Embryonic and genetic manipulation in fish

Fishes, the biggest and most diverse community in vertebrates are good experimental models for studies of cell and developmental biology by many favorable characteristics. Nuclear transplantation in fish has been thoroughly studied in China since 1960s. Fish nuclei of embryonic cells from different genera were transplanted into enucleated eggs generating nucleo-cytoplasmic hybrids of adults. Most importantly, nuclei of cultured goldfish kidney cells had been reprogrammed in enucleated eggs to support embryogenesis and ontogenesis of a fertile fish. This was the first case of cloned fish with somatic cells. Based on the technique of microinjection, recombinant MThGH gene has been transferred into fish eggs and the first batch of transgenic fish were produced in 1984. The behavior of foreign gene was characterized and the onset of the foreign gene replication occurred between the blastula to gastrula stages and random integration mainly occurred at later stages of embryogenesis. This eventually led to the transgenic mosaicism. The MThGH-transferred common carp enhanced growth rate by 2-4 times in the founder juveniles and doubled the body weight in the adults. The transgenic common carp were more efficient in utilizing dietary protein than the controls. An "all-fish" gene construct CAgcGH has been made by splicing the common carp beta-actin gene (CA) promoter onto the grass carp growth hormone gene (gcGH) coding sequence. The CAgcGH-transferred Yellow River Carp have also shown significantly fast-growth trait. Combination of techniques of fish cell culture, gene transformation with cultured cells and nuclear transplantation should be able to generate homogeneous strain of valuable transgenic fish to fulfil human requirement in 21(st) century.

Estradiol stimulates growth hormone production in female goldfish

The effects of estradiol (E(2)) on growth hormone (GH) production was investigated in gonad-intact female goldfish. It was first necessary to generate a specific antibody for use in immunocytochemistry, Western, and dot-blot analyses of GH production. To accomplish this, grass carp GH (gcGH) cDNA was cloned by the reverse transcription polymerase chain reaction (RT-PCR) and expressed in Echerichia coli and a specific polyclonal antibody to recombinant gcGH was generated in the rabbit. In Western blot, the anti-gcGH antibody specifically immunoreacted with recombinant gcGH, purified natural common carp GH, and with a single 21.5-kDa GH form from pituitary extracts of grass carp, common carp, goldfish, and zebrafish but not salmon, trout, or tilapia. Intraperitoneal injection of the recombinant gcGH enhanced the growth rates of juvenile common carp demonstrating biological activity of this GH preparation. Electron microscopic studies showed that the anti-gcGH-I antibody specifically reacted with GH localized in the secretory granules of the goldfish somatotroph. Using anti-gcGH-I in a dot-blot assay, it was found that in vivo implantation of solid silastic pellets containing E(2) (100 mu g/g body weight for 5 days) increased pituitary GH content by 150% in female goldfish. In a second, independent study employing a previously characterized anticommon carp GH antibody for radioimmunoassay, it was found that E(2) increased pituitary GH content by 170% and serum GH levels by approximately 350%. The E(2)-induced hypersecretion of GH and increase in pituitary GH levels was not associated with changes in steady-state pituitary GH mRNA levels, suggesting that this sex steroid may enhance GH synthesis at the posttranscriptional or translational level. Previous observations indicate that GH can stimulate ovarian E(2) production. The present results show that E(2) can in turn stimulate GH production, indicating the existence of a novel pituitary GH-ovarian feedback system in goldfish. (C) 1997 Academic Press.

The culture-based fisheries in small, farmer-managed reservoirs in two provinces of northern Vietnam: an evaluation based on three production cycles

In this paper the results of a study conducted on the culture-based fisheries in small (ranging from 2 to 160 ha), farmer-managed reservoirs in YenBai and ThaiNguyen Provinces in the northern highland region of Vietnam, for the production cycles of 1997/98, 98/99 and 99/00 are presented. The small reservoirs are leased to small farmers by the provincial authorities for fishery activities, and all lessees adopt culture-based fisheries when fingerlings of grass carp (Ctenopharyngodon idella), silver carp (Hypophthalmichthys molitrix), bighead carp (Aristichthys nobilis), common carp (Cyprinus carpio) and mrigal (Cirrihinus mrigala) are stocked between March and mid-April each year and harvested, using large seine nets, after approximately 11-12 months. The mean yields from reservoirs in YenBai and ThaiNguyen Provinces in 97/98, 98/99 and 99/00 production cycles were 251, 332 and 253, and 331, 372 and 210 kg ha⁻¹ respectively. There were major differences in the fish productivity in the reservoirs in the two Provinces, and in a reservoir between culture cycles. The stocking strategies appeared to be rather ad hoc, being determined by the availability of seed stock and the financial status of the lessees. Accordingly, there was no apparent consistent trend in the improvement of yields from the culture-based fishery practice throughout the growth cycles. The fish yields in reservoirs in each Province were significantly related to reservoir area (exponentially) and to mean weight of stocked fish and conductivity (logarithmically). Of the stocked fish, the highest returns were obtained with mrigal and bighead carp, which collectively contributed > 50% to the harvest. The return from common carp was the lowest. The mean growth rate of grass carp (2.7 g day⁻¹), followed by bighead carp (2.0 g day⁻¹) was the highest in reservoirs in YenBai Province, bighead carp (4.0 g day⁻¹) followed by grass carp (3.2 g day⁻¹) was the highest in ThaiNguyen Province. The seed stocked on average accounted for 65% and 48% of the total operating costs in YenBai and ThaiNguyen Provinces, and the mean cost:benefit ratio of the culture-based fishery in the two Provinces was 0.35 and 0.37 respectively. The culture-based fishery on average contributed about 28% to the gross income of a farmer lessee.

Culture-based fisheries in small reservoirs in northern Vietnam: effect of stocking densities and species combinations

This study was conducted in 20 reservoirs, ranging in size from 4 to 30 ha, in the mountainous, northern region of Vietnam, in ThaiNguyen and YenBai provinces, over two growth cycles in 2002/03 and 2003/04. The reservoirs are leased by farmers for fishery activities from the provincial administration, and the trials were managed by the lessee farmers. Three species combinations in ratios (by fingerling weight) of grass carp: silver carp: bighead carp: common carp: mrigal 1:2:1:1:3 (A), 1:3:1:1:2 (B) and 1:2:1:1:2 (C) were used as seed stock. The overall mean yield of stocked fish in 2002/03 and 2003/04 growth cycles in reservoirs in ThaiNguyen and YenBai provinces was 165 (±21) and 190 (39), and 287 (±22) and 325 (±24) kg ha⁻¹ respectively. The yield in reservoirs in both provinces, in both growth cycles and irrespective of the species combinations, increased in relation to stocking density. In reservoirs in ThaiNguyen Province, the species combination B gave the lowest yield (both growth cycles and overall), and differed significantly (P<0.05) from combinations A and C. The stocking efficiency (ratio of the yield of stocked fish in kg ha⁻¹ to the weight of the stocked fish in kg ha⁻¹) in reservoirs in ThaiNguyen Province ranged from 2.9 to 5.1 over the two growth cycles and that in YenBai from 2.8 to 3.9. There was no discernible trend between growth cycles and/or between species combinations. The major cost incurred was for fingerling procurement. In all instances, a net profit was accrued. The mean (±standard error (SE)) net profit ha⁻¹ (in 103VN dong; approximately 15 500 VND=1 US$) was 885 (±270) and 864 (±214), and 1322 (±176) and 1600 (±150) for the growth cycles 2002/03 and 2003/04 for reservoirs in ThaiNguyen and YenBai provinces respectively. Between the two growth cycles, the net profit increased in eight and three reservoirs from YenBai and ThaiNguyen, respectively, the maximum increase being recorded in Khuan Gio (165%) and Dong Man (39%) reservoirs.

Policultivos de peixes integrados à produção vegetal: avaliação econômica e sócio ambiental (Peixe-verde)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Preferência alimentar e eficiência da carpa capim (Ctenopharyngodon idella) no controle de macrófitas aquáticas em mesocosmos

Pós-graduação em Agronomia (Produção Vegetal) - FCAV

Características tecnológicas de espécies de pescado com potencial de criação na região sudoeste do Paraná

The fish meat has a particular chemical composition which gives its high nutritional value. However, this food is identified for being highly perishable and this aspect is often named as a barrier to fish consumption. The southwestern Paraná region, parallel to the country's reality, it is characterized by low fish consumption; and one of the strategies aimed at increasing the consumption of this important protein source is encouraging the production of other species besides tilapia. Within this context, it is necessary to know about the meat characteristics. In this sense, the objective of this study was to evaluate the technological potential of pacu, grass carp and catfish species. To do so, at first, it was discussed the chemical and biometric assessment under two distinct descriptive statistical methods, of the three species; and it was also evaluated the discriminating capacity of the study. In a second moment, an evaluation of effects done by two different processes of washing (acid and alkaline) regarding the removal of nitrogen compounds, pigments and the emulsifying ability of the proteins contained in the protein base obtained. Finally, in the third phase, it was aimed to realize the methodology optimization in GC-MS for the analysis geosmin and MIB (2-metilisoborneol) compounds that are responsible for taste/smell of soil and mold in freshwater fish. The results showed a high protein and low lipid content for the three species. The comparison between means and medians revealed symmetry only for protein values and biometric measurements. Lipids, when evaluated only by the means, overestimate the levels for all species. Correlations between body measurements and fillet yield had low correlation, regardless of the species analyzed, and the best prediction equation relates the total weight and fillet weight. The biometric variables were the best discriminating among the species. The evaluation of the washings, it was found that the acidic and basic processes were equally (p ≥ 0.05) efficient (p ≤ 0.05) for the removal of nitrogen compounds on the fish pulps. Regarding the extraction of pigments, a removal efficiency was recorded only for the pacu species, the data were assessed by the parameters L *, a *, b *. When evaluated by the total color difference (ΔE) before and after washing for both processes (acid/alkaline) the ΔE proved feasible perceived by naked eye for all species. The catfish was characterized as the fish that presents the clearest meat with the basic washing considered the most effective in removing pigments for this species. Protein bases obtained by alkaline washes have higher emulsifying capacity (p ≤ 0.05) when compared to unwashed and washed in acid process pulps. The methodology applied for the quantification of MIB and geosmin, allowed to establish that the method of extraction and purification of analytes had low recovery and future studies should be developed for identification and quantification of MIB and geosmin on fish samples.

Role of Aeromonas hydrophila in bacterial septicemia of cultured carps in Khouzestan Province and investigation on protectivity of bacterin (s) prepared from acute isolate(s) of it

Motile Aeromonas are the most common bacteria of freshwater in the world that cause disease in fish and other cold-blooded and warm-blooded hosts. Among this group of bacteria, Aeromonas hydrophila is important in causing complications such as fin rot, skin ulcers and lethal hemorrhagic septicemia in fish. Several virulence factors involved in the pathogenesis of Aeromonas hydrophila, including extracellular enzymes (protease, lipase, elastase, gelatinase and nuclease) and toxins. From the exotoxins, hemolysin, aerolysin and cytolytic enterotoxin play an important role in pathogenesis. Detection of virulence markers by PCR as a key component of determining the pathogenesis of the bacteria and using indigenous vaccines for better immunization against this disease is important. In this study, a total of 200 fanned carps (126 common carp. 39 silver carp and 35 of grass carp) with symptoms suspected aeromonas septicemia were isolated from Khouzestan province farms. 125 bacteria belong to Aeromonas genus detected by biochemical and PCR methods. 31 of all isolates recognized as Aeromonas hydrophila with biochemical methods, I6srRNA detection and Lipase genes. Results showed that the role of Aeromonas sp. and Aeromonas hydrophila in fish with disease symptoms were 62.5% and 15.5% respectively. By using specific primers, three virulence genes including hemolysin, aerolysine and cytolytic enterotoxin were detected in these confirmed isolates, that 18 isolates (58/06%) hemolysin positive (hlyA +), 16 isolates (51/61%) aerolysine positive (aerA+) and 23 isolates (74/19%) for cytolytic enterotoxin gene (act+) were positive. The result of present study showed that most of the confirmed isolates genotype was hlyA+ act- with frequency equal to 51/61%. For investigating the protection effect of acut strain of bacteria, UV inactivated bacterin was used.

Designing for participation in local social ridesharing networks : grass roots prototyping of IT systems

This abstract explores the possibility of a grass roots approach to engaging people in community change initiatives by designing simple interactive exploratory prototypes for use by communities over time that support shared action. The prototype is gradually evolved in response to community use, fragments of data gathered through the prototype, and participant feedback with the goal of building participation in community change initiatives. A case study of a system to support ridesharing is discussed. The approach is compared and contrasted to a traditional IT systems procurement approach.

Plastic traits of an exotic grass contribute to its abundance but are not always favourable

In herbaceous ecosystems worldwide, biodiversity has been negatively impacted by changed grazing regimes and nutrient enrichment. Altered disturbance regimes are thought to favour invasive species that have a high phenotypic plasticity, although most studies measure plasticity under controlled conditions in the greenhouse and then assume plasticity is an advantage in the field. Here, we compare trait plasticity between three co-occurring, C 4 perennial grass species, an invader Eragrostis curvula, and natives Eragrostis sororia and Aristida personata to grazing and fertilizer in a three-year field trial. We measured abundances and several leaf traits known to correlate with strategies used by plants to fix carbon and acquire resources, i.e. specific leaf area (SLA), leaf dry matter content (LDMC), leaf nutrient concentrations (N, C:N, P), assimilation rates (Amax) and photosynthetic nitrogen use efficiency (PNUE). In the control treatment (grazed only), trait values for SLA, leaf C:N ratios, Amax and PNUE differed significantly between the three grass species. When trait values were compared across treatments, E. curvula showed higher trait plasticity than the native grasses, and this correlated with an increase in abundance across all but the grazed/fertilized treatment. The native grasses showed little trait plasticity in response to the treatments. Aristida personata decreased significantly in the treatments where E. curvula increased, and E. sororia abundance increased possibly due to increased rainfall and not in response to treatments or invader abundance. Overall, we found that plasticity did not favour an increase in abundance of E. curvula under the grazed/fertilized treatment likely because leaf nutrient contents increased and subsequently its' palatability to consumers. E. curvula also displayed a higher resource use efficiency than the native grasses. These findings suggest resource conditions and disturbance regimes can be manipulated to disadvantage the success of even plastic exotic species.

An invasive grass shows colonization advantages over native grasses under conditions of low resource availability

Increased or fluctuating resources may facilitate opportunities for invasive exotic plants to dominate. This hypothesis does not, however, explain how invasive species succeed in regions characterized by low resource conditions or how these species persist in the lulls between high resource periods. We compare the growth of three co-occurring C4 perennial bunchgrasses under low resource conditions: an exotic grass, Eragrostis curvula (African lovegrass) and two native grasses, Themeda triandra and Eragrostis sororia. We grew each species over 12 weeks under low nutrients and three low water regimes differentiated by timing: continuous, pulsed, and mixed treatments (switched from continuous to pulsed and back to continuous). Over time, we measured germination rates, time to germination (first and second generations), height, root biomass, vegetative biomass, and reproductive biomass. Contrary to our expectations that the pulsed watering regime would favor the invader, water-supply treatments had little significant effect on plant growth. We did find inherent advantages in a suite of early colonization traits that likely favor African lovegrass over the natives including faster germination speed, earlier flowering times, faster growth rates and from 2 weeks onward it was taller. African lovegrass also showed similar growth allocation strategies to the native grasses in terms of biomass levels belowground, but produced more vegetative biomass than kangaroo grass. Overall our results suggest that even under low resource conditions invasive plant species like African lovegrass can grow similarly to native grasses, and for some key colonization traits, like germination rate, perform better than natives.

Unravelling the mechanisms behind the invasion of an introduced and now undesirable grass species

The introduction of Eragrostis curvula (African Lovegrass, herafter Lovegrass) for pasture improvement across Australia has not been successful. Instead Lovegrass, a C4 perennial grass originating from Southern African, has proven unpalatable to stock and to have low nutritional value if stocks do eat it. It has spread prolifically along roadsides, stream banks, conservation areas and pastures. Because control efforts have not been effective, our aim was to determine the putative mechanisms responsible for the dominance of Lovegrass, specifically disturbance (selective grazing) and competition.