Integrative diagnosis, biological observations, and histopathology of the fig cyst nematode Heterodera fici Kirjanova (1954) associated with Ficus carica L. in southern Italy

Morpho-biological notes and histopathology, based on LM and SEM observations, of the fig cyst nematode Heterodera fici isolated from Ficus carica roots, collected in home and public gardens of Apulia region, southern Italy, are described and illustrated. Seventy-five localities throughout the Apulia region were sampled and one-quarter of the sampled localities had fig roots infested with H. fici, with population densities ranging from 44 to 180 cysts/100 ml of soil. All attempts to detect H. fici on ornamental Ficus spp. as well as on imported bonsai in Italy were unsuccessful. Morphometric characters of the Italian population conform to those of the type and re-description populations reported for H. fici. Molecular analysis using ITS, D2–D3 expansion domains of the 28S rRNA, and the partial 18S rRNA sequences of H. fici newly obtained in this study matched well with the corresponding sequences of H. fici present in the GenBank database. Phylogenetic trees confirmed and supported the grouping of H. fici in the Humuli group. Heterodera fici completes its embryogenic development in 14–16 days at 25 °C. Post-invasion development and maturity in the roots of F. carica seedlings is completed in 64–68 days at 25–28 °C with juveniles and adults showing different parasitic habits, being endoparasitic and semi-endoparasitic respectively. The establishment of permanent feeding sites that consist of the formation of large syncytia causes anatomical modification of vascular elements and general disorder in the root stelar structures. Syncytia structures associated with mature females showed different degrees of vacuolisation, numbers of syncytial cells, and contained nuclei and nucleoli which were constantly hypertrophied

A polyphasic approach with molecular phylogeny for the characterization of Anisakis pegreffii (Anisakidae: Nematoda) in fishes from Adriatic Sea

Abstract In this study we investigated the morphometric and molecular characterization of a liver encapsulated third-stage larval population of Anisakis spp. infecting Merluccius merluccius and Lophius piscatorius caught in the Adriatic Sea waters (southern Italy). A polyphasic approach based on PCR-RFLP profiles of the ITS region, mitochondrial COI (cytochrome c oxidase subunit 1), sequencing and molecular phylogeny of ITS and mitochondrial COI was used to identify Anisakis larvae collected from fish samples. PCR-RFLP analysis showed three banding pattern corresponding to the peculiar pattern of A. pegreffii. Sequence data from ribosomal ITS and mitochondrial COI were analysed by Neighbour Joining, Minimum Evolution and Maximum Parsimony methods to evaluate the phylogenetic relationships among A. simplex sensu lato. The phylogenetic trees obtained for both ITS and COI revealed the existence of three distinct clades for A. simplex sensu sricto, A. simplex C and A. pegreffii and the sequences obtained in this study clearly clustered together with A. pegreffii sequences present in the database. Histopathological observations of anisakid nematode specimens detected on the liver surface of M. merluccius are illustrated. Encapsulated specimens of the L3 stage of the nematode were similar in size and morphometry to those found into the peritoneal cavity. Anisakis larvae encapsulated on the liver surface within dense and pearl coloured envelops caused host hepatic tissue necrosis, large cavities and oedematous liver spots to the host

Parasitism effects on white clover by root-knot and cyst nematodes and molecular separation of Heterodera daverti from H. trifolii

This research carried out an accurate identification of the root-knot and cyst-forming nematode species parasitizing white clover at the Laceno Lake area in Southern Italy. Two species, the root-knot nematode Meloidogyne hapla and the cyst nematode Heterodera daverti were identified by integrative taxonomic approaches (classical, isozyme pattern, and D2-D3 expansion segments of 28S rRNA, ITS rRNA and coxI of mtDNA gene sequences) and found parasitizing white clover roots. These nematodes were detected in stunted plants with a reduced number of rhizobium nodules and the host suitability was confirmed by the high nematode population densities ranging from 53 to 2350 eggs and J2s per g of fresh roots for M. hapla, and 1.36 eggs and J2s/cm3 of soil for H. daverti; and cyst nematode females were also detected on the roots of clover. The studies on the host-parasite relationships of nematode-feeding sites in white clover roots infected by these nematodes showed a high susceptible response. Meloidogyne hapla and H. daverti infections were also observed on nitrogen-fixing root nodules of white clover, where well established feeding sites allowed active nematode reproduction. Histological examination of nitrogen-fixing root nodule tissues revealed that the nematodes established their permanent feeding sites in the vascular bundles of nodules which appeared enlarged deformed and disorganised by the expansion of nematode feeding cells (giant cells and syncytium) and hyperplasia of the nodule cortex. Additionally, coxI of mtDNA gene is an efficient barcoding sequence for discriminating the identification of H. daverti from H. trifolii.The present work has been done in the frame of the Apulia Region Programme: Misura 214/4 sub azione a “Progetti integrati per la Biodiversità” PSR 2007–2013, ASSE II “Miglioramento dell’ ambiente e dello Spazio Rurale” SaVeGraINPuglia.Peer reviewe

Cranial Trepanation: Case Studies between the IV Century BC and VI Century in Southern Italy

Cranial trepanation is among the oldest head surgical procedures, creating a hole through the skull. This procedure is the surgical removal of a portion of the skull as a medical treatment to cure a specific disease or as a religious or magical ritual. Cranial trepanation has been reported from very early times and widespread all over the world. This work is based on data from six trepanned skulls from the Hellenistic Age (IV century BC) to the Late Antiquity (VI century). The samples come from Puglia (Southern Italy), from the northern (the peninsula of Gargano) to the central area: three cases come from Vieste (Foggia), Ischitella (Foggia) and Alberona (Foggia), the others three from Gravina in Puglia (Bari) and Canosa (Barletta, Andria, Trani). All skulls have been evaluated via an anthropological, radiological and odontological approach. This study indicates the value of a methodology, which encompasses anthropology, radiology and imaging, and archeo-forensic investigations, highlighting the need for multidisciplinary teamwork in any assessment of human remains. The study supports the hypothesis of the presence of a long tradition of medical centres in the Mediterranean basin area

CRANIAL TREPANATION IN THE APULIAN REGION OF ITALY BETWEEN THE IV CENTURY BC AND VI CENTURY AC: CASE STUDIES

Cranial trepanation is among the oldest head surgical procedures, creating a hole through the skull. This procedure is the surgical removal of a portion of the skull as a medical treatment to cure a specific disease or as a religious or magical ritual. Cranial trepanation has been reported from very early times and widespread all over the world. This work is based on data from six trepanned skulls from the Hellenistic Age (IV century BC) to the Late Antiquity (VI century). The samples come from Puglia (Southern Italy), from the northern (the peninsula of Gargano) to the central area: three cases come from Vieste (Foggia), Ischitella (Foggia) and Alberona (Foggia), the others three from Gravina in Puglia (Bari) and Canosa (Barletta, Andria, Trani). All skulls have been evaluated via an anthropological, radiological and odontological approach. This study indicates the value of a methodology, which encompasses anthropology, radiology and imaging, and archeo-forensic investigations, highlighting the need for multidisciplinary teamwork in any assessment of human remains. The study supports the hypothesis of the presence of a long tradition of medical centres in the Mediterranean basin area