Production of beta-fructofuranosidases by Aspergillus niveus using agroindustrial residues as carbon sources: Characterization of an intracellular enzyme accumulated in the presence of glucose

The production of beta-fructofuranosidases by Aspergillus niveus, cultivated under submerged fermentation using agroindustrial residues, was investigated. The highest productivity of beta-fructofuranosidases was obtained in Khanna medium supplemented with sugar cane bagasse as carbon source. Glucose enhanced the production of the intracellular enzyme, whereas that of the extracellular one was decreased. The intracellular beta-fructofuranosidase was a trimeric protein of approximately 141 kDa (gel filtration) with 53.5% carbohydrate content, composed of 57 kDa monomers (SDS-PAGE). The optimum temperature and optimum pH were 60 degrees C and 4.5, respectively. The purified enzyme showed good thermal stability and exhibited a half-life of 53 min at 60 degrees C. beta-Fructofuranosidase activity was slightly activated by Cu(2+), Mn(2+), Mg(2+), and Na(+) at 1 mM concentration. The enzyme hydrolyzed sucrose, raffinose, and inulin, with K(d) values of 5.78 mM, 5.74 mM, and 1.74 mM, respectively. (C) 2008 Elsevier Ltd. All rights reserved.

Biochemical properties of an extracellular beta-D-fructofuranosidase II produced by Aspergillus phoenicis under Solid-Sate Fermentation using soy bran as substrate

The filamentous fungus A. phoenicis produced high levels of beta-D-fructofuranosidase (FFase) when grown for 72 hrs under Solid-State Fermentation (SSF), using soy bran moistened with tap water (1:0.5 w/v) as substrate/carbon source. Two isoforms (I and II) were obtained, and FFase II was purified 18-fold to apparent homogeneity with 14% recovery. The native molecular mass of the glycoprotein (12% of carbohydrate content) was 158.5 kDa with two subunits of 85 kDa estimated by SDS-PAGE. Optima of temperature and pH were 55 degrees C and 4.5. The enzyme was stable for more than 1 hr at 50 degrees C and was also stable in a pH range from 7.0 to 8.0. FFase II retained 80% of activity after storage at 4 degrees C by 200 hrs. Dichroism analysis showed the presence of random and beta-sheet structure. A. phoenicis FFase II was activated by Mn(2+), Mg(2+) and Co(2+), and inhibited by Cu(2+), Hg(2+) and EDTA. The enzyme hydrolyzed sucrose, inulin and raffinose. K(d) and V(max) values were 18 mM and 189 U/mg protein using sucrose as substrate.

Mitochondrial genome data alone are not enough to unambiguously resolve the relationships of Entognatha, Insecta and Crustacea sensu lato (Arthropoda)

An analysis of the relationships of the major arthropod groups Was undertaken using mitochondrial genome data to examine the hypotheses that Hexapoda is polyphyletic and that Collembola is more closely related to branchiopod crustaceans than insects. We sought to examine the sensitivity of this relationship to outgroup choice, data treatment. gene choice and optimality criteria used in the phylogenetic analysis of mitochondrial genome data. Additionally we sequenced the mitochondrial genome of ail archaeognathan, Nesomachilis australica. to improve taxon selection in the apterygote insects, a group poorly represented in previous mitochondrial phylogenies. The sister group of the Collembola was rarely resolved in our analyses with a significant level of support. The use of different outgroups (myriapods, nematodes, or annelids + mollusks) resulted in many different placements of Collembola. The way in which the dataset was coded for analysis (DNA, DNA with the exclusion of third codon position and as amino acids) also had marked affects on tree topology. We found that nodal Support was spread evenly throughout the 13 mitochondrial genes and the exclusion of genes resulted in significantly less resolution in the inferred trees. Optimality criteria had a much lesser effect on topology than the preceding factors; parsimony and Bayesian trees for a given data set and treatment were quite similar. We therefore conclude that the relationships of the extant arthropod groups as inferred by mitochondrial genomes are highly vulnerable to outgroup choice, data treatment and gene choice, and no consistent alternative hypothesis of Collembola's relationships is supported. Pending the resolution of these identified problems with the application of mitogenomic data to basal arthropod relationships, it is difficult to justify the rejection of hexapod monophyly, which is well supported on morphological grounds. (c) The Willi Hennig Society 2004.

An evaluation of the evidence for linkages between mangroves and fisheries: A synthesis of the literature and identification of research directions

There is a widely held paradigm that mangroves are critical for sustaining production in coastal fisheries through their role as important nursery areas for fisheries species. This paradigm frequently forms the basis for important management decisions on habitat conservation and restoration of mangroves and other coastal wetlands. This paper reviews the current status of the paradigm and synthesises the information on the processes underlying these potential links. In the past, the paradigm has been supported by studies identifying correlations between the areal and linear extent of mangroves and fisheries catch. This paper goes beyond the correlative approach to develop a new framework on which future evaluations can be based. First, the review identifies what type of marine animals are using mangroves and at what life stages. These species can be categorised as estuarine residents, marine-estuarine species and marine stragglers. The marine-estuarine category includes many commercial species that use mangrove habitats as nurseries. The second stage is to determine why these species are using mangroves as nurseries. The three main proposals are that mangroves provide a refuge from predators, high levels of nutrients and shelter from physical disturbances. The recognition of the important attributes of mangrove nurseries then allows an evaluation of how changes in mangroves will affect the associated fauna. Surprisingly few studies have addressed this question. Consequently, it is difficult to predict how changes in any of these mangrove attributes would affect the faunal communities within them and, ultimately, influence the fisheries associated with them. From the information available, it seems likely that reductions in mangrove habitat complexity would reduce the biodiversity and abundance of the associated fauna, and these changes have the potential to cause cascading effects at higher trophic levels with possible consequences for fisheries. Finally, there is a discussion of the data that are currently available on mangrove distribution and fisheries catch, the limitations of these data and how best to use the data to understand mangrove-fisheries links and, ultimately, to optimise habitat and fisheries management. Examples are drawn from two relatively data-rich regions, Moreton Bay (Australia) and Western Peninsular Malaysia, to illustrate the data needs and research requirements for investigating the mangrove-fisheries paradigm. Having reliable and accurate data at appropriate spatial and temporal scales is crucial for mangrove-fisheries investigations. Recommendations are made for improvements to data collection methods that would meet these important criteria. This review provides a framework on which to base future investigations of mangrove-fisheries links, based on an understanding of the underlying processes and the need for rigorous data collection. Without this information, the understanding of the relationship between mangroves and fisheries will remain limited. Future investigations of mangrove-fisheries links must take this into account in order to have a good ecological basis and to provide better information and understanding to both fisheries and conservation managers.

Evolutionary transition to freshwater by ancestral marine palaemonids: evidence from osmoregulation in a tide pool shrimp

The transition from marine/brackish waters to freshwater habitats constitutes a severe osmotic and ionic challenge, and successful invasion has demanded the selection of morphological, physiological, biochemical and behavioral adaptations. We evaluated short-term (1 to 12 h exposure) and long-term (5 d acclimation), anisosmotic extracellular (osmolality, [Na(+), Cl(-)]) and long-term isosmotic intracellular osmoregulatory capability in Palaemon northropi, a neotropical intertidal shrimp. F northropi survives well and osmo- and ionoregulates strongly during short- and long-term exposure to 5-45 parts per thousand salinity, consistent with its rocky tide pool habitat subject to cyclic salinity fluctuations, Muscle total free amino acid (FAA) concentrations decreased by 63% in shrimp acclimated to 5%. salinity, revealing a role in hypoosmotic cell volume regulation; this decrease is mainly a consequence of diminished glycine, arginine and proline. Total FAA contributed 31% to muscle intracellular osmolality at 20 parts per thousand, an isosmotic salinity, and decreased to 13% after acclimation to 5 parts per thousand. Gill and nerve tissue FAA concentrations remained unaltered. These tissue-specific responses reflect efficient anisosmotic and anisoionic extracellular regulatory mechanisms, and reveal the dependence of muscle tissue on intracellular osmotic effectors. FAA concentration is higher in P. northropi than in diadromous and hololimnetic palaemonids, confirming muscle FAA concentration as a good parameter to evaluate the degree of adaptation to dilute media. The osmoregulatory capability of P. northropi may reflect the potential physiological capacity of ancestral marine palaemonids to penetrate into dilute media, and reveals the importance of evaluating osmoregulatory processes in endeavors to comprehend the invasion of dilute media by ancestral marine crustaceans.

Measurement of IgE antibodies to shrimp tropomyosin is superior to skin prick testing with commercial extract and measurement of IgE to shrimp for predicting clinically relevant allergic reactions after shrimp ingestion

Background: Shrimp is a frequent cause of food allergy. Tropomyosin is the major allergen in shrimp, and it shares homology to tropomyosins from other crustaceans, dust mites, cockroach, and parasites. Objective: The aim of this study was to determine the value of detection of IgE to shrimp tropomyosin in the diagnosis of shrimp allergy. Methods: We have studied 35 patients with asthma, rhinitis, or both who were sensitized to Dermatophagoides pteronyssinus. All subjects underwent skin prick testing in addition to double-blind, placebo-controlled food challenges (DBPCFC); oral open challenges; or both with shrimp. Measurements of IgE to shrimp and shrimp tropomyosin were carried out by means of CAP and chimeric ELISA, respectively. Results: Oral challenges confirmed the diagnosis of shrimp allergy in 7 patients. IgE measurement to shrimp tropomyosin was positive in 71.4% of the patients with shrimp allergy. Of the 28 patients without shrimp allergy, only 7.1% (2/28) had IgE to shrimp tropomyosin compared with 25% (7/28) who had IgE to shrimp and 35.7% (10/28) who had positive skin prick test responses to shrimp. Sensitivity was similar for all 3 methods (71.4%); in contrast, specificity of IgE to shrimp tropomyosin (92.8%) was greater than that of IgE to shrimp (75%) and skin prick testing (64.2%). With regard to diagnostic efficiency, measurement of IgE to shrimp tropomyosin was superior to measurement of IgE to shrimp and skin prick testing (88.5%, 74.2%, and 65.7%, respectively). Conclusion: Use of measurements of IgE to shrimp tropomyosin provided added value to the diagnosis of shrimp allergy. (J Allergy Clin Immunol 2010;125:872-8.)

Echinobothrium chisholmae n. sp (Cestoda, Diphyllidea) from the giant shovel-nose ray Rhinobatos typus from Australia, with observations on the ultrastructure of its scolex musculature and peduncular spines

Echinobothrium chisholmae n. sp. is described from Rhinobatos typus Bennett (Rhinobatidae), collected from Heron Island, Great Barrier Reef, Australia. E. chisholmae differs from all congeners in possessing 11 hooks in each dorsal and ventral group on the rostellum and groups of 3-6 hooklets on either side of the hooks. A single metacestode of E. chisholmae was collected from the decapod crustacean Penaeus longistylus Kubo. Yellow pigmentation of the cephalic peduncle in immature adults is caused by the accumulation of large vesicles in the distal cytoplasm of the tegument. The vesicles probably provide materials for spine formation. Ultrastructural examination of the rostellar musculature revealed that the muscles are stratified (striated-like), consisting of a periodic repetition of sarcomeres separated by perforated Z-like lines that are oblique to the long axes of the myofilaments.

Morphological, Morphometrical and Histochemical Study of the Lining and Glandular Epithelia of the Tongue of the Bullfrog Rana catesbeiana

The dorsal surface of the tongue of the bullfrog, Rana catesbeiana, has simple columnar epithelium with a few ciliated cells and goblet cells. The entire surface is covered with numerous filiform papillae and few fungiform. Filiform papillae have a simple columnar epithelium with secretory cells, while the fungiform have a sensory disc on their upper surface the lined by a stratified columnar epithelium with basal, peripheral, glandular and receptor cells. Over the dorsal lingual surface there are numerous winding tubular glands, which penetrate deeply into the muscle of the tongue, mingling with the fibers. The gland epithelium is cylindrical with secretory and supporting cells. The first are absolute on the basis of the gland and the latter are rare in the upper third. The ventral surface of the tongue is lined by a stratified epithelium, with the presence of goblet cells, with ciliated cells among them. Morphometrically, lingual glands varies in length, according to their location: shorter in the anterior region of the tongue (330 mu m) than in the posterior region (450 mu m). Secretory cells of the anterior lingual glands are smaller (1457.7 mm(3)) than the posterior ones (2645.9 mu m(3)). The same can be said of the cell nuclei, 130.0 mu m(3) for the anterior glands and 202.3 mu m(3) for the posterior ones. Secretory cells of the lingual glands contain substances rich in protein and neutral mucopolysaccharides, which characterize the seromucous type. Goblet cells of the dorsal and ventral surface epithelia secrete neutral mucopolysaccharides and proteins, and can be characterized as type G1 cells, and the supporting cells of the superficial glands of the fungiform papillae secrete a mucus rich in neutral mucopolysaccharides, sulfomucins and sialomucins.

Polycyclic aromatic hydrocarbon levels in three pelagic fish species from Atlantic Ocean: inter-specific and inter-season comparisons and assessment of potential public health risks

The concentrations of 18 polycyclic aromatic hydrocarbons (PAHs) were determined in three commercially valuable fish species (sardine, Sardina pilchardus; chub mackerel, Scomber japonicus; and horse mackerel, Trachurus trachurus) from the Atlantic Ocean. Specimens were collected seasonally during 2007–2009. Only low molecular weight PAHs were detected, namely, naphthalene, acenaphthene, fluorene and phenanthrene. Chub mackerel (1.80–19.90 microg/kg ww) revealed to be significantly more contaminated than horse mackerel (2.73–10.0 microg/kg ww) and sardine (2.29–14.18 microg/kg ww). Inter-specific and inter-season comparisons of PAHs bioaccumulation were statistically assessed. The more relevant statistical correlations were observed between PAH amounts and total fat content (significant positive relationships, p < 0.05), and season (sardine displayed higher amounts in autumn–winter while the mackerel species showed globally the inverse behavior). The health risks by consumption of these species were assessed and shown to present no threat to public health concerning PAH intakes.

Lipid content of frozen fish: comparison of different extraction methods and variability during freezing storage

In this work, a microwave-assisted extraction (MAE) methodology was compared with several conventional extraction methods (Soxhlet, Bligh & Dyer, modified Bligh & Dyer, Folch, modified Folch, Hara & Radin, Roese-Gottlieb) for quantification of total lipid content of three fish species: horse mackerel (Trachurus trachurus), chub mackerel (Scomber japonicus), and sardine (Sardina pilchardus). The influence of species, extraction method and frozen storage time (varying from fresh to 9 months of freezing) on total lipid content was analysed in detail. The efficiencies of methods MAE, Bligh & Dyer, Folch, modified Folch and Hara & Radin were the highest and although they were not statistically different, differences existed in terms of variability, with MAE showing the highest repeatability (CV = 0.034). Roese-Gottlieb, Soxhlet, and modified Bligh & Dyer methods were very poor in terms of efficiency as well as repeatability (CV between 0.13 and 0.18).

Analysis of polycyclic aromatic hydrocarbons in fish: evaluation of a quick, easy, cheap, effective, rugged, and safe extraction method

QuEChERS method was evaluated for extraction of 16 PAHs from fish samples. For a selective measurement of the compounds, extracts were analysed by LC with fluorescence detection. The overall analytical procedure was validated by systematic recovery experiments at three levels and by using the standard reference material SRM 2977 (mussel tissue). The targeted contaminants, except naphthalene and acenaphthene, were successfully extracted from SRM 2977 with recoveries ranging from 63.5–110.0% with variation coefficients not exceeding 8%. The optimum QuEChERS conditions were the following: 5 g of homogenised fish sample, 10 mL of ACN, agitation performed by vortex during 3 min. Quantification limits ranging from 0.12– 1.90 ng/g wet weight (0.30–4.70 µg/L) were obtained. The optimized methodology was applied to assess the safety concerning PAHs contents of horse mackerel (Trachurus trachurus), chub mackerel (Scomber japonicus), sardine (Sardina pilchardus) and farmed seabass (Dicentrarchus labrax). Although benzo(a)pyrene, the marker used for evaluating the carcinogenic risk of PAHs in food, was not detected in the analysed samples (89 individuals corresponding to 27 homogenized samples), the overall mean concentration ranged from 2.52 l 1.20 ng/g in horse mackerel to 14.6 ± 2.8 ng/ g in farmed seabass. Significant differences were found between the mean PAHs concentrations of the four groups.

Reproductive aspects of Macrobrachium amazonicum (Decapoda: Palaemonidae) in the State of Amapá, Amazon River mouth

Macrobrachium amazonicum is an indigenous prawn vastly distributed in basins of South America, widely exploited by artisanal fisheries in northern and northeastern Brazil and, with great potential for aquaculture. This study aimed to investigate general aspects of population structure and reproductive characteristics (size at first maturity, fecundity and reproductive output) of M. amazonicum from two important areas to artisanal prawn fishing located at the mouth of the Amazon River, State of Amapá. The specimens were captured using 20 handcrafted traps called "matapi". A number of 5,179 prawns were captured, 2,975 females and 2,195 males resulting in 1.35:1 female to male ratio. Santana Island and Mazagão Velho showed females predominated in the population. A reproductive peak period was observed from January to April/2009 and in December/2010, coinciding with the period of higher rainfall. The recruitment peak occurred in June and July/2009. Egg-bearing females ranged in size (carapace length) from 11.10 to 29.6 mm. Fecundity increased with female size and reached up to 7,417 eggs. This amount of eggs is considered low if compared with other Macrobrachium estuarine species. Mean egg volume increased gradually from 0.121 to 0.24 mm³ during embryogenesis, representing 68.5% of overall increase from Stage I to Stage III. Eggs of M. amazonicum are small; this is typical for Macrobrachium species, which depends on brackish water to complete the larval development. Irrespective of female size, reproductive output of M. amazonicum varied between 4.8 and 21.85% of their body weight into eggs production.

Reproductive biology of Macrobrachium surinamicum (Decapoda: Palaemonidae) in the Amazon River mouth

Macrobrachium surinamicum is an indigenous prawn distributed from the lower Amazon and Tocantins river basins to Venezuela in the Orinoco Delta region. It is common bycatch fauna of Macrobrachium amazonicum artisan fishing in the states of Pará and Amapá. The aim of this study was to investigate aspects on reproductive biology (reproductive period, size of sexual maturity population, fecundity, reproductive output and recruitment) of M. surinamicum from four important areas to artisanal prawn fishing located at the Amazon River mouth (Amapá and Pará). The specimens were captured using 20 handcrafted traps called "matapi". A number of 675 prawns were captured, 258 males, 409 females and eight juveniles, resulting in 1:1.6 (Male: Female) sex ratio. The reproductive peak period occurred from March to July, coinciding with the higher rainfall period. The juvenile prawn occurred only in May and July. Total length of egg-bearing females ranged from 12.12 to 38.30 mm, with mean female length at first maturity (L50) of 23.7 mm. Fecundity increased with prawn size and varied between 174 and 1780 eggs per female. Mean egg volume increased gradually from 0.031 (Stage I) to 0.060 mm³ (Stage III) during embryogenesis. Macrobrachium surinamicum depends on brackish water to complete the larval development. Irrespective of female size, reproductive output of M. surinamicum varied between 4.3 % and 35.5 % of their body weight for egg production. The knowledge of the reproductive biology reported in the present study is an important tool to define strategies to preserve M. surinamicum in Amazon River mouth.

Foregut morphology of Macrobrachium carcinus (Crustacea, Decapoda, Palaemonidae)

ABSTRACT Macrobrachium carcinus is a Brazilian native prawn with recognized potential for use in aquaculture activities. The aim of this study was to describe and illustrate in detail the morphology of the M. carcinus foregut. The foregut comprises the mouth, esophagus and stomach. It is lined by a simple cylindrical epithelium overlain by chitinous cuticle. The cardiac chamber is well supplied with muscles and lined with chitin thickened in places to form a complex, articulating set of ossicles. The ossicles and setae inside the cardiac chamber seem to direct the food movement through the cardiac chamber and sort the food according to particle size as digestion takes place. Twenty-one basic ossicles were observed in the stomach ofM. carcinus and are divided into seven categories, reflecting their presumed functional roles. The significance of these morphological features is discussed in terms of its implication in feeding management that can support future commercial farms of this important fishery resource.

Population genetic structure of brown crab (Cancer pagurus) in Irish waters

The brown crab (Cancer pagurus) fishery in Ireland is one of the most important financially and socio-economically, with the species worth approximately €15m per year in the first half of the decade. Only mackerel (Scomber scombrus) and Dublin Bay prawn (Nephrops norvegicus) are of greater value. Despite this, very little research has been conducted to describe the stock structure of brown crab on a national scale. In this study a country-wide assessment of genetic population structure was carried out. Sampling was conducted from commercial fishing boats from 11/06 to 04/08 at seven sample sites representing the central Irish brown crab fisheries, with one sample site from the UK also included in the study. Six microsatellite markers, specifically developed for brown crab, were used to assess genetic diversity and estimate population differentiation parameters. Significant genetic structuring was found using F-statistics (Fst = 0.007) and exact tests, but not with Bayesian methods. Samples from the UK and Wexford were found to be genetically distinct from all other populations. Three northern populations from Malm Head and Stanton Bank were genetically similar with Fst estimates suggesting connectivity between them. Also, Stanton Bank, again on the basis of Fst estimates, appeared to be connected to populations down the west coast of Ireland, as far south as Kerry. Two Galway samples, one inside and one outside of Galway Bay, were genetically differentiated despite their close geographic proximity. It is hypothesised that a persistent northerly summer current could transport pelagic larvae from populations along the southwest and west coasts of Ireland towards Stanton Bank in the North, resulting in the apparent connectivity observed in this study.