We adults should urgently restore natural habitats of wildlife to remain beautiful creatures to next generation children. Zoos and aquariums should make a great effort simultaneously for the longer term contribution to global biodiversity conservation by international cooperation with other conservation organizations. To ensure the survival of endangered species and to maintain their captive population number with genetic diversity, it is essential that great efforts should be made to establish assisted reproductive technique and cryopreserve the cells such as germplasm. Five zoos and Hokkaido University have been developing co-research on assisted reproduction in the Amur tiger and Hokkaido brown bear. As part of the time capsule project for genes and cells of endangered wildlife, tissue samples of threatened species were taken and transported to NIES for cell culture and cryopreservation. Primordial germ cells (PGCs) have been collected and cryopreserved from the eggs of captive birds. We also study on production of interspecific germline chimeras by use of the PGCs transplant operation for the purpose to proliferate endangered bird individuals in the future. Assisted reproduction and cell preservation is not directly contributed to wildlife conservation. It can be more important to announce these scientific attempts intelligibly to the general public, and to give them the opportunity to know the current status of wildlife as a tool of environmental education. In this review, study on assisted reproduction and cryopreservation of the cells from endangered wildlife is introduced, and zoo potentiality for wildlife and global health conservation is described.

View full abstract

One of many human-caused mortality factors that affect raptors is electrocution. Many raptor species perch on power poles or transmission lines, and electrocutions can occur. Electrocutions can affect endangered raptors that occur in Hokkaido, such as the Steller's sea eagle, White tailed eagle, Mountain hawk eagle, and Blakiston's fish owl in Hokkaido. It is important to increase our understanding of the reasons why some raptors are electrocuted and develop solutions to this problem. We estimate mortality due to mortality for a variety of endangered species, and examine the effectiveness of retrofitting electricity transmission infrastructure with devices that prevent electrocution. The need to reduce the threat of electrocution is urgent, and will not be achieved without the cooperation of the electricity company, wildlife biologists, government, and other stakeholders.

View full abstract

Mammary adenocarcinomas showing positive reactivity for estrogen receptor (ER), which were found in two female Western European hedgehogs (Erinaceus europaeus) over 5 years old, were successfully treated with surgical excision, but other malignant tumors occurred subsequently. In case 1 (cribriform carcinoma), the neoplastic tissue was characterized by formation of tubular spaces. In the majority of the tissue, there were large numbers of remaining myoepithelial cells, which were demonstrated distinctly by immunohistochemistry for alpha smooth muscle actin (SMA) and cytokeratin. A forearm tumor, which occurred 9 months after surgery, was diagnosed as myofibroblastic sarcoma, because the neoplastic spindle-shaped cells showed an SMA-positive and desmin-negative immunophenotype. In case 2 (comedo carcinoma), tumor cell nests showed central comedo necrosis, and lacked SMA-positive myoepithelial cells. Approximately 13 months after surgery, the animal died. Necropsy examination revealed that the animal had renal cell carcinoma, which was judged to be of proximal tubule origin on the basis of immunohistochemistry and electron microscopy.

View full abstract

Thirty-one ejaculates from 18 captive-living cheetahs (Acinonyx jubatus) maintained at three zoological institutions in Japan were collected by rectal probe electrostimulation, and evaluated. Thirteen of the males had no reproductive history. Semen characteristics (mean±SEM) recorded were as follows: semen volume, 0.91±0.11ml; semen pH, 8.1±0.1; total sperm count per ejaculate, 32.6±5.4 million; sperm viability, 84.9±1.9%; sperm motility index, 53.7±3.8; sperm abnormality, 66.1±3.4%; and percentage of sperm with a normal acrosome, 68.5±5.1%. These values were approximately in the range of the results reported in other cheetahs. These parameters, except for the percentage of sperm with a normal acrosome, were not significantly different between the fertile and subfertile males. The acrosomal integrity in the subfertile males (59.8%) was significantly lower than that in fertile ones (86.9%), but even the values in the former were not necessarily fatally low. This result indicates that their poor semen quality may not inhibit reproductive efficiency in captive male cheetahs in Japan.

View full abstract

A fatal case of bronchopneumonia associated with lungworms in a bottle-nosed dolphin, Tursiops truncatus, in an aquarium in Kochi, Kochi Prefecture, Japan, was studied. Numerous adult lungworms (Stenurus ovatus (Linstow, 1910) (Metastrongyloidea: Pseudaliidae)) were found in the bronchioles of the lungs. The nematodes as well as secondary bacterial infections may have been responsible for the development of pulmonary lesions.

View full abstract

We investigated the prevalence of pathogenic Leptospira on the small Asian mongoose (Herpestes javanicus) and the roof rat (Rattus rattus) in the northern part of Okinawa island. Leptospira were isolated from 40 (30.1%) out of 133 mongooses and 11 (20.4%) out of 54 roof rats. The strains isolated from the mongoose were identified as Leptospira interrogates serogroup Hebdomadis and Autumnalis, and Leptospira borgpetersenii serovar Javanica by microscopic agglutination test and flaB-PCR. This is the first record of serovar Javanica from mongoose inhabiting Okinawa Island. The strains from the roof rats were identified as serovar Javanica. The incidence of infection increased with the mongooses body length, which is a good index of their age. We found that in the northern part of Okinawa island, the mongoose and the roof rat are reservoirs of Leptospira, and so these animals pose a danger to people in those area.

View full abstract

The objectives of this study were to examine the utility of various body parts as a sample for sexing birds by PCR, and to establish the optimum PCR conditions and primer sets for sexing various bird species. A total of 218 samples consisting of blood, quill, rachis, down, pectoral muscle, decayed organ, skin, and crusta from 45 different bird species in captivity and the wild were collected. In 181 samples (31 of the 45 species), individual males and females were available. Optimum PCR conditions and primer sets in their respective species were established for sex identification, and amplified sex specific genes on W-chromosome and control gene on W/Z-chromosome were separated by electrophoresis. All of the blood samples and 98% of the quill samples studied were useful for sexing birds by PCR. Of the 181 samples, 175 samples were sexually determined. Sex of the individuals in 7 species was estimated, although no decisive sexing could be done because sexually known individuals were limited to either male or female. In the other 7 species, sex could not be determined because all the samples from the same species showed only a male or female electrophoresis profile by PCR. In conclusion, quill was shown to be a useful specimen for this PCR protocol, which is a practical method for sex identification of a variety of avian species. This technique has the potential to promote breeding programs in captive and wild birds.

View full abstract

To monitor reproductive status in female and male addax (Addax nasomaculatus) non-invasively, fecal reproductive steroid hormones were evaluated. Feces and blood were collected from three females and one male. Fecal contents and serum concentrations in progesterone (P_4) of the females and testosterone (T) of the male were determined by enzyme immunoassays. The changes in fecal P_4 contents during non-pregnancy and pregnancy showed the same tendency to changes in serum P_4 concentrations. There was a significant positive correlation between fecal and serum P_4 values (r=0.77, P<0.01). However, the profile of fecal T contents did not reflect the changes of the serum T concentrations in the male. A sequence of sexual behaviors as tenacious tracking, courtship and mounting by the male were observed around the time when P_4 values were basal in the female. Estrous cycle length was 28.4±0.4 days (n=5) based on the P_4 profiles. Fecal and serum P_4 values continued to rise gradually during the pregnancy. Fecal P_4 contents had reached a peak one week before parturition and then decreased rapidly. Fecal P_4 reached the basal level 3-5 days after parturition. Estrous cycles in one female recurred about 2 weeks after parturition, whereas in another female recurred about 3 months after parturition. Estrus recurrence in the latter female occurred immediately after cohabiting with a male. It was suggested that the fecal P_4 assay was quite a useful technique to determine the estrous cycle, evaluate the ovarian activity and diagnose or monitor the pregnancy.

View full abstract

Adenocarcinoma was found in a Dalmatian pelican (Pelecanus crispus) aged >25 years. A large tumor mass, which was located at the left side of the head, replaced the dura mater, skull and subcutis, and compressed the left hemisphere. In addition, neoplastic lesions were detected in the spleen, lungs and liver. Histologically, the neoplastic cells formed tubular or solid alveolar structures, and appeared to be cytologically malignant. Neoplastic cells reacting to follicle-stimulating hormone, thyroid-stimulating hormone, adrenocorticotrophic hormone, α-melanocyte-stimulating hormone, or growth hormone were demonstrated immunohistochemically. This case is presumably a primary pituitary adenocarcinoma, but the possibility of ectopic pituitary carcinoma can not be absolutely denied.

View full abstract