Multiplex real-time PCR for the detection and differentiation of equid herpesvirus 1 (EHV-1) and equid herpesvirus 4 (EHV-4).

A multiplex real-time PCR was designed to detect and differentiate equid herpesvirus 1 (EHV-1) and equid herpesvirus 4 (EHV-4). The PCR targets the glycoprotein B gene of EHV-1 and EHV-4. Primers and probes were specific to each equine herpesvirus type and can be used in monoplex or multiplex PCRs, allowing the differentiation of these two closely related members of the Alphaherpesvirinae. The two probes were minor-groove binding probes (MGB?) labelled with 6-carboxy-fluorescein (FAM?) and VIC® for detection of EHV-1 and EHV-4, respectively. Ten EHV-1 isolates, six EHV-1 positive clinical samples, one EHV-1 reference strain (EHV-1.438/77), three EHV-4 positive clinical samples, two EHV-4 isolates and one EHV-4 reference strain (EHV-4 405/76) were included in this study. EHV-1 isolates, clinical samples and the reference strain reacted in the EHV-1 real-time PCR but not in the EHV-4 real-time PCR and similarly EHV-4 clinical samples, isolates and the reference strain were positive in the EHV-4 real-time PCR but not in the EHV-1 real-time PCR. Other herpesviruses, such as EHV-2, EHV-3 and EHV-5 were all negative when tested using the multiplex real-time PCR. When bacterial pathogens and opportunistic pathogens were tested in the multiplex real-time PCR they did not react with either system. The multiplex PCR was shown to be sensitive and specific and is a useful tool for detection and differentiation of EHV-1 and EHV-4 in a single reaction. A comprehensive equine herpesvirus disease investigation procedure used in our laboratory is also outlined. This procedure describes the combination of alphaherpesvirus multiplex real-time PCR along with existing gel-based PCRs described by other authors.

Organic inputs, tillage and rotation practices influence soil health and suppressiveness to soilborne pests and pathogens of ginger.

A field experiment was established in which an amendment of poultry manure and sawdust (200 t/ha) was incorporated into some plots but not others and then a permanent pasture or a sequence of biomass-producing crops was grown with and without tillage, with all biomass being returned to the soil. After 4 years, soil C levels were highest in amended plots, particularly those that had been cropped using minimum tillage, and lowest in non-amended and fallowed plots, regardless of how they had been tilled. When ginger was planted, symphylans caused severe damage to all treatments, indicating that cropping, tillage and organic matter management practices commonly used to improve soil health are not necessarily effective for all crops or soils. During the rotational phase of the experiment, the development of suppressiveness to three key pathogens of ginger was monitored using bioassays. Results for root-knot nematode (Meloidogyne javanica) indicated that for the first 2 years, amended soil was more suppressive than non-amended soil from the same cropping and tillage treatment, whereas under pasture, the amendment only enhanced suppressiveness in the first year. Suppressiveness was generally associated with higher C levels and enhanced biological activity (as measured by the rate of fluorescein diacetate (FDA) hydrolysis and numbers of free-living nematodes). Reduced tillage also enhanced suppressiveness, as gall ratings and egg counts in the second and third years were usually significantly lower in cropped soils under minimum rather than conventional tillage. Additionally, soil that was not disturbed during the process of setting up bioassays was more suppressive than soil which had been gently mixed by hand. Results of bioassays with Fusarium oxysporum f. sp. zingiberi were too inconsistent to draw firm conclusions, but the severity of fusarium yellows was generally higher in fumigated fallow soil than in other treatments, with soil management practices having little impact on disease severity. With regard to Pythium myriotylum, biological factors capable of reducing rhizome rot were present, but were not effective enough to suppress the disease under environmental conditions that were ideal for disease development.

Integrating organic matter into banana plantation in North Queensland: the effects on soil properties

The major banana production areas in Australia are particularly sensitive to environments due to their close proximity to areas of World Heritage rainforest and the Great Barrier Reef catchment. Management of soil quality, nutrients and pesticides are vital to maintaining the integrity of these sensitive areas. Studies on cropping systems have suggested that integrating organic matter into ground cover management would improve the quality of soil under banana cultivation. In this study, an alternative management practice for bananas, which addresses the management of organic matter and fertiliser application, was assessed and compared to the conventional practice currently employed in the banana industry. Several chemical, physical and biological soil parameters were measured including: pH, electrical conductivity, water stable aggregates, bulk density, water filled pore space, porosity, water content, fluorescein diacetate hydrolyis (FDA) and beta-glucosidase activity. The alternative management practice did not have a significant impact of the production and growth of bananas but overall improved the quality of the soil. Although some differences were observed, the chemical and physical soil characteristics did not differ dramatically between the two management systems. The addition of organic matter resulted in the soil under alternative practice having higher FDA and beta-glucosidase levels, indicating higher microbial activity. The integration of organic matter into the management of bananas resulted in positive benefits on soil properties under bananas, however, methods of maintaining organic matter in the soil need to be further researched.

The Effect of Silica and Manure Addition into Suppressive and Conducive Soil on the Incidence of Fusarium Wilt Disease of Banana

Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense (Foc), is one of the most destructive diseases of banana. One potential method to manage fusarium wilt of banana is by manipulating the nutrient status in the soil. This study was conducted to determine the quality of Foc suppressive and conducive soil, the influence of soil application of silica and manure on the incidence of fusarium wilt of banana. Surveys were conducted in five banana plantations in three provinces in Indonesia: Lampung-Sumatra, West Java and Central Java. From the five locations, one location (Sala-man-Central Java) was heavily infected by Foc, another location (NTF Lampung-Sumatera) was slightly infected by Foc, while the rest (Sarampad-West Java, Talaga-West Java and GGP Lampung-Sumatra) were healthy banana plantations without Foc infection. Labile carbon analysis showed that the Foc suppressive soil had greater labile carbon content than conducive soil. Also, the analysis of fluorescein diacetate hydrolysis (FDA) and ?-glucosidase showed greater microbial activity in suppressive soil than the conducive soil. Observations of the incidence of necrotic rhizome of Foc susceptible 'Ambon Kuning' (AAA) banana cultivar showed that in the suppressive soil taken from Sarampad West Java, the application of silica and manure helped suppress fusarium wilt disease development. In the conducive soil taken from Salaman-Central Java, silica and manure applications were not able to suppress disease incidence. The result of this study indicated that in suppressive soil, the application of silica can increase plant resistance to Foc infection, while manure application can increase soil microbial activity, and suppress Foc development.