Establishment and characterization of eight feline mammary adenocarcinoma cell lines

Eight new feline mammary adenocarcinoma cell lines derived from either primary or metastatic lesions were established. The morphology of all the cell lines was epithelioid and round to spindle in shape, with cell growth occurring in a monolayer fashion. On immunohistochemistry, these cells reacted with anti-keratin and anti-vimentin antisera. The doubling time of these cells was between 19 and 54 hr. Tumor masses were developed in nude mice by subcutaneous inoculation of the cells that were histologically identical to their original mammary tumor lesions. Telomerase activities measured using the telomeric repeat amplification protocol assay revealed high telemetric activity in all of the cells.     

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.     

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.     

Feed intake,digestibility, nitrogen utilization,ruminal condition and blood metabolites in wethers fed ground bamboo pellets cultured with white‐rot fungus (Ceriporiopsis subvermispora) and mixed with soybean curd residue and soy sauce cake

Three types of bamboo pellets as a ruminant feed: P1 (ground bamboo (GB) cultured with the fungus Ceriporiopsis subvermispora (CGB) : soybean curd residue (T) : soy sauce cake (S) in a 5:4:1 ratio on a dry matter (DM) basis); P2 (GB : T : S = 5:4:1 on a DM basis); and P3 (CGB : T : S = 5.5:0.8:3.7 on a DM basis) were prepared. Four wethers were assigned in a 4 × 4 Latin square design experiment to evaluate the applicability of the bamboo pellets. The experimental treatments were C (control): fed alfalfa hay cubes (AC) only, and T1, T2 and T3: fed P1, P2, and P3 with AC by 1:1 on a DM basis, respectively. The digestibility of the DM, organic matter and acid detergent fiber of P1 were significantly higher than those of P2 and P3 (P < 0.05). The total digestible nutrient (TDN) contents of AC, P1, P2 and P3 were 56.5%, 60.2%, 53.2% and 47.0%, respectively. No significant differences in nitrogen retention or ruminal pH and NH₃ were observed among the treatment groups. The results indicate that bamboo pellets cultured with C. subvermispora and mainly mixed with soybean curd residue improved nutritional quality of ground bamboo because of its high digestibility and TDN content.     

Identification of bovine serum albumin as an IgE-reactive beef component in a dog with food hypersensitivity against beef

IgE-reactive beef components were examined by an immunoblot analysis using a serum from a dog with food hypersensitivity against beef. The immunoblot analysis revealed a distinct band at approximately 66 kDa and a faint band at approximately 50 kDa. The immunoblot analysis for serum IgE reactivity to bovine serum albumin (BSA) also revealed a positive band at 66 kDa. Serum IgE reactivity to the 66-kDa protein of beef was diminished by pre-incubating the serum sample with BSA. Furthermore, a positive reaction to BSA was detected in intradermal testing in the dog. These results clearly indicated that BSA was an IgE-reactive beef component in the dog with food hypersensitivity against beef.     

Increased plasma levels of soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular cell molecule-1 (sVCAM-1) associated with disease severity in a primate model for severe human malaria: Plasmodium coatneyi-Infected Japanese macaques (Macaca fuscata)

In the present study, we investigated plasma levels of soluble intercellular adhesion molecule-1 (sICAM-1) and vascular cell adhesion molecule-1 (sVCAM-1) in seven Japanese macaques (Macaca fuscata) infected with Plasmodium coatneyi. Concentrations of sICAM-1 and sVCAM-1 were significantly elevated in the severe phase; the levels were maximally increased up to six times and three times those before infection, respectively. We subsequently examined kinetic profiles of sICAM-1 and sVCAM-1 concentration in plasma obtained from two infected monkeys. Both infected monkeys had markedly increased levels of these adhesion molecules when they exhibited severe clinical signs correlated with rapid increase in parasitemia. These results suggest that the elevation of levels of sICAM-1 and sVCAM-1 is a critical step in the pathogenesis of severe malaria in vivo.     

Epidemiological and bacteriological survey of buffalo mastitis in Nepal

A total of 355 Murrah cross buffaloes, consisting of 23 subclinical and 332 clinical mastitis cases brought to the Veterinary Teaching Hospital, Chitwan, Nepal from 2002 to 2005, were analyzed to determine the organisms involved, the seasonal occurrence of mastitis, and antibiotic susceptibility of mastitis pathogens. Coagulase negative Staphylococci (CNS) such as Staphylococcus albus and S. epidermidis were the predominant organisms associated with subclinical cases, and CNS and Coliforms in clinical cases. The maximum number (16%) of clinical cases of mastitis were observed in the month of July, when temperature and humidity are highest. The incidence of clinical mastitis was higher in animals during 1st calving and during the 1st month of parturition. Resistance to antibiotics was determined for 55, 23 and 149 isolates of Staphylococcus spp., Streptococcus spp. and Coliforms, respectively. In vitro drug sensitivity testing revealed that enrofloxacin had the highest average sensitivity (91%) for all types of bacteria. The effectiveness of other drugs detected were gentamicin (87%), tetracycline (83%) and chloramphenicol (82%). The antibiogram showed that both gentamicin and enrofloxacin are slowly becoming resistant. Mastitis pathogens have developed resistance to ampicillin and penicillin.     

Clinical response of ovarian cysts in dairy cows after PRID treatment

We investigated the therapeutic effects of a progesterone releasing intravaginal device (PRID) on cystic ovarian disease (COD) and reproduction performance of cows. The possible influence of PRID on metabolic and/or health status was also examined. A total of 40 Holstein-Friesian cattle, with ovarian cystic structures, > or =2.5 cm in diameter, persisting for more than 7-14 days, without a corpus luteum (CL) were used for the study. PRID or placebos were inserted into the vagina for 12 days. Five animals lost the intravaginal device before removal and one was culled. Based on plasma progesterone concentration on the day of treatment, 20 (17 PRID and 3 placebos) of the remaining 34 cows had follicular cysts (progesterone < or =1 ng/ml) and 14 (10 PRID and 4 placebos) had luteal cysts (progesterone >1 ng/m l). Fourteen (82%) of the PRID-treated follicular cystic cows responded with formation of a CL within 14 days after treatment, and an overall conception rate of 53.8%. Likewise, 70% of the treated luteal cystic cows responded with CL formation and 71.4% conception rate. No significant differences were observed in hematocrit (Ht), white blood cell count and serum levels of glucose, blood urea nitrogen, aspartate aminotransferase, and alanine aminotransferase, between the day of PRID insertion and removal, in animals with follicular and luteal cysts. PRID treatment resulted in ovulation 2-4 days later and formation of a CL in cows that recovered.