Transmission of extended-spectrum cephalosporin-resistant Salmonella harboring a blaCMY-2-carrying IncA/C2 plasmid chromosomally integrated by ISEcp1 or IS26 in layer breeding chains in Japan

Dissemination of extended-spectrum cephalosporin (ESC)-resistant Salmonella is a public health concern in the egg production industry. ESC-resistant Salmonella often acquires the bla gene via insertion sequences (ISs). Therefore, this study aimed to assess antimicrobial resistance in Salmonella from Japanese layer breeding chains and egg processing chains, and determine the genetic profiles of IS-like elements in ESC-resistant Salmonella. Antimicrobial susceptibility testing was performed on 224 isolates from 49 facilities involving layer breeder farms, hatcheries, pullet-rearing farms, and layer farms in breeding chains along with egg processing chains. ESC-resistant Salmonella strains were whole-genome sequenced. Among them, 40 (17.9%) were resistant to at least streptomycin, tetracycline, ampicillin, chloramphenicol, cefpodoxime, nalidixic acid, ciprofloxacin, and/or kanamycin despite lacking resistance to azithromycin and meropenem. Moreover, 15 were ESC-resistant Salmonella harboring bla_CMY-2 (Salmonella enterica serovar Ohio, n=12; S. Braenderup, n=1; untypeable with O7:b:-, n=1) and bla_CTX-M-14 (S. Cerro, n=1). IncA/C₂ plasmids containing ISEcp1, IS26, and multiple antimicrobial resistance genes (including bla_CMY-2) were identified in S. Ohio isolates from pullet-rearing and layer farms belonging to the same company. Chromosomal integration of partial or whole IncA/C₂ plasmids was seen with two S. Ohio isolates via ISEcp1 or IS26, respectively. Antimicrobial resistance genes such as bla_CMY-2 might be transmitted among the upper and the lower levels of layer breeding chains via the replicon type IncA/C₂ plasmids containing ISEcp1 and IS26.     

Nitrate Decrease with Isotopic Fractionation In Riverside Sediment Column During Infiltration Experiment

This study determined isotopic fractionation of nitrate-nitrogen during denitrification in riparian aquifer sediments by sequential-column experiments at two different water flow rates. The purpose was to discuss the relationships of nitrogen isotopic enrichment factor of denitrification, denitrification rate, and infiltrating condition in sediment. Sandy sediment and groundwater were collected from floodplain in the middle reach of the Tama River where nitrogen isotopic enrichment factor for denitrification had been measured in a result of previous field observation. The denitrification rates and the isotopic enrichment factors of nitrate-nitrogen were estimated at 2.1 mgN dry-kg^?1 day^?1 and ?32.9‰ during the low water flow condition, and at 3.5 mgN dry-kg^?1 day^?1 and ?34.1‰ during the high water flow condition. The calculated water flow rates of the present column experiments were 300–400 times higher than that of the field observation. Because of the fast flow rate, nitrate was expected to continuously pass though active denitrifying sites in the small pore spaces of sediment, and nitrate supply to denitrifers by infiltration flow transport greatly exceeded the supply by diffusion. The observed denitrification rates was proportional to Peclet number as the ratio of solute nitrate transport to the diffusion, and apparent nitrogen isotopic enrichment factors during denitrification of our column experiments were lower than those obtained from the field observation. This study showed that denitrification rate and apparent nitrogen isotopic enrichment factor of denitrification in sediment depended on Peclet number.     

Effect of Biodegradable Gelatin β-Tri Calcium Phosphate Sponges Containing Mesenchymal Stem Cells and Bone Morphogenetic Protein-2 on Equine Bone Defect

This study evaluated the osteoanagenetic effects of administering biodegradable gelatin β-tri calcium phosphate sponges containing mesenchymal stem cells (MSCs) and bone morphogenetic protein-2 (BMP-2) on equine bone defect. Six healthy Thoroughbred horses were used in this study. Horses were anesthetized, and skin incisions were made on all the limbs. Splint bones were exposed and a 1-cm bone defect was created in each exposed bone. Gelatin β-tri calcium phosphate sponges containing MSC and BMP-2 (MSC + BMP-2 sponge), MSC only (MSC sponge), BMP-2 only (BMP-2 sponge), or saline (saline sponge) were implanted into each bone defect at random. Defects were monitored for 16 weeks by radiography followed by computed tomography (CT) and histologic analyses. At 16 weeks, radiographic scores of MSC + BMP-2 sponge-treated defects were significantly higher than those of saline-treated defects (P = .027). Moreover, the CT value of the MSC + BMP-2 sponge group was significantly higher than that of the other groups (P = .027; P = .046; and P = .027, respectively), and the histologic score of the MSC + BMP-2 sponge group was significantly greater than that of the saline sponge group (P = .041). We conclude that MSC + BMP-2 sponge administration to bone defects accelerates bone regeneration in equines.     

Effects of Full-Length Kisspeptin Administration on Follicular Development in Japanese Black Beef Cows

Kisspeptin is a key molecule that stimulates gonadotropin secretion via release of gonadotropin-releasing hormone (GnRH). In the present study, our aim was to investigate whether kisspeptin has stimulatory effects on follicular development via GnRH/gonadotropin secretion in cows. Japanese Black beef cows were intravenously injected with full-length bovine kisspeptin [Kp-53 (0.2 or 2 nmol/kg)] or vehicle 5 days after they exhibited standing estrus (Day 0). In cows injected with Kp-53 at 2 nmol/kg, the follicular sizes of the first dominant follicles increased on Day 6 and thereafter. Ovulation of the first dominant follicle occurred in 1 out of 4 cows treated with Kp-53 at 2 nmol/kg. Injection of Kp-53 at 2 nmol/kg increased the concentration of plasma luteinizing hormone (LH) but not follicle-stimulating hormone, over a 4-h period following injection in all cows. The present study suggests that administration of full-length kisspeptin causes LH secretion, which is sustained for a few hours, and it is capable of stimulating follicular development and/or ovulation.     

Effect of the number of tillages in fallow season and fertilizer type on greenhouse gas emission from a rice (Oryza sativa L.) paddy field in Ehime,southwestern Japan

Agricultural fields, including rice (Oryza sativa L.) paddy fields, constitute one of the major sources of atmospheric methane (CH₄) and nitrous oxide (N₂O). Organic matter application, such as straw and organic fertilizer, enhances CH₄ emission from paddy fields. In addition, rice straw management after harvest regulates CH₄ emissions in the growing season. The interaction of tillage times and organic fertilizer application on CH₄ and N₂O emissions is largely unknown. Therefore, we studied the effects of fallow-season tillage times and fertilizer types on CH₄ and N₂O emissions in paddy fields in Ehime, southwestern Japan. From November 2011 to October 2013, four treatments, two (autumn and spring) or one (spring) in the first year, or two (autumn and spring) or three (autumn, winter, and spring) in the second year times of tillage with chemical or organic fertilizer application, were established. Gas fluxes were measured by the closed-chamber method. Increasing the number of tillage times from one to two decreased succeeding CH₄ emission and the emission factor for CH₄ (EF_CH4) in the rice-growing season, suggesting that the substrate for CH₄ production was reduced by autumn and spring tillage in the fallow season. Higher EF_CH4 [1.8–2.0 kg carbon (C) ha^?1 d^?1] was observed when more straw was applied (6.9–7.2 Mg ha^?1) in the second year. Organic fertilizer application induced higher CH₄ emission just after the application as basal and supplemental fertilizers, especially at a lower straw application rate. This indicated that EF_CH4 in the organically managed fields should be determined individually. Organic fertilizer application with two tillage times induced N₂O efflux during the rice-growing season in the second year, but N₂O emissions were not affected by winter tillage. Although paddy fields can act as an N₂O sink because of reduced soil conditions when straw application was high, application of organic C and nitrogen as fertilizer can enhance N₂O production by the denitrification process during the growing season, especially in the ripening stage when soil anaerobic conditions became moderate. These results suggest that negative emission factors for N₂O (EF_N2O) can be applied, and EF_N2O of organic fertilizer should be considered during the estimation of N₂O emission in the paddy field.     

Nutritional Study of Silicon in Graminaceous Crops (Part 2)

It was reported in the previous paper₁₎ that rice plants showed retardation of their vegetative growth and decrease of degree of seed setting when their silicon content was extremely low. It was concluded from these facts that silicon might most probably be essential for rice. But physiological functions or behaviours of silicon in plants should be clarified in order to solve this problem completely. Uptake and disttibution of silicon in rice plants were first investigated here, especially compared with phosphorus. A possibility of using silicon radioactive isotope (₃₁Si) for this kind of experiment was also investigated. Although tracer techniques have been greatly developed in elucidating behaviours or physiological functions of various elements in plants, radioactive silicon isotopes have been used very little in this field of science because of this very short half lives. Recently Rothbuhr and Scott ²⁾ reported having used radioactive silicon isotope for uptake experiments by plants in Harwell, England. Since a nuclear reactor (JRR-I) in the Japanese Atomic Energy Research Institute started to work in 1959, radioactive isotopes which have rather short half lives have become available for plant experiments in Japan. Methods of preparation of ³¹Si for plant experiments were examined and by using ³¹Si it was studied how silicon was absorbed and translocated by rice and wheat under some particular environmental conditions, and also how the uptake and distribution of it in rice plants were affected by various kinds of metabolic inhibitors.     

Accumulation of Zinc and Copper in an Arable Field after Animal Manure Application

An experiment was conducted to examine the accumulation and mobility of heavy metals (Zn and Cu) at different depths in three types of arable soils (Brown Lowland soil, Andosol, and Brown Forest soil) amended with cattle and pig farmyard manures for 5 years. Nitric-perchloric acid digestion was performed for the determination of the total amounts of heavy metals, and 0.1 M hydrochloric acid extraction was performed for the determination of the amounts of soluble heavy metals. Results of the soil analysis indicated that pig farmyard manure application resulted in serious contamination of arable soils with Zn and potentially Cu. Especially, the Brown Forest soil displayed a high ability to accumulate heavy metals on the soil surface. Total-Zn concentration in surface soils was considerably affected by the holding capacity of soluble-Zn traction. Although the Andosol amended with pig farmyard manure showed higher concentrations of heavy metals related to the higher ability of retention on a weight basis, the soil did not contribute to high heavy metal accumulation because of its low bulk density. Heavy metals were easily leached in sandy soils such as Brown Lowland soil, and Cu was potentially stable compared with Zn. We suggest that long-term pig farmyard manure application to the Brown Lowland soil and Andosol with a light soil texture is associated with a higher risk of groundwater pollution than the application to the Brown Forest soil.     

Potato extract (Potein) suppresses food intake in rats through inhibition of luminal trypsin activity and direct stimulation of cholecystokinin secretion from enteroendocrine cells

Dietary proteins and trypsin inhibitors are known to stimulate the secretion of the satiety hormone cholecystokinin (CCK). A potato extract (Potein) contains 60% carbohydrate and 20% protein including trypsin inhibitory proteins. In this study, we examined whether Potein suppresses food intake in rats and whether it directly stimulates CCK secretion in enteroendocrine cells. In fasted rats, food consumption was measured up to 6 h after the oral administration of Potein or soybean trypsin inhibitor (SBTI). CCK-releasing activities of Potein and SBTI were examined in the murine CCK-producing cell line STC-1. Potein inhibited the trypsin activity in vitro with a potency 20-fold lower than that of SBTI. Oral administration of Potein dose-dependently suppressed food intake for 1-6 h. Potein, but not the SBTI, dose-dependently induced CCK secretion in STC-1 cells. These results suggest that Potein suppresses food intake through the CCK secretion induced by direct stimulation on enteroendocrine cells and through inhibition of luminal trypsin.