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.

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.

Genomic evolution of the human and animal coronavirus diseases.

Different coronaviruses have emerged due to their ability to infect, mutate and recombine multiple species and cell types, suggesting that these viruses will carry on to evolve and origin both veterinary and human diseases. So far, more than fifteen coronavirus-related diseases have been described in animals and seven in humans. Of which recently, a novel human betacoronavirus designated by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), an emerging zoonotic coronavirus is the causative agent of the coronavirus disease 2019. This virus emerged in China and spread rapidly worldwide. At the end of January 2020, the WHO declared the pandemic as a public health emergency of international concern. In this pandemic, the SARS-CoV-2 virus has infected more than 198 million people, with 4.2 million deaths worldwide (as of 2 August 2021). In the past two decades, this is the third betacoronavirus that has crossed the interspecies barrier from animals to infect humans and other animal species. The diseases caused mainly severe respiratory infections. The aim of this review is to summarize and provide an overview of the coronaviruses that can affect animals and humans and the diseases that ensue, as well as, its genomic relationship.

Environmental implications of stored cattle slurry treatment with sulphuric acid and biochar: A life cycle assessment approach.

With the increase of animal slurry produced from livestock production, the monitoring and mitigation of greenhouse gas (GHG) and ammonia (NH3) emissions represent a major issue. Life cycle assessment (LCA) has been used to evaluate the long-term environmental effects of applied strategies and technologies on cattle slurry management for mitigation of environmental harmful gases. This study was carried on two main aims: first, the effect of the addition of sulphuric acid (SA), biochar (SBi) or A + Bi to liquid cattle-slurry (treated systems) on gas emissions during storage compared to the untreated system (S) was investigated in a laboratory-controlled experiment; second, the environmental implications of each treated or untreated system were assessed through a LCA approach according to ISO 14040/44. Five CML 2001 impact categories were used: eutrophication potential (EP), acidification potential (AP), global warming potential (GWP), human toxicity potential (HTP) and Ozone Layer Depletion Potential (ODP). Comparisons were based on 1 ton fresh dairy cattle slurry. The environmental profile of untreated system showed lower efficiency in mitigation of total GHG and NH3 emissions (0.0312 and 0.0001 kg CO2-eq respectively), during storage period and greater impact on GWP and HTP categories. The electricity consumption in mechanical separation dominated the environmental impacts. From the three proposed treated systems, SA showed the highest efficiency on mitigation of gas emissions compared to the other treatments, reducing NH3, CH4 and CO2 emissions respectively in 61%, 98% and 15%, when compared to the SBi system. In all categories, acidified slurry also showed the lowest environmental impact relative to other treated systems and the benefit was more evident when the impacts were expressed per kilogram of nitrogen in the slurry. Therefore, LCA methodology was of great importance to assess the environmental implications of the treatments and acidification can be considered as an effective technique on the mitigation of environmental implications of livestock production and cattle-effluent valorization. Optimization and uniformity of performed studies are essential to validate new strategies to improve the sustainability of this sector in the management of animal wastewater.

Effects of chemical and physical methods on decellularization of murine skeletal muscles.

Volumetric muscle loss causes functional weakness and is often treated with muscle grafts or implant of biomaterials. Extracellular matrices, obtained through tissue decellularization, have been widely used as biological biomaterials in tissue engineering. Optimal decellularization method varies among tissues and have significant impact on the quality of the matrix. This study aimed at comparing the efficacy of four protocols, that varied according to the temperature of tissue storage and the sequence of chemical reagents, to decellularize murine skeletal muscles. Tibialis anterior muscles were harvested from rats and were frozen at -20°C or stored at room temperature, followed by decellularization in solutions containing EDTA + Tris, SDS and Triton X-100, applied in different sequences. Samples were analyzed for macroscopic aspects, cell removal, decrease of DNA content, preservation of proteins and three-dimensional structure of the matrices. Processing protocols that started with incubation in SDS solution optimized removal of cells and DNA content and preserved the matrix ultrastructure and composition, compared to those that were initiated with EDTA + Tris. Freezing the samples before decellularization favored cell removal, regardless of the sequence of chemical reagents. Thus, to freeze skeletal muscles and to start decellularization with 1% SDS solution showed the best results.

Implications of antibiotics use during the COVID-19 pandemic: present and future.

COVID-19 is caused by the SARS-CoV-2 virus, which has infected more than 4 million people with 278 892 deaths worldwide as of 11 May 2020. This disease, which can manifest as a severe respiratory infection, has been declared as a public health emergency of international concern and is being treated with a variety of antivirals, antibiotics and antifungals. This article highlights the administration of antimicrobials in COVID-19 patients worldwide, during the 2019-20 pandemic. It is imperative to be aware of the unreported amounts of antibiotics that have been administered worldwide in just a few months and a marked increase in antimicrobial resistance should therefore be expected. Due to the lack of data about antimicrobial use during this pandemic, the global impact on the emergence of new antimicrobial resistance is as yet unknown. This issue must be at the forefront of public health policymaking and planning in order that we are prepared for the potentially severe consequences for human and animal health and the environment.

A lower left atrial appendage peak emptying velocity in the acute phase of cryptogenic stroke predicts atrial fibrillation occurrence during follow-up.

BACKGROUND: Silent atrial fibrillation is a frequent etiology of cryptogenic stroke. Spontaneous conversion of atrial fibrillation to sinus rhythm results in atrial stunning. OBJECTIVE: To evaluate if the presence of a lower left atrial appendage peak emptying velocity (LAAV) after a cryptogenic stroke is associated with the occurrence of atrial fibrillation (AF). METHODS: We retrospectively selected consecutive patients with an acute ischemic stroke that had a transoesophageal echocardiogram (TEE) performed in the first 30 days of the acute event. Documented AF or potential cardioembolic sources in the TEE were considered exclusion criteria. We assessed the LAAV. During follow-up, we evaluated the occurrence of new-onset AF and the combined endpoint of death or new ischemic stroke. RESULTS: We studied 73 consecutive patients, during a mean follow-up period of 54.9 ± 19.3 months. Seven developed AF, and 13 had the combined endpoint. LAAV was independently associated with AF occurrence (HR: 0.93, 95% CI: 0.88-0.99; P = .016). Patients with a LAAV ≤ 46.5 cm/s (AUC: 0.766, 95% CI: 0.579-0.954; P = .021) had a lower survival rate free from AF occurrence (Log-rank, P < .001) and free from the combined endpoint of death or ischemic stroke (Log-rank, P = .010). CONCLUSION: A lower LAAV was associated with AF occurrence and the combined endpoint of death or ischemic stroke after an initial episode of cryptogenic stroke. Patients with this finding could eventually benefit from long-term cardiac rhythm monitoring.

Impact of different treatments on Escherichia coli during storage of cattle slurry.

There are different types of effluents from farming operations including untreated slurry (a mixture of manure, urine, split feed, and water), and treated slurry that normally is filtered to separate the solid fraction from the liquid fraction. With the amount of slurry applied on the soils as fertiliser every year, there are necessary to measure the leaching of microbial capable of transmitting infective agents that can be normally on slurry, because slurry can be a potential biohazard capable of transmitting infective agents. The aim of this study was to investigate the presence and survival of Escherichia coli (E. coli) on liquid fraction of dairy slurry, with the addition of different treatments during storage, such as addition of Biochar, beneficial microorganisms, sulphuric acid and the combinations of them. All the applied treatments to slurry show statistically significant differences (P < 0.001), according to the different sampling dates. Results showed that there are conditions and treatments that benefit the survival probability of E. coli, the treatments that include the acidification of slurry have the highest averages of CFU/ml (243288.3 at 37 °C; 136584 at 44 °C). These results can contribute to the improvement of the quality of slurry applied on soils, beneficiating agriculture but also public health.

Clinical applications of pharmacogenomics.

Pharmacogenomics is an emergent field aimed at tailoring pharmacological therapy. Genetic polymorphisms can modify the expression and function of enzymes and proteins involved in drug metabolism, affecting absorption, distribution, biotransformation and excretion as well as the drug-target interaction. Therefore, the presence of allelic variants will classify people as poor, extensive or rapid/ultra rapid metabolizers, modifying drug efficacy and safety. In this work, the state of art in relation to this discipline is presented and the genetic variants of enzymes that are involved in drug pharmacokinetics or pharmacodynamics are described. The effects of these variants on the therapeutic response to drugs used in our country are also discussed.

Canine parvovirus: the worldwide occurrence of antigenic variants.

The most important enteric virus infecting canids is canine parvovirus type 2 (CPV-2). CPV is the aetiologic agent of a contagious disease, mainly characterized by clinical gastroenteritis signs in younger dogs. CPV-2 emerged as a new virus in the late 1970s, which could infect domestic dogs, and became distributed in the global dog population within 2 years. A few years later, the virus's original type was replaced by a new genetic and antigenic variant, called CPV-2a. Around 1984 and 2000, virus variants with the single change to Asp or Glu in the VP2 residue 426 were detected (sometimes termed CPV-2b and -2c). The genetic and antigenic changes in the variants have also been correlated with changes in their host range; in particular, in the ability to replicate in cats and also host range differences in canine and other tissue culture cells. CPV-2 variants have been circulating among wild carnivores and have been well-documented in several countries around the world. Here, we have reviewed and summarized the current information about the worldwide distribution and evolution of CPV-2 variants since they emerged, as well as the host ranges they are associated with.

The Placenta as an Organ and a Source of Stem Cells and Extracellular Matrix: A Review.

The placenta is a temporal, dynamic and diverse organ with important immunological features that facilitate embryonic and fetal development and survival, notwithstanding the fact that several aspects of its formation and function closely resemble tumor progression. Placentation in mammals is commonly used to characterize the evolution of species, including insights into human evolution. Although most placentas are discarded after birth, they are a high-yield source for the isolation of stem/progenitor cells and are rich in extracellular matrix (ECM), representing an important resource for regenerative medicine purposes. Interactions among cells, ECM and bioactive molecules regulate tissue and organ generation and comprise the foundation of tissue engineering. In the present article, differences among several mammalian species regarding the placental types and classifications, phenotypes and potency of placenta-derived stem/progenitor cells, placental ECM components and current placental ECM applications were reviewed to highlight their potential clinical and biomedical relevance.

Porcine hokovirus in wild boar in Portugal.

Porcine hokovirus (PHoV), also referred to as porcine parvovirus 4 (P-PARV4), a recently discovered parvovirus of swine that is closely related to human parvovirus 4/5 (H-PARV4/5), was first described in Hong Kong. To evaluate the occurrence of P-PARV4 in Portuguese wild boars in the hunting season of 2011/2012, liver and serum samples were tested. P-PARV4 was detected in 24 % of the wild boars analyzed. Phylogenetic analysis showed a close relationship between the P-PARV4 isolates and other P-PARV4 reference strains. This virus appears to be emerging, with yet unknown implications for public health.

Characterization of the CYP2D6 drug metabolizing phenotypes of the Chilean mestizo population through polymorphism analyses.

We tested the influence of four polymorphisms and gene duplication in CYP2D6 on in vivo enzyme activity in a Chilean mestizo population in order to identify the most relevant genetic profiles that account for observed phenotypes in this ethnic group. CYP2D6*2 (2850C>T), *3 (2549A>del), *4 (1846G>A), *17 (1023C>T) and gene duplication were determined by PCR-RFLP or PCRL in a group of 321 healthy volunteers. Individuals with different variant alleles were phenotyped by determining debrisoquine 4-hydroxylase activity as a metabolic ratio (MR) using a validated HPLC assay. Minor allele frequencies were 0.41, 0.01, 0.12 and 0.00 for CYP2D6*2, *3, *4 and *17 variants, respectively, and the duplication frequency was 0.003. Genotype analysis correlated with phenotypes in 18 of 23 subjects (78.3%). 11 subjects were extensive metabolizers (EM), 8 were intermediate metabolizers (IM), 2 were poor metabolizers (PM) and 2 were ultra-rapid metabolizers (UM) which is fairly coincident with expected phenotypes metabolic ratios ranged from 0.11 to 126.41. The influence of CYP2D6*3 was particularly notable, although only heterozygote carriers were present in our population. Individuals homozygous for *4 were always PM. As expected, the only subject with gene duplication was UM. In conclusion, there was a clear effect of genotype on observed CYP2D6 activity. Classification of EM, PM and UM through genotyping was useful to characterize CYP2D6 phenotype in the Chilean mestizo population.

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.

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.

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.

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.

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.