Serologic and molecular characterization of tickborne pathogens in lions (Panthera leo) from the Fasano Safari Park, Italy.

Lions (Panthera leo) are an endangered species threatened by illegal hunting, habitat loss, and infectious diseases. Little is known about the tick-borne pathogens that infect lions and could contribute to population declines. The objective of this study was to characterize Rickettsia spp., Anaplasma phagocytophilum, and Coxiella burnetii infections in 10 lions from the Fasano Safari Park in Italy by serology, polymerase chain reaction, and sequence analysis. Although animals did not show clinical signs of tick-borne diseases, evidence of infection with C. burnetii, spotted fever group Rickettsia sp., and A. phagocytophilum were found in 50%, 20%, and 10% of the lions, respectively. One of the lions tested positive for all three pathogens. This study is the first report of molecular evidence of infection with C. burnetii, Rickettsia sp., and A. phagocytophilum in lions and provides evidence that these felids become infected and serve as hosts for tick-transmitted bacteria.     

Validation of RT‐PCR Assays for Molecular Characterization of Porcine Teschoviruses and Enteroviruses

Porcine enteroviruses (PEVs) and teschoviruses (PTVs) are described as causative agents of neurological disorders, fertility disorders and dermal lesions of swine. Difficulties in the serological detection of these viruses may lead to a significant underestimation of infections with clinical symptoms. With the recent availability of genome sequence data for all the serotypes, molecular diagnosis is a possibility. The present study describes a new approach to molecular ‘serotyping’ of PTVs and PEV‐B viruses, involving the amplification and sequencing of a genomic fragment of the VP1 coding region. A molecular characterization of Italian entero‐teschovirus isolates was performed using a set of previously published and newly designed polymerase chain reaction primers. A total of 33 porcine isolates and 10 reference strains were analysed. Porcine enterovirus‐B samples were first diagnosed as positive for enterovirus by amplification of the 5′‐non‐translated region. Samples were then typed by amplification and sequencing of a portion of the VP1 coding region. Porcine enterovirus‐A and PTVs were detected by a published assay in the 5′‐NC region that allows them to be differentiated according to the size of amplification product, using the same set of primers. For serotype characterization of PTV, we evaluated four different regions: the N terminus of the capsid protein VP2, the region encoding for RNA‐dependent RNA polymerase, and the capsid VP1 and VP4 regions. The newly designed primers in the VP1 region was proved to be broad in range and suitable for serotype assessment and therefore constitute a useful diagnostic tool for molecular diagnosis of porcine teschovirus/enterovirus strains and for the study of molecular epidemiology and evolution of these viruses.     

Phytoremediation and Bio-physical Conditioning of Dredged Marine Sediments for Their Re-use in the Environment

A meso-scale pilot plant was set up to test the efficiency of a phytoremediation technique applied to slightly contaminated marine sediments dredged from the port of Livorno (central Italy). The technique applied involved the use of plants (a grass species, Paspalum vaginatum and a shrub species, Tamarix gallica) and earthworms (Eisenia foetida sp.) in order to recreate an active ecosystem in which plants, micro- and macroorganisms could interact with each other through the rhizosphere, for the adsorption, extraction and/or degradation of pollutants. This system enables plants to grow in a fine textured and salt medium highly unsuitable for plant species, and a physical, chemical and biological amelioration of the matrix was observed at the end of the experimentation (after 9 months). The sediment was bio-physically pre-conditioned by mixing it with calcareous material from excavating activities (5:1?v/v) and applying green compost. The presence of microorganisms in the rhizosphere and the earthworms produced a reduction by about 50% of the initial total petroleum hydrocarbon content. The use of a natural chelator with phyto-hormonal properties (humic substances) showed their capability of forming stable complexes, especially with Pb, and their contribution as an agronomic fertiliser. The combined effect of the P. vaginatum and T. gallica in the extraction of heavy metals from the sediment showed promisingly high percentages of metal effectively extracted in comparison with its available fraction (e.g. available Ni = 6.2%; Ni extracted = 15.7%). The technique proposed could be promising for an operational re-use of dredged marine sediments for agricultural purposes, a suitable final destination for the great majority of these contaminated materials from rivers and harbour docks.     

Correction of ADA-SCID by stem cell gene therapy combined with nonmyeloablative conditioning

Hematopoietic stem cell (HSC) gene therapy for adenosine deaminase (ADA)-deficient severe combined immunodeficiency (SCID) has shown limited clinical efficacy because of the small proportion of engrafted genetically corrected HSCs. We describe an improved protocol for gene transfer into HSCs associated with nonmyeloablative conditioning. This protocol was used in two patients for whom enzyme replacement therapy was not available, which allowed the effect of gene therapy alone to be evaluated. Sustained engraftment of engineered HSCs with differentiation into multiple lineages resulted in increased lymphocyte counts, improved immune functions (including antigen-specific responses), and lower toxic metabolites. Both patients are currently at home and clinically well, with normal growth and development. These results indicate the safety and efficacy of HSC gene therapy combined with nonmyeloablative conditioning for the treatment of SCID.     

Soil Bioremediation: Combination of Earthworms and Compost for the Ecological Remediation of a Hydrocarbon Polluted Soil

A three-dimensional (3D) off-line chemical transport model (CTM) has been used to study an Arctic ozone minihole event in early December 2002, when the meteorological conditon were severely colder than in recent years. The minimum temperatures were below 188 K on 6 December 2002 and remained below 195 K for the entire month. Such temperatures are low enough for the formation of PSCs (polar stratospheric clouds) which were observed by a visible/near infrared solar occultation instrument by the Polar Ozone and Aerosol Measurement (POAM) III experiment. The SLIMCAT model compares well with the depth and evolution of the ozone distribution as observed by the second European Remote Sensing Satellite (ERS-2) Global Ozone Monitoring Experiment (GOME). The combined effect of a very strong and large intrusion of sub-tropical air, and uplifting of air masses of low ozone concentrations from the troposphere plays important role in the fomation of the ozone minihole. The modelled chemical ozone loss rate was relatively fast in the lower stratosphere in the region of frequent PSCs and higher levels of ClO and BrO. The observed PSCs in the vicinity of the minihole supports the model-predicted location of PSCs and chlorine activation. However, ozone miniholes have been attributed almost entirely to dynamical effects due to its short duration.     

Clostridium perfringens toxin-types in lambs and kids affected with gastroenteric pathologies in Italy

A study was carried out in the South of Italy to assess the role of clostridia in neonatal diseases of lambs and kids. Eighty-seven lambs and 15 kids belonging to 25 flocks were examined and Clostridium perfringens was the microorganism most commonly identified. C. perfringens isolates were analysed by polymerase chain reaction (PCR), in order to determine the prevalence of the genes cpa, cpb, cpb2, etx, iap and cpe. The most prevalent toxin-type of C. perfringens was found to be type A found in 84% of the cases with clostridial enterotoxaemia. No C. perfringens type B, C or E were found. C. perfringens type D was isolated in 16% of the cases. About 24% of the isolates were cpb2 positive. The prevalence of cpb2 across the different C. perfringens types varied. The beta(2)-toxin gene cpb2 was detected in 4/21 (19%) type A isolates, in 1/2 type D isolates, and in 1/2 type DE (cpe-carrying type D) isolates. The high rate of positivity to cpb2 among the isolates suggests that a vaccine based on the beta(2)-toxin, should be included in the vaccination schedule of the animals to confer adequate protection and to prevent the disease.     

A century of Alzheimer's disease

One hundred years ago a small group of psychiatrists described the abnormal protein deposits in the brain that define the most common neurodegenerative diseases. Over the past 25 years, it has become clear that the proteins forming the deposits are central to the disease process. Amyloid-beta and tau make up the plaques and tangles of Alzheimer's disease, where these normally soluble proteins assemble into amyloid-like filaments. Tau inclusions are also found in a number of related disorders. Genetic studies have shown that dysfunction of amyloid-beta or tau is sufficient to cause dementia. The ongoing molecular dissection of the neurodegenerative pathways is expected to lead to a true understanding of disease pathogenesis.     

Genomic characterization of H1N2 swine influenza viruses in Italy

Three subtypes (H1N1, H1N2, and H3N2) are currently diffused worldwide in pigs. The H1N2 subtype was detected for the first time in Italian pigs in 1998. To investigate the genetic characteristics and the molecular evolution of this subtype in Italy, we conducted a phylogenetic analysis of whole genome sequences of 26 strains isolated from 1998 to 2010. Phylogenetic analysis of HA and NA genes showed differences between the older (1998-2003) and the more recent strains (2003-2010). The older isolates were closely related to the established European H1N2 lineage, whereas the more recent isolates possessed a different NA deriving from recent human H3N2 viruses. Two other reassortant H1N2 strains have been detected: A/sw/It/22530/02 has the HA gene that is closely related to H1N1 viruses; A/sw/It/58769/10 is an uncommon strain with an HA that is closely related to H1N1 and an NA similar to H3N2 SIVs. Amino acid analysis revealed interesting features: a deletion of two amino acids (146-147) in the HA gene of the recent isolates and two strains isolated in 1998; the presence of the uncommon aa change (N66S), in the PB1-F2 protein in strains isolated from 2009 to 2010, which is said to have contributed to the increased virulence. These results demonstrate the importance of pigs as mixing vessels for animal and human influenza and show the presence and establishment of reassortant strains involving human viruses in pigs in Italy. These findings also highlighted different genomic characteristics of the NA gene the recent Italian strains compared to circulating European viruses.     

Balancing resource protection and fishing activity: The case of the European hake in the northern Iberian Peninsula

One of the fundamental aims of the Ecosystem Approach to Fisheries Management is to ensure the long‐term sustainability of the fishery by protecting key life‐cycle habitats, such as recruitment areas. In this study, we apply a hurdle Bayesian hierarchical spatio‐temporal model that describes the abundance of European hake (Merluccius merluccius) recruits in the northern continental shelf of the Iberian Peninsula. Our findings clearly show four persistent nurseries, the main one being located along the continental shelf of the Artabrian gulf (off La Coruña). The preferential habitats identified for the hake recruits are areas within a bathymetric range of 120–200 m, with 15–16°C of Sea Surface Temperature, a Chl‐a concentration of 0.8–1.2 mg/m³ and low values of seabed rugosity (unconsolidated substrates). Searching for a compromise with fisheries, we also assess the degree of overlap of the main nursery areas with two fisheries footprints, a local one using Vessel Monitoring System data of trawl fishery of Marín, and a global one using Automatic Identification System data of different type of trawlers (bottom otter, beam and midwater trawls). The two fisheries footprints present different degree of overlapping in distinct areas, highlight the need of specific fleet‐adapted management rules in order to protect juvenile stages. We discuss that understanding the distributional patterns associated with key life stages, such as recruitment, and their interaction with fishing activities, is essential for applying appropriate spatial management measures and improving fishery sustainability.     

1H‐NMR metabolomic profiling of the crayfish Astacus leptodactylus subjected to polyphenol‐enriched diets

¹H‐NMR analysis of the hepatopancreas, muscle and haemolymph of Astacus leptodactylus after feeding with polyphenol‐enriched diet is reported. ¹H‐NMR spectra of lipophilic extracts showed the presence of cholesterol, fatty acid residues, phospholipids and triglycerides. ¹H‐NMR spectra of aqueous extracts identified 35 metabolites in the hepatopancreas, 31 in the muscle and 22 in the haemolymph. A total of 20 metabolites (amino acids and their derivatives) were present in the hepatopancreas, the muscle and the haemolymph. A total of 10 metabolites were present in both the hepatopancreas and the muscle (five amino acids, 2‐hydroxybutyrate, choline, myo‐inositol, glycogen and uracil). 2‐Hydroxyisobutyrate and creatine were present in both the hepatopancreas and the haemolymph. Phosphorylethanolamine, phosphocholine and fumarate were present only in the hepatopancreas and isoleucine only in the muscle. Statistical analysis showed that the percentage of weight gain was statistically higher in polyphenol‐enriched diet groups compared to the control and that polyphenols had a stimulating effect on the general metabolism. No stress‐related metabolites were higher in crayfish fed with polyphenol‐enriched diet. Conversely, phosphatidylcholine, cholesterol and DHA, linked to resistance to environmental stress and diseases, were higher compared to the control diet. This study indicates that ¹H‐NMR is a useful tool to study the metabolomics in relation to diet differences.