Publications

4889

The spread of the reed planthopper Pentastiridius leporinus and its associated pathogens Candidatus Arsenophonus phytopathogenicus (ARSEPH) and Candidatus Phytoplasma solani (PHYPSO) increasingly threatens sugar beet production in Central Europe. Current surveillance systems rely mainly on vector monitoring, visual field assessments and molecular diagnostics of individual vectors and plants, all of which provide valuable but spatially limited information. This study evaluated sugar beet pulp samples generated during routine factory quality assessment as a large- scale surveillance resource for PHYPSO. During the 2025/26 campaign, 4887 pulp samples representing 1954 delivery field locations were collected, of which 1237 samples of individual field locations with consistent beet quality parameters were analyzed by direct PCR. PHYPSO was detected in 214 samples (17.3%). The molecular workflow was not subgroup-specific and did not include genotyping; therefore, PHYPSO-positive samples could not be assigned to individual 16SrXII subgroups. Comparison with visual field assessments showed that 85.7% of positive molecular detections originated from fields without visible symptoms. A carry-over experiment demonstrated that pathogen-derived DNA may persist across consecutive samples under worst-case conditions and must therefore be considered during data interpretation. In contrast, ARSEPH was detected by direct PCR in only six of 1237 samples (0.5%), all originating from the Euskirchen growing region. Comparison with qPCR, single-beet diagnostics and the carry-over experiment indicated insufficient sensitivity of the current ARSEPH direct-PCR assay; therefore, the observed detection rate should not be interpreted as true pathogen prevalence. Nevertheless, the observed spatial distribution patterns of monitoring sites indicate that industrial pulp sample monitoring substantially expands surveillance coverage beyond conventional symptom-based approaches and can help identify regions warranting further epidemiological investigation.

‘Candidatus Liberibacter solanacearum’ (Lso) haplotype D, transmitted by the carrot psyllid Bactericera trigonica, is a major threat to carrot production, particularly in the Mediterranean Basin and the Middle East. We investigated the impact of plant age at the time of inoculation on symptom development and yield under greenhouse and outdoor conditions. In greenhouse trials, 1- and 2-month-old plants were inoculated with psyllids harboring Lso. Both age groups showed reduced taproot weight compared with controls, but younger plants were more severely affected, exhibiting 90% yield reduction, while 2-month-old plants showed a 30% reduction. Witches’ broom symptoms appeared earlier in plants inoculated at 1 month (5 weeks postinoculation); however, within 2 weeks, plants inoculated at 2 months reached comparable levels of symptom incidence and severity. Additional greenhouse experiments using Lso-free psyllids confirmed that yield losses and disease symptoms were specifically due to Lso infection and not psyllid feeding. In outdoor trials, we tested the effects of inoculation at 7, 11, 15, and 19 weeks postsowing (wps) on disease incidence and yield. Significant yield losses occurred only in plants inoculated at 7 and 11 wps, with up to 45% reduction by harvest time. Symptoms began to appear at 20 wps in plants inoculated at 7 and 11 wps. At 23 wps, symptoms also appeared in plants inoculated at 15 wps, with a similar incidence to that observed in the earlier inoculation groups. Molecular detection of Lso at harvest (23 wps) confirmed that all plants inoculated at 19 wps or earlier were infected. These results demonstrate that carrots are highly susceptible to Lso-induced yield losses at the earlier stages of growth and highlight the importance of managing early season vector pressure to minimize economic damage.

“Candidatus Sarcina troglodytae” (“Ca. S. troglodytae”), a proposed bacterial species in the family Clostridiaceae, has been epidemiologically associated with a lethal disease in sanctuary chimpanzees in Sierra Leone, epizootic neurologic and gastroenteric syndrome (ENGS). As sarcinae are known to be difficult to culture, a lack of viable in vitro growth conditions has limited further characterization of this bacterium. Such studies are critical to elucidating the relationship between “Ca. S. troglodytae” and ENGS.  Here, we isolate a Gram-stain-positive, endospore-forming, coccus and anaerobic bacterial strain, designated JB3T, from brain tissue of a western chimpanzee (Pan troglodytes verus) using a reinforced clostridial medium. We describe the growth of strain JB3T at a pH range of 6 to 9 (optimum 6) at 37°C in an atmosphere composed of 5% H2, 5% CO2, and 90% N2 and demonstrate endospore production under alkaline conditions, suggesting the possibility of an environmental reservoir. We identify ethanol as the primary non-gaseous fermentation byproduct which we hypothesize to have relevance for ENGS. Antibiotic susceptibility testing shows susceptibility to common antimicrobial agents with notable resistance to ceftriaxone. By establishing in vitro conditions for the viable culture, propagation, and storage of “Ca. S. troglodytae”, this study lays the groundwork for future infection studies to test hypotheses about pathogenic mechanisms and prevention/treatment measures for ENGS.

Candidatus (Ca.) Allocryptoplasma is a newly identified, genetically diverse genus of vector-borne bacteria closely related to established blood-borne zoonotic pathogens, including Anaplasma, Ehrlichia, and Neoehrlichia species. Ca. Allocryptoplasma is a globally distributed pathogen detected across taxonomically diverse tick species and varied ecological niches. It maintains a complex enzootic circulation involving multiple vertebrate classes, including birds, small mammals, primates, and particularly reptiles, which serve as reservoir hosts. Methods: To characterize this emerging agent, multi-locus genetic DNA sequence analyses using the maximum likelihood method were performed on 16S rRNA, sucA, and rpoB genes amplified from the blood and vectors of marine iguanas and lizard species sampled across multiple islands in the Galápagos archipelago. Results: The compiled genomic data demonstrate that the Ca. Allocryptoplasma detected in the Galápagos lizards and marine iguanas represents a distinct lineage and a novel Ca. Allocryptoplasma species. Conclusions: Given its broad host plasticity, widespread geographic footprint, and vector-mediated transmission, the potential of Ca. Allocryptoplasma as a hidden, emerging zoonotic blood pathogen warrants intensive further evaluation.

Objective. To evaluate the effectiveness of eradicating HLB-symptomatic trees in reducing the spread of CLas. Materials and methods. The study was conducted from January to December 2019 in three phytosanitary epidemiological management areas (AMEFIs in Spanish) located in the municipalities of Güémez, Hidalgo, Llera de Canales, and Padilla, in the state of Tamaulipas, Mexico. In each AMEFI, a post-eradication sampling route was established around a tree confirmed to be symptomatic of CLas via q-PCR. Sampling sites were defined at distances of 6.7 km (S1), 13.4 km (S2), and 20.1 km (S3) from the eradicated tree, where leaf samples were randomly collected from asymptomatic trees to determine the presence or absence of CLas. The data were analyzed using chi-square tests to compare the frequency of positive and negative trees among AMEFIs and sampling sites. Results. The frequency of asymptomatic trees that were positive and negative for CLas differed significantly among the evaluated AMEFIs; at the site level, only AMEFI 3 showed significant differences. Conclusion. The eradication of trees with HLB symptoms was not sufficient to prevent the persistence of CLas, as asymptomatic infected trees were detected after the removal of symptomatic trees.