Geminivirus DNA replication and cell cycle interactions

The Geminiviridae family includes a large number of viruses that infect plants and have a unique geminate virion particle, a single-stranded genome of approximately 2.6-3.0 kb, and replicate through a rolling-circle mechanism. Since they encode for just a few proteins (4-6 depending on the members that belong to four different genera), a rich variety of interactions has evolved between viral proteins and host factors to develop the virus replicative cycle. Among them, we have been particularly interested so far: (i). in the interference with cell cycle regulatory proteins of the retinoblastoma-related (RBR)/E2F pathway and (ii). in the interaction with host DNA replication factors necessary for the assembly of a functional replication complex at the viral origin of DNA replication during the rolling-circle stage. Yeast two-hybrid assays revealed that wheat dwarf virus RepA protein, but nor Rep protein, interacts with plant RBR protein. Interestingly, deletion of the C-terminal domain of Rep confers the truncated protein the ability to interact with RBR, suggesting that this domain may hinder the LXCXE RBR-binding motif. Secondary structure predictions support such a possibility.     

Diagnosis and risk factors of Aelurostrongylus abstrusus (Nematoda, Strongylida) infection in cats from Italy

Aelurostrongylus abstrusus (Nematoda, Strongylida) in cats and associated risk factors have been investigated by classical coprology in central (site A) and southern (site B) Italy, i.e. by floatations with sugar and zinc sulphate solutions and a Baermann technique. Overall, 227 individual stool samples (i.e. 162 from site A and 65 from site B) were processed and risk factors were related to infection by binary logistic multiple-regression models. The presence of A. abstrusus was used as the dependent variable and the epidemiological data as independent variables. Twenty-eight (17.3%) and 12 (18.5%) cats were positive for A. abstrusus in sites A and B, respectively, both using the Baermann and flotation with zinc sulphate solution methods. Six and three (from site A and B, respectively) of the infected cats were negative for A. abstrusus when faeces were processed with sugar solution. Stray and free-ranging (P=0.357), young cats (P=0.008) and the presence of respiratory symptoms (P=0.000) were risk factors for A. abstrusus infection. The results suggest that A. abstrusus is spread in Italy. Furthermore, these results possibly include infection in the differential diagnosis of feline respiratory diseases and offer a correct diagnostic approach by using either a Baermann method or flotation with zinc sulphate solution in suspected cases.     

Contrast‐enhanced ultrasound complements two‐dimensional ultrasonography in diagnosing gallbladder diseases in dogs

Gall‐bladder diseases are common in dogs and two‐dimensional ultrasonography is a current standard method for diagnosis and treatment planning. However, findings from this modality can be nonspecific. The aim of this retrospective, case series study was to describe conventional and contrast‐enhanced ultrasound (using SonoVue^®) findings in a group of dogs with histologically confirmed gall bladder disease. A total of 65 dogs were included. Branchlike, heterogeneous, and homogeneous contrast enhancement of echogenic intraluminal mass‐forming lesions was a contrast‐enhanced ultrasound characteristic of polypoid lesions due to cystic mucosal hyperplasia of the gallbladder and/or tumor, which had different wash‐in and washout characteristics. In dogs with mobile or immobile biliary sludge or mucocele, the echogenic intraluminal masses remained unenhanced. A double rim mark or enhancement defect in the gallbladder wall was a characteristic of edema or necrosis/rupture of the wall, respectively. Conventional ultrasonography correctly identified biliary sludge or mucocele in 36/37 dogs, cholecystitis/edema in 44/47 dogs, necrosis/rupture in 19/25 dogs, and gallbladder neoplasia in three of three dogs with these pathologies. It falsely identified biliary sludge or mucocele in eight of 28 dogs, cholecystitis/edema in three of 15 dogs, necrosis/rupture in 13/37 dogs, and gall‐bladder neoplasia in 20/59 dogs that did not have these pathologies. Contrast‐enhanced ultrasound correctly identified cholecystitis/edema in 42/47 dogs, but falsely identified cholecystitis/edema in three of 18 dogs. It correctly identified necrosis/rupture, benign polypoid lesions, and gallbladder neoplasia in all dogs with no false‐positive results. Findings supported contrast‐enhanced ultrasound as a complement to conventional ultrasonography for dogs with suspected gallbladder pathologies such as edema, necrosis, and rupture.     

Efficacy of a combination of 10% imidacloprid/50% permethrin for the prevention of leishmaniasis in kennelled dogs in an endemic area

The efficacy of imidacloprid 10%and permethrin 50% (Advantix; Bayer AG, Germany) in a spot-on formulation was evaluated in the field as a control measure to prevent canine leishmaniasis (CanL) in dogs in an endemic area of southern Italy. In February 2005, out of 845 dogs initially tested for CanL, 631 dogs which tested negative (315 from a kennel in Bari (KB) and 316 from a kennel in Ginosa (KG)) in a serological and a parasitological examination were allocated to one of three groups: Group A-treated with imidacloprid 10% and permethrin 50% once a month; Group B-treated every 2 weeks; and Group C-untreated control animals. All the dogs were examined serologically and parasitologically for CanL prior to the start of the study, in November 2005 (end of the sandfly season) and in March 2006 (end of the study). An initial CanL seroprevalence of 24.7% (209 dogs) was detected in KB and KG. In KB Leishmania infection, inferred by positivity in at least one of the three tests performed at the interim or final follow-up, was found in one animal from Group A and in nine from Group C. No positive animals were detected in Group B, thus giving a final protection efficacy of 88.9% in Group A and 100% in Group B. In KG Leishmania infection was identified in one animal from Groups A and B, respectively, and 11 from Group C (protection efficacy of 90.36% in Group A and 90.73% in Group B). The incidence density rates (IDRs) of infection in both Groups A and B at each kennel were statistically significantly lower than that registered in Group C (KB p<0.05 and KG p<0.01). The results clearly show that a combination of imidacloprid 10% and permethrin 50%, by virtue of its repellent activity against sandflies, is effective under both application regimes in preventing CanL in the field in endemic areas.     

Occurrence of anthelmintic resistant equine cyathostome populations in central and southern Italy

In the present survey, 276 horses bred on 16 farms located in central and southern Italy were investigated for the presence of drug resistant cyathostomes by a Fecal Egg Count Reduction Test (FECRT). Sixteen to 20 animals were selected on each farm and randomly assigned to one of four equally sized treatment groups. Groups were treated with fenbendazole, pyrantel pamoate, ivermectin or moxidectin. Resistance to fenbendazole was declared on six farms (37.5%) and suspected in two farms (12.5%), with FECR values ranging from 41% to 88.3%. Resistance to pyrantel was found in two farms (12.5%) and was suspected in one case (6.2%), with FECR values ranging from 43% to 85.4%. Macrocyclic lactones remained effective on all farms. Only cyathostome third stage larvae (L3) were found in fecal cultures after treatment. This paper reports the first wide survey conducted in Italy for anthelmintic resistance in equine cyathostomes. The results indicate that multiple drug resistant equine cyathostomes are present in the central and southern regions of Italy. These data call for a geographically and numerically broader investigation of horse farms in all regions and for the development and implementation -among veterinarians, owners and managers of a plan to reduce the expansion of these anthelmintic resistant populations and control these important parasites.     

Fish and seafood traceability based on AFLP markers: Elaboration of a species database

Several sociological, health and conservation arguments request a correct labelling of seafood products. Nowadays, molecular genetics is a useful tool for food chain traceability, particularly in regards to species identification. Among the variety of PCR-based molecular markers, AFLPs (Amplified Fragment Length Polymorphisms) have recently been used to investigate genomes of different complexities. This paper assesses the potential use of the AFLP technology to determine fish and seafood species in processed commercial products and domestic stocks. In particular a species database of fish, molluscs and crustaceans has been created with the aim to identify species of origin of seafood products by previously defined AFLP patterns. Different EcoRI and TaqI primer combinations were selected from 20 screened combinations in relation to the total number of detected fragments and polymorphic ones. Most informative combinations were E32/T32, E32/T33, E33/T33, E33/T37, E33/T38, E40/T33, E40/T37, E42/T32, E42/T37. The comparison of informative markers between unknown frozen or fresh products and reference samples has enabled the accurate identification of 32 different species. The taxonomic characterization has been performed either at the species or at the population level depending on the number of available individuals. AFLP variation at the population level is particularly helpful for the stock traceability of domestic strains. Size homoplasy was also investigated in one species to assess the rate of non-homologous comigrating fragments and to detect additional polymorphic markers to be used in stock identification. Results of Band Sharing Index (BSI) and percentage of polymorphic fragments are presented and are discussed in relation to the wide applicability of AFLPs both for fish and seafood safety and authenticity testing in such fields as food traceability and restocking management. The database, available upon request at nonnis@biol.unipr.it, will be continuously updated.     

Environmental influences on the recruitment dynamics of juvenile European eels,Anguilla anguilla,in a small estuary of the Tyrrhenian Sea,Sardinia, Italy

1. The European eel, Anguilla anguilla, is a catadromous and migratory species of commercial importance. Its complex life cycle results in its exposure to many risk factors, which have resulted in stock declines across all life stages since the 1970s.
2. The temporal recruitment dynamics of juvenile eels (glass eels and elvers) were investigated in a small Mediterranean estuary (Sardinia, Italy). The composition of the population and the monthly and seasonal variations in the abundances of juvenile eels was assessed over 78 sampling events (from February 2017 to February 2018). Furthermore, the effects of abiotic variables on the abundances of glass eels and elvers were investigated using generalized additive models (GAMs).
3. Glass eels had the greatest abundance during the winter months, whereas elvers had the greatest abundance during spring. Modelling revealed that the abundance of glass eels was mostly explained by the combined effects of water temperature (12.3–14.5 °C), tidal coefficient (40–110 cm), moon phase, season, and river mouth condition, whereas the abundance of elvers was associated with water temperature (14–21 °C), dissolved oxygen content (>7 mg/L), and season. These results suggest that the annual recruitment of juvenile eels occurs throughout the year, with clear seasonal migration dynamics.
4. The use of multiple statistical approaches allowed us to identify the importance of several environmental variables in regulating the recruitment dynamics, providing useful information for conserving eel stocks through the restoration of the natural flow regime and the connectivity between freshwater habitats and the sea.

     

Pb adsorption by soils

Pb adsorption for 12 soils from Tuscany was studied. The data fitted the Langmuir and the Freundlich isotherms over a large range of concentrations. Results showed that organic matter and clay content were responsible for adsorption maxima. The effect of Mn oxides, explained independently of organic matter and clay, was negligible. The adsorption maxima were generally found to be greater than CEC; the possible mechanisms are discussed.     

Assessment of apple (Malus × domestica Borkh.) fruit texture by a combined acoustic-mechanical profiling strategy

Texture of apple fruit originates from anatomic traits related to cell wall architecture and is one of its most important quality characteristics, thus there is the desire to better understand the different factors which contribute to apple texture. Here we present a novel approach based on the simultaneous profiling of the mechanical and acoustic response of the flesh tissue to compression, using a texture analyzer coupled with an acoustic device. The methodology was applied to a 86 different apple cultivars, measured after two months postharvest cold storage and characterised by 16 acoustic and mechanical parameters. Statistical treatment of the data with principal component analysis (PCA) allowed for the identification of three groups of variables, the mechanical ones being clearly distinguished from the acoustic ones. Moreover, the distribution of the apple cultivars in the multivariate PCA plot allowed characterisation of the cultivars according to their textural performance. Each cultivar was analyzed also with non-destructive vis/NIR spectroscopy in order to determine impartially the ripening stage. Sensory evaluation by panellists was performed on a selected group of cultivars and sensory data correlated with the acoustic-mechanical data. The results demonstrate the good performance of our combined acoustic-mechanical strategy in measuring apple crispness as it is perceived by human senses.