An Approach for Engineering Peptides for Competitive Inhibition of the SARS-COV-2 Spike Protein.

SARS-CoV-2 is the virus responsible for a respiratory disease called COVID-19 that devastated global public health. Since 2020, there has been an intense effort by the scientific community to develop safe and effective prophylactic and therapeutic agents against this disease. In this context, peptides have emerged as an alternative for inhibiting the causative agent. However, designing peptides that bind efficiently is still an open challenge. Here, we show an algorithm for peptide engineering. Our strategy consists of starting with a peptide whose structure is similar to the interaction region of the human ACE2 protein with the SPIKE protein, which is important for SARS-COV-2 infection. Our methodology is based on a genetic algorithm performing systematic steps of random mutation, protein-peptide docking (using the PyRosetta library) and selecting the best-optimized peptides based on the contacts made at the peptide-protein interface. We performed three case studies to evaluate the tool parameters and compared our results with proposals presented in the literature. Additionally, we performed molecular dynamics (MD) simulations (three systems, 200 ns each) to probe whether our suggested peptides could interact with the spike protein. Our results suggest that our methodology could be a good strategy for designing peptides.

Factors That Most Influence the Choice for Fast Food in a Sample of Higher Education Students in Portugal.

The frequency of fast food consumption among higher education students is high, causing worrying implications for public health. This study aims to relate the factors that influence the choice for fast food with social factors, nutritional status, and fast food consumption in a sample of higher education students in Portugal. An online questionnaire was developed and disseminated by social networks among students during the first half of 2023. Two hundred and thirty-seven students participated, mainly female (65.4%), who were attending public higher education institutions (59.1%), with a median of age of 20.0 (19.0; 22.0) years, and about 20% of the sample had overweight. Approximately 80% consumed fast food, and almost 40% consumed it more than once or twice a week. Predominantly (78.0%), they chose hamburger meals, spending EUR 8.0 per meal. The factors that most influenced the choice of fast food were ease or convenience of preparation (59.9%), price (48.5%), and flavor (28.3%). There were also differences between sexes and between those attending public and private higher education institutions regarding whether they usually consume fast food or not. The body mass index was positively associated with age (r: 0.142; p = 0.029) and with fast food spending (r: 0.146; p = 0.024). The results have implications for public health and clinical nutrition, and can support more effective strategies to improve food choices in higher education students.

Assessment of color changes and adverse effects of over-the-counter bleaching protocols: a systematic review and network meta-analysis.

OBJECTIVES: To assess color change efficacy and the adverse effects of varied over-the-counter (OTC) bleaching protocols. METHODOLOGY: The study included randomized clinical trials evaluating color changes from OTC bleaching agents. Nine databases were searched, including the partial capture of the grey literature. The RoB2 tool analyzed the individual risk of bias in the studies. Frequentist network meta-analyses compared treatments through common comparators (∆Eab* and ∆SGU color changes, and tooth sensitivity), integrating direct and indirect estimates and using the mean and risk differences as effect measures with respective 95% confidence intervals. The GRADE approach assessed the certainty of the evidence. RESULTS: Overall, 37 remaining studies constituted the qualitative analysis, and ten composed the meta-analyses. The total sample included 1,932 individuals. ∆Eab* was significantly higher in groups 6% hydrogen peroxide (HP) strips (≥ 14 h). ∆SGU was significantly higher in groups at-home 10% carbamide peroxide (CP) (≥ 14 h), followed by 6% HP strips (≥ 14 h) and 3% HP strips (≥ 14 h). At-home 10% CP (7-13 h) and placebo showed lower risks of tooth sensitivity without significant differences between these treatments. CONCLUSION: Considering the low level of evidence, OTC products presented satisfactory short-term effects on tooth bleaching compared to the placebo, with little to no impact on dentin hypersensitivity and gingival irritation. CLINICAL RELEVANCE: OTC products are proving to be practical alternatives for tooth whitening. However, patients should be advised about the possible risks of carrying out such procedures without professional supervision.

Annexin A1 improves immune responses and control of tissue parasitism during Leishmania amazonensis infection in BALB/c mice.

Leishmaniases, a group of diseases caused by the species of the protozoan parasite Leishmania, remains a significant public health concern worldwide. Host immune responses play a crucial role in the outcome of Leishmania infections, and several mediators that regulate inflammatory responses are potential targets for therapeutic approaches. Annexin A1 (AnxA1), an endogenous protein endowed with anti-inflammatory and pro-resolving properties, has emerged as a potential player. We have shown that during L. braziliensis infection, deficiency of AnxA1 exacerbates inflammatory responses but does not affect parasite burden. Here, we have investigated the role of AnxA1 in L. amazonensis infection, given the non-healing and progressive lesions characteristic of this infectious model. Infection of AnxA1 KO BALB/c mice resulted in increased lesion size and tissue damage associated with higher parasite burdens and enhanced inflammatory response. Notably, therapeutic application of the AnxA1 peptidomimetic Ac2-26 improves control of parasite replication and increases IL-10 production in vivo and in vitro, in both WT and AnxA1 KO mice. Conversely, administration of WRW4, an inhibitor of FPR2/3, resulted in larger lesions and decreased production of IL-10, suggesting that the effects of AnxA1 during L. amazonensis infection are associated with the engagement of these receptors. Our study illuminates the role of AnxA1 in L. amazonensis infection, demonstrating its impact on the susceptibility phenotype of BALB/c mice. Furthermore, our results indicate that targeting the AnxA1 pathway by using the Ac2-26 peptide could represent a promising alternative for new treatments for leishmaniasis.

Does Faeces Excreted by Moxidectin-Treated Sheep Impact Coprophagous Insects and the Activity of Soil Microbiota in Subtropical Pastures?

Moxidectin (MOX) is used to control helminth parasites in ruminant livestock. It is released through feces and remains in the environment for a long period. This study aimed to evaluate the impact of faeces excreted by moxidectin-treated sheep on soil biodiversity (coprophagous insects, soil microbial biomass, and activity) to establish environment-related guidelines regarding the use of MOX in sheep livestock. The study consisted of two experiments. In the first one, faeces from MOX-treated (subcutaneous dose of 0.2 mg·kg-1 body weight) and nontreated rams were placed on an animal-free pasture field, protected or not against rain, for 88 days. Then, coprophagous insects were captured, identified, and counted, and faeces degradation was evaluated by measuring dry weight and carbon (C) and nitrogen (N) contents over time. Diptera, Hymenoptera, Isoptera, and Coleoptera were equally encountered in faeces from MOX-treated and nontreated animals. Faecal boluses of MOX-treated animals (with higher N content) not protected against rain degraded faster than faecal boluses of nontreated animals (with lower N content). In the second experiment, faeces from nontreated animals were amended with increasing amounts of MOX (75 to 3,000 ng·kg-1 faeces), mixed with soil samples from animal-free pasture (1.9 to 75 ng·kg-1 soil), and incubated in a greenhouse for 28 days. Increasing concentrations of MOX did not prevent the growth of cultivable bacteria, actinobacteria, or fungi in culture media. However, even the lower MOX concentration (1.9 ng·kg-1 soil) abruptly decreased soil microbial biomass, basal respiration, and N mineralization. Thus, the results indicate that faeces excreted from sheep treated with MOX under the experimental conditions of this study are not harmful to the coprophagous insects. However, adding MOX to faeces from drug-free sheep had a negative impact on soil microbial activity and biomass.

The efficacy of combining adjuvants with non-surgical periodontal therapy in individuals with type 2 diabetes: A Bayesian network meta-analysis.

AIM: This Bayesian network meta-analysis of randomized controlled trials assessed the effect of adjuvant periodontal treatment in both periodontal and HbA1c outcomes in adult individuals with type 2 diabetes (T2DM). MATERIALS AND METHODS: A systematic search was done up to February 2023 comparing sub-gingival debridement (SD) in combination with local or systemic adjuvant treatment with SD alone for individuals with T2DM. The primary outcomes were changes in absolute HbA1c levels and full-mouth probing depth reported at 3- to 6-month post-treatment. RESULTS: Seventy-two eligible publications evaluating 27 adjuvant treatments were retrieved. The combination of SD and systemic antibiotic metronidazole or SD and antioxidant alpha lipoic acid provided, respectively, 1.4% (95% credible interval [CrI] 0.48; 2.20) and 2.4% (95% CrI 1.50; 3.30) more significant improvement on HbA1c levels, and 0.89 mm (95% CrI 0.23; 1.50) and 0.92 mm (95% CrI 0.02; 0.92) greater periodontal probing depth reductions. Other adjuvant treatments provided added benefit to the periodontal outcomes without discernible effects on HbA1c. CONCLUSIONS: Adjuvant use of metronidazole or alpha lipoic acid was the best adjunct option to provide clinically meaningful HbA1c levels and probing depth reductions. However, no strong recommendation can be drawn due to the scarcity of studies for each adjuvant treatment and the low certainty of the resultant evidence.

Development of a new assay for quantification of parasite load of intracellular Leishmania sp. in macrophages using flow cytometry.

Finding novel methodologies that enhance the precision, agility, and standardization of drug discovery is crucial for studying leishmaniasis. The slide count is the technique most used to assess the leishmanicidal effect of a given drug in vitro. Despite being consolidated in the scientific environment, it presents several difficulties in its execution, assessment, and results. In addition to being laborious, this technique takes time, both for the preparation of the material for analysis and for the counting itself. Our research group suggests a fresh approach to address this requirement, which involves utilizing nuclear labeling with propidium iodide and flow cytometry to determine the quantity of Leishmania sp. parasites present in macrophages in vitro. Our results show that the fluorescence of infected samples increases as the infection rate increases. Using Pearson's Correlation analysis, it was possible to establish a correlation coefficient (Pearson r = 0.9473) that was strongly positive, linear, and directly proportional to the fluorescence and infection rate variables. Thus, it is possible to infer a mathematical equation through linear regression to estimate the number of parasites in each sample using the Relative Fluorescence Units (RFU) values. This new methodology opens space for the possibility of using this methodological resource in the in vitro quantification of Leishmania in macrophages.

Diagnostic accuracy of ultrasonography in relation to salivary gland biopsy in Sjögren's syndrome: a systematic review with meta-analysis.

OBJECTIVES: To evaluate the accuracy of major salivary gland ultrasonography (SGUS) in relation to minor salivary gland biopsy (mSGB) in the diagnosis of Sjögren's syndrome (SS). METHODS: A systematic review and meta-analysis were performed. Ten databases were searched to identify studies that compared the accuracy of SGUS and mSGB. The risk of bias was assessed, data were extracted, and univariate and bivariate random-effects meta-analyses were done. RESULTS: A total of 5000 records were identified; 13 studies were included in the qualitative synthesis and 10 in the quantitative synthesis. The first meta-analysis found a sensitivity of 0.86 (95% CI: 0.74-0.92) and specificity of 0.87 (95% CI: 0.81-0.92) for the predictive value of SGUS scoring in relation to the result of mSGB. In the second meta-analysis, mSGB showed higher sensitivity and specificity than SGUS. Sensitivity was 0.80 (95% CI: 0.74-0.85) for mSGB and 0.71 (95% CI: 0.58-0.81) for SGUS, and specificity was 0.94 (95% CI: 0.87-0.97) for mSGB and 0.89 (95% CI: 0.82-0.94) for SGUS. CONCLUSIONS: The diagnostic accuracy of SGUS was similar to that of mSGB. SGUS is an effective diagnostic test that shows good sensitivity and high specificity, in addition to being a good tool for prognosis and for avoiding unnecessary biopsies. More studies using similar methodologies are needed to assess the accuracy of SGUS in predicting the result of mSGB. Our results will contribute to decision-making for the implementation of SGUS as a diagnostic tool for SS, considering the advantages of this method.

Adverse alcohol use and oral health.

OBJECTIVE: To determine whether different alcohol intake dimensions: average alcohol volume consumed (AVC), binge drinking (BD), and alcohol-related consequences (ARC) are associated with self-rated oral health (SROH). METHODS: Secondary cross-sectional analysis of The Brazilian National Health Survey of 2019 data. BD was considered when an individual reported a past-month heavy drinking episode. ARC referred to experiences such as past-year blackouts, concerns from others, or failure in routine activities. Adjusted multivariate Poisson regression models with robust variance were employed to calculate the prevalence ratios (PR) and 95% confidence intervals (CI) of SROH across the entire sample and stratified by sex and age. RESULTS: The sample comprised 88,531 participants aged 18 years or older. Individuals experiencing at least one past-year ARC had an 11% (95% CI: 1.07, 1.15) higher prevalence of poor SROH than their abstainers' counterparts. Associations were more pronounced among men aged less than 50 (PR = 1.09; 95% CI: 1.03, 1.15) and women aged 50 years or more (PR = 1.15; 95% CI: 1.01, 1.30). CONCLUSION: Alcohol-related consequences predicted poor SROH. Oral health inquiries should include the screening for ARC in addition to traditional measurements of AVC or BD, since such exposures may not fully capture the role of alcohol on oral health impairment.

Bionanocomposite Based on Cassava Waste Starch, Locust Bean Galactomannan, and Cassava Waste Cellulose Nanofibers.

Natural polysaccharides are among the renewable sources with great potential for replacing petroleum-derived chemicals as precursors to produce biodegradable films. This study aimed to prepare biopolymeric films using starch extracted from the periderm and cortex of cassava roots (waste from cassava root processing), locust bean galactomannan, and cellulose nanofibers also obtained from cassava waste. The films were prepared by casting, and their physicochemical, mechanical, and biodegradability properties were evaluated. The content of cellulose nanofibers varied from 0.5 to 2.5%. Although the addition of cellulose nanofibers did not alter the mechanical properties of the films, it significantly enhanced the vapor barrier of the films (0.055 g mm/m2 h kPa-2.5% nanofibers) and their respective stabilities in aqueous acidic and alkaline media. All prepared films were biodegradable, with complete degradation occurring within five days. The prepared films were deemed promising alternatives for minimizing environmental impacts caused by the disposal of petroleum-derived materials.

The effect of body mass index on maternal and perinatal outcomes in COVID-19 infection during pregnancy and postpartum: Secondary analysis from the REBRACO cohort study.

OBJECTIVES: To compare maternal and perinatal outcomes among women with obesity, overweight, and normal body mass index, associated with COVID-19 infection during pregnancy and postpartum. METHOD: Prospective Cohort Study, within the REBRACO (Brazilian Network of COVID-19 in Pregnancy) multicenter initiative. Confirmed positive cases of SARS-CoV-2 were included, and women categorized into three groups according to their pre-pregnancy BMI: obesity (BMI ≥ 30), overweight (BMI <30 but >25), and normal BMI. Sociodemographic, clinical, and obstetric characteristics and different maternal and perinatal outcomes were compared, and a multiple regression analysis was performed to investigate factors independently associated with adverse maternal and perinatal outcomes. RESULTS: Two hundred eighty-nine women positive for SARS-CoV-2 infection were considered, and 202 had available data on maternal BMI for the current analysis. Overall, 72 (35.6%)obese, 68 (33.6%) overweight, and 60 (29.7%) normal BMI. Obesity was associated with increased adverse clinical outcomes including sepsis (P = 0.02), acute respiratory distress syndrome (P = 0.002), and the need for mechanical ventilation (P = 0.044). Considering perinatal outcomes, a multiple regression model confirmed obesity as an independent factor associated with adverse results (adjusted odds ratio 3.73, 95% CI 1.54-9.08). CONCLUSION: Obesity and overweight were associated with worse clinical outcomes, severe/critical COVID-19, and adverse perinatal outcomes.

Should supragingival scaling be performed separately prior to subgingival scaling and root planning in nonsurgical periodontal therapy? A systematic review of randomized trials.

OBJECTIVE: To systematically evaluate randomized controlled trials (RCTs), with at least 6 months of follow-up, on whether professional mechanical plaque removal (PMPR) including supragingival scaling should be performed prior and separately from subgingival scaling and root planning (SRP) in nonsurgical periodontal therapy (NSPT), in terms of clinical and patient-reported outcomes (PROs) (CRD42020219759). METHODS: The MEDLINE, EMBASE, CENTRAL, LILACS and Web of Science electronic databases, as well as grey literature sources, were searched by two independent reviewers up to May 2023. The Cochrane Collaboration's Risk of Bias tool (RoB 2.0) was used for quality appraisal and GRADE for assessing the certainty of evidence. Random-effects pairwise meta-analyses compared the changes in probing pocket depth (PPD), clinical attachment loss (CAL), and bleeding on probing (BoP) of a stepwise NSPT approach (PMPR prior and separately from SRP) and conventional one-step NSPT through mean differences (MDs) and associated confidence intervals (95% CI). RESULTS: Two RCTs were included, including data of 77 participants with severe periodontitis. One RCT presents high risk of bias and the other has some concerns. No significant differences were found between the stepwise approach and performing both steps simultaneously for any clinical outcomes, with overall very low certainty on evidence. No adverse effects were detected and there was no data on PROs. CONCLUSIONS: There is very-low certainty evidence of no significant difference on PPD and BoP reductions and CAL gain between supragingival scaling performed prior and separately from SRP and conventional one-step NSPT.

Impact of Early Arsenic Exposure on the Mineral Content and Oxidative Status of the Liver and Kidney of Pubescent and Adult Rats.

This study evaluated the effect of prepubertal arsenic exposure in the liver and kidney of pubescent rats and their reversibility 30 days after arsenic withdrawal. Male pups of Wistar rats (21 days old) were divided into two groups (n = 20/group): control animals received filtered water, and exposed rats received 10 mg L-1 arsenic from postnatal day (PND) 21 to PND 51. The liver and kidney of 52 days old rats (n = 10/group) were examined to investigate the effects of arsenic on micromineral content, antioxidant enzyme activity, histology, and biochemistry parameters. The other animals were kept alive under free arsenic conditions until 82 days old and further analyzed by the same parameters. Our results revealed that 52-day-old rats increased arsenic content in their liver and arsenic and manganese in their kidney. In those animals, glycogen and zinc content and catalase activity were reduced in the liver, and the selenium content decreased in the kidney. Thirty days later, arsenic reduced the manganese and iron content and SOD and CAT activity in the liver of 82-day-old rats previously exposed to arsenic, while glycogen and selenium content decreased in their kidney. In contrast, PND 82 rats exhibited higher retention of copper in the liver, an increase in iron and copper content, and CAT and GST activity in the kidney. Significant histological alterations of liver and kidney tissues were not observed in rats of both ages. We conclude that arsenic-induced toxicity could alter differently the oxidative status and balance of trace elements in pubertal and adult rats, demonstrating that the metalloid can cause effects in adulthood.

Immunisation with Neospora caninum subunits rsNcSAG4 and rsNcGRA1 (NcSAG4 and NcGRA1 epitopes construct) in BALB/c mice: the profile of the immune response and controlling the vertical transmission.

Neospora caninum is an apicomplexan protozoan that causes neosporosis, which has a high economic impact on cattle herds with no available vaccine. During infection, the secretion of dense granules and the expression of surface antigens play an important role in hosting immunomodulation. However, some epitopes of those antigens are immunogenic, and using these fractions could improve the subunit antigens in vaccine design. This study evaluates the recombinant peptides rsNcGRA1 and rsNcSAG4 derived from NcGRA1 and NcSAG4 native antigens as vaccine candidates produced by a fermentative process in the yeast culture system of Komagataella phaffii strain Km71, confirmed by colony PCR, SDS-PAGE, and western blotting. The assay was conducted in BALB/c mice using the peptides at low (25 µg) and standard (50 µg) dosages in monovalent and combined administrations at three time points with saponin as an adjuvant assessing the immunogenicity by antibodies response and cytokine production. We challenge the females after pregnancy confirmation using 2 × 105 NC-1 tachyzoites previously propagated in Vero cells. We assessed the chronic infection in dams and vertical transmission in the offspring by PCR and histopathology. Mice, especially those immunised with combined peptides and monovalent rsNcGRA1 at a standard dose, controlling the chronic infection in dams with the absence of clinical manifestations, showed an immune response with induction of IgG1, a proper balance between Th1/Th2 cytokines and reduced vertical transmission in the pups. In contrast, dams inoculated with a placebo vaccine showed clinical signs, low-scored brain lesions, augmented chronic infection with 80% positivity, 31% mortality in pups, and 81% vertical transmission. These findings indicate that rsNcGRA1 peptides in monovalent and combined with rsNCSAG4 at standard dose are potential vaccine candidates and improve the protective immune response against neosporosis in mice.

The role of gut-brain axis in a rotenone-induced rat model of Parkinson's disease.

Parkinson's disease (PD) is a widespread neurodegenerative condition affecting millions globally. This investigation centered on the gut-brain axis in a rotenone-induced PD rat model. Researchers monitored behavioral shifts, histological modifications, neurodegeneration, and inflammation markers throughout the rats' bodies. Results revealed that rotenone-treated rats displayed reduced exploration (p = 0.004) and motor coordination (p < 0.001), accompanied by decreased Nissl staining and increased alpha-synuclein immunoreactivity in the striatum (p = 0.009). Additionally, these rats exhibited weight loss (T3, mean = 291.9 ± 23.67; T19, mean = 317.5 ± 17.53; p < 0.05) and substantial intestinal histological alterations, such as shortened villi, crypt architecture loss, and inflammation. In various regions, researchers noted elevated immunoreactivity to ionized binding adapter molecule (IBA)-1 (p < 0.05) and reduced immunoreactivity to glial fibrillary acidic protein (p < 0.05) and S100B (p < 0.001), indicating altered glial cell activity. Overall, these findings imply that PD is influenced by gut-brain axis changes and may originate in the intestine, impacting bidirectional gut-brain communication.

Do blood concentrates influence inflammatory signs and symptoms after mandibular third molar surgery? A systematic review and network meta-analysis of randomized clinical trials.

OBJECTIVES: To investigate, through a network meta-analysis, the effectiveness of blood concentrates in reducing pain perception, trismus, and edema after mandibular third molar extraction. MATERIALS AND METHODS: An electronic search was performed in nine databases to locate randomized clinical trials comparing blood concentrate use after mandibular third molar extraction. Two authors selected and extracted the data independently. The individual risk of bias in the studies was assessed with the RoB v2.0 tool. A network meta-analysis compared postoperative pain and trismus scores after applying different blood concentrates, using the mean difference (MD) as an effect estimate. The GRADE approach assessed the certainty of evidence. RESULTS: Thirty-one randomized clinical trials were included in the review and 18 in the meta-analysis. Leukocyte- and platelet-rich fibrin (L-PRF) was the most used blood concentrate, followed by platelet-rich plasma (PRP). The network meta-analysis, depending on the analyzed period, evaluated up to 1240 surgeries. Among the analyzed blood concentrates, advanced platelet-rich fibrin (A-PRF) performed better among the analyzed blood concentrates, decreasing postoperative pain in 1, 2, 3, and 7 days and reducing trismus up to the inflammatory peak compared to blood clots. Only two studies had a low risk of bias. CONCLUSIONS: Based on very low certainty of evidence, using concentrates seemed efficient compared to blood clots in reducing pain and trismus after mandibular third molar surgeries. A-PRF decreased postoperative pain throughout the evaluated time and trismus during the acute inflammatory peak. CLINICAL RELEVANCE: A-PRF after mandibular third molar extractions performed better among the analyzed blood concentrates and seemed efficient in improving postoperative quality by decreasing inflammatory signs and symptoms.

Consumption of yacon flour and energy-restricted diet increased the relative abundance of intestinal bacteria in obese adults.

Prebiotics can alter the gastrointestinal environment, favoring the growth of health-promoting bacteria. Although yacon is a functional food, with prebiotic properties (fructooligosaccharides), its effects on the intestinal microbiota have not been investigated yet. The objective of this study was to evaluate the effects of yacon flour consumption and energy-restricted diet in the intestinal microbiota in adults with excess body weight. Twenty-one adults with excess body weight were included in this randomized, parallel, double-blind, placebo-controlled, 6-week clinical trial. Subjects daily consumed at breakfast a drink containing 25 g of yacon flour (n = 11) or not containing yacon (n = 10) and received the prescription of energy-restricted diets. Fecal samples were collected on the first and on last day of the study. 16S rRNA sequencing was assessed to evaluate the effect of yacon fermentation on intestinal microbiota bacterial composition. There was an increase in the genera Bifidobacterium, Blautia, Subdoligranulum, and Streptococcus after the consumption of yacon and energy-restricted diet. In the yacon group, we also observed a positive correlation between the concentrations of short-chain fatty acids versus the genera Coprococcus and Howardella, besides a negative correlation between the concentrations of advanced glycation end products and early glycation products versus the genera Ruminococcus and Prevotella, respectively. Consumption of yacon flour and energy-restricted diet selectively changed the intestinal microbiota composition in adults with excess body weight. TRIAL REGISTRATION: Register number: RBR-6YH6BQ. Registered 23 January, 2018.

Effect of antibiotics and low-crude protein diets on growth performance, health, immune response, and fecal microbiota of growing pigs.

This study aimed to investigate the effects of diets with and without antibiotics supplementation and diets with 18.5% and 13.0% crude protein (CP) on growth performance, carcass characteristics, disease incidence, fecal microbiota, immune response, and antioxidant capacity of growing pigs. One hundred and eighty pigs (59-day-old; 18.5 ±â€…2.5 kg) were distributed in a randomized complete block design in a 2 × 2 factorial arrangement, nine replicates, and five pigs per pen. The factors were CP (18.5% or 13.0%) and antibiotics (none or 100 mg/kg tiamulin + 506 mg/kg oxytetracycline). Medicated diets were fed from days 59 to 73. After that, all pigs were fed their respective CP diets from 73 to 87 days. Data were analyzed using the Mixed procedure in SAS version 9.4. From days 59 to 73, pigs fed antibiotics diets had higher (P < 0.05) average daily feed intake (ADFI), average daily weight gain (ADG), gain to feed ratio (G:F), compared to the diets without antibiotics. From days 73 to 87 (postmedicated period), any previous supplementation of antibiotics did not affect pig growth performance. Overall (days 59 to 87), pigs-fed antibiotics diets had higher (P < 0.05) G:F compared to pigs-fed diets without antibiotics. In all periods evaluated, pigs fed 18.5% CP diets had higher (P < 0.05) ADG and G:F compared to pigs fed 13.0% CP. Pigs fed the 13.0% CP diets had lower (P < 0.05) fecal score and diarrhea incidence than those fed 18.5% CP. Pigs fed 18.5% CP diets had improved (P < 0.05) loin area compared to pigs-fed diets with 13.0% CP. At 66 days of age, pigs-fed antibiotics diets had lower (P < 0.05) alpha diversity estimated with Shannon and Simpson compared to the pig-fed diets without antibiotics. At family level, pigs fed 18.5% CP diets had higher (P < 0.05) relative abundance of Streptococcaceae, and lower (P < 0.05) relative abundance of Clostridiaceae at days 66 and 87 compared with pigs fed 13.0% CP. Pigs-fed antibiotics diets had lower (P < 0.05) immunoglobulin G and protein carbonyl concentrations at day 66 compared to the pigs-fed diets without antibiotics. The reduction of dietary CP from 18.5% to 13.0% reduced the growth performance and loin muscle area of growing pigs, although it was effective to reduce diarrhea incidence. Antibiotics improved growth performance, lowered diarrhea incidence, improved components of the humoral immune response, and reduced microbiota diversity. However, in the postmedicated period, we found no residual effect on the general health of the animals, and considering the overall period, only G:F was improved by the use of antibiotics.

Dietary antibiotics have been used in pig farming practices to avoid health problems and improve animal growth performance. However, their use in production animals is considered a global health challenge, due to its association with selection of resistance in zoonotic bacteria. Another negative impact of pig farming that has gained attention is related to environmental pollution due to the excretion of nitrogenous compounds. Reducing dietary crude protein content has become a goal in the pig feed industry due to the limited availability and high cost of dietary protein sources, as well as the aim of enhancing gut health in pigs. Thus, the aim of this study was to investigate the effects of diets with and without antibiotics supplementation and diets with 18.5% and 13.0% crude protein for pigs. The reduction of dietary crude protein in this study reduced growth performance, although it was effective to reduce diarrhea incidence. Antibiotics improved growth performance, positively affected the overall health of animals, and reduced microbiota diversity. However, during the postmedicated period, we found no residual effect on the general health of the animals, and considering the overall period, only gain to feed ratio was improved by the use of antibiotics.