A 314-bp SINE insertion in the ZNF2 promoter region may act as a repressor related to regulation of fat deposition in pigs

Retrotransposons, a type of DNA fragment that can mobilize itself on genome, can generate genetic variations and develop for molecular markers based on the insertion polymorphism. Zinc finger proteins (ZNFs) are among the most abundant proteins in eukaryotic animals, and their functions are extraordinarily diverse and particularly important in gene regulation. In the current study, bioinformatic prediction was performed to screen for retrotransposon insertion polymorphisms (RIPs) in six ZNF genes (ZNF2, ZNF3, ZNF7, ZNF8, ZNF10 and ZNF12). Six RIPs in these ZNFs, including one short interspersed nuclear element (SINE) RIP in intron 1 and one long interspersed nuclear element 1 (L1) RIP in intron 3 of ZNF2, one SINE RIP in 5′ flanking region and one SINE RIP in intron 2 of ZNF3, one SINE RIP in 3′ UTR of ZNF7 and one L1 RIP in intron 2 of ZNF12, were discovered and their presence was confirmed by PCR. The impact of the SINE RIP in the first intron of ZNF2, which is close to the core promoter of ZNF2, on the gene activity was investigated by dual-luciferase assay in three cell lines. Our results showed that the SINE insertion in the intron 1 of ZNF2 repressed the core promoter activity extremely significantly (P<0.01) in cervical cancer cells and porcine primary embryonic fibroblasts (HeLa and PEF), thus SINE may act as a repressor. This SINE RIP also significantly (P<0.05) affected the corrected back fat thickness in Yorkshire pigs. The corrected back fat thickness of individuals with SINE insertion in the first intron of ZNF2 was significantly (P<0.05) higher than that of individuals without SINE insertion. In summary, our data suggested that RIPs play important roles in the genetic variations of these ZNF genes and SINE RIP in the intron 1 of ZNF2 may provide a useful molecular marker for the screening of fat deposition in the pig breeding.     

Post-infection activity of fungicides against <Emphasis Type="Italic">Phytophthora infestans</Emphasis> on tomato (<Emphasis Type="Italic">Solanum lycopersicum</Emphasis> L.)

Late blight caused by Phytophthora infestans is the most devastating disease of tomato (Solanum lycopersicum L.) and causes important economically losses if not properly controlled. Control is achieved mainly by preventive fungicide applications. However, even if curative applications are discouraged because they increase the risk of resistance development in the target pathogens, in practice fungicides may be applied also when the disease is already present, a situation that commonly occurs in the field. The aim of this work was to study the curative activity of several fungicides toward P. infestans to determine their efficacy when applied after the infection process. Nine trials were performed in greenhouse using potted tomato plants that were treated 24 h after inoculation. Disease severity was assessed three times from the development of the symptoms on the untreated plants and data analysed using a linear mixed model. Differences in post-infection control between the different chemical classes were found. Metalaxyl-M and cymoxanil showed the best curative activity while among the CAA fungicides, a good efficacy was expressed by dimethomorph. Interestingly, evidence of synergy between active ingredients having different modes of action was observed such as in the mixtures containing dimethomorph?+?ametoctradin, dimethomorph?+?pyraclostrobin and fosetyl-Al?+?propamocarb. This study provided useful information on the post-infection activity of some fungicides used to control tomato late blight and should be taken into account to perform more in depth studies at the field level and to improve the management strategies of the disease.     

Surface energy flux measurements in a flooded and an aerobic rice field using a single eddy-covariance system

About 90 % of the global rice production takes place in Asia, while European production is quantitatively modest. Italy is the Europe’s leading producer, with over half of total production concentrated in a large, traditional paddy rice area in the north of the country. High irrigation requirement for continuous flooding encourages the adoption of water saving techniques. In 2013, an intense monitoring activity was conducted on two fields characterized by continuous flooding and intermittent irrigation regimes, with the aim of comparing their agronomical and hydrological effects, including their influence on the energy balance. An eddy covariance station was installed on the levee between the two fields, to monitor latent (LE) and sensible (H) heat fluxes as a function of wind direction. Additionally, the fields were instrumented with net radiometers, soil heat flux (G) plates, thermistors, tensiometers, and multilevel moisture probes. Three footprint models were applied to determine position and size of the footprint area at each monitoring time step, providing similar results. Two half-hourly turbulent fluxes datasets were obtained, one for each irrigation regime, each one comprising about 10 % of the daytime time steps over the agricultural season. The reliability of the monitoring performed with a single EC station was confirmed by the energy balance closure (H + LE versus R_n-G), showing an imbalance lower than 10 % for both the regimes. A detailed analysis of the effect of the storage terms on the ground heat flux estimation and a more thorough analysis of the radiation balance for the two plots were also performed.     

Development of a multiplex RT-PCR assay for simultaneous detection of the major viruses that affect rainbow trout (Oncorhynchus mykiss)

The major viral diseases that affect rainbow trout (Oncorhynchus mykiss) are viral haemorrhagic septicaemia, infectious haematopoietic necrosis, infectious pancreatic necrosis and sleeping disease. In the presented study, we developed a multiplex RT-PCR (mRT-PCR) assay for the simultaneous detection of these four rainbow trout viruses in a single assay. The choice of primers was carried out based on the expected size of the fragments, the temperature and time required for the amplification, and the specificity for the target sequence. Firstly, the method was optimised using reference strains of viral haemorrhagic septicaemia virus (VHSV), infectious haematopoietic necrosis virus (IHNV), infectious pancreatic necrosis virus (IPNV) and sleeping disease virus (SDV) cultivated with permissive cell culture lines; subsequently, the method was used for the identification of these viral infections in rainbow trout samples. Twenty-two samples of rainbow trout, clinically suspected of having viruses, were analysed by the developed method to detect the presence of the four viruses, by directly analysing the animal tissues. The mRT-PCR method was able to efficiently detect the viral RNA in infected cell culture supernatants and in tissue samples, highlighting the presence of single infections as well as co-infections in rainbow trout samples. VHSV/SDV and IHNV/SDV co-infections were demonstrated for the first time in rainbow trout. The mRT-PCR method was revealed to be an accurate and fast method to support traditional diagnostic techniques in the diagnosis of major viral diseases of rainbow trout.     

Coupling an oceanographic model to a Fishery Observing System through mixed models: the importance of fronts for anchovy in the Adriatic Sea

Anchovy, Engraulis encrasicolus, forms the basis of Italian small pelagic fisheries in the Adriatic Sea. The strong dependence of this stock on environmental factors and the consequent high variability makes the dynamics of this species particularly complicated to model. Weekly geo‐referenced catch data of anchovy obtained by means of a Fishery Observing System (FOS) from 2005 to 2011 were referred to a 0.2 × 0.2 degree grid (about 20 km²) and associated with the environmental parameters calculated by a Regional Ocean Modelling System, AdriaROMS. Generalized Additive Mixed Models (GAMM) with and without random effects were used to identify a relationship between abundance in the catch and oceanographic conditions. The outcomes of models with no random effects, with random vessel effects and with the random vessel and random week‐of‐the‐year effects were examined. The GAMM incorporating a random vessel and week‐of‐the‐year effect were selected as the best model on the basis of the Akaike information criteria (AIC). This model indicated that catches (abundance) of anchovy in the Adriatic Sea correlate well with low temperatures, salinity fronts and sea surface height, and allowed the identification of areas where high concentrations of this species are most likely to occur. The results of this study demonstrate that GAMM are a useful tool to combine geo‐referenced catch data with oceanographic variables and that the use of a mixed‐model approach with spatial and temporal random effects is an effective way to depict the dynamics of marine species.     

Detection and identification of beta-lactam residues in milk using a hybrid biosensor

A novel application of a hybrid biosensor is here employed as an analytical method for the detection and presumptive identification of beta-lactam residues in milk. The method is based on measurements of carbon dioxide (CO2), the production of which is related to the microbial growth of the test microorganism Bacillus stearothermophilus var. calidolactis. The presence of beta-lactams in milk inhibits microbial growth and, consequently, the CO2 production rate. The analysis is based on the variation of CO2 between a milk sample spiked with beta-lactams and a twin milk sample containing beta-lactams plus a broad spectrum beta-lactamase, using an electrochemical device of biosensor. A blank milk sample is included as control. The result is obtained starting from the first 120 min. Moreover, the ability to recognize all of the beta-lactams speeds the total time of analysis when chemical identification and quantification are required. The analytical method appears to be adequate for milk control for qualitative screening purposes, complying with the requirements stated in Decision 2002/657/EC.     

The origin of rhizomania resistance in sugar beet

In the last 35 years, breeding has greatly reduced the damages caused by rhizomania in sugar beet crops. After the first encouraging results using the Alba genotypes, the cultivar Rizor represented a substantial step forward and has given good yield improvement in diseased fields in many parts of the world. The original variety and subsequent improved versions continued to offer good performances for about a decade, after which it was surpassed by other hybrids derived in part from the Rizor itself. Further progress in terms of sugar production became possible in 1986, when the Holly monogerm lines were released in USA and Europe. In spite of the incomplete information about the genealogy of the first resistant materials,many evidences and the molecular analyses on the different genotypes suggest a possible common progenitor and lineage. The resistant cultivars have kept the yield at an adequate level, allowing cultivation to continue in countries where the disease has reached epidemic proportions. The case of rhizomania resistance in sugar beet can therefore be considered as one of the most important achievements in plant breeding. This revised version was published online in August 2006 with corrections to the Cover Date.     

Sedative effects of intramuscular alfaxalone in pet guinea pigs (Cavia porcellus)

Objective

To evaluate the efficacy and side effects of alfaxalone administered intramuscularly (IM) as a sedative agent in guinea pigs undergoing survey radiographs.