Services

Diagnostics, experimental trials, training and field advisory services

Nematode diagnostics

Extractions are carried out according to recognised official methods, described in EPPO Bulletin 43.

Light micrograph of Xiphinema index
Xiphinema index, adult female. Vineyard soil, Puglia, Italy.

Nematofauna analysis

The report is not limited to counting the parasites. It sets out the composition of the entire soil nematode community, broken down by trophic role — parasites, predators, omnivores and decomposers — together with the calculation of the parasite/predator ratio.

Chart showing the trophic composition of the soil nematode community
Left: trophic composition of the soil nematode community — parasites, predators, omnivores and decomposers — expressed as a percentage of the total individuals extracted.
Chart showing the distribution of plant-parasitic nematodes by genus
Right: distribution of plant-parasitic nematodes by genus, expressed as a percentage of their total.

This picture provides an indicator of the biological status of the soil that counting plant parasites alone cannot give. It is useful in planning replanting, in assessing the effect of treatments on the beneficial soil community, and in monitoring integrated or organic management systems.

Nematicide efficacy trials

In vitro experimental work is carried out at the Horto Service laboratory. Trials in growth chambers and under glasshouse or field conditions are carried out in close cooperation with the AgroService Test Centre in Andria (Apulia, Italy).

How a trial is designed

The design of the study is determined by the expected efficacy of the formulation under test. It governs the choice of reference product, the experimental design, the crop, the duration of the trial expressed as the number of generations of the parasite, and the timing of the intermediate nematological assessments.

Experimental plots for nematicide efficacy trials
Left: experimental plots for the efficacy assessment of nematicides against Meloidogyne spp. on tobacco (Nicotiana tabacum) in open field. Right: bench trial with artificial inoculum at a known density of Meloidogyne spp. on tomato (Solanum lycopersicum). The use of sterilised substrate and calibrated inoculum allows a large number of treatments to be compared under controlled conditions.

A highly effective formulation requires stressful conditions, high parasite pressure, a fumigant reference product and assessment over two to four generations, on long-cycle crops. A formulation of modest efficacy is assessed over one to two generations, with a liquid contact reference product and limited pressure, on short-cycle crops.

The importance of an intermediate assessment

The high biotic potential of root-knot nematodes leads to rapid population build-up. Within two to three generations the waves of infection saturate the feeding sites and the infestation tends to level off at the highest values observable in the field, irrespective of the initial differences between treatments.

As a result, the initial efficacy of control is progressively masked. An assessment carried out too long after transplanting therefore risks failing to discriminate between genuinely different treatments: for this reason an intermediate assessment, particularly on roots, may prove more informative than the final one, and makes it possible to establish for how long the crop actually remained protected.

Selection and management of trial sites

Application of the formulations

For chemigation trials a dedicated plot-level irrigation system is used, with the same layout and spacing as the farm system, laid out on prepared and levelled ground. Each plot has its own manifold, left in place for subsequent applications.

Pressure-compensating low-flow emitters are used, to promote horizontal wetting of the soil. The volumes of water allocated to each plot are pre-measured, and delivery is fractionated and slow, so as to avoid surface run-off and deep drainage. The water status of the soil is checked before application, with pre-wetting on the previous day where necessary.

In vitro and controlled-environment testing

The activity of a substance can be verified at scales progressively closer to real conditions, before committing resources to a field trial. Horto Service carries out the entire sequence.

Preliminary in vitro assays

Determination of the lethal doses LD50 and LD90 on infective juvenile stages. Egg masses are collected from infected roots and incubated in distilled water in a growth chamber at 25 °C. The emerged J2 juveniles are collected and divided into calibrated aliquots, to which the treatments are applied at the concentrations under test. Studies of this kind serve to determine the nematicidal and nematostatic potential of a substance, as well as the persistence of its efficacy after exposure has ended. Hatching tests are also carried out directly on the egg masses.

Preliminary pot assays with artificial inoculum

Trials on inert sand or steam-sterilised sandy soil, which place no limit on the number of treatments that can be compared. The inoculum is obtained by mass extraction of eggs from infected roots, divided into calibrated aliquots and applied in suspension at a known density.

Treatments are applied as a drench at limited flow rate, with pre-wetting and reproduction of the chemigation stages. Assessments cover biometric parameters, gall index and reproduction rate of the parasite at the end of the first and second generation.

Inoculum production

The laboratory produces inoculum of Meloidogyne spp. by mass extraction of eggs from infected roots. The availability of calibrated inoculum is the precondition for reproducible assays, and also makes it possible to even out the infestation at field trial sites where the natural distribution is uneven.

Production of Meloidogyne spp. inoculum in the laboratory
Left: mass extraction of adult females of Meloidogyne spp. from infected root systems. Centre: suspension of eggs, second-stage juveniles (J2) and adult males of Meloidogyne spp. obtained from the egg masses. Right: tube with a sediment of eggs intended for the inoculation of experimental units.

Training

Horto Service designs and delivers technical training courses in agricultural nematology for trial technicians.

The content covers the biology of plant-parasitic nematodes, population dynamics and interaction with the environment, sampling criteria for soil and plant matrices, the principles of integrated control, and the design of experimental trials in accordance with Good Experimental Practice.

Technical advisory services

Horto Service also operates internationally, providing technical assistance, training of local technicians and field diagnostics.

For non-nematological analyses — microbiological, chemical or physical — the laboratory relies on external facilities.

Composition of micrographs of plant-parasitic nematodes