Antimicrobial (ESBL) resistance genes in faecal E. coli of calves fed waste milk with antimicrobial residues.

This research paper aimed to evaluate the association between feeding waste milk to calves and the occurrence of antimicrobial multi-resistance by extended spectrum ß-lactamase (ESBL) enzymes through determining their production by E. coli isolates from 32 dairy farms. Among ß-lactamase enzymes, ESBL provide resistance to a wide variety of ß-lactam antimicrobials including penicillin and 2nd, 3rd and 4th generation cephalosporins. Feeding waste milk to calves has been observed to lead to increased antimicrobial resistance in faecal isolates of calves. In each farm included in this study, faecal samples were collected from the rectum of five healthy calves in the first month of life and pooled into a single container. Five isolates from each pool were selected and confirmed to be E. coli by amplification of the 16S rRNA gene. ESBL production was confirmed phenotypically on 148 isolates from 31 farms by use of the double-disk synergy test. Genotypic confirmation of ESBL production was performed by PCR for the genes blaCTX-M-1, -2, -8, -9 and blaCMY-2. A questionnaire was also performed and a mixed logistic regression model was used to identify risk factors for the occurrence of antimicrobial resistance. A negative binomial regression model was also used, in order to assess whether there was any association between certain farm management practices and the number of ESBL-producing E. coli isolates from each farm. Phenotypic confirmation of ESBL production was obtained on 40 E. coli isolates from 15 farms (48.4%), whereas genotypic confirmation was obtained on 55 isolates from 20 farms (64.5%). The use of three or more different intramammary antimicrobials to treat mastitis within the previous year significantly impacted the number of ESBL-producing E. coli isolates; on farms that did so, there were more isolates in which ESBL-producing E. coli was present, when compared to farms that had used less formulations within the same time span.

A polymicrobial biofilm model for testing the antimicrobial potential of a nisin-biogel for canine periodontal disease control.

BACKGROUND: Periodontal disease (PD) in dogs is prompted by the establishment of a polymicrobial biofilm at the tooth surface and a subsequent host inflammatory response. Several strategies may be used for PD control, including dental hygiene home care procedures, like toothbrushing, special diet and chew toys that reduce dental plaque accumulation, or professional periodontal treatments. Aiming at PD control, a biogel composed by nisin and guar-gum was previously developed. This work aimed to establish an in vitro model mimicking the PD-associated biofilms and to evaluate the nisin-biogel inhibitory activity against this polymicrobial biofilm by determining its Minimum Biofilm Inhibitory (MBIC) and Eradication Concentrations (MBEC). Bacterial species tested included Neisseria zoodegmatis CCUG 52598T, Corynebacterium canis CCUG 58627T, Porphyromonas cangingivalis DSMZ VPB 4874, Peptostreptococcus canis CCUG 57081 and an Enterococcus faecalis isolate belonging to a collection of oral bacteria obtained from dogs with PD. Before establishing the biofilm, coaggregation between species was determined by optical density measurement after 2 and 24 hours. Nisin-biogel MBIC and MBEC values regarding the polymicrobial biofilm were determined using a modified version of the Calgary biofilm pin lid device, after confirming the presence of the five bacterial species by Fluorescent In Situ Hybridization. RESULTS: Only 40% of the bacterial dual suspensions were able to coaggregate at 2 hours, but all species tested exhibited a coaggregation percentage higher than 30% at 24 hours. It was possible to establish a 48 h polymicrobial biofilm model composed by the five bacterial species selected. This model was used to determine nisin-biogel MBIC (26.39 ± 5.89 µg/mL) and MBEC (62.5 ± 27.73 µg/mL) values. CONCLUSIONS: Our results showed that the nisin-biogel can inhibit and eradicate PD multispecies biofilms. As this in vitro model mimics an in vivo periodontal polymicrobial biofilm, our results reinforce the potential of the application of nisin-biogel for canine PD control.

Skin asepsis protocols as a preventive measure of surgical site infections in dogs: chlorhexidine-alcohol versus povidone-iodine.

BACKGROUND: Most of surgical site infections (SSI) are caused by commensal and pathogenic agents from the patient's microbiota, which may include antibiotic resistant strains. Pre-surgical asepsis of the skin is one of the preventive measures performed to reduce SSI incidence and also antibiotic resistance dissemination. However, in veterinary medicine there is no agreement on which biocide is the most effective. The aim of this study was to evaluate the effectiveness of two pre-surgical skin asepsis protocols in dogs. A total of 46 animals were randomly assigned for an asepsis protocol with an aqueous solution of 7.5% povidone-iodine or with an alcoholic solution of 2% chlorhexidine. For each dog, two skin swab samples were collected at pre-asepsis and post-asepsis, for bacterial quantification by conventional techniques and isolation of methicillin-resistant species. RESULTS: Most samples collected at the post-asepsis did not present bacterial growth, both for the animals subjected to the povidone-iodine (74%) or to the chlorhexidine (70%) protocols. In only 9% of the cases a significant bacterial logarithmic reduction was not observed, indicating possible resistance to these agents. Also, the logarithmic reduction of the bacterial quantification from pre- and post-asepsis time, was not statistically different for povidone-iodine (6.51 ± 1.94 log10) and chlorhexidine (6.46 ± 2.62 log10) protocol. From the 39% pre-asepsis swabs which showed bacterial growth in MRSA modified chromogenic agar medium, only one isolate was identified as Staphylococcus aureus and one as S. epidermidis. False positives were mainly other staphylococci species, as well as Enterobacteriaceae. CONCLUSIONS: Pre-surgical skin asepsis protocols with povidone-iodine or chlorhexidine showed similar efficacy in the elimination of methicillin resistant bacteria and preventing surgical site infections in dogs undergoing surgery.

Evaluation of Amazon fruits: chemical and nutritional studies on Borojoa sorbilis.

BACKGROUND: Borojoa sorbilis (Ducke) is an Amazonian species with edible fruits that are widely consumed by the local population, but little studied and not yet economically explored. Thus the aim of this study was to describe the chemical composition, volatile compounds, nutritional aspects and antioxidant activity of the fruit pulp and peel of B. sorbilis. RESULTS: Headspace solid-phase microextraction, using polydimethylsiloxane-divinylbenzene (PDMS-DVB) fiber, resulted in the identification of 59 substances in the pulp and peel of B. sorbilis fruits after analysis of the chromatograms obtained by gas chromatography-mass spectrometry using polar and nonpolar columns. Esters were the most abundant. Moisture, lipids, protein, dietary fiber, ash, carbohydrate, total energy value, titratable acid, soluble solids and pH were measured. Protein amount, fat content and antioxidant activity were low in both pulp and peel. Carbohydrate content was 179.2 and 134.9 g kg-1 in pulp and peel, respectively. CONCLUSION: This is the first report on the chemical characteristics, flavor and nutritional aspects of B. sorbilis fruit, which is essential to its economic exploitation. The high energy value associated with the carbohydrate content, plus the low fat content, contribute to a possible use of B. sorbilis fruits as a food supplement. © 2018 Society of Chemical Industry.

In Situ Cytokine Expression and Morphometric Evaluation of Total Collagen and Collagens Type I and Type III in Keloid Scars.

Keloids are characterized by excessive collagen deposition and growth beyond the edges of the initial injury, and cytokines may be related to their formation. The objective of this study was to evaluate the collagen fibers, analyze in situ expression of cytokines in keloid lesions, and compare to the control group. Results showed that there was a predominance of women and nonwhite and direct black ancestry. Keloid showed a significant increase in total and type III collagen. Significantly, the expression of mRNA for TGF-ß in keloid was increased, the expressions of IFN-γ, IFN-γR1, and IL-10 were lower, and IFN-γR1 and TNF-α had no statistical difference. Correlations between collagen type III and TGF-ß mRNA expression were positive and significant, IFN-γ, IFN-γR1, and IL-10 were negative and significant, and TNF-α showed no statistical difference. We conclude that there was a significant increase of total collagen in keloid and predominance of collagen type III compared to the controls, showing keloid as an immature lesion. There is a significant increase in TGF-ß mRNA in keloid lesions, and a significant decrease in IFN-γ and IL-10, suggesting that these cytokines are related to keloid lesions.

Semi-quantitative method for Staphylococci magnetic detection in raw milk.

Bovine mastitis is the most costly disease for dairy farmers, hence, control measures to prevent it are crucial for dairy farm sustainability. Staphylococcus aureus is considered a major mastitis pathogen because of its impact on milk quality and low cure rates. Prevention of S. aureus mastitis includes segregation of infected animals, whilst treatment of such animals should be performed for a longer time to improve cure rates. This makes identification of S. aureus infected quarters and animals of significant importance. The experiments reported in this research paper aimed to develop and validate a sensitive method for magnetic detection of S. aureus and of the Staphylococcus genus in raw milk samples. Mastitic milk samples were collected aseptically from 47 cows with subclinical mastitis, from 12 Portuguese dairy farms. Forty nine quarter milk samples were selected based on bacteriological results. All samples were submitted to PCR analysis. In parallel, these milk samples were mixed with a solution combining specific antibodies and magnetic nanoparticles, to be analysed using a lab-on-a-chip magnetoresistive cytometer, with microfluidic sample handling. The antibodies used in this work were a rabbit polyclonal IgG anti-S. aureus ScpA protein and a mouse monoclonal IgM anti-S. aureus ATCC 29740. This paper describes the methodology used for magnetic detection of bacteria, including analysis of false positive/negative results. This immunological recognition was able to detect bacterial presence above 100 cfu/ml, independently of antibody and targeted bacteria used in this work. Comparison with PCR results showed sensitivities of 57·1 and 79·3%, specificity values of 75 and 50%, and PPV values of 40 and 95·8% for magnetic identification of Staphylococci species with an anti-S. aureus antibody and an anti-Staphylococcus spp. antibody, respectively. Some constraints are described as well as the method's limitations in bacterial quantification. Sensitivities and specificities require to be improved, nevertheless, the methodology described may form the basis for a means of identifying S. aureus infected cows at the point of care.

Colorectal surgical mortality and morbidity in elderly patients: comparison of POSSUM, P-POSSUM, CR-POSSUM, and CR-BHOM.

PURPOSE: This study aims to compare the predictive value of POSSUM, P-POSSUM, CR-POSSUM and CR-BHOM in colorectal surgical mortality and morbidity in patients over 80 years old. METHODS: This is a retrospective observational longitudinal study. A total of 991 patients who underwent major colorectal surgery between 2008 and 2012 in a secondary hospital in Portugal were screened, and 204 who were over 80 years old were included. Subgroup analysis was performed for malignant/benign disease and emergent/elective surgery. The main outcome measure was 30-day postoperative mortality and morbidity with Clavien-Dindo classification ≥ 2. RESULTS: Of the 204 patients included in this study, 155 had malignant disease, and 65 underwent emergent procedures. Overall average age was 84.3 ± 3.9 years (range 80-100). Overall surgical mortality and morbidity were 18.6% (n = 38) and 52.4% (n = 87), respectively. Expected mortality followed the order P-POSSUM

Seasonal variation in trace and minor elements in Brazilian honey by total reflection X-ray fluorescence.

Honey is used as an alternative medicine and is a constituent of a healthy diet worldwide. Its composition is associated with botanical origin and, to some extent, geographical origin because soil and climate characteristics determine the melliferous flora. Also, the elements content in honey samples could give an indication of environmental pollution or geographical origin. The main purpose of this study was to evaluate seasonal patterns of essential elements of Brazilian honey. Honey was collected during spring, summer, autumn, and winter for 2 years to quantify K, Ca, Ti, Cr, Mn, Fe, Ni, Cu, Zn, Se, Br, and Sr using total reflection X-ray fluorescence spectroscopy (TXRF). Our results indicate no seasonal differences in concentration of Cr, Ni, Se, and Ti, although there were significant seasonal patterns in the composition of essential elements in honey, with higher concentrations of minor and trace elements, especially K and Ca of samples collected in spring and summer.

Dynamics of bovine intramammary infections due to coagulase-negative staphylococci on four farms.

The objectives of this study were to compare the impact of different coagulase-negative species (CNS) on udder health measured in terms of individual quarter milk somatic cell count (SCC) and duration of intramammary infection, and to get some insight into most likely routes of infection for different CNS species. This longitudinal observational study was performed on four farms that were sampled at 4-week intervals for a total of 12 visits each. Quarters infected with CNS were followed through time with milk samples being submitted for bacteriological culture and SCC determination. PCR amplification of the internal transcribed spacer region and sequencing of the sodA and rpoB genes were used for species allocation. Pulsed-field gel electrophoresis (PFGE) was performed to assess strain identity. The percentage of quarters affected per farm varied between 6 and 35%, with the most frequently isolated CNS species being Staphylococcus epidermidis, followed by Staph. simulans, Staph. chromogenes and Staph. haemolyticus. It was possible to follow 111 intramammary infections due to CNS through time. Duration of infection had a mean of 188 d and was not significantly different between CNS species. Geometric mean quarter SCC overall was 132 000 cells/ml and was also not significantly different between CNS species. Despite the possibility of a different epidemiology of infection, the impact in terms of udder health seems to be similar for different CNS species.

Nutritional, chemical and functional potential of Inga laurina (Fabaceae): A barely used edible species.

Inga laurina is a plant species which produces edible fruits, and until now there is little information available concerning its nutritional, chemical and bioactive composition. In this study, we evaluated for the first time the proximate composition and mineral contents in its fruit (peel, pulp and seed), that is the traditionally consumed part. The seeds obtained the highest protein (19.52 g/100 g), carbohydrate (22.5 g/100 g) and mineral contents, mainly Cu, Cr, P, Mn, Se and Zn. The peel and pulp were excellent sources of fiber (4.5 and 11.05 g/100 g) as well as mineral content, with Cr and Cu standing out in the pulp. This study is notably the first to provide a detailed assessment of the nutritional compositions of traditionally consumed and not consumed parts of this fruit. Sensory analysis of the pulp was also performed, which indicated good acceptance. The antioxidant properties were characterized in the fruit, peels and leaves. The ABTS test showed that leaf supernatant hydroethanolic crude extract (EC50 = 2.70 µg/mL) and its corresponding ethyl acetate (EC50 = 1.68 µg/mL) and butanol (EC50 = 2.48 µg/mL) partitions presented higher antioxidant potential compared to the control Ginkgo biloba (EC50 = 12.17 µg/mL). The most active precipitate extract regarding DPPH was from the peel (EC50 = 13.30 µg /mL) and the most active partition was the ethyl acetate (EC50 = 13.37 µg/mL), both with better activity compared to the control Ginkgo biloba (EC50 = 46.97 µg/mL). The ethyl acetate partition (EC50 = 13.45 µg/mL) and butanol partition (EC50 = 7.97 µg/mL) from the leaves showed the highest antioxidant capacity. Thus, extracts and partitions from the peels and leaves were studied from a phytochemical point of view due to presenting the best results for antioxidant capacity. The presence of phenolic compounds such as myricetin-3-O-rhamnopyranoside, myricetin-3-O-(2″-O-galloyl)-rhamnopyranoside and myricetin-3-O-(2″,4″-di-O-galloyl)-arabinopentoside-methyl ether were observed in the leaf crude extract and polar partitions, being reported for the first time in the Inga genus and Fabaceae family. Moreover, quercetin, quercetin-3-O-galatoctoside, quercetin-3-O-rhamnopyranoside, quercetin-3-O-(2″-O-galloyl)-rhamnoside, and quercetin tri-hexose were identified in the peel crude extract and ethyl acetate partition, in which the galloyl derivative of quercetin was identified for the first time in I. laurina fruit peels. GC-MS enabled separating and identifying substances such as palmitic and stearic acids, and ethyl oleate. It is possible to conclude that I. laurina pulp can be a supplementary food as a source of phenolic compounds, and the other organs of the plant (leaves and peel) are rich in flavonoids with great antioxidant capacity, making this species a promising source of antioxidants.

Zebrafish Avatar-test forecasts clinical response to chemotherapy in patients with colorectal cancer.

Cancer patients often undergo rounds of trial-and-error to find the most effective treatment because there is no test in the clinical practice for predicting therapy response. Here, we conduct a clinical study to validate the zebrafish patient-derived xenograft model (zAvatar) as a fast predictive platform for personalized treatment in colorectal cancer. zAvatars are generated with patient tumor cells, treated exactly with the same therapy as their corresponding patient and analyzed at single-cell resolution. By individually comparing the clinical responses of 55 patients with their zAvatar-test, we develop a decision tree model integrating tumor stage, zAvatar-apoptosis, and zAvatar-metastatic potential. This model accurately forecasts patient progression with 91% accuracy. Importantly, patients with a sensitive zAvatar-test exhibit longer progression-free survival compared to those with a resistant test. We propose the zAvatar-test as a rapid approach to guide clinical decisions, optimizing treatment options and improving the survival of cancer patients.

Food neophobia, risk perception and attitudes associations of Brazilian consumers towards non-conventional edible plants and research on sale promotional strategies.

Non-conventional edible plants (NCEP) are plants or parts of plants that are not usually consumed by the population and have limited geographic distribution. This study investigated the consumption of NCEP, the influence of Food Neophobia and risk perception on Brazilian consumers attitudes as well as purchase preferences to determinate the best sale promotional strategy. Participants (n = 271) answered the online questionnaire that consisted of socio-demographic questions, items about consumption (open questions), Food Neophobia Scale, risk perception (to assign the level risk) and attitudes towards NCEP (using 5-point Likert scale). Fisher's exact test was used to investigate possible associations. Task purchase choice was evaluated using the Best-Worst Scale. The most consumed conventional leafy vegetables were collard greens (Brassica oleracea L. var. acephala D.C) (95.6%) and lettuce (Lactuca sativa) (88.5%). As for NCEP, taioba (Xanthosoma taioba E.G) (26.7%), bertalha (Basela alba L.) (23.3%) and beldroega (Portulaca oleracea L.) (14.1%) were the most cited leafy vegetables. High food neophobia individuals demonstrated to try NCEP if its nutritional value and safety are proven, showing a demand for such strategy, this would increase the consumption of these vegetables. The perception of a high risk was also associated with the inclusion of NCEP in the diet. The inclusion of NCEP at the moment of purchase, even at a higher price, presented higher scores when compared to the purchase of only conventional vegetables, at lower prices. Despite this, the price can still be a limiting factor and the search for promotional strategies should be reinforced to increase the commercialization of NCEP at street markets.

Discriminant Analysis of Brazilian Stingless Bee Honey Reveals an Iron-Based Biogeographical Origin.

Stingless bee honey (SBH) is gaining attention due to its nutritional, sensorial, and medicinal characteristics. This study focuses on the combination of physicochemical properties, antioxidant capacity, mineral profile, and mass spectrometry-based fingerprints, using a chemometric approach to differentiate SBH (n = 18) from three different Brazilian biogeographical zones (Caatinga, Cerrado, and Atlantic Forest). The physicochemical properties of SBH varied, resulting in a wide range of water activity, moisture, total soluble solids, pH, and total and free acidity. The Caatinga honey showed the highest and the lowest contents of phenolics and flavonoids, respectively. The antioxidant free-radical scavenging assays demonstrated that the Brazilian SBH has a high antioxidant potential. The mineral profile of honey samples from the Atlantic Forest revealed higher contents of Ca and Fe while the Cerrado and Caatinga honey showed the highest P contents. Partial Least-Squares Discriminant Analysis (PLS-DA) analysis separated the samples into three groups based on the biogeographical zones of harvest. The main separation factors between groups were the m/z 326 ion and the Fe content. Univariate analysis confirmed that Fe content is important for SBH discrimination. The present results indicate that the origin of SBH can be determined on the basis of mineral profile, especially Fe content.

Behavioral and neuroinflammatory changes caused by glyphosate: Base herbicide in mice offspring.

BACKGROUND: Glyphosate is a pesticide considered of low toxicity, but scientific evidences show it can be harmful to health. This study aimed to evaluate the toxicity in mice offspring exposed to glyphosate-based herbicide (GBH) during the intrauterine period. METHODS: Female matrices received glyphosate 0.3 mg/kg daily per oral throughout the gestational period, which was variable between 18 and 22 days. From the 25th until the 28th days post-birth, mice offspring were subjected to behavioral tests, and the prefrontal cortex was processed for immunohistochemical analysis. RESULTS: Two significant behavioral changes were observed: anxiety in the GLIF0.3 group, increase in the behavior burying marbles in the marble-burying test and hyperactivity, expressed by the significant increase of the crossing number in the open field test. The increased microglia, TNF-alpha, and astrocyte expression were also observed in the prefrontal cortex of offspring treated with GLIF0.3. CONCLUSION: Exposure to GBH during mice intrauterine development induces hyperactive and anxious behavior, evidencing neuroinflammation.

Total Mercury Content in the Tissues of Freshwater Chelonium (Podocnemis expansa) and a Human Health Risk Assessment for the Amazon Population in Brazil.

Researchers recognize the silent, negative and deleterious effects caused by mercury pollution in gold mining areas. Freshwater turtles are culturally part of the diet of riverside populations in the Amazon region and this area presents mercury (Hg) pollution issues mainly due to gold mining activities. Thus, this research aimed to evaluate the total mercury (THg) content in the different organs of Amazonian giant river turtle (Podocnemis expansa) and carry out a human health risk assessment associated with the consumption of these animals. This study was conducted in the Vila Balbina, municipality of Presidente Figueiredo, state of Amazonas, Brazil. Skin (n = 28), muscle (n = 19) and brain (n = 2) samples were analyzed by Atomic Absorption Spectrometry (TDA-AAS) and a DMA-80™ mercury analyzer was used for the total mercury determinations. The average values found for THg in the skin, muscle and brain samples were, respectively, 0.1045 mg·kg-1, 0.1092 mg·kg-1 and 0.0601 mg·kg-1. Thus, THg was observed even though the P. expansa were kept in captivity, possibly due to previous contamination by air, water and food. The Hazard Quotient (HQ) was calculated considering a 9.07 g·day-1 intake dose of P. expansa and the consumption of turtles once a week showed an HQ = 2.45, which may cause long-term injuries to human health. Although the muscle concentrations were below the maximum limit established by the World Health Organization (WHO) and Brazilian regulatory agencies, it is important to evaluate consumption factors such as amount ingested, frequency and animal gender, which may cause a potential risk to regular consumers due to mercury bioaccumulation. The WHO may consider various aspects in order to warn the Amazon population about the severity and silent hazard of this metal, especially due to the importance of this matrix in the region. This region urgently needs government actions to inhibit clandestine mining and to prevent future serious, chronic health problems of the entire population.

Antimicrobial resistance and virulence profiles of Enterobacterales isolated from two-finger and three-finger sloths (Choloepus hoffmanni and Bradypus variegatus) of Costa Rica.

Background: Wildlife has been recently recognized as an environmental reservoir for antimicrobial resistance (AMR). However, less information on this topic is available in animals released back into the wild after rehabilitation in wildlife facilities, compared with studies performed exclusively in captive or free-ranging wildlife. This study aimed to evaluate the potential influence of captivity and/or treatment while in captivity of wild sloths on the AMR and virulence profiles of sloths' Enterobacterales. Methods: Oral and rectal swab samples were collected from 39 two-finger (Choloepus hoffmanni) and three-finger sloths (Bradypus variegatus) of Costa Rica (n = 78) and analyzed using conventional bacteriological techniques. A generalized linear mixed model was applied to estimate the isolates' multiple antimicrobial resistance and virulence indices as a function of animal status. Results: A considerable level of resistance was detected, especially for Citrobacter youngae and Escherichia coli, with 17.5% of isolates classified as multidrug-resistant. Virulence indices of isolates from rehabilitated sloths were significantly higher than the ones from sloths being hand-reared for shorter periods. Conclusions: To our knowledge, this is the first description of sloths' antimicrobial resistant Enterobacterales, suggesting that sloths' rehabilitation and consequent exposure to humans, may promote the selection of bacteria with higher virulence. Ultimately, these bacteria may represent a threat to human and animal health due to their zoonotic potential and AMR and virulence profiles.

Spillover events of rabbit haemorrhagic disease virus 2 (recombinant GI.4P-GI.2) from Lagomorpha to Eurasian badger.

Rabbit haemorrhagic disease (RHD) is a major threat to domestic and wild European rabbits. Presently, in Europe, the disease is caused mainly by Rabbit haemorrhagic disease virus 2 (RHDV2/b or Lagovirus europaeus GI.2), the origin of which is still unclear, as no RHDV2 reservoir hosts were identified. After the RHDV2 emergence in 2010, viral RNA was detected in a few rodent species. Furthermore, RHDV2 was found to cause disease in some hare species resembling the disease in rabbits, evidencing the ability of the virus to cross the species barrier. In this study, through molecular, histopathologic, antigenic and morphological evidences, we demonstrate the presence and replication of RHDV2 in Eurasian badgers (Meles meles) found dead in the district of Santarém, Portugal, between March 2017 and January 2020. In these animals, we further classify the RHDV2 as a Lagovirus europaeus recombinant GI.4P-GI.2. Our results indicate that Meles meles is susceptible to RHDV2, developing systemic infection, and excreting the virus in the faeces. Given the high viral loads seen in several organs and matrices, we believe that transmission to the wild rabbit is likely. Furthermore, transmission electron microscopy data show the presence of calicivirus compatible virions in the nucleus of hepatocytes, which constitutes a paradigm shift for caliciviruses' replication cycle.

Study of inorganic elements in different organs and tissues of Amazonian manatee (Trichechus inunguis) from Brazil.

Multielement concentrations (P, S, Cl, K, Ca, Cr, Mn, Fe, Ni, Cu, Zn, Rb, and Rh) and total mercury (T-Hg) were analyzed in different organs and tissues of Amazonian manatee (Trichechus inunguis). Samples of 27 T. inunguis specimens, maintained in the collection of the Amazonian Center for the Research and Preservation of Aquatic Mammals, were used, situated in an area highly impacted by gold mining in the northern region of the Brazilian Amazon. Samples of aquatic plants used as food by the animals were also analyzed. The elements S, Cl, K, Cr, and Mn accumulated mainly in the musculature, while Fe and Cu were more concentrated in the liver. Trace elements, such as rubidium (Rb) and rhodium (Rh), not previously reported in the organs of animals of the family Trichechidae, were also identified. The averages for T-Hg in the skin, muscle, encephalon, liver, kidney, and lung samples were, respectively, 0.1540 ± 0.1332, 0.0593 ± 0.1044, 0.0517 ± 0.0467, 0.0486 ± 0.0543, 0.0237 ± 0.0336, and 0.0013 ± 0.0032 µg.g-1. The values obtained for the vibrissae samples were below the limit of quantification, which allows for the conclusion that this tissue cannot be used as a contamination marker. It was observed that even when kept in a conservation breeding site, these animals were exposed to non-essential trace elements. Differences in the accumulation of elements were observed between the different organs and tissues analyzed. The presence of contaminants in animals that live in a preservation center, even at low levels, deserves attention.