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.     

Relationship between anti-insulin antibody production and severe insulin resistance in a diabetic cat

A 5-year-old castrated male domestic shorthair cat was diagnosed with diabetic ketoacidosis and severe insulin resistance. Although the conventional treatment for diabetic ketoacidosis was provided, the cat required frequent hospitalization because of severe dehydration and repeated diabetic ketoacidosis. We detected anti-insulin antibodies for human in this cat. Serum insulin-binding IgG levels were markedly elevated compared with those in healthy cats and other diabetic cats. We initiated prednisolone to suppress the effects of anti-insulin antibodies. After initiation of prednisolone, the cat was gradually recovered with increasing activity and appetite. Furthermore, satisfactory glycemic control was achieved with combined subcutaneous injection of insulin detemir and insulin degludec.     

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.     

Comparative functional anatomy of hindlimb muscles and bones with reference to aquatic adaptation of the sea otter

Although the sea otter (Enhydra lutris) is a complete aquatic species, spending its entire life in the ocean, it has been considered morphologically to be a semi-aquatic animal. This study aimed to clarify the unique hindlimb morphology and functional adaptations of E. lutris in comparison to other Mustelidae species. We compared muscle mass and bone measurements of five Mustelidae species: the sea otter, Eurasian river otter (Lutra lutra), American mink (Neovison vison), Japanese weasel (Mustela itatsi) and Siberian weasel (M. sibirica). In comparison with the other 4 species, E. lutris possessed significantly larger gluteus, popliteus and peroneus muscles, but smaller adductor and ischiopubic muscles. The popliteus muscle may act as a medial rotator of the crus, and the peroneus muscle may act as an abductor of the fifth toe and/or the pronator of the foot. The bundles of the gluteus superficialis muscle of E. lutris were fused with those of the tensor fasciae latae muscle and gluteofemoralis muscles, and they may play a role in femur abduction. These results suggest that E. lutris uses the abducted femur, medially rotated crus, eversion of the ankle and abducted fifth digit or extended interdigital web as a powerful propulsion generator. Therefore, we conclude that E. lutris is a complete aquatic animal, possessing differences in the proportions of the hindlimb muscles compared with those in other semi-aquatic and terrestrial mustelids.     

Genetic variants of the unsaturated fatty acid receptor GPR120 relating to obesity in dogs

G protein-coupled receptor (GPR) 120 is an unsaturated fatty acid receptor, which is associated with various physiological functions. It is reported that the genetic variant of GPR120, p.Arg270His, is detected more in obese people, and this genetic variation functionally relates to obesity in humans. Obesity is a common nutritional disorder also in dogs, but the genetic factors have not ever been identified in dogs. In this study, we investigated the molecular structure of canine GPR120 and searched for candidate genetic variants which may relate to obesity in dogs. Canine GPR120 was highly homologous to those of other species, and seven transmembrane domains and two N-glycosylation sites were conserved. GPR120 mRNA was expressed in lung, jejunum, ileum, colon, hypothalamus, hippocampus, spinal cord, bone marrow, dermis and white adipose tissues in dogs, as those in mice and humans. Genetic variants of GPR120 were explored in client-owned 141 dogs, resulting in that 5 synonymous and 4 non-synonymous variants were found. The variant c.595C>A (p.Pro199Thr) was found in 40 dogs, and the gene frequency was significantly higher in dogs with higher body condition scores, i.e. 0.320 in BCS4–5 dogs, 0.175 in BCS3 dogs and 0.000 in BCS2 dogs. We conclude that c.595C>A (p.Pro199Thr) is a candidate variant relating to obesity, which may be helpful for nutritional management of dogs.     

Cytotoxicity of surfactants to the FHM-sp cell line

Cytotoxicity of eight surfactants was determined by the neutral red assay with the fathead minnow (FHM)-sp cell line, a cell line in suspension culture from fish. The toxicity ranking of the surfactants was benzalkonium chloride > benzethonium chloride > sodium linear - dodecylbenzene–sulfonate (LAS) > potassium laurate > sodium dodecylsulfate > polyoxyethylene(20)sorbitan monolaurate > polyoxyethylene(20)sorbitan monooleate > betaine. The toxicity ranking of the surfactants classified into four groups based on the ion of the hydrophilic group was cationic surfactants > anionic surfactants > non-ionic surfactants > amphipathic surfactants. The FHM-sp cells, as well as the chinook salmon embryo (CHSE)-sp cells, could be inoculated directly to the microplate wells without dispersion by trypsin treatment of cell sheets at room temperature. Therefore, the cytotoxicity assay of the surfactants could be carried out quickly by using the FHM-sp cell line. The FHM-sp cell line had similar or higher sensitivity to sodium dodecylsulfate compared with several cell lines from mammals. The cytotoxicity assay could be shortened by the procedure exposing the surfactants to the FHM-sp cells before the cell monolayer formation in the microplate wells. To use the FHM-sp cell line as a screening tool prior to in vivo testing, studies on the correlation between in vivo data and in vitro data on the toxicity of surfactants are necessary.     

Inheritance mode of microsatellite DNA markers and their use for kinship estimation in kuruma prawn Penaeus japonicus

The allelic inheritance mode of microsatellite DNA markers was examined using seven copulated wild females and their offspring. Five microsatellite loci, CSPJ002 *, CSPJ010 *, CSPJ012 *, CSPJ014 *, and CSPJ015 *, were used in the study. At almost all family/locus combinations, one sire was determined and distributions of genotypes in offspring were consistent with the Mendelian segregation ratio. Distributions of genotypes were consistent with the ratio after assuming a null allele at some loci. Consequently, the alleles of CSPJ002 * and CSPJ012 * were inherited following the Mendelian inheritance mode in every family; however, the null allele was expected in CSPJ010 *, CSPJ014 *, and CSPJ015 * in some families. Thus, these loci should be used carefully in population genetic analysis, but siblings could be detected in the dendrograms based on unweighted pair-group method using arithmetic averages (UPMGA).