New insights into swine dysentery: faecal shedding, macro and microscopic lesions and biomarkers in early and acute stages of Brachyspira hyodysenteriae infection.

BACKGROUND: Swine dysentery (SD) is a severe mucohaemorrhagic colitis in pigs caused classically by Brachyspira hyodysenteriae. Although several aspects of B. hyodysenteriae infection dynamic are already described, further research in the early stage of this infection is required. In this study, 7-week-old pigs were orally challenged with B. hyodysenteriae to obtain information about faecal shedding, macro and microscopic intestinal lesions and serum acute phase proteins in pigs at the onset of B. hyodysenteriae shedding (early infection group, n = 8), in pigs with mucohaemorrhagic diarrhoea (acute infection group, n = 8) and in non-infected controls (n = 16). RESULTS: First B. hyodysenteriae detection by q-PCR and first loose stools with blood and mucus occurred both at 8 days post-inoculation. The lapse between a positive q-PCR and observation of mucohaemorrhagic diarrhoea ranged from 0 to 3 days, except in a single pig in which this period lasted 5 days. Macroscopic lesions were observed in the large intestine from both infected groups although more frequent and severe in acute infection group. Microscopic observation of the apex mucosa revealed that in early infection only higher ulceration values were observed compared to healthy controls. In contrast, the acute infection group exhibited higher ulceration, neutrophils infiltration and increased mucosal thickness compared to the other two groups. Among the serum biomarkers tested, only haptoglobin, C-reactive protein, and creatine kinase showed a significant increase in pigs in the acute infection period compared to controls, whereas haptoglobin was the only factor with a significant increase at the early infection compared to non-infected animals. CONCLUSIONS: This study provides new insights about SD and remarks the complex and limited options to perform an early detection of infected animals beyond PCR diagnosis.

Chromatin condensation but not DNA integrity of pig sperm is greater in the sperm-rich fraction.

BACKGROUND: Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo development. In some mammals, like pigs, ejaculates are emitted in three separate fractions: pre-sperm, sperm-rich (SRF) and post sperm-rich (PSRF). These fractions are known to vary in volume, sperm concentration and quality, as well as in the origin and composition of seminal plasma (SP), with differences being also observed within the SRF one. Yet, whether disparities in the DNA integrity and chromatin condensation and protamination of their sperm exist has not been interrogated. RESULTS: This study determined chromatin protamination (Chromomycin A3 test, CMA3), condensation (Dibromobimane test, DBB), and DNA integrity (Comet assay) in the pig sperm contained in the first 10 mL of the SRF (SRF-P1), the remaining portion of the sperm-rich fraction (SRF-P2), and the post sperm-rich fraction (PSRF). While chromatin protamination was found to be similar between the different ejaculate fractions (P > 0.05), chromatin condensation was seen to be greater in SRF-P1 and SRF-P2 than in the PSRF (P = 0.018 and P = 0.004, respectively). Regarding DNA integrity, no differences between fractions were observed (P > 0.05). As the SRF-P1 has the highest sperm concentration and ejaculate fractions are known to differ in antioxidant composition, the oxidative stress index (OSi) in SP, calculated as total oxidant activity divided by total antioxidant capacity, was tested and confirmed to be higher in the SRF-P1 than in SRF-P2 and PSRF (0.42 ± 0.06 vs. 0.23 ± 0.09 and 0.08 ± 0.00, respectively; P < 0.01); this index, in addition, was observed to be correlated to the sperm concentration of each fraction (Rs = 0.973; P < 0.001). CONCLUSION: While sperm DNA integrity was not found to differ between ejaculate fractions, SRF-P1 and SRF-P2 were observed to exhibit greater chromatin condensation than the PSRF. This could be related to the OSi of each fraction.

Preliminary Evaluation Salivary Biomarkers in Patients with Oral Potentially Malignant Disorders (OPMD): A Case-Control Study.

INTRODUCTION: Oral potentially malignant disorders (OPMD) are lesions associated with an increased risk of transformation (MT) into cancer. OBJECTIVE: A study was made of the salivary levels of adenosine deaminase (ADA), ferritin (FRR) and total proteins (TP) in healthy individuals and in patients with oral potentially malignant disorders (OPMD), assessing the potential role of saliva as a diagnostic tool. METHODS: A total of 91 subjects participated in the study, divided into two groups-59 patients with OPMD (oral leukoplakia or oral lichen planus) and 32 healthy controls-with measurements being made of salivary ADA, ferritin (FRR) and total proteins (TP). RESULTS: There were no significant differences in salivary mean ADA between the OPMD group 0.85 ± 2.18 UI/I and the controls 0.71 ± 1.72 UI/I (p = 0.934), though the levels of both FRR mean OPMD, 12.66 ± 10.50 (µg/L), versus control, 7.19 ± 4.44 (p = 0.001), and TP, 23.41 ± 17, versus control, 14.15 ± 15.19, were significantly higher in the OPMD group (p = 0.001). Patients with oral lichen planus showed significant differences in terms of FRR (p = 0.009) and TP (p = 0.003). The ferritin in LPO with a cut-off point of 8.5C showed a sensitivity and specificity of 54.3% and 82.3, respectively. The area under the curve (AUC) was 0.69 (95% confidence interval (95% CI): 0.58-0.82; p = 0.003). CONCLUSIONS: Ferritin and total proteins may constitute potential salivary biomarkers for oral lichen planus, though further studies are still needed in this field. In addition, saliva testing is a reliable and noninvasive diagnostic tool and appears to be a reliable strategy offering an interesting alternative for the screening of large populations.

Effect of the Probiotic Bacillus subtilis DE-CA9TM on Fecal Scores, Serum Oxidative Stress Markers and Fecal and Serum Metabolome in Healthy Dogs.

BACKGROUND: There is increasing interest in the use of Bacillus species as probiotics since their spore-forming ability favors their survival in the acidic gastric environment over other probiotic species. The subsequent germination of B. subtilis to their vegetative form allows for their growth in the small intestine and may increase their beneficial effect on the host. B. subtilis strains have also previously been shown to have beneficial effects in humans and production animals, however, no reports are available so far on their use in companion animals. STUDY DESIGN: The goal of this study was therefore to investigate the daily administration of 1 × 109 cfu DE-CA9TM orally per day versus placebo on health parameters, fecal scores, fecal microbiome, fecal metabolomics, as well as serum metabolomics and oxidative stress markers in ten healthy Beagle dogs in a parallel, randomized, prospective, placebo-controlled design over a period of 45 days. RESULTS: DE-CA9TM decreased the oxidative status compared to controls for advanced oxidation protein products (AOPP), thiobarbituric acid reactive substances (TBARS) and reactive oxygen metabolites (d-ROMS), suggesting an antioxidant effect of the treatment. Fecal metabolomics revealed a significant reduction in metabolites associated with tryptophan metabolism in the DE-CA9TM-treated group. DE-CA9TM also significantly decreased phenylalanine and homocysteine and increased homoserine and threonine levels. Amino acid metabolism was also affected in the serum metabolome, with increased levels of urea and cadaverine, and reductions in N-acetylornithine in DE-CA9TM compared to controls. Similarly, changes in essential amino acids were observed, with a significant increase in tryptophan and lysine levels and a decrease in homocysteine. An increase in serum guanine and deoxyuridine was also detected, with a decrease in beta-alanine in the animals that ingested DE-CA9TM. CONCLUSIONS: Data generated throughout this study suggest that the daily administration of 1 × 109 cfu of DE-CA9TM in healthy Beagle dogs is safe and does not affect markers of general health and fecal scores. Furthermore, DE-CA9TM administration had a potential positive effect on some serum markers of oxidative stress, and protein and lipid metabolism in serum and feces.

The Impact of Epigallocatechin Gallate and Coconut Oil Treatment on Cortisol Activity and Depression in Multiple Sclerosis Patients.

(1) Background: Multiple sclerosis (MS) is pathogenically characterized by high oxidative stress and symptomatically by progressive muscle loss and increased body fat associated with the presence of depression. Epigallocatechin gallate (EGCG) (particularly present in green tea) and ketone bodies (in particular beta-hydroxybutyrate (BHB)), whose main source is coconut oil, have shown emotional benefits and body fat loss. The aim of this study was to assess the impact of EGCG and coconut oil on cortisol activity related to fat loss and depression in MS patients. (2) Methods: The study involved 51 MS patients who were randomly divided into an intervention group or a control group. The intervention group received 800 mg of EGCG and 60 mL of coconut oil, which were included in their daily diet for four months. The control group received placebo and all patients followed an isocaloric diet. A blood sample was collected before and after the four-month period, and levels of cortisol, albumin and BHB were measured in serum. In addition, immediately before and after the intervention, anthropometric variables were measured: waist-to-hip ratio (WHR), body fat mass percentage, fat weight, total weight, and muscle mass percentage. Depression was assessed with the Beck Depression Inventory II (BDI-II). (3) Results: No significant changes were obtained in cortisol levels in any of the groups, and there was a significant increase in albumin in the blood of the intervention group only that could lead to a decrease in serum free cortisol. In addition, it was observed a significant decrease in levels of depression and abdominal fat. (4) Conclusions: EGCG combined with coconut oil increase the concentration of albumin in blood and produce less depression in MS patients.

Effects of Commercial Antioxidants in Feed on Growth Performance and Oxidative Stress Status of Weaned Piglets.

This work aimed to evaluate the effect of adding two different commercial antioxidants (AOX) products to pre-starter and starter diets using low vitamin E (Vit E as DL-α-tocopheryl acetate) levels on the growth performance and oxidative stress of piglets for the first six weeks post-weaning (PW). They were sorted by initial body weight (BW: 6.175 ± 0.931 kg) and randomly allotted to four dietary treatments (with six replicates per treatment): a positive control (PC) and a negative control (NC) diet, with normal and low dose of vitamin E (80 and 15 mg kg-1, respectively), both without AOX; the other two experimental diets with a low dose of vitamin E (LVE) plus LOXIDAN VD100 (LVE + AOX1) or LOXIDAN E Ros (LVE + AOX2). Growth data were recorded, and blood samples were taken, at the beginning (day 0) and at the end of each feeding period: pre-starter and starter (at days 14 and 42, respectively). No differences among dietary treatments were found with respect to growth performance in the pre-starter period (p ≥ 0.05). However, at the end of the starter period, a lower BW was found in piglets fed the NC diet compared to the other dietary treatments. Differences in daily gain and feed conversion ratio were also found either for the starter period or when the whole period was considered (p < 0.05), whereby piglets fed PC or LVE diets supplemented with AOX showed better growth performance compared to piglets fed the NC diet. Regarding Vit E (α-tocopherol) serum levels, there were no differences among treatments at day 0; but the serum values of this vitamin decreased in LVE diets at 14 and 42 days, but not in the PC. On day 42, the highest levels of α-tocopherol in liver were also found in piglets fed PC (p < 0.05). Nevertheless, in general, from a metabolic point of view and after checking the serum biochemical profile of piglets, there were no differences in other oxidative stress markers (p ≥ 0.05). The results showed that the AOX products used were able to compensate for the lower Vit E supply with respect to growth performance in the starter phase. The use of AOXs or usual levels of Vit E in feed constitutes a key factor in achieving optimal growth performance of piglets in the PW period.

Changes in Serum Biomarkers of Oxidative Stress in Cattle Vaccinated with Tick Recombinant Antigens: A Pilot Study.

Tick vaccination is an environmentally friendly alternative for tick control, pathogen infection, and transmission. Tick vaccine protection is sometimes incomplete, which may be due to problems in the stability, conformation, and activity of antibodies. This might be related to oxidative stress, but more studies are needed about the possible relationships between oxidative stress and immune function. The objective of this study was to evaluate and compare various serum biomarkers of antioxidant response and oxidative damage in cattle vaccinated with two recombinant antigens, the chimera of Subolesin- BM95 (homologue antigen of BM86)-MSP1a and BM86, and a control consisting in the adjuvant of the vaccines. Cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of the plasma (FRAP), trolox equivalent antioxidant capacity (TEAC), total thiol concentrations, and uric acid were evaluated in serum to determine the antioxidant response. To evaluate oxidative status, ferrous oxidation-xylenol orange (FOX), total oxidant status (TOS), advanced oxidation protein products (AOPP) and hydrogen peroxide (H2O2) concentrations in serum were determined. In addition, correlations between biomarkers of oxidative stress and antibody titers were evaluated. A significant decrease in all antioxidant biomarkers, with exception of thiol, and also a decrease in the oxidant markers TOS, AOPP and H2O2 was observed in cattle vaccinated with BM86, that also showed the highest antibody titers response whereas no significant differences in any of the biomarkers were detected in the Subolesin-Bm95-MSP1a and control groups. In addition, the dynamics of Cuprac and H2O2 with time showed significant differences between the groups. Although this is a pilot study and the results should be interpreted with caution and corroborated by studies involving a large number of animals, our results indicate that, in our experimental conditions, those vaccines able to induce a lower oxidative stress produce a higher concentration of antigen-specific antibodies. Overall, the results of the study provided information on the behavior of different biomarkers related to antioxidant defense, and the oxidative damage in cattle in response to vaccination.

Effects of Dietary Supplementation of Garlic and Oregano Essential Oil on Biomarkers of Oxidative Status, Stress and Inflammation in Postweaning Piglets.

The effects of two different concentrations of micro capsuled oregano essential oil (OEO) and purple garlic powder on biomarkers of oxidative status, stress, and inflammation, as well as on average daily gain (ADG) and feed conversion ratio (FCR), were evaluated in piglets during the postweaning period. The trial was carried out with 300 crossbred pigs of 21 days of age fed with different concentrations of OEO and purple garlic powder and ZnO. Saliva and serum samples were taken to evaluate a panel of biomarkers of oxidative status, stress, and inflammation. OEO and garlic powder at 0.4% did not produce significant changes in C-reactive protein (CRP) and cortisol and yielded higher levels of the antioxidant biomarker CUPRAC in serum than higher doses (p < 0.01); they yielded a better ADG than the control and ZnO diets. OEO and garlic powder at higher concentrations than 0.4% showed higher concentrations of CRP (p < 0.05). Overall, doses of OEO and garlic powder at 0.4% did not lead to inflammation, stress, or negative changes in oxidative biomarkers in piglets during the postweaning period and gave better productive performance than the control and ZnO diets. High doses of OEO and garlic powder were ineffective and could negatively affect the animals. Therefore, our results highlight the importance of the dose used when OEO or garlic are supplemented to piglets.