EVALUATION OF POTENTIAL SUBSTRATES TO MONITOR RESPIRATORY NITRATE REDUCTASE ACTIVITY IN SOILS

Different inorganic ions were tested for their ability to replace nitrate as the terminal oxidant for bacterial respiration in anaerobic soils. Chlorate, bromate. selenate. tellurite and ferric ions were all unsuitable substitutes, lodate reduction in soils was similar to nitrate reduction because it required anaerobic conditions, was increased by glucose amendment and decreased by phenylmercuric acetate. Amendment of soils with iodate and measurement of iodide production can be used to measure nitrate reductase activity in a soil and so give a qualitative indication, and possibly a quantitative estimate, of the denitrification capacity of soils.     

Immunohistochemical study on changes in gill Na+/K+-ATPase α-subunit during smoltification in the wild masu salmon, Oncorhynchus masou

Changes in immunoreactivity of Na⁺/K⁺-ATPase -subunit in gill sections of wild masu salmon (Oncorhynchus masou) during the parr-smolt transformation (smoltification) were compared with changes in gill Na⁺/K⁺-ATPase specific activity. Gill Na⁺/K⁺-ATPase specific activity increased from April and peaked in May. Immunohistochemical analysis, using an antiserum against a synthetic oligopeptide based on the conserved region of the Na⁺/K⁺-ATPase -subunit, revealed that immunoreactivity was confined to chloride cells in the surface layer of primary lamellae and the proximal end of secondary lamellae. The size and number of these cells increased gradually from February to May; however, the number of chloride cells of the secondary lamellae decreased in May. These data suggest that the synthesis of Na⁺/K⁺-ATPase and the proliferation of chloride cells occur prior to the elevation of enzyme activity. Moreover, it is likely the proliferation and hypertrophy of chloride cells on primary lamellae prepare smolts for entry into seawater and migration in the ocean.     

Insulin and IGF-I receptors and tyrosine kinase activity in carp ovaries: changes with reproductive cycle

Insulin and insulin-like growth factor I (IGF-I) receptors from carp ovaries were semipurified with wheat germ agglutinin at different moments of the reproductive cycle and their binding characteristics and tyrosine kinase activity were studied. Specific receptors for insulin and IGF-I were found. IGF-I receptors presented higher binding (23.8 ± 1.5%), number of receptors (965 ± 20fm/mg) and affinity (K_D 0.24 ± 0.03nM) than those shown for insulin receptors (4.1 ± 1%, 530 ± 85fm/mg and 0.85 ± 0.1nM, respectively). Insulin and IGF-I receptors have a tyrosine kinase activity which is not different from that found in muscle of the same species. Seasonal changes were found in binding, with maximum values for insulin and IGF-I reached at the end of pre-spawning period (June). However, while IGF-I binding was observed in all stages, insulin binding decreased in autumn and disappeared in winter, which suggests a different role for the two peptides in ovarian physiology.     

Development and characterization of a cell line TTCF from endangered mahseer Tor tor (Ham.)

Tor tor is an important game and food fish of India with a distribution throughout Asia from the trans-Himalayan region to the Mekong River basin to Malaysia, Pakistan, Bangladesh and Indonesia. A new cell line named TTCF was developed from the caudal fin of T. tor for the first time. The cell line was optimally maintained at 28°C in Leibovitz-15 (L-15) medium supplemented with 20% fetal bovine serum (FBS). The propagation of TTCF cells showed a high plating efficiency of 63.00%. The cytogenetic analysis revealed a diploid count of 100 chromosomes at passage 15, 30, 45 and 60 passages. The viability of the TTCF cell line was found to be 72% after 6 months of cryopreservation in liquid nitrogen (-196°C). The origin of the cell lines was confirmed by the amplification of 578- and 655-bp sequences of 16S rRNA and cytochrome oxidase subunit I (COI) genes of mitochondrial DNA (mtDNA) respectively. TTCF cells were successfully transfected with green fluorescent protein (GFP) reporter plasmids. Further, immunocytochemistry studies confirm its fibroblastic morphology of cells. Genotoxicity assessment of H?O? in TTCF cell line revealed the utility of TTCF cell line as in vitro model for aquatic toxicological studies.     

Fish-based methods for assessing European running waters: a synthesis

Abstract  The European Union, Water Framework Directive (WFD) requires monitoring of riverine fish fauna. When the WFD came into force in 2000, most of the EU member states did not have fish-based assessment methods compliant to WFD requirements. Therefore, the objectives of FAME ( http://fame.boku.ac.at ), a project under the fifth R&D Framework Programme of the European Commission were to develop, evaluate and implement a standardised fish-based method for assessing the ecological status of European running waters. This paper synthesises the outputs of FAME and defines future research needs. Two different methodologies were used: the so-called spatially based modelling and the site-specific modelling, the latter leading to the European Fish Index (EFI). The advantage of the EFI is that, despite being a single index, it is applicable to a wide range of environmental conditions across Europe precluding the need for inter-calibration. The EFI will support the WFD towards harmonised/standardised assessment and management of running waters in Europe, thus enabling comparative analyses of the ecological status of running waters across Europe.     

An investigation into the bacterial aetiology of 'black patch necrosis' in Dover sole, Solea solea L.

Abstract. A recurring disease known as 'black patch necrosis' (BPN) has been the cause of severe mortalities in 0-group and 1-group Dover sole during the period 1974-1978 at the Sea Fish Industry Authority's Marine Cultivation Unit at Hunter-ston, Ayrshire, Scotland. Samples of diseased and healthy Dover sole tissue were examined microbiologically and a long thin filamentous Gram-negative organism, strongly resembling Flexibacter columnar's , was repeatedly isolated from diseased tissue but was not found in healthy tissue. This organism was pathogenic to Dover sole, producing 100% mortality in 96 h at 17.5°C (±2°C) and the organism was reisolated from all the infected fish. The pathogenic isolate and reisolate were examined and compared with reference strain NCMB 1038 of Flexibacter columnarts , formerly known as Chondrococcus columnaris or Cytophaga columnaris. The isolate, reisolate and type culture were identical with respect to cell morphology, oxidase test, catalase production, production of H₂S, casein hydrolysis, gelatin liquefaction, hydrolysis of Tween 20, degradation of tyrosine and starch hydrolysis. All bacterial types showed similar patterns of sensitivity to a range of antibiotics. It is proposed that a Flexibacter columnari's-like organism is associated with the condition in farmed Dover sole known as 'black patch necrosis'.     

Method for Determining Apparent Digestibility of Carbohydrate and Protein Sources for Artificial Diets for Juvenile Sea Cucumber,Australostichopus mollis

Effective nursery diets can greatly reduce the cost of producing juvenile sea cucumbers for release to aquaculture or restocking programs. However, methods for systematically testing the bioavailability of artificial diet ingredients for sea cucumbers are poorly developed, and consequently, there has been little research in this field. The current study presents methods developed to test the suitability of common carbohydrate and protein sources for inclusion in artificial diets for juvenile Australostichopus mollis. Apparent digestibility of carbohydrates was moderate and did not exceed 50% for any carbohydrates assayed. Wheat starch and carrageenans showed the highest digestibility amongst carbohydrates. Differences in apparent digestibility were not reflected in growth performance of juvenile A. mollis fed carbohydrate diets; overall growth performance was poor for all diets. Artificial protein sources consistently exhibited higher apparent digestibility, ranging from 75.1% for fish meal to 98.1% for casein. Low‐cost protein sources, like meat meal, show promise for future use in artificial diet formulation. However, delivering high protein content may reduce ingestion rates and thereby lower overall digestive efficiency in juveniles. Results show that artificial carbohydrate sources have some potential as diet constituents for juvenile sea cucumbers but are unsuitable as primary energy sources. Future testing of artificial carbohydrate sources for A. mollis may require predigestion to improve digestibility.     

n‐3 and n‐6 fatty acid bioconversion abilities in Eurasian perch (Perca fluviatilis) at two developmental stages

The aim of this work was to study the fatty acid (FA) bioconversion ability in Eurasian perch fed with diets differing in their polyunsaturated fatty acids (PUFA) from n‐3 and n‐6 series content at two development stages: adults in exogenous vitellogenesis, and juveniles during the on‐growing phase. Duplicate groups of adults and juveniles were fed for 12 weeks with four diets: D1 and D2, two diets prepared with fish oil partially or totally as the lipid source, and so containing long‐chain PUFA (LC‐PUFA). Those two diets differed by their n‐3/n‐6 FA dietary ratio (0.2 and 7.0, respectively), D1 being characterized by a high n‐6 LC‐PUFA level, while D2 had a high level of n‐3 LC‐PUFA. D3 and D4 were constituted only with vegetable oils, and were therefore devoid of LC‐PUFA. D3 was characterized by a high level of 18:2 n‐6 (n‐6/n‐3 ratio of 0.3), while D4 was characterized by a high level of 18:3 n‐3 (n‐3/n‐6 ratio of 1.9). Both groups of fish were able to elongate and desaturate the 18:3 n‐3 precursor into eicosapentaenoic acid and docosahexaenoic acid, regarding the FA profile of livers. Furthermore, total elongation/desaturation from [1‐¹⁴C]18:3 n‐3 of LC‐PUFA was higher in fish fed with the high dietary 18:3 n‐3 level compared to the diet rich in n‐3 LC‐PUFA. By opposition, the bioconversion of 18:2 n‐6 into LC‐PUFA was limited, regarding the elongation/desaturation activity of LC‐PUFA from [1‐¹⁴C]18:2 n‐6. In view of the great ability for bioconversion of n‐3 FA, linseed oil is a promising alternative to fish oil in formulating feed for juveniles perch as there were no differences in terms of specific growth rate between the treatments, but adults undergoing maturation should have at least partially LC‐PUFA in their diet, particularly arachidonic acid (ARA) which is important during maturation, as breeders are not able to bioconvert 18:2 n‐6 into ARA.     

Evaluation of Streptocephalus dichotomus nauplii as a larval diet for freshwater prawn Macrobrachium rosenbergii

This study was performed to assess the nutritional value of Streptocephalus dichotomus nauplii and compared with standard larval diet and parthenogenetic strain of Artemia nauplii. The effectiveness of live feed was determined by feeding freshwater prawn Macrobrachium rosenbergii postlarvae. Results on the growth rate, weight gain, survival, fatty acid and amino acid composition show a significant variation between the dietary treatments. Mean larval growth was significantly different ( P  < 0.001) between control diet (13.5 ± 0.5 mm) and live feeds ( Streptocephalus nauplii; 18.4 ± 0.5 mm and Artemia nauplii 18.7 ± 0.2 mm). The weight of larvae-fed Streptocephalus (41.1 ± 1.44 mg) and Artemia nauplii (41.7 ± 0.2 mg) was not significantly different; however, treatment with live feeds was significantly higher than the control (16.3 ± 0.5 mg) ( P  < 0.001). Proximate composition on the fairy shrimp reveals that they are rich in protein, lipid, essential amino acids (EAA) and essential fatty acids (EFA). The polyenoic unsaturated fatty acid (18 : 2 n-6 and 18 : 3 n-3) and highly unsaturated fatty acid (20 : 4 n-3 and 20 : 5 n-3) show a dramatic increase in larval tissue relative to its proportional composition in the live diets. Amino acid composition in the live feeds, Streptocephalus and Artemia nauplii, perfectly reflects on the amino acid proportions in the larvae-fed diets which confirm its utilization.