Influence of truck-transportation on the function of bronchoalveolar lavage fluid cells in cattle

The study sought to evaluate whether truck-transportation had an impact on the respiratory immune system of cattle. Six castrated 6-10-month-old Holstein calves were shipped approximately 100 km by road for 4 h. Plasma and bronchoalveolar lavage (BAL) fluid samples, collected immediately before transportation, at 4 h (soon after transportation), and on days 3 and 7 after transportation, were examined. A marked elevation of plasma cortisol concentration was observed at 4 h, but this level was unchanged in controls. The chemiluminescence (CL) response of phagocytes in BAL fluid cells, composed mainly of alveolar macrophage, decreased significantly after transportation (P<0.05). Transportation increased the CD3+ T cell population significantly (P<0.05), and a significant increase (P<0.05) in the ratio of CD4+ to CD8+ cells in BAL fluid was evident. We conclude that short-term road transportation alters pulmonary cells and their function, which may engender bovine respiratory disorders.     

Nucleocapsid-like structures of Ebola virus reconstructed using electron tomography

Electron tomography (ET) is a new technique for high resolution, three-dimensional (3D) reconstruction of pleiomorphic macromolecular complexes, such as virus components. By employing this technique, we resolved the 3D structure of Ebola virus nucleocapsid-like (NC-like) structures in the cytoplasm of cells expressing NP, VP24, and VP35: the minimum components required to form these NC-like structures. Reconstruction of these tubular NC-like structures of Ebola virus showed them to be composed of left-handed helices spaced at short intervals, which is structurally consistent with other non-segmented negative-strand RNA viruses.     

Effect of the well of the well (WOW) system on in vitro culture for porcine embryos after intracytoplasmic sperm injection

For developmental competence of porcine embryos in vitro, it is important to improve the culture environment. The present study was performed to evaluate four different culture systems for in vitro matured porcine oocytes following intracytoplasmic sperm injection (ICSI); drop, well and two sizes of the well of the well (WOW) systems (500 and 1,000 microm in diameter). The cleavage rate on Day 2 and the mean cell number in blastocysts on Day 6 were not significantly different among the four treatments. However, the 1,000 microm WOW (24.6%) resulted in a significantly higher (P<0.05) blastocyst rate than those in the other culture systems (12.9, 14.8, and 7.1% for drop, well, and 500 microm WOW, respectively). The present study indicates that the microenvironment created by the 1,000 microm diameter WOW improves blastocyst production of in vitro matured porcine oocytes after ICSI, and that the effectiveness of the WOW system is dependent on the size (diameter) of the WOW.     

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.     

Acceleration of follicular development by administration of vascular endothelial growth factor in cycling female rats

To address the role of follicular angiogenesis in the determination of ovulatory follicles and the effects of different vascular endothelial growth factor (VEGF) isoforms on follicular angiogenesis and development, mature female rats were treated with an angiogenic inhibitor (TNP-470), and also with VEGF 120 or 164 at different dosages (0.4, 0.8, 4.0 or 8.0 microg/kg body weight) for 3 days during the estrous cycle. Ovarian follicular angiogenesis, the population of large follicles and ovulation were examined. VEGF 120 (0.8 microg/kg) and 164 (8.0 microg/kg) treatments stimulated follicular angiogenesis in the theca interna layer, while TNP-470 treatment showed severe depression of follicular angiogenesis, and completely inhibited ovulation. After administration of VEGF 120 or 164, the number of healthy preovulatory follicles and ovulated oocytes increased significantly, concomitantly with a decrease in the number of atretic preovulatory follicles. The oocytes ovulated had normal fertilizability and developed to term with the same litter size as in the control rats. Our findings suggest that follicular angiogenesis may be a determinant of follicular development during the periovulatory phase, and that VEGF isoforms may play different important roles in regulating follicular angiogenesis.     

Expression of nerve growth factor in itchy skins of atopic NC/NgaTnd mice

Although the possible involvement of neurotrophic factors in itchy skins of atopic dermatitis has been predicted, the exact mechanism by which itch is induced remains unclear. Since nerve growth factor (NGF) has crucial effects on development and functions of sensory nerves, we determined production of NGF and extension of nerve fibers in skins of NC/NgaTnd mice with or without atopic dermatitis. NC/NgaTnd mice spontaneously develop atopic dermatitis-like skin lesions when they are raised in air-unregulated conventional circumstances. We quantified scratching behavior of NC/NgaTnd mice during the development of dermatitis using a novel analytical system and compared to clinical skin severity scores. A significant correlation between the severity of dermatitis and the increase in the number of scratches was identified, indicating that scratching behavior may associate with clinical skin conditions. NGF contents in the skin lesions of conventional NC/NgaTnd mice were significantly higher than those in SPF mice. Positive reactions for NGF were observed in keratinocytes and fibroblasts in affected skins of conventional NC/NgaTnd mice. Immunohistochemical analysis showed the extension of protein gene product 9.5-positive nerve fibers from the dermis toward the epidermis at the skin lesions. These results suggest that sensory nerves induced by NGF may contribute to development of itch, and that NGF produced at the affected site may provide abnormal skin sensitivity in atopic dermatitis.     

Gastric adenocarcinoma with ossification in a ferret (Mustela putorius furo)

A 6-year-old female ferret had a firm mass 2 cm in diameter in the pyloric region of the stomach. Histopathologically, the mass was composed of neoplastic proliferation of well-differentiated epithelial cells, showing tubular or glandular growth patterns. Osseous metaplastic foci were often found in the tumor. Tumor cells showed a positive reaction for immunohistochemistry against bone morphogenetic protein-6, an osteogenic factor. A diagnosis of gastric adenocarcinoma with ossification was made.     

Recovery of the everted cystacanth of seven acanthocephalan species of birds from feral raccoons (Procyon lotor) in Japan

Acanthocephalan everted cystacanths were detected in the intestines of 61 out of 555 feral raccoons (Procyon lotor) trapped between May 2003 and April 2005 in the western region of mainland Japan (Honshu). All collected specimens were identified as the species of birds including 3 Centrorhynchus species (C. bazaleticus, C. elongatus, and C. teres), Sphaerirostris lanceoides, Plagiorhynchus ogatai, Porrorchis oti, and Southwelina hispida. Three species, C. bazaleticus, C. teres, and S. lanceoides, were new to distribution records in Japan and the Far East. Recovery rates of these acanthocephalans were higher from the raccoons trapped in spring and early summer (range 8.3-36.8%, average 21.5%) than during the remainder of the year (range 0-20.0%, average 6.3%). The reason for this seasonal change is unknown. There is little information on the acanthocephalan fauna of wild birds in Japan, however, recovery of the cystacanths from the feral raccoons captured through the year might provide useful information on the fauna and geographical distribution of avian acanthocephalans.     

Experimental dual infection of pigs with an H1N1 swine influenza virus (A/Sw/Hok/2/81) and Mycoplasma hyopneumoniae

Dual infection of pigs with swine influenza virus (SIV) and Mycoplasma hyopneumoniae was carried out to compare the clinical and pathological effects of dual infection in caesarian derived and colostrums deprived (CDCD) pigs, with that of a single infection with M. hyopneumoniae. In Experiment 1, 40-day-old CDCD pigs were inoculated only with SIV (A/Sw/Hok/2/81, H1N1). The virus was isolated from nasal swabs for 5-6 days. None of these pigs showed clinical signs of infection throughout the experimental period. These results suggested that this strain can infect pigs but is only slightly pathogenic when it is inoculated singly to a CDCD pig. In Experiment 2, 60-day-old CDCD pigs were inoculated with M. hyopneumoniae and then were inoculated with SIV (A/Sw/Hok/2/81) at 1 week (MHYO-7d-SIV-7d group) or 3 weeks (MHYO-21d-SIV-7d group) after M. hyopneumoniae inoculation. Macroscopically, dark red-to-purple lung lesions were observed in all of pigs at 14 or 28 days post-inoculation. Percentages of dark red-to-purple lung lesions in dual infection groups (MHYO-7d-SIV-7d group: 18.7 +/- 4.2%, MHYO-21d-SIV-7d group: 23.0 +/- 8.0%) were significantly (P < 0.05) increased compared to those of each control group in which pigs were inoculated only with M. hyopneumoniae (MHYO-14d group: 4.7 +/- 2.9%, MHYO-28 group: 3.3 +/- 2.4%). Microscopically, bronchial epithelial lesions (epithelial disruption, degeneration, hyperplasia and formation of microabscess) were frequently observed in dark red-to-purple lung lesions of only the dual infection groups. These results demonstrate that the lung lesion of pigs inoculated with M. hyopneumoniae and SIV is more severe than that of pigs inoculated only with M. hyopneumoniae.     

Cloning of bovine MAIL and its mRNA expression in white blood cells of Holstein cows

Molecule possessing ankyrin-repeats induced by lipopolysaccharide (MAIL) is known as an IkappaB protein induced after administration of bacterial lipopolysaccharide (LPS) to mice. In the present study, we cloned bovine MAIL cDNA and examined its mRNA expression in white blood cells isolated from Holstein cows. Bovine MAIL had more than 80% amino acid identities with murine and human MAILs, highly conserved ankyrin-repeat motifs and PEST-like sequences. Bovine MAIL mRNA was undetectable in isolated peripheral white blood cells, but rapidly induced (<1h) after stimulation by LPS and lipid A in vitro in a dose-dependent manner. The lipid A-induced MAIL mRNA expression was found in polymorphonuclear cells, monocytes/macrophages and total lymphocytes, but not in T-lymphocytes. MAIL mRNA was also induced in vivo in peripheral blood leukocytes of cows after intramammary injection of Escherichia coli derived from coliform mastitis. Thus, bovine MAIL, as rodent MAILs, is induced by inflammatory stimuli in specific immune cells in vitro and in vivo, suggesting a role in inflammatory responses to bacterial infection in cattle.