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This study aimed to count Enterobacteriaceae and Escherichia coli in different locations on pig carcasses (shank, loin, abdomen, shoulder, and jowl) from two slaughterhouses (A and B) between September 2019 and July 2021 during different slaughter stages (after bleeding, after passing through the epilator machine, after manual toileting in the dirty area, before and after evisceration, and after the final washing), as well as verify antimicrobial resistance and biofilm formation capacity. The main points of Enterobacteriaceae and E. coli contamination were identified in the two slaughterhouses through three collections. The stages with the highest counts were post-bleeding and evisceration in both slaughterhouses and after manual toileting in slaughterhouse B in the first collection. Most E. coli isolates were resistant to multiple antimicrobials, with higher resistance frequencies to amoxicillin, ampicillin, chloramphenicol, sulfonamides, and streptomycin. The virulence genes eae , stx1 , and stx2 were also detected. Three isolates had all three genes and exhibited resistance to at least six antimicrobial classes (β-lactams, macrolides, aminoglycosides, sulfonamides, amphenicols, and quinolones). E. coli isolates also showed a high frequency of strains with moderate and strong in vitro biofilm-forming capacity. This is the first study to characterize microbial contamination by pig slaughter stage in the Federal District region, demonstrating the critical points for hygienic production. E. coli was isolated from the surface of pig carcasses, as well as the virulence genes stx1 , stx2 , and eae were detected. The multi-antimicrobial resistant isolates also had a moderate-to-strong biofilm formation capacity, thus demonstrating risks to public health.

Background and Aims: Ovum pick-up (OPU) is an intrinsic step of in vitro fertilization procedures. Nevertheless, it can cause ovarian lesions and compromise female fertility in bovines. Recently, we have shown that intraovarian injection of adipose-derived mesenchymal stromal cells (AD-MSCs) effectively preserves ovarian function in bovines. Given that MSC-derived extracellular vesicles (MSC-EVs) have been shown to recapitulate several therapeutic effects attributed to AD-MSCs and that they present logistic and regulatory advantages compared to AD-MSCs, we tested whether MSC-EVs would also be useful to treat OPU-induced lesions., Methods: MSC-EVs were isolated from the secretome of bovine AD-MSCs, using ultrafiltration (UF) and ultracentrifugation methods. The MSC-EVs were characterized according to concentration and mean particle size, morphology, protein concentration and EV markers, miRNA, mRNA, long noncoding RNA profile, total RNA yield and potential for induction of the proliferation and migration of bovine ovarian stromal cells. We then investigated whether intraovarian injection of MSC-EVs obtained by UF would reduce the negative effects of acute OPU-induced ovarian lesions in bovines. To do so, 20 animals were divided into 4 experimental groups (n = 5), submitted to 4 OPU cycles and different experimental treatments including vehicle only (G1), MSC-EVs produced by 7.5 × 10 6 AD-MSCs (G2), MSC-EVs produced by 2.5 × 10 6 AD-MSCs (G3) or 3 doses of MSC-EVs produced by 2.5 × 10 6 AD-MSCs, injected after OPU sessions 1, 2 and 3 (G4)., Results: Characterization of the MSC-EVs revealed that the size of the particles was similar in the different isolation methods; however, the UF method generated a greater MSC-EV yield. MSC-EVs processed by both methods demonstrated a similar ability to promote cell migration and proliferation in ovarian stromal cells. Considering the higher yield and lower complexity of the UF method, UF-MSC-EVs were used in the in vivo experiment. We evaluated three therapeutic regimens for cows subjected to OPU, noting that the group treated with three MSC-EV injections (G4) maintained oocyte production and increased in vitro embryo production, compared to G1, which presented compromised embryo production following the OPU-induced lesions., Conclusions: MSC-EVs have beneficial effects both on the migration and proliferation of ovarian stromal cells and on the fertility of bovines with follicular puncture injury in vivo., Competing Interests: Declaration of competing interest Authors MP and PM declare competing financial interests as owners of the company Bio. All other authors declare that they have no conflicting interests., (Copyright © 2024 International Society for Cell & Gene Therapy. Published by Elsevier Inc. All rights reserved.)

Introduction:  The use of mesenchymal stem cells (MSC) in cytotoxicity tests is an in-vitro alternative model for predicting initial doses. Homeopathic medicines may stimulate the immune system to combat a pathology effectively and have been used for over two centuries. Viscum album (VA) extracts are widely used in the treatment of cancer, due to their immunomodulatory, cytotoxic and pro-apoptotic properties., Objective:  This study aimed to evaluate the in-vitro growth kinetics of canine MSC in relation to cytotoxicity, cell differentiation and expression of pluripotentiality markers, using a VA preparation at the D1D2 (1×10 -1 , 1×10 -2 potency (VAD1D2)., Methods:  MSC were obtained from adipose tissue sampled from a healthy dog that was undergoing an elective veterinary procedure and with its owner's permission. The experiments were performed in three groups: MSC treated with VAD1D2 or diluent or untreated (control). The cytotoxicity was evaluated by MTT assay. The differentiation was induced in three lineages, and apoptotic cell labeling was performed by an Annexin-V test., Results:  At the concentration of 10 μL/mL of VA, the number of cells after in-vitro culture was maintained when compared with the control (untreated) group. A significant and gradual decrease in cell viability was recorded as VA concentrations increased. The apoptosis analysis showed that VA at 20 μL/mL presented absolute percentages of initial apoptosis twice as high as at 10 μL/mL, which was similar to the control (untreated group)., Conclusion:  The results suggest that the use of efficient methods to assess the in-vitro cytotoxicity of VA-based homeopathic medicines using MSC lineages may predict the potential action at different concentrations. These findings demonstrated that VAD1D2 interferes with canine MSC growth kinetics., Competing Interests: None declared., (The Faculty of Homeopathy. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/).)

This study aimed to verify the presence of Listeria monocytogenes, Salmonella spp., and Escherichia coli in two Brazilian swine slaughterhouses, as well as to perform antibiograms, detect virulence and antimicrobial resistance genes, and evaluate the in vitro biofilm-forming capability of bacterial isolates from these environments. One Salmonella Typhi isolate and 21 E. coli isolates were detected, while L. monocytogenes was not detected. S. Typhi was isolated from the carcass cooling chamber's floor, resistant to several antimicrobials, including nalidixic acid, cefazolin, chloramphenicol, doxycycline, streptomycin, gentamicin, tetracycline, and sulfonamide, and contained resistance genes, such as tet(B), tet(C), tet(M), and ampC. It also showed moderate biofilm-forming capacity at 37°C after incubating for 72 h. The prevalence of the 21 E. coli isolates was also the highest on the carcass cooling chamber floor (three of the four samplings [75%]). The E. coli isolates were resistant to 12 of the 13 tested antimicrobials, and none showed sensitivity to chloramphenicol, an antimicrobial prohibited in animal feed since 2003 in Brazil. The resistance genes MCR-1, MCR-3, sul1, ampC, clmA, cat1, tet(A), tet(B), and blaSHV, as well as the virulence genes stx-1, hlyA, eae, tir α, tir β, tir γ, and saa were detected in the E. coli isolates. Moreover, 5 (23.8%) and 15 (71.4%) E. coli isolates presented strong and moderate biofilm-forming capacity, respectively. In general, the biofilm-forming capacity increased after incubating for 72 h at 10°C. The biofilm-forming capacity was the lowest after incubating for 24 h at 37°C. Due to the presence of resistance and virulence genes, multi-antimicrobial resistance, and biofilm-forming capacity, the results of this study suggest a risk to the public health as these pathogens are associated with foodborne diseases, which emphasizes the hazard of resistance gene propagation in the environment., Competing Interests: The authors have declared that no competing interests exist.

Listeria monocytogenes and Salmonella spp. are considered important foodborne pathogens that are commonly associated with foods of animal origin. The aim of this study was to perform molecular characterization of L. monocytogenes and Salmonella spp. isolated from biofilms of cattle and poultry slaughterhouses located in the Federal District and State of Goiás, Brazil. Fourteen L. monocytogenes isolates and one Salmonella sp. were detected in poultry slaughterhouses. No isolates were detected in cattle slaughterhouses. All L. monocytogenes isolates belonged to lineage II, and 11 different pulsotypes were detected. Pulsed-field gel electrophoresis analysis revealed the dissemination of two strains within one plant, in addition to the regional dissemination of one of them. The Salmonella isolate was identified via whole genome sequencing as Salmonella enterica serovar Minnesota ST548. In the sequence analysis, no premature stop codons were detected in the inlA gene of Listeria. All isolates demonstrated the ability to adhere to Caco-2 cells, while 50% were capable of invading them. Antimicrobial resistance was detected in 57.1% of the L. monocytogenes isolates, and resistance to sulfonamide was the most common feature. The tetC, ermB, and tetM genes were detected, and four isolates were classified as multidrug-resistant. Salmonella sp. was resistant to nine antimicrobials and was classified as multidrug-resistant. Resistance genes qnrB19, blaCMY-2, aac(6')-Iaa, sul2, and tetA, and a mutation in the parC gene were detected. The majority (78.5%) of the L. monocytogenes isolates were capable of forming biofilms after incubation at 37°C for 24 h, and 64.3% were capable of forming biofilms after incubation at 12°C for 168 h. There was no statistical difference in the biofilm-forming capacity under the different evaluated conditions. Salmonella sp. was capable of forming biofilms at both tested temperatures. Biofilm characterization was confirmed by collecting the samples consistently, at the same sampling points, and by assessing biofilm formation in vitro. These results highlight the potential risk of cross-contamination in poultry slaughterhouses and the importance of surveillance and pathogen control maintenance programs within the meat production industry., Competing Interests: The authors have declared that no competing interests exist.

Originated in Wuhan, China, the coronavirus 19 disease (COVID-19) has quickly spread worldwide, reaching countries that already faced other endemics and epidemics. In Brazil, such a concerning situation includes arboviruses, among which the dengue virus stands out. Here, we determined the rate of SARS-CoV-2/dengue virus co-infection in a total of 178 patients with COVID-19 symtoms admitted into a large public hospital of the Federal District of Brazil. Furthermore, we evaluated whether prior or active dengue virus infection influenced hematological, biochemical, and clinical parameters of such patients. One hundred and twelve (63%) individuals tested positive for COVID-19, of which 43 (38.4%) were co-infected with dengue virus, and 50 (44.6%) had antibodies indicative of previous dengue infection. Co-infected patients showed lower numbers of circulating lymphocytes and monocytes, higher glucose rates, and a worse pulmonary condition. Of note, prior infections with dengue virus did not influence clinical parameters, but active dengue fever resulted in higher hospitalization rate. In conclusion, amid the current complex epidemiological scenario in Brazil, our data support the notion that SARS-CoV-2 and dengue co-infection affects an important percentage of COVID-19 patients and leads to worse clinical parameters, requiring greater attention from health authorities., (Copyright © 2020. Published by Elsevier B.V.)

There are few animal germplasm/gene bank collections in Brazil, and basic studies are needed to attend the future internal and external demands from international partners. The aim of this work was to validate a "proof of concept" that integrates spatial (georeferenced data) and genetic data regarding the local of origin from 3518 DNA samples from 17 different genetic groups or breeds of sheep in the Brazilian Germplasm bank. Spatialisation shows that not all genetic groups have samples in the bank, and collection is concentrated in the conservation nuclei spread nationwide. Only 21% of states with a specific breed have samples in the gene bank. The mean number of animals sampled per collection was 32, while the mean distance travelled to collect samples was 262 km from the conservation nuclei. For example, the Brazilian Somali were only collected in the conservation nucleus in Ceará State. No samples were collected to date for the Cariri breed, which is recognised by the Brazilian Ministry of Agriculture. Only two farms and one breed in the bank are from the northern region. Of the 27 states, there are samples in the gene bank of sheep from 13, so several states have no samples, requiring collection from herds outside the official system of conservation to make sure that studies using this germplasm realised are not biased. Significant genetic differences are seen above 332 km, which should guide future sampling efforts. Suggestions are given for improving the quantity, quality and diversity of samples in the gene bank.

The goal of this research was to evaluate the microclimate (temperature, relative humidity and ECI-enthalpy comfort index) of commercial loads of broiler chickens at different transport distances: Dist15 (15 km on average) and Dist90 (90 km on average) in the summer and winter seasons and their effects on the production parameters body weight difference (BWD), mortality (%) and bruising prevalence (%). Twelve broiler loads were monitored using dataloggers to record temperature and humidity, with a total of 24 target crates per load. The experiment followed a factorial design [2 seasons (rainy and dry) × 2 distances (Dist15 and Dist90)] with a randomized complete block arrangement, 3 sexes (all males, all females, or mixed shipments) and one shipment per combination. BWD had a heterogeneous distribution throughout the load, and this distribution was not significantly correlated with the mean ECI measured during transport at 12 positions along the load. In terms of comfort, summer is the most critical period for broiler transport. In the interaction between rainy season and Dist90, the highest ECI was scored in the lethal zone (where physiological mechanisms are not enough to control body temperature). Mortality during the rainy season was not significantly different between distances. However, during the dry season, mortality was twice as high as broilers that travelled for 15 km. The prevalence of bruising on carcasses was not affected by the interaction between season and distance. As we know, broiler chicken performance, during transport, can be also related to road conditions, being hard to evaluate the real impact of seasons and distances on animal welfare. Load microclimate can compromise broiler chicken welfare during transport and it does not necessary reflect significant losses pre and post-slaughter., Competing Interests: The authors have declared that no competing interests exist.

Image 1., (© 2019 Center for Food and Biomolecules, National Taiwan University. Production and hosting by Elsevier Taiwan LLC.)

Pre-loading handling and conditions of transport are related to welfare, disease risk and product quality of production animals. These steps continue to be one of the major animal management problems in Brazil. This study evaluated the effects of different types of pre-loading handling and road transport times on the haematological and biochemical traits of cattle. Eighteen male cattle were submitted to three travel times (24, 48 and 72 h) in a truck soon after load using different types of pre-loading handling: traditional (rough handling), training (gentle handling) and use of flags to movement cattle. Haematological traits, blood biochemical measures as well as blood and faecal cortisol were analysed in order to assess animal welfare and physiological status. The traditional management showed to be more stressful, also had animals with a greater number of neutrophils and lower numbers of lymphocytes than handling with flags, showing that animals submitted to more stressful situations can have compromised immune system. Serum aspartate aminotransferase concentrations were within the reference levels and when taken together with increased creatine kinase patterns observed indicate muscle damage in traditional management. Decrease in glucose concentrations over time from traditional management to flag management was observed, while fructosamine was increased in traditional management with 72 h of travel. When taken together, all reported factors, immune, enzymatic, energetic and hormonal, indicate that the quality of pre-loading handling and time of transport were determinant for animal welfare, its homeostatic balance and sanitary conditions.

This research aims to evaluate the microclimate of commercial loads of broiler chickens at different distances in the summer (rainy) and winter (dry) seasons and their effects on meat quality. Twelve broiler loads were monitored with a total of 24 crates per load. Data loggers were used to record temperature and relative humidity. The experiment followed a completely randomized design with 48 treatments in a factorial scheme (2 seasons: rainy and dry) x 2 (distances: short and long) x 12 (positions), with 3 replicates per experimental group. In the rainy season, meat quality was influenced by transport distance. For longer distances, it recorded the highest enthalpy comfort index (ECI), suggesting a tendency of dark, firm, and dry meat (DFD-like) and lower cooking losses (CL). The lowest ECI was recorded during the transport in dry season. Broiler chickens transported and slaughtered in the winter presented meat pH and L* (lightness) classified as "normal," but with higher cooking losses. For the shear force (SF), the seasons and distances had no significant influence on tenderness of the meat. Regarding the crate positioning in the load, no effect was observed during transport on this variable, given the meat quality characteristics of pH, L*, CL, and SF. These results suggest that the distance and the seasons present more influence on broiler meat quality than crate position in the truck., (© 2017 Poultry Science Association Inc.)