Antitumor effect of adriamycin entrapped in liposomes conjugated with anti-bovine tumor antigen monoclonal antibody in leukemic cows

Monoclonal antibody c143 against tumor-associated antigen (TAA) expressed on bovine leukemia cells was conjugated to liposomes containing adriamycin (ADM), and therapeutic effects of conjugates were examined in leukemic or preleukemic cows to prevent the progression of the disease. Five cows with TAA-positive in their peripheral blood lymphocytes were divided into two groups. Each group of cows received 4 injections of ADM alone (0.4 mg/kg) or c143-conjugated liposomes containing the same dose of ADM (L-AMD-c143) through the jugular vein at about 4 day intervals. In three animals treated with L-ADM-c143, the TAA-positive cells gradually decreased with treatment and finally two animals became TAA-negative during a 6 week period and a 14 week period after treatment, respectively. In the control, two animals treated with ADM alone showed only a slight decrease of TAA-positive cells.     

Catalog of Micro-Tom tomato responses to common fungal, bacterial, and viral pathogens

Lycopersicon esculentum cultivar Micro-Tom is a miniature tomato with many advantages for studies of the molecular biology and physiology of plants. To evaluate the suitability of Micro-Tom as a host plant for the study of pathogenesis, Micro-Tom plants were inoculated with 16 well-known fungal, bacterial, and viral pathogens of tomato. Athelia rolfsii, Botryotinia fuckeliana, Oidium sp., Phytophthora infestans, and Sclerotinia sclerotiorum caused typical symptoms and sporulated abundantly on Micro-Tom. Micro-Tom was resistant to Alternaria alternata, Corynespora cassiicola, and Fusarium oxysporum. When Micro-Tom was inoculated with 17 isolates of Ralstonia solanacearum, many isolates induced wilt symptoms. Agrobacterium tumefaciens also was pathogenic, causing crown galls on stem tissue after needle prick inoculation. In Micro-Tom sprayed with Pseudomonas syringae pv. tomato, P. s. pv. tabaci, or P. s. pv. glycinea, bacterial populations did not increase, and yellow lesions appeared only on leaves sprayed with P. s. pv. tomato. Tomato mosaic virus, Tomato aspermy virus, and Cucumber mosaic virus systemically infected Micro-Tom, which developed symptoms characteristic of other cultivars of tomato after infection with the respective virus. These results indicated that Micro-Tom was generally susceptible to most of the important tomato pathogens and developed typical symptoms, whereas certain pathogens were restricted by either hypersensitive resistance or nonhost resistance on Micro-Tom. Therefore, an assortment of Micro-Tom–pathogen systems should provide excellent models for studying the mechanism of susceptible and resistant interactions between plants and pathogens.     

Homolog of FlhDC, a master regulator for flagellum synthesis: required for pathogenicity in Erwinia carotovora subsp. carotovora

 Erwinia carotovora subsp. carotovora (Ecc) is a causal agent of soft-rot diseases in a wide variety of plants. Here, we have isolated nonmotile mutants in Ecc by in vivo insertional mutagenesis using a transposon Tn5. The sequence disrupted by the Tn5 insertion in YMU1 and YMU5 mutants was highly homologous to that of flhC and flhD genes, respectively. They are involved in the initiation of the expression of flagellum-related genes in many gram-negative bacteria such as Escherichia coli and Salmonella. With electron microscopy, the flhC and the flhD homolog mutants were shown to be aflagellate. Furthermore, the virulence of these mutants was greatly reduced in Chinese cabbage and potato compared to that of the parental strain. These results suggest that flagellar formation is required for the pathogenicity of Ecc. Received: November 5, 2002 / Accepted: December 2, 2002 Acknowledgments This research was supported in part by Grant-in-Aid (12052210) and by a grant from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (13073).     

Including an additional systematic environmental effect within a generation in an evaluation model improves accuracy of prediction of breeding values in a closed herd of pigs

The present study evaluated the advantage of mixed‐model techniques over a selection index under different magnitudes of an additional systematic environmental effect (ASEE) in terms of accuracy of prediction and expected genetic gain. The data attempted to simulate a closed herd in a pig breeding program. The base population (G₀) consisted of 10 males and 50 females. Six generations (G₀ to G₅) were selected by using a selection index of three traits without overlapping. Additional systematic environmental constants with four levels in a generation were assigned from a uniform distribution at different ranges. Breeding values of animals in the last generation (G₅) were estimated on the basis of an index of individual phenotype (SI‐U), SI‐U adjusted for ASEE using a least‐squares mean (SI‐A), best linear unbiased prediction using an animal model excluding ASEE (AM‐E), and an animal model including ASEE (AM‐I). Accuracy of prediction and expected genetic gain were larger by the animal model than by the selection index, even if heritability of the traits selected was high and ASEE was set to zero. When ASEE was zero, the accuracy of prediction and expected genetic gain given by SI‐U and AM‐I were similar to those given by SI‐A and AM‐E, respectively. However, the differences in accuracy and expected gain between SI‐U and AI‐A and between AM‐I and AM‐E increased as the range of ASEE increased. It was concluded that selection based on an animal model was more effective than index selection, even if the herd environment was uniform and traits with high heritability were selected, and that it should be always included in an evaluation model, however slight any systematic environmental effect may be in a closed herd.     

Improving management for higher reproduction accelerates genetic improvement in closed herd of swine

The present study compared responses to selection at different conception rates and litter sizes at weaning in a simulated closed herd in a swine breeding program. The base population consisted of 10 males and 50 females, and 10 generations of selection was practiced by using individual phenotype or best linear unbiased prediction of breeding values for a trait with heritability (h²) of either 0.2 or 0.5. The probability of conception in a single mating was assumed to be 0.8, 0.9 or 1.0. Litter size at weaning was sampled randomly from a normal distribution with mean 8, 10 or 12 and variance 8.1225. Genetic response increased by approximately 6% for h² = 0.2 and approximately 5% for h² = 0.5 at generation 10 when conception rate was increased from 0.8 to 1.0. However, litter size at weaning did not affect response to selection. In conclusion, improving conception rate by environmental management increases genetic response indirectly in a breeding program of a closed swine herd.     

Laboratory diagnosis of canine GM2-gangliosidosis using blood and cerebrospinal fluid.

In the present study, laboratory techniques were used to diagnose canine GM2-gangliosidosis using blood and cerebrospinal fluid (CSF) that can be collected noninvasively from living individuals. Lysosomal acid beta-hexosaminidase (Hex) was measured spectrofluorometrically using 4-methylumbelliferyl N-acetyl-beta-D-glucosaminide and 4-methylumbelliferyl 7-(6-sulfo-2-acetamido-2-deoxy-beta-D-glucopyranoside) as substrates. Main isoenzymes A and B of Hex in leukocytes were also analyzed using cellulose acetate membrane electrophoresis. GM2-ganglioside in CSF was detected and determined quantitatively by using thin-layer chromatography/enzyme-immunostaining method with anti-GM2-ganglioside antibody. In normal dogs, Hex activities could be determined in leukocytes, serum, and CSF and the total activities were markedly reduced in all the enzyme sources in a dog with Sandhoff disease. Electrophoresis of a leukocyte lysate from a normal dog showed that the Hex A and Hex B were not separated distinctively with formation of a broad band, whereas there were no bands in electrophoresis of a lysate from a dog with Sandhoff disease, showing a deficiency in the total enzyme activity. GM2-ganglioside could be detected and determined quantitatively in as little as 100 microl of canine CSE GM2-ganglioside in CSF in a dog with Sandhoff disease increased to 46 times the normal level. In conclusion, the methods in the present study are useful for diagnosis of canine GM2-gangliosidosis. These techniques enable definitive and early diagnosis of canine GM2-gangliosidosis even if tissues and organs cannot be obtained.     

Mitochondrial DNA analysis of Nepalese domestic dwarf cattle Lulu*

Dwarf Lulu cattle, the only Bos Taurus type of cattle in Nepal, are raised under severe environments in the mountainous zone of that country. In the present study, the body measurement traits, cytogenetic and molecular genetic characteristics of the Lulu cattle are investigated. Blood samples were collected from 31 animals in four villages (altitudes 2590–3550 m) in the southern part of Mustang. The Lulu cattle had a normal karyotype with 2n = 60, XY or XX. Only one male examined had a large submetacentric X‐chromosome and a small submetacentric taurine type Y‐chromosome. The mitochodrial DNA (mtDNA) genotypes were analyzed by PCR mediated restriction fragment length polymorphisms, displacement (D)‐loop region PCR mediated single strand conformation polymorphisms, and D‐loop region sequences. Many base substitutions were found in the D‐loop region, suggesting that the Lulu cattle originated from at least 10 maternal lines. Three types of mtDNA from these cattle were found, the Bos taurus type (n = 23), the Bos indicus type (n = 6), and the Bos grunniens type (n = 2). In the village at the lowest altitude, four of the five cows were of the Bos indicus type. These results indicated that mtDNA types of the Lulu cattle mostly belong to Bos taurus, but have been hybridized with Bos indicus cattle in lower‐elevation regions in their maternal lineage.     

A survey of antimicrobial resistance in Escherichia coli isolated from wild sika deer (Cervus nippon) in Japan

We examined the antimicrobial susceptibility of 848 Escherichia coli isolates from 237 feces samples of wild sika deer (Cervus nippon) captured between 2016 and 2019 in 39 of the 47 prefectures of Japan. Five of the 237 wild sika deer (2.1%) carried E. coli with resistance to at least one antimicrobial, and all the resistant isolates showed resistance to tetracycline. The resistant isolates contained antimicrobial resistance genes that were similar to those in E. coli derived from humans and farm animals. Although wild sika deer are not currently likely to be a source for the transmission of antimicrobial resistance in Japan, they can potentially mediate antimicrobial resistance spread by coming into contact with humans, animals, and their surroundings.