Characterization of mutations in the two-component histidine kinase gene that confer fludioxonil resistance and osmotic sensitivity in the os-1 mutants of Neurospora crassa

Osmotic-sensitive (os-1) mutant alleles in Neurospora crassa exhibit resistance to dicarboximides, aromatic hydrocarbons and phenylpyrroles. We have previously reported that the os-1 mutants can be classified into two groups based on their resistance to fungicides and osmotic stress: type I, which are highly resistant to iprodione and fludioxonil but moderately sensitive to osmotic stress, and type II, which are highly sensitive to osmotic stress but moderately resistant to fungicides. To explain the mechanism of resistance to these fungicides, we cloned and sequenced the mutant os-1 genes that encode putative osmo-sensing histidine kinase. Within the os-1 gene product (Os1p), the type I strains, NM233t and Y256M209, carried a stop codon at amino acid position 308 and a frameshift at amino acid position 294, respectively. These mutation sites were located on the upstream of histidine kinase and the response regulator domains of Os1p, strongly suggesting that type I strains are null mutants. The null mutants, NM233t and Y256M209, were highly resistant to iprodione and fludioxonil; thus Os1p is essential for these fungicides to express their antifungal activity. The amino acid changes in Os1p, 625Pro from Leu, 578Val from Ala, and 580Arg from Gly were found in the type II strains, M16, M155-1 and P5990, respectively. Os1p is novel in having six tandem repeats of 90 amino acids in the N terminal. Each amino acid change of the type II strains was located on the fifth unit of six tandem repeats. Type II strains with single amino acid changes were more sensitive to osmotic stress than the null mutants (type I), indicating that the amino acid repeats of Os1p were responsible for an important function in osmo-regulation.     

Peripheral neuroblastoma in a young labrador retriever

A 2-year-old Labrador Retriever developed atrophy of the right temporal muscle, subsequently showed generalized seizure and died 2 months after the clinical onset. Postmortem examination revealed the tumor masses in the right mandibulopharyngeal area, nasopharynx and intracranial space. Histopathologically, these tumor masses were composed of small round neoplastic cells and neuropil-like stroma separated by fibrovascular septa. In the neoplastic masses, small neoplastic cells with round to oval hyperchromatic nuclei and scanty cytoplasm predominated, and angulated neoplastic cells with larger nuclei and moderate cytoplasm were scattered. Immunohistochemically, neoplastic cells were positive for neuron specific enorase, neurofilament protein, chromogranin A, synaptophysin and tyrosine hydroxylase. Based on these findings, this case was diagnosed as peripheral neuroblastoma, presumably originated from the sympathetic ganglion, maybe right craninal cervical ganglion.     

Age-related changes of genital systems in the female Crj:CD (SD) IGS rats during sexual maturation

The age-related changes of vaginal opening, body weight, the weights of the uterus and ovary, together with histological examination, serum 17beta-estradiol (E2) and progesterone levels were examined in intact female Crj:CD (SD) IGS rats between 21 and 36 days of age to understand the basic biological profile of changes of the female genital system during sexual maturation in the rat for female pubertal assays. With the beginning of the elevation of serum E2 level from 28 days of age, all parameters except body weight started to show drastic change until 31 days of age. The highest incidence of vaginal opening was recorded at 34 days of age. On macroscopic examinations, a number of rats showed uterine imbibition but vaginal opening. Immediately after the confirmation of the vaginal opening, the genital systems of three rats were observed microscopically. Both ovaries already had multiple corpora lutea, and degeneration of endometrial epithelial cells was observed. In conclusion, we obtained essential data on genital tract development of female Crj:CD (SD) IGS rats for in vivo screening assays that will contribute to detect potential endocrine active chemicals. In addition, it is assumed that the first ovulation precedes or occurs simultaneously with vaginal opening.     

Importance of Escherichia coli infection in ascites in broiler chickens shown by experimental production

Common commercial strain male broilers aged 14 days were intratracheally inoculated with 0.2 ml of 1.2 x 10(6) colony-forming units of Escherichia coli in nutrient broth and kept in a cool environment during the experiment. Ascites was produced in five surviving and two dead birds out of 50 but not in 50 mock-infected control birds. Among the 40 survivors that were infected, the erythrocyte packed cell volume (PCV) of the 10 birds with pericarditis was the same as in 21 grossly normal birds, although that of the four birds with enlarged right ventricle (RV) was high. The pericarditis caused by E. coli septicemia was not the primary cause of ascites. However, the PCV was high in some of the survivors with an enlarged RV without pericarditis, indicating overload due to the lung lesion. These data suggested that some of the birds with an enlarged RV, caused by supplying blood that was insufficiently oxygenated for the body size, suffered from ascites.     

An encephalopathy with argyrophilic inclusions in a Holstein-Friesian cow

The brain of a 6-year-old Holstein cow, which showed progressive neurologic symptoms during several months, was examined by histopathologic methods. Many round or oval-shaped cytoplasmic inclusions were observed, mainly in neurons of the temporal lobe and the hippocampus. Those inclusions were faintly eosinophilic with hematoxylin and eosin and positive with Bielschowsky's silver stain. Immunohistochemically, the inclusions were recognized by antiubiquitin and antiphosphorylated tau antibodies. Ultrastructurally, the inclusions were globular and well demarcated from the rest of the cytoplasm, lacked limiting membranes, and were mainly composed of straight fibrils about 15 nm in width. The structure of the inclusions was similar to that of Pick bodies in Pick's disease of humans. The pathogenesis of this bovine condition is not known.     

A pathologic study on ocular disorders in calves in southern Kyushu, Japan

Of 822 calves, ranging in age between one day and six months necropsied between 1996 and 1998 at Miyazaki University, histological examination showed that 25 (3.0%) had ocular lesions. These ocular lesions consisted of suppurative inflammation (13 cases), cataract (seven cases), and retinal atrophy (five cases). Inflammatory changes were classified as suppurative keratitis (one case), keratitis and uveitis (ten cases), and uveitis and retinitis (two cases). Cataract was subclassified into three categories; cortical (three cases), nuclear (one case), and mature (three cases). These lesions were characterized by degenerative changes in the lens fibers and the appearance of eosinophilic globules known as Morganian globules. In the most severely affected case, there was capsular rupture of the lens, resulting in severe infiltration by eosinophils and histiocytes of the whole anterior chamber. Almost all the calves with retinal atrophy had been suffering from severe hydranencephaly and three had significantly raised levels of neutralization antibodies for the Akabane and/or Aino viruses. This study indicates that congenital arbovirus infections may predispose calves to ocular diseases, especially retinal atrophy.     

An epizootiological survey of necropsy cases (1993-1997) at University of the Philippines

An epizootiological survey of necropsied cases (1993-1997) at University of the Philippines was performed. A total of 368 cases included 238 avian and 111 porcine cases. Amongst avian cases, the major cause of death was infectious diseases in 212 (89%) cases including 97 (41%) bacterial, 36 (15%) viral, and 21(9%) parasitic diseases. The majority of the avian bacterial diseases presented as septicemia (73 cases) and the viral diseases as Newcastle disease (17 cases). In porcine cases, the major cause of death was also infectious diseases, in 100 (90%) cases including 52 bacterial and 29 viral diseases. Porcine bacterial diseases were classified into 36 septicemia, 4 hemophillosis and 4 colibacillosis. Amongst the porcine viral diseases, most cases were diagnosed as Hog cholera (22 cases).     

竞争性PCR定量检测鸡传染性贫血病毒核酸

将删除33个碱基序列的CAVDNA克隆进质粒pSBII,提取质粒DNA,经过纯化、定量后作为竞争性模板,建立了一种定量检测鸡传染性贫血病毒核酸方法。该方法能够检测出的最低CAVDNA为103.5Molecules/μL,其精确度可达到100.5Molecules/μL。该方法与传统方法相比,具有节省时间、节约试剂和受其它因素干扰小等特点。     

Development and introduction of detection dogs in surveying for scats of small Indian mongoose as invasive alien species

The small Indian mongoose was introduced to the island of Okinawa in 1910 to control rats and snakes, however, it poses a threat to many endemic species in the Yambaru forest region in Okinawa Island. An important concern in eradication is that, as much as possible, capture should be carried out without affecting native species. To protect endemic species in the Yambaru region of Okinawa Island, we trained and used detection dogs to search for mongoose scats that indicated that mongoose were present in the area. We confirmed the ability of the dogs to detect scats in positive and negative control tests in the Yambaru forest region after basic, obedience, and mongoose-scat-detection specific training. In 30 trials, the dogs found 60 of 65 scats samples. For the trained alert, the dogs carried out 59 trained alerts for the 60 scats found. In the positive and negative control tests, the dogs showed no reaction to 129 of 130 negative controls. We then surveyed the forest in the Yambaru region for mongoose scats using the detection dogs. In this study, the detection dogs found 79 scats in a survey of a 69.0-km area. We divided the survey area into 43 grids and calculated the rate of detection in each grid. The rate of detection of scats was higher in the southern part of the Yambaru region. These result showed that our mongoose scat detection dogs can be useful tools to assist in determining the presence of mongoose in the Yambaru region of Okinawa Island.     

Identification of oyster-derived hypotensive peptide acting as angiotensin-I-converting enzyme inhibitor

Angiotensin-I-converting enzyme (ACE) plays a crucial role in the crisis of hypertension. Some peptides that originate from protease hydrolysates are known to suppress ACE activity in vitro and in vivo. Here, we investigated whether trypsin hydrolysate of oyster Crassostrea gigas showed hypotensive activity and ACE inhibition. The hydrolysate significantly suppressed systolic blood pressure and ACE activity in spontaneously hypertensive rats following a one-shot oral administration and a long-term feeding experiment lasting 9 weeks. Each hydrolysate from oyster tissue showed ACE inhibitory activity, indicating the hypotensive effect was due to synergism. One potent ACE inhibitory peptide, Asp-Leu-Thr-Asp-Tyr, was identified from the hydrolysate of the striate muscle, and the peptide exhibited hypotensive activity in vivo. Protease digestion analysis suggested that Asp-Tyr could be the real effector of this penta-peptide in vivo.