Transmission of porcine reproductive and respiratory syndrome virus from persistently infected sows to contact controls.

The objective of this study was to determine if porcine reproductive and respiratory syndrome virus (PRRSV) could persist in non-pregnant sows and if persistently infected sows could transmit virus to naive contact controls. Twelve PRRSV-naive, non-pregnant sows (index sows) were infected with a field isolate of PRRSV and housed in individual isolation rooms for 42 to 56 days postinfection. Following this period, 1 naive contact sow was placed in each room divided by a gate allowing nose-to-nose contact with a single index sow. Index sows were not viremic at the time of contact sow entry. Virus nucleic acid was detected by polymerase chain reaction, and infectious virus was detected by virus isolation in sera from 3 of the 12 contact sows at 49, 56, and 86 days postinfection. All 3 infected contacts developed PRRSV antibodies. Virus nucleic acid was detected in tissues of all of the 12 index sows at 72 or 86 days postinfection. Nucleic acid sequencing indicated that representative samples from index and infected contacts were homologous (> 99%) to the PRRSV used to infect index sows at the onset of the study. This study demonstrates that PRRSV can persist in sows and that persistently infected sows can transmit virus to naive contact animals.     

Differences in oxygen and carbon stable isotope ratios between hatchery and wild pink salmon fry

We examined differences in otolith oxygen (δ¹⁸O_otolith) and carbon (δ¹³C_otolith) stable isotope ratios between hatchery and wild pink salmon fry. The δ¹⁸O_otolith values of hatchery and wild fry were ?7.7 ± 0.2 ‰ and ?8.3 ± 0.3 ‰ (1σ), respectively. This difference reflected differences in temperature conditions experienced by each fry. The δ¹³C_otolith values of hatchery and wild fry were ?19.2 ± 0.3 ‰ and ?11.1 ± 1.8 ‰ (1σ), respectively. The lower δ¹³C_otolith values of hatchery fry were probably related to their intake of artificial diets. Discriminant analysis of δ¹⁸O_otolith and δ¹³C_otolith values demonstrated a highly significant difference between hatchery and wild fry with 95.8 % classification accuracy. Therefore, analysis of δ¹⁸O_otolith and δ¹³C_otolith precipitated in the fry stage may be useful for discriminating the origin of returning adult pink salmon. The discrimination method for returning adult fish would provide important information for evaluating the effect of hatchery release and their impact on the wild population.     

Migration history of Sakhalin taimen <Emphasis Type="Italic">Hucho perryi</Emphasis> captured in the Sea of Okhotsk,northern Japan,using otolith Sr:Ca ratios

Sakhalin taimen Hucho perryi populations have decreased drastically to near extinction. It is urgent to establish an effective conservation strategy based on an understanding of the characteristics of migration and habitat use of this species. We examined the migration history of anadromous Sakhalin taimen captured off the Sarufutsu coast, northern Hokkaido, Japan, using otolith Sr:Ca ratios and also examined the relationship between their otolith Sr:Ca ratios during freshwater and seawater residence in a rearing experiment. Otolith Sr:Ca ratios of some fish from the Sarufutsu coast showed freshwater levels (0.5–4.0 × 10⁻³) near the core, which thereafter increased to brackish water levels (4.0–6.0 × 10⁻³), and then to seawater levels (6.0–10.0 × 10⁻³) in the outermost regions. Those findings indicate that specimens from the Sarufutsu coast migrated to the brackish water region or the sea and spent most of their lives there. The anadromous migration pattern including the timing of downstream migration seems to be flexible among individuals in the species. They migrate between freshwater and seawater or brackish water several times during their lives, showing extensive habitat use. It is essential to secure the continuity among the freshwater, brackish water, and seawater areas for their effective conservation.     

Relative contributions from exposed inshore and estuarine nursery grounds to the recruitment of stone flounder, Platichthys bicoloratus, estimated using otolith Sr:Ca ratios

In Sendai Bay, stone flounder larvae settle and spend their juvenile period in either shallow exposed inshore nursery grounds or estuarine nursery grounds. The purpose of this study is to examine the relative contributions of these two kinds of nursery grounds to the flounder population using otolith strontium:calcium ratios. Stone flounder juveniles were collected from both nursery grounds, and one- and two-year-old flounder were caught deeper in Sendai Bay. Sr and Ca content in the otoliths were measured by electron probe micro analysis. The Sr:Ca ratios in the otolith section corresponding to the early postsettlement period ranged from 3.06 to 3.85 for the exposed inshore areas with stable low temperature and high salinity conditions, and from 3.81 to 5.32 in brackish estuaries with high temperature and low salinity conditions but with large diel and tidal cyclical fluctuations. Values from an estuarine site with stable salinity ranged from 3.58 to 4.15 overlapping with both the above ranges. Rearing experiments supported our inference that the high otolith Sr:Ca ratios of juveniles inhabiting estuarine nursery grounds are attributable to higher temperature and physiological stress caused by the large diel temperature and salinity fluctuations within the estuaries. Estimation of the Sr:Ca ratio of recruited fish using the otolith section formed while in the nursery area showed that at least 20 out of 42 individuals examined originated from estuarine nursery grounds. The present study indicates that estuaries play an important role as nursery grounds for stone flounder, producing about half of the stock in spite of the small and restricted area compared with the wide expanse of the exposed inshore area.     

Effect of calcium concentration in nutrient solution before and after inoculation with Ralstonia solanacearum on resistance of tomato seedlings to bacterial wilt

We previously reported that calcium (Ca) nutrition in tomato (Lycopersicon esculentum Mill.) significantly affected the resistance to bacterial wilt caused by Ralstonia solanacearum Smith. To elucidate the mechanisms underlying the Ca-dependent resistance, the effect of the Ca concentration in the nutrient solution applied before and after inoculation with the pathogen on the resistance of tomato seedlings to bacterial wilt was studied. One week before inoculation, seedlings were transferred to nutrient solutions containing Ca at concentrations of 0.4, 4.4, or 20.4 mM. Soon after inoculation, the seedlings that were treated with each concentration of Ca before inoculation were transferred to solutions containing the same three concentrations of Ca. Although the disease development was not affected by the concentration of Ca in the solution before inoculation, a higher concentration of Ca after inoculation reduced the disease severity. This result suggests that the concentration of Ca in the host, especially in the cell walls, before infection may not be directly involved in the Ca-dependent resistance of tomato seedlings to bacterial wilt.     

Development of a high-throughput screening method using a cell-based, lawn format assay for the identification of novel plant defense activators from combinatorial peptide libraries

Plants respond to attack by pathogens through various defense mechanisms. These defense responses are triggered by a variety of molecules derived from pathogenic microorganisms as well as host plants. In this study, we developed a high-throughput screening method using a cell-based lawn format assay for the identification of novel peptides that can induce plant defense responses from combinatorial peptide libraries. Solid-phase peptide libraries were synthesized using a photocleavable linker and immobilized using agarose gel. The peptides were partially cleaved from beads, and the agarose gel was layered on the tobacco cells. The defense response was then observed by detecting the generated H2O2 using a sensitive H2O2 indicator dye, N-(carboxymethylaminocarbonyl)-4,4'-bis(dimethylamino)diphenylamine sodium salt (DA-64). Using this assay format, a 6859-member peptide library based on the sequence of flagellin-derived peptides was screened, and several structural features important for the activity were obtained.     

Potential use of otolith microchemistry for field studies of temperature-dependent sex determination and gonadal degeneration in fish

This paper describes a novel approach to study the expression of temperature-related reproductive phenomena such as thermolabile sex determination (TSD) and heat-induced germ cell loss in wild fish populations. The proposed approach is based on the reconstruction of the past thermal history of individual fish through the microchemical analysis of the otoliths (mineralized structures responsible for the sense of balance). As an example, we outline the preliminary results of an investigation on the natural occurrence of TSD in pejerrey Odontesthes bonariensis, an atherinid fish in which all-female to all-male populations can be produced by experimental manipulation of the rearing temperature during the critical time of sex differentiation, from Lake Kasumigaura, Japan.     

Mechanical transmission of porcine reproductive and respiratory syndrome virus throughout a coordinated sequence of events during cold weather

Using a field-based model, mechanical transmission of porcine reproductive and respiratory syndrome virus (PRRSV) was assessed throughout a coordinated sequence of events that replicated common farm worker behavior during cold weather (< 0°C). The model involved fomites (boots and containers), vehicle sanitation, transport, and the movement of personnel. A field strain of PRRSV was inoculated into carriers consisting of snow and water, and carriers were adhered to the undercarriage of a vehicle. The vehicle was driven approximately 50 km to a commercial truck washing facility where the driver's boots contacted the carriers during washing, introducing the virus to the vehicle interior. The vehicle was then driven 50 km to a simulated farm site, and the driver's boots mechanically spread virus into the farm anteroom. Types of containers frequently employed in swine farms (styrofoam semen cooler, metal toolbox, plastic lunch pail, and cardboard animal health product shipping parcel) contacted drippings from footwear on the anteroom floor. The truck wash floor, vehicle cab floor mats, boot soles, anteroom floor, and the ventral surface of containers were sampled to track the virus throughout the model. Ten replicates were conducted, along with sham-inoculated controls. At multiple sampling points PRRSV nucleic acid was detected in 8 of 10 replicates. In each of the 8 PCR-positive replicates, infectious PRRSV was detected on the surfaces of containers by virus isolation or swine bioassay. All sham-inoculated controls were negative. These results indicate that mechanical transmission of PRRSV can occur during coordinated sequence of events in cold weather.