Agronomic, Nutritional Traits, and Alkaloids of Lupinus albus, Lupinus angustifolius and Lupinus luteus Genotypes: Effect of Sowing Dates and Locations.

Lupins (Lupinus spp.) are legumes with high relevance for the sustainability of agricultural systems as they improve the soil quality, namely, through the fixation of atmospheric nitrogen, and have good adaptability to different climates and soil conditions. Besides, they possess high nutritive value, especially due to the high protein content of the seeds. Nevertheless, the plants' productivity and metabolism can be influenced by the genotype, the edaphoclimatic conditions, and the sowing practices. In this work, the effect of edaphoclimatic conditions and sowing dates on the productivity, nutritional factors, and alkaloids of the seeds of L. albus cv. Estoril, L. angustifolius cv. Tango, and L. luteus cv. Cardiga was evaluated. High variability in the seeds and protein productions, nutritional traits, and alkaloid content related to the species was observed, along with a significant effect of the location. Lupinus albus cv. Estoril showed a good compromise between productivity and low alkaloid content, being an interesting genotype for food and feed use in the conditions of this trial.

Environmental conditions affect the nutritive value and alkaloid profiles of Lupinus forage: Opportunities and threats for sustainable ruminant systems.

The identification of crops that simultaneously contribute to the global protein supply and mitigate the effects of climate change is an urgent matter. Lupins are well adapted to nutrient-poor or contaminated soils, tolerate various abiotic stresses, and present relevant traits for acting as ecosystem engineers. Lupins are best studied for their seeds, but their full foraging potential needs further evaluation. This study evaluated the effects of location and sowing date on forage production, proximate composition, and the detailed mineral and alkaloid profiles of three species of Lupinus (L. albus cv. Estoril, L. angustifolius cv. Tango, and L. luteus cv. Cardiga). Sowing date and location and their interaction with the plant species significantly affected the vast majority of measured parameters, emphasizing the effects of climate and soil conditions on these crops. The relatively high crude protein and in vitro digestibility support the potential of the lupin species studied as sustainable forage protein sources in diets for ruminant animals. The content of individual essential macro and trace elements was below the maximum tolerable levels for cattle and sheep. Lupanine, smipine, and sparteine were the most abundant quinolizidine alkaloids in L. albus cv. Estoril, lupanine, and sparteine in L. angustifolius cv. Tango, and lupinine, gramine, ammodendrine, and sparteine in L. luteus cv. Cardiga. Based on the maximum tolerable levels of total quinolizidine alkaloid intake, the dietary inclusion of forages of L. albus cv. Estoril and L. angustifolius cv. Tango does not pose a risk to the animals, but the high alkaloid content of L. luteus cv. Cardiga may compromise its utilization at high levels in the diet. Overall, the results reveal a high potential for lupins as protein forage sources well adapted to temperate regions and soils with lower fertility, with a relevant impact on livestock sustainability in a climate change era.

A Preliminary Investigation of Salmonella Populations in Indigenous Portuguese Layer Hen Breeds.

The sustainability of agroecological systems, biodiversity protection, animal welfare, and consumer demand for higher quality products from alternative and extensive farming methods have reinforced interest in local breeds that are well adapted to low-input environments. However, food safety needs to be safeguarded to reinforce consumer confidence. The aim of this study was to conduct a preliminary investigation on the occurrence of Salmonella spp. in eggshells, hen's cloaca, and litter materials from autochthonous Portuguese laying hens raised in a semi-extensive system for small-scale production. A total of 279 samples from 31 flocks belonging to 12 farms were obtained, with 63 samples from the "Preta Lusitânica" breed, and 72 samples each from the remaining autochthonous breeds, namely, "Branca", "Amarela", and "Pedrês Portuguesa". None (0%) of the samples analyzed were positive for Salmonella spp. To the best of our knowledge, these are the first results of Salmonella evaluation from hen's cloaca, eggshells, and litter materials in autochthonous Portuguese chickens, suggesting that a semi-extensive production system can contribute to better food security and a lower risk to public health and the environment.

Antibiotic Resistance of Enterococcus Species in Ornamental Animal Feed.

Enterococcus is a bacterial genus that is strongly associated with nosocomial infections and has a high capacity to transfer and acquire resistance genes. In this study, the main objective was to evaluate the presence of Enterococcus species in ornamental animal feed and characterize their antimicrobial resistance and virulence factors. Antimicrobial susceptibility was determined using 14 antimicrobial agents by the disk diffusion method, complemented by genotypic analysis to identify Enterococcus species and the presence of 14 antimicrobial resistance and 10 virulence genes. From 57 samples of ornamental animal feed, 103 Enterococcus isolates were recovered from 15 bird, 9 fish and 4 reptile feed samples. Enterococcus isolates were highly resistance to rifampicin (78%) and erythromycin (48%), and 48% of isolates were classified as multidrug-resistant. Enterococcus faecalis (36.7%) and E. faecium (31.7%) were the species most frequently identified. Most isolates carried the resistance genes ermB (57%) and tetL (52%) and the virulence genes, cylL (52%) and esp (40%). Enterococcus gallinarum was the species with the highest number of multidrug-resistant isolates (50%) and virulence genes (80%). These results highlight the high levels of antibiotic-resistant Enterococcus spp. present in ornamental animal feed and the growing interaction of these animals with humans as a public health concern.

Microbiological Assessment of White Button Mushrooms with an Edible Film Coating.

The development of edible coatings incorporating bioextracts from mushrooms native to Portuguese forests aims to enhance the value of the endogenous forest and mycological resources by harnessing their potential as a source of antimicrobial and antioxidant compounds. Edible coatings represent an important pathway to decreasing food waste and contributing to implementing a circular bioeconomy. The coating should result in product valorization through improved preservation/conservation, increased shelf life, as well as enhancement of its antioxidant and enzymatic properties. To evaluate the effectiveness of an edible coating on fungal food matrices, a 14-day shelf-life study was conducted, wherein both coated and untreated mushrooms were examined under controlled storage temperatures of 4 °C and 9.3 °C. Agaricus bisporus was chosen as the food matrix for its bioeconomy significance, and Pleurotus eryngii was selected for the preparation of the food-based coating due to its profile of bioactive compounds. Microbiological analysis and physicochemical monitoring were conducted on the food matrices and the coating. Coated mushrooms had less mass loss and color change, and had better texture after 14 days. Microbiological analysis revealed that the coating had no antimicrobial activity. Overall, the coating improved the shelf life of the coated mushrooms but had less effect on the microbial community.

Upcycling post-harvest biomass residues from native European Lupinus species: from straws and pod shells production to nutritive value and alkaloids content for ruminant animals.

The production of Lupinus seeds for food and feed is increasing worldwide, which results in large amounts of post-harvest biomass residues, considered of low value and left in the field to be burned or incorporated in the soil. To valorize these agricultural wastes, this work aimed to assess their potential as an alternative feed for ruminants. Thus, the production yield, nutritive value, and alkaloid content of straws and pod shells from three native European Lupinus species, L. albus 'Estoril' (white), L. angustifolius 'Tango' (narrow-leafed), and L. luteus 'Cardiga' (yellow), cultivated in two locations, were evaluated. The dry matter (DM) yield of straws and pod shells were the highest for L. albus 'Estoril' (4.10 t ha-1) and the lowest for L. angustifolius 'Tango' (1.78 t ha-1), suggesting a poor adaptation of narrow-leafed lupin to the particularly dry and warm agronomic year. Despite species-specific differences, lupin biomass residues presented higher crude protein (53.0-68.9 g kg-1 DM) and lignin (103-111 g kg-1 DM) content than cereal straws usually used in ruminant feeding, thus resulting in higher metabolizable energy (6.43-6.58 MJ kg-1 DM) content. In vitro digestibility was similar among lupin species (47.7-50.6%) and higher in pod shells (53.7%) than in straws (44.6%). Lupinus albus 'Estoril' and L. luteus 'Cardiga' presented considerable amounts of alkaloids in straws (23.9 and 119 mg kg-1 DM) and pod shells (20.5 and 298 mg kg-1 DM), while no alkaloids were detected in L. angustifolius 'Tango' biomass residues. Considering the combined production of straw and pod shells per lupin species, it is anticipated that lupin biomass residues produced per ha can fulfill 85% of the energy and nearly 50% of protein requirements of a flock of 4 to 9 dry and mid-pregnancy sheep with 50 kg body weight for one year. No negative effects on small (ovine) and large (bovine) ruminant species due to alkaloids are expected, even if biomass residues are consumed at up to 85% DM intake. The large production yield along with its nutritive value unveils the potential of lupin biomass residues valorization as alternative fodder for ruminants, promoting sustainability under a circular economy approach.

Bovine Colostrum: Human and Animal Health Benefits or Route Transmission of Antibiotic Resistance-One Health Perspective.

After calving, bovine colostrum is obtained from the mammary gland of the dam in the first days and fed to newborn ruminant to prevent microbial infections. Each bovine colostrum has a unique biochemical composition with high nutraceutical value compared to milk. However, bovine colostrum is influenced by various factors, such as environmental, individual, and genetic factors, as well as processing methods. Proper colostrum management is crucial for obtaining high-quality colostrum and mitigating bacterial contamination. This is important not only for the health and survival of calves but also for the health of humans who consume colostrum and its co-products. It is essential to ensure that the consumed colostrum is free of pathogens to reap its benefits. Health-promoting products based on colostrum have gained significant interest. However, colostrum can contain pathogens that, if not eliminated, can contribute to their transmission and spread, as well as antibiotic resistance. The aim of this review was to promote the animal and human health benefits of bovine colostrum by improving its microbial quality and highlighting potential routes of dissemination of antibiotic-resistant pathogens. Implementing hygienic measures is one of the key factors in mitigating colostrum bacterial contamination and obtaining safe and high-quality colostrum. This helps reduce the exposure of pathogens to newborn calves, other animals, and humans, in a One Health analysis.

Amplifications of AURKA and AURKB in a Burkitt lymphoma immunodeficiency-associated type: a case report.

In equatorial Brazil, the association of Burkitt lymphoma and Epstein-Barr virus manifests at high rates. Here, we report, for the first time, amplifications of aurora kinase genes (AURKA/B) in a patient with a history of periodontal abscess and the presence of a remaining nodule, diagnosed with Burkitt lymphoma and Epstein-Barr virus, and /HIV positive. The patient was a 38-year-old man who presented with a 2-week-old severe jaw pain and a 3-day-old severe bilateral headache. He had a history of human papilloma virus. Interphase FISH analysis showed AURKA and AURKB amplification. The patient's condition worsened, progressing to death a month after the initial care. Changes in the MYCC and AURKA pathways are directly associated with genomic instability. Thus, MYCC rearrangements and higher expression of AURKA/B may be associated with therapy resistance, highlighting the importance of AURKA/B evaluation in Burkitt lymphoma.

Analysis of Antibiotic-Resistant and Virulence Genes of Enterococcus Detected in Calf Colostrum-One Health Perspective.

Enterococci are considered among the most prevalent global multidrug-resistant microorganisms globally. Their dissemination is a global concern, particularly by food-producing animals for both animals and humans. The aim of this study was to identify the species and investigate the antibiotic resistance and virulence profile of Enterococcus in bovine colostrum. Out of 88 presumptive Enterococcus isolates, species identification and susceptibility to 14 antimicrobials were tested using the disk diffusion method. An analysis of the antibiotic resistance and virulence genes was performed on the most prevalent species, using specific PCR assays. Enterococcus faecalis (54.5%), E. faecium (14.8%) and E. gallinarum (6.8%) were the identified species. To the best of our knowledge, this is the first report of E. gallinarum in bovine colostrum. The majority of the isolates showed resistance to quinupristin-dalfopristin (95.9%), erythromycin (80.7%), tetracycline (80.7%) and streptomycin (58%). Ninety-two percent of isolates were classified as multidrug-resistant. The most frequently detected resistance genes were tet(K) (61.1%), tet(M) (75.9%), tet(L) (90.7%), erm(B) (55.6%) and ant(6)-Ia (46.3%). The most prevalent virulence factors were cpd, esp, agg and cylLL. Enterococcus faecium showed a higher probability of carrying the erm(C), tet(M), ace and gel(E) genes (p < 0.05). These results demonstrated that colostrum can constitute an important reservoir and vehicle for the dissemination of antibiotic resistance and virulence genes to the three niches included in a One Health perspective (humans, animals and the environment), highlighting the importance of hygiene sanitary measures to mitigate colostrum microbial contamination.

Farm to table: colistin resistance hitchhiking through food.

Colistin is a high priority, last-resort antibiotic recklessly used in livestock and poultry farms. It is used as an antibiotic for treating multi-drug resistant Gram-negative bacterial infections as well as a growth promoter in poultry and animal farms. The sub-therapeutic doses of colistin exert a selection pressure on bacteria leading to the emergence of colistin resistance in the environment. Colistin resistance gene, mcr are mostly plasmid-mediated, amplifying the horizontal gene transfer. Food products such as chicken, meat, pork etc. disseminate colistin resistance to humans through zoonotic transfer. The antimicrobial residues used in livestock and poultry often leaches to soil and water through faeces. This review highlights the recent status of colistin use in food-producing animals, its association with colistin resistance adversely affecting public health. The underlying mechanism of colistin resistance has been explored. The prohibition of over-the-counter colistin sales and as growth promoters for animals and broilers has exhibited effective stewardship of colistin resistance in several countries.

Amplifications of AURKA and AURKB in a Burkitt lymphoma immunodeficiency-associated type: a case report

ABSTRACT In equatorial Brazil, the association of Burkitt lymphoma and Epstein-Barr virus manifests at high rates. Here, we report, for the first time, amplifications of aurora kinase genes (AURKA/B) in a patient with a history of periodontal abscess and the presence of a remaining nodule, diagnosed with Burkitt lymphoma and Epstein-Barr virus, and /HIV positive. The patient was a 38-year-old man who presented with a 2-week-old severe jaw pain and a 3-day-old severe bilateral headache. He had a history of human papilloma virus. Interphase FISH analysis showed AURKA and AURKB amplification. The patient's condition worsened, progressing to death a month after the initial care. Changes in the MYCC and AURKA pathways are directly associated with genomic instability. Thus, MYCC rearrangements and higher expression of AURKA/B may be associated with therapy resistance, highlighting the importance of AURKA/B evaluation in Burkitt lymphoma.

Insights from the yield, protein production, and detailed alkaloid composition of white (Lupinus albus), narrow-leafed (Lupinus angustifolius), and yellow (Lupinus luteus) lupin cultivars in the Mediterranean region.

Introduction: Lupins and other legumes have been considered as alternative plant-based protein sources to soybeans for both humans and livestock. Furthermore, they can contribute to more sustainable agricultural systems. The productivity and chemical composition of legumes is highly variable between species, cultivars, and with the edaphoclimatic conditions. Methods: This work evaluated the adaptability of seven Lupinus cultivars in two different sowing locations, during two consecutive years, through the characterization of their seed, as a means of investigating their suitability to be used as a source of food and/or feed. Results and discussion: Lupinus angustifolius cv. Tango and Lupinus luteus cv. Acos were the most stable genotypes across the environments when considering the seed and protein production, while L. luteus cv. Alburquerque and L. luteus cv. Mister showed less variation in the total alkaloid content across the environments. The edaphoclimatic conditions affected seed and protein yields, as higher rainfall resulted in high productivity. The lower temperatures observed in the first year at both locations caused a reduction in the production of alkaloids in L. luteus cv. Acos and Cardiga. Due to the high alkaloid content of some of the studied cultivars their use as food or feed can pose some safety concerns. However, these cultivars can have high levels of resistance to herbivore and insect attacks, which can be of the utmost importance for the use of these crops for recovering poor or exhausted soils.

Ammonia and greenhouse emissions from cow's excreta are affected by feeding system, stage of lactation and sampling time.

Decomposition of dairy cows' excreta on housing floor leads to ammonia and greenhouse gases production, yet factors affecting total emissions have not been fully disclosed. This work aimed to assess the impact of lactation stage, feeding system and sampling time on gaseous emission potential of cow's faeces and urine in laboratory chambers systems. Individual faeces and urine were collected from two groups of four cows, at peak and post peak lactation, from three commercial farms with distinct feeding systems: total mixed ration (TMR), total mixed ration plus concentrate at robot (TMR + robot), and total mixed ration plus concentrate in automatic feeders (TMR + AF). Samples were collected before a.m. (T8h), at middle day (T12h), and before p.m. (T17h) milking. In a laboratory chambers system, faeces and urine were mixed in a ratio of 1.7:1, and ammonia and greenhouse gases emissions were monitored during 48-h. Cumulative N-N2O emissions were the highest in TMR + robot system, post peak cows and sampling time T17h. An interaction between stage of lactation and sampling time was detected for N-NH3 and N-N2O (g/kg organic soluble N) emissions. Post peak cows also produced the highest cumulative N-NH3 emissions. Overall results contribute for the identification of specific on-farm strategies to reduce gaseous emissions from cows' excreta.

Immunoexpression of PD-L1, CD4+ and CD8+ cell infiltrates and tumor-infiltrating lymphocytes (TILs) in the microenvironment of actinic cheilitis and lower lip squamous cell carcinoma.

OBJECTIVE: Lower lip squamous cell carcinomas (LLSCC) could be associated with a previous history of potentially malignant oral diseases (PMOD), especially actinic cheilitis (AC), with high sun exposure being a well-described risk factor. Immune evasion mechanisms, such as the PD-1/PD-L1 (programmed cell death protein 1/programmed death-ligand 1) pathway has been gaining prominence since immunotherapy with immune checkpoint inhibitors showed a positive effect on the survival of patients with different types of neoplasms. Concomitant with the characterization of the tumor microenvironment, the expression of either or both PD-1 and PD-L1 molecules may estimate mutual relations of progression or regression of the carcinoma and prognostic values of the patient.Considering the importance of tumor microenvironment characterization, this study aims to determine the immunoexpression of PD-L1 and correlate with the frequency of CD4+ and CD8+ cells in AC and LLSCC lesions and with tumor-infiltrating lymphocytes (TILs) in LLSCC and its relationship with histopathological characteristics. METHODOLOGY: This sample includes 33 cases of AC and 17 cases of LLSCC. The cases were submitted to histopathological analysis and to CD4+, CD8+, and PD-L1+ cell determination by immunohistochemistry. RESULTS: There was a significant difference among the frequencies of CD4+, CD8+, and PD-L1+ cells between AC and LSCC cases, higher in the last group. Moreover, histopathological and atypical changes in AC and LLSCC were correlated with the frequencies of PD-L1+, CD4+, and CD8+ cells. In AC, PD-L1+ cases had a low frequency of CD4+ cells, but on the other hand, PD-L1+ cases of LLSCC had a higher frequency of CD4+ and CD8+ cells. CONCLUSION: Therefore, the PD-L1 molecule may be a potential escape route for the immune response in oral lesions, but the mechanisms differ between AC and LLSCC. Future studies related to immune evasion and immunotherapy in oral lesions should consider the analysis of inflammatory infiltrate and TILs.

Potential Histopathological and Immune Biomarkers in Malignant and Non-Malignant Oral Lesions.

Objectives: The presented case-control study was developed to characterize the clinical, histopathological and immunological profile of patients with traumatic injuries, benign neoplasms, potentially malignant oral disorders and malignant neoplasms of the oral cavity, in order to identify biomarkers of malignancy. Material and Methods: Clinical information was collected from dental records and several techniques were performed, including histopathological evaluation in sections stained with haematoxylin and eosin, immunohistochemistry for programmed death ligand-1 and measurement of serum levels of interferon-gamma, interleukin-6, -10 and -12. Statistical analysis was performed using IBM SPSS® Statistics software. Results: This study included 8 patients with traumatic injuries, 8 with benign neoplasms, 6 with potentially malignant oral disorders and 11 with malignant neoplasms. An association was observed between the classification of the lesion and smoking (P < 0.05), the size of the lesion (P < 0.05), the density of the inflammatory infiltrate (P < 0.001), the degree of dysplasia (P < 0.01) and programmed death ligand-1 expression (P < 0.01). Conclusions: Therefore, it is suggested that smoking, the size of the lesion, the inflammatory infiltrate and the programmed death ligand-1 expression can be considered potential biomarkers of oral malignancy.

[Analysis of root sealing using methylene blue in different endodontic obturation techniques. Literature review]. / Análisis del sellado radicular utilizando azul de metileno en distintas técnicas de obturación endodóntica. Revisión de la literatura.

One of the biggest drawbacks in endodontic filling is the problem of insufficient root sealing, due to the microleakage of bacteria into the canal. The objective of the research was to carry out a narrative review of the characteristics of the use of methylene blue as an evaluator of microleakage in studies that compared the quality of root sealing with different endodontic filling techniques. The research method used was literature review. Methylene blue was used in experimental in vitro studies, which compared filling techniques such as lateral condensation, cold injectable gutta-percha, thermomechanics, among others, the sealed dental pieces were immersed in a 1% methylene blue dye at 5% concentration. This procedure allowed reevaluating the samples in different periods of time. In conclusion, for endodontic filling penetration testing, methylene blue is an easy-to-use and reliable option.

Sarcoma fibromixoide de baixo grau em região paratesticular: um relato de caso / Low-grade fibromyxoid sarcoma in paratesticular area: a case report

Objetivos: relatar um caso raro de sarcoma fibromixoide de baixo grau (SFMBG) em uma localização incomum de modo a reforçar aspectos histopatológicos e imunoistoquímicos relevantes para o reconhecimento desta entidade e o adequado diagnóstico diferencial de massas paratesticulares. Relato de caso: homem de 20 anos, com massa escrotal à direita, cuja análise histopatológica demonstrou a presença de tecido fibroso com áreas mixoides e predominância de células fusiformes. A imunoistoquímica foi positiva para vimentina, com índice de Ki67 de 2%, e negativa para S100, CD-34, beta-catenina, desmina e miogenina. Conclusões: caso raro de SFMBG na região paratesticular que reforça a importância da histopatologia e da imunoistoquímica no diagnóstico desse tumor. Apesar da característica histológica benigna, o SFMBG apresenta altas taxas de recorrência e metástases, sendo essencial o seguimento do paciente.

Objectives: to report a rare case of low-grade fibromyxoid sarcoma (LGFMS) in an unusual location in order to reinforce histopathological and immunohistochemical aspects relevant to the recognition of this entity and the adequate differential diagnosis of paratesticular masses. Case report: 20-year-old man, with a right scrotal mass and histopathological analysis showing the presence of fibrous tissue with myxoid areas and a predominance of spindle cells. Immunohistochemistry was positive for vimentin, with a Ki67 index of 2%, and negative for S100, CD-34, beta-catenin, desmin and myogenin. Conclusions: rare case of LGFMS in the paratesticular region that reinforces the importance of histopathology and immunohistochemistry in the diagnosis of this tumor. Despite the benign histological characteristic, LGFMS has high rates of recurrence and metastasis, and patient follow-up is essential.