A review of pest surveillance techniques for detecting quarantine pests in Europe

This paper provides reviews of the most commonly used methods to detect plant pests belonging to groups of invasive organisms with high economic relevance, including Coleoptera (bark beetles, flathead borers, leaf beetles, longhorn beetles, weevils), Diptera (cone and seed flies, fruit flies), Homoptera (aphids, leafhoppers and psyllids, whiteflies), Lepidoptera (moths and butterflies), Thysanoptera (thrips), bacteria (potato brown rot Ralstonia solanacearum) and fungi (pitch canker disease Gibberella circinata, brown rot disease Monilinia fructicola). Future perspectives in detection methods are discussed, with particular reference to the considerable increase in the volume, commodity type and origins of trade in plant material from third countries, the introduction of new crops, the continuous expansion of the EU with new border countries being added, and the impact of climate change affecting the geographical boundaries of pests and their vectors.     

Optimization of in-house ELISA based on recombinant major sperm protein (rMSP) of Dictyocaulus viviparus for the detection of lungworm infection in cattle

The aim of this study was to optimize an in-house ELISA based on a recombinant version of the major sperm protein (MSP) of Dictyocaulus viviparus for routine diagnosis of lungworm infection in cattle. A recombinant MSP (rMSP) was cloned into pGEX-6P-1 vector and expressed as a glutathione-S-transferase (GST) fusion protein in Escherichia coli BL21 (DE3) chemically competent cells. The product was then employed as capture antigen in an ELISA, and validated against 304 samples of known status (216 negative and 88 positive) in which the antibody levels in sera had also been measured earlier with a commercial ELISA kit (Ceditest® lungworm ELISA). The receiver operating characteristic (ROC) curve analysis of the ELISA results estimated the optimized diagnostic sensitivity and specificity as 97.7% (95% confidence interval [CI]: 91.9–99.7%) and 98.1% (CI: 95.3–99.5%), respectively. The results from the in-house rMSP-based ELISA were compared with results obtained on both fecal examination and the Ceditest® lungworm ELISA. Rising antibody levels in sera of experimentally infected calves were observed between 21 and 28 days post infection, when patency was also confirmed by the presence of larvae in feces. Notably, using the in-house rMSP-based ELISA infection was confirmed in calves shedding larvae approximately 3–4 weeks post inoculation, while using the Ceditest® lungworm ELISA those animals remained negative. Additionally, 251 sera samples from calves naturally exposed to the parasites on pasture were used to evaluate the test. In in-house rMSP-based ELISA no cross-reactions were observed with sera from calves infected with the gastrointestinal nematodes (Ostertagia ostertagi and Cooperia oncophora), even though the presence of eggs in the feces was confirmed. Overall, the in-house rMSP-based ELISA optimized in this study showed excellent diagnostic performance for detection of lungworm infection in cattle.     

Stimulation of tree defenses by Ophiostomatoid fungi can explain attack success of bark beetles on conifers

? Our aim is to present why the hypothesis, that Ophiostomatoid fungi play an important role in the establishment of most bark beetle species on living conifers, is valuable.

? After summarizing knowledge about the relationships of bark beetles with conifers and fungi, we conclude that controversy results from misinterpretations when using fungal pathogenicity to demonstrate the role of Ophiostomatoid fungi in beetle establishment on host trees.

? We demonstrate that fungal pathogenicity is not the right parameter to appreciate the role of fungus in beetle establishment on host trees. We argue that artificial low density inoculations that allow the appreciation of fungus ability to stimulate tree defenses and thus to help beetles in overcoming tree resistance must be used in complement to mass inoculations. In both cases, results must be expressed in terms of tree defense stimulation rather than in terms of tree killing.

1. Fungal species stimulating tree defenses are generally not those that grow the best in the sapwood.
2. We argue that beetle development in the phloem, fungal invasion of the sapwood and phloem, and tree death, occur after tree defenses are exhausted, and that any fungus present in the beetle gallery could thus potentially invade the sapwood after defense exhaustion.

? We conclude that stimulation of the tree defense reactions in both the phloem and the superficial sapwood is a real benefit brought by fungi to the beetles during the first phase of establishment (overcoming tree resistance).

? Considering the origin of the bark beetle fungus associations attacking living trees and their general functioning based on stimulation of tree defenses, we develop three hypotheses:

1. any beetle species would be helped in its establishment in a given tree species by developing an association, even loosely, with a fungus species belonging to the Ophiostomatoid flora of that tree species;
2. the necessity of a considerably low level of tree resistance for fungus extension into the tree is the selection pressure that has led fungi to develop their intrinsic ability to stimulate tree defenses, through their ability to grow into the phloem. This association can be completed by antagonistic fungal species controlling extension of the previous fungal species in the tree tissues;
3. Beetle species using the strategy of overcoming tree resistance are associated with a fungal complex, of which species could assume three roles regarding relationships between beetles and trees: 1- to stimulate tree defenses in the phloem and superficial sapwood, 2- to grow into the sapwood after tree resistance is overcome, and 3- to control phloem extension of the first other two categories. Bringing nutrients to the beetle progeny can be a fourth role.

? We propose that bark beetle — Ophiostomatoid associations can be categorized, based on associations’ frequency and complexity while taking into account beetle aggressiveness. We show that a close correspondence exists between beetles’ aggressiveness and the ability of their main associated fungal species to stimulate the defenses of their host tree.

? We conclude with suggesting that most sapwood invading fungi might be “cheaters” which have taken advantage of the efficiency of the relationship between beetles and fungi that stimulate tree defenses.

     

Polybrominated Diphenyl Ether Dynamics in Ambient Air and Atmospheric Bulk/Wet Deposition in Downtown Paris (France)

This study reports on the polybrominated diphenyl ether (PBDE) atmospheric dynamics in the dense urban environment of downtown Paris (France). Eight PBDE congeners (BDE-28, BDE-47, BDE-100, BDE-99, BDE-154, BDE-153, BDE-183 and BDE-209) were simultaneously analysed in bulk and wet atmospheric deposition, as well as in ambient air (gaseous/particulate phases), twice a month over a 12-month period (May 2008–May 2009). The total air concentration of Σ 8PBDEs ranged between 12 and 185 pg m⁻³, and the seasonal variations of PBDE levels were controlled by air temperature (except for BDE-209). Regarding bulk deposition, Σ 8PBDE flux was in the range 5–94 ng m⁻² day⁻¹, and it was positively correlated with atmospheric particulate phase PBDE concentrations. Meanwhile, the measured wet deposition flux of Σ 8PBDE ranged between 2 and 24 ng m⁻² day⁻¹, and its median contribution to bulk deposition was 39% only, which highlights the importance of dry deposition.     

Antigenic and genetic variations in European and North American equine influenza virus strains (H3N8) isolated from 2006 to 2007

Equine influenza virus (EIV) surveillance is important in the management of equine influenza. It provides data on circulating and newly emerging strains for vaccine strain selection. To this end, antigenic characterisation by haemaggluttination inhibition (HI) assay and phylogenetic analysis was carried out on 28 EIV strains isolated in North America and Europe during 2006 and 2007. In the UK, 20 viruses were isolated from 28 nasopharyngeal swabs that tested positive by enzyme-linked immunosorbent assay. All except two of the UK viruses were characterised as members of the Florida sublineage with similarity to A/eq/Newmarket/5/03 (clade 2). One isolate, A/eq/Cheshire/1/06, was characterised as an American lineage strain similar to viruses isolated up to 10 years earlier. A second isolate, A/eq/Lincolnshire/1/07 was characterised as a member of the Florida sublineage (clade 1) with similarity to A/eq/Wisconsin/03. Furthermore, A/eq/Lincolnshire/1/06 was a member of the Florida sublineage (clade 2) by haemagglutinin (HA) gene sequence, but appeared to be a member of the Eurasian lineage by the non-structural gene (NS) sequence suggesting that reassortment had occurred. A/eq/Switzerland/P112/07 was characterised as a member of the Eurasian lineage, the first time since 2005 that isolation of a virus from this lineage has been reported. Seven viruses from North America were classified as members of the Florida sublineage (clade 1), similar to A/eq/Wisconsin/03. In conclusion, a variety of antigenically distinct EIVs continue to circulate worldwide. Florida sublineage clade 1 viruses appear to predominate in North America, clade 2 viruses in Europe.     

Comparing needle and shoot phenology with xylem development on three conifer species in Italy

? Since the 1960s biologists have been trying to assess factors affecting cambial activity and latewood formation. A comparison of the phenology of the apical and lateral meristems could add new clues in order to understand mechanisms of the growth dynamic in conifers.

? This study compared needle and shoot growth with xylogenesis in Larix decidua, Pinus cembra and Picea abies during 2001 to verify if cambial resumption occurred after shoot and needle growth had begun, and if latewood initiation occurred when shoot lengthening was complete.

? In pine and spruce, needle and shoot lengthening was synchronous between mid-June and August while larches showed an early development of the needles at the end of May with a later shoot lengthening. Cell production lasted from mid-May to the beginning of August, with the first differentiating cells observed at the end of May, which indicated a earlier reactivation of the cambium with respect to needle and shoot growth.The first latewood cells began wall thickening between July and August, during the termination of needle and shoot lengthening.

? The hypothesis that cambial resumption occurs after shoot and needle elongation was rejected. The separation in time between apical growth and secondary wall thickening of latewood suggested the presence of an internal competition in resource allocation.

     

Antioxidant and antiproliferative activities of Spirulina and Chlorella water extracts

Liver fibrosis is a chronic liver disease that will further develop to cirrhosis if severe damage continues to form. A potential treatment for liver fibrosis is to inhibit activated hepatic stellate cell (HSC) proliferation and, subsequently, to induce HSC apoptosis. It has been reported that antioxidants are able to inhibit the proliferation of HSCs. In this study, the aqueous extract of spirulina was chosen as the source of antioxidant to investigate the inhibitory effect on the proliferation of HSC. The growth inhibitory effects of aqueous spirulina and chlorella extract on human liver cancer cells, HepG2, were also studied and compared in pairs. Results indicated that the total phenol content of spirulina was almost five times greater than that of chlorella (6.86 +/- 0.58 vs 1.44 +/- 0.04 mg tannic acid equivalent/g of algae powder, respectively). The antioxidant activity of spirulina determined by the ABTS*+ method was higher than chlorella (EC50: 72.44 +/- 0.24 micromol of trolox equivalent/g of spirulina extract vs 56.09 +/- 1.99 micromol of trolox equivalent/g of chlorella extract). Results of DPPH* assay also showed a similar trend as the ABTS*+ assay (EC50: 19.39 +/- 0.65 micromol of ascorbic acid equivalent/g of spirulina extract vs 14.04 +/- 1.06 micromol of ascorbic acid equivalent/g of chlorella extract). The aqueous extracts of these two algae both showed antiproliferative effects on HSC and HepG2, but spirulina was a stronger inhibitor than chlorella. Annexin-V staining showed that aqueous extract of spirulina induced apoptosis of HSC after 12 h of treatment. In addition, the aqueous extract of spirulina triggered a cell cycle arrest of HSC at the G2/M phase.     

The effects of dietary supplements of polyunsaturated fatty acid on pearl oyster,Pinctada margaritifera L., gonad composition and reproductive output

Black‐lip pearl oyster, Pinctada margaritifera broodstock was collected from the wild. Egg production, hatching rate and larval development were compared between oysters induced to spawn within 2 days after collection in the wild (T1), oysters fed a pure microalgae diet during 24 days before spawning (T2) and oysters fed the same microalgal diet in which 10% of the algae were replaced with 2 μm polyunsaturated fatty acid (PUFA)‐rich microspheres (T3). Administration of lipid microspheres resulted in larger sized eggs, a higher percentage of D‐larvae and larger sized 48‐h‐old larvae (P<0.05). The total and neutral lipid contents of the gonad increased after oysters were fed with microalgae only or with supplementary diet. The major neutral and polar fractions of saturated fatty acid (SFA) were 16C and 18C fatty acids, and not influenced by the diet (P>0.05). The gonads of oysters fed supplementary PUFA contained more docosahexaenoic acid (DHA) and less monounsaturated fatty acids. Higher level of DHA in gonads of T3 was associated with oogenesis and embryogenesis success. The n‐3/n‐6 ratio in the neutral lipid fraction provides a good indication of the spawning condition and predicting egg size and hatching rate.