Comparative studies on lipid analysis and ultrastructure in porcine and southern minke whale (Balaenoptera bonaerensis) oocytes

The present study was conducted to clarify the difference in the color of the cytoplasm in immature follicular oocytes from prepubertal and adult minke whales. The four lipid contents (triglyceride, total cholesterol, phospholipids and non-esterified fatty acids) in vitrified immature oocytes from prepubertal and adult minke whales, and also in fresh and vitrified immature porcine oocytes, were measured. The lipid contents in vitrified-warmed minke whale oocytes were similarly high compared with those in vitrified-warmed porcine oocytes. In particular, the total cholesterol and phospholipid contents in the vitrified immature oocytes from prepubertal and adult minke whales were significantly (P<0.05) higher than those from prepubertal pigs. Furthermore, the distribution of lipid droplets in fresh and vitrified immature oocytes was observed in transmission electron microscopy. Lipid droplets in the prepubertal minke whale oocytes were distributed throughout the cytoplasm. In contrast, adult minke whales had larger lipid droplets which were distributed mainly in the central portion of the cytoplasm. The lipid droplets of immature oocytes from prepubertal pigs were larger than those in minke whale oocytes. These results indicated that the difference in the distribution of the cytoplasmic lipid droplets may result in the difference in the color tone of both prepubertal and adult whale oocyte cytoplasm.     

Microanatomy of the terminal air spaces of Baird's beaked whale (Berardius bairdii) lungs

The terminal airways and microvasculature of five adult Baird's beaked whales (Berardius bairdii) lungs have been examined by means of light and scanning electron microscopy of corrosion casts. The respiratory system of the Baird's beaked whale has various anatomical features which allow them to attain great depths and remain submerged for long periods. The whale lung has components including hyaline cartilage and smooth muscle throughout, reaching as far as the peripheral bronchi, sphincters surrounding the terminal bronchioles, the thick alveolar septa with a connective tissue core and a bi-layer capillary bed, and a distinctive venous plexus of the pulmonary veins. The well-developed venous plexuses of the pulmonary vein are found in the interlobular connective tissue, and around the airways and pulmonary arteries with close apposition. The hyaline cartilage throughout the airways may increase the effective dead air space that accommodates most of the air forced from the collapsed alveoli during a dive. The sphincter might serve as a cock for regulating buoyancy and for trapping air in the alveoli to prevent their complete collapse and a sucking in of alveolar tissue as the dive becomes deeper. The venous plexuses might be for pooling the large volume of blood in the lung to conserve oxygen for deep and prolonged diving.     

Phylogenetic analysis of a new Cetacean morbillivirus from a short-finned pilot whale stranded in the Canary Islands

Cetacean morbillivirus (CeMV) is considered the most pathogenic virus in cetaceans. Three strains have been already described: the dolphin morbillivirus (DMV), the porpoise morbillivirus (PMV) and the tentatively named pilot whale morbillivirus (PWMV).This study describes the molecular characterization of a strain of CeMV detected in the brain of a short-finned pilot whale that had stranded in the Eastern Atlantic Ocean around the Canary Islands and that showed lesions compatible with morbilliviral disease. Sequences for the nucleoprotein, phosphoprotein, fusion protein and haemagglutinin genes were obtained. The phylogenetic study showed high homology (97%) with the PWMV strain previously detected from a long-finned pilot whale stranded in the Western Atlantic Ocean. These results support the existing classification of CeMV into three principal genetic clusters.     

The pattern of ovarian development in the prepubertal antarctic minke whale (Balaenoptera bonaerensis)

This study describes the morphological and morphometrical changes associated with prepubertal ovarian development in the Antarctic minke whale (Balaenoptera bonaerensis). Ovaries were harvested from 94 immature minke whales caught in the Antarctic Ocean during the summer feeding season (December-March). Notable differences in ovarian size and morphology were found among animals. Up to 10 folds difference in ovarian weight was found among prepubertal whales of similar body size. During the prepubertal period, ovaries grew slowly and approximately doubled their weight. The morphologies of right and left ovaries were almost identical while the growth of the ovary appears to occur preferentially on the right side. The most striking morphological feature was numerous small antral follicles less than 5 mm in diameter found in ovaries of younger immature whales. The occurrence of these ovaries was highest in whales less than 6 m long and gradually decreased as body length increased. In larger whales, the occurrence of ovaries with a smaller number of follicles up to 10 mm and thick tunica albuginea increased. Thus, the ovary of the Antarctic minke whale experiences bursts of small follicular development during the early prepubertal period before becoming a more developed ovary with fewer but larger follicles, and thick tunica albuginea.     

Microvasculature in the terminal air spaces of the lungs of the Baird's beaked whale (Berardius bairdii)

Lungs were obtained from five adult Baird's beaked whales (Berardius bairdii) and examined by means of light microscopy and scanning electron microscopy of corrosion casts. The alveolar septa of these whales are thick with a connective tissue core and a bi-layer capillary bed. A double capillary network is regularly found in the alveolar duct and alveolar septa. Occasionally, septa adjacent to alveoli and alveoli themselves show only a single capillary layer. The distance between the two capillary layers has a tendency to decrease toward the end of airspaces, suggesting an end result of capillary fusion. Vascular replicas of venous vessels have annular furrows at regular intervals of 50 to 100 microm, which are caused by focal aggregations of collagen fibers circularly oriented and located immediately underneath the endothelium. The first valves appear in the collecting venules gathering alveolar capillaries. These valves are quite characteristic of flap-, funnel-and/or chimney like structures.     

Deep granulomatous dermatitis of the fin caused by Fusarium solani in a false killer whale (Pseudorca crassidens)

A 10-year-old female false killer whale (Pseudorca crassidens) developed skin lesions in the left breast fin. Histopathologically, the lesions consisted of multiple granulomas spread diffusely into the deep dermis and bone; characteristically, each granuloma had septate, branching fungal hyphae and chlamydospores surrounded by eosinophilic Splendore-Hoeppli materials. Macrophages, epithelioid cells and multinucleated giant cells in the granulomas reacted mainly to anti-SRA-E5 antibody against human macrophage scavenger receptor type I. Fusarium solani was isolated and its gene was detected from the skin samples. Mycotic skin lesions by Fusarium spp. reported so far in marine mammals were regarded as superficial dermatitis; therefore, the present case is very uncommon in that the lesions spread deeper into the skin.     

Attempt at in vitro maturation of minke whale (Balaenoptera Bonaerensis) oocytes using a portable CO2 incubator

The present study was conducted to investigate whether a portable CO2 incubator was effective for in vitro maturation (IVM) of bovine, porcine and minke whale oocytes, and the effect of maturation media supplemented with different hormones; porcine follicle stimulating hormone (pFSH), estradiol-17beta (E2), or pregnant mare's serum gonadotropin (PMSG): human chorionic gonadotropin (hCG) for minke whale immature oocytes was also examined. In vitro maturation rates of bovine and porcine oocytes cultured in the portable CO2 incubator were not significantly different from the standard CO2 incubator. In minke whale IVM culture using the portable incubator, the maximum expansion of cumulus mass was observed by pFSH/E2 and PMSG/hCG at the end of IVM culture. Moreover, the IVM culture period was shortened to 28-30 h from 96-120 h previously reported. The proportion of matured oocytes cultured in the medium supplemented with pFSH/E2 (26.7%) was significantly higher (P<0.05) than that with PMSG/hCG (6.9%). The present study indicates that a portable CO2 incubator is a useful device for minke whale IVM culture on a research base ship, and the addition of pFSH/E2 into an IVM medium enhanced cumulus expansion and the proportion of minke whale matured oocytes.     

Effects of inflammatory periodontal disease ('broken mouth') on the ultrastructure of collagen fibrils in the sheep incisor periodontium

The ultrastructure of the matrix of the sheep central incisor periodontium showing clinical signs of severe periodontitis was analysed quantitatively. The distribution of collagen fibril diameters in the lower dental pad changed from a bimodal distribution seen in healthy periodontia to a unimodal distribution. Collagen fibrils with an abnormal morphology were seen in the connective tissue adjacent to the crest of the alveolar bone. These results suggest that the deepening periodontal pocket resulting from inflammation removes the major area of support for the tooth and abnormal loads are applied to fibres deeper within the tissue.     

莱芜黑山羊松果体超微结构及5-羟色胺分布的季节性变化

应用5-羟色胺(5-HT)免疫组织化学方法和电镜技术,对莱芜黑山羊4个季节的松果体进行了研究.结果显示:(1)松果体内有大量5-HT阳性的松果体细胞及纤维;(2)松果体5-HT免疫阳性细胞面积以冬季最大,且与其他3个季节差异极显著(P<0.01);夏季最小;春秋季之间无明显差异(P>0.05).4个季节5-HT阳性细胞的光密度差异不大(P>0.05).松果体中5-HT阳性面积占总面积的比值.春、秋季相差不大(P>0.05),夏、冬季差异极显著(P<0.01);(3)松果体细胞有明细胞和暗细胞之分,2种细胞超微结构明显不同.明细胞胞核较大,常染色质多,粗面内质网(RER)、游离核糖体丰富,高尔基复合体的扁平囊泡层数较多,线粒体嵴发达,糖原较少;暗细胞的异染色质多,RER和游离核糖体较少,高尔基复合体的扁平囊泡层教少,线粒体嵴缺乏,糖原较丰富.结果提示,明细胞功能较暗细胞活跃;(4)不同季节松果体内明、暗细胞的数量不同,冬季为明细胞,夏季为暗细胞,春、秋2季2种细胞皆有,表明季节是影响松果体结构和功能活动的重要因素.     

Protective effects of WEB 2086 (PAF antagonist) and ketoprofen (NSAID) on PAF-induced changes in the morphological ultrastructure of blood platelets in calves

The ultrastructure of bovine platelets was examined by transmission electron microscopy without any pretreatment (control), and after WEB 2086 (a triazolodiazepine) or ketoprofen (NSAID) pretreatment, followed by PAF infusion. The blood platelet count was also investigated. The group of calves that received WEB 2086 pretreatment before platelet-activating factor (PAF) infusion did not show a decreased number of platelets. However, in the other group, with ketoprofen pretreatment before PAF infusion, there was a rapid decrease from 1 to 3 min, while from 5 min the number of platelets recovered to the normal value. Electron microscopy revealed that pretreatment with WEB 2086 followed by PAF infusion did not alter the morphological ultrastructure of bovine platelets, except that the microtubules were briefly modified from 1 until 3 min after PAF challenge. After ketoprofen pretreatment, bovine platelets kept their regular shape, the number of dense bodies was not significantly altered, the number of mitochondria was maintained from 5 min after PAF infusion, giant platelets were not observed and the Golgi apparatus was rarely visible. Thus pretreatment with WEB 2086 and ketoprofen before PAF infusion had a protective activity on the ultrastructure of bovine platelets and, in cattle, pretreatment with WEB 2086 and ketoprofen before PAF challenge prevented the thrombocytopenia induced by PAF.     

Mycobacterium marinum dermatitis and panniculitis with chronic pleuritis in a captive white whale (Delphinapterus leucas) with aortic rupture.

A 16-year-old female white whale, Delphinapterus leucas, died after nearly 18 months of chronic lymphopenia and pyogranulomatous dermatitis. Necropsy revealed rupture of the aorta with hemorrhage into the cranial mediastinum and between fascial planes of the ventral neck musculature. Multiple foci of ulcerative dermatitis and panniculitis were present across the thorax and abdomen and surrounded the genital folds. In addition, there was a chronic proliferative pleuritis with over 20 liters of histiocytic exudate in the thoracic cavity. Acid-fast bacteria consistent with Mycobacterium sp. were identified in sections of skin lesions and in cytospins of pleural exudate. Cultures of pleura and 1 skin lesion collected at necropsy yielded sparse growth of an acid-fast bacillus with colony characteristics and morphology consistent with Mycobacterium marinum. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis confirmed the presence of M. marinum DNA in samples of skin. This is the first documented occurrence of mycobacteriosis in a white whale and is a unique presentation of mycobacterial dermatitis and panniculitis with chronic pleuritis in a cetacean. The improved PCR-RFLP protocol utilized in this case unifies techniques from several protocols to differentiate between species of Nocardia and rapidly growing mycobacteria clinically relevant to aquatic animals.     

德国牧羊犬精子超微结构及超低温冷冻对精子超微结构的影响

为了研究超低温冷冻前后德国牧羊犬精子超微结构的变化,采用扫描电镜和透射电镜对冷冻前后德国牧羊犬精子超微结构变化进行研究,结果显示在扫描电镜下,鲜精整体呈“蝌蚪状”,头部呈扁卵圆形.在透射电镜下精子纵切面头部呈“楔形”,头部细胞膜由外向内分别由质膜、顶体外膜、项体内膜和核膜组成.精子尾部中段横切面由外向内可见线粒体鞘膜、9束外周致密纤维,9对轴丝和2根中央微管.精子头长约为5.40 μm,头宽3.26 μm,颈长1.25 μm,颈宽0.55 μm,尾部中段长11.34 μm,中段直径0.87 μm,尾部长55.70 μm,线粒体螺旋数为44旋.鲜精和冷冻前精子头部、颈部和尾部形态变化差异均不显著(P＞0.05).超低温冷冻后精子头部、颈部和尾部发生形态变化的精子数极显著高于鲜精和冻前精子数(P＜0.01).冻后顶体发生明显形态变化的精子占54.21％,高于发生颈部形态变化(10.61％)和尾部形态变化(28.83％)的精子数.结果表明鲜精和冻前精子形态变化不显著,而超低温冷冻后精子头部、颈部和尾部发生形态变化的精子数显著高于鲜精和冻前精子数.     

Classification of the parenchymal cells in Fasciola gigantica based on ultrastructure and their expression of fatty acid binding proteins (FABPs)

Parenchymal cells in adult Fasciola gigantica can be classified into three types based on their ultrastructural features and different quantities of fatty acid binding protein (FABP) being stored. Parenchymal cell type 1 (Pc1) has pale cytoplasm consisting largely of a loose network of fine fibers, and it contains few mitochondria but numerous glycogen particles. This cell type may be specialized in the storage and metabolism of glycogen and glucose. Parenchymal cell type 2 (Pc2) has similar cytoplasmic features as Pc1 but contains more numerous mitochondria, and high concentration of FABP as reflected by high density of immunostaining and immunogold labeling using specific monoclonal antibody (MoAb) to FABP as probe. Pc2 may, thus, specialize in the storage and metabolism of fatty acids and other lipids. Parenchymal cell type 3 (Pc3) has dense cytoplasm containing large amount of rough endoplasmic reticulum, Golgi complex and mitochondria, which is typical of a secretory cell. Furthermore, Pc3 has very little glycogen particles and is not stained by MoAb against FABP. It could, thus, be concerned with the synthesis of fibers, which form the scaffold of the parenchyma.