, 1977). The present results show a minimum age of four months from postlarve that P.monodon is capable of ovarian development and spawning upon ablation. However, maturation is probably affected by size as well as age - the four-month old females weighed an average of 100 g in contrast to the smaller animals in the earlier experiments.

Notes on ovarian rematuration of ablated sugpo (prawn) Penaeus monodon Fabricius

To what extent spent P. monodon females can remature and spawn successive broods is an important question in terms of recycling spawners in a commercially viable operation. Corollary to this is the quantity and quality of fry from rematured females in comparison to those from first spawning. Of 347 experimental females, only 10.1% had a second spawning, and 1.4% a third spawning. To a large degree the low rate of rematuration is due to high spawner mortality - average survival period after spawning was only 6 days in a sample of 176 spawners. It took an average of 23 days after ablation for a prawn with undeveloped ovaries to mature and spawn. An ablated female may have another spawning in as little as 5 days after the previous one. Average fecundity was 180,000 eggs per second spawning, and 140,000 eggs per third spawning. The average number of eggs from first spawning ablated females was 110-120,000. Hatching rate was lower for rematuration: 44% for second spawnings, and 35% for third spawnings, as compared to 64% for first maturation.

Inactivation of Vibrio parahaemolyticus in drinking water

A study was conducted in the Cochin area of India to determine the effect of drinking water on Vibrio parahaemolyticus, a bacterium that contaminates fish harvested from marine and estuarine environments. Times of fresh-water exposure required to inactivate these bacteria are given. Findings indicate that the washing of fish and equipment used to handle the fish in drinking water may decrease in the number of viable Vibrio cells and thus aid in prevention of food poisoning.

Effects of different types of feeds on growth and production of tiger shrimp, Penaeus monodon at Bagerhat region, Bangladesh

An experiment was carried out in farmers' gher (shrimp farm) at Bagerhat sadar upazilla, Bagerhat to ascertain the effects of three different types of feeds on the production and economics of brackishwater shrimp, Penaeus monodon for a period of 120 days. There were three treatments such as T1 (BFRI dough feed containing of 30% fish meal, 10% protein conc., 10% soya meal, 15% mustard oil cake, 18% rice bran, 5% maize, 10% wheat flour, 1% oyster shell powder and 1% vitamin premix), T2 (Commercial diet Saudi-Bangla grower) and T3 (Saudi-Bangla special feed). Each treatment had two replicates and the stocking of shrimp in each gher was 3 nos/m². Water quality parameters did not differ significantly among the treatments except water depth. Average production and net return of shrimp in different treatments varied from 404.0 to 509.0 kg/ha and Tk. 56,493.99-Tk. 84,209.60, respectively. T2 showed significantly (p<0.05) the highest production and economic return. The result of the study implied that T2 is more suitable and economically viable than that of other treatments for shrimp farming.

Effects of gamma radiation and -20°C temperatures on the shelf life of Hilsa, Tenualosa ilisha (ham.-Buch. 1822)

The combined effect of radiation and refrigeration on the shelf life of hilsa, Tanualosa ilisha was studied by monitoring the microbiological, chemical and sensory changes of unirradiated and irradiated fish samples using low dose irradiation, doses of 300 krad, 600 krad and 900 krad. Irradiation (900 krad) dramatically reduced population of bacteria, namely total viable counts 48.850cfu per gm for unirradiated, 31.850cfu per gm and 19.600cfu per gm of 300 krad and 600 krad, respectively. The effect was more pronounced at the higher dose (900 krad), total viable count were 14.100cfu per gm. Another microbial indicator total mould counts (TMC) was 8.750cfu per gm, 6.350cfu per gm, and 19.600cfu per gm for 300 krad and 600 krad, respectively. The effect was more pronounced at the higher dose (900 krad) where total viable counts were 14,100cfu per gm. Total volatile nitrogen values increased slowly attaining a value of 101.02mgN per 100gm for unirradiated T. ilisha during refrigerated storage, whereas for irradiated fish, lower values of 71.13, 59.33 and 47.03mgN per 100gm muscle were recorded. Sensory evaluation showed a good correlation with bacterial populations on the basis of overall acceptability scores.

Cloning and expression of zebra fish ferroportin and investigating its influence on anemia

Iron is required for many microbes and pathogens for their survival and proliferation including Leishmania which cause leishmaniasis. Leishmaniasis is an increasingly serious infectious disease with a wide spectrum of clinical manifestations. These range from localized cutaneous leishmaniasis (CL) lesions to a lethal visceral form. Certain strains such as BALB/c mice fail to control L. major infection and develop progressive lesions and systemic disease. These mice are thought to be a model of non-healing forms of the human disease such as kala-azar or diffuse cutaneous leishmaniasis. Progression of disease in BALB/c mice has been associated with the anemia, in last days of their survival, the progressive anemia is considered to be one of the reasons of their death. Ferroportin (Fpn), a key regulator of iron homeostasis is a conserved membrane protein that exports iron across the duodenal enterocytes as well as macrophages and hepatocytes into the blood circulation. Fpn has also critical influence on survival and proliferation of many microorganisms whose growth is dependent upon iron, thus preparation of Fpn is needed to study the role of iron in immune responses and pathogenesis of micoorganisms. To prepare and characterize a recombinant ferroportin, total RNA was extracted from Indian zebrafish duodenum, and used to synthesize cDNA by RT-PCR. PCR product was first cloned in Topo TA vector and then subcloned into the GFP expression vector pEGFP–N1. The final resulted plasmid (pEGFP-ZFpn) was used for expression of FPN-EGFP protein in Hek 293T cells. The expression was confirmed by fluorescence microscopy and flow cytometery. Recombinant Fpn was further characterized by submission of its predicted amino acid sequences to the TMHMM V2.0 prediction server (hidden Markov model), NetOGlyc 3.1 server and NetNGlyc 3.1 server. Data emphasised that obtained Fpn from indian zebrafish contained eight transmembrane domains with N- and C-termini inside the cytoplasm and harboured 78 mucin-type glycosylated amino acid. The results indicate that the prepared and characterized recombinant Fpn protein has no membrane topology difference compared to other Fpn described by other researcher. Our next aim was to deliver recombinant plasmid (pEGFP-ZFpn) to entrocyte cells. However, naked therapeutic genes are rapidly degraded by nucleases, showing poor cellular uptake, nonspecificity to the target cells, and low transfection efficiency. The development of safe and efficient gene carriers is one of the prerequisites for the success of gene therapy. Chitosan and alginate 139 polymers were used for oral gene carrier because of their biodegradability, biocompatibility and their mucoadhesive and permeability-enhancing properties in the gut. Nanoparticles comprising Alginate/Chitosan polymers were prepared by pregel preparation method. The resulting nanoparticles had a loading efficiency of 95% and average size of 188 nm as confirmed by PCS method and SEM images had showed spherical particles. BALB/c mice were divided to three groups. The first and second group were fed with chitosan/alginate nanoparticles containing the pEGFP-ZFpn and pEGFP plasmid, respectively (30 μgr/mice) and the third group (control) didn’t get any nanoparticles. The result showed BALB/c mice infected by L.major, resulted in higher hematocryte and iron level in pEGFP-ZFpn fed mice than that in other groups. Consentration of cytokines determined by ELISA showed lower levels of IL-4 and IL-10 and higher levels of IFN-γ/IL-4 and IFN-γ/IL-10 ratios in pEGFP-ZFpn fed mice than that in other groups. Morover more limited increase of footpad thickness and significant reduction of viable parasites in lymph node was seen in pEGFP-ZFpn fed mice. The results showed the first group exhibited a highr hematocryte and iron compared to the other groups. These data strongly suggests the in vivo administration of chitosan/alginate nanoparticles containing pEGFP-ZFpn suppress Th2 response and may be used to control the leishmaniasis .

Application of whey protein incorporation with Cinnamon oil as a preservative in Huse huso fillet under chilled storage condition

Biodegradable protein-based film was developed by incorporating cinnamon essential oil (CEO) into whey protein concentrate (WPC) at level of 0.8% and 1.5% v/v. Then physical and mechanical properties of the films were evaluated. Adding CEO to the WPC matrix decreased the water vapour permeability of the films and water solubility. Films containing CEO showed significant antibacterial activity both gram-positive and gram-negative strains and exhibited significant inhibitory effect on the studied fungi. In continue, the effect of whey coating and whey coating incorporated with 1.5% CEO on quality and shelf life of Huso huso fillet during refregrated (4±1°C) storage period were also investigated. The control and treated fish samples were analyzed for microbiological (total viable count, psychrophilic counts), chemical (PV, TBA, FFA, pH, TVB-N), and sensory characteristics in 4-day intervals up of microbial, chmical and sensoy analyses indicated lower levels of PV, TBA, FFA, pH, TVB-N in coasted sampels and specially, those with CEO while were kept in refrigerator. Based on results, whey protein edible coating contain 1.5% cinnamon essential oil could enhance preserving ability Huso huso during storage cold.

Effect of thawing methods on the chemical, biochemical, microbiological and sensory indices of Caspian Sea kutum (Rutilus frisii kutum) and rainbow trout (Oncorhynchus mykiss)

Effects of different thawing method i.e. in a refrigerator, in water, at air ambient temperature and in a microwave oven on proximate, chemical (PV, TBA, FFA, TVB-N, SSP, FA), biochemical (pH, WHC,ThL), microbial (total viable, psychrotrophic, coliform, Shewanella and yeast-mould count) and sensory analysis were carried out on frozen whole Caspian sea Kutum (Rutilus frisii kutum) and Rainbow trout (Oncorhynchus mykiss) carcasses. The values of ash, protein, SSP, WHC, PUFA, PUFA/SFA. EPA+DHA/C16:0, pH, and microbial count of thawed samples decreased significantly while fat, PV, TBA, FFA, TVB-N, SFA and MUFA increased compared to the fresh fish (unfrozen) as control samples. Also, sensory evaluation all of thawed samples showed a significant (p<0.05) quality loss compared to the fresh fish as control samples. The lowest chemical and biochemical values as well as microbial growth were determined in water thawed samples. Therefore, based on this study thawing in water is most suitable for frozen whole rainbow trout.

In vivo and in vitro storage of kutum, Rutilus frisii kutum eggs

Effects of post-ovulatory and post-stripping retention time and temperature on egg viability rates were studied in kutum (Rutilus frisii kutum). Eggs were retained inside (in vivo storage) or outside the ovarian cavity with ovarian fluid (in vitro storage) at various temperatures. Two experiments were performed: 1) Partial volumes of eggs were stripped and fertilized at 24- hour intervals for 96 hours post-ovulation (HPO) (at 11 °C) and at 12-hour intervals for 72 HPO (at 14 °C), and 2) stored eggs were fertilized after 0, 2, 4, 6, and 8 hours post-stripping (HPS) at temperatures of 4, 10, 12, and 26 °C. In the first experiment, the highest eyeing and hatching rates (76% and 60% at 11 °C; 81% and 71% at 14 °C) and the lowest eyed-egg mortalities (20% at 11 °C; 12% at 14 °C) occurred in the eggs fertilized immediately (0–24 HPO at 11 °C and 0–12 HPO at 14 °C) after ovulation. Egg viability, as shown by successful eyeing and hatching rates, was completely lost by 72–96 HPO at 11 °C, and 60–72 HPO at 14 °C. In the second experiment, the maximum eyeing (87%) and hatching (75%) rates of eggs took place at 0 HPS followed by 8 HPS (> 80% and > 70%, respectively) at 4 °C. As storage temperature increased, egg viability decreased: 80%, 70%, and 50% viable at 8 HPS at 4, 10, and 12 °C, respectively. The eggs stored at 26 °C lost their viability almost completely after 4 HPS. Eyed-egg mortality increased from 13% at 0 HPS to 48.2% at 4 HPS at 26°C. These results demonstrate that egg stripping should take place within 168 °C-hours after ovulation and that complete loss of viability of the eggs occurs by 672°C-hours after ovulation. The in vivo storage method is more effective compared to in vitro storage. Also successful in vitro storage of eggs can be used atleast within 8 hours at temperatures ranging from 4 to 12ºC.