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.

The food and feeding habit of Penaeus monodon Fabricius collected from Makato River, Aklan, Philippines

The food of Penaeus monodon collected from Makato R., from Sept 1977 to Jan 1978 is described with preliminary observations on its feeding habit and rate of foregut clearance. Feeding behaviour appears to be associated with the tidal phase. Foregut clearance rate is rapid, with 95% of food transported from the foregut 4 h after feeding. Frequency of occurrence and proportion of total food of various foregut contents are shown, as are dry weight, percentage mineral, organic and crude protein nitrogen from individual and pooled samples of gut contents, and foregut index in P. monodon collected during different phases of one tidal cycle.

The use of decapsulated brine shrimp eggs as food for shrimp larvae

Decapsulated cysts of Artemia salina were experimentally fed to the larvae of Penaeus monodon, Metapenaeus ensis, M. endeavouri and Macrobrachium rosenbergii and were found to be comparable to freshly hatched brine shrimp nauplii.

Preserved algae as food for Penaeus monodon larvae

Uni-algal cultures of C. calcitrans, S. costatum, T. chui and Isochrysis sp. obtained from the laboratory were harvested. The harvest, preserved by either freezing or sun-drying, was fed to the larvae of P. monodon . Among the test algal species, Chaetoceros and Tetraselmis were used in larval feeding trials with frozen food while Chaetoceros, Tetraselmis and Isochrysis species were utilized in feeding experiments with sun-dried algae. Their relative effects on larval survival and development were assessed. Results showed that, except the alum-flocculated cells, both frozen Chaetoceros and Tetraselmis can support survival at the zoea stage. Best survival of 68% was atained with dried Chaetoceros followed by Tetraselmis at 44%. Dried Isochrysis did not perform as well, a significantly low survival of only 25% was obtained.

Viability of frozen algae used as food for larval penaeids

Freezing with added chemicals as flocculants and protectants was assessed as a means of preserving stock cultures of 4 algal species used for larval penaeid food Chaetoceros calcitrans, Skeletonema costatum, Tetraselmis chuii and Isochrysis galbana . The maximum storage effectability of the preservation techniques for each species was also determined.

Culture experiments on the copepod, Tisbintra elongata MORI, and evaluation of that species as food organism for milkfish larvae

Five types of food were used to culture T. elongata: rice bran, cow dung, bread, cow dung, bread yeast, fermented fish solubles and Spirulina. Bread yeast was found to give the highest densities. An evaluation was also made of the effectivity of rice bran and fermented fish solubles for the outdoor mass culture of T. elongata . A comparative study on the growth and survival rate of milkfish fry (Chanos chanos) fed with T. elongata and Artemia showed there to be no significant differences between the diets.

Growth phases of cultured algae used as larval food

A study was undertaken to determine the onset and duration of the growth phases of cultured algae commonly used as larval food (Skeletonema costatum, Chaetoceros calcitrans, Tetraselmis chuii, Chlorella vulgaris, Isochrysis galbana) so as to predict the time of harvest at the desired stage to suit various needs and purposes.

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.

Feeding behavior and food preference of Penaeus monodon Fabricius with scrap tilapia

The time of day during which P. monodon feeds at different depth levels in earthen ponds, and its preference for three types of tilapia feeds (dry, fresh and fermented) were determined. It was observed that P. monodon concentrated at the bottom beds during the day and along the periphery of dikes during night-time, with a slight tendency to swim and feed towards the surface as darkness increased. P. monodon showed special preference for dried tilapia compared to fresh and fermented tilapia. P. monodon also showed adaptability to the platform method of feeding, especially during night-time.

Viability of frozen algae used as food for larval penaeids

Freezing with added chemicals as flocculants and protectants was assessed as a means of preserving stock cultures of 4 algal species used for larval penaeid food Chaetoceros calcitrans, Skeletonema costatum, Tetraselmis chuii and Isochrysis galbana. The maximum storage effectability of the preservation techniques for each species was also determined.