Alberto Elmi

Age-Related 1H NMR Characterization of Cerebrospinal Fluid in Newborn and Young Healthy Piglets.

When it comes to neuroscience, pigs represent an important animal model due to their resemblance with humans’ brains for several patterns including anatomy and developmental stages. Cerebrospinal fluid (CSF) is a relatively easy-to-collect specimen that can provide important information about neurological health and function, proving its importance as both a diagnostic and biomedical monitoring tool. Consequently, it would be of high scientific interest and value to obtain more standard physiological information regarding its composition and dynamics for both swine pathology and the refinement of experimental protocols. Recently, proton nuclear magnetic resonance (1H NMR) spectroscopy has been applied in order to analyze the metabolomic profile of this biological fluid, and results showed the technique to be highly reproducible and reliable. The aim of the present study was to investigate in both qualitative and quantitative manner the composition of Cerebrospinal Fluid harvested form healthy newborn (5 days old-P5) and young (30-P30 and 50-P50 days old) piglets using 1H NMR Spectroscopy, and to analyze any possible difference in metabolites concentration between age groups, related to age and Blood-Brain-Barrier maturation. On each of the analyzed samples, 30 molecules could be observed above their limit of quantification, accounting for 95–98% of the total area of the spectra. The concentrations of adenine, tyrosine, leucine, valine, 3-hydroxyvalerate, 3-methyl-2-oxovalerate were found to decrease between P05 and P50, while the concentrations of glutamine, creatinine, methanol, trimethylamine and myo-inositol were found to increase. The P05-P30 comparison was also significant for glutamine, creatinine, adenine, tyrosine, leucine, valine, 3-hydroxyisovalerate, 3-methyl-2-oxovalerate, while for the P30-P50 comparison we found significant differences for glutamine, myo-inositol, leucine and trimethylamine. None of these molecules showed at P30 concentrations outside the P05 –P50 range.

The biomedical piglet: establishing reference intervals for haematology and clinical chemistry parameters of two age groups with and without iron supplementation

BackgroundThe similarities between swine and humans in physiological and genomic patterns, and the great correlation in size and anatomy, make pigs extremely useful in preclinical studies. New-born piglets can represent a model for congenital and genetic diseases in new-born children. It is known that piglets may have significant differences in clinicopathological results compared to adult pigs. Therefore, adult laboratory reference intervals cannot be applied to piglets. The aim of this study was to compare haematological and chemical variables in piglets of two ages and determinate age-related reference intervals for commercial hybrid young pigs.Blood samples were collected under general anaesthesia from 130 animals divided into five- (P5) and 30- (P30) day-old piglets. Only P30 animals were treated with parenteral iron after birth. Samples were analysed using automated haematology (ADVIA 2120) and chemistry analysers, and age-related reference intervals were calculated.ResultsSignificant higher values of RBC, Hb and HCT were observed in P30 animals when compared to P5, with an opposite trend for MCV. These results were associated with a reduction of the RBC regeneration process and the thrombopoietic response. The TSAT and TIBC were significantly higher in P30 compared to P5; however, piglets remained iron deficient compared to adult reference intervals reported previously.ConclusionsIn conclusion, this paper emphasises the high variability occurring in clinicopathological variables between new-born and 30-day-old pigs, and between piglets and adult pigs. This study provides valuable reference data for piglets at precise ages and could be used in the future as historical control improving the Reduction in animal experiments, as suggested by the 3Rs principle.

Thymbra capitata (L.) Cav. and Rosmarinus officinalis (L.) Essential Oils: In Vitro Effects and Toxicity on Swine Spermatozoa

Essential oils possess a variety of biological properties (i.e., antioxidant, antibacterial, and cytotoxic) that could possibly be applied in reproductive medicine, but their effects on spermatozoa are still partially unknown. The aim of the study was to describe the effects of Thymbra capitata (L.) Cav. and Rosmarinus officinalis (L.) essential oils on the main morpho-functional parameters of swine spermatozoa. Essential oils were preliminary characterized by gas chromatography and added with emulsifiers to facilitate diffusion. Experimental samples were prepared by suspending a fixed number of spermatozoa in 5 mL of medium with 10 different concentrations of essential oil (0.2–2 mg/mL, at intervals of 0.2). After 3 h of incubation, samples were analyzed for pH, viability, objective motility, and acrosome status. Results showed that the effects of the essential oils are concentration-dependent and that R. officinalis is well tolerated up to 0.6 mg/mL. T. capitata impaired the spermatozoa starting from the lowest concentration, with complete spermicidal effect from 0.4 mg/mL. The patterns of damage, confirmed by SEM, were different and quite distinct. As expected, spermatozoa proved to be sensitive to external stimuli and capable of showing different functional patterns, providing interesting insights to the action/toxicity mechanisms. The results of the present work represent the first step towards the systematic characterization of the effects of these compounds on spermatozoa. This kind of studies are necessary to strengthen the idea of future applications of essential oils in the reproductive field due to their antioxidant, antibacterial, or spermicidal properties.

Semen evaluation and in vivo fertility in a Northern Italian pig farm: Can advanced statistical approaches compensate for low sample size? An observational study

The evaluation of sperm functionality and morphology allows discerning between high and low quality ejaculates, but does not give detailed predictive information regarding in vivo fertility. The current developments in statistical modeling have helped in carrying out reproductive studies, but their biggest limitation is in the size of the dataset to be used. The aim of the present observational study was to evaluate whether advanced statistical approaches, such as mixed effects regression models and bootstrap resampling, can help in assessing the predictive ability of semen parameters in terms of in vivo fertility (farrowing rate and litter size), on a small/medium farm with a limited number of animals. Data regarding 33 ejaculates, including viability, subjective motility and acrosome reaction, were collected. Two hundred and thirty-five sows were inseminated with an outcome of 167 deliveries and 1734 newborn piglets. In order to evaluate the relationships among the parameters measured and fertility, mixed effects regression statistical models were used. Once the covariates to be included in the final models were identified, non-parametric bootstrapping was used. The results showed that the farrowing rate was highly associated with the total number of spermatozoa and subjective motility, while litter size was associated with percentage of acrosome reaction. In conclusion, the proposed statistical approach seemed to be suitable for studies regarding reproduction and fertility, even for relatively small sample sizes. Nonetheless, larger data sets are still preferable and required in order to achieve higher reliability.

Hair Testosterone and Cortisol Concentrations in Pre- and Post-Rut Roe Deer Bucks: Correlations with Blood Levels and Testicular Morphometric Parameters

Simple Summary The roe deer is a very common wild species in Italy and shows peculiar reproductive characteristics. Sexually-mature males, called bucks, show a complete interruption in spermatogenesis during the cold seasons. The mechanisms behind such interruption are still partially unknown. Hair is a good biological sample, easy to obtain while minimizing stress, for endocrinological analyses that may provide information regarding such mechanisms. The aim of the work was to quantify and compare hair concentrations of testosterone and cortisol in wild roe deer bucks hunted during the pre- and post-rut period (mating season). The secondary objective was the evaluation of possible correlations of such hair concentrations with blood and morphometric parameters of the testes. Testosterone significantly increased from the pre- to post-rut period, while cortisol significantly decreased. The correlations with blood and testicular parameters resemble what is already described in the literature. Overall, this study represents a first report of the quantification of testosterone and cortisol in roe deer hair, and may provide interesting insights into their reproductive physiology. Abstract The roe deer is a seasonally breeding species with a reproductive cycle regulated by endogenous rhythms and photoperiod-sensitivity. Sexually mature bucks show hormonal and testicular activation during the reproductive season, with a peak in the rut period, and following gradual involution. Hair is a good matrix for non-invasive endocrinological analyses that provide long-term information without being influenced by the hormones’ pulsating release patterns in blood. The aim of the work was to quantify hair concentrations of testosterone and cortisol in wild roe deer bucks hunted during the pre- and post-rut period, using a radioimmunoassay methodology, and to look for differences between the two periods. The secondary objective was the evaluation of possible correlations of such hair concentrations with blood and morphometric parameters of the testes. Both hormones showed statistical differences, with opposing trends, when comparing the two periods: testosterone increased while cortisol decreased. The correlation analysis was in agreement with existing literature regarding metabolism/actions of these hormones and testicular morphometric parameters. This study represents the first report of the use of radioimmunoassay techniques to quantify testosterone and cortisol in roe deer hair, and may provide interesting insights into their reproductive physiology.

Ultrasound guided spinal catheter insertion in piglet: preliminary results

OBJECTIVE To describe the ultrasound (US) evaluation of the cervical, thoracic and lumbar spinal tracts in piglets and to evaluate the feasibility of the ultrasound guided spinal catheter placement in newborn and paediatric piglets. STUDY DESIGN Prospective experimental study. ANIMALS A total of two piglet cadavers (age, 7 and 14 days) and eight commercial crossbreed piglets divided into four groups according to age: 7 (P7), 14 (P14), 21 (P21) and 28 (P28) days. METHODS In the first part of the study an ultrasound examination of the spinal tract was performed in piglet cadavers applying the transverse and the longitudinal approaches in sternal and lateral recumbencies. In the second phase, the piglets were anaesthetized with sevoflurane. A US examination with a 10 MHz linear probe was performed and a spinal catheter was introduced between the spinous processes of L2 and L3 lumbar vertebrae using an in-plane technique and its advancement was monitored with the probe. At the end of procedure, the catheter was removed. The piglets recovered from anaesthesia and were monitored for one week. RESULTS In phase I the authors identified the paramedian longitudinal approach as the most feasible for spinal structure evaluation in piglets. In phase II, the paramedian longitudinal views enabled a good visualization of the spinal cord and of the catheter advancement up to the cisterna magna in groups P7 and P14. In groups P21 and P28 it was not possible to visualize the neuroaxial structures and the spinal catheter using the same approach. No clinical alterations were recorded during the procedure or the following days. CONCLUSIONS AND CLINICAL RELEVANCE US-guided spinal catheter placement appeared a feasible technique in piglets younger than 14 days but it is not useful in older piglets.