First Report of the Root-Knot Nematode, Meloidogyne hispanica, Infecting Sunflower in Greece.

Severe plant stunting, chlorosis, and extensive root galling were observed on sunflower (Helianthus annus Pioneer Hi-bred PR64LE19, Dupont) in a commercial field at Agios Athanasios, Drama Province, northeastern Greece at the end of May 2013. Disease symptoms were observed about 1.5 months after planting, and were distributed in patches that covered approximately 2% of the whole cultivated area. Examination of the soil and root samples from selected infected plants revealed the presence of abundant root-knot nematodes. Juveniles, males, and females were extracted by sieving, decanting, and root dissection for identification using morphological traits. Nematode population densities ranging from 100 to 150 J2s per 100 cm3 of soil, and 150 to 3,000 eggs per g of fresh sunflower roots were observed. Identification was confirmed by perineal patterns of females and by sequencing of the D2-D3 expansion segments of 28S ribosomal RNA gene (1,3,4). All identification methods were consistent with typical Meloidogyne hispanica. Morphology of perineal patterns of females and measurements of the second-stage juveniles (J2s) matched those of the original description of M. hispanica (3). Alignment indicated that the D2-D3 sequence (GenBank Accession No. KF501128) was 99% homologous to other sequences of M. hispanica deposited in GenBank from Brazil, Portugal, and Spain (EU443606, EU443608, and GQ375158, respectively), differing in only one nucleotide. Phylogenetic analyses using maximum likelihood of this sequence placed the Meloidogyne sp. in a highly supported (100%) clade that included all M. hispanica sequences available from the GenBank database (4). Root-knot nematodes in general have been reported to cause economic losses in sunflower in Europe (2), but there are no reports of M. hispanica. M. hispanica was first found in Seville Province, southern Spain, infecting rootstocks of Prunus spp. (3). Its distribution has been confirmed worldwide on different agricultural crops. However, to our knowledge, this is the first report of M. hispanica infecting sunflower in Europe and the first report of this species on any crop for Greece. The identification of M. hispanica in sunflower is relevant because it may represent a threat for sunflower production in Greece. Research to develop sunflower varieties resistant to root-knot nematodes should now also consider M. hispanica along with other species of Meloidogyne. References: (1) K. R. Barker. Page 19 in: An Advanced Treatise on Meloidogyne. Vol. II, Methodology. K. R. Barker et al., eds. North Carolina State University Graphics, Raleigh, NC, 1985. (2) M. Di Vito et al. Nematol. Mediterr. 24:109, 1996. (3) H. Hirschmann. J. Nematol. 18:520, 1986. (4) B. B. Landa et al. Plant Dis. 92:1104, 2008.

Molecular and Morphological Characterization of the Corn Cyst Nematode, Heterodera zeae, from Greece.

The corn cyst nematode Heterodera zeae was detected in soil from an organic maize field in northern Greece. In greenhouse studies, reproduction of H. zeae was detected on maize plants (Zeae mays) using soil high in organic matter; the field was under winter fallow at the time of sampling. Maize plants were grown in a greenhouse with soil from the affected field used as inoculum. Females appeared after six weeks incubation, and abundant cysts were present after 12 weeks. Morphological and molecular diagnosis confirmed the presence of H. zeae in the field. Cysts were identified on the basis of cyst shape and characteristics of the cyst terminal cone, including nature of fenestration, presence of bullae, cyst wall pattern, and fenestral diameter. Second-stage juveniles were identified by body and stylet length, the shape of stylet knobs, shape and length of the tail and hyaline tail terminus, and by the number of lateral lines. Molecular analysis included amplification of the ribosomal internal transcribed spacer regions (ITS 1&2 rDNA) 28S large ribosomal subunit (LSU) D2-D3 expansion segment, and partial 18S small ribosomal subunit (SSU). Restriction fragment length polymorphism (RFLP) of ITS rDNA exhibited several unique enzyme patterns that may be diagnostically useful for H. zeae. These findings are in agreement with prior analysis of H. zeae populations from the U.S. and India. Phylogenetic relationships inferred from ITS rDNA are congruent with previous analyses that placed H. zeae in a clade with H. turcomanica, H. salixophila and species of the Humuli group. Phylogenetic trees based upon heat shock protein (Hsp90) coding sequence were in general agreement with a prior study using the same marker. This study represents the first record of H. zeae in Greece and the second report of this nematode in Europe.

Symptomatic hypercalcemia as presenting manifestation of lymphoma in a patient with acquired immunodeficiency syndrome.

Hypercalcemia can be an uncommon manifestation of various infections and malignancies in HIV-positive patients. A case of severe symptomatic hypercalcemia as the first sign of non-Hodgkin's lymphoma in a patient found later to be HIV positive is presented. Differential diagnosis of calcium disorders should include test for HIV and underlying conditions including lymphoma in the diagnostic work up.

Longidorus cretensis n. sp. (Nematoda: Longidoridae) from a vineyard infected with a foliar 'yellow mosaic' on Crete, Greece.

A description is provided of Longidorus cretensis n. sp., a bisexual species associated with grapevines in Crete, Greece. The species is characterised by a body length of 6.3-8.7 mm, head flattened anteriorly and continuous with the body profile, amphids pocket-shaped and not bilobed, a posteriorly situated guide-ring, posteriorly situated oesophageal glands, and a tail bluntly rounded to hemispherical. Xiphinema index was present throughout the vineyard, but L. cretensis n. sp. occurred only in an area where the grapevine foliage exhibited a 'yellow mosaic' disease, and in association with X. pachtaicum from the rhizosphere of an olive tree growing adjacent to the vineyard. Nematode-transmitted virus was not recovered from weed species growing in the vicinity of the olive tree, or by bait-testing soil collected from the rhizosphere of the tree. Population densities of L. cretensis n. sp. were similar under olive and grapevine, but twice as many adult specimens were recovered from soil samples collected in the vineyard. Three quarters of L. cretensis specimens were present at 21-40 cm depth under olive, whereas a similar proportion of the population was equally distributed under grapevine at 21-40 cm and 41-60 cm depths.

Neutropenia induced by paraquat poisoning.

Paraquat is a synthetic, nonselective, contact herbicide, which causes injury to tissues following contact. When ingested in sufficient amounts, paraquat has life-threatening effects on the gastrointestinal tract, lungs, kidney, liver, heart and other organs. This is a case report of a 15-year-old girl from an agricultural area who was admitted to hospital for treatment. She presented with discomfort, nausea and vomiting. She had drunk a small wine glass full of paraquat. After 18 days in the hospital, the girl developed neutropenia. This is the first reported case of paraquat poisoning-induced neutropenia.

Evidence for a Dosage Effect of the Mi gene on Partially Virulent Isolates of Meloidogyne javanica.

The reproduction of single egg-mass isolates of Meloidogyne javanica from Crete that differed in virulence were compared on tomato (Lycopersicon esculentum) genotypes homozygous or heterozygous for the Mi gene. The reproduction of three isolates with partial virulence was much greater on tomato genotypes heterozygous for the Mi gene (cultivars Scala, Bermuda, and 7353) than on two homozygous genotypes (F8 inbred lines derived from Scala). The reproduction of a highly virulent isolate on the homozygous and heterozygous genotypes was similar to that on a susceptible cultivar. These results pose questions regarding the nature of partial virulence and indicate a quantitative effect of the Mi gene in relation to such virulence.