Cloned cows with short telomeres deliver healthy offspring with normal-length telomeres

Dolly, the first mammal cloned from a somatic cell, had shorter telomeres than age-matched controls and died at an early age because of disease. To investigate longevity and lifetime performance in cloned animals, we produced cloned cows with short telomeres using oviductal epithelial cells as donor cells. At 5 years of age, despite the presence of short telomeres, all cloned cows delivered multiple healthy offspring following artificial insemination with conventionally processed spermatozoa from noncloned bulls, and their milk production was comparable to that of donor cows. Moreover, this study revealed that the offspring had normal-length telomeres in their leukocytes and major organs. Thus, cloned animals have normal functional germ lines, and therefore germ line function can completely restore telomere lengths in clone gametes by telomerase activity, resulting in healthy offspring with normal-length telomeres.     

Molecular mapping of quantitative trait loci for domestication traits and β-glucan content in a wheat recombinant inbred line population

Genetic maps are useful for analysis of quantitative trait loci (QTLs) and for marker-assisted selection (MAS) in breeding. A simple sequence repeat (SSR) marker linkage map of common wheat was constructed based on recombination inbred lines (RILs) derived from a cross between Chinese Spring and spelt wheat. The map included 264 loci on all wheat chromosomes covering 2,345.2 cM with 962, 794.6, and 588.6 cM for the A, B, and D genomes, respectively. Using the RILs and the map, we detected 42 putative QTLs on 15 chromosomes for ear length, spikelet number, spike compactness, kernel length, kernel width, kernel height and β-glucan content. Each QTL explained 4–45% of the phenotypic variation. Five QTL cluster regions were detected on chromosomes 1A, 5AL, 2B, 2D, and 4D. The first QTLs for β-glucan content in wheat were identified on chromosomes 3A, 1B, 5B, and 6D.     

Soil respiration and methane flux in adjacent forest,grassland, and cornfield soils in Hokkaido,Japan

Soil respiration and methane flux from adjacent forest, grassland, and cornfield were measured by using the closed chamber method from June to November, 1999 in Shizunai, Hokkaido, Japan, where the soil was an Aquic Humic Udivitrands derived from volcanic ash. The forest soil absorbed methane, at arate ranging from -0.12 to -0.02 mg C m_-2 h_-1, while the grassland soil emitted methane, at a rate ranging from undetectable levels to 0.18 mg C m_-2 h_-1. In the cornfield soil methane flux ranged from -0.01 to 0.04 mg C m_-2 h_-1. The soil respiration rate varied from 3 to 230 mg C m_-2 h_-1, 27 to 372 mg C m_-2 h_-1, and 29 to 156 mg C m_-2 h_-1 for the cornfield, grassland, and forest soils, respectively. Linear regression analysis demonstrated that the methane flux rate was positively correlated with the soil water-filled pore space (WFPS), and negatively correlated with the relative gas diffusion coefficient (D/D _o) and air-filled pore space (AFPS). Soil respiration rates were positively correlated with the soil temperature at all the sites. The Q ₁₀ value was 4.8, 3.3, and 1.9 for the cornfield, grassland, and forest soils, respectively.     

Malignant transformation of T-cell large granular lymphocyte leukemia in a dog

An 8-year-old spayed female Golden Retriever was referred to us for evaluation of mild lymphocytosis. The peripheral lymphocytes were comprised of mostly large granular lymphocytes (LGLs), and flow cytometry showed that they were mostly CD3+8+ T lymphocytes. Clonal rearrangement of the T-cell receptor gene was identified in the peripheral blood, and the dog was therefore diagnosed with LGL chronic leukemia. The dog was subclinical without treatment until hospitalization on day 154, at which point the lymphocytes looked like lymphoblasts and the surface markers changed to CD3-8-. This was regarded as malignant transformation from LGL chronic leukemia to the acute type. Sequential chemotherapy was started, but the dog died on day 190. Necropsy revealed tumor cell infiltration into the heart, skin, and brain.     

Growing porcine oocyte-granulosa cell complexes acquired meiotic competence during in vitro culture

The aim of this study was to establish a culture system to improve the meiotic competence of porcine oocyte-granulosa cell complexes (OGCs) obtained from preantral or early antral follicles. Porcine OGCs were recovered from follicles with diameters of 230-300 (preantral follicles), 300-500, and 500-700 mum (early antral follicles) using scalpels. The OGCs were cultured for 2 weeks in culture medium. We examined the effects of the sizes of the follicles from which OGCs were recovered, the concentrations of polyvinylpyrrolidone (PVP, 0-8%) in the culture medium, and 2 types of culture dish (Falcon 3002 vs 1007) on formation of the antrum of OGCs. After culture, the oocytes were matured for 44 h to assess their meiotic competence. OGCs recovered from small follicles (230-500 microm) required longer (P<0.05) than larger follicles to form the antrum structure. The percentage of OGCs forming the antrum structure that were cultured in 2% PVP (31%) was higher (P<0.05) than for those cultured in other PVP concentrations (0-11%). The percentages of antrum-structure formation for OGCs cultured on Falcon 3002 (83% for 2% PVP and 60% for 4% PVP) were higher (P<0.05) than those cultured on Falcon 1007 (47% for 2% PVP and 9% for 4% PVP). Furthermore, all of the intact oocytes that were obtained from culture of OGCs and that formed an antrum were in the GV stage (n=28). When these immature oocytes were cultured for 44 h, the percentage of oocytes that reached the metaphase II stage (25%, n=68) was higher (P<0.0001) than that of oocytes matured without culture (0.7%, n=137). The results of the present study show that porcine OGCs obtained from preantral or early antral follicles acquire meiotic competence in vitro.     

Changes of multi-drug resistance pattern in Salmonella enterica subspecies enterica serovar typhimurium isolates from food-producing animals in Japan

A total of 153 isolates of Salmonella Typhimurium derived from food-producing animals in Japan between 2002 and 2005 were investigated for antimicrobial resistance and phage types related to definitive phage type 104 (DT104). The predominant resistance type was resistance to ampicillin, dihydrostreptomycin, kanamycin, and oxytetracycline in bovine (45.2%, 48/104) and resistance to dihydrostreptomycin and oxytetracycline in porcine isolates (58.7%, 27/48). DT104-related phage type was found in 32 of 104 bovine isolates, two of 48 porcine isolates, and one of eight isolates from poultry, showing that the proportion of the phage type in S. Typhimurium isolates from cattle and pigs significantly (P<0.01) decreased from 71.9% and 31.4% in 1999-2001 to 30.8% and 4.1% in 2002-2005, respectively.     

Antimicrobial susceptibility of Mannheimia haemolytica isolates from cattle in Japan from 2001 to 2002

A total of 27 clinical isolates of Mannheimia haemolytica from cattle in Japan from 2001 to 2002 were examined for antimicrobial susceptibility to 25 antimicrobial agents. The minimum inhibitory concentrations of 25 different antimicrobials were determined by an agar dilution method according to the guidelines of the National Committee for Clinical Laboratory Standards. Of the 27 isolates, seven isolates (26.9%) were resistant to at least one of the 25 drugs and resistance rates ranged from 3.7 to 18.5%. Resistance rates to dihydrostreptomycin (18.5%), oxytetracycline (11.1%), and doxycycline (11.1%) were relatively high and those to the remaining drugs were less than 10%.     

In vitro activity of 24 antimicrobial agents against Staphylococcus and Streptococcus isolated from diseased animals in Japan

A total of 88 Staphylococcus and 61 Streptococcus isolates from diseased animals throughout Japan were examined in 2000 for the minimum inhibitory concentrations of 24 different antimicrobials by the agar dilution method standardized by the Japanese Society of Chemotherapy. The resistance rates to aminobenzylpenicillin (36.4%) and benzylpenicillin (35.2%) were high in Staphylococcus isolates, and those to oxytetracycline (45.9%) and kanamycin (21.3%) were high in Streptococcus isolates. Two isolates resistant to oxacillin harbored the mecA gene. One was Staphylococcus epidermidis derived from a pig with arthritis, and the other Staphylococcus cohnii from a head of cattle with mastitis.