Long-Term Brain Organotypic Cultures of the Turquoise Killifish Nothobranchius furzeri.

Organotypic culture is a well-established method for culturing ex vivo tissue samples. The advantages of culturing tissue slices for prolonged time periods ex vivo are numerous and consist primarily of the maintenance of the overall in vivo architecture of the isolated sample, the lack of the ematoencephalic barrier, and the ease of pharmacological treatments and interventions that can be conducted under controlled conditions as in in vitro systems such as cell cultures. Given the extremely short life span of Nothobranchius furzeri and the emergence of aging signs only after a few months of life, it is of particular interest to establish this protocol for N. furzeri as a potential method to study brain aging ex vivo.

Quantification of noradrenergic-, dopaminergic-, and tectal-neurons during aging in the short-lived killifish Nothobranchius furzeri.

Parkinson's disease (PD) is characterized by phosphorylation and aggregation of the protein α-Synuclein and ensuing neuronal death progressing from the noradrenergic locus coeruleus to midbrain dopaminergic neurons. In 2019, Matsui and colleagues reported a spontaneous age-dependent degeneration of dopaminergic neurons and an even greater neurodegeneration of the noradrenergic neurons in the short-lived killifish Nothobranchius furzeri. Given the great possible relevance of a spontaneous model for PD, we assessed neurodegeneration of noradrenergic and dopaminergic neurons in two further laboratory strains of N. furzeri. We implemented, for the first time in N. furzeri, a whole-brain clarification technique and proceeded to entire 3D nuclei reconstruction to quantify total cell numbers in two different stains of N. furzeri. In both strains, we observed that age-dependent neurodegeneration is limited to the locus coeruleus and does not involve the posterior tuberculum. We also applied 3D counting to the optic tectum, an area of active adult neurogenesis, and detected an increase of neurons with age. Our results confirm age-dependent neurodegeneration of noradrenergic neurons, a condition reminiscent of the presymptomatic stage of PD indicating that N. furzeri could be used in the future to identify modifying factors for age-dependent neurodegeneration and open the intriguing possibility that natural genetic variation may influence the susceptibility of dopaminergic neurons.

Virulence Genes of Pathogenic Escherichia coli in Wild Red Foxes (Vulpes vulpes).

Different pathotypes of Escherichia coli can cause severe diseases in animals and humans. Wildlife may contribute to the circulation of pathogenic pathotypes, including enteropathogenic E. coli (EPEC), Shiga toxin-producing E. coli (STEC), and enterohemorrhagic E. coli (EHEC). This study analyzed 109 DNA samples previously extracted from fecal specimens collected from red foxes (Vulpes vulpes) to detect E. coli virulence genes eaeA, hlyA, stx1, and stx2, that characterize the EPEC, STEC, and EHEC strains. Thirty-one (28.4%) samples were positive for at least one investigated virulence gene: eaeA gene was detected in 21 (19.2%) samples, hlyA in 10 (9.1%), stx1 in 6 (5.5%), and stx2 in 4 (3.6%). Nine DNA samples resulted positive for two or three virulence genes: five (4.6%) samples were positive for eaeA and hlyA genes, two (1.8%) for eaeA and stx1, one (0.9%) for hlyA and stx1, one (0.9%) for eaeA, hlyA and stx2. Red foxes seem to be involved in the epidemiology of these infections and their role could be relevant because they may be source of pathogenic E. coli for other wild animals, as well as domestic animals and humans.

ß-Glucans as Dietary Supplement to Improve Locomotion and Mitochondrial Respiration in a Model of Duchenne Muscular Dystrophy.

Duchenne muscular dystrophy (DMD) is a severe X-linked neuromuscular childhood disorder that causes progressive muscle weakness and degeneration. A lack of dystrophin in DMD leads to inflammatory response, autophagic dysregulation, and oxidative stress in skeletal muscle fibers that play a key role in the progression of the pathology. ß-glucans can modulate immune function by modifying the phagocytic activity of immunocompetent cells, notably macrophages. Mitochondrial function is also involved in an important mechanism of the innate and adaptive immune responses, owing to high need for energy of immune cells. In the present study, the effects of 1,3-1,6 ß-glucans on five-day-old non-dystrophic and dystrophic (sapje) zebrafish larvae were investigated. The effects of the sonication of ß-glucans and the dechorionation of embryos were also evaluated. The results showed that the incidence of dystrophic phenotypes was reduced when dystrophic embryos were exposed to 2 and 4 mg L-1 of 1,3-1,6 ß-glucans. Moreover, when the dystrophic larvae underwent 8 mg L-1 treatment, an improvement of the locomotor performances and mitochondrial respiration were observed. In conclusion, the observed results demonstrated that 1,3-1,6 ß-glucans improve locomotor performances and mitochondrial function in dystrophic zebrafish. Therefore, for ameliorating their life quality, 1,3-1,6 ß-glucans look like a promising diet supplement for DMD patients, even though further investigations are required.

Effect of 1,3-1,6 ß-Glucan on Natural and Experimental Deformed Wing Virus Infection in Newly Emerged Honeybees (Apis mellifera ligustica).

The Western Honeybee is a key pollinator for natural as well as agricultural ecosystems. In the last decade massive honeybee colony losses have been observed worldwide, the result of a complex syndrome triggered by multiple stress factors, with the RNA virus Deformed Wing Virus (DWV) and the mite Varroa destructor playing crucial roles. The mite supports replication of DWV to high titers, which exert an immunosuppressive action and correlate with the onset of the disease. The aim of this study was to investigate the effect of 1,3-1,6 ß-glucan, a natural innate immune system modulator, on honeybee response to low-titer natural and high-titer experimental DWV infection. As the effects exerted by ß-glucans can be remarkably different, depending on the target organism and the dose administered, two parallel experiments were performed, where 1,3-1,6 ß-glucan at a concentration of 0.5% and 2% respectively, was added to the diet of three cohorts of newly emerged honeybees, which were sampled from a Varroa-free apiary and harboured a low endogenous DWV viral titer. Each cohort was subjected to one of the following experimental treatments: no injection, injection of a high-copy number DWV suspension into the haemocel (experimental DWV infection) or injection of PBS into the haemocoel (physical injury). Control bees fed a ß-glucan-free diet were subjected to the same treatments. Viral load, survival rate, haemocyte populations and phenoloxidase activity of each experimental group were measured and compared. The results indicated that oral administration of 0.5% ß-glucan to naturally infected honeybees was associated with a significantly decrease of the number of infected bees and viral load they carried, and with a significant increase of the survival rate, suggesting that this natural immune modulator molecule might contribute to increase honeybee resistance to viral infection.

Photopolymerized Network Polysiloxane Films with Dangling Hydrophilic/Hydrophobic Chains for the Biofouling Release of Invasive Marine Serpulid Ficopomatus enigmaticus.

Novel photopolymerized network films based on a polysiloxane matrix containing varied amounts of polyoxyethylene (P3) or perfluorohexylethyl (F) dangling side chains were investigated. For films containing less than 10 wt % P3 and F, the wettability and elastic modulus were similar to those of the photopolymerized network matrix. However, angle-resolved X-ray photoelectron spectroscopy measurements proved that the surface of films with F dangling chains was highly enriched in fluorine depending on both the amount of P3 and F and their relative ratio in the films. The biological performance of the films was evaluated against a new widespread and invasive marine biofoulant, the serpulid Ficopomatus enigmaticus. The diatom Navicula salinicola was also assayed as a conventional model organism for comparison. Films richer in P3 better resisted the settlement and promoted the release of calcified tubeworms of F. enigmaticus.

Parasitic infections in an organic grazing cattle herd in Tuscany using geographic information systems to determine risk factors.

An organic grazing cattle herd in Tuscany (Italy) was monitored for parasites between 2002 and 2006. Every two to three months, faecal samples from cattle of different breeds and age were collected and examined for endoparasites, using both qualitative and quantitative parasitological techniques. Several environmental parameters were monitored and data on biodiversity and field margin biodiversity of grazing areas were also collected. All data were geo-referenced and plotted on a vectorial map using geographic information systems (GIS) software. Soil was classified as silt and clay/sand. The hydraulic drainage was poor and water pooling was observed frequently. The biodiversity of field margins was relatively high. Cattle were infected by coccidia, gastrointestinal nematodes, cestodes and trematodes. Prevalence and intensity of infestations were highly variable. In most cases, this variability was related to cattle breed, age, season and meteorological data. The Pisana breed was most commonly infected by Fasciola hepatica and Paramphistomidae. These infestations were associated with more frequent flooding and water pooling in the areas grazed by this breed.