Classification of lactic acid bacteria isolated from chigee and mare milk collected in Inner Mongolia

One hundred and seventeen isolates from chigee and 191 isolates from mare milk collected in Inner Mongolia were classified into six and 10 groups, respectively, based on the protein profiles obtained by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE). A dendrogram on the basis of the molecular sizes of the protein bands was drawn and the similarity among the groups ranged from 96% to 44%. Representative isolates from all groups were identified by 16S rDNA sequencing and a phylogenetic tree was drawn. Furthermore, the results of carbohydrate fermentation tests of all groups were compared with those obtained by 16S rDNA sequencing and SDS‐PAGE analyzes. The isolates from chigee consisted of mainly Lactobacillus sp., Lactobacillus (Lb.) plantarum, Lb. pentosus and Lactococcus (Lc.) lactis ssp. cremoris were isolated at the rates of 48, 33 and 19%, respectively. The isolates from mare milk consisted of coccus, namely, Leuconostoc (Leuc.) mesenteroides, Leuc. pseudomesenteroides, Lc. garvieae, Lc. lactis ssp. lactis, Streptococcus (Sc.) parauberis and Enterococcus (Ec.) faecium. The isolation rates were 45, 19, 7, 8, 16 and 6%, respectively. The results demonstrated that the species isolated from chigee were different from those in mare milk.     

High prevalence of Mycoplasma equirhinis in Thoroughbred horses with respiratory symptoms in autumn 2018

Mycoplasma species are often isolated from horses with respiratory symptoms; however, the pathogenicity of Mycoplasma is still unclear. In autumn of 2018, we encountered an increase in cases with respiratory symptoms, mainly coughing, in a group of Thoroughbred racehorses in Japan. We examined tracheal wash samples obtained from 40 of those cases. Bacteria and viruses that commonly cause respiratory symptoms were investigated, and anaerobes were detected in only 5 cases and Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) was detected in only 1 case of 40 cases with loop-mediated isothermal amplification assay. S. zooepidemicus and Streptococcus pneumoniae were isolated at a bacterial count of higher than 1.0 × 10⁴ CFU/ml from 5 and 2 cases of 28 cases cultured, respectively. None of the viruses investigated was detected in 40 cases. Mycoplasma equirhinis (M. equirhinis) was isolated from 40.0% (16/40) of the cases, which was higher than previously reported isolation rates. The rate of M. equirhinis isolation in the cases from 2018 was significantly higher than the isolation rates in the other horses: clinical cases with respiratory symptoms in 2019–2020 (13.6%, 3/22) and healthy horses (13.5%, 5/37) in Japan. In this study, the isolation rate of M. equirhinis from horse group with cough symptoms in 2018 was high and no other common etiological agents were detected. The pathogenesis of M. equirhinis is still unclear, however, M. equirhinis might have been associated with respiratory symptoms in the Thoroughbred horse cases in 2018.     

A review of tower flux observation sites in Asia

Aggregating and sharing the metadata of flux observation sites results in a strong collaboration among various fields of study. Such data sharing will also be a part of the future design of a tower flux observation network in Asia. The aim of this review is to comprehend the state of tower flux observation sites in Asia. There are 109 tower flux observation sites in Asia including 51 forest sites. There are more new sites under construction in Asia than in America and Europe. These sites range from the taiga in Siberia to the rainforest in Southeast Asia, and from the equatorial to polar Koeppen climate zones. There are many highly humid areas in Asia, not only at low latitudes but also at middle latitudes. This climate condition has developed unique vegetation such as lucidophyllous (evergreen broadleaf) forest, which is distributed in warm areas with high precipitation in the growing season. However, there are only a few observations taking place in lucidophyllous forest. Rice paddy fields are also unique land cover in Asia. It is important to accumulate long-term data for rice fields with their management records, because plant activity depends highly on both climate conditions and land-use management. Flux data, especially net ecosystem exchange and related elements, are used for widespread studies not only within the flux-research community but also in other fields of study, for example remote sensing. At present, however, both the quantity and quality of the data are not sufficient for these studies. Regarding the quantity, there are many recently established sites that have not published data yet; regarding quality, flux data include uncertainties caused by methodological problems. Flux researchers are required not only to obtain flux data but also to improve their quality. Meanwhile, data users must understand there are still uncertainties in flux data. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Estimating a suitable microsatellite marker set for individual identification and parentage tests of brown bear (Ursus arctos) in the Akan–Shiranuka Region, eastern Hokkaido, Japan

We examined 24 microsatellite markers to select those most suitable for individual identification and parentage tests of brown bears (Ursus arctos) based on statistical parameters and experimental error using liver samples (38 issues) from nuisance bears killed in the Akan–Shiranuka Region, eastern Hokkaido, Japan from 1996 to 2006. We found seven microsatellite markers suitable for identifying individuals (G1A, G10B, G10L, UarMU5, UarMU23, UarMU50, and UarMU51; probability of identity P _id = 3.17  × 10⁻⁷; probability of identity of sibs P _id-sib  = 2.23 × 10⁻³), 12 microsatellite markers suitable for parentage testing when one parent is known [G1A, G1D, G10B, G10L, G10P, UarMU5, UarMU23, UarMU50, UarMU51, UarMU59, UarMU61, and UarMU64; probability of excluding one parent (PE I = 0.9991)], and 15 microsatellite markers for parentage testing when neither parents are known [three markers added to the above: G10M, G10X, and UarMU9; probability of excluding both parents (PE II = 0.9869)].     

Automatic counting of FISH-labeled microbes by an LED illuminated detecting apparatus

ABSTRACT:   The Bioplorer (BP) (Matsushita Ecology Systems Co. Ltd, Kasugai, Aichi-ken, Japan) is an apparatus that consists of a light-emitting diode and optical and image analysis systems. This instrument has been developed and used for the semi-automatic counting of total microbes, mainly in food, cosmetics and industrial products. In this study, the applicability of BP to the detection and enumeration of eukaryotic and prokaryotic cells, after the fluorescent in situ hybridization (FISH) procedure, was examined. The cells of a yeast ( Candida albicans ) and a bacterium ( Escherichia coli ) were specifically labeled using an oligonucleotide probe with fluorochrome and then counterstained with 4', 6-diamidino-2-phenylindole (DAPI). The numbers of cells after FISH treatment and DAPI staining agreed well, indicating that the BP optical device and image analysis system had enough sensitivity to detect and quantify eukaryotic and prokaryotic cells. Because of its simplicity and reliability, BP can be a new tool for quantification of specifically labeled target microorganisms, both in industrial products and natural samples.     

Spatial distribution of the Japanese common squid, Todarodes pacificus, during its northward migration in the western North Pacific Ocean

The spatial distribution of Todarodes pacificus in and near the Kuroshio/Oyashio Transition Zone during its northward migration was examined by comparative surveys using two types of mid‐water trawl net and supplementary squid jigging from June to July 2000. The vertical and horizontal distribution patterns varied for different body sizes in relation to the oceanographic structure. Todarodes pacificus of 1–20 cm dorsal mantle length (ML) were widely distributed from the coastal waters of Japan to near 162°E longitude, probably due to transport by the Kuroshio Extension (KE). Todarodes pacificus smaller than 10 cm ML were mainly distributed in temperate surface layers at sea surface temperatures (SSTs) >15°C near the KE meander probably because of their poor tolerance to lower temperatures and limited swimming ability. Squid of 10–15 cm ML were distributed in the offshore waters of 10–15°C SST and in the coastal waters of northern Honshu, and underwent diel vertical migrations between the sea surface at night and deeper layers during the daytime. Squid larger than 15 cm ML were distributed in the coastal feeding grounds of northern Honshu and Hokkaido until they began their southward spawning migration. They also underwent diel vertical migrations, but remained deeper at night than the squid of 10–15 cm ML; this migration pattern closely matched that of their main prey such as euphausiids. We concluded that as T. pacificus grow, they shift their distribution range from the temperate surface layer around the KE toward the colder deeper layers, above 5°C, in the Oyashio and coastal areas.     

A possible function of Nik-related kinase in the labyrinth layer of delayed delivery mouse placentas

In mice and humans, Nik-related protein kinase (Nrk) is an X-linked gene that encodes a serine/threonine kinase belonging to GCK group 4. Nrk knockout (Nrk KO) mice exhibit delayed delivery, possibly due to defective communication between the Nrk KO conceptus and its mother. However, the mechanism of delayed labor remains largely unknown. Here, we found that in pregnant mothers with the Nrk KO conceptus, the serum progesterone (P4) and placental lactogen (PL-2) concentrations in late pregnancy were higher than those in the wild type. Moreover, we demonstrated that Nrk is expressed in trophoblast giant cells (TGCs) and syncytiotrophoblast-2 (SynT-2) in the labyrinth layer of the mouse placenta. In the human placenta, NRK is also expressed in Syn-T in villi. Both human Syn-T and mouse TGCs of the labyrinth layer are present within fetal tissues that are in direct contact with the maternal blood. The labyrinth layer of the Nrk KO conceptus was gigantic, with enlarged cytoplasm and Golgi bodies in the TGCs. To investigate the function of Nrk in the labyrinth layer, a differentially expressed gene (DEG) analysis was performed. The DEG analysis revealed that labor-promoting factors, such as prostaglandins, were decreased, and pregnancy-maintaining factors, such as the prolactin family and P4 receptor, were increased. These findings suggest that the Nrk KO mice exhibit delayed delivery owing to high P4 concentrations caused by the hypersecretion of pregnancy-maintaining factors, such as PL-2, from the placenta.     

Spontaneous Gingivitis Related to Hair Penetration in Rats

Maxillary gingivae from male and female Crl:CD(SD) rats at 12, 16, 21, and 34 weeks of age were examined histologically. The incidence of gingivitis was approximately 40%, with no age or sex predilection, and was most frequent between the first and second molar. Lesions were characterized by acute focal neutrophilic infiltration into the gingival mucosa, occasionally with inflammatory exudate. In severe cases, inflammation extended to the periodontal ligament with abscess formation, and adjacent alveolar bone destruction/resorption. The most characteristic finding was the presence of hair shafts associated with the lesion, which was observed in approximately 80% of the rats with gingivitis. These findings suggest that molar gingivitis occurs in rats from an early age and persists thereafter, and that the main cause of gingivitis in rats is hair penetration into the gingiva. It would be prudent to keep these background lesions in mind as potential modifiers in toxicity studies.     

Acute Phase Pulmonary Responses to a Single Intratracheal Spray Instillation of Magnetite (Fe3O4) Nanoparticles in Fischer 344 Rats

Iron nanomaterials are of considerable interest for application to nanotechnology-related fields including environmental catalysis, biomedical imaging, drug delivery and hyperthermia, because of their superparamagnetic characteristics and high catalytic abilities. However, information about potential risks of iron nanomaterials is limited. The present study assessed pulmonary responses to a single intratracheal spray instillation of triiron tetraoxide nanoparticles (magnetite) in rats. Ten-week-old male and female Fischer 344 rats (n=5/group) were exposed to a single intratracheal spray instillation of 0 (vehicle), 5.0, 15.0 or 45.0 mg/kg body weight (BW) of magnetite. After 14 days, the rats were sacrificed, and biological consequences were investigated. The lung weights of the 15.0 and 45.0 mg/kg BW male and female groups were significantly higher than those of the control groups. The lungs of treated rats showed enlargement and black patches originating from the color of magnetite. The typical histopathological changes in the lungs of the treated rats included infiltration of macrophages phagocytosing magnetite, inflammatory cell infiltration, granuloma formation and an increase of goblet cells in the bronchial epithelium. The results clearly show that instilled magnetite causes foreign body inflammatory and granulating lesions in the lung. These pulmonary responses occur in a dose-dependent manner in association with the increase in lung weight.