Approach of allelopathy study with Arabidopsis thaliana (L.) Hevnh. and Neurospora crassa

Allelopathy in Arabidopsis seeds was investigated in the present study because there are few available reports of allelopathy studies regarding the early development stage of the plant life cycle, and because Arabidopsis is widely used as a model plant for molecular biology. When cockscomb ( Celosia cristata ) was incubated together with Arabidopsis seeds, the growth of cockscomb was promoted by Arabidopsis seeds. Exudate of Arabidopsis seeds showed a promotive activity on the growth of cockscomb and Arabidopsis itself. A new method for studying allelopathic mechanisms involving the interaction of the plant Arabidopsis thaliana (L.) Hevnh. and the fungus Neurospora crassa was suggested. Arabidopsis seed exudate and the water-soluble fraction of the exudate promoted Arabidopsis growth and reduced fungal germination, indicating that the seed exudate has a species-selective activity. Conversely, the ethylacetate-soluble fraction inhibited growth of all tested materials in the current study. These results demonstrate that the water-soluble substance(s) released from Arabidopsis seeds have species-selective activity on growth of both plants and microorganisms.     

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.     

Single oral β-cryptoxanthin administration increases its serum concentration and enhances peripheral blood neutrophil function in Holstein cattle

We investigated the effect of oral administration of β-cryptoxanthin (β-CRX) on its serum concentration and peripheral neutrophil functions by the chemiluminescence (CL) response in Holstein cattle. A single oral administration of β-CRX was performed for serum β-CRX concentration (0, 0.05, 0.1, or 0.2 mg/kg body weight [BW]) and for peak CL response of peripheral neutrophils (0.2 mg/kg BW). The serum β-CRX concentration was peaked on 2 days after, similar to peak CL response on 3 days after β-CRX administration. Therefore, a single oral administration of β-CRX (0.2 mg/kg BW) induces higher serum concentration and concurrently enhances bactericidal ability of peripheral neutrophils in Holstein cattle.     

Characterization of a novel nicotinamide adenine dinucleotide-cytochrome b5 reductase mutation associated with canine hereditary methemoglobinemia

Hereditary methemoglobinemia associated with nicotinamide adenine dinucleotide-cytochrome b5 reductase (b5R) deficiency is a rare autosomal recessive disorder in animals. Recently, nonsynonymous b5R gene (CYB5R3) variants have been reported to be associated with canine and feline hereditary methemoglobinemia. However, the underlying molecular mechanisms of canine and feline methemoglobinemia caused by these nonsynonymous variants have not yet been reported. Previously, we reported a Pomeranian dog family with hereditary methemoglobinemia, carrying CYB5R3 mutation of an A>C transition at codon 194 in exon 7, replacing an isoleucine residue with leucine (p.Ile194Leu). In this study, we investigated the enzymatic and structural properties of the soluble form of wild-type and Ile194Leu canine b5Rs to characterize the effects of this missense mutation. Our results showed that the kinetic properties of the mutant enzyme were not affected by this amino acid substitution. The secondary structure of the wild-type and Ile194Leu b5Rs detected by circular dichroism showed a similar pattern. However, the mutant enzyme exhibited decreased heat stability and increased susceptibility to trypsin hydrolysis. Moreover, the thermostability and unfolding measurements indicated that the mutant enzyme was more sensitive to temperature-dependent denaturation than the wild-type b5R. We concluded from these results that unstable mutant enzyme properties with normal enzymatic activity would be associated with hereditary methemoglobinemia in the Pomeranian dog family.     

Effects of acid extracts of tomato (Lycopersicon esculentum Mill.) and carrot (Daucus carota L.)wastes from the food industry on the growth of some crops and weeds

The effects of acid extracts of tomato ( Lycopersicon esculentum Mill.) and carrot ( Daucus carota L.) juice wastes from the food industry on the growth of five species of crops and four weeds were examined. The acid extract of the tomato juice waste promoted the shoot and root growth of tomato, Chinese cabbage, corn, and radish, but not the growth of oat seedlings. The extract did not promote the shoot growth of weeds tested, and it inhibited the root growth of the weeds that included barnyardgrass ( Echinochloa oryzicola Ohwi), southern crabgrass ( Digitaria ciliaris Koel.) and Japanese barnyard millet ( Echinochloa utilis Ohwi). The carrot juice waste was also examined as another waste from the food industry. The effect of the acid extract of carrot juice waste showed significant promoting effects on the root growth of Chinese cabbage; however, the effects on other plants were lower than that of tomato juice waste. It also did not inhibit the growth of weeds. These results suggested that the acid extract from tomato juice waste is useful as plant-growth substances because they have a promoting effect on the shoot and root growth of crops, and an inhibitory effect on the root growth of some weeds.     

The effects of potato pulp and feeding level of supplements on digestibility, in situ forage degradation and ruminal fermentation in beef steers

Six ruminally cannulated Wagyu (Japanese Black) steers (average initial bodyweight (BW) 387 ± 29 kg) were used in a split‐plot design experiment, comprising a 3 × 3 Latin square design (whole plot) and a randomized block design (subplot). The whole plot treatments were three different feeding levels of supplemental diet, fed at 0.2, 0.4 and 0.6% of BW, on a dry matter (DM) basis. Subplot treatments were two different supplemental diets: a potato pulp silage‐based diet (PPS) and a grain‐based diet (GRAIN). Chopped, medium‐quality cool‐season grass hay (predominately Timothy, Phleum pratense L) was fed daily at 0.7% BW (on a DM basis) as the basal diet. Each period consisted of 21 d, which included 11 d of adaptation to the diets and 10 d of the collection period. Chromium oxide was used as an indigestible marker. In situ forage degradation was measured using the nylon bag technique. The dry matter intake increased (linear; P < 0.01) as the feeding level increased and was not affected by the diet. Digestibility was not affected by any treatments. The GRAIN diet tended to decrease the rate of in situ forage degradation as the feeding level increased, but this trend was not found in the steers fed the PPS diet. Steers fed the GRAIN diet had a lower (P < 0.1) ruminal pH compared with steers fed the PPS diet. Ruminal pH was not significantly affected by feeding level; however, it was numerically higher for steers supplemented at 0.2% per BW than that for the steers supplemented above 0.4% per BW due probably to the higher starch intake. The total volatile fatty acids concentration numerically increased as the feeding level increased and was not affected by the diet. Increasing the feeding level decreased (linear, P < 0.01) the proportion of acetate. Neither diet nor the feeding level had any effects on the proportion of ruminal propionate. The results suggested that, for steers fed the PPS diet, there are not adverse effects on forage digestion in the rumen that occur as the feeding level is increased.     

Evidence of LAT1 expression in canine caput epididymis

L-type amino acid transporter 1 (LAT1), the first isotype of amino acid transport system L, transports aromatic and branched amino acids pivotal for fundamental cellular activities such cellular growth and proliferation. LAT1 expression was high only in the brain in contrast to its limited distribution and low level of expression in normal tissues. We found potent LAT1 expression in canine caput epididymis by quantitative RT-PCR and Western blotting analysis. Immnuno-histochemical examination revealed observable LAT1 in microvillous epithelial cells.     

Detecting endotoxin activity in bovine serum using an automated testing system

The aim of the present study was to compare the ability of the commercially available portable test system (PTS^TM) to detect endotoxin activity in bovine serum, with that of the traditional LAL-kinetic turbidimetric (KT) and chromogenic (KC) assays. Prior to testing, serum samples, which were obtained from endotoxin-challenged cattle, were diluted 1:20 in endotoxin-free water and heated to 80°C for 10 min. The performance of the PTS^TM was not significantly different from that of the traditional LAL-based assays. The results using PTS^TM correlated with those using KT (r²=0.963, P<0.001) or KC assays (r²=0.982, P<0.001). Based on these findings, the PTS^TM could be applied as a simplified system to assess endotoxin activity in bovine serum.