Stimulating effects of pond water on digestive enzyme activity in the Pacific white shrimp, Litopenaeus vannamei (Boone)

Crude enzyme extracts were obtained from the digestive glands of Pacific white shrimp, Litopenaeus vannamei (Boone), reared in oligotrophic well water and eutrophic shrimp pond water to compare digestive enzyme activity between the two groups. Specific activities of serine protease, collagenase, amylase, cellulase, lipase and acid phosphatase were significantly higher (P < 0.01) in pond water‐reared shrimp (PW shrimp) than in well water reared‐shrimp (WW shrimp). For most enzymes assayed, specific activity was more than two times higher in PW shrimp, and cellulase activity was over six times higher. In contrast, chitinase activity was significantly higher (P < 0.001) in WW shrimp. Higher specific activity of most digestive enzymes in PW shrimp was probably due to natural productivity in the pond water that served as a source of organic substrates, and this increased activity may contribute to the growth‐enhancing effect of shrimp pond water.     

Note on the methods for determination of chromic oxide in shrimp feeds.

Chromic oxide is used as an inert marker to measure apparent digestibility of feeds in insects, terrestrial, and aquatic animals. Quantitative determination of chromic oxide content in the sample requires the oxidation of water insoluble trivalent chromic oxide to its water-soluble hexavalent form. The two commonly used oxidizing agents are 70% perchloric acid or a mixture of sodium molybdate, sulfuric, and perchloric acid. Chromic oxide content of the oxidized solution is then measured against known standards either directly by spectrophotometry in the visible range at 350, 370, or 440 nm or after forming a colored complex with diphenylcarbazide (DPC) by colorimetry at 540 nm. This study compared the two methods of oxidation followed by spectrophotometry at the three wavelengths and by DPC colorimetry. DPC colorimetry gave precise results than the direct measurement of dichromate ion, irrespective of the method of oxidation used. Ash from samples oxidized by perchloric acid and quantified by DPC colorimetry gave a better measure of actual chromic oxide content as compared to the other methods tested.     

Genetic variations and interrelationships in Vanilla planifolia and few related species as expressed by RAPD polymorphism

Vanilla is naturally distributed in Mexico and parts of Central America and the history of origin of cultivated vanilla suggests that the entire stock outside Mexico may be from a single genetic source. In the present study, RAPD polymorphism was used to estimate the level of genetic diversity and interrelationships among different collections of Vanilla planifolia Andr., and few related species, including both leafy and leafless types such as V. tahitensis J.W.Moore, V. andamanica, Rolfe, V. pilifera Holtt., and V. aphylla Blume. Studies revealed that there are very limited variation within collections of V. planifolia, indicative of its narrow genetic base, and of the related species we tested, V. tahitensis is nearest to V. planifolia. The species studied are diverse and have a similarity ranging from 1.2 to 57.3 %. Of the sampled taxa, V. andamanica is the most divergent and there is also reasonable variability within its collections, indicating the possibility of natural seed set. A total of 82 polymorphic bands expressed in the RAPD profiles were used to generate a genetic distance matrix, which was then used in cluster analysis. Specific groupings were revealed by the cluster analysis whereby the leafless forms (V. aphylla, V. pilifera and the new species) and V. andamanica formed separate clusters. This is the first report of species interrelationship studies, including both cultivated and wild vanilla species.     

Antimicrobial and cellular activity of poly(L-lactide)/chitosan/Imipenem antibiotic composite nanofibers

Synthesis of biocompatible polymer nanofibers is valuable, due to their use as a cover for burns and as a replacement for bandage because of their antimicrobial properties. In this study, electrospinning of chitosan(Ch) and nanofibers synthesis with antibacterial properties was investigated. Nanofibers with antibacterial properties were synthesized by electrospun of Ch/poly(L-lactide)(PLA)/Imipenem(Imi) polymer solution. The results showed that the optimized ratio of Ch/PLA polymer solution was ratio of 50:50 and Ch 2 wt% and PLA 10 wt% polymer solution was the best weight percentage for nanofiber preparation. Also, the average diameter of Ch/PLA/Imi nanofibers was 143 nm and measured with ImageJ software. Afterwards, the antibacterial properties of Imi as additives (with different percentages) was studied in the polymer solution. The scanning electron microscopy (SEM) images and antibacterial tests were showed that the electrospun of Ch/PLA/Imi polymeric nanofibers were effective against Gram negative bacteria Escherichia coli (E. coli) and inhibited growth of E. coli. The growth and viability percentage of fibroblast cells with nanofibers in αMEM culture are at desirable levels after 6 days.     

Role of nitric oxide on the generation of atretic follicles in the rat ovaries

The aim of this study was to investigate the role of Nitric Oxide (NO) in the atresia of ovarian follicles in an animal model. Twenty adult, female rats (90 days old with body weights of210 +/- 10 g in the beginning of the experiments) were divided into 4 groups of 5 each. They were treated twice daily from the subcutaneous route for 21 successive days with either of the following chemicals: nitroglycerine, L-arginine, L-NAME, or saline. On day 22, all animals were sacrificed. Ovaries were dissected out free of connected tissue and were fixed in formaline 10%. Later, paraffine blocks were prepared and serial sections were made by means of H and E routine staining method. Intact and atretic follicles were counted separately. In addition, damages were analyzed qualitatively from the points of view of appearance and morphologic changes. In the evaluation of ovarian follicular structures, different types of healthy as well as atretic follicles were observed. In most of atretic follicles, the oocytes were abnormally elongated and increnation of their outlines were obvious. There were numerous macrophages around and inside of the atretic follicles. Our investigation regarding the distribution of atretic follicles in the ovaries of test groups revealed that atretic follicles in the L-NAME treated group were increased in comparison to the control group. Conversely, however, in the arginine-treated group, the atretic follicles were reduced compared to the control animals. Treatment with nitroglycerine of the rats decreased the number of atretic follicles significantly (p < 0.05) in comparison to the control group. In conclusion, enhanced NO, either from endogenous or exogenous origins, prevents atresia phenomenon, while inhibition of NO exerts an opposite effect.     

Method for determining the physical stability of shrimp feeds in water

Three methods for measuring the water stability and leaching characteristics of shrimp pelleted feeds were developed and tested: static water method, horizontal shaking method and vertical shaking method. Each method was tested using a commercial and experimental pelleted shrimp feed. Both feeds had a pellet size of 2.4 mm × 5.0 mm with similar proximate composition (35% protein and 9% fat). Each test run consisted of leaching c. 2 g of feed at six intervals of immersion time (0, 30, 60, 120, 240, 360 min), two concentrations of salinity (0, 34 ppt), and three different water temperatures (15, 25, 35 °C). A refrigerated circulating water bath was used to maintain the desired level of water temperature. For the static water and horizontal shaking methods, a Buchner filtration apparatus with Whatman filter paper no. 3 (5 μ) was used to separate the remaining pellets from leached water. The vertical shaking method (VanKel) utilized a perforated stainless‐steel basket (381 μ) for leaching and serving as a filter to separate pellets from leached water. For all three methods, the leached feed and original feed samples were dried in a convection oven at 105 °C for 24 h and then cooled in a desiccator. Dried feed samples were weighed and analysed for dry matter retention. Pellet stability was calculated as the ratio of dry matter retention after leaching and dry matter of original samples expressed as a percentage. Results indicated that any method for pellet stability analysis should take into account water temperature and salinity in addition to actual pellet agitation rate to obtain accurate and consistent measurement of pellet disintegration and nutrient leaching. Dry matter retention was different for each shrimp feed with experimental feed (72.8%) showing lower retention than commercial feed (88%) after 6 h of VanKel leaching under normal shrimp culture conditions. The three methods provided reproducible results with both horizontal and vertical shaking methods recommended for routine laboratory analysis because they could provide different degrees of pellet agitation that simulate actual indoor and outdoor shrimp culture conditions.     

Digestive enzymes present in Pacific threadfin Polydactylus sexfilis (Bloch & Schneider 1801) and bluefin trevally Caranx melampygus (Cuvier 1833)

Pacific threadfin Polydactylus sexfilis (Bloch & Schneider 1801) and bluefin trevally Caranx melampygus (Cuvier 1833) are warmwater marine finfish currently under development for aquaculture in the Pacific. Differences in specific activities of digestive enzymes extracted from the stomach and mid-gut were compared to gain insight into their feeding habits in the wild and to understand their nutritional needs. Adult fish were maintained in captivity and fed a commercial pelleted feed. Serine protease measured in all tissues was at least 20 times higher in threadfin than in trevally. Aspartic proteases were the major digestive enzymes found in trevally. There was a 34-fold increase in collagenase activity in the intestine of threadfin from the prefed to the fed state. Chitinase activity was found in the stomach, pylorus and intestine of both species. However, specific activity in pylorus and intestine of threadfin increased 2.75 and 4 times, respectively, but showed little change in trevally. Amlyases were found only in trevally. Increase in lipase specific activity in the gut of trevally was higher than that for threadfin. The results indicated that the two species have diverse digestive capabilities. This appears to be consistent with their feeding habits in nature. Threadfin are more adapted to a wider range of food protein sources than trevally, but appear to be less well adapted than trevally to using complex carbohydrates. These observations may provide a basis for practical diet formulations for these two species.     

Interspecific hybridization in vanilla and molecular characterization of hybrids and selfed progenies using RAPD and AFLP markers

Vanilla, Vanilla planifolia Andrews, is native to Mexico and Central America, but is now cultivated in other parts of the tropics. Continuous clonal propagation has resulted in very little variability for crop improvement programmes in vanilla. In this study, an attempt has been made to increase the spectrum of variation by interspecific hybridization with Vanilla aphylla, an Indian species which is tolerant to Fusarium. Interspecific hybrids were successfully produced and morphological characters and molecular profiles revealed the true hybridity of the progenies. Ten seedling progenies of V. planifolia, and four interspecific hybrids obtained from crosses between V. planifolia (female) and V. aphylla (male) using a number of different loci as markers were evaluated and 319 amplified fragment length polymorphisms (AFLPs) and 83 random amplified polymorphic DNAs (RAPDs) loci were marked. The profiles indicate similarity between the parents, selfed progenies and interspecific hybrids and that all the progenies tested were variable when compared to each other, which can be exploited for crop improvement in vanilla. This is the first report in vanilla, indicating that RAPD and AFLP profiles coupled with morphological characters can be utilized to assess the variability and hybrid nature of genotypes and of successful interspecific hybridization and production of hybrids between V. planifolia and V. aphylla.     

The amino acid and fatty acid compositions of selected tissues of the dolphin fish (Coryphaena hippurus) and their nutritional implications

The amino acid and fatty acid compositions of muscle, gill, intestine, liver, ovaries, and whole bodies from captive juvenile and adult, and wild adult dolphin fish (Coryphaena hippurus) were determined. Essential amino acid ratios (A/E ratios) of the whole body and individual tissues were calculated and compared. In addition, the concept of a metabolic A/E ratio calculated from known daily protein turnover rates and from the amino acid profiles is introduced. Fatty acid profiles of phospholipid (PL) and triglyceride (TG) fractions in muscle and liver were also compared.

There were few differences in the amino acid and fatty acid profiles of tissues between the different groups of dolphin fish examined. Linolenic series fatty acids were quantitatively the most important polyunsaturates in tissue PL fractions. C22: 6n−3 was the major polyunsaturate present, accounting for approximately 20% of the total PLs in muscle and liver. There was no significant difference in tissue essential amino acid patterns between adult fish raised in captivity, and adult fish caught in the wild. However, the muscle of juvenile fish was lower in arginine and histidine contents than the muscle of adult fish. Ovaries had the lowest arginine, histidine, and tryptophan contents of all tissues measured.

Calculated A/E ratios of the muscle and whole body, and A/E ratios based on the metabolic rates for protein turnover in muscle, gill, and intestine, were similar. All ratios predicted an ontological increase in dietary arginine, histidine, and tryptophan needs of dolphin fish consistent with the changes noted in tissue amino acid contents.

It is concluded that the calculated A/E ratios and fatty acid profiles of dolphin fish tissues may be useful guides in formulating diets for the culture of this species. The results obtained suggest that the requirements for arginine, histidine, and tryptophan may differ between subadult and adult fish. Dietary needs for linolenic series HUFAs, especially C22:6n−3, are likely high throughout the life of this fish.