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.     

Molecular adaptation of barley to cold and drought conditions

Summary Molecular adaptation to cold and drought involves a series of biochemical and molecular changes leading plants to improve their winter hardiness or drought resistance.We are interested to study the molecular basis of cold acclimation and drought response of barley to survive under stress. Several genes regulated by low temperatures and sometimes by drought have been isolated from the barley genome. In this review the most significant results of our recent work will be presented and discussed.The protein encoded by cDNA clone pt59 and induced in barley by cold was over-expressed in E. coli to produce the matching antibody, which in vivo recognizes a cold-induced protein of 14 kDa (COR14). The COR14 is stored in amounts only slightly greater in the cold resistant Onice than in the susceptible Gitane, although the former has a higher induction-temperature threshold of COR14 than the latter. This fact is an evolutionary advantage enabling the resistant varieties in the field to prepare for the cold well ahead of the susceptible ones.Two other cDNA clones, paf93 and cdr29, are regulated by low temperature and drought stress but not by exogenous ABA treatment. Indeed during the early stage of dehydration, the mRNAs are expressed before the induction of known ABA regulated genes such as dehydrins and when only a small increase occurs in ABA content. The sequence analysis revealed that paf93 encodes for a protein homologous to the cold-regulated protein COR47 of Arabidopsis, whereas cdr29 represents a plant gene homologous to yeast and mammalian sequences coding for acyl-Coenzyme A oxidase.Abbreviations ABA abscisic acid - COR cold regulated     

Characterization of Italian lentil (Lens culinaris Medik.) germplasm by agronomic traits, biochemical and molecular markers

Genetic relationships, agronomic, nutritional and technological traits of ten Italian landraces, two improved lines and two cultivars of lentil (Lens culinaris Medik.) were investigated using a multi-disciplinary approach. Seed storage proteins, used as biochemical markers, were able to detect polymorphisms with variability mainly related to the polypeptide abundance. Microsatellite (SSR) molecular markers provided very useful information on genetic variation and relationships among landraces, with polymorphic fragments able to discriminate all the accessions. Lentil landraces were grouped in different clusters and sub-clusters principally on the basis of their geographical origin. The highest levels of genetic diversity were observed for lentils from ‘Castelluccio di Norcia’, ‘Colliano’ and ‘Villalba’. Field trials, performed in two locations of Southern Italy, revealed a high influence of location on yield. Comparing performances at both tested locations, the best landraces were ‘Linosa’ and ‘Valle di Nevola’ suggesting that these have the highest adaptability. Technological and nutritional data together with the agronomic ones evidenced that ‘Linosa’ lentil is the best landrace, however also ‘San Gerardo’ deserves some attention.     

Agronomic performance and combining ability estimates of yellow maize inbred lines under adequate and deficit moisture conditions

Journal of Crop Science and Biotechnology - Breeding for drought tolerance and increased grain yield is vital in mitigating the threat posed by recurrent drought stress on maize production, as well...     

Sensitive diagnosis of bovine tuberculosis in a farmed cervid herd with use of an MPB70 protein fluorescence polarization assay

After histopathological examination of a lesion found in a herd member returned a diagnosis of mycobacteriosis, a farmed herd (n = 47) of elk (Cervus elaphus nelsoni) and red deer (C. elaphus elaphus) was investigated for bovine tuberculosis with a battery of antemortem and postmortem diagnostic tests. Every animal was tested with the mid-cervical tuberculin skin test; all 47 had negative results. All of the 16 adult animals and 15 of the 31 calves (approximately 2-years-old) were blood-tested with a lymphocyte stimulation test (LST) and a fluorescence polarization assay (FPA), which detects antibody to the MPB70 protein antigen. At necropsy of the 31 blood-tested animals, tissues were harvested for histopathological examination and culture of mycobacteria. Mycobacterium bovis was isolated from 16 of the 31 animals, and a scotochromogen was also isolated from 1 of the 16 whose tissues yielded M. bovis. Each of these 16 animals, 15 of which were calves, also received a histopathological diagnosis of mycobacteriosis. Other species of mycobacteria, including those belonging to the M. avium and M. terrae complexes, were isolated from an additional 7 animals. The FPA was scored “positive” or “suspect” for 16 animals, 13 (81%) of which were culture-positive for M. bovis. The other 3 animals that were culture-positive for M. bovis had negative FPA results. Of the 3 FPA-positive or FPA-suspect animals that were culture-negative, 2 were suspected to have mycobacteriosis on the basis of the histopathological examination. The 7 animals from which Mycobacterium species other than M. bovis were cultured were all FPA-negative. The only animal with positive LST results was also FPA-positive and culture-positive for M. bovis. The M. bovis isolates had an identical spoligotype pattern, with an octal code of 664073777777600. This is the first report of the isolation and identification of this strain type in Canada.