A pathogenic Entamoeba histolytica-like variant (JSK2004 strain) with genetic variations and a novel isoenzyme pattern isolated from a De Brazza's guenon in a Tokyo zoo in Japan has previously been documented. In this study, a multiplex polymerase chain reaction (PCR) assay that could distinguish the JSK2004-type E. histolytica-like variant (JSK04-Eh-V) from E. histolytica and Entamoeba dispar using three newly designed primer sets for amplifying each specific DNA fragment from their small-subunit ribosomal RNA genes was developed and established. Forty-seven primates (11 species) from the zoo were surveyed by multiplex PCR to assess the prevalence of JSK04-Eh-V infection, which was recognized in six individuals of four species, including an Abyssinian colobus monkey, a De Brazza's guenon (including the individual from whom JSK2004 was isolated), a white-faced saki, and a Geoffroy's spider monkey. In addition, the autopsied individuals of an Abyssinian colobus and Geoffroy's spider monkey that died of amoebic liver abscess were also evaluated. DNA samples were also analyzed for specific genotypes based on the nucleotide sequencing of two protein-coding (chitinase and serine-rich E. histolytica protein) genes and the protein-noncoding locus 1-2 that was used for fingerprinting of the E. histolytica strain. These studies indicated that the E. histolytica-like variant infection in this zoo was caused by the same type (i.e., JSK04-Eh-V). An axenic culture medium (yeast extract-iron-maltose-dihydroxyacetone-serum) was developed based on the yeast extract-iron-gluconic acid-dihydroxyacetone-serum medium, which is designed for axenic culture of E. dispar. This new medium could be used for axenically culturing E. histolytica, JSK04-Eh-V, and E. dispar in a single medium.
ABSTRACT. Avian tuberculosis accompanied with many tubercular lesions in the liver and spleen was found in a painted quail at a zoological garden in Japan. Mycobacterium avium complex (MAC) serovar 9 without insertion sequence of IS901 was isolated from the liver (1.3 × 10 8 CFU/g), oviduct (9.4 × 10 7 CFU/g), and intestine (1.5 × 10 5 CFU/g). The isolates were inoculated intravenously to chickens. The inoculated chickens showed clinical symptoms of avian tuberculosis. Birds are susceptible to MAC serovar 9 without IS901.-KEY WORDS: avian tuberculosis, Mycobacterium avium complex serovar 9, painted quail.
ABSTRACT. An intrauterine insemination technique using a fiberscope was investigated in the giant panda. A septal wall was present about 5 cm from the vulva, the external urethral orifice was present in the ventral region, and the entrance (pseudocervix) to the vagina was present in the dorsal region. The uterovaginal region protruded in the dorsal region about 15 cm from the pseudocervix. The external uterine orifice was present in the uterovaginal region, revealing that intrauterine insemination can be easily performed. This technique may greatly contribute to artificial reproduction of the giant panda. KEY WORDS: fiberscope, giant panda, intrauterine insemination.J. Vet. Med. Sci. 68(9): 987-990, 2006 The giant panda (Ailuropoda melanoleuca: G. panda) is an endangered species, and many efforts are being made to conserve this animal. Ueno Zoo (Tokyo, Japan) started a raising and reproduction program for G. pandas in 1972, and 3 births were obtained by AI in 1985AI in , 1986AI in , and 1988. All were obtained by intravaginal insemination using bovine insemination equipment. However, no conceptions have been achieved since then.Artificial insemination (AI) is divided into intrauterine and intravaginal based on the site of semen infusion, and the number of sperm required to obtain conception by these methods is markedly different. For example, in beagles, 2 × 10 8 sperms are required for intravaginal insemination, but only 1/10 of this, or 2 × 10 7 sperms, is required for intrauterine insemination [14]. The number of sperm required for intrauterine insemination is 1/5 of that required for intravaginal insemination in cats [11,13]. In frozen semen, sperm motility markedly decreases and survival time shortens after thawing [8][9][10]. For efficient usage of sperm in an endangered species, establishment of an effective intrauterine insemination technique is urgently needed.There is very little information on the intravaginal structure of living G. pandas. A narrow region is present about 5 cm from the vulva, and the vagina is located in the inner part, but it cannot be visually confirmed because of the body shape of the animal. Development of intrauterine insemination using a laparoscope is underway for animals in which transvaginal intrauterine insemination is not applicable [4]. However, this method requires an incision in the abdominal wall and takes time for insemination. Thus, development of a non-invasive intrauterine insemination technique is necessary for rare animals including the G. panda.The objective of this study was to clarify the fiberscopic anatomical characteristics of the vaginal cavity, which is the region from the vulva to the external uterine orifice, and investigate the development of a transvaginal intrauterine insemination technique using a fiberscope for efficient usage of G. panda sperm.Animals: A female G. panda that had been maintained in Mexico (Chpultepec Zoo) was transferred to Ueno Zoo for AI. The animal was 16 years of age, and 130 kg in weight. Semen was collected from a 19-yea...
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