Canine fertility: The consequences of selection for special traits

Pedigree dogs and cats are bred aiming to conform breed standards with very poor consideration for breeding stock fertility. At the same time, the genetic asset underlining reproductive traits could be effectively analysed like in other species under selection. The definition of selection targets is very important in breeding protocols determination. The aim of the present work is to present an overview of the different correlations between reproduction and genetics, starting from selection procedure and inbreeding coefficient moving to genomic and the application of SNPs and GWAS on population study and identification of genes involved in phenotypical variation of reproductive traits in dogs. Particular relevance has been given to the concept of inbreeding which effects on canine reproduction have been presented. The use of genomic information in inbreeding coefficient calculation can be considered an improved effective procedure in the evaluation of the genetic variability loss in canine population and its negative effects on reproductive traits.     

Post-mortem recovery,in vitro maturation and fertilization of fallow deer (Dama dama,Linnaeus 1758) oocytes collected during reproductive and no reproductive season

Habitat degradation leads to small and fragmented populations, lower genetic variability and fertility overtime. Assisted reproductive techniques represent important tools to cope with the dramatic loss of biodiversity. Fallow deer (Dama dama), beyond its high commercial value and wide distribution, may represent the most suitable model to study endangered cervids. In this study, oocytes were recovered post-mortem from fallow deer during the breeding and no breeding seasons and were in vitro matured (IVM). The ability of cryopreserved thawed sperm samples recovered by electroejaculation from four adult males was tested by in vitro fertilization of IVM oocytes. The number of oocytes collected per ovary did significantly vary across seasons from 6.2 ± 0.92 during breeding season to 10.4 ± 1.26 during no breeding season (p = .006). Oocytes collected during the breeding season showed higher in vitro fertilization rate compared to the no breeding season (p = .045). However, no embryos reached the blastocyst stage. Semen samples obtained by electroejaculation were successfully cryopreserved, although the cryopreservation process negatively affected most kinetic parameters, mainly at 2 hr post-thawing. Moreover, the percentage of rapid spermatozoa significantly decreased between fresh samples and at 2 hr post-thawing, whereas the percentage of slow spermatozoa increased across the same period (p < .05). Our study provides the logistic steps for the application of assisted reproductive techniques in fallow deer and might be of great interest for genetic resource bank planning.     

In Vitro Bioactivity of Astaxanthin and Peptides from Hydrolisates of Shrimp (Parapenaeus longirostris) By-Products: From the Extraction Process to Biological Effect Evaluation,as Pilot Actions for the Strategy “From Waste to Profit”

Non-edible parts of crustaceans could be a rich source of valuable bioactive compounds such as the carotenoid astaxanthin and peptides, which have well-recognized beneficial effects. These compounds are widely used in nutraceuticals and pharmaceuticals, and their market is rapidly growing, suggesting the need to find alternative sources. The aim of this work was to set up a pilot-scale protocol for the reutilization of by-products of processed shrimp, in order to address the utilization of this valuable biomass for nutraceutical and pharmaceuticals application, through the extraction of astaxanthin-enriched oil and antioxidant-rich protein hydrolysates. Astaxanthin (AST) was obtained using “green extraction methods,” such as using fish oil and different fatty acid ethyl esters as solvents and through supercritical fluid extraction (SFE), whereas bioactive peptides were obtained by protease hydrolysis. Both astaxanthin and bioactive peptides exhibited bioactive properties in vitro in cellular model systems, such as antioxidant and angiotensin I converting enzyme (ACE) inhibitory activities (IA). The results show higher astaxanthin yields in ethyl esters fatty acids (TFA) extraction and significant enrichment by short-path distillation (SPD) up to 114.80 ± 1.23 µg/mL. Peptide fractions of <3 kDa and 3–5 kDa exhibited greater antioxidant activity while the fraction 5–10 kDa exhibited a better ACE-IA. Lower-molecular-weight bioactive peptides and astaxanthin extracted using supercritical fluids showed protective effects against oxidative damage in 142BR and in 3T3 cell lines. These results suggest that “green” extraction methods allow us to obtain high-quality bioactive compounds from large volumes of shrimp waste for nutraceutical and pharmaceutical applications.     

Soil humic substances stimulate proton release by intact oat seedling roots

The effect of a soil humic fraction (HS) on proton extrusion into deionized water by intact oat seedling roots was studied. In the presence of HS, at concentration of 10 μg organic carbon (C) mL^‐1, a clear stimulation of acidification of the outer medium by the roots was observed after three to four hours of incubation. The addition of 0.5 mM vanadate to the solution bathing the roots drastically reduced the net proton extrusion, either in the presence or absence of HS, suggesting the involvement of the plasma membrane H⁺‐ATPase in the stimulation of the acidification of the outer medium by oat roots. The release of potassium (K) from the roots into deionized water was also monitored concomitantly to the proton extrusion. The loss of endogenous K from the roots was similar in the presence or absence of HS, while the recovery of the cation was slower in the presence of the humic fraction. However, after reabsorption of the released K, no net acidification was observed in control roots, while HS‐treated roots significantly decreased the pH of the deionized water. The addition of 3 mM K to the external medium greatly enhanced the proton release from roots, while the presence of humic substances reduced the magnitude of the stimulation by K. When K was supplied at a concentration closer to that encountered in the soil (<0.1 mM), HS significantly stimulated proton release. The effect of HS on root extracellular acidification supports the idea of a role of soil humic substances on plant nutrition via interaction with cell membrane functions.     

First report of QoI resistance in Italian population of Pyricularia oryzae

Journal of Plant Diseases and Protection - Rice blast disease, caused by Pyricularia oryzae Cavara, is considered the most destructive rice disease in Italy. Quinone outside inhibitor (QoI)...     

Bacterial community associated with the ambrosia beetle Platypus cylindrus on declining Quercus suber trees in the Alentejo region of Portugal

In Portugal, the oak pinhole borer Platypus cylindrus and its mycobiota have been associated with cork oak (Quercus suber) death, but no knowledge exists regarding the associated bacterial community. However, it is known that some bacteria are important for ambrosia beetle symbiosis and play a role in oak tree health. To explore the bacteria associated with this beetle and its host, with the ultimate goal of highlighting potential roles in oak decline, this study used a culture-dependent approach for strain isolation and phylogenetic identification using 16S rRNA gene sequencing and multilocus sequence analysis (MLSA). The bored galleries of different cork oak trees from a cork stand in Alentejo, together with the body and mycangia of adult beetles, were investigated. The samples revealed a diverse community comprising 500 isolates with 64 distinct types of bacterial colonies. Sixty-eight strains were selected for sequencing and used for phylogenetic analysis, 40 from wood galleries and 28 from beetles. Thirty-two genera of bacteria were identified, 18 of which were described for the first time within oak–beetle interactions. Major taxonomic groups were Actinobacteria in beetles and Enterobacterales in wood galleries. Although specific oak bacterial pathogens were not detected, a group of distinct strains detected in wood galleries, potentially belonging to a new Pectobacteriaceae species, were able to produce mild symptoms on cork oak plantlets. This study reports for the first time the biodiversity of culturable bacteria associated with the Q. suber–P. cylindrus interaction, their relevance to both organisms and the possible contribution to oak decline.     

Microbial dynamics after adding bovine manure effluent together with a nitrification inhibitor (3,4 DMPP) in a microcosm experiment

The application of animal manure effluents in agriculture in combination with nitrification inhibitors should be beneficial for nutrient recycling, soil quality, plant productivity, and greenhouse gas emission and offer economic advantages to make them an alternative to conventional fertilizers. The present study aims to estimate the effects of the addition of bovine manure effluent alone or together with a nitrification inhibitor (3,4-dymethylpyrazol-phosphate (3,4 DMPP)) on the microbial community dynamics in a Mediterranean soil in an incubation experiment over 28 days. The application of the bovine manure effluent increased respiration, microbial biomass carbon, fungal and bacterial growth, and enzyme activities and changed the microbial community structure evaluated by the phospholipid fatty acid pattern. Adding the bovine manure effluent together with the nitrification inhibitor, although partly negating the positive effect of the effluent on soil microbial activity, still resulted in higher or similar growth and activity as in the control. Our results indicate that the addition of the nitrification inhibitor 3,4 DMPP together with a bovine manure effluent could be a promising solution to control the animal manure effluent application effects on soil microbiological properties and microbial dynamics, as well as counteracting direct inhibiting effects of 3,4 DMPP on the soil heterotrophic community.     

Map- vs. homology-based cloning for the recessive gene ol-2 conferring resistance to tomato powdery mildew

The recessive gene ol-2 confers papilla-associated and race-non-specific resistance to tomato powdery mildew caused by Oidium neolycopersici. In order to facilitate marker assisted selection (MAS) in practical breeding programmes, we identified two simple sequence repeat (SSR) markers and one cleaved amplified polymorphic sequence (CAPS) marker which are linked to the resistance locus and co-dominantly inherited. Aiming to provide a base for ol-2 positional cloning, we used a large segregating F₂ population to merge these markers with all the ol-2 linked amplified fragment length polymorphism (AFLP^®) markers previously identified in an integrated genetic map. By screening a tomato bacterial artificial chromosome (BAC) library, we detected two BAC clones containing two expressed sequence tags (ESTs) homologous to the gene mlo, responsible for powdery mildew resistance in barley, as well as an ol-2-linked marker. Chromosomal mapping by Fluorescence in situ Hybridization (FISH) revealed major signals of the two BAC DNAs in the pericentromeric heterochromatin of the short arm of chromosome 4, in the same region where the ol-2 gene was previously mapped. The genetic and cytogenetic co-localisation between ol-2 and tomato mlo-homologue(s), in addition to the similarity of ol-2 and mlo resistances for both genetic and phytopathological characteristics, suggests that ol-2 is likely a mlo-homologue. Thus, a homology-based cloning approach could be more suitable than positional cloning for ol-2 isolation.     

Biochemical Characterization of Cassiopea andromeda (Forsskål, 1775), Another Red Sea Jellyfish in the Western Mediterranean Sea

Increasing frequency of native jellyfish proliferations and massive appearance of non-indigenous jellyfish species recently concur to impact Mediterranean coastal ecosystems and human activities at sea. Nonetheless, jellyfish biomass may represent an exploitable novel resource to coastal communities, with reference to its potential use in the pharmaceutical, nutritional, and nutraceutical Blue Growth sectors. The zooxanthellate jellyfish Cassiopea andromeda, Forsskål, 1775 (Cnidaria, Rhizostomeae) entered the Levant Sea through the Suez Canal and spread towards the Western Mediterranean to reach Malta, Tunisia, and recently also the Italian coasts. Here we report on the biochemical characterization and antioxidant activity of C. andromeda specimens with a discussion on their relative biological activities. The biochemical characterization of the aqueous (PBS) and hydroalcoholic (80% ethanol) soluble components of C. andromeda were performed for whole jellyfish, as well as separately for umbrella and oral arms. The insoluble components were hydrolyzed by sequential enzymatic digestion with pepsin and collagenase. The composition and antioxidant activity of the insoluble and enzymatically digestible fractions were not affected by the pre-extraction types, resulting into collagen- and non-collagen-derived peptides with antioxidant activity. Both soluble compounds and hydrolyzed fractions were characterized for the content of proteins, phenolic compounds, and lipids. The presence of compounds coming from the endosymbiont zooxanthellae was also detected. The notable yield and the considerable antioxidant activity detected make this species worthy of further study for its potential biotechnological sustainable exploitation.