Molecular breeding for virus resistant potato plants

To engineer resistance against potato virus X (PVX), the viral coat protein (CP) gene has been introduced into two potato cultivars. Stable expression of the gene in transgenic clones throughout the growing season has been obtained and resulted in considerably increased virus resistance. With varying frequencies depending on the original cultivar used, true-to-type PVX resistant transgenic clones have been obtained. Since deviant light sprout characteristics were invariably associated with aberrations in plant phenotype, they can be used in procedures to early screen for deviations. Furthermore, it has been possible to unequivocally discriminate between the original untransformed and independent transgenic cultivars. Although no relation has been found between the presence, if any, of the CP of potato virus Y (PVY) or potato leafroll virus (PLRV) in CP gene transgenic potato, appreciable levels of resistance to these viruses has been obtained. This suggests that the mechanism by which a viral CP gene in the potato genome evokes resistance, differs amongst various viruses.     

Molecular heterogeneity in clinical isolates of Malassezia pachydermatis from dogs

Molecular investigation of 16 strains, conventionally identified to be Malassezia pachydermatis, isolated from dogs in Japan was carried out by random amplification of polymorphic DNA (RAPD) and chitin synthase 2 (CHS2) gene sequence analyses. The RAPD band patterns of 13 clinical isolates were identical to that of standard strain of M. pachydermatis (CBS-1879). The other three clinical isolates were different from the standard strain of M. pachydermatis in RAPD patterns, and two of the three isolates were identical. About 620 bp genomic DNA fragments of the CHS2 gene were amplified from the same 16 clinical isolates of M. pachydermatis by polymerase chain reaction (PCR) and sequenced. The phylogenetic analysis of the nucleotide sequences of CHS2 gene fragments of the 16 clinical isolates revealed that the 13 strains were genetically very close to the standard strain of M. pachydermatis and the other two isolates were genetically close to the standard strain of M. furfur rather than M. pachydermatis. The remaining one isolate was phylogenetically distinct from all the seven Malassezia species reported so far.     

Shifts in disease resistance induced by growth regulators

Experiments on the effect of various compounds with plant growth regulating activity led to the hypothesis that conditions inhibitory to indoleacetic acid (IAA) action or leading to a decrease in the IAA level in cucumber seedlings would be unfavourable for the development of cucumber scab, caused byCladosporium cucumerinum. Susceptibility decreased as the result of treatment with growth retardants, which cause an increase in IAA-oxidase activity in the plants, whereas application of indoleacetic acid or compounds expected to decrease the rate of oxidative breakdown of IAA in the seedlings resulted in an increase of susceptibility. Growing the plants under various periods of illumination also influenced both the susceptibility of the plants and the IAA-oxidase activity in the hypocotyl tissue.To obtain further information about the relation between the increase in resistance and the increase in the rate of IAA oxidation, the effect of one compound, viz. L-threo-β-phenylserine (abbr. phenylserine) was studied in more detail. The rate of oxidative breakdown of IAA was increased in extracts of hypocotyls and cotyledons of plants treated with phenylserine as compared with extracts of tissue of untreated plants. Extracts of phenylserine-treated plants were found to contain a higher cofactor and/or a lower inhibitor content than those of control plants. The substances involved were not identified, but this result may indicate a shift in the concentrations of various phenols in the tissue.The question must remain open whether a shift in the concentration of phenols some-how effected by phenylserine treatment determines both degree of susceptibility and rate of IAA oxidation independently, or whether the rate of IAA breakdown and the resulting change in IAA content in the plants is directly related to the degree of susceptibility.     

Cerebral high-grade astrocytoma (glioblastoma) in a cat

A cerebral tumour was found in the right frontal lobe of a 7-year-old female mongrel cat. The mass showed infiltrative growth and caused deformation of the corpus callosum. Histopathologically, the tumour cells showed anaplasia, pleomorphism and mitotic figures. Necrosis and vascular proliferation were prominent. The neoplastic cells surrounded areas of necrosis, but as an indistinct pseudopalisade formation. Immunohistochemically, low numbers of tumour cells labelled positively for anti-glial fibrillary acidic protein and anti-S100 protein. Electron microscopically, the majority of tumour cells had no filaments and cytoplasmic processes, but the differentiated cells presented cytoplasmic filaments and glycogen granules. Based on these findings, the tumour was diagnosed as cerebral high-grade astrocytoma, glioblastoma.     

Protective effects of sugar cane extracts (SCE) on Eimeria tenella infection in chickens

The effects of oral administration of sugar cane extracts (SCE) on Eimeria tenella oocysts infection in chickens were studied with 2 different experiments. In Experiment 1, 3-week-old inbred chickens (MHC; H.B15) were inoculated into the crop with SCE (500 mg/kg/day) for 1 day or 3 consecutive days, and then challenged with E. tenella sporulated oocysts (2 x 10(4) cells/chicken). In Experiment 2, 1-week-old chickens were orally administered SCE at the same dose for 3 consecutive days, and then initially infected with E. tenella sporulated oocysts (2 x 10(3) cells/chicken). At 2 and 3 weeks of age, these chickens were immunized intravenously with the mixed antigens of sheep red blood cells (SRBC) and Brucella abortus (BA). At 4 weeks of age, chickens were challenged with E. tenella sporulated oocysts (1 x 10(5)/chicken). Challenged chickens with E. tenella oocysts showed markedly decreased body weight gain/day, severe hemorrhage and great number of shedding oocysts in feces and high lesion scores. Oral administration of SCE and initial infection with oocysts (2 x 10 (3)/chicken) resulted in a remarkable improvement in body weight gain/day, hemorrhage, the number of shedding oocysts and lesion score, compare to other infected groups. In addition, SCE-inoculated chickens with the initial infection showed a significant increase in antibody responses against SRBC and BA and also improvement in decreased relative proportions of Bu-1a(+) and CD4( )cells in cecal tonsil lymphocytes of E. tenella-challenged chickens. Cecal tissues of chickens administered SCE and initially infected with E. tenella oocysts showed lower numbers of schizonts, gametocytes and oocysts than those of infected control chickens. These results suggest that SCE have immunostimulating and protective effects against E. tenella infection in chickens.     

New potyvirus isolated from <Emphasis Type="Italic">Cryptotaenia japonica</Emphasis>

 A potyvirus, for which the name Japanese hornwort mosaic virus (JHMV) is proposed, was isolated from Japanese hornwort plants (Cryptotaenia japonica) with mosaic disease symptoms. The virus was used to inoculate mechanically 34 plants belonging to 33 species of 10 families. Of these species seven from two families were infected. Faint chlorotic spots appeared on the inoculated leaves of Chenopodium quinoa and C. amaranticolor, but no systemic infection occurred in these plants. JHMV systemically infected only Umbelliferae plants; they did not infect 26 other species in eight families. JHMV was transmitted in a nonpersistent manner by aphids (Myzus persicae). The virus was a flexuous rod-shaped particle about 750 nm in length. Sequencing the nucleotides in the 3′ terminal region of JHMV revealed that the coat protein contains 280 amino acids with a molecular mass of 32.2 kDa. The nucleotide sequence of the coat protein of JHMV had the highest similarity with that of Zantedeschia mosaic virus (83.3%) compared to those of other potyviruses (57.0%–64.9%). An antiserum against JHMV reacted strongly with JHMV and weakly with Potato virus Y. These results indicate that JHMV is a new potyvirus. Received: September 9, 2002 / Accepted: November 7, 2002 RID="*" ID="*" The nucleotide sequence determined in this work appears in the DDBJ/EMBL/GenBank nucleotide sequence databases with the accession number AB081518     

Decrease in malate dehydrogenase activities in peripheral leucocytes of type 1 diabetic dogs

The activities of the enzymes involved in the malate-aspartate shuttle were measured in peripheral leucocytes of dogs with type 1 diabetes mellitus. In the diabetic dogs, fasting plasma glucose concentrations were twofold greater than control levels despite insulin injections and the activities of malate dehydrogenase (MDH), which plays a crucial role in the malate-aspartate shuttle, were decreased remarkably. The cytosolic ratio of MDH/lactate dehydrogenase (LDH) activity (M/L ratio) in leucocytes of the diabetic dogs was significantly lower than that of normal control dogs. The decrease of the M/L ratio appeared to reflect depression of energy metabolism in leucocytes of the diabetic dogs. The M/L ratio may be a useful parameter to evaluate metabolic conditions in diabetic dogs.     

Electrocorticographic and histological findings in a Shetland sheepdog with intractable epilepsy

A Shetland sheepdog with epilepsy refractory to antiepileptic drugs was brought to the division of Veterinary Radiology at Nippon Veterinary and Animal Science University. Scalp electroencephalography and computed tomography was performed, but no abnormality was detected in either examination. To obtain detailed information, electrodes were implanted on the dura mater, and the electrocorticogram (ECoG) was recorded. In the ECoG, sporadic spikes were detected in the left parietal region, suggesting the presence of the epileptic focus in this region. After the dog's death, abnormalities of gyri were found in the region where spikes were detected in the ECoG. On histopathological examination, laminar malacia of the cingulate gyrus was observed. Furthermore, in the hippocampus, neuronal loss of pyramidal cells was observed.     

Classification of abomasal displacement in cows according to histopathology of the liver and clinical chemistry

Histopathological features of livers and blood chemical values in cows with abomasal displacement were investigated. Liver biopsy samples were collected during redressment operations in 92 cows with abomasal displacement, and the samples were stained with haematoxylin and eosin or periodic acid Schiff (PAS). Blood was collected for chemical tests. Livers were histopathologically divided into the following four types: normal histology cases (21%), fatty degeneration cases (36%), cloudy swelling cases (19%) and fatty degeneration cases with cloudy swelling (24%). The number of PAS-positive samples was significantly higher in the normal histology group and significantly lower in the severe fatty degeneration group and severe cloudy swelling group. Cows with fatty degeneration had significantly higher levels of serum 3-hydroxybutyric acid, non-esterified fatty acid and aspartate aminotransferase than did those with cloudy swelling or normal histology. The results indicate that the morbid conditions of cows with abomasal displacement can be classified into four types.     

Hypolipemia associated with the wasting condition of rabbits infected with Strongyloides papillosus

Rabbits develop a wasting condition in the intestinal stage of Strongyloides papillosus infection. Serum inflammatory cytokine and lipid profiles were investigated in five rabbits infected with S. papillosus and five uninfected pair-fed controls to ascertain whether the disease is inflammatory cytokine-mediated cachexia. Tumor necrosis factor alpha (TNF alpha) was detected in one infected animal at Day 7 after infection. Interleukin (IL)-1 was detected in three infected, and one control, animals at Day 28. IL-6 remained unchanged in both the groups. Infected animals developed hypolipemia, including hypotriglyceridemia in the intestinal stage of infection. Control animals lost body weight in the same manner as the infected animals, but had elevated cholesterols and phospholipids with normal triglyceride concentrations. The results suggested that the wasting condition has no association with cachexia induced by TNF alpha. IL-1 or IL-6, and that hepatic function for lipid synthesis is affected during the intestinal stage of S. papillosus infection.