Seasonal changes in serum leptin of the feral raccoon (Procyon lotor) determined by canine-leptin-specific ELISA

Several reports have been published on blood leptin concentrations in feral animals, including members of the Carnivora, using a commercially available multi-species radioimmunoassay (RIA) kit with anti-human leptin antibody. However, we observed weak immunoreactivity between recombinant canine leptin and anti-human leptin antibody, suggesting a limitation in the applicability of the RIA kit for leptin assays in Carnivora species. We tested the applicability of RIA and sandwich enzyme-linked immunosorbent assay (ELISA) with anti-canine leptin antibody to assay blood leptin in the dog (Canis familiaris) and the raccoon (Procyon lotor). When RIA was used for recombinant canine leptin and dog sera, values were much lower than those determined by ELISA at higher concentrations (>10 ng/ml), while rather higher at lower concentrations (<2 ng/ml). A similar discrepancy between the two methods was found for serum leptin concentrations in raccoons. Clear seasonal variations were observed by ELISA, but not by RIA, with high values in autumn (3.46+/-0.45 ng/ml) and low values in spring and summer (0.71+/-0.07 ng/ml). Serum leptin concentrations in raccoons correlated positively with their body weight (r=0.753) and body mass index (r=0.755), corroborating our previous findings of a strong positive correlation between serum leptin concentrations and body fat content in dogs. Thus, the canine leptin ELISA is useful for assays of dog and raccoon leptin, and blood leptin is a good marker of nutritional condition in the species of Carnivora assayed in this study.     

Phylogenetic relationships among deer in China derived from mitochondrial DNA cytochromeb sequences

The phylogenetic relationships of Cervidae and Moschidae were examined using partial sequence data of mitochondrial DNA (mtDNA) cytochromeb. Ten new sequences were obtained for six species of Cervidae and Moschidae, and aligned with those previously reported for other deer species. Our results demonstrated that the phylogenetic status of the taxa inferred from molecular data was congruent with taxonomy based on morphological studies. Cervidae formed a monophyletic group that consists of four subfamilies: Cervinae, Muntiacinae, Hydropotinae, and Odocoileinae. Moschidae occurred at the base of the Cervidae clade. On the basis of molecular clocks for genetic distance, the divergence time of mtDNA haplotypes within the subfamily Cervinae, among subfamilies in Cervidae, and between Moschidae and Cervidae was estimated to date 2–7 MYA, 6–10 MYA and 8–13 MYA, respectively.     

Determination of age in the Japanese monkey from growth layers in the dental cementum

The teeth of 14 Japanese monkeys (Macaca fuscata) were examined to establish an exact method of determining age by histological observation of dental cementum. The cementum showed annual growth layers, which were especially remarkable in the incisor root and in the molar cementum deposited at the junction of the roots. The layer of cementum formed in winter appears as a dark layer in stained sections and as a translucent layer in unstained ground sections. In the incisor the first dark and light layers are formed at the age of three years, whereas in the molar they do not appear at a definite age. The layers are thick and clear in the upper medial incisor. As a result, the age of a Japanese monkey can be determined by adding two to the number of dark layers and an outer light layer. It is interesting that the formation of the cementum of the first molar begins a few years after its eruption. The relation between this fact and the pressure of occlusion is discussed.     

Application of molecular techniques to distinguish Liriomyza trifolii from L. sativae (Diptera: Agromyzidae) on tomato cultivation in Japan

A molecular method is applied for differentiating the morphologically related leafminers Liriomyza trifolli and L. sativae on tomato cultivation. The method requires multiplex polymerase chain reaction (PCR) amplification of a region of mitochondrial cytochrome oxidase DNA using multiprimer sets. The method divides the mitochondrial fragment of L. trifolli into two fragments and L. sativae into three fragments. It is faster, less costly, and easier than random amplified polymorphic DNA-PCR, PCR-restriction fragment-length polymorphism, and DNA sequencing and more sensitive than the enzyme electrophoresis method, which are other ways to differentiate these two species. We applied the method to samples from populations of another place, sex, and stage and obtained the same results.     

Growth-related changes in histology and immunolocalization of steroid hormone receptors in gonads of the immature male green turtle (Chelonia mydas)

Studies on the population dynamics of sea turtles require histological evaluation of the ontogenetic development and the activity of the gonads for reproduction. To investigate the growth-related changes of gonads in the immature male green turtle (Chelonia mydas), the histological changes of testes and epididymides and the localization of the androgen receptor, estrogen receptor alpha, estrogen receptor beta, and progesterone receptor were examined. The testes were categorized histologically into six developmental stages, and a scarce relationship between straight carapace length and gonadal development was confirmed based on the histological analysis. Several kinds of steroid hormone receptors were examined to show distributions in both testes and epididymides, for which their immunoreactivities were enhanced according to the developmental stage of the testes. These results suggest that straight carapace length is not an adequate indicator of maturity determination, whereas histological and immunohistochemical evaluations are useful in identifying the growth stages of green turtles, owing to the higher sensitivity to steroid hormones that appear during growth.     

A case of cleft palate in a Kuril seal (Phoca vitulina stejnegeri), from Hokkaido, Japan.

A male pup Kuril seal (Phoca vitulina stejnegeri) from Hokkaido, Japan, was observed with a unilateral and total cleft of the primary palate. Complications included a supernumerary tooth, hypoplasia and asymmetry of the face, and deformation of the eyeball and external auditory meatus. An accompanying pneumonia may have resulted from water flowing into the respiratory system due to imperfect closure of the nostril by the cleft. No other abnormalities were found.     

中国新疆马鹿亚种头骨形态的地理变化

新疆马鹿（Cervus elaphus）有3个亚种（塔里木亚种C．e．yarkandensis、天山亚种C．e．songarieus和阿勒泰亚种C．e．sibirieus）。本文报道新疆马鹿3个亚种头骨形态学特征描述和14项形态学指标测定。被测定的指标主要有颅全长（GLC）、颜面长（FL）、鼻骨全长（GLN）、前头长（MFL）、眼窝长（OFW）、基底全长（CBL）、口盖最大幅（GPB）、前臼齿间距离（Pm—P）、眼间最小幅（ZB）、筋突起高（CH）、关筋突起高（LH）、牙齿间隙裂高（DH）、前下颚骨高（HMP2）、后下颚骨高（HMM,）,采用主成分分析并在这3个亚种之间进行了比较。结果表明：1）阿勒泰亚种的头骨长度和高度值比塔里木亚种大,宽度值比塔里木亚种小;2）天山亚种的头骨大小一般处在塔里木亚种和阿勒泰亚种的中间,但三者之间没有显著性差异;3）阿勒泰亚种和天山亚种的头骨形态测量数据比较接近,塔里木亚种的头骨形态比较特殊（塔里木亚种的头骨短而宽,明显区别于其他两亚种的特征）。在新疆导致马鹿3个亚种头骨形态差异的主要因素有栖息地环境和气候因素的差异。阿勒泰亚种和天山亚种栖息在大陆性气候寒冷区的针叶林中,冬天气候比较寒冷;塔里木亚种栖息在大陆性气候温热区的塔里木河流域胡杨林中,气候干旱炎热。因此,阿勒泰和天山亚种在适应寒冷的气候并适应生软食物的食性过程中头骨变成长细,而塔里木亚种长期适应干旱炎热并适应生硬食物的食性使头骨变的短而宽。     

Population genetic structure of the spotted seal Phoca largha along the Coast of Hokkaido,based on mitochondrial DNA sequences

Population genetic structure of the spotted seal, Phoca largha, along coastal regions of Hokkaido was investigated, using mitochondrial DNA (mtDNA) sequences. A 571-bp fragment of the mtDNA control region and adjacent threonine and proline transfer RNA genes was sequenced from 66 seals. We categorized all individuals into three groups considering sampling area and season: twenty-four seals from the Sea of Okhotsk in winter, 11 seals from the Sea of Okhotsk coast in fall, and 31 seals from the Sea of Japan coast in winter. From the 66 animals, 57 haplotypes were identified. Compared with the harbor seal sequence, all spotted seals examined shared two deletions in the control region, which distinguished between the two species. Forty-nine haplotypes were represented by a single individual, and haplotypes shared by more than two animals were generally restricted to same sampling-groups. Phylogenetic trees did not indicate clear geographic differences among the three groups. An Analysis of molecular variances (AMOVA) did not showed any significant population genetic structure in Hokkaido spotted seals (Phi(st )= -0.003). Our results showed a high level of diversity but no genetic structure, and did not deny the possibility that seals in the Okhotsk breeding concentration mainly stayed in the fall Okhotsk and also inhabited in the winter Sea of Japan.     

Ecological studies on a dragonfly,Nannophya pygmaea ramber (Odonata: Libellulidae) I. seasonal changes of adult population and its distribution in a habitat

Adult population of a dragonfly Nannophya pygmaeaRamber inhabited in a damp ground was investigated with mark-and-recapture method in 1975. The following results about the seasonal changes of the population size and distribution in the habitat were obtained. Adults emerged from late May to mid August. The number of the adults was most abundant in early June, but that of matured males in early July. From the recapture data, the estimate of daily survival rate was 0.82, and the length of immature stage in males was estimated as 5 days or so. Total number of post-teneral adults emerged in the habitat was estimated as about 9,000. The mean crowding-mean density regression method was applied for the analysis of the distribution pattern of the adults. Matured males showed a spaced-out distribution, while females and immatured males distributed themselves rather aggregatively. Such a distribution pattern of the matured male would be attributed to their territorial behaviour. The territorial behaviour was considered to force the matured males to extend the distribution area in July when they were most abundant. From the above-mentioned results and some observations, the meaning of the territoriality in this species was discussed.     

Molecular phylogeography of the red deer (Cervus elaphus) populations in Xinjiang of China: comparison with other Asian,European, and North American populations

To illustrate phylogeography of red deer (Cervus elaphus) populations of Xinjiang, we determined their mitochondrial DNA (mtDNA) control region sequences, and then investigated geographic variations and phylogenetic relationships between Xinjiang populations and other populations from Asia, Europe, and North America. The C. elaphus mtDNA control region shared different copy numbers of tandem repeats of 38 to 43-bp motifs which clearly distinguished the Western lineage from the Eastern lineage of this species in Eurasia. The western lineage comprised the Tarim populations from southern Xinjiang and the European populations, all of which had four copies of the motifs. By contrast, the Eastern lineage consisted of populations from northern Xinjiang (Tianshan and Altai Mountains), other Asian areas (Alashan, Gansu, Tibet, Mongolia, and northeastern China), and North America, all of which shared six copies of the motifs. MtDNA phylogenetic trees showed that there are two major clusters of haplotypes which referred to the Western and Eastern lineages, and that subgroupings of haplotypes in each cluster were congruent with their geographic distributions. The present study revealed that a boundary separating the Western lineage from the Eastern lineage occurs between Tarim Basin and Tianshan Mountains in Xinjiang. Meanwhile, North American populations were genetically closer to those of northern Xinjiang, northeastern China, and Mongolia, supporting that C. elaphus immigrated from northeastern Eurasia to North America through the glacier-induced land-bridge (Beringia) which had formed between the two continents after Late Pleistocene.