Maximins S, a novel group of antimicrobial peptides from toad Bombina maxima

Amphibian skin secretions are rich in antimicrobial peptides acting as important components of innate defense system against invading microorganisms. A novel type of peptide, designated as maximin S, was deduced by random sequencing of 793 clones from a constructed Bombina maxima skin cDNA library. The putative primary structures of maximin S peptides can be grouped into five species, in which maximin S I has 14 amino acid residues and the rest of maximin S peptides (S2-S5) all have 18 amino acid residues. Unlike most of the amphibian antimicrobial peptides so far identified, the newly characterized four maximin S precursors are composed of maximin S I and different combinations of tandem repeated maximin S2-S5 linked by internal peptides. Except maximin S I, the predicted secondary structures of maximin S2-S5 show a similar amphipathic alpha-helical structure. MALDI-TOF mass spectrometry analysis of partially isolated skin secretions of the toad indicates that most of the deduced maximin S peptides are expressed. Two deduced maximin S peptides (S1, S4) were synthesized and their antimicrobial activities were tested. Maximin S4 only had an antibiotic activity against mycoplasma and had no antibacterial or antifungal activity toward tested strains. Maximin S1 had no activity under the same conditions. (C) 2004 Elsevier Inc. All rights reserved.

Effect of modified starch on the shelf life of frozen fish Kamaboko with prepared spinach

Fish kamaboko with spinach was prepared by mixing fish kamaboko with spinach prepared with different combinations of ordinary starch (OS) and modified starch (MS), viz., 40:60, 50:50, 60:40, 100%) MS and 100% OS. The products were frozen at -40°C and stored at -20°C, and subjected to biochemical (peroxide value, total titratable acidity, pH and moisture) organoleptic (appearance, taste and colour), microbiological (total plate count) and physical (folding and expressible water) tests at monthly intervals. Among the different combinations tried, it was observed that fish kamaboko with spinach prepared with 50:50 and 60:40 combinations of OS and MS could be stored for six months at-20°C.

Morphological overlap of fish community in Vamanapuram River, south Kerala

The morphological characteristics and overlap of fish community in Vamanapuram River have been studied in detail. In the 12 study sites, 19 fish species were encountered. Based on the body shape, four different types are apparent. The elongate bodied fishes (RBD<1.5) like Hemiramphus xanthopterus and Xenentodon cancila are grouped under one category. The deep bodied fishes (RBD>3.5) like Puntius filamentosus, P. ticto, P. vittatus, P. melanampyx, P. sarana, Etroplus maculatus and E. suratensis come under a separate category. Fishes with round to square cross section like Garra mullya and Glossogobius giuris form a separate group. All the other species are grouped as generalized bodied fishes. The morphological overlap studied for the Vamanapuram fish community showed that out of 190 combinations, 30 combinations have high overlaps (≥ 67). P. melanampyx has maximum number (6) of high overlaps. Puntius spp., which constituted 49.5% of the total population, have a mean morphological overlap of 52%. The morphological overlap of fish species in relation to the trophic structure is discussed in detail.

Standardisation of recipe for the preparation of lobster analogues

Lobster analogues were prepared with lobster base flavour (paste),h lobster cook water (whole lobster omogenate:water, 1:1 and 1:0.5) and lobster meat mince. In another experiment, different combinations of ginger-garlic paste and lobster base flavour, i.e., 1:3, 1:4, 1:6, 3:3, 3:4, 3:6, 5:3, 5:4 and 5:6 were added to the lobster analogue paste. It was observed that lobster analogues prepared with lobster base flavour (paste) are suitable organoleptically. The combination of ginger garlic paste and lobster base flavour in the ratio of 3:4 was found to be suitable organoleptically. Lobster analogues coloured with annatto seed colour at 1:2 (annatto seed:water) concentration had high values for the colour attribute as compared to orange-red synthetic colour, beetroot colour, caramel colour and paprika colour. It was observed that come-up-time to achieve a temperature of850°C was 28 minutes with a processing period of 11 minutes.

Seed production and culture techniques of Genetically Improved Farmed Tilapia (GIFT) in brackishwater environment

Effects of different levels of salinity on survival, growth and gonadal development of Genetically Improved Farmed Tilapia (GIFT) were studied under laboratory conditions in glass aquarium, for a period of ten weeks. The initial individual size of the GIFT was 20.23±4.45 and the salinity levels tested were 0, 5, 10, 15 and 20 ppt. The highest survival of 87.5% was found in 0 ppt and the lowest 60.5% in 20 ppt. Though the survival decreased progressively with increased salinity, there were no significant differences (P>0.05) among 0, 5, and 10 ppt. Similar to what has been observed in survival, the specific growth rate (SGR %/day) also decreased as of 1.30, 1.24, 1.08, 0.90 and 0.71, respectively, with the increased salinity of 0, 5, 10, 15 and 20 ppt. The gonadal development was highest in 0 ppt with a GSI value of 3.75 and lowest of 2.01 in 20 ppt. In the second experiment, gonadal development and seed production performance of GIFT in brackishwater condition were investigated for a period of three months. Each of the three fine meshed hapas of 20 square meters made from nylon net was placed in a freshwater (0 ppt) and in a brackish water (10-15 ppt) pond of the Brackishwater Station (BS). GIFT of 65 g average weight from a single cohort were stocked into three hapas at a rate of 2 per m. The male vs female ratio was 1:3. The development of gonad was faster with the higher gonadosomatic index (GSI %) of 3.85 % in freshwater condition than that of 2.73 % in brackish water. Within three months of the study period, a total of 70,510 and 44,250 GIFT fry were produced respectively, in freshwater and brackishwater conditions. Finally under third experiment, a participatory on-farm trial was carried out to evaluate the production performance of GIFT in monoculture and in polyculture with silver barb in coastal freshwater pond conditions. Nine ponds were selected for three treatment combinations of GIFT monoculture (T1), GIFT and silver barb polyculture (T2), and silver barb monoculture (T3). The ponds have been stocked in April, 05 at a density of 25,000 fry per ha. Fishes were fed with rice bran at the rate of 6% bw per day. In one month culture period, GIFT attained an average weight of 16.27 g in monoculture and 17.23 g in polyculture, against an average stocking weight of 0.37 g. Silver barb reached an average weight of 16.62 g in polyculture with GIFT and 10.01 g in monoculture, against an average stocking weight of 3.79 g.

Studies on the chemical control of psychrophilic bacterial spoilage of fish. 2 - The effect of antibiotics on the growth of psycrophilic bacteria isolated from marine fish

Among the various antibiotics tried, tetracyclines particularly chlorotetracycline (CTC), chloramphenicol and chlorostrep were found to be fairly effective at 8 and 10 p.p.m. levels. The order of sensitivity to CTC among the six genera studied was found to be Achromobactercombinations of antibiotics for commercial application are discussed.

Selection of suitable diluents for bacteriological examination of fishery products

For raw, iced and frozen samples of fish and prawn, significant difference was observed in total plate counts done with various diluents, the significance level ranging from 5% to 0.1%. For raw fish, N-saline, seawater and quarter strength Ringers' solution gave maximum total plate counts. In the case of iced-fish, n-saline yielded highest total plate counts. For frozen samples, however, peptone water and n-saline gave good recoveries. Trials with suitable combinations of diluents showed that though some of them were as good as the control, namely n-saline, none were superior in any way.

A demersal trawling gear for inshore waters along the west coast

Results of comprehensive efficiency tests of three tested designs of 15m bulged belly, 15.8m six seam and 29.26m longwing type trawls in combination with 114x57cm both rectangular flat and horizontal curved wooden otter boards are reported. Of the possible six combinations, the bulged-belly trawl with flat rectangular otter boards has performed better in landing prawns. Further this study has indicated the selective action of the different designs.

Phantom mutation hotspots in human mitochondrial DNA

Phantom mutations are systematic artifacts generated in the course of the sequencing process. Contra common belief these artificial mutations are nearly ubiquitous in sequencing results, albeit at frequencies that may vary dramatically. The amount of artifacts depends not only on the sort of automated sequencer and sequencing chemistry employed, but also on other lab-specific factors. An experimental study executed on four samples under various combinations of sequencing conditions revealed a number of phantom mutations occurring at the same sites of mitochondrial DNA (mtDNA) repeatedly. To confirm these and identify further hotspots for artifacts, > 5000 mtDNA electropherograms were screened for artificial patterns. Further, > 30000 published hypervariable segment 1 sequences were compared at potential hotspots for phantom mutations, especially for variation at positions 16085 and 16197. Resequencing of several samples confirmed the artificial nature of these and other polymorphisms in the original publications. Single-strand sequencing, as typically executed in medical and anthropological studies, is thus highly vulnerable to this kind of artifacts. In particular, phantom mutation hotspots could easily lead to misidentification of somatic mutations and to misinterpretations in all kinds of clinical mtDNA studies.

Occurrence of enterotoxigenic staphylococci in frozen fishery products

Sixty six samples of frozen fishery products consisting of frozen crab meat, frozen prawns, frozen cuttlefish and frozen mussel collected from nine cold stores situated in and around Cochin city were examined for coagulase positive staphylococci. Forty eight samples (72.72%) revealed the presence of coagulase positive staphylococci, which were then tested for enterotoxigenity, employing cellophane-over-agar method for toxin production and optimum-sensitivity-plate (OSP) method for toxin detection. Seventy seven out of one hundred and two (75.49%) coagulase positive staphylococcal isolates from the above products produced enterotoxins A, B, C, D and E either singly or in combinations. Enterotoxin A and D were detected more often than others. The occurrence of enterotoxigenic staphylococci in frozen fishery product is reported for the first time in this country.

Preliminary chemical and physical evaluation of some formulated feeds for P. monodon

The culture of Penaeus monodon has explicitly defined the need for diet formulations or supplementary feeds that would promote optimum growth and survival of the animal. A total of 28 feed combinations were developed for P. monodon. Fish meal, shrimp head meal, squid head meal, Ascetes spp. rice bran, and soybean cake were used as primary ingredients in these feeds. The commercial vitamin mix No. 22 was added to the dry ingredients. Gelatinized corn starch and wheat flour were used as binders. The pellets were extruded using a portable kitchen grinder with a diameter of 4 mm. The products were either sun-dried for 8 hours or oven-dried overnight at 50 degree C to stabilize moisture at 8-10%. The pellets were then kept in covered glass bottles and stored in the laboratory at room temperature. The cost of the feeds excluding labour were also computed. The pellets were analyzed for protein, fat, carbohydrate, crude fiber, ash, and moisture contents using standard procedures. They were also analyzed for water stability. To test the stability of pellets in water, 2-g samples were placed in plankton nets (mesh #40) and suspended in water for two, and six hours. The undissolved samples were then vacuum-dried and the moisture determined. Cost of the feeds ranged from P1.10 to P2.60 per kg depending on the feed ingredient. Squid and Ascetes spp. were rather expensive for use as basic ingredients. Proximate analysis of dry weight showed percentage protein content ranged from 20-63 g; fat, 8-20 g; carbohydrate (by difference), 11-36 g; ash, 8-28 g; moisture, 6-11 g; and crude fiber, 5 . 13 g. Stability tests showed that after two hours, 35-88% of solids remained intact and after 6 hours, 20-55% of the pellets remained undissolved. When a pellet disintegrates easily, pollution of the water occurs. Chances for the shrimp to feed on the pellet is minimized when the pellet is unstable. Thus, the search for a more compact feed pellet has to be continued.

Evaluation of milkfish (Chanos chanos Forskal) and prawn (Penaeus monodon Fabricius) in polyculture systems

The growth, survival and profitability of C. chanos and P. monodon grown in 5 different combinations for 100 days in 500m2 brackish water ponds were assessed. Differences in the growth and production of prawns cultured singly or in combination with milkfish at increasing stocking density strongly suggests that the presence of milkfish exerts some negative effect on prawn. However, growth production and competition index data suggest that the presence of prawn do not significantly affect milkfish. While the maximum production of prawn can be attained in monoculture, its polyculture with 2000 milkfish/ha is also economically feasible.

The study of maturational progress in the whitespotted rabbitfish (Siganus sutor) under different photoperiod and light intensity conditions

The rule of light on the timing of maturation and spawning in tropical and subtropical regions is not clear well, because the reproductive cycle in these systems is lunar synchronized. In this study, thus, the effects of different light regimes were investigated on maturational progress of whitespotted rabbitfish, Siganus sutor, the commercial species in Persian Gulf and Oman Sea. During prespawning season, 50 adult fish were randomly divided into ten 300-l tanks (n=5). The fish in control tank received indoor light condition and the fish in each other tanks were exposed to nine different combinations of photoperiod (8L: 16D, 12L: 12D, 16L: 8D) and light intensity (1000, 2000, 3000 lux). After 60 days, GSI and HSI values, serum levels of estradiol (E2), testosterone (T), 17-α hydroxyprogestrone (OHP), calcium (Ca2+) and gonad histology were evaluated for females and males. In females the GSI mean values of exposed fish increased in comparison with control except for fish were kept under 8L, 2000 lux (tank 8). These differences were significant only for fishes in tank 7 (8L, 3000 lux). In the cases of HSI, the results were converse, so that, the most of thanks showed significant decreasing than fishes reared under indoor condition. Morphology and histology study of Ovaries showed three developmental phases including 3, 4 and chiefly 5 that were parallel with GSI values. Fortheremore the serum levels of E2 was recorded between 0.54-15.8 ng/ml in different fish and their mean values were lower than control in all treatmants (P> 0.05). In males, the similar results were obtained. The GSI and HSI mean values in experimental regimes compared with control were upper and lower, respectively, except for fishes were reared in tank 1 (16L, 3000 lux). Testes histology of fishes were reared under different regimes showed signs of stage 5, since no blood vessels observed and thick milt exuded on slight pressure. The mean values of testosterone consentration in fishes were kept in tanks 1 and 6 (12L, 1000 lux) were higher and in other ones were lower than control group. It is also noted that the OHP and Ca2+ had diverse results including increasing and decreasing mean values than control. So, these factors contrary to E2 and T were not suitable to evaluation of maturity in both sexes. On the basis of ovarian structure in stage 5, oocyte development pattern in this species was group synchronous. So, increased mean of GSI versus decreased values of HSI, E2 and perhaps Ca2+ were the signs of improved maturation. But in males, reduced levels of T and similarity of testes morphology in all samples caused that GSI mean value was the only indicator for differentiating among treatments. These findings suggest that alternations were used in light regimes have been the reason of improved maturity in all treatments except fishes reared in tank 8. The ١٠٧ rule of light intensity on induction of maturity was cleared by comparision between fishes in tanks 4, 5, 6 and control group. Because day length was the same whereas fishes in tanks 4, 5 and 6 were exposed to increased light intensity compared with control. This fact verified by results was obtained from fishes in tanks 9 and especially 7, since photoperiod in this group was lower than control. So, higher intensity was considered as the reason of alternations. Contrasting with indoor condition, Induced maturity was also cleared for fishes were kept in tanks 1, 2 and 3, where both of light duration and intensity were increased. But, the rule of photoperiod was individually demonstrated when obtained results were compared with similar light intensities in other treatments. In conclusion, with comparison among different light regimes it is declared that siganids were kept under light condition of tank 2 including 16h light duration combined with 2000 lux intensity showed the best signs of sexual maturation in both sexes. On the basis of this study, setting up the spring light condition during prespawning season induces maturation in withspotted rabbitfish. This improvement not only by influence of photoperiod or light intensity, separately, but obtained through interaction between them. Thus, determination of threshold and resistance to light be recommended before exposure, although using proper regime during suitable time are necessary to achieve purposes considerably.

Genetic diversity of Bartail flathead (Platycephalus indicus (Linnaeus,1758)) using AFLP markers in Persian Gulf

Platycephalus indicus is a large benthic fish that inhabits temperate and tropical coastal waters of the Indo-West Pacific and found on sand or mud bottom in vary shallow area of estuary and near shore to depth of 25m. This species is dominant species of platycephalidae family, in Khuzestan, Bushehr and Hormozgan provinces and mainly is captured by bottom trawl, gillnet and moshta in Hormozgan. This study was designed to evaluate population variation and differentiation of bartail flathead (Platycephalus indicus (Linnaeus, 1785))in the Iranian waters of Persian Gulf using the morphometric and meristic characters and by AFLP marker. . A total 180 fish specimens were collected by gill net from six station(khor mosa, bahrekan, shif, motaf, charak and bandar abbas) that was 30 individual related to every station in Iranian shores of Persian Gulf . 28 morphometric factors and 11meristic specialties were measured and morphometric factors was standardized with Beacham formula. Univariate analysis of variance (One-way ANOVA) revealed significant differences with varying degrees between the means for 21 standardized morphometric measurements and 6 meristic counts that showed high significant differences between the six stations sampling. Discriminate function analysis (DFA) or the overall random assignment of individuals into their original groups was for morphometric and meristic characters was 47.9% and 53.9% respectively. The data were subjected to a principle component analysis (PCA) which grouped in eight and four factors for morphometric and meristic charactersrespectively.. Genetic diversity of six populations of bartail flathead (Platycephalus indicus) was investigated using amplified fragment length polymorphism (AFLP). A total of 118 reproducible bands amplified with ten AFLP primer combinations were obtained from 42 fishes that were collected from six different locations in the northern of Persian Gulf. The percentage of polymorphic bands was 57.06%. Average of Nei’s genetic diversity was 0.200±0.008, and Average of Shannon’s index was 0.300±0.011. The results of AMOVA analysis indicated that 66% of the genetic variation contained within populations and 34% occurred among populations and gene flow was 0.6454.The estimated level of population differentiation asmeasured by average Fst value across all loci was 0.327. Plotting discriminant functions 1 and 2 and UPGMA dendrograms based on Euclidian distance and genetic distance also showed at least five separate populations of bartail flathead in the northern Persian Gulf.

Effects of Zataria multiflora Boiss essential oil and nisin on the growth of Staphylococcus aureus in light salted silver carp fillet (Hypophthalmichthys molitrix)

There is an increasing demand in developing newer and safer methods in preserving food products.Among which herbal additives seem to attract evermore attention recently.the major advantage of herbal additives is due to their favorable aroma besides their antimicrobial effects and less expensive than chemical additives. Zataria multiflora Boiss is a native Iranian herb which is used vastly as a food preserver essential oils and also medical usage. Metabolites of harmless bacteria, such as Nisin are also known to be safe preservatives that have antimicrobial activity. However to establish the usefulness of natural antimicrobial preservatives, they must be evaluated alone and in combination with other preservation factors to determine whether there are synergistic effects in rigid media . In this study were evaluated the effects of different concentrations of Zataria multiflora (EO 0, 0.005, 0.015, 0.045, 0.135, 0.405 ,0.810 %) and Nisin(0, 0.15, 0.25, 0.75 μg/ml) and Storage time (up to 21 days) on growth of Staphylococcus aureus ATCC 6538 in a food model system(light salted fish of silver carp, Hypophthalmichthys molitrix). The results on growth of S. aureus were evaluated using SPSS 15.0 statistical software (SPSS 15.0 for windows, SPSS Inc.) and analyzed the logarithm of total count of the bacteria by Tukey Test. Results were considered statistically significant when P≤0.05. The growth of Staphylococcus aureus was affected significantly(P<0.05) by EO and Nisin and also combinations of EO and Nisin. Samples treated with 0.135, 0.405 and 0.810% of thyme essential oil showed a significant decrease on the growth of the bacteria compared with an treated samples(P<0.05). No significant difference was seen on the growth of S.aureus in samples treated with lower concentrations of Z.multiflora(below 0.045%) and untreated group(P>0.05). The most inhibitory effects were seen in samples treated with 0.405% and 0.810% of thyme essential oil until 9 and 12 days after storage,respectively. Also there was significant inhibtory effect(P<0.05) in different concentration of nisin on the organism compared with an treated samples. The synergism effects of the Eo and nisin on the growth rate of the bacteria was significant (P<0.05) compared with untreated samples and samples treated with the Eo or nisin, only. Synergismic effects was observed at concentration of 0.405 and 0.810% of Z. multiflora essential oil with 0.25 μg/ml Nisin, respectively until 15 days after storage. As expected it is preferred to apply the least possible amounts of additives in food preserving that not only are effective and safe but are economically justifiable.