RESUMO
The first weeks of life represent a crucial stage for microbial colonization of the piglets' gastrointestinal tract. Newborns' microbiota is unstable and easily subject to changes under stimuli or insults. Nonetheless, the administration of antibiotics to the sow is still considered as common practice in intensive farming for pathological conditions in the postpartum. Therefore, transfer of antibiotic residues through milk may occurs, affecting the piglets' colon microbiota. In this study, we aimed to extend the knowledge on antibiotic transfer through milk, employing an in vitro dedicated piglet colon model (MICODE-Multi Unit In vitro Colon Model). The authors' focus was set on the shifts of the piglets' microbiota composition microbiomics (16S r-DNA MiSeq and qPCR-quantitative polymerase chain reaction) and on the production of microbial metabolites (SPME GC/MS-solid phase micro-extraction gas chromatography/mass spectrometry) in response to milk with different concentrations of amoxicillin. The results showed an effective influence of amoxicillin in piglets' microbiota and metabolites production; however, without altering the overall biodiversity. The scenario is that of a limitation of pathogens and opportunistic taxa, e.g., Staphylococcaceae and Enterobacteriaceae, but also a limitation of commensal dominant Lactobacillaceae, a reduction in commensal Ruminococcaceae and a depletion in beneficial Bifidobactericeae. Lastly, an incremental growth of resistant species, such as Enterococcaceae or Clostridiaceae, was observed. To the authors' knowledge, this study is the first evaluating the impact of antibiotic residues towards the piglets' colon microbiota in an in vitro model, opening the way to include such approach in a pipeline of experiments where a reduced number of animals for testing is employed. KEY POINTS: ⢠Piglet colon model to study antibiotic transfer through milk. ⢠MICODE resulted a robust and versatile in vitro gut model. ⢠Towards the "3Rs" Principles to replace, reduce and refine the use of animals used for scientific purposes (Directive 2010/63/UE).
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Microbioma Gastrointestinal , Microbiota , Animais , Suínos , Feminino , Amoxicilina/farmacologia , Colo , Metabolômica , Antibacterianos/farmacologiaRESUMO
Porcine models of ophthalmological diseases are often used in pre-clinical translational studies due to pigs' similarities to humans. In particular, the iodoacetic acid (IAA) model of photoreceptor degeneration seems to mimic well the endstage phenotype of human pathologies as retinitis pigmentosa and age-related macular degeneration, with high potential for prosthesis/retinal devices testing. IAA is capable of inducing photoreceptor death by blockage of glycolysis, and its effects on the retina have been described. Nonetheless, up to date, literature lacks of a comprehensive morpho-functional characterization of the entire visual system of this model. This gap is particularly critical for prosthesis testing as inner retinal structures and optic pathways must be preserved to elicit cortical responses and restore vision. In this study, we investigated the functional and anatomical features of the visual system of IAA-treated pigs and compared them to control animals. IAA was administered intravenously at 12 mg/kg; control animals received saline solution (NaCl 0.9% w/v). Electrophysiological analyses included full-field (ffERGs) and pattern (PERGs) electroretinograms and flash visually evoked potentials (fVEPs). Histological evaluations were performed on the retina and the optic pathways and included thickness of the different retinal layers, ganglion cells count, and immunohistochemistry for microglial cells, macroglial cells, and oligodendrocytes. The histological results indicate that IAA treatment does not affect the morphology of the inner retina and optic pathways. Electrophysiology confirms the selective rod and partial cone degeneration, but is ambiguous as to the functionality of the optic pathways, seemingly preserved as indicated by the still detectable fVEPs. Overall, the work ameliorates the characterization of such rapid and cost-effective model, providing more strength and reliability for future pre-clinical translational trials.
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Ácido Iodoacético/farmacologia , Células Fotorreceptoras Retinianas Cones/patologia , Degeneração Retiniana/tratamento farmacológico , Acuidade Visual , Animais , Modelos Animais de Doenças , Eletrorretinografia , Inibidores Enzimáticos/farmacologia , Feminino , Masculino , Reprodutibilidade dos Testes , Células Fotorreceptoras Retinianas Cones/metabolismo , Degeneração Retiniana/patologia , Degeneração Retiniana/fisiopatologia , SuínosRESUMO
The Raphe Pallidus (RPa) is a region of the brainstem that was shown to modulate the sympathetic outflow to many tissues and organs involved in thermoregulation and energy expenditure. In rodents, the pharmacological activation of RPa neurons was shown to increase the activity of the brown adipose tissue, heart rate, and expired CO2 , whereas their inhibition was shown to induce cutaneous vasodilation and a state of hypothermia that, when prolonged, leads to a state resembling torpor referred to as synthetic torpor. If translatable to humans, this synthetic torpor-inducing procedure would be advantageous in many clinical settings. A first step to explore such translatability, has been to verify whether the neurons within the RPa play the same role described for rodents in a larger mammal such as the pig. In the present study, we show that the physiological responses inducible by the pharmacological stimulation of RPa neurons are very similar to those observed in rodents. Injection of the GABAA agonist GABAzine in the RPa induced an increase in heart rate (from 99 to 174 bpm), systolic (from 87 to 170 mm Hg) and diastolic (from 51 to 98 mm Hg) arterial pressure, and end-tidal CO2 (from 49 to 62 mm Hg). All these changes were reversed by the injection in the same area of the GABAA agonist muscimol. These results support the possibility for RPa neurons to be a key target in the research for a safe and effective procedure for the induction of synthetic torpor in humans.
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Fármacos do Sistema Nervoso Autônomo/farmacologia , Neurônios/efeitos dos fármacos , Neurônios/fisiologia , Núcleo Pálido da Rafe/efeitos dos fármacos , Núcleo Pálido da Rafe/fisiologia , Fatores Etários , Animais , Feminino , Antagonistas GABAérgicos/administração & dosagem , Agonistas de Receptores de GABA-A/administração & dosagem , Frequência Cardíaca/efeitos dos fármacos , Frequência Cardíaca/fisiologia , Microinjeções/métodos , Piridazinas/administração & dosagem , Estremecimento/efeitos dos fármacos , Estremecimento/fisiologia , SuínosRESUMO
Antimicrobial resistance, an ever-growing global crisis, is strongly linked to the swine production industry. In previous studies, Melaleuca alternifolia and Rosmarinus officinalis essential oils have been evaluated for toxicity on porcine spermatozoa and for antimicrobial capabilities in artificial insemination doses, with the future perspective of their use as antibiotic alternatives. The aim of the present research was to develop and validate in vitro and ex vivo models of porcine uterine mucosa for the evaluation of mucosal toxicity of essential oils. The in vitro model assessed the toxicity of a wider range of concentrations of both essential oils (from 0.2 to 500 mg/mL) on sections of uterine tissue, while the ex vivo model was achieved by filling the uterine horns. The damage induced by the oils was assessed by Evans Blue (EB) permeability assay and histologically. The expression of ZO-1, a protein involved in the composition of tight junctions, was assessed through immunohistochemical and immunofluorescence analysis. The results showed that low concentrations (0.2-0.4 mg/mL) of both essential oils, already identified as non-spermicidal but still antimicrobial, did not alter the structure and permeability of the swine uterine mucosa. Overall, these findings strengthen the hypothesis of a safe use of essential oils in inseminating doses of boar to replace antibiotics.
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Anti-Infecciosos/toxicidade , Melaleuca/química , Mucosa/efeitos dos fármacos , Óleos Voláteis/toxicidade , Rosmarinus/química , Óleo de Melaleuca/toxicidade , Útero/efeitos dos fármacos , Animais , Anti-Infecciosos/farmacologia , Corantes/farmacocinética , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Epitélio/efeitos dos fármacos , Epitélio/ultraestrutura , Azul Evans/farmacocinética , Feminino , Inseminação Artificial/veterinária , Masculino , Microscopia de Fluorescência , Óleos Voláteis/farmacologia , Permeabilidade/efeitos dos fármacos , Preservação do Sêmen/métodos , Espermatozoides/efeitos dos fármacos , Suínos , Óleo de Melaleuca/farmacologia , Junções Íntimas/efeitos dos fármacos , Útero/ultraestrutura , Proteína da Zônula de Oclusão-1/análiseRESUMO
The growing interest towards essential oils stems from their biological capabilities that include antibacterial and antioxidant effects. Such properties may be extremely useful in the reproductive field; nonetheless essential oils show toxic effects that can lead to cell disruption. The present study aimed to evaluate and compare the effects of tea tree oil (TTO) and its principal component terpinen-4-ol (TER) on the morpho-functional parameters of swine spermatozoa. Experimental samples were prepared by suspending 15 × 107 spermatozoa in 5 mL of medium with different concentrations of the above-mentioned compounds: from 0.2 to 2 mg/mL at an interval of 0.2 for TTO, while TER concentrations were adjusted according to its presence in TTO (41.5%). After 3 h incubation at 16 °C, samples were analyzed for pH, viability, acrosome status, and objective motility. The results highlighted a concentration-dependent effect of TTO with total motility as the most sensitive parameter. TER was better tolerated, and the most sensitive parameters were related to membrane integrity, suggesting a different pattern of interaction. The study confirms the importance of evaluating the effects of natural compounds on spermatozoa before exploiting their beneficial effects. Spermatozoa seem to be good candidates for preliminary toxicological screenings in the light of their peculiar properties.
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Melaleuca/química , Espermatozoides/efeitos dos fármacos , Óleo de Melaleuca/farmacologia , Terpenos/farmacologia , Animais , Masculino , Suínos , Terpenos/químicaRESUMO
BACKGROUND: The 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. RESULTS: Significant 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. CONCLUSIONS: In 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.
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Envelhecimento/sangue , Contagem de Células Sanguíneas/veterinária , Hemoglobinas/metabolismo , Compostos de Ferro/farmacologia , Suínos/sangue , Envelhecimento/fisiologia , Animais , Análise Química do Sangue/veterinária , Glicemia , Eletrólitos/sangue , Feminino , Hematócrito/veterinária , Testes Hematológicos/veterinária , Injeções , Compostos de Ferro/sangue , Masculino , Valores de Referência , Suínos/fisiologia , Oligoelementos/sangueRESUMO
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.
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Óleos Voláteis/química , Óleos Voláteis/farmacologia , Rosmarinus/química , Espermatozoides/efeitos dos fármacos , Animais , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Gasosa-Espectrometria de Massas , Masculino , Óleos de Plantas/química , Óleos de Plantas/farmacologia , Espermatozoides/ultraestrutura , SuínosRESUMO
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.
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Raquianestesia/veterinária , Cateterismo/veterinária , Medula Espinal , Ultrassonografia de Intervenção/veterinária , Raquianestesia/métodos , Animais , Animais Recém-Nascidos , Cateterismo/métodos , Cateteres de Demora/veterinária , Vértebras Lombares/diagnóstico por imagem , Medula Espinal/diagnóstico por imagem , Suínos , Vértebras Torácicas/diagnóstico por imagem , Ultrassonografia de Intervenção/métodosRESUMO
The use of phytoextracts has been proposed as a method to improve animal welfare, also in pigs, by reducing stress and anxiety and improving performances. Lavandula angustifolia (Miller) essential oil (LaEO) is an interesting calming phytoextract that could be administered by inhalation for prolonged periods of time to help pigs coping with on-farm conditions. The aim of this study was to assess the effects of daily inhalation of vaporized LaEO on pigs' welfare and health indicators, and nasal microbiota, trying to understand whether this phytoextract represents a feasible tool to improve animal welfare under intensive farming conditions. Eighty-four crossbred barrows were randomly divided into 3 experimental groups: control (C); lavender (L): 3 vaporization sessions of 10 min each of a custom made 1% solution of LaEO; sham (S): same vaporization sessions of L group but only using the solution vehicle. Experimental readouts included growth parameters, behavioural traits, tail and skin lesions, hair steroids and nasal microbiota. L group animals did not show altered growth performance and seemed calmer (increased recumbency time), with decreased amount of skin lesions also associated with lower severity class for tail lesions. They also showed decreased CORT/DHEA ratio, potentially suggesting a beneficial effect of LaEO. Inhalation of LaEO significantly affected the nasal pig microbiome by reducing its diversity. Overall, the study suggests how inhalation of Lavender essential oil may be capable of improving welfare in growing pigs, yet it is pivotal to consider the microbial modulatory capabilities of essential oils before exploiting them on larger scale.
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Lavandula , Óleos Voláteis , Animais , Suínos , Óleos Voláteis/farmacologia , Óleos de Plantas/farmacologiaRESUMO
The value of pig as "large animal model" is a well-known tool for translational medicine, but it can also be beneficial in studying animal health in a one-health vision. The ConcePTION Project aims to provide new information about the risks associated with medication use during breastfeeding, as this information is not available for most commonly used drugs. In the IMI-Conception context, Göttingen Minipigs have been preferred to hybrid pigs for their genetic stability and microbiological control. For the first time, in the present research, three primary cell cultures of mammary epithelial cells were isolated and characterized from Göttingen Minipigs (mpMECs), including their ability to create the epithelial barrier. In addition, a comparative analysis between Göttingen Minipigs and commercial hybrid pig mammary epithelial cells (pMECs) was conducted. Epithelial markers: CKs, CK18, E-CAD, ZO-1 and OCL, were expressed in both mpMECs and pMECs. RT2 Profiler PCR Array Pig Drug Transporters showed a similar profile in mRNA drug transporters. No difference in energy production under basal metabolic condition was evidenced, while under stressed state, a different metabolic behaviour was shown between mpMECs vs pMECs. TEER measurement and sodium fluorescein transport, indicated that mpMECs were able to create an epithelial barrier, although, this turned out to be less compact than pMECs. By comparing mpMECs with mammary epithelial cells isolated from Hybrid pigs (pMECs), although both cell lines have morphological and phenotypic characteristics that make them both useful in barrier studies, some specific differences exist and must be considered in a translational perspective.
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Células Epiteliais , Glândulas Mamárias Animais , Porco Miniatura , Animais , Suínos , Feminino , Glândulas Mamárias Animais/citologia , Células CultivadasRESUMO
The application of essential oils as potential alternatives to antibiotics in swine semen storage is promising, due to their antioxidant and antibacterial properties. However, detrimental effects on spermatozoa should be clarified first. The aim of this study was to evaluate 9 essential oils (EOs; Satureja montana, Pelargonium graveolens, Cymbopogon nardus, Melaleuca leucadendron, Eucaliptus globulus, Citrus limon, Lavandula angustifolia, Lavandula hybrida, Mentha piperita) and a blend (GL mix) on key morpho-functional parameters of swine spermatozoa. Test compounds were firstly chemo-characterized and experimental doses were prepared by suspending a fixed number of spermatozoa with 3 different concentrations (0.1, 0.5, 1 mg/mL) of EOs. Experimental doses were stored at 16 °C and sampled after 3 and 120 h for analysis. Overall, S. montana, P. graveolens and L. angustifolia EOs induced the strongest alterations, with C. nardus and E. globulus EOs being the best tolerated. Swine spermatozoa represent a good preliminary testing platform to screen toxicity and its different patterns. The comprehensive overview on the potential mechanisms of action of some of the most common EOs, despite of the direct aim of the study being swine reproduction, may be exploited in other fields of research within both veterinary and human medicine.
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Lavandula , Óleos Voláteis , Masculino , Suínos , Humanos , Animais , Óleos Voláteis/farmacologia , Refrigeração , Espermatozoides , Antibacterianos/farmacologiaRESUMO
INTRODUCTION: Insufficient supply of cardiac grafts represents a severe obstacle in heart transplantation. Donation after Circulatory Death (DCD), in addition to conventional donation after brain death, is one promising option to overcome the organ shortage. However, DCD organs undergo an inevitable more extended period of warm unprotected ischemia between circulatory arrest and graft procurement. Mesenchymal stromal cell-derived extracellular vesicles (MSC-EVs) have shown remarkable protective effects against ischemia-reperfusion injury. Thus, we aimed to enhance grafts preservation from DCD donors, through treatment with MSC-EVs. METHODS: Female pigs were euthanized by barbiturate overdose and after 20â¯min of a flat EKG, the chest was opened, the heart harvested and subsequently connected to an extracorporeal perfusion machine. MSC-EVs, isolated by ion exchange chromatography, were added to the perfusion solution (1×1011 particles) and the heart was perfused for 2â¯h. Then, heart tissue biopsies were taken to assess histological changes, mitochondrial morphology, antioxidant enzyme activity and inflammation mediators' expression. Biochemical parameters of myocardial viability were assessed in the perfusate. RESULTS: The treatment with MSC-EVs significantly prevented mitochondria swelling, mitochondrial cristae loss and oxidative stress in cardiac tissue. The protective effect of MSC-EVs was confirmed by the delayed increase of the cardiac-specific enzymes CK and TnC in the perfusate and the reduction of caspase-3+ cells in tissue sections. CONCLUSION: MSC-EVs improve graft quality by preserving the mitochondrial ultrastructure protecting the myocardium against oxidative stress, reducing apoptosis of cardiac cells and preventing the increase of pro-inflammatory cytokines.
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Vesículas Extracelulares , Células-Tronco Mesenquimais , Animais , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/transplante , Células-Tronco Mesenquimais/metabolismo , Feminino , Suínos , Estresse Oxidativo , Transplante de Coração/métodos , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Traumatismo por Reperfusão Miocárdica/patologia , Modelos Animais de Doenças , Miocárdio/patologia , Miocárdio/metabolismo , Traumatismo por Reperfusão/prevenção & controle , Traumatismo por Reperfusão/patologiaRESUMO
Electrocorticography is an established neural interfacing technique wherein an array of electrodes enables large-area recording from the cortical surface. Electrocorticography is commonly used for seizure mapping however the implantation of large-area electrocorticography arrays is a highly invasive procedure, requiring a craniotomy larger than the implant area to place the device. In this work, flexible thin-film electrode arrays are combined with concepts from soft robotics, to realize a large-area electrocorticography device that can change shape via integrated fluidic actuators. We show that the 32-electrode device can be packaged using origami-inspired folding into a compressed state and implanted through a small burr-hole craniotomy, then expanded on the surface of the brain for large-area cortical coverage. The implantation, expansion, and recording functionality of the device is confirmed in-vitro and in porcine in-vivo models. The integration of shape actuation into neural implants provides a clinically viable pathway to realize large-area neural interfaces via minimally invasive surgical techniques.
Assuntos
Eletrocorticografia , Eletrodos Implantados , Eletrocorticografia/instrumentação , Eletrocorticografia/métodos , Animais , Suínos , Craniotomia/métodos , Craniotomia/instrumentação , Procedimentos Cirúrgicos Minimamente Invasivos/instrumentação , Procedimentos Cirúrgicos Minimamente Invasivos/métodos , Robótica/instrumentação , Robótica/métodos , Encéfalo/fisiologiaRESUMO
Two main classes of perivascular multipotent populations have been described: the microvascular pericytes and the vascular wall mesenchymal stem cells (VW-MSCs). VW-MSCs are isolated from large vessels in many species and they participate in vascular remodeling together with other cellular components such as endothelial cells. Considering that the Göttingen Minipigs are widely used in Europe as a translational model in the field of cardiovascular diseases, the aim of the present research was to isolate VW-MSCs from the adult aorta of Göttingen Minipigs while preserving and also collecting endothelial cells. The results obtained in the present research demonstrated that this new protocol allows us to obtain a pure population of VW-MSCs and endothelial cells. VW-MSCs from Göttingen Minipigs responded fully to the MSC minima international criteria, being positive to CD105, CD90, and CD44 and negative to CD45 and CD34. Moreover, VW-MSCs presented a differentiative potential towards osteogenic, chondrogenic, and adipogenic lineages. Overall, the present protocol, preserving the viability and phenotypic features of the two isolated populations, opens future possibilities of using minipig VW-MSCs and endothelial cells in in vitro vascular remodeling studies.
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INTRODUCTION: We have recently developed a new miniaturized device for extracorporeal ultrafiltration (UF) to be used in patients with fluid overload: Artificial Diuresis-1 (AD1) (Medica S.p.A., Medolla, Italy). The device has a reduced priming volume, operates at very low pressures and flow regimes, and is designed to perform extracorporeal UF at bedside. After accurate experiments were carried out in vitro, we report in this paper the results of in vivo UF sessions carried out in selected animals according to veterinary best practice. MATERIALS AND METHODS: The AD1 kit is pre-filled with sterile isotonic solution and operates with a polysulfone mini-filter, MediSulfone (polysulfone at 50,000 Dalton). A collection bag with a volumetric scale is connected to the UF line, and the ultrafiltrate is obtained by gravity based on the height at which the ultrafiltrate collection bag is placed. Animals were prepared and anesthetized. The jugular vein was cannulated with a double-lumen catheter. Three 6-h sessions of UF were scheduled with a target fluid removal of 1,500 mL. Heparin was used as anticoagulant. RESULTS: In all treatments, the target value of UF was obtained in the absence of major clinical or technical problems with a maximum deviation from the scheduled UF rate lower than 10%. The device resulted to be safe, reliable, accurate, and easily usable thanks to a user-friendly interface and its very small dimensions. CONCLUSIONS: This study opens the way for clinical trials in different settings including departments with low intensity of care and even in ambulatory centers or patient's home.
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Diurese , Ultrafiltração , Humanos , Animais , Ultrafiltração/métodos , Anticoagulantes , ItáliaRESUMO
The European eel is a critically endangered teleost fish with very poor success rate for captive breeding and artificial reproduction. Therefore, to support its conservation, new strategies are needed to ensure fertilization. Objective analysis of sperm motility may be critical as it potentially represents one of the most important reproductive quality parameters. Spermatozoa acquire motility once in contact with hyperosmotic solutions as saltwater, yet the exact mechanisms and the role of temperature are still to be clarified. The main aim of the study was to assess the effects of 3 activating media (artificial sea water, tank water and commercial Actifish®) at 4 and 20 °C on sperm motility, by means of computer assisted sperm analysis. Secondary aim was to test 2 different concentrations of Actifish® mimicking sea water pH/osmolality, at 4 °C. The results suggested how both temperature and activating media have effects on spermatozoa motility and kinematics, with temperature mainly acting upon interaction with the media type. The samples activated with tank water at 20 °C showed the poorest motility outcomes (mean 38.1%), while the ones activated with Actifish® diluted 1:4 and artificial sea water, at 4 °C, the highest (means 51.8 and 51.5% respectively). Additionally, diluting Actifish® to reach same pH and osmolality of seawater led to worse motility outcomes, suggesting that composition may be the critical factor for activation rather than osmolality itself.
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Anguilla , Motilidade dos Espermatozoides , Masculino , Animais , Temperatura , Sêmen , Espermatozoides , Água , ComputadoresRESUMO
Octopus vulgaris is a commercially valuable species. It is overexploited in the natural environment and is considered to be an innovative species for aquaculture. However, large-scale farming is generally designed only based on economic requirements, disregarding any form of enrichment that induces the natural behavior of aquatic species. Although many studies have shown the influence of environmental enrichment on terrestrial vertebrates, fish, and cephalopod mollusks, information on the effect of environmental enrichment on the body patterns of O. vulgaris is limited. Therefore, in this study, we assessed how different environmental conditions (Basic vs. Enriched) affect sub-adults of O. vulgaris kept in recirculation systems, through qualitative-quantitative studies of the main body patterns and their potential application in the commercial production of this species. The results indicated that octopuses kept in the enriched environment showed several body patterns and gained a significantly higher weight than those kept in the basic environment. The body patterns displayed by the individuals kept in the basic environment were similar to those exhibited under situations of hostility and inter/intra-specific conflict. Hence, the environment of octopuses needs to be enriched, especially for the large-scale production of this species.
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Background and Aim: Seasonal changes, especially temperature and photoperiod, are well-known determining factors of swine reproductive capacity, but the underlying mechanisms remain unknown. This study aimed to investigate the effect of age and seasonal variations on boar seminal plasma steroids (dehydroepiandrosterone [DHEA], cortisol [CORT], and testosterone [TEST]) over 1 year. Materials and Methods: Four commercial hybrid adult boars (Large White × Duroc), aged between 12 and 44 months, were repeatedly evaluated at the Department of Veterinary Medical Sciences of the University of Bologna. Daily temperature and light hours relating to the collection date were considered for each observation within the four astronomical seasons: Winter, spring, summer, and autumn. Hormones were quantified using radioimmunoassay. The association between seasonal factors and hormone concentrations was evaluated using linear regression models. Univariate models were estimated for each hormone to assess the influence of the independent variables; two multivariate models were assessed to evaluate the effect of temperature and daylight hours, including boar and season factors. Results: Age significantly affected all analyzed hormones (CORT p < 0.0001; DHEA p < 0.0001; and TEST p < 0.0001). The highest average levels were found for each hormone during summertime, suggesting a positive correlation between steroid concentrations with temperature and light hours. Conclusion: The results of this study support the hypothesis that the increase in external temperature and light hours is somehow associated with higher levels of steroid concentrations in the seminal plasma of in-housed boars. These findings may help further investigate seasonal fluctuations in reproductive outcomes, which are well-known for porcine species.
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Pigs are becoming an important pre-clinical animal species for translational ophthalmology, due to similarities with humans in anatomical and physiological patterns. Different models of eye disorders have been proposed, and they are good candidates to assess biocompatibility/functionality of retinal prostheses. Electroretinography is a common tool allowing to gain information on retinal function, with several types of electroretinogram (ERG) been implemented including full field (ff-ERG), multifocal (mf-ERG) and pattern (p-ERG). p-ERG represents a valuable tool to monitor Retinal Ganglion Cells (RGCs) activity and can be used to calculate p-ERG spatial acuity. Unfortunately, scarce methodological data are available regarding recording/interpretation of p-ERG and retinal acuity in biomedical pigs yet enhancing knowledge regarding pig vision physiology will allow for more refined and responsible use of such species. Aim of this study was to record p-ERG in juvenile pigs to functionally assess visual acuity. Six female hybrid pigs underwent two p-ERG recording sessions at 16 and 19 weeks of age. Photopic ff-ERG were also recorded; optical coherence tomography (OCT) and histology were used to confirm retinal integrity. ff-ERG signals were repeatable within/across sessions. All p-ERG traces consistently displayed characterizing peaks, and the progressive decrease of amplitude in response to the increment of spatial frequency revealed the reliability of the method. Mean p-ERG spatial acuities were 5.7 ± 0.14 (16 weeks) and 6.2 ± 0.15 cpd (19 weeks). Overall, the p-ERG recordings described in the present work seem reliable and repeatable, and may represent an important tool when it comes to vision assessment in pigs.
Assuntos
Eletrorretinografia , Doenças Retinianas , Animais , Eletrorretinografia/métodos , Feminino , Humanos , Reprodutibilidade dos Testes , Retina/patologia , Doenças Retinianas/patologia , Suínos , Tomografia de Coerência Óptica , Regulador Transcricional ERGRESUMO
The extensive use of antibiotics has contributed to the current antibiotic resistance crisis. Livestock infections of Salmonella spp, Clostridium spp. and E. coli antimicrobial-resistant bacteria represent a public threat to human and animal health. To reduce the incidence of these zoonoses, essential oils (EOs) could be effective antibiotic alternatives. This study aims at identifying EOs safe for use, effective both in complementary therapy and in the environmental sanitization of intensive farming. Natural products were chemo-characterized by gas chromatography. Three S. Typhimurium, three C. perfringens and four E. coli strains isolated from poultry and swine farms were used to assess the antimicrobial properties of nine EOs and a modified GR-OLI (mGR-OLI). The toxicity of the most effective ones (Cinnamomum zeylanicum, Cz; Origanum vulgare, Ov) was also evaluated on porcine spermatozoa and Galleria mellonella larvae. Cz, Ov and mGR-OLI showed the strongest antimicrobial activity; their volatile components were also able to significantly inhibit the growth of tested strains. In vitro, Ov toxicity was slightly lower than Cz, while it showed no toxicity on G. mellonella larvae. In conclusion, the study confirms the importance of evaluating natural products to consolidate the idea of safe EO applications in reducing and preventing intensive livestock infections.