Molecular detection and characterization of Haemobartonella felis in domestic cats in Japan employing sequence-specific polymerase chain reaction (SS-PCR)

A novel PCR assay was developed in order to examine the prevalence of Haemobartonella felis (H. felis) in Japanese domestic cats and which was able to differentiate of the Ohio strain and the California strain of H. felis. Blood samples from a total of 21 cats suspected of having haemobartonellosis were examined employing a novel PCR assay and demonstrated positive results in 18 cats which was confirmed by cytological examination of blood smears. Four out of 18 positive cats (22%) were infected with the California strain, whilst the other 12 cats (67%) were infected with the Ohio strain and two animals (11%) were infected with both strains. As most of the cats with moderate to severe anemia were infected with the Ohio strain, it is suggested that the most prevalent strain of H. felis in Japanese domestic cats might be the Ohio strain. In the present study, it was thought that molecular detection and characterization of H. felis may provide valuable information regarding the severity and prognosis of this illness.     

Effects of feeding level of milk replacer on body growth,plasma metabolite and insulin concentrations,and visceral organ growth of suckling calves

The objective was to evaluate effects of feeding level of milk replacer on body growth, plasma metabolite and insulin concentrations, and allometric growth of visceral organs in suckling calves. Holstein bull calves (n = 8; 3–4 days of age) were fed either a low amount (average 0.63 kgDM/day, LM) or high amount (average 1.15 kgDM/day, HM) of high protein milk replacer until they were slaughtered at 6 weeks of age. Body weight (BW) at 4, 5, and 6 weeks of age, feed intake, average daily gain, and feed efficiency were higher in the HM than LM calves. The HM group had higher plasma glucose at 3 and 4 weeks of age and insulin levels after the age of 4 weeks compared with LM calves whereas no effect was detected on plasma nonesterified fatty acid or urea nitrogen concentrations. The HM calves had greater empty body weight (EBW), viscera‐free BW and most of the organs dissected than LM calves. Relative weights (% of EBW) of liver, spleen, kidneys, and internal fat were higher, whereas head and large intestine was lower in HM than LM calves. The results suggest that increased milk feeding levels would accelerate the growth of the body and specific organs.     

Moisture Distribution and Texture of Various Types of Cooked Spaghetti

Cooked spaghetti of various types (fresh, dried, frozen, luncheon, and long‐life spaghetti) was examined for moisture distribution by magnetic resonance imaging (MRI). Moisture content was calculated from spinspin relaxation time (T₂) of water proton, based on the correlation between T₂ and moisture content of pulverized durum semolina standard gel samples. Boiled samples of dried and frozen spaghetti had a distinct low moisture region at the center, which was not clearly observed in the other types of boiled spaghetti samples. In particular, the moisture content of boiled long‐life spaghetti was almost homogeneous. Texture of cooked spaghetti samples was evaluated using the force‐distance curve of a mechanical property test. For dried and frozen spaghetti, higher force was observed at the region corresponding to the low moisture core compared with the other types. The luncheon spaghetti and long‐life spaghetti showed a lower breaking force and a larger dip after the peak force representing soft and brittle texture caused by moisture homogeneity. These results indicated that MRI could be used for the quality evaluation of cooked spaghetti through imaging of the moisture distribution, which reflects the mechanical property.     

Bitter gourd suppresses lipopolysaccharide-induced inflammatory responses

Bitter gourd ( Momordica charantia L.) is a popular tropical vegetable in Asian countries. Previously it was shown that bitter gourd placenta extract suppressed lipopolysaccharide (LPS)-induced TNFalpha production in RAW 264.7 macrophage-like cells. Here it is shown that the butanol-soluble fraction of bitter gourd placenta extract strongly suppresses LPS-induced TNFalpha production in RAW 264.7 cells. Gene expression analysis using a fibrous DNA microarray showed that the bitter gourd butanol fraction suppressed expression of various LPS-induced inflammatory genes, such as those for TNF, IL1alpha, IL1beta, G1p2, and Ccl5. The butanol fraction significantly suppressed NFkappaB DNA binding activity and phosphorylation of p38, JNK, and ERK MAPKs. Components in the active fraction from bitter gourd were identified as 1-alpha-linolenoyl-lysophosphatidylcholine (LPC), 2-alpha-linolenoyl-LPC, 1-lynoleoyl-LPC, and 2-linoleoyl-LPC. Purified 1-alpha-linolenoyl-LPC and 1-linoleoyl-LPC suppressed the LPS-induced TNFalpha production of RAW 264.7 cells at a concentration of 10 microg/mL.     

Alpha-eleostearic acid and its dihydroxy derivative are major apoptosis-inducing components of bitter gourd

Bitter gourd ( Momordica charantia L.) pericarp, placenta, and seed extracts were previously shown to induce apoptosis in HL60 human leukemia cells. To determine the active component that induces apoptosis in cancer cells, bitter gourd ethanol extract was fractionated by liquid-liquid partition and silica gel column chromatography. Several fractions obtained by silica gel column chromatography inhibited growth and induced apoptosis in HL60 cells. Among them, fraction 7 had the strongest activity in inhibiting growth and inducing apoptosis in HL60 cells. A component that induced apoptosis in HL60 cells was then isolated from fraction 7 by another silica gel column chromatography and high-performance liquid chromatography (HPLC) using a C18 column and was identified as (9Z,11E,13E)-15,16-dihydroxy-9,11,13-octadecatrienoic acid (15,16-dihydroxy alpha-eleostearic acid). 15,16-Dihydroxy alpha-eleostearic acid induced apoptosis in HL60 cells within 5 h at a concentration of 160 microM (50 microg/mL). (9Z,11E,13E)-9,11,13-Octadecatrienoic acid (alpha-eleostearic acid) is known to be the major conjugated linolenic acid in bitter gourd seeds. Therefore, the effect of alpha-eleostearic acid on the growth of some cancer and normal cell lines was examined. alpha-Eleostearic acid strongly inhibited the growth of some cancer and fibroblast cell lines, including those of HL60 leukemia and HT29 colon carcinoma. alpha-Eleostearic acid induced apoptosis in HL60 cells after a 24 h incubation at a concentration of 5 microM. Thus, alpha-eleostearic acid and the dihydroxy derivative from bitter gourd were suggested to be the major inducers of apoptosis in HL60 cells.     

Changes of plasma free amino acid concentrations and myofibrillar proteolysis index by starvation in non-pregnant dry cows

The effects of 5 day starvation on urinary metabolite excretion, plasma metabolites and free amino acid concentrations in four non‐pregnant dry cows were investigated. Starvation significantly reduced (P < 0.05) the bodyweight and feces weight. The plasma glucose concentration decreased (P < 0.05) during days 2–5 of starvation. Starvation significantly increased (P < 0.05) the plasma nonesterified fatty acid concentration. The plasma urea nitrogen concentration and urinary urea nitrogen excretion increased transiently (P < 0.05) at days 1 and 2 of starvation. The plasma 3‐methlhistidine concentration was significantly higher (P < 0.05) at days 2, 3 and 5 of starvation. However, the urinary 3‐methylhistidine excretion was significantly higher (P < 0.05) at days 4 and 5 of starvation, not corresponding to the change in the plasma. Starvation did not change the plasma proline, methionine, lysine and total amino acid concentrations. On the other hand, starvation reduced (P < 0.05) the plasma aspartic acid, serine, asparagine, glutamic acid, glutamine, alanine, tyrosine, tryptophan and histidine concentrations. Plasma glycine increased during starvation. Plasma threonine, isoleucine, leucine and branched‐chain amino acid increased (P < 0.05) by prolonged starvation. The results of this study indicate that prolonged starvation accelerated myofibril protein degradation in non‐pregnant dry cows and apparently consumes most glucogenic amino acids in preference to other amino acids.     

IL‐12p40 gene expression in lung and hilar lymph nodes of MPS‐resistant pigs

Mycoplasma pneumonia of swine (MPS) is caused by Mycoplasma hyopneumoniae (M.hp) and is a common chronic respiratory disease of pigs. Recently, a genetically selected variant of the Landrace pig (Miyagino L2) has a lower incidence of pulmonary MPS lesions. We investigated the pathological and immunological characteristics of MPS resistance in these pigs (n = 24) by comparing with the normal landrace pig (control: n = 24). The pathological MPS lung lesion score in MPS‐selected landrace pigs was significantly lower than in the control. The gene expression of interleukin (IL)‐12p40, which acts as a chemoattractant and a component of the bioactive cytokines IL‐12 and IL‐23, was significantly higher at the hilar lymph nodes, lung, and spleen in MPS‐selected landrace pigs than in control landrace pigs, and these were negatively correlated with the macroscopic MPS lung lesion score. In summary, we demonstrate that resistance against MPS in Miyagino L2 pigs is associated with IL‐12p40 up‐regulation, in comparison with normal landrace pigs without the MPS vaccine. In addition, a comparative study of macroscopic MPS lung lesions and IL‐12p40 gene expression in lung and hilar lymph nodes may lead to beneficial selection traits for the genetic selection for MPS resistance in pigs.     

Effects of elevated atmospheric CO2 concentration on the nutrient uptake characteristics of Japanese larch (Larix kaempferi)

We evaluated the response of Japanese larch (Larix kaempferi Sieb. & Zucc.) to elevated atmospheric CO(2) concentration ([CO(2)]) (689 +/- 75 ppm in 2002 and 697 +/- 90 ppm in 2003) over 2 years in a field experiment with open-top chambers. Root activity was assessed as nitrogen, phosphorus and potassium uptake rates estimated from successive measurements of absorbed amounts. Dry matter production of whole plants was unaffected by elevated [CO(2)] in the first year of treatment, but increased significantly in response to elevated [CO(2)] in the second year. In contrast, elevated [CO(2)] increased the root to shoot ratio and fine root dry mass in the first year, but not in the second year. Elevated [CO(2)] had no effect on tissue N, P and K concentrations. Uptake rates of N, P and K correlated with whole-plant relative growth rates, but were unaffected by growth [CO(2)], as was ectomycorrhizal colonization, a factor assumed to be important for nutrient uptake in trees. We conclude that improved growth of Larix kaempferi in response to elevated [CO(2)] is accompanied by increased root biomass, but not by increased root activity.     

Association between bacterial community structures and mortality of fish larvae in intensive rearing systems

Bacterial community structures were analyzed in water used for rearing fish larvae by fluorescence in situ hybridization. In Experiment 1, red sea bream Pagrus major larvae were reared in two commercial seed production tanks. The survival rate in Tank 1 was higher than in Tank 2, even though phytoplankton, Nannochloropsis sp., was added to both tanks. In Tank 2, γ-proteobacteria became dominant (∼70% of total bacteria) on day 13, there after heavy larval mortalities occurred. In Tank 1, however, α-proteobacteria and the Cytophaga-Flavobacterium cluster were predominant from day − 1 until day 13; no significant mortality was recorded. In Experiment 2, marble goby Oxyeleotris marmoratus larvae were cultured with or without Nannochloropsis sp. At the end of the experiment, larval survival rates in aquaria with Nannochloropsis sp. were significantly (P <0.05) higher than those without. In rearing water without Nannochloropsis sp., γ-proteobacteria increased during rearing. In rearing water with Nannochloropsis sp., α-prote obacteria and the Cytophaga-Flavobacterium cluster were predominant at the beginning of the experiments and the relative abundance of γ-proteobacteria was maintained at a lower level throughout the experiments. The predominance of α-proteobacteria and the Cytophaga-Flavobacterium cluster appears to be a good indicator of successful larval production.