Recent studies performed in humans and rats have reported that exercise can alter the intestinal microbiota. Athletic horses perform intense exercise regularly, but studies characterizing horse microbiome during aerobic conditioning programs are still limited. Evidence has indicated that this microbial community is involved in the metabolic homeostasis of the host. Research on ergogenic substances using new sequencing technologies have been limited to the intestinal microbiota and there is a considerable demand for scientific studies that verify the effectiveness of these supplements in horses. L-carnitine and chromium are potentially ergogenic substances for athletic humans and horses since they are possibly able to modify the metabolism of carbohydrates and lipids. This study aimed to assess the impact of acute exercise and aerobic conditioning, associated either with L-carnitine or chromium supplementation, on the intestinal microbiota of fillies. Twelve “Mangalarga Marchador” fillies in the incipient fitness stage were distributed into four groups: control (no exercise), exercise, L-carnitine (10g/day) and chelated chromium (10mg/day). In order to investigate the impact of acute exercise or aerobic conditioning on fecal microbiota all fillies undergoing the conditioning program were analyzed as a separate treatment. The fillies underwent two incremental exercise tests before and after training on a treadmill for 42 days at 70–80% of the lactate threshold intensity. Fecal samples were obtained before and 48 h after acute exercise (incremental exercise test). Bacterial populations were characterized by sequencing the V4 region of the 16S rRNA gene using the MiSeq Illumina platform, and 5,224,389 sequences were obtained from 48 samples. The results showed that, overall, the two most abundant phyla were Firmicutes (50.22%) followed by Verrucomicrobia (15.13%). The taxa with the highest relative abundances were unclassified Clostridiales (17.06%) and "5 genus incertae sedis" from the phylum Verrucomicrobia (12.98%). There was a decrease in the phylum Chlamydiae and in the genus Mycobacterium after the second incremental exercise test. Intense exercise changed the community’s structure and aerobic conditioning was associated with changes in the composition and structure of the intestinal bacterial population of fillies. The intra-group comparison showed that chromium or L-carnitine induced moderate changes in the fecal microbiota of fillies, but the microbiota did not differ from the control group, which was exercised with no supplementation. Fecal pH correlated positively with Simpson’s index, while plasma pH correlated negatively. Our results show that exercise and aerobic conditioning can change in the microbiota and provide a basis for further studies enrolling a larger number of horses at different fitness levels to better understand the effects of exercise and training on the intestinal microbiota of horses.
Poly(lactic acid) (PLA) and poly(ε-caprolactone) (PCL) are two important aliphatic esters known for their biodegradability and bioresorbability properties; the former is stiffer and brittle while the smaller modulus of the latter allows a suitable elongation. The new biomaterials being developed from the blend of these two polymers (PLA and PCL) is opportune due to the reducing interfacial tension between their immiscible phases. In a previous study, PLA/PCL immiscible blend when compatibilized with poly(ε-caprolactone-b-tetrahydrofuran) resulted in enhanced ductility and toughness no cytotoxic effect in vitro tests. There is little published data on the effect of poly(ε-caprolactone-b-tetrahydrofuran) on PLA and PCL biocompatibility and biodegradability in vivo tests. This study focuses on evaluating the behavioral response and polymer-tissue interaction of compatibilized PLA/PCL blend compared to neat PLA implanted via intraperitoneal (IP) and subcutaneous (SC) in male Wistar rats, distributed in four experimental groups: neat PLA, PLA/PCL blend, sham, and control at 2-, 8- and 24-weeks post-implantation (WPI). An open-field test was performed to appraise emotionality and spontaneous locomotor activity. Histopathological investigation using hematoxylin-eosin (H&E) and picrosirius-hematoxylin (PSH) was used to assess polymer-tissue interaction. Modifications in PLA and the PLA/PCL blend’s surface morphology were determined by scanning electron microscopy (SEM). PLA group defecated more often than PLA/PCL rats 2 and 8 WPI. Conjunctive capsule development around implants, cell adhesion, angiogenesis, and giant cells of a foreign body to the biomaterial was observed in light microscopy. Both groups displayed a fibrous reaction along with collagen deposition around the biomaterials. In the SEM, the images showed a higher degradation rate for the PLA/PCL blend in both implantation routes. The polymers implanted via IP exhibited a higher degradation rate compared to SC. These findings emphasize the biocompatibility of the PLA/PCL blend compatibilized with poly(ε-caprolactone-b-tetrahydrofuran), making this biopolymer an acceptable alternative in a variety of biomedical applications.
In horses, there is an increasing interest in developing long-lasting drug formulations, with biopolymers as viable carrier alternatives in addition to their use as scaffolds, suture threads, screws, pins, and plates for orthopedic surgeries. This communication focuses on the prolonged biocompatibility and biodegradation of PLA, prepared by hot pressing at 180 °C. Six samples were implanted subcutaneously on the lateral surface of the neck of one horse. The polymers remained implanted for 24 to 57 weeks. Physical examination, plasma fibrinogen, and the mechanical nociceptive threshold (MNT) were performed. After 24, 28, 34, 38, and 57 weeks, the materials were removed for histochemical analysis using hematoxylin-eosin and scanning electron microscopy (SEM). There were no essential clinical changes. MNT decreased after the implantation procedure, returning to normal after 48 h. A foreign body response was observed by histopathologic evaluation up to 38 weeks. At 57 weeks, no polymer or fibrotic capsules were identified. SEM showed surface roughness suggesting a biodegradation process, with an increase in the median pore diameter. As in the histopathological evaluation, it was not possible to detect the polymer 57 weeks after implantation. PLA showed biocompatible degradation and these findings may contribute to future research in the biomedical area.
Facial-expression-based analysis has been widely applied as a pain coding system in horses. Herein, we aimed to identify pain in horses undergoing subcutaneously polylactide-based polymer implantation. The sham group was submitted only to surgical incision. The horses were filmed before and 24 and 48 h after implantation. Five statistical methods for evaluating their facial expressions (FEs) were tested. Primarily, three levels of scores (0, 1, and 2) were applied to the seven FEs (ear movements, eyebrow tension, orbicularis tension, dilated nostrils, eye opening, muzzle tension, and masticatory muscles tension). Subsequently, the scores of the seven FEs were added (SUM). Afterwards, principal component analysis (PCoA) was performed using the scores of the seven FEs obtained using the first method. Subsequently, weights were created for each FE, based on each variable’s contribution variability obtained from the PCoA (SUM.W). Lastly, we applied a general score (GFS) to the animal’s face (0 = without pain; 1 = moderate pain; 2 = severe pain). The mechanical nociceptive threshold (MNT) and cutaneous temperature (CT) values were collected at the same moments. The results show no intra- or intergroup differences, when evaluating each FE separately or in the GFS. In the intragroup comparison and 48 h after implantation, the control group showed higher values for SUM, PCoA, and SUM.W, although the horses implanted with polymers displayed more obvious alterations in the CT and MNT. Our findings show that the five statistical strategies used to analyze the faces of the horses were not able to detect low-grade inflammatory pain.
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