The Application of 3D Anatomy for Teaching Veterinary Clinical Neurology.

Neuroanatomy is always a challenging topic for veterinary students. It is widely accepted that understanding the anatomy of the central nervous system (CNS) is essential to explain many of the pathological processes that affect the brain. Although its study has varied over time to achieve this goal, in human and veterinary medicine it is difficult to find a teaching method that associates normal anatomy with pathological alterations of the brain. For the first time, we have created an educational tool that combines neuroanatomy and neuropathology, using different magnetic resonance (MR) images as a basis and EspINA software as analyzer, to obtain segmented structures and 3D reconstructions of the dog brain. We demonstrate that this combination is an optimal tool to help anatomists to understand the encephalon, and additionally to help clinicians to recognize illness including a multitude of neurological problems. In addition, we have tried to see whether photogrammetry, which is a common technique in other sciences, for example geology, could be useful to teach veterinary neuroanatomy. Although we still need further investigations, we have been able to generate 3D reconstructions of the whole brain, with very promising results to date.

Ligilactobacillus salivarius MP100 as an Alternative to Metaphylactic Antimicrobials in Swine: The Impact on Production Parameters and Meat Composition.

The metaphylactic use of antimicrobials in swine farms contributes to the emergence of antibiotic-resistant bacteria, which constitutes a major challenge for public health. Alternative strategies are required to eradicate their routine use. In a previous study, metaphylactic antimicrobials were replaced by the administration of Ligilactobacillus salivarius MP100 to sows and piglets for two years. This practice positively modified the fecal microbiota and metabolic profiles in the farm. In this work, the farm dataset was used to compare the productivity-related parameters between a 2-year period of routine metaphylactic antibiotherapy and the first 2 years of a replacement with the probiotic strain. The probiotic period improved these productivity-related parameters, from litter size to growth performance. In addition, samples of Longissimus lumborum, including skin and subcutaneous fat, were obtained from the animals ingesting the probiotic strain and controls (metaphylactic antibiotherapy) and analyzed for their pH, water holding capacity, composition, and metabolic profiling. The probiotic intake did not negatively affect the meat composition and was associated with an increase in inosine concentration and a slight tendency for increasing the intramuscular fat content. These factors are considered as biomarkers of meat quality. In conclusion, the substitution of metaphylactic antimicrobials with the administration of the probiotic strain was associated with beneficial productivity and meat quality outcomes.

Intraventricular hemorrhage induces inflammatory brain damage with blood-brain barrier dysfunction in immature rats.

BACKGROUND: We aimed to characterize a preclinical model of intraventricular hemorrhage-induced brain damage (IVH-BD) in extremely low birth weight newborns (ELBWN), to identify potential therapeutic targets based on its pathophysiology. METHODS: IVH was induced in 1-day-old (P1) Wistar rats by left periventricular injection of clostridium collagenase (PVCC). At P6, P14, and P45 IVH-BD (area of damage, motor and cognitive deficits, Lactate/N-acetylaspartate ratio), white matter injury (WMI: ipsilateral hemisphere and corpus callosum atrophy, oligodendroglial population and myelin basic protein signal reduction), blood-brain barrier (BBB) dysfunction (occludin and Mfsd2a expression, Gadolinium leakage) and inflammation (TNFα, TLR4, NFkB, and MMP9 expression; immune cell infiltration), excitotoxicity (Glutamate/N-acetylaspartate), and oxidative stress (protein nitrosylation) were assessed. Sham animals were similarly studied. RESULTS: IVH-BD leads to long-term WMI, resulting in motor and cognitive impairment, thus reproducing IVH-BD features in ELBWN. BBB dysfunction with increased permeability was observed at P6 and P14, coincident with an increased inflammatory response with TLR4 overexpression, increased TNFα production, and increased immune cell infiltration, as well as increased excitotoxicity and oxidative stress. CONCLUSIONS: This model reproduced some key hallmarks of IVH-BD in ELBWN. Inflammation associated with BBB dysfunction appears as relevant therapeutic target to prevent IVH-BD-induced WMI. IMPACT: Paraventricular injection of clostridium collagenase (PVCC) to 1-day-old Wistar rats uniquely reproduced the neuroimaging, histologic and functional characteristics of intraventricular hemorrhage-induced brain damage (IVH-BD) in extremely low birth weight newborns (ELBWN). PVCC-induced IVH triggered a prolonged inflammatory response associated with blood-brain barrier increased permeability, which in turn facilitates the infiltration of inflammatory cells. Thus, PVCC led to white matter injury (WMI) resulting in long-term motor and cognitive impairment. This model offers a valuable tool to obtain further insight into the mechanisms of IVH-BD in ELBWN and proposes some key therapeutic targets.

Hypothyroidism confers tolerance to cerebral malaria.

The modulation of the host's metabolism to protect tissue from damage induces tolerance to infections increasing survival. Here, we examined the role of the thyroid hormones, key metabolic regulators, in the outcome of malaria. Hypothyroidism confers protection to experimental cerebral malaria by a disease tolerance mechanism. Hypothyroid mice display increased survival after infection with Plasmodium berghei ANKA, diminishing intracranial pressure and brain damage, without altering pathogen burden, blood-brain barrier disruption, or immune cell infiltration. This protection is reversed by treatment with a Sirtuin 1 inhibitor, while treatment of euthyroid mice with a Sirtuin 1 activator induces tolerance and reduces intracranial pressure and lethality. This indicates that thyroid hormones and Sirtuin 1 are previously unknown targets for cerebral malaria treatment, a major killer of children in endemic malaria areas.

Heparin length in the coating of extremely small iron oxide nanoparticles regulates in vivo theranostic applications.

The positive contrast of extremely small iron oxide nanoparticles (ESIONP) in magnetic resonance imaging (MRI) rejuvenates this class of metal nanoparticles (NP).Yet, the current synthesis often lacks the possibility of adjusting the core size (while it is a key element for ESIONP-based MRI contrast behaviour), and also involved multiple complex steps before obtaining a ready-to-use probe for medical applications. In this study, we faced these challenges by applying heparin oligosaccharides (HO) of different lengths as coatings for the preparation of HEP-ESIONP with a one-pot microwave method. We demonstrated that the HO length could control the core size during the synthesis to achieve optimal positive MRI contrast, and that HEP-ESIONP were endowed directly with anticoagulant properties and/or a specific antitumor activity, according to the HO used. Relevantly, positron emission tomography (PET)-based in vivo biodistribution study conducted with 68Ga core-doped HEP-ESIONP analogues revealed significant changes in the probe behaviours, the shortening of HO promoting a shift from hepatic to renal clearance. The different conformations of HO coatings and a thorough in vitro characterisation of the probes' protein coronas provided insight into this crucial impact of HO length on opsonization-mediated immune response and elimination. Overall, we were able to identify a precise HO length to get an ESIONP probe showing prolonged vascular lifetime and moderate accumulation in a tumor xenograft, balanced with a low uptake by non-specific organs and favourable urinary clearance. This probe met all prerequisites for advanced theranostic medical applications with a dual MRI/PET hot spot capability and potential antitumor activity.

Sex Determination in Two Species of Anuran Amphibians by Magnetic Resonance Imaging and Ultrasound Techniques.

The objective of the present study was to evaluate whether gender determination in two amphibian species (Kaloula pulchra and Xenopus laevis) can be reliably carried out by means of magnetic resonance imaging (benchtop magnetic resonance imaging; BT-MRI) or ultrasound (high-resolution ultrasound; HR-US) techniques. Two species of healthy, sexually mature anurans have been used in the present study. Eight Kaloula (blind study) and six Xenopus were used as controls. Magnetic resonance imaging experiments were carried out on a low-field (1 Tesla) benchtop-MRI (BT-MRI) system. HR-US examination was performed with high-resolution equipment. Low-field BT-MRI images provided a clear and quantifiable identification of all the sexual organs present in both genders and species. The HR-US also allowed the identification of testes and ovaries in both species. Results indicate that BT-MRI allowed a very precise sex identification in both anuran species, although its use is limited by the cost of the equipment and the need for anesthesia. HR-US allowed an accurate identification of ovaries of both species whereas a precise identification of testes is limited by the ultrasonographer experience. The main advantages of this technique are the possibility of performing it without anesthesia and the higher availability of equipment in veterinary and zoo institutions.

Vitamin D3 Supplementation in Drinking Water Prior to Slaughter Improves Oxidative Status, Physiological Stress, and Quality of Pork.

The aims of this study were to investigate the effect of vitamin D3 administration in drinking water during lairage time prior to slaughter on physiological stress, oxidative status, and pork quality characteristics. Two experiments were carried out. The first one was performed to examine the effect of vitamin D3 supplementation in drinking water, and the second one to check the effect of supplementation dose (500,000 IU/L vs. 700,000 IU/L). Serum calcium concentration was greater in pigs receiving vitamin D3 in water when compared to the control group. In experiment 1, a 40% α-tocopherol increase in meat from the group supplemented with vitamin D3 (500,000 IU/animal) was observed, that resulted in a tendency of decreased meat malondialdehyde (MDA) values at days 5 and 8 after refrigerated storage. In experiment 2, since water intake was higher (800,000 IU and 1,120,000 IU/animal of vitamin D3 consumption) effects on oxidative status were more profound and vitamin D3 supplementation increased serum α-tocopherol and decreased cortisol and serum TBARS. These effects were also observed in meat; TBARS levels were decreased after 3 days of refrigerated storage. In both experiments meat from pigs that received vitamin D3 in drinking water had a lower proportion of total free-polyunsaturated fatty acids (mainly n-6) when compared to the unsupplemented pigs, and these were positively correlated with TBARS production at day 5 of refrigerated storage (r = 0.53 and 0.38 for experiments 1 and 2, respectively). Meat from pigs receiving vitamin D3 in water showed reduced cohesiveness, gumminess, and chewiness values compared to the control group. The magnetic resonance imaging study of muscle confirmed the effects on water retention with lesser transverse relaxation time in pigs supplemented with vitamin D3. No vitamin D3 dose effect was observed, apart from muscle α-tocopherol concentration that was higher in pigs supplemented with 700,000 compared to those supplemented with 500,000 UI/L.

1H HR-MAS NMR-based metabolomics analysis for dry-fermented sausage characterization.

Proton high-resolution magic angle spinning (1H HR-MAS) nuclear magnetic resonance (NMR) spectroscopy in combination with principal component analysis (PCA) was employed to characterize dry-fermented sausages salchichón type throughout the manufacturing process. 1H HR-MAS NMR metabolite profiling was achieved from a small sample of intact sausage after 0, 2, 4, 7, 11 and 14days of drying. Intriguingly, the obtained results enabled the identification of the three main stages in the traditional production of salchichón. Formulation, fermentation and drying-ripening periods showed distinct and characteristic metabolomic profiles. Compositional changes related to microbial activity, as well as proteolytic and lipolytic phenomena, decisive steps in such a ripening process, could be monitored through the NMR spectra. This study shows the potential of 1H HR-MAS as a rapid method for probing metabolomic profiles and compositional changes during sausages processing.

Automatic ¹H-NMR Screening of Fatty Acid Composition in Edible Oils.

In this work, we introduce an NMR-based screening method for the fatty acid composition analysis of edible oils. We describe the evaluation and optimization needed for the automated analysis of vegetable oils by low-field NMR to obtain the fatty acid composition (FAC). To achieve this, two scripts, which automatically analyze and interpret the spectral data, were developed. The objective of this work was to drive forward the automated analysis of the FAC by NMR. Due to the fact that this protocol can be carried out at low field and that the complete process from sample preparation to printing the report only takes about 3 min, this approach is promising to become a fundamental technique for high-throughput screening. To demonstrate the applicability of this method, the fatty acid composition of extra virgin olive oils from various Spanish olive varieties (arbequina, cornicabra, hojiblanca, manzanilla, and picual) was determined by ¹H-NMR spectroscopy according to this protocol.

Metabolomics of meat exudate: Its potential to evaluate beef meat conservation and aging.

In this study we analyzed the exudate of beef to evaluate its potential as non invasive sampling for nuclear magnetic resonance (NMR) based metabolomic analysis of meat samples. Exudate, as the natural juice from raw meat, is an easy to obtain matrix that it is usually collected in small amounts in commercial meat packages. Although meat exudate could provide complete and homogeneous metabolic information about the whole meat piece, this sample has been poorly studied. Exudates from 48 beef samples of different breeds, cattle and storage times have been studied by (1)H NMR spectroscopy. The liquid exudate spectra were compared with those obtained by High Resolution Magic Angle Spinning (HRMAS) of the original meat pieces. The close correlation found between both spectra (>95% of coincident peaks in both registers; Spearman correlation coefficient = 0.945) lead us to propose the exudate as an excellent alternative analytical matrix with a view to apply meat metabolomics. 60 metabolites could be identified through the analysis of mono and bidimensional exudate spectra, 23 of them for the first time in NMR meat studies. The application of chemometric tools to analyze exudate dataset has revealed significant metabolite variations associated with meat aging. Hence, NMR based metabolomics have made it possible both to classify meat samples according to their storage time through Principal Component Analysis (PCA), and to predict that storage time through Partial Least Squares (PLS) regression.

1H-HRMAS NMR study of cold smoked Atlantic salmon (Salmo salar) treated with E-beam.

(1)H High resolution magic angle spinning (HRMAS) NMR spectroscopic data in combination with principal components analysis and analysis of variance were used to differentiate between irradiated and non-irradiated cold-smoked Atlantic salmon (Salmo salar). NMR profiling was obtained, with a spectral acquisition time of less than 8 min, from a small sample size of intact white salmon muscle, by non-destructive analysis that includes a very simple and rapid sample preparation step. Results obtained enable the use of creatine, trimethylamine oxide and the sum of phosphorylcholine and glycerophosphorylcholine as diagnostic compounds to detect irradiation treatment. This study shows the potential of (1)H-HRMAS to be a rapid method for investigating compositional changes due to food processing as well as to confirm the presence or absence of some bioactive compounds in irradiated samples.

1H-HRMAS NMR study of smoked Atlantic salmon (Salmo salar).

High-resolution magic angle spinning (HRMAS) NMR spectroscopic data of smoked Atlantic salmon (Salmo salar) were fully assigned by combination of one- and two-dimensional-HRMAS experiments. Complete representative spectra, obtained after few minutes of analysis time, revealed a large number of minor and major compounds in the sample. The methodology is limited by the low sensitivity of NMR, and therefore HRMAS only enables the determination of the most relevant components. These were fatty acids (FAs), carbohydrates, nucleoside derivatives, osmolytes, amino acids, dipeptides and organic acids. For the first time, spectra were resolved sufficiently to allow semiquantitative determination in intact muscle of the highly polyunsaturated FA 22:6 omega-3. Additionally, the feasibility of (1)H-HRMAS NMR metabolite profiling was tested to identify some bioactive compounds during storage. This profiling was carried out by the non-destructive and direct analysis (i.e. without requiring sample preparation and multiple step procedures) of intact salmon muscle. The proposed procedure can be applied to a large number of samples with high throughput due to the short time of analysis and quick evaluation of the data.