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.     

Type VII and XVII Collagen mRNA Expressions in Regenerated Epidermal Laminae in Chronic Equine Laminitis

To confirm ability forming the basement membrane of the regenerated laminar epidermis (rLE) in chronic laminitis, expression of type VII and type XVII collagen mRNAs in the rLE was studied applying sequences of two type of murine collagens. On northern blot analysis, complement DNA (cDNA) probes adjusted from the murine type VII and type XVII collagen could hybridize with the equine mRNAs, and each signal was detected as single-bands at approximately 9.5 kb and 5.6 kb, respectively. Contrasting with the expression level of equine glyceraldehyde-3-phosphate dehydrogenease mRNA, the band of type VII collagen mRNA in laminitis was stronger than normal, but the type XVII collagen mRNA in laminitis was less than normal. By in situ hybridization, positive signals in response to the murine type VII and type XVII collagen mRNA probes could be detected in the equine laminitic rLE region. From these results, it is concluded that the keratinocytes constructing the rLE in chronic stage of laminitis can express type VII and type XVII collagen mRNAs and these expression patterns were different from the normal.     

Immunohistochemical study on the delayed progression of epithelial apoptosis in follicle-associated epithelium of rat Peyer's patch

It is well known that some caspases in apoptosis is involved in determinant of terminal differentiation and maturation of various cells. Our previous study ultrastructurally clarified the differentiation into M cells from immature microvillous epithelial cells and the redifferentiation from M cells to microvillous epithelial cells in the follicle-associated epithelium (FAE) of rat Peyer's patch. In this study, the difference of epithelial apoptosis between the FAE of Peyer's patch and intestinal villi was immunohistochemically investigated in rat jejunoileum. As a result, cleaved caspase-3 was limited to several epithelial cells at the tip of FAE, whereas almost all of the epithelial cells were cleaved caspase-3 positive in intestinal villi. Cleaved caspase-9 was detected only in a few exfoliating or exfoliated epithelial cells of both FAE and intestinal villi. Nuclear DNA-fragmentation was detected only in several epithelial cells of the tip of FAE, while it was expressed from the middle regions in the intestinal villi. The DNase I expression of the epithelial cytoplasm was much weaker in FAE than in intestinal villi. Bcl-x expression was restricted in the apical cytoplasms of epithelial cells in the FAE, whereas it was restricted in whole cytoplasms in villous epithelial cells. These findings suggest that the progression of the apoptotic process in the epithelial cells of FAE is later than in the intestinal villi, so that the possibility of epithelial differentiation might be remained in the FAE, unlike in the intestinal villi.     

Comparison of the characters of the plaque-purified viruses from foot-and-mouth disease virus O/JPN/2000

At least two biotypes were observed at the 2nd passage stage after the isolation of Foot-and-mouth disease Virus (FMDV) O/JPN/2000 strain. These 2 types of viruses differed from their plaque phenotypes and were distinguishable by using a monoclonal antibody (MAb) 64G8 that was made for the FMDV O/JPN/2000 strain. One of these 2 biotypes formed small plaque (SP) and with immuno staining showed a positive reaction to MAb 64G8, while the other formed clear large plaque (LP) and did not react with MAb 64G8. The amino acid sequences of the capsid coding region (VP1-VP4) of the SP virus (SPV) and the LP virus (LPV) revealed two substitutions on the 133rd amino acid in VP2, and the 56th amino acid in VP3. These amino acid changes of SPV and LPV are Asn to Asp, Arg to His, respectively. The Arg of the 56th amino acid in VP3 that have been known as critical position of cell culture adapted virus. Only LPV showed high pathogenicity in suckling mice, and its LD(50) was calculated to be about 10(2) TCID(50)/0.1 ml. These results showed that the SPV that existed at the 2nd passage stage from isolation was a low virulence virus, which may suggest why the pathogenicity of O/JPN/2000 did not show clear symptoms in infected cattle.     

Quantitative structure-activity studies of benzoylphenylurea larvicides: I. Effect of substituents at aniline moiety against Chilo suppressalis walker

The larvicidal activity of a series of N-2,6-difluoro- and N-2,6-dichlorobenzoyl-N′-(4-substituted phenyl)ureas against nondiapause larvae of the rice stem borer, Chilo suppressalis Walker, was measured by topical application and oral administration methods under conditions with and without piperonyl butoxide as an inhibitor of oxidative metabolism. The effects of substituents at the anilide moiety on the larvicidal activity were analyzed quantitatively using physicochemical substituent parameters and regression analysis. The results indicate that the oxidative metabolism in the larval body which is favored by electron-donating substituents is significant in determining the activity. The activity, when the metabolic factor is eliminated, is enhanced by electron-with-drawing and hydrophobic substituents and lowered by bulky groups. The inhibitory activity against new cuticle formation of the same series of compounds was also measured using cultured integument of the rice stem borer diapause larva. The comparison of the quantitative analyses between larvicidal and integument-level activities shows that inhibition of cuticular development is the most important factor governing larvicidal activity.     

Effects of cyclodextrin-iodopropane complex on methane production, ruminal fermentation and microbes, digestibility and blood metabolites in steers

The effects when adding cyclodextrin‐iodopropane complex (CD‐IP) to a diet, on ruminal fermentation and microbes, digestibility, blood metabolites and methane production, were evaluated using four Holstein steers in a cross‐over design. The steers were fed Sudangrass hay plus concentrate mixture at a ratio 1.5:1, and CD‐IP (1% of dry matter) was given twice daily by mixing with concentrate mixture. Rumen and blood samples were collected at 0, 2, and 5 h after morning dosing. Ruminal pH and numbers of protozoa were unaffected by CD‐IP treatment. Ruminal molar proportion of acetate was decreased (P < 0.05), and propionate was increased (P < 0.01) at 2 h after CD‐IP dosing. Proportion of butyrate was increased (P < 0.05) and ammonia‐N was decreased (P < 0.05) at 2 and 5 h after CD‐IP dosing. Adding CD‐IP had no effect on the feed intake and digestion of nutrients. Plasma glucose was increased and urea‐N was decreased (P < 0.05) at 2 and 5 h after CD‐IP dosing. Methane production was decreased (P < 0.05) by approximately 18% in the treatment steers. Numbers of methanogenic bacteria were decreased (P < 0.05), while total viable counts, cellulolytic, sulfate reducing and acetogenic bacteria were unaffected. The present results are the first to show that CD‐IP can partially inhibit in vivo ruminal methanogenesis without adverse effects on digestion of nutrients.     

Ontogeny of testicular inhibin and activin in ducks: An immunocytochemical analysis

The ontogeny of testicular inhibin/activin in ducks was investigated. Testicular localization of three inhibin/activin subunits (α, β_A and β_B) was determined in embryonic and newly hatched ducks from 12 days of incubation to 1 day of age, in immature ducks and in adult ducks. In the duck embryonic testis, positive α‐subunit immunostaining was first detected in the Leydig cells and Sertoli cells on day 15 of incubation, whereas β_A‐subunit and β_B‐subunit immunostaining were found in Sertoli cells and primary germ cells on day 18 of incubation. In 1 month old ducks, intense staining of α‐subunit was present in the seminiferous epithelium consistent with localization in Sertoli cells and primary germ cells, and the immunostaining of the β_A‐ and β_B‐subunit was also present in Sertoli cells and primary germ cells. Specific immunostaining with inhibin/activin α‐, β_A‐ and β_B‐subunits antisera occurred in Sertoli cells in the adult duck testes. In conclusion, it was shown that, in the duck testis, the majority of α‐, β_A‐ and β_B‐subunits are colocalized in Sertoli cells with a certain degree of staining in germ cells and the α‐subunit is present in Leydig cells of embryonic testes before day 18 of incubation. These results indicate that Sertoli cells and possibly germ cells in the embryonic testes of late stage of incubation and newly hatched ducks, immature ducks and mature ducks may produce bioactive inhibin dimers, inhibin A and inhibin B, as a possible regulator of follicle‐stimulating hormone secretion. Free inhibin/activin subunits and their dimers may also play an autocrine/paracrine role in the development of the testis and spermatogenesis. Furthermore, early onset of the α‐subunit in duck testes indicates that it may have an autocrine/paracrine effect on steroid hormones, which is important for sex differentiation.     

Microbiota of 'airag', 'tarag' and other kinds of fermented dairy products from nomad in Mongolia

The traditional fermented dairy products were collected from three nomadic families in Donto‐Govi prefecture in Mongolia (central Mongolia), and those microbiota were analyzed. These samples consist three of ‘airag’, two of ‘tarag’, two of ‘isgelen tarag’ and ‘qoormog’, and some cheeses. In airag, Lactobacillus (L.) helveticus, L. kefiri, and Saccharomyces (S.) dairensis were common, and L. paracasei, L. plantarum, L. farciminis, S. cerevisiae, Issachenkia (I.) orientalis, Kluyveromyces (K.) wickerhamii were also found. In tarag, isgelen tarag and qoormog, L. helveticus, L. kefiri, L. fermentum, L. paracasei and L. acetotolerance were found. L. delbrueckii subsp. bulgaricus was also found in one tarag and one qoormog samples. Randomly amplified polymorphic DNA (RAPD) analysis showed that there were diversity in each L. helveticus family and products, and there were common strains found in airag and tarag in the same family.     

TGF-β and TNF-α stimulate MMP-9 production in murine epidermal keratinocytes

Matrix metalloproteinases 2 and 9 (MMP‐2 and ‐9) are zinc‐dependent metalloenzymes and have gelatin‐degrading activity. Both MMP are known to be secreted by many types of cells and play important roles in several biological changes including tissue remodeling and wound healing. In the present study, a primary culture of murine epidermal keratinocytes was prepared and effects of transforming growth factor‐β (TGF‐β), tumor necrosis factor‐α (TNF‐α) and interferon‐γ (IFN‐γ) on expression of MMP‐2 and MMP‐9 by the keratinocytes was examined. Gelatin zymography revealed that murine epidermal keratinocytes secreted proenzyme forms of MMP‐2 and MMP‐9, but the active forms of both MMP were hardly detectable, indicating that in vitro autoactivation of these proenzymes did not occur. Both TGF‐β and TNF‐α stimulated MMP‐9 production in a dose‐dependent manner, but the MMP‐2 level was not changed. Interferon‐γ hardly affected production of MMP‐2 or MMP‐9. Ribonuclease protection assay demonstrated that TNF‐α increased the level of MMP‐9 mRNA 6‐fold compared to the control, whereas TGF‐β slightly up‐regulated it. These results suggest that expression of MMP‐9 could be regulated by several cytokines in murine epidermal keratinocytes.     

International Harmonization of Nomenclature and Diagnostic Criteria (INHAND) progress to date and future plans

The INHAND Proposal (International Harmonization of Nomenclature and Diagnostic Criteria for Lesions in Rats and Mice) has been operational since 2005. A Global Editorial Steering Committee (GESC) manages the overall objectives of the project and the development of harmonized terminology for each organ system is the responsibility of the Organ Working Groups (OWG), drawing upon experts from North America, Europe and Japan.Great progress has been made with 9 systems published to date – Respiratory, Hepatobiliary, Urinary, Central/Peripheral Nervous Systems, Male Reproductive and Mammary, Zymbals, Clitoral and Preputial Glands in Toxicologic Pathology and the Integument and Soft Tissue and Female Reproductive System in the Journal of Toxicologic Pathology as supplements and on a web site – www.goreni.org. INHAND nomenclature guides offer diagnostic criteria and guidelines for recording lesions observed in rodent toxicity and carcinogenicity studies. The guides provide representative photo-micrographs of morphologic changes, information regarding pathogenesis, and key references. During 2012, INHAND GESC representatives attended meetings with representatives of the FDA Center for Drug Evaluation and Research (CDER), Clinical Data Interchange Standards Consortium (CDISC), and the National Cancer Institute (NCI) Enterprise Vocabulary Services (EVS) to begin incorporation of INHAND terminology as preferred terminology for SEND (Standard for Exchange of Nonclinical Data) submissions to the FDA. The interest in utilizing the INHAND nomenclature, based on input from industry and government toxicologists as well as information technology specialists, suggests that there will be wide acceptance of this nomenclature. The purpose of this publication is to provide an update on the progress of INHAND.