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.

Concentration and distribution of heavy metals (lead, cadmium, chromium, copper, nickel, zinc) in Karachi shore and offshore sediments

A study to measure the heavy metal pollution level in the sediment of coastal and offshore area indicates that high concentration of heavy metals were found around Manora channel and eastern coast of Karachi. In comparison with coastal areas, relatively low concentration of heavy metals was recorded in the offshore area. The result shows that sewage and industrial wastes are the main source of heavy metal pollution in the coastal area. The concentration of heavy metals in the sediments is as follows: Chromium 10.4-33.69, nickel 13.3-47.6, lead 10.0-39.04, cadmium 0.08-0.21, zinc 7.4-73.2 and copper 9.44-18.56 mg/kg. In the offshore areas strong correlation was observed between copper and organic carbon, and calcium carbonate and cadmium. In the shore area such correlation has been recorded among nickel, chromium, zinc, and chromium and copper. The Karachi. coast is viewed as moderately polluted when compared to other continental coastal areas.

Effect of sublethal concentration of artificial seawater on oxygen uptake and opercular frequency in an Indian major carp, Labeo rohita (Ham.)

The present study reports the effect of artificial seawater on oxygen uptake and opercular frequency in an Indian major carp, Labeo rohita. Whereas a control fish of 7.34 g average body weight consumed 1.538 ml O sub(2.) hˉ¹, the 24h and 96h exposed fish of the same body weight consumed 1.07 4 and 0.897 ml O sub(2.) hˉ¹, respectively. The oxygen uptake per unit body weight under controlled condition was 0.219 ml. gˉ¹. hˉ¹, whereas in 24h and 96h exposed fish, it was 0.152 and 0.124 ml. gˉ¹, hˉ¹, respectively. The change in opercular movement in 24h exposed fish was 7.67% higher, whereas in 96h exposed fish, it was 22.43% higher as compared to the control one. All changes are highly significant (p<0.001).

Immnunomodulatory and cytotoxic effects of aqueous leaf extracts of certain plants on common carp, Cyprinus carpio

Effect of aqueous leaf extracts of Catheranthus roseus, Calotropis gigantium and Datura stromoneum on common carp, Cyprinus carpio were investigated. C. carpio were separately fed with 1 and 2% aqueous extracts of these three plant leaves for a period of seven days. In 1% Catheranthus roseus of leaf extract fed group no significant tissue level changes were recorded. One and 2% of other two species fed treated group showed mild to severe necrotic and cellular changes in liver, kidney and spleen. Immunologically, significant rise in antibody titre and respiratory burst activity was recorded for 1% Catheranthus roseusfed group.

Heavy metal concentration in South Vietnam waters

Data obtained from investigations which had been carried out during 1998-1999 in the waters of South Vietnam indicate that concentrations of Fe and Zn were usually higher than permissible limits. In the East South Vietnam waters, Fe concentration varied from 77 to 4450g/l (mean 1045g/l) and Zn concentration varied from 7.24 to 74.1g/l (mean 26.7g/l), Pollution coefficients of the two metals were 10.4 and 2.7 respectively. In the Gulf of Thailand, variation range of Fe was 133-850g/l (mean 318g/l) and of Zn was 4.8-167.8g/l (mean 23.46g/l); corresponding pollution coefficients were 3.2 and 2.3. Mn, Cu, As had mean concentrations lower than permissible limits. In the East South Vietnam waters, variation range for Mn was 9.0-444.6g/l (mean 57.0g/l), for Cu was 1.0- 20.3g/l (mean 3.1g/l) and for As was 1.0-10.5 g/l (mean 3.7 g/l). In the Gulf of Thailand, Mn concentration varied from 1.2 to 410.5g/l (mean 19.1g/l), Cu concentration varied from 1.2 to 14.7g/l (mean 3.7g/l), variation range of As concentration was 1.0-13.3g/l with the mean value was 3.6g/l. Scarce data relating to Hg, Cd, Pb showed that their concentrations in the East South Vietnam waters were negligible (usually < 1.0g/l).

Preservation of cotton fish net twines by tanning 1. Optimum concentration of tanning

Samples of tannin-containing preservatives used by fishermen in India for treating cotton nets were collected and qualitative and quantitative characterisation of the tannins made. The concentrations of different tannins required to impart optimum periods of preservation to the net were worked out and found to be 2% in 8 out of 10 materials studied.

On the measurement of concentration of blanching brine by electrical conductivity method

Blanching is an important operation in the shrimp canning process, in order to bring down the moisture content of the product to the required level, to allow the proteins to coagulate and to give proper texture, shape and characteristic pink colour to the meat. It has been observed that among other factors responsible for fluctuations in the drained weight in the canned prawns, concentration of brine used for blanching and the duration of blanching are important (Varma, Chaudhury and Pillai 1961). The papers gives details of experiments

A phylogeny of Chinese leaf monkeys using mitochondrial ND3-ND4 gene sequences

The phylogeny of Chinese leaf monkeys, especially the snub-nosed monkeys (Rhinopithecus), has not been thoroughly investigated using molecular sequence data, perhaps due to their rarity in the wild and their poor representation in institutional collections. Despite several proposed classifications, systematic relationships of these species remain poorly defined and this has hindered their conservation. To clarify the phylogenetic relationships of the leaf monkey clade in China, we sequenced the mitochondrial ND3, ND4L, ND4, tRNA(Arg), tRNA(His), tRNA(Ser), and tRNA(Leu) genes for Rhinopithecus bieti, R. roxellana, Trachypithecus francoisi, T. f. leucocephalus, and T. phayrei as well as Pygathrix nemaeus and Colobus guereza. We included a rotal of 2252 characters for each individual, excluding gaps in primary sequences. Our interpretation of the results from character- and distance-based phylogenetic analyses suggest that (1) Pygathrix nemaeus is sister to Rhinopithecus rather than to Trachypithecus though it is quite divergent from the former; (2) the Yunnan snub-nosed monkey, Rhinopithecus bieti, represents a valid species; (3) the white-headed leaf monkey is not a distinct species, but instead is a subspecies of Trachypithecus francoisi (T. f. leucocephalus), though it should still be considered a separate evolutionarily significant unit (ESU); and (4) because two individuals of the Phayrei's leaf monkey, T. phayrei, are genetically distinct from one another, a more extensive revision of the taxonomy of this putative species in China is needed. These results, plus ongoing work on the molecular systematics of the entire Asian leaf monkey radiation, can provide a sound basis for identifying the appropriate units of conservation for this endangered group of primates.

Adaptive evolution of a duplicated pancreatic ribonuclease gene in a leaf-eating monkey

Although the complete genome sequences of over 50 representative species have revealed the many duplicated genes in all three domains of life(1-4), the roles of gene duplication in organismal adaptation and biodiversity are poorly understood. In addition,

Pseudogenization of the tumor-growth promoter angiogenin in a leaf-eating monkey

Physiological functions of human genes may be studied by gene-knockout experiments in model organisms such as the mouse. This strategy relies on the existence of one-to-one gene orthology between the human and mouse. When lineage-specific gene duplication occurs and paralogous genes share a certain degree of functional redundancy, knockout mice may not provide accurate functional information on human genes. Angiogenin is a small protein that stimulates blood-vessel growth and promotes tumor development. Humans and related primates only have one angiogenin gene, while mice have three paralogous genes. This makes it difficult to generate angiogenin-knockout mice and even more difficult to interpret the genotype-phenotype relation from such animals should they be generated. We here show that in the douc langur (Pygathrix nemaeus), an Asian leaf-eating colobine monkey, the single-copy angiogenin gene has a one-nucleotide deletion in the sixth codon of the mature peptide, generating a premature stop codon. This nucleotide deletion is found in five unrelated individuals sequenced, and therefore is likely to have been fixed in the species. Five colobine species that are closely related to the douc langur have intact angiogenin genes, suggesting that the pseudogenization event was recent and unique to the douc langur lineage. This natural knockout experiment suggests that primate angiogenin is dispensable even in the wild. Further physiological studies of douc largurs may offer additional information on the role of this cancer-related gene in normal physiology of primates, including humans. Our findings also provide a strong case for the importance of evolutionary analysis in biomedical studies of gene functions. (C) 2003 Elsevier Science B.V. All rights reserved.

Antioxidant effect of betel leaf extract on dry-cured fish

The effect of betel leaf (Piper betle Linn.) extract on control of autoxidation of fat in dry fish has been studied. Oil sardine has been selected for experiments since it contains very high amount of fat. The treatments were given with 5% (w/v) betel leaf extract in water at different stages of salt curing. FFA, PV and TVN values of the samples were determined periodically to assess the keeping quality and autoxidation. The sample, prepared by dipping the fish in the extract immediately after salting and then drying as usual, was found to have better keeping qualities.

Phylogenetic incongruence between nuclear and mitochondrial markers in the Asian colobines and the evolution of the langurs and leaf monkeys

Evidence of incongruence between mitochondrial and nuclear gene trees is now becoming documented with increasing frequency. Among the Old World monkeys, this discordance has been well demonstrated in the Cercopithecinae, but has not yet been investigated

Adaptive Evolution of Digestive RNASE1 Genes in Leaf-Eating Monkeys Revisited: New Insights from Ten Additional Colobines

Pancreatic RNase genes implicated in the adaptation of the colobine monkeys to leaf eating have long intrigued evolutionary biologists since the identification of a duplicated RNASE1 gene with enhanced digestive efficiencies in Pygathrix nemaeus. The recent emergence of two contrasting hypotheses, that is, independent duplication and one-duplication event hypotheses, make it into focus again. Current understanding of Colobine RNASE1 gene evolution of colobine monkeys largely depends on the analyses of few colobine species. The present study with more intensive taxonomic and character sampling not only provides a clearer picture of Colobine RNASE1 gene evolution but also allows to have a more thorough understanding about the molecular basis underlying the adaptation of Colobinae to the unique leaf-feeding lifestyle. The present broader and detailed phylogenetic analyses yielded two important findings: 1) All trees based on the analyses of coding, noncoding, and both regions provided consistent evidence, indicating RNASE1 duplication occurred after Asian and African colobines speciation, that is, independent duplication hypothesis; 2) No obvious evidence of gene conversion in RNASE1 gene was found, favoring independent evolution of Colobine RNASE1 gene duplicates. The conclusion drawn from previous studies that gene conversion has played a significant role in the evolution of Colobine RNASE1 was not supported. Our selective constraint analyses also provided interesting insights, with significant evidence of positive selection detected on ancestor lineages leading to duplicated gene copies. The identification of a handful of new adaptive sites and amino acid changes that have not been characterized previously also provide a necessary foundation for further experimental investigations of RNASE1 functional evolution in Colobinae.

Aniracetam restores the effects of amyloid-beta protein or ageing on membrane fluidity and intracellular calcium concentration in mice synaptosomes

In the present study, we observed the in vitro effect of aniracetam on membrane fluidity and free calcium concentrations (Ca(2+)i) of frontal cortical (FC) and hippocampal (HP) synaptosomes of aged mice and young mice treated with amyloid-beta protein (A

Relationship between trichosanthin cytotoxicity and its intracellular concentration

Trichosanthin (TCS) is a type I ribosome-inactivating protein with board spectrum of biological activity. Toxicity of this compound differs in different cell lines and this study examined the cause of such difference. It is generally believed that TCS toxicity is mediated through intracellular ribosome inactivation. Therefore, TCS toxicity should be determined by the amount inside cells rather than outside. Three different cell types IC21, JAR and Vero cell lines were chosen with high, medium and low sensitivity to TCS. Intracellular concentrations of fluorescein isothiocyanate labeled TCS were determined by laser scanning confocal microscopy. A good relationship was demonstrated between intracellular TCS concentration and toxicity. Highest intracellular concentration was found in IC21, followed by JAR, and lowest in Vero cells. When the intracellular TCS concentrations in these cells were reduced by using a competitive inhibitor to block cell entry, cytotoxicity was not observed. In conclusion, there is strong evidence to indicate that cytotoxicity of TCS is dependent on its intracellular concentration. Variation of cytotoxicity in different cells may be related to the mechanisms affecting its internalization. (C) 2002 Published by Elsevier Science Ireland Ltd.