Life‐history strategies indicate live‐bearing in Nothosaurus (Sauropterygia)

In Sauropterygia, a diverse group of Mesozoic marine reptiles, fossil evidence of viviparity (live‐bearing) only exists for Pachypleurosauria and Plesiosauria, and was assumed to also be the case for nothosaurs. Previous studies have successfully applied an extant squamate model to sauropterygian life‐history traits. In extant squamates, oviparity and viviparity are associated with differences in life‐history trait combinations. We establish growth curves for Nothosaurus specimens based on their humeral histology. We then analyse life‐history traits derived from these curves and compare inferred traits to those of modern squamates and pachypleurosaurs to assess their reproduction mode. We show that birth to adult size ratios (i.e. birth size divided by the mother's size) provide good estimates of clutch sizes in extant squamates and in viviparous extinct marine reptiles, but these ratios cannot discriminate viviparous and oviparous squamates. Thus, large ratios do not indicate viviparity in fossil taxa to which the extant squamate model is applicable. Applying differences in birth size, age at maturation, and maximum longevity that are observed between extant viviparous and oviparous squamates to our Nothosaurus sample, we identified 7 out of 24 specimens as being potentially viviparous. Conversely, they suggested oviparity for many nothosaurs but also for many pachypleurosaur samples. Under the assumption that the entire clade Pachypleurosauria was viviparous, the majority of nothosaurs would also have been viviparous as they comprised trait combinations similar to those seen in pachypleurosaurs. Overall, this suggests that within nothosaurs and pachypleurosaurs both reproduction modes existed in different taxa.     

Gonad morphology and reproductive cycle of the mangrove oyster Crassostrea brasiliana (Lamarck, 1819) in the baía de Guaratuba,Paraná, Brazil

This study aimed to describe the gonadal histology and the reproductive cycle of Crassostrea brasiliana in the mangroves of Guaratuba Bay in southern Brazil. Adults were collected monthly from January 2010 to April 2011 from three sampling sites in intertidal oyster beds. The animals were evaluated using biometric and histological analyses of the gonads. The gonadal tissue samples were processed according to the standard histological procedures, and permanent slides were prepared using Harris' haematoxylin and eosin. The oysters were identified at the species level using a molecular protocol. Females (69%) predominated over males (26%), with 4% indeterminate and 1% hermaphroditic. Mature females were more prevalent in February, March and December 2010 and in March 2011. Mature males were more prevalent in February and April 2010 and in March 2011. The presence of hermaphroditic individuals was sporadic, and oysters in immature stages or sexual repose were observed in only a few collections between the months of May and October 2010. The reproduction of C. brasiliana in Guaratuba Bay occurs intermittently, but with greater intensity during the summer, with a larger number of females produced.     

Reanalysis of the phylogenetic status of Nipponosaurus sachalinensis (Ornithopoda: Dinosauria) from the Late Cretaceous of Southern Sakhalin

Nipponosaurus sachalinensis is the only definitive lambeosaurine hadrosaurid from Sakhalin Island of Russia. Previous studies suggested it was a member of Lambeosaurini (derived lambeosaurines). However, its phylogenetic status within Lambeosaurini remains controversial. In addition, some studies argued the juvenile ontogenetic stage of the holotype and regarded Nipponosaurus as an invalid taxon. In order to solve these problems, its definite growth stage is determined through histological studies. Absence of a line of arrested growth, presence of osteons with large vascular spaces, and presence of primary bone remnants even in the highly modified regions of the femur confirm that the holotype was a juvenile. More than a hundred of the 350 characters used to determine the phylogenetic position of Nipponosaurus are ontogenetically variable characters based on the different ontogenetic stages of Hypacrosaurus stebingeri. Our phylogenetic analysis reveals that Nipponosaurus is a basal lambeosaurine hadrosaurid, much further down in the tree than previously suggested, and shows a polytomy with Blasisaurus and Arenyisaurus. This study also indicates that Nipponosaurus is a valid taxon because it possesses unique characters within the Lambeosaurinae (presence of massive surangular anterodorsal process, presence of lateral shelf of the dentary, and a relatively short ulna), which are independent of ontogeny.     

Gonadal morphology in the self-fertilizing mangrove killifish, Kryptolebias marmoratus

We conducted anatomical and histological observations of the gonads in the self-fertilizing mangrove killifish, Kryptolebias marmoratus to investigate the self-fertilizing mechanism of this species. The gonad has a bilobed structure. The elongated gonadal lumen (GL) along the dorsal surface of the gonad connects to the common genital sinus. The elongate testicular region is closely attached to the GL. Among the ovulated eggs in the GL, those in the anterior part of the GL have micropyles but no perivitelline space (are not yet fertilized), whereas those in the posterior part of the GL are fertilized. In our histological analysis, we found free sperm in the posterior area of the GL. We conclude that ovulated eggs may be self-fertilized in the posterior GL.     

3D Culture Histology Cryosectioned Well Insert Technology Preserves the Structural Relationship between Cells and Biomaterials for Time‐Lapse Analysis of 3D Cultures

When performing histology of softer biomaterials, aspiration disrupts the cellular and molecular location information. This study aims to develop a cryosectionable well insert able to preserve the biomaterial and cell's original 3D conformation from the well to histology analysis. The well insert is composed of a paraffin‐coated gelatine pill. Within the coated capsule, the human epithelial cell line (NS‐SV‐AC) is cultured in Matrigel, GrowDex, Myogel, Myogel + GrowDex, or cell culture media for 14 days. At 0 and 14 days, the samples are frozen in liquid nitrogen and cryotome is used to create sections. The slides are stained by Sirius Red and immunohistochemistry using antibodies human collagens I–V and human Ki‐67. Sirius Red shows pink shades of biomaterials and the best cellular vertical distribution throughout the sagittal section of the well is achieved with Matrigel, GrowDex, and Myogel + GrowDex; in Myogel and media, the cells sink. For collagen protein expression, only Matrigel induces a notable difference while in the other materials, collagen staining is weak or difficult to distinguish from endogenous collagens. Ki‐67 expression is maintained over time. The 3D‐cryo well insert provides a new time‐lapse histology perspective of analysis for liquid or gel cultures that maintains cells and macromolecules in their unaltered in‐well configuration.     

海南龙血树的形态组织学研究

采用植物解剖学、显微切片技术等,分别对海南龙血树含有树脂的茎、不含树脂的茎、幼根和叶片等进行了系统的组织学研究。结果表明：老茎主要由栓化层、皮层、形成层和基本组织4部分组成,树脂主要分布在基本组织内维管束的导管和纤维中。叶片为等面叶,气孔主要分布在下表皮上、具有明显的孔下室,上下表皮内侧分布着大量的纤维束。幼根由根被细胞、皮层和维管柱组成。根、茎、叶的部分薄壁细胞中均含有晶束。海南龙血树营养器官的结构特征与干旱、高温和贫瘠的生态环境相适应。这些结果可为海南龙血树的开发和利用提供基本的解剖学证据。     

麝鼠香腺发育及腺细胞培养分离的初步研究

目前麝鼠香是麝香最好的天然替代物,麝鼠香来源于麝鼠生殖系统中的香腺,已发现其香腺中分泌细胞和支持细胞是麝鼠泌香的关键。本研究通过组织形态、HE染色、免疫组织化学染色及免疫荧光鉴定方法,初步描述了麝鼠香腺的发育过程。麝鼠香腺的形态结构显示其由小到大再至非泌香期萎缩;HE染色及组化结果显示,香腺在发育初期富含颗粒饱满的腺泡,雄激素分泌处于较低水平,分泌细胞数量较少,随着进一步发育,雄激素水平及分泌细胞数量逐渐升高,在两个月时达到最高,腺泡逐渐变大成熟并开始释放麝鼠香等分泌物;在泌香末期,腺泡逐渐被结缔组织取代。以上结果将有助于麝鼠香腺发育及分泌机制的研究,为提高麝香产量及实现体外泌香建立基础。另外,我们成功分离并鉴定了分泌细胞和支持细胞,以期为后续建立体外泌香体系奠定基础。     

Histology of gonads and cephalic brood pouch of the marine gastropod Planaxis sulcatus of Karachi Coast (Pakistan)

This histological study of 141 specimens of adult Planaxis sulcatus from Karachi showed thepresence of only female gametes in gonadal follicles,supporting the conclusion of earlier workers that thespecies may be parthenogenetic. The gonadal folliclesof several snails were occupied by an unidentifiedtrematode parasite.     

A brain atlas for the camouflaging dwarf cuttlefish,Sepia bandensis

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Multiple host feeding in field populations of Anopheles culicifacies and An. subpictus in Sri Lanka

A histological technique was used to detect the multiple blood-feeding of female mosquitoes, within the same gonotrophic cycle, in field populations of the malaria vectors Anopheles culicifacies and An. subpictus (Diptera: Culicidae) at a village in the low country of Sri Lanka, during 1994-96. Among 3306 An. culicifacies and 871 An. subpictus engorged females examined, respectively, 34.4% and 30.4% were multiple-fed. In these two species, double meals accounted for 92.7% and 89.5%, and triple meals for 7.3% and 10.5% of multiple meals, respectively. More females of An. culicifacies (53.7%) than An. subpictus (44.5%) multiple-fed on different nights. Multiple feeding rates in the two species were independent of sample size and field abundance. In An. culicifacies, but not An. subpictus, these rates were proportional to ambient temperature but not to humidity. Bloodmeal ELISA showed that 8.3% of 242 An. culicifacies were human-fed and 80% of these (i.e. 6.6%) were concurrently bovid-fed. Combining the histological and ELISA data, it was estimated that multiple feeding involving a human host had occurred in 7.2% of the blood-feeding An. culicifacies population at this study site. Only 1.2% of 406 An. subpictus were human-fed and all of these were concurrently bovid-fed. Multiple bloodfeeding within the same gonotrophic cycle was attributed to a local 'frequent feeding strategy' in these primarily zoophagic and endophilic malaria vectors. Unfortunately this strategy enhances their human-biting rate and hence their vectorial capacity.