| Whitmania pigra Whitman belongs to Haemo pidae and dried W. pigra, Hirudo, is a famous medicinal material to treat cardiovascular and cerebrovascular diseases.Huge demands in the market of Hirudo and increasing environment pollution leads to the wild resources getting seriously reduced. Although the cultivation of W. pigra has got success, but its development was limited because of shortage of understand about digestive tract and reproductive organ. In this paper, the development of W. pigra digestive tract and reproductive organs was studied using histological, histochemical, and detect enzyme activity methods to do some primary work for its better knowledge. Results show that:(1) Mouthpart developmental histology of W. pigra at different month of age were studied by paraffin section, H.E staining combined alcian blue and periodic acid Schifts reaction procedure (AB-PAS). Results:Change ranges:oral width 0.6 mm (1-3 month); 1.2 mm (3-4 month); Oral diameter 0.3 mm (1-3 month); 1.2 mm (3-4 month), the oral size unchanged reached maximum during 4-6 months. Oral lip has thin protective film located in the front of the mouthpart. The W. pigra have three jaws in oral cavity, the big one is in dorsum; the other two separated on both side of abdomen respectively. Jaws and muscular pHarynx are interrelated closely. The jaws were composed by cuticle, epithelial layer, muscularis and jaw cavity from outside to inside. In the front of jaws have mastoid abdomen with function of secreting acidopHilic granule from 2 month age. Oral cavity was composed by mucosa, submucosa and muscularis inside and out. In oral cavity were rich of peristomial nerves. And pharynx was composed by mucosa, muscularis, adventitia inside to out. The folds height and width changed heighten and thickening. Mucosa epithelium from complex flat epithelium changed into columnar epithelium, muscularis gradually developed into thickened along with growing. Muscular thickness reached maximum at 4 months. Mucous cells of W. pigra were classified into Ⅰ-Ⅳ types based on different staining and two mainly morphological shapes (Tubular, Pear-shaped). Jaws, oral cavity, pharynx by AB-PAS staining showed little changes at different month of age. Mucous cells were few at 1 month age, and type Ⅱ cells were increased rapidly in 2-3 month age in oral lip. Oral cavity contains more mucous gland cells type Ⅰ. Under the muscularis were connective tissues which distributed a few of mucous cells type Ⅱ.(2) This research application tissue slice morphology observation and the light microscope measurement technique study the digestive system of W. pigra at different months. The results indicated that the digestive tract structure of W. pigra had highly differentiated when these were born, but the fuction is not perfect.1-2 months belongto the stage of development,2-3 months belong to the stage of approached perfection,4months belong to the stage of mature. Marked feature of the digestive system development is that the change of the type of epithelium cell of alimentary track (the chang from epithelial cells, short column cell to high column cell), the increasing of richness and altitude of mucous membrane fold and Increase in the number of goblet cell. Conclusion:Development characteristics of digestive tract in W. pigra coincided with the ingestive way of W. pigra and the function of digestive tract, which further proves that regardless of growth stage, morpHology and histology of digestive tract were closely related to its function.(3) Analysis and determination the activities of four digestive enzymes (Amylase, Lipase, Acid protease and Alkaline protease) of W. pigra at different months. The results of experiment were as follows:The activities of digestive enzymes presented a series of changes with the increase of month ages. The activities of Acid protease maintained at low level at 1-2 months, from 2 months the activities increased sharply to the peak at 2.5 months. Then cut down sharply to very low levels. The activities of Alkaline protease at low level at 1-2 months, from 2 month ages the activities reached the maximum point, then decreased sharply to low levels, at 3 months age the activies increased by a small margin, then decreased to low levels. From 1-3 month ages, the amylase activities increased progressively with the increasing of months. At 3 month ages reached the maximum point then followed by a decrease to low levels. The lipase activities decresed with the increase of age from 1-1.5 month ages, after 1.5 month ages the activites sharply to the maximum point at 3 month ages, then decreased sharply to low levels, at 3 months age the activies increased by a small margin, then decreased to low levels.(4) Objective to investigate the distribution of acid phosphatase (ACP), alkaline phospHatase (ALP), Lipase (Lipase) and nonspecific esterase (NSE) in the digestive tract of W. pigra at different months by the method of enzyme histochemical. Results ALP were distributed in the all the parts of the digestive tract except for rectum. ALP increased with the increase of month ages. Different digestive ALP showed significant differences at the same month ages (crop> intestine> esophagus> the sixth side blind sac). ALP was mainly located in the mucosal epithelia and striated border. ACP was distibution on all the parts of the digestive tract. ACP increased with the increase of month age. Different digestive ACP showed significant differences at the same month ages (intestine> crop> rectum> esophagus> the sixth Side blind sac). ACP was mainly located in the striated border, mucosal epithelia and Lamina propria. As the developmental stages, the NSE shows apparent upward trend at each part of the digestive (crop> intestine> the sixth side blind sac> esophagus> rectum). Observe though slice we found mucosal epithelia, striated border and Submucosa have oberviously specific reaction. The changed of lipase different from three other digestive enzymes. The lipase in different digestive at differetnt month ages showed great different:Esophagus substantially showed increased in first, then decreased. While Crop showed that decreased first, then increased. Intestine and rectum have obviously upward trend. Side blind sac change showed that at first decreased then incresased at last decreased again. At same developmental stages of digestive lipase activity expressed as:(crop> intestine> rectum> esophagus> the sixth side blind sac). Though slice we found the lipases located in mucosal epithelia. The conclusion is that the activities of the four digestive enzymes in W. pigra chang obviously, implying the digestive function will be completed progressively with its development. Crop and intestine is the major digestive organs of digestion and absorption in W. pigra. While side blind sac may play a role only in storage.The highest enzyme activity located in mucosal epithelia(5) The reproductive system and gonad development and germ cell occurrence of W. pigra have been studied by using tissue section techniques.Through tissue section observation, the results showed that W. pigra is androgynism, and the male reprodudcdtive system is mainly composed of spermary, seminal vesicle, prostate, deferent duct and the penis. The female reproductive consisted of the ovary, fallopian tubes, vaginal capsule and albumen gland.The development of cycle of the gonad could be divided into 5 stages: resting stage, proliferating stage, growing stage, maturing stage and spawning stage according to development of reproductive cells in spermary and ovary. The development of the sperm was divided into 5 stages including primary sper matocytes, secondary spermatocytes, spermatozoa, and sperm. The development of the ovum was divided into 4 stages including oogonia stage, no yolk cell stage, Vitellogenesis stage and film formation stage. The development of testes was earlier than the ovaries. |
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