Sperm cryopreservation of Indian major carp, Labeo rohita: cryodiluents, sperm: cryodiluent dilution ratio and cryoprotectan t concentration

Cryogenic preservation trials of spermatozoa of Labeo rohita were carried out. Twenty four cryodiluents (extender + cryoprotectant), with the combination of six extenders such as egg-yolk citrate, urea-egg-yolk, 0.9% NaCl, Kurokura-2, Ma and Mb and four cryoprotectants viz. DMSO, glycerol, methanol and ethanol, were used to screen out the suitable cryodiluents. Sperm was preserved in 0.25ml plastic straw in programmable freezer. Two step freezing method was followed. Sperm preserved with egg-yolk citrate and urea-egg-yolk containing 10% DMSO showed best post-thaw motility (80%) followed by 0.9% NaCl (60%) and Kurokura-2(30%) solutions. Sperm with the extenders M" and Mb clotted at the time of equilibration and also after few days of preservation. Egg-yolk citrate mixed with ethanol and methanol also showed good percentage of motility (80%) but egg-yolk citrate with glycerol showed less sperm motility (>60%). To determine suitable dilution ratio of milt and cryodiluent two best extender eggyolk citrate and urea-egg-yolk with four cryoprotectants such as DMSO, glycerol, methanol and ethanol at different ratio viz 1:2,1:4,1:7,1:10,1:15 and 1:20 were used. Highest post-thaw motility (>80%) was observed when milt was preserved with egg-yolk citrate containing 10% DMSO at 1:2, 1:4, 1:7 and 1:10 dilutions. Meanwhile using glycerol as cryoprotectants provided less post thaw motility at lower dilution ratio but with the increase of its dilution showed good sperm motility compared with other cryoprotectants. Finally, evaluation on the effect of cryoprotectant concentration on post-thaw sperm motility was conducted. Egg-yolk citrate and four cryoprotectant i.e. DMSO, glycerol, methanol and ethanol with six different concentrations namely 5%,7%, 10%, 15%, 20% and 30%.were evaluated. Among the cryoprotectants DMSO, methanol and ethanol showed highest post-thaw motility (about 80%) at 7% and 10% concentrations. Although glycerol was not suitable at low concentration but its 20% and 30% concentration levels provided best post-thaw motility. No post-thaw motility was obtained with DMSO at 30% concentration. The overall analysis on cryoprotectant concentration indicated that below 5% and above 20% cryoprotectant concentrations could not be suitable for effective cryopreservation of spermatozoa.

Control of Variable Watermilfoil in Bashan Lake, CT with 2,4-D: Monitoring of Lake and Well Water.

Variable watermilfoil (Myriophyllum heterophyllum Michx.) has recently become a problem in Bashan Lake, East Haddam, CT, USA. By 1998, approximately 4 ha of the 110 ha lake was covered with variable watermilfoil. In 1999, the milfoil was spot treated with Aquacide®, an 18% active ingredient of the sodium salt of 2,4-D [(2,4-dichlorophenoxy) acetic acid], applied at a rate of 114 kg/ha. Aquacide® was used because labeling regarding domestic water intakes and irrigation limitations prevented the use of Navigate® or AquaKleen®, a 19% active ingredient of the butoxyethyl ester of 2,4-D. Variable watermilfoil was partially controlled in shallow protected coves but little control occurred in deeper more exposed locations. 2,4-D levels in the treatment sites were lower than desired and offsite dilution was rapid. In 2000, the United States Environmental Protection Agency (USEPA) issued a special local need (SLN) registration to allow the use of Navigate ® or AquaKleen® in lakes with potable and irrigation water intakes. Navigate® was applied at a rate of 227 kg/ha to the same areas as treated in 1999. An additional 2 ha of variable watermilfoil was treated with Navigate® in 2001, and 0.4 ha was treated in mid-September. Dilution of the 2,4-D ester formulation to untreated areas was slower than with the salt formulation. Concentrations of 2,4-D exceeded 1000 μg/ L in several lake water samples in 2000 but not 2001. Nearly all of the treated variable watermilfoil was controlled in both years. The mid-September treatment appeared as effective as the spring and early summer treatments. Testing of homeowner wells in all 3 years found no detectable levels of 2,4-D.(PDF contains 8 pages.)

Nutrient dynamics of seasonal tanks in the dry zone of Sri Lanka in relation to their hydrological regimes

Village tanks are put to a wide range of uses by the rural communities that depend on them for their survival. As the primacy of irrigation has decreased under these tanks due to a variety of climatic and economic reasons there is a need to reevaluate their use for other productive functions. The research presented in this paper is part of a programme investigating the potential to improve the management of living aquatic resources in order to bring benefits to the most marginal groups identified in upper watershed areas. Based on an improved typology of seasonal tanks, the seasonal changes and dynamics of various water quality parameters indicative of nutrient status and fisheries carrying capacity are compared over a period of one year. Indicators of Net (Primary) Productivity (NP): Rates of Dissolved Oxygen (DO) change, Total Suspended Solids (TSS): Total Suspended Volatile solids (TVSS) ratios are the parameters of principle interest. Based on these results a comparative analysis is made on two classes of ‘seasonal’ and ‘semi-seasonal’ tanks. Results indicate a broad correlation in each of these parameters with seasonal trends in tank hydrology. Highest productivity levels are associated with periods of declining water storage, whilst the lowest levels are associated with the periods of maximum water storage shortly after the NW monsoon. This variation is primarily attributed to dilution effects associated with depth and storage area. During the yala period, encroachment of the surface layer by several species of aquatic macrophyte also has progressively negative impacts on productivity. The most seasonal tanks show wider extremes in seasonal nutrient dynamics, overall, with less favourable conditions than the ‘semi-seasonal’ tanks. Never the less all the tanks can be considered as being highly productive with NP levels comparable to fertilised pond systems for much of the year. This indicates that nutrient status is not likely to be amongst the most important constraints to enhancing fish production. Other potential management improvements based on these results are discussed. [PDF contains 19 pages]

Einfluss der Verarbeitung auf lipophile chlororganische Rückstände in Fischen

Influence of fish processing methods on changes of chlordane and toxaphene residue concentrations were studied in various fishery products. Investigation included smoking, marinating and canning of fish. Smoking of Greenland halibut and ocean perch led to an increase in chlordane and toxaphene residue amounts in the edible part in relation to the raw material. A similar increase was observed during the marinating process of herring fillets. In both cases, loss of water was responsible for the observed changes in residue contents. The fat and thus the lipophilic organochlorine compounds remained completely in the products. Canning of herring fillets in tomato sauce reduced the residue concentration of the final product due to dilution by the tomato sauce. The concentrations in the fillets kept nearly unchanged even after storage of cans for six months. No equilibration was observed between residue concentrations in fillets and sauce.

Preliminary study on the natural extenders for artificial breeding of African catfish Clarias gariepinus (Burchell, 1822)

The objective of the present study was to determine the most suitable extender and their respective dilution ratios for African catfish sperm for artificial induced breeding and cryopreservation purposes. Three natural extenders were tested i.e. coconut water, sugarcane water and soybean solutions, at three different levels of sperm to extender dilutions of 1:20, 1:30 and 1:40. While Ringer solution was used as a control Diluted sperm were fertilized with ready isolated eggs to assess the fertility and hatching rate at 0, 6 and 12 hour intervals. The results showed that the eggs hatched approximately 19 to 27 hours after fertilization. In general, the fertilization and hatching rates decreased with increasing dilution ratio. With respect to natural extenders, the coconut water showed the highest fertility and hatching rates at 1:20 dilution ratio. Therefore, coconut water at 1:20 dilution ratio was the optimal condition for African catfish spermatozoa among the natural extenders investigated.

Report on the physical, bacteriological and biochemical analyses of seawater samples collected off existing sewage outfalls in the Sierra Leone River Estuary

The studies reported were undertaken as part of a wide environmental feasibility study for the establishment of a modern sewage system in Freetown. The aim of this part of the study was to determine whether the hydrological regime of the Sierra Leone River Estuary would permit the large-scale introduction of sewage into the estuary without damaging the environment. The important factors were whether: 1) there would be sufficient dilution of the sewage; 2) fleatable particles or other substances would create significant adverse effects in the estuarine ecosystem. The outfall sites are described together with the sampling stations, methods and analyses. Results include: 1) T/S profiles; 2) chemical analysis of the water. A review of literature on the Sierra Leone River Estuary is included which provides information on the plankton, benthos and fisheries. Results suggest that at certain points where local circulations occur it would be inadvisable to locate untreated sewage outfalls. Such points are frequently observed in small embayments. These studies have been of short duration but the data can serve as baseline for more extended investigations which would give a more complete picture of the seasonal patterns in the estuary.

Determination of dissolved oxygen in water with leukoberbelin-blue I. A quick Winkler method. [Translation from: Z.analyt.Chem. 262 97-99, 1972.]

The reaction of Mn(II) with water-dissolved oxygen, to a higher manganese hydroxide in an alkaline medium, as with the longstanding classic Winkler method, is the first step in the method described here. The assumption for faultless results by the conventional and modified Winkler method is clean water, which contains no organic substances by Mn(III) or Mn(IV). In many cases, however, eg. in river and lake-water tests, it can be seen with the naked eye that after some time the originally brown-coloured precipitate of manganese hydroxide becomes more and more colourless. Oxygen content was analysed in the water samples and evaluated by raising the amount of the leuko-base and giving the corresponding dilution of the colouring matter solution formed still higher oxygen contents can be measured.

Cypermethrin use in forestry and its impact on Welsh streams: An evaluation of the impact of cypermethrin use in forestry on Welsh streams

This is the Evaluation of the impact of cypermethrin use in forestry on Welsh streams from the University of Plymouth, published on September 2010 by the Environment Agency South West. The report focuses attention on Cypermethrin, a highly active synthetic pyrethroid insecticide effective against a wide range of pests in agriculture, public health, and animal husbandry. It is also used in forestry to control the pine weevil, Hylobius abietis. Cypermethrin is very toxic to aquatic invertebrates and fish at nanogram per litre concentrations. This project checks the effectiveness of current best practice measures in minimising the risk of pollution associated with the use of cypermethrin in forestry in Wales. Chemical results from the intensive studies show that cypermethrin entered minor watercourses draining treated areas at two of the eight sites. In one of these cases the level was well in excess of the short-term Predicted No Effect Concentration. The absence of a buffer area at the other site resulted in the cypermethrin reaching a main drain. However dilution appeared to be sufficient to prevent any impact on water quality or on the invertebrate community in the main stream. Invertebrate and chemical data from the extensive survey showed little evidence of pollution due to wider use of cypermethrin in Welsh forestry. Finally, a number of recommendations are made for further tightening controls on forestry practice to minimise the risk of cypermethrin entering the aquatic environment.

Studies on the status of pollution in certain estuarine areas in Kakinada

The BOD levels and saprotrophication of 2 stations in an estuarine habitat at Kakinada India, was studied during one full tidal cycle. Both the stations were found to be heavily polluted by sewage with BOD values ranging between 10.8 and 12.8 ppm at the commercial canal (station 1) and between 4.8 and 8.8 ppm at the mangrove creek (station 2). Tides did not seem to have much influence on the dilution of the pollutants. Based on planktonic indicator species observed and saprobity index which worked out to be 2.75, both the sampling areas are considered as heavily polluted alpha-mesosaprobic zones.

Distribution of surface sediments off Indus delta on the continental shelf of Pakistan

Surface sediments from the continental shelf area off Indus delta were analysed for their textural characteristics and carbonate content. The sediments are largely silt, silty clay and clayey silty sand. Sandy fraction is dominant in the outer region with relatively high carbonate content. The study shows that distribution of carbonate in sediments off Indus delta continental shelf is controlled by the dilution of terrigenous material and its distance from source area.

Cryopreservation of sperm of common carp, Cyprinus carpio and silver barb, Barbonymus gonionotus for genetically improved seed production

Experiments were conducted to develop and standardize the protocols for cryopreservation of sperm of common carp, Cyprinus carpio and also for using the cryopreserved sperm for fertilization of eggs. Nine extender solutions as Alsever's solution, kurokura-1, kurokura-2, urea egg-yolk, egg-yolk citrate, 0.6% glucose, 0.9% NaCl, Ma and Mb, and five cryoprotectants namely ethanol, methanol, dimethylsulfoxide (DMSO), dimethylamine (DMA) and glycerol were tested. The cryoprotectants were mixed at 10% concentration of the extenders (v/v) to make the cryodiluents. Milt and cryodiluents were mixed at a ratio of 1:9 for Alsever's solution, kurokura-1, kurokura-2, 0.6% glucose and 0.9% NaCl, 1:4 for urea egg-yolk, egg-yolk citrate, Ma and Mb. Among the cryodiluents Alsever's solution mixed with either ethanol or methanol was found to be suitable and it produced more than 90% and 80% spermatozoan motility at equilibrium and post-thaw periods, respectively. Kurokura-1 and kurokura-2 when mixed with the same cryoprotectants showed good spermatozoan motility at equilibrium period (80-90%) but the motility was reduced (30-55%) at post-thaw state. Other extenders did not produce acceptable sperm-motility and in some cases the frozen milt became clotted. Different dilution ratios (1:1, 1:2, 1:4, 1:5, 1:7, 1:9, 1:12, 1:15, 1:20) were formulated for obtaining a suitable milt dilution, the dilution ratio of 1: 9 (milt : cryodiluent) demonstrated the highest post-thaw spermatozoan motility (80%) in Alserver's solution. The optimum concentration of cryoprotectants in the cryodiluents was determined, 10% concentration level was found to be effective to produce the highest number of spermatozoan motility in comparison to the other concentrations (5%, 15%, 20% 30%). Sperm preserved with the cryodiluent Alsever's solution along with either methanol or ethanol was found to be effective to fertilize eggs and produce hatchlings. The hatching rates ranged between 1.48% and 14.76%, compare to control. The fish produced through use of cryopreserved sperm and normal sperm were found to grow well and no significant (P<0.05) growth difference was observed between them. In case of silver barb, Barbonymus gonionotus, sperm tested against six extenders such as egg-yolk citrate, urea-egg-yolk, kurokura-1, kurokura-2, 0.9% NaCl and modified fish ringer (MFR) solution. Cryoprotectants used were the same as those of C. carpio. Milt was diluted with the cryodiluent at a ratio of 1:4 for egg-yolk citrate and urea-egg-yolk, 1:5 for kurokura-1 and 1:9 for 0.9% NaCl, MFR and kurokura-2. The cryoprotectant concentration was maintained at 10% of the extender (v/v) in all the cases. Among the extenders, egg-yolk citrate and urea-egg-yolk mixed with 10% DMSO, methanol and ethanol produced 50% post-thaw spermatozoan motility, whereas DMA and glycerol provided only 10% motility. Trials on milt dilution ratio and cryoprotectant concentration are being conducted. Fertilization trials are also underway.