Niche partitioning between sympatric wild canids: the case of the golden jackal (Canis aureus) and the red fox (Vulpes vulpes) in north-eastern Italy.

BACKGROUND: Two coexisting species with similar ecological requirements avoid or reduce competition by changing the extent of their use of a given resource. Numerous coexistence mechanisms have been proposed, but species interactions can also be aggressive; thus, generally a subordinate species modifies its realized niche to limit the probability of direct encounters with the dominant species. We studied niche partitioning between two sympatric wild canids in north-eastern Italy: the golden jackal and the red fox, which, based on competition theories, have a high potential for competition. We considered four main niche dimensions: space, habitat, time, and diet. RESULTS: We investigated three study areas monitoring target species populations from March 2017 to November 2018 using non-invasive monitoring techniques. Red fox presence was ascertained in every study area, while golden jackal presence was not ascertained in one study area, where we collected data regarding wolf presence. Considering the two target species, we observed partial diet partitioning based on prey size, with the golden jackal mainly feeding on wild ungulates and the red fox mainly feeding on small mammals. The two canids had an extensive temporal overlap along the diel cycle, having both predominant crepuscular and nocturnal activity patterns, but marked spatial partitioning and differential use of habitats. The golden jackal proved to be specialist concerning the habitat dimension, while the red fox resulted completely generalist: the former selected less human-modified habitats and avoided intensively cultivated lands, while the latter was present in all habitats, including intensively cultivated lands. CONCLUSIONS: The observed partitioning might be due partially to some ecological adaptations (e.g. specialist vs. generalist use of resources) and specific behaviours (e.g. cooperative vs. solitary hunting) and partially to the avoidance response of the red fox aimed at reducing the probability of direct encounters with the golden jackal.

Advances in molecular tools for the use of Zygosaccharomyces bailii as host for biotechnological productions and construction of the first auxotrophic mutant.

The nonconventional yeast Zygosaccharomyces bailii has been proposed as a new host for biotechnological processes due to convenient properties such as its resistance to high sugar concentrations, relatively high temperatures and especially to acidic environments. We describe a series of new expression vectors specific for Z. bailii and the resulting improvements in production levels. By exploiting the sequences of the endogenous plasmid pSB2, 2microm-like multicopy vectors were obtained, giving a fivefold increase in production. A specific integrative vector was developed which led to 100% stability in the absence of selective pressure; a multiple-integration vector was constructed, based on an rRNA gene unit portion cloned and sequenced for this purpose, driving the insertion of up to 80 copies of the foreign construct. Moreover, we show the construction of the first stable auxotrophic mutant of Z. bailii, obtained by targeted gene deletion applied to ZbLEU2. The development of molecular tools for the Z. bailii manipulation has now reached a level that may be compatible with its industrial exploitation; the production of organic acids is a prominent field of application.

Cloning of the Zygosaccharomyces bailii GAS1 homologue and effect of cell wall engineering on protein secretory phenotype.

BACKGROUND: Zygosaccharomyces bailii is a diploid budding yeast still poorly characterized, but widely recognised as tolerant to several stresses, most of which related to industrial processes of production. Because of that, it would be very interesting to develop its ability as a cell factory. Gas1p is a beta-1,3-glucanosyltransglycosylase which plays an important role in cell wall construction and in determining its permeability. Cell wall defective mutants of Saccharomyces cerevisiae and Pichia pastoris, deleted in the GAS1 gene, were reported as super-secretive. The aim of this study was the cloning and deletion of the GAS1 homologue of Z. bailii and the evaluation of its deletion on recombinant protein secretion. RESULTS: The GAS1 homologue of Z. bailii was cloned by PCR, and when expressed in a S. cerevisiae GAS1 null mutant was able to restore the parental phenotype. The respective Z. bailii Deltagas1 deleted strain was obtained by targeted deletion of both alleles of the ZbGAS1 gene with deletion cassettes having flanking regions of approximately 400 bp. The morphological and physiological characterization of the Z. bailii null mutant resulted very similar to that of the corresponding S. cerevisiae mutant. As for S. cerevisiae, in the Z. bailii Deltagas1 the total amount of protein released in the medium was significantly higher. Moreover, three different heterologous proteins were expressed and secreted in said mutant. The amount of enzymatic activity found in the medium was almost doubled in the case of the Candida rugosa lipase CRL1 and of the Yarrowia lipolytica protease XPR2, while for human IL-1beta secretion disruption had no relevant effect. CONCLUSIONS: The data presented confirm that the engineering of the cell wall is an effective way to improve protein secretion in yeast. They also confirmed that Z. bailii is an interesting candidate, despite the knowledge of its genome and the tools for its manipulation still need to be improved. However, as already widely reported in literature, our data confirmed that an "always working" solution to the problems related to recombinant protein production can be hardly, if never, found; instead, manipulations have to be finely tuned for each specific product and/or combination of host cell and product.