一例人工圈养繁殖的马来穿山甲胎儿

一例人工圈养繁殖的马来穿山甲胎儿在华南师范大学穿山甲人工驯养与繁育研究基地出生。不幸的是2011年8月11日胎儿的母亲在胎儿快要临产时因病而亡,胎儿也因母兽病亡而夭折。雌性,全长19.8 cm,重120 g,已基本发育成熟,估计不久就会出生,已制作成标本与母兽标本一起保存在华南师范大学生命科学学院标本馆。胎儿的父亲仍饲养在华南师范大学穿山甲人工驯养与繁殖研究基地,健康状况良好。这是有报道的全球第一例人工圈养繁殖的马来穿山甲胎儿资料,十分珍贵。表明马来穿山甲在人工圈养条件下能够进行成功的繁殖交配,采取的人工驯养方法适合。     

马来穿山甲的年龄结构及寄生物

以广东湛江市林业公安局没收的51头马来穿山甲活体标本为材料,分析研究其年龄结构和寄生物。根据鳞甲、毛发、爪的磨损程度以及母幼行为关系和体重大小,将马来穿山甲划分成4个年龄组:幼年组、亚成年组、成年组和老年组,分别由7(13.7%)、21(41.2%)、19(37.3%)和4头(7.8%)个体组成,性比(♀∶♂)为1∶0.97。通过体表观察、粪检和解剖观察,发现被蜱感染过的穿山甲体况下降,且这些穿山甲仅有1种体外寄生物,为爪哇花蜱(AmblyommajavaneseSupino,1897),感染率为68.63%(35/51),未见幼体被感染。同时还讨论了该物种野放的生态安全问题。     

The regional distribution and relative frequency of gastrointestinal endocrine cells in SHK-1 hairless mice: an immunohistochemical study

The regional distributions and relative frequencies of some gastrointestinal endocrine cells in the eight portions (fundus, pylorus, duodenum, jejunum, ileum, caecum, colon and rectum) of the gastrointestinal tract of SKH-1 hairless mice were investigated using immunohistochemical methods and seven types of specific antisera against somatostatin, serotonin, glucagon, cholecystokinin (CCK)-8, secretin, pancreatic polypeptide (PP) and gastrin. In this study, somatostatin-, serotonin-, glucagon-, CCK-8-, secretin- and gastrin-immunoreactive (IR) cells were identified. Most of these IR cells in the intestinal portion were generally spherical or spindle-shaped (open-type cell) while cells that were round in shape (close-type cell) were occasionally found in the stomach regions. Their relative frequencies were varied according to each portion of gastrointestinal tract. Somatostatin-IR cells were found throughout the gastrointestinal tract except for the large intestine. Serotonin-IR cells were detected throughout the whole gastrointestinal tract and were the most predominant endocrine cell types in this species of mouse. Glucagon-IR cells were restricted to the fundus, occurring rarely. CCK-8-IR cells were observed in the pylorus, duodenum and jejunum with frequencies that were numerous, moderate and few, respectively. Peculiarly, secretin-IR cells were demonstrated in the whole intestinal tract with either few or rare frequencies. Gastrin-IR cells were restricted to the pylorus and were numerous. However, no PP-IR cells were found in this study. In conclusion, some peculiar distributional patterns of gastrointestinal endocrine cells were found in SKH-1 hairless mouse.     

The regional distribution and relative frequency of gastrointestinal endocrine cells in the nude mice, Balb/c-nu/nu: an immunohistochemical study

The distributions and frequencies of some endocrine cells in the eight portions of the gastrointestinal (GI) tract - fundus, pylorus, duodenum, jejunum, ileum, cecum, colon and rectum of the nude mouse, Balb/c-nu/nu were studied with immunohistochemical method using six types of anti-sera against serotonin, gastrin, cholecystokinin (CCK)-8, somatostatin, glucagon and human pancreatic polypeptide (hPP). All of six types of immunoreactive (IR) cells were identified. Most of IR cells in the intestinal portion were generally spherical or spindle in shape (open type cell) while cells showing round in shape (close type cell) were found in the intestinal gland and stomach regions occasionally. Their relative frequencies were varied according to each portion of GI tract. Serotonin-IR cells were detected throughout the whole GI tract and they showed the highest frequency in the pylorus. Gastrin-IR cells were restricted to the pylorus and duodenum with numerous and a few frequencies, respectively. CCK-8-IR cells were also restricted to the pylorus, duodenum and jejunum with numerous, a few and rare frequencies, respectively. Somatostatin-IR cells were demonstrated throughout the whole GI tract except for the colon and rectum, and they showed the highest frequency in the fundus. In addition, glucagon- and hPP-IR cells were restricted to the fundus and rectum, respectively with a few frequencies. In conclusion, the general distribution patterns and relative frequency of GI endocrine cells of the nude mouse, Balb/c-nu/nu was similar to that of other strains of mice. However, some strain and/or species-dependent unique distributions and frequencies of endocrine cells were also observed especially for somatostatin- and hPP-IR cells.     

Immunohistochemical study on the distribution of endocrine cells in the gastrointestinal tract of the babirusa, Babyrousa babyrussa (Suidae)

The distribution and relative frequency of endocrine cells in the gastrointestinal tract of the babirusa were studied immunohistochemically using the avidin-biotin-peroxidase complex method. Thirteen types of gut endocrine cells were detected; they were immunoreactive for chromogranin, serotonin, somatostatin, gastrin, bovine pancreatic polypeptide (BPP), glucagon, secretin, cholecystokinin (CCK), methionine-enkephalin-Arg6-Gly7-Leu8 (MENK8), motilin, gastric inhibitory polypeptide (GIP) and peptide tyrosine tyrosine (PYY). Cells that were immunoreactive for chromogranin, serotonin, somatostatin and glucagon were found in all portions of the gastrointestinal tract. MENK8-immunoreactive cells were observed in the stomach and small intestine. Gastrin-immunoreactive cells were detected in the pyloric region and duodenum. PYY-immunoreactive cells were found in the small and large intestine. Cells immunoreactive for motilin, CCK, GIP, and secretin were observed in the proximal small intestine and those immunoreactive for neurotensin were found only in the ileum. Although the distribution pattern of endocrine cells in the gastrointestinal tract of babirusa was similar to those reported for pig, restricted distribution of several endocrine cells, gastrin, BPP, MENK8, motilin, CCK, GIP, secretin and neurotensin and wider distribution of glucagon and PYY were observed in the babirusa. The unexpected presence of MENK8 in all glandular regions of the stomach and PYY in the small intestine was also noted. The distribution of gut endocrine cells might be related to the regulatory characteristics of the babirusa digestive tract.     

Immunohistochemical study of endocrine cells in the gastrointestinal tract of the barking deer, Muntiacus muntjak

The morphology, distribution and relative frequencies of endocrine cells in the gastrointestinal tract of the barking deer (Muntiacus muntjak) were investigated. The immunohistochemical method employed used seven types of antisera against chromogranin, serotonin, gastrin, cholecystokinin, somatostatin, glucagon and insulin. Chromogranin and serotonin immunoreactive (IR) cells were found throughout the gastrointestinal tract. Conversely, gastrin-IR cells were only found in the oxyntic gland, pyloric gland and duodenum, while cholecystokinin-IR and somatostatin-IR cells were detected in the oxyntic gland, pyloric gland and small intestines. Somatostatin-IR cells were also seen in the caecum. Glucagon-IR cells were found in all parts of the gastrointestinal tract apart from the colon and rectum. No insulin-IR cells were found in the gastrointestinal tract of this species. The cells in the small intestine were generally spindle shaped with long cytoplasmic processes ending in the lumen (open type), while in the stomach and large intestine, they were occasionally round or spherical in shape (closed type). An uncommon distribution pattern of endocrine cells in the gastrointestinal tract of the barking deer was noted for cholecystokinin- and glucagon-IR cells.     

Immunohistochemical study on the endocrine cells in gut of the stomachless teleost, Zacco platypus (Cyprinidae)

The regional distribution and relative frequency of neurohormonal peptides-producing cells were demonstrated in the gut of the stomachless teleost, Zacco platypus (Temminck et Schegel), using 10 types of specific antisera raised against mammalian regulatory peptides. The gut of Z. platypus was divided into five portions from proximal to distal (segment I-V). Most of immunoreactive cells in the epithelial lining portion, between epithelial cells, were generally spherical or spindle shape having long cytoplasmic process that was reached via the lumen while cells round in shape were found in the basal portions of epithelial lining occasionally. Serotonin-, somatostatin-, glucagon-, cholecystokinin (CCK)-8 and pancreatic polypeptide (PP)-immunoreactive cells were observed in this study. However, no chromogranin A-, secretin-, vasoactive intestinal peptide (VIP)-, substance P- and bombesin-immunoreactive cells were found. Serotonin-immunoreactive cells were demonstrated throughout the entire gut tract and occurred more frequently than other cells. Somatostatin-immunoreactive cells were restricted to proximal segments of the gut (segment I-III) with rare frequencies, and glucagon-immunoreactive cells were demonstrated in the proximal segments of the gut (segment I, II) with moderate to few occurrences. CCK-8-immunoreactive cells were found throughout the whole intestinal tract except for most proximal segment (segment I) with moderate to few frequencies and PP-immunoreactive cells were demonstrated in the proximal to middle segments, segment I-III, with a few, rare and rare frequencies, respectively.     

Immunohistochemical study of gastroenteropancreatic endocrine cells of the herbivorous Japanese field vole, Microtus montebelli

The gastroenteropancreatic (GEP) endocrine cells of the Japanese field vole were studied immunohistochemically. Somatostatin-, 5-hydroxytryptamine-, glicentin-, glucagon-, bovine pancreatic polypeptide-, gastrin-, gastric inhibitory polypeptide-, cholecystokinin-, substance P-, secretin-, neurotensin- and insulin-immunoreactive cells were revealed. The characteristic findings of the regional distribution and relative frequency of these immunoreactive cells in the GEP system of the vole were as follows. Somatostatin-immunoreactive cells were more numerous in the oxyntic glands than in the pyloric glands. Some somatostatin-immunoreactive cells were found in small clusters in the oxyntic glands. Gastrin-immunoreactive cells were detected not only in the pyloric glands and small intestine but also in the caecum and spiral colon. Gastric inhibitory polypeptide-immunoreactive cells were also detected in the pyloric glands and no motilin-immunoreactive cell was found in the gastroenteropancreatic system.     

马来穿山甲源嗜水气单胞菌亚种的分离鉴定及生物学特性分析

为鉴定马来穿山甲(Manis javanica)腿部致病菌并研究其致病性、耐药性和耐药基因,从救护的1只马来穿山甲腿部溃烂处采集样品,无菌分离培养致病菌,进行形态学观察、革兰氏染色及16S rRNA和gyrB基因测序分析,并开展细菌药敏试验和小鼠致病性试验,随后通过PCR扩增6种毒力基因(rtxA、earA、hlyA、alt、act、epa)及19种耐药基因(5种β-内酰胺酶相关基因、6种氨基糖苷类修饰酶基因、6种氯霉素耐药基因、2种磺胺耐药基因)。结果显示:分离的1株优势菌为革兰氏阴性短杆菌,经16S rRNA和gyrB基因测序以及序列进化树分析鉴定为嗜水气单胞菌亚种(Aeromonas dhakensis),其gyrB基因序列与马来西亚分离株高度同源,推测此致病菌可能由非法走私的穿山甲从马来西亚携带至我国。药敏试验显示,其对头孢曲松、头孢克肟、庆大霉素和氧氟沙星等药物敏感,对青霉素、阿莫西林、头孢氨苄、头孢克洛、氯霉素、磺胺甲恶唑、甲氧苄啶和林可霉素耐药。4种耐药基因tem、aac(3)-II、flor、sul2为阳性,与抗生素耐药表型相符。4种毒力基因earA、hlyA、alt、act为阳性,小鼠半数致死量为3.75×10^(6 )CFU/mL。根据药敏结果使用敏感抗菌药物治疗穿山甲,其腿部患处有好转。因此,引起穿山甲腿部溃烂的病原嗜水气单胞菌亚种(Aeromonas dhakensis)可能为国外传入菌株,具有较强的致病性和多重耐药性。本研究为嗜水气单胞菌的流行病学调查及其感染的防治提供了依据。     

Immunohistochemical studies on the development of endocrine cells in the thyroid and parathyroid glands in the golden hamster

The development of endocrine cells in the thyroid and parathyroid glands in the golden hamster was studied immunohistochemically in relation to the formation of these glands. The thyroid was formed on day 9 of gestation by the ventral outpocketing of the foregut between the first and second branchial pouches. The thyroid epithelial cells were faintly thyroglobulin-immunoreactive on day 10.5 of gestation. This immunoreaction became intense thereafter, but was almost confined to the cytoplasm of epithelial cells until birth. It appeared in the follicular lumen in newborn animals. The ultimobranchial body was derived from the fifth pouch and fused with the thyroid on day 12 of gestation. Calcitonin-immunoreactive cells first appeared on day 14 of gestation in the dorsomedial part of the thyroid derived from the ultimobranchial body and increased in number and intensity thereafter. Somatostatin-immunoreactive cells also appeared in the dorsomedial part of the thyroid derived from the ultimobranchial body on day 13 of gestation, and increased in number in newborn animals, but decreased thereafter. The parathyroid was derived from the third pouch, situated on day 13 of gestation on the dorsolateral side of the thyroid, and surrounded by a common capsule with the thyroid. Parathormone-immunoreactive cells first appeared on day 15 of gestation in the parathyroid and increased in number and intensity after birth.     

Immunohistochemical and ultrastructural study of endocrine cells from the pyloric region of the European mole (Talpa europaea)]

The distribution and the morphology of some endocrine cells (gastrin, somatostatin and 5-HT immunoreactive) in the pyloric region were studied in the Talpa europaea, an insectivore representing one of the most primitive living Eutherians. The immunohistochemical studies enabled us to identify and calculate the percentage of each cell type: the most numerous endocrine cells were gastrin immunoreactive; fairly numerous appeared somatostatin immunopositive; less numerous were 5-HT immunoreactive cells. While the ultrastructural observations let us describe four endocrine cell types: G cells producing gastrin, D cells containing somatostatin, EC cells of the gastric type producing 5-HT and D1 cells whose content is still unknown.     

Immunocytochemical distribution of endocrine cells in the gastrointestinal tract of the horse

Endocrine cells immunoreactive for somatostatin, gastrin, glicentin, glucagon, secretin, cholecystokinin, motilin and neurotensin were identified immunocytochemically in the gastrointestinal mucosa of the horse. Somatostatin-, glicentin- and glucagon-immunoreactive cells were very numerous in the cardiac and fundic regions of the stomach, whereas most gastrin-immunoreactive cells were confined to the pyloric region. Somatostatin-immunoreactive cells also were detected in all portions of the small intestine while gastrin-immunoreactive cells were confined exclusively to the upper portion and glicentin-immunoreactive cells were limited to the lower portions of the small intestine. Secretin-, cholecystokinin- and motilin-immunoreactive cells were observed only in the duodenum, while neurotensin-immunoreactive cells were confined primarily to the ileum. In the large intestine, somatostatin- and glicentin-immunoreactive cells were detected in the colon and rectum. The preferential location of endocrine cells provides additional information for future studies on the physiological roles of gastrointestinal peptides in the gastrointestinal tract of the horse.     

Immunohistochemical survey of the gut endocrine cells in the common tree shrew (Tupaia belangeri).

Regional distribution and relative frequency of endocrine cells in the gastrointestinal tract of the common tree shrew (Tupaia belangeri) were studied immunohistochemically. Ten types of immunoreactive endocrine cells were localized in the gastric mucosa, i.e., chromogranin-, serotonin-, gastrin-, somatostatin-, bovine pancreatic polypeptide (BPP)-, enteroglucagon-, pancreatic glucagon-, peptide tyrosine tyrosine (PYY)-, motilin-, and substance P (SP)-immunoreactive (IR) cells. In the intestine, 13 types of immunoreactive cells were observed, i.e., chromogranin-, serotonin-, somatostatin-, gastrin-, BPP-, enteroglucagon-, PYY-, secretin-, cholecystokinin (CCK)-, gastric inhibitory peptide (GIP)-, motilin-, neurotensin-, and SP-IR cells. The regional distribution and relative frequency of the cell types varied along the gastrointestinal tract. Basically, the types, distribution, and relative frequency of the gut endocrine cells were similar to those reported in other mammalian species. However, some characteristic findings were noted in the present study: (1) the considerably large number of gastrin-IR cells in the pyloric region; (2) numerous serotonin-IR cells in the stomach; (3) appreciable number of BPP-IR cells in the transitional region of the stomach; and (4) wide distribution of PYY- and motilin-IR cells in the gut.     

Comparative distribution of immunoglobulin-containing cells in stomach, intestine and associated lymph nodes of cattle

Immunoglobulin (Ig)-containing cells were investigated immunohistochemically in the stomach, intestine and associated lymph nodes of clinically normal calves, cows and fetuses to examine mucosal immune responses in the forestomach to rumen microbial flora. IgG-containing cells were observed in the mucosal propria of the forestomach of a 32 day-old, a 37 day-old, two 90 day-old calves, and of all 5 cases of cows. However, no IgA- and IgM-containing cells were observed. Further, no Ig-containing cells were detected in the associated lymph nodes of all calves and cows. Various numbers of Ig-containing cells, predominantly IgG, were observed in the mucosal propria of the abomasum, small intestine and cecum, and in the mesenteric lymph nodes of aged calves and all cows. No Ig-containing cells were observed in any tissues of fetuses. The results suggest that the development of mucosal immune responses in the forestomach may be incomplete as compared with that of the intestine.     

Quantitative study of pulmonary endocrine cells in fetal, postnatal and adult sheep

Using light microscopic immunohistochemistry, neuron-specific enolase (NSE)-positive endocrine cells were quantitatively analyzed in the sheep lung during different stages of development from the canalicular stages to adulthood. In all stages, NSE-positive endocrine cells were usually located in the bronchi and bronchioles as solitary cells, although a few NSE-positive cell clusters, the so-called neuroepithelial bodies, were found in some places. The number of NSE-positive endocrine cells decreased with advanced stages of gestation. In the late alveolar stage, the number of NSE-positive endocrine cells reached its bottom during the fetal period. There was a gradual upturn after birth. The overall pattern of growth and differentiation of the endocrine cells is most likely species-related and depends on the state of airway development; the number of the endocrine cells of almost all animals, excluding the sheep, in relation to the size of the lung reaches a peak in the late fetal and early neonatal periods and decreases shortly thereafter. NSE-positive endocrine cells were also predominantly located in the large airways during the early stage of development (canalicular stage), and were found more frequently in the small peripheral airways towards the term. These results show the number of NSE-positive endocrine cells in the sheep to be different from that seen in other species.     

Presence and distribution of neuroendocrine cells in the gastroenteropancreatic endocrine system of fallow deer foetuses

The gastroenteropancreatic endocrine system was studied in 11- and 17-week-old fallow deer foetuses using an immunocytochemical technique. In the gastrointestinal tract, gastrin-, serotonin-, somatostatin- and cholecystokinin-containing cells were found: their frequency and distribution were also determined. Anti-glucagon and anti-insulin antibodies did not stain any cells along the gut. In the pancreas, somatostatin-, pancreatic polypeptide-, insulin- and glucagon-immunoreactive cells were detected. The different distribution and number of neuroendocrine cells, in the two investigated stages of foetal life, are discussed. Data obtained in this study were compared with those published in a previous study on the gastrointestinal system of the adult fallow deer.