GENETIC DIVERSITY IN THE CHINESE PANGOLIN (MANIS-PENTADACTYLA) - INFERRED FROM RESTRICTION ENZYME ANALYSIS OF MITOCHONDRIAL DNAS

Two different forms of Chinese pangolins can be recognized according to the color of their scales, i.e., brown and dusky. We analyzed mitochondrial DNA (mtDNA) purified from the livers of seven dusky and six brown Chinese pangolins from the same locality, using cleavage patterns from 19 restriction enzymes. From the 19 6-bp recognition enzymes used, 51-56 sites were observed. By combining the cleavage patterns for each enzyme, the 13 samples were classified into four restriction types: two in dusky and two in brown Chinese pangolins. The estimated number of nucleotide substitutions per site in dusky and brown types is 0.002, and that between dusky and brown types is 0.012. Divergence between brown and dusky forms began 0.6 Myr ago, provided the mean rate of sequence divergence is 0.02 per Myr in mtDNA. Our results suggest that there is considerable divergence in Chinese pangolins, and brown and dusky Chinese pangolins may be quite different forms or, at least, belong to different maternal groups.

PHYLOGENY OF RHESUS-MONKEYS (MACACA-MULATTA) AS REVEALED BY MITOCHONDRIAL-DNA RESTRICTION ENZYME ANALYSIS

Mitochondrial DNA, purified from 36 samples of 23 local populations which are widely distributed in Vietnam, Burma, and 10 provinces of China, has been analyzed to model the phylogeny of rhesus monkeys. The 20 local populations of China may represent nearly all major populations in China. Using 20 restriction endonucleases of 6-bp recognition, we observed a total of 50-61 sites in the various samples. By combining the cleavage patterns for each enzyme, the 36 samples were classified into 23 restriction types, each of which was found exclusively in the respective population from which samples were obtained By combining the earlier study of Indian rhesus monkeys, phylogenetic trees, which have been constructed on the basis of genetic distance, indicate that rhesus monkeys in China, Vietnam, India, and Burma can be divided into seven groups. Integrating morphological and geographical data, we suggest that rhesus monkeys in China, Vietnam, and Burma may be classified into six subspecies-M. m. mulatta, M. m. brevicaudus, M. m. lasiotis, M. m. littoralis, M. m. vestita, and M. m. tcheliensis-and rhesus monkeys in India may be another valid subspecies. M. m. tcheliensis is the most endangered subspecies in China. Divergence among subspecies may have begun 0.9-1.6 Ma. The radiation of rhesus monkeys in China may have spread from the southwest toward the east. The taxonomic status of the Hainan monkey and the Taiwan monkey require further investigation.

PHYLOGENETIC-RELATIONSHIPS OF MACAQUES AS INFERRED FROM RESTRICTION-ENDONUCLEASE ANALYSIS OF MITOCHONDRIAL-DNA

Mitochondrial DNAs (mtDNAs) purified from 25 samples of 6 species of macaques, Macaca mulatta, M. fascicularis, M. arctoides, M. nemestrina, M. assamensis and M. thibetana, were analyzed to study the phyletic relationships among the species. A total of 36-46 sites was observed in each sample. By combining the cleavage patterns for each of the endonucleases, the 25 samples were classified into 11 restriction types. When data on M. fuscata and M. cyclopis collected by other authors were added to our own, the resultant molecular phylogenetic trees indicated that the 8 species may be divided into 4 groups: (1) M. mulatta, M. fuscata, M. cyclopis and M. fascicularis; (2) M. arctoides, (3) M. nemestrina; (4) M. assamensis and M. thibetana. Our results suggest that within both the fascicularis and sinica groups genetic distances are small between members and that the status of the species within the groups may require further investigation.

PHYLOGENY OF THE SLOW LORISES (GENUS NYCTICEBUS) - AN APPROACH USING MITOCHONDRIAL-DNA RESTRICTION ENZYME ANALYSIS

Mitochondrial DNA polymorphisms in 15 specimens of three species of slow lorises-Nycticebus coucang, N. intermedius, and N. pygmaeus-were analyzed in order to study the evolutionary relationships among the species. Eight restriction types were observed in the samples. Phylogenetic trees constructed on the basis of genetic distances showed that the slow lorises sort into two clusters: four types of N. coucang and three types of N. intermedius plus one type of N. pygmaeus. Our results suggest that there are two valid species in the genus Nycticebus-N. coucang and N. pygmaeus-and that N. intermedius should be included within N. pygmaeus. Divergence between the two species may have begun 2.7 Ma (million years ago). Evolution of gross morphology, chromosomes, and mitochondrial DNA in the slow lorises appears to be concordant.

Phylogenetic relationships of five pika species (genus Ochotona) based on mitochondrial DNA restriction maps

Restriction site mapping of mitochondrial DNA (mtDNA) with 16 restriction endonucleases was used to examine the phylogenetic relationships of Ochotona cansus, O. huangensis, O. thibetana, O. curzoniae and O. erythrotis. A 1-kb length variation between O. erythrotis of subgenus Pika and other four species of subgenus Ochotona was observed, which may be a useful genetic marker for identifying the two subgenera. The phylogenetic tree constructed using PAUP based on 61 phylogenetically informative sites suggests that O. erythrotis diverged first, followed by O. cansus, while O. curzoniae and O. huangensis are sister taxa related to O. thibetana, The results indicate that both O. cansus and O. huangensis should be treated as independent species. If the base substitution rate of pikas mtDNA was 2% per million years, then the divergence time of the two subgenera, Pika and Ochotana, is about 8.8 Ma ago of late Miocence, middle Bao-dian of Chinese mammalian age, and the divergence of the four species in subgenus Ochotona would have occurred about 2.5 - 4.2 Ma ago, Yushean of Chinese mammalian age. This calculation appears to be substantiated by the fossil record.

Effect of oxidation of dietary fish lipids on the quality of proteins in diet

The paper reports results of a study on the effect of oxidised fish oil in the diet on the quality of dietary fish proteins. With increased oxidation of oil, digestibility and utilization of the protein registered a decrease. Oxidation of fish oil beyond a stage was found to affect the intake of diet itself and rats lost weight rapidly.

Phylogeny of Muntiacus (Cervidae) based on mitochondrial DNA restriction maps

Mitochondrial DNA restriction maps for 12 restriction enzymes of four species of muntjacs-Indian muntjac (M. muntjak), Gongshan muntjac (M. gongshanensis), black muntjac (M. crinifrons), and Chinese muntjac (M. reevesi)-were compared to estimate the phylogenetic relationships among them. Phylogenetic trees were constructed by both distance and parsimony methods. The two resulting trees share a similar topology, which indicates that the black muntjac and the Gongshan muntjac are closely related, followed by the Chinese muntjac; the Indian muntjac is the sister taxon to all the other muntjacs.

Dietary crude protein requirement of Tilapia nilotica fry

To determine growth and survival of Tilapia nilotica fry fed formulated practical dry diets with varying crude protein levels, fish were subjected to 3 seperate feeding trials. Isocaloric practical diets containing 20, 25, 30 and 35% crude protein were fed to fry at 15% fish biomass daily for 7 weeks in 2 trials and another set containing 20, 25, 30, 35, 40, 45 and 50% was given for 8 weeks. On the basis of growth, survival and feed conversion, T. nilotica fry required 35% crude protein in the practical diets given at 15% fish biomass.

Influence of dietary proteins on cholesterol levels in albino rats

Six groups of albino rats were fed identical diets, differing in their protein sources for sixty days. The protein sources used were fat free casein, defatted groundnut cake, and defatted fish powders from three species of fishes, namely, the fresh water fish Labeo rohita, the marine fishes, Rastrelliger kanagurta and Otolithus argenteus and marine prawns, Parapenaeopsis stylifera. After sixty days, the levels of cholesterol in the serum, liver and heart of the rats were estimated. The casein group recorded the highest level of cholesterol. Compared to casein, the groundnut protein was distinctly hypocholesterolemic. The fish proteins had a still greater cholesterol lowering ability. Of the proteins used in this study, the proteins of prawns had the maximum hypocholesterolemic effect. An attempt is made to correlate the behaviour of the proteins in this respect, with their respective amino acid compositions.

Effects of dietary supplementation of commercial human probiotic and antibiotic on the growth rate and content of intestinal microflora in ornamental fishes

The effects of dietary supplementation of commercial human probiotic, Lactobacil and antibiotic, oxytetracycline on the growth, survival, disease resistance and content of intestinal microflora in two ornamental fishes, viz., goldfish, Carassius auratus and swordtail, Xiphophorus helleri were studied. The total wet weight gain, food conversion ratio and specific growth rate of C. auratus did not vary significantly (p>0.05) among treatments. While in X. helleri, significant differences existed in the total wet weight gain, survival, food conversion ratio and specific growth rate among treatment groups (p<0.05). The counts of antibiotic resistant bacteria in fish gut increased with days of culture in all the treatments and the increase was more in antibiotic fed fishes. A reduction in the development of antibiotic resistance among the bacterial flora of fish gut was noticed in probiotic fed groups of C auratus and X. helleri. The results of the present study revealed that the effects of human probiotic on the growth, survival and disease resistance of ornamental fishes are variable and difficult to reproduce the similar effect on different species.

Influences of dietary lipid and phosphorus levels on retention and excretion of phosphorus and nitrogen in fingerlings red sea bream, Pagrus major

A laboratory based 2x3 factorial experiment was conducted for 12 weeks to investigate the influences of dietary lipid and phosphorus (P) levels on retention and excretion of phosphorus and nitrogen (N) in fingerling red sea bream. Two levels of lipid (210 and 260 g/kg) and three levels of phosphorus (17, 14 and 12 g/kgˉ¹) in the dry diets were tested. Duplicate groups of 25 red sea bream (average weight 3.74±0.07 g) per 60L glass tank were fed experimental diets three times a day near to satiation level at 22 to 28°C water temperature. A reduction in dietary fish meal from 500 to 300 g/kg dry diet, corresponding to a supplementation in both dietary lipid and P resulted in significant increase in both P and N retention which resulted in the reduction of their excretion by red sea bream. The overall results of the present study demonstrated that both lipid and phosphorus supplementation are necessary for developing less-polluting feed which in turn, reduce fish meal level in the diet of fingerling red sea bream. Further studies in this regard with different size and age groups of red sea bream are warranted.

Necessity of dietary calcium supplement in file fish (Monacanthus cirrhifer)

File fish, Monacanthus cirrhifer, juveniles with initial mean body weight of 0.27g were fed purified diets with or without calcium (Ca) supplement for 10 weeks at a water temperature of 27.0±1.4°C. Growth was significantly low in fish fed diet without Ca supplement than fish fed diet with Ca supplement. Feed efficiency and condition of fish were also significantly decreased in absence of a dietary Ca supplementation, Minerals contents of bone were similar in both the treatment groups and did not appear as a suitable indicator of Ca requirement. It appeared that Ca supplement to the purified diet is necessary for file fish for their proper growth and feed utilization.

Effects of dietary vitamin E on the growth and breeding performance of Ompok pabda

A 90 day feeding trail was conducted to investigate the effect of vitamin E on the growth and breeding performance of Ompok pabda. A total of 84 healthy female brood fish (41.10±0.44g) were divided into 4 treatments i.e. treatment T1 T2, T3 and T4 having three replications each. The fish were fed twice a day with a standard feed (40% protein) having 4 doses of vitamin E viz. 0 (served as control), 50, 100 and 150mg vitamin E/kg feed. At the end of the feeding trial, the brood fish were induced to breed with PG extract to observe the effect of vitamin Eon feed. After rearing for 90 days with the experimental feeds, it was found that weight gain and specific growth rate of brood fish fed with 100mg vitamin E/kg feed (treatment T3) was the highest (14.78±0.38g and 2.99±0.11) while 150mg vitamin E/kg feed (treatment T4) fed fish gave the poorest result (2.97±0.89g and 1.21±0.32). There was no significant difference in terms of length gain of brood fish among the different treatments. The brood fish were induced to breed with equal dose of PG extract (18 and 12mg PG/kg body weight for female and male respectively) to observe the dietary effect of vitamin E on breeding performance. The highest ovulation, fertilization and hatching rate of eggs were found to be 81.48±6.41%, 84.04±3.53% and 68.59±5.03% respectively in the brood fish of treatment T1 while the poorest (33.33±00%, 52.35±5.02% and 45.70±7.24% respectively) were found in the brood fish under treatment T4. The results suggest that inclusion of 100mg vitamin E/kg feed is best for enhancing the breeding performance of O. pabda brood fish indicating that vitamin E content has a positive impact on reproduction of fish. The present results also imply that inclusion of higher level of vitamin E exerts an antagonistic effect in terms of growth and breeding performance of this species.

Dietary protein and energy interations: an approach to optimizing dietary protein to energy ratio in walking catfish, Clarias batrachus

An 8 weeks feeding trial was conducted in a static indoor rearing system to investigate protein to energy ratio (PIE ratio) in walking catfish Clarias batrachus. Six fishmeal based diets of two protein levels (25 and 35%), each with three lipid levels (5, 10 and 15%) resulted in P/E ratios ranging from 13.57 to 21.97 mg protein kJˉ¹ gross energy (GE) were fed to 50 fish in triplicate. Fish were fed 6% of their body weight three times per day adjusted fortnightly. Significantly higher (p<0.05) growth rates in terms of weight gain, % weight gain and specific growth rate (SGR) were evident in fish fed with higher protein diet. The highest growth rate was found by fish fed 35% protein, 17.06 kJˉ¹GE with a P/E ratio of 20.55 mg protein kJˉ¹GE. Significantly better (p<0.05) feed conversion ratio (FCR) was also evident in fish fed with higher protein diet and best FCR was found by fish fed 35% protein, 10% lipid, 17.06 kJˉ¹GE with a P/E ratio of 20.55 mg protein kJˉ¹GE. Significantly indifferent (p>0.05) values of protein utilisation were found in between the both (higher and lower) protein diets. Higher lipid deposition (p<0.05) in whole body was observed with increasing dietary lipid level at each protein diet and as higher (p<0.05) for the lower protein diets. The study reveals that C. batrachus performed best the diet containing 35%, 17.06 kJ gˉ¹ and 20.55 mg protein kJ gˉ¹ GE protein, gross energy and P/E ratio respectively.