Fecal ammonia, urea, volatile fatty acid and lactate levels in dairy cows and their pathophysiological significance during diarrhea

Normal fecal samples were taken from lactating cows fed either a total mixed ration (TMR; n = 30) or pasture‐based diet (20) and from dry cows fed mainly on hay (15). Diarrheic fecal samples (n = 51) were collected from 21 sick dairy cows. Fecal analyses of ammonia, urea, lactate and volatile fatty acid (VFA) levels were used to evaluate colonic fermentation. Most normal feces had reasonably neutral pH, however, alkaline feces were observed in diarrheic cows. Although fecal lactate is higher in cows on grazing pasture, lactate levels were generally lower in the cows in the present study. Fecal VFA levels were higher in lactating cows than in dry cows. Elevated fecal urea was observed in diarrheic cows, however, many fecal samples in normal and diarrheic cows contained no urea. Fecal VFA levels in diarrheic cows were lower than in normal lactating cows, but were approximately equivalent to those in dry cows. Grazing or dry cows showed higher acetate and lower n‐butyrate proportions compared with TMR‐fed or diarrheic cows. Higher proportions of branched chain VFAs were observed in diarrheic cows, and the lowest level was observed in grazing cows. The present results indicate that intracolonic nitrogen equilibrium and proteolytic fermentation are altered by diarrheic status.     

Similarity between Copper Resistance Genes from Pseudomonas syringae pv. actinidiae and P. syringae pv. tomato

Twenty-eight strains of Pseudomonas syringae pv. actinidiae isolated in 1984, 1987 and 1988 from kiwifruit orchards in Japan were tested for their resistance to copper sulfate. All strains isolated in 1984 were copper sensitive with a minimum inhibitory concentration (MIC) of cupric sulfate of 0.75 mM. However, some strains isolated in 1987 and 1988 were resistant, with the MIC ranging from 2.25 to 3.0 mM. All copper-resistant strains contained at least one of two plasmids, pPaCul (about 70.5 kb) or pPaCu2 (about 280 kb), or both. In a copper-resistant strain Pa429, the location of the copper-resistance gene(s) was examined by insertional inactivation with Tn5. The MIC of copper sulfate in the copper-sensitive mutant obtained by Tn5 tagging decreased from 2.75 to 0.75 mM. The 14.5 kb BamHI fragment, designated pPaCuB14, containing the same locus mutagenized with Tn5 was cloned from pPaCu1. However, pPaCuB14 did not confer copper resistance in the transformant of copper-sensitive strain Pa21R, suggesting that this clone did not contain a full set of copper-resistance gene(s). Then a cosmid library of pPaCu1 was constructed and six cosmid clones hybridized with pPaCuB14 were selected. One of the six cosmids, designated pPaCuC1, conferred a near wild-type level of copper resistance in the transformant of the copper-sensitive strain. pPaCuC1 had a homologous region that hybridized with all of the PCR-amplifled fragments of copA, copB, copR, and copS genes of P. syringae pv. tomato. DNA sequence analysis of the homologous region revealed the existence of four open reading frames (ORF A, B, R and S) oriented in the same direction. The predicted amino acid sequences of ORF A, B, R and S had 80, 70, 97 and 95% identity with CopA, B, R and S of P. syringae pv. tomato, respectively. Received 5 July 2001/ Accepted in revised form 27 September 2001     

Thermal resistance traits of the clonal silver crucian carp Carassius langsdorfii and evaluation of these traits using primary culture cells

The thermal resistance traits of three clonal lines of the silver crucian carp Carassius langsdorfii (SCC-1, -2 and -3) were investigated. Individual juvenile fish reared at 20°C were exposed to thermal stress at 36.5 ± 0.5°C and their death times during this exposure were measured. The death times of SCC-2 and SCC-3 fish were 66.6 ± 31.2 and 144.7 ± 49.8 min, respectively. In contrast, all SCC-1 fish survived under these conditions. Furthermore, the thermal sensitivity of primary culture cells from each clone was determined at 37, 40 or 43°C by the Trypan blue assay. Under all treatment conditions, the thermal sensitivity of the SCC-1 primary cultures was lower than those of the others. These results estimated the correlation between the in vivo and in vitro thermal resistances. Therefore, the use of primary culture cells to evaluate thermal resistance could be a useful method of selection breeding.     

Impact of bark stripping by sika deer, Cervus nippon, on subalpine coniferous forests in central Japan

We investigated the effect of bark stripping by sika deer, Cervus nippon, on forest regeneration in subalpine coniferous forests on Mt. Ohdaigahara and in the Ohmine Mountains of central Japan. Bark stripping by sika deer occurred in five major tree species: Abies homolepis; Abies Veitchii; Tsuga diversifolia; Picea jezoensis var. hondoensis; and Chamaecyparis obtusa. The percentage of damaged trees on Mt. Ohdaigahara was higher than in the Ohmine Mountains, probably because of the higher deer density. On Mt. Ohdaigahara, the DBH distributions of stems for P. jezoensis var. hondoensis, A. homolepis, T. diversifolia andC. obtusa were bell-shaped with fewer smaller and larger trees. On the other hand, in the Ohmine Mountains the distributions for P. jezoensis var. hondoensis and A. Veitchii showed a reverse-J shaped with more smaller trees. Larger overstory conifers on Mt. Ohdaigahara were killed by bark stripping when 100% barked, although in the Ohmine Mountains ca. 50% of the trees survived even when 100% barked. After the disappearance of the overstory conifers on Mt. Ohdaigahara, the dwarf bamboo, Sasa nipponica, expanded into the forest floor due to changes in light reaching the forest floor. Since S. nipponica is the main forage of deer in this area, this increase caused a corresponding increase in the deer population, which in turn, could cause a further decline in the coniferous forests.     

Relationships between tropomyosin and myosin heavy chain isoforms in bovine skeletal muscle

The composition of tropomyosin (TPM) and myosin heavy chain (MyHC) isoforms was analyzed in 10 physiologically different bovine muscles ( masseter , diaphragm, tongue, semispinalis, pectoralis profundus , biceps femoris, psoas major , semimembranosus, longissimus thoracis and semitendinosus ) to clarify the relationships between TPM and MyHC isoforms in different muscle fiber types. The content of TPM1 and TPM3 was different in muscles according to their function in muscle contraction, although the content of TPM2 was constantly about 50% of the total TPM in all muscles. The content of TPM1 was higher in semimembranosus , longissimus thoracis and semitendinosus, while that of TPM3 was higher in masseter and diaphragm. The high positive correlation between MyHC-slow content and TPM3 content ( r  = 0.92) suggested a coexpression of TPM3 and MyHC-slow isoforms in a muscle fiber. MyHC-slow and TPM3 were expressed at the same level in masseter and diaphragm, whereas there was more TPM3 than MyHC-slow in tongue and semispinalis , so it appears that the excess TPM3 in tongue and semispinalis is expressed with other MyHC isoforms. MyHC-2a was the only fast type isoform expressed in tongue and semispinalis . Therefore, the excess TPM3 was composed of myofibrils with MyHC-2a. The results suggested that a fiber expressing MyHC-2a would be regulated delicately by changing the TPM isoform types.     

Electrotonic junctions between teleost spinal neurons: electrophysiology and ultrastructure

Electrotonic transmission between spinal neurons is correlated with distinctive apposition of cell processes involving membrane fusion. In the same neurons, postsynaptic potentials appear to arise at typical synaptic knobs where there is an intercellular space.     

The effects of concentrations of lysine in media on differentiation of 3T3‐L1 preadipocytes

Intramuscular fat content is increased by feeding of low lysine diets in pigs. Reduction in dietary lysine intake results in low plasma lysine concentration and low cytosolic lysine concentration in skeletal muscles. From these observations, we hypothesized that low plasma lysine concentration in pigs fed on low lysine diets reduced supply of lysine from blood circulation to preadipocytes, and this limited supply of lysine might promote adipocyte differentiation in porcine muscles. In order to verify the hypothesis, we investigated the effects of low concentrations of lysine in culture medium on differentiation of 3T3‐L1 preadipocytes. Low concentration of lysine suppressed lipid accumulation and messenger RNA (mRNA) expression and enzyme activity of fatty acid synthase. mRNA expressions of peroxisome proliferator‐activated receptor γ (PPARγ) and CCAAT/enhancer binding protein α (C/EBPα) were lower in cells cultured in low lysine medium. On the other hand, mRNA and protein expressions of C/EBPβ and C/EBPδ were not inhibited by low concentrations of lysine in culture medium. These results indicate that low lysine concentrations in culture medium inhibit differentiation of 3T3‐L1 preadipocytes through inhibiting the mRNA expressions of PPARγ and C/EBPα.     

Bromocriptine inhibits salsolinol‐induced prolactin release in male goats

The secretion of prolactin (PRL) is under the dominant and tonic inhibitory control of dopamine (DA); however, we have recently found that salsolinol (SAL), an endogenous DA‐derived compound, strongly stimulated the release of PRL in ruminants. The aim of the present study was to clarify the inhibitory effect of DA on the SAL‐induced release of PRL in ruminants. The experiments were performed from late June to early July. Male goats were given a single intravenous (i.v.) injection of SAL (5 mg/kg body weight (BW)), a DA receptor antagonist (sulpiride, 0.1 mg/kg BW), or thyrotropin‐releasing hormone (TRH, 1 µg/kg BW) before and after treatment with a DA receptor agonist (bromocriptine), and the effect of DA on SAL‐induced PRL release was compared to that on sulpiride‐ or TRH‐induced release. Bromocriptine completely inhibited the SAL‐induced release of PRL (P < 0.05), and the area under the response curve (AUC) for a 120‐min period after the treatment with bromocriptine was 1/28 of that for before the treatment (P < 0.05). Bromocriptine also completely inhibited the sulpiride‐induced release (P < 0.05). The AUC post‐treatment was 1/17 that of pre‐treatment with bromocriptine (P < 0.05). Bromocriptine also inhibited the TRH‐induced release (P < 0.05), though not completely. The AUC post‐treatment was 1/3.8 that of pre‐treatment (P < 0.05). These results indicate that DA inhibits the SAL‐induced release of PRL in male goats, and suggest that SAL and DA are involved in regulating the secretion of PRL. They also suggest that in terms of the regulatory process for the secretion of PRL, SAL resembles sulpiride but differs from TRH.