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Applications and advances in surface-enhanced Raman spectroscopy (SERS) have grown over the last several years, demonstrating improvements in sensitivity and selectivity. Applications for SERS-related techniques have been explored in many analytical disciplines including forensic applications. Drug chemistry and toxicology may benefit from the growth of SERS since many methods have demonstrated high sensitivity, potential for quantitative analysis, and portable instrumentation for onsite testing. SERS techniques have been developed using many substrates including nanoparticles and colloids; microfluidic devices; paper-based substrates; electrodes; and wearable/flexible devices. These methods take advantage of the SERS phenomenon, providing more potential applications for Raman analysis and overcoming the traditional challenges of the technique. This is desirable for forensic drug chemistry applications which require screening and confirmatory approaches and analysis of mixtures with low weight percent contributions of analytes, as well as forensic toxicology, where matrix interferences, sensitivity requirements, and a growing introduction of novel drug entities pose challenges. Several chemometric approaches have been applied to Raman data to improve identification, classification, and quantification. These advances position SERS for incorporation into forensic chemistry laboratories. While some SERS applications are available commercially, widespread use of SERS in practicing forensic laboratories remains limited. Due to the requirements for use of analytical techniques in the courtroom, future studies for SERS should revolve around the assessment of validation parameters common in other analytical methods. SERS techniques that are rapid, simple, and inexpensive may be more quickly adapted for forensic testing, where use as advanced screening techniques is a clear avenue of research. This article is categorized under: Forensic Chemistry and Trace Evidence > Controlled and Emerging Drug Compounds Toxicology > New Psychoactive Substances Forensic Chemistry and Trace Evidence > Emerging Technologies and Methods. [ABSTRACT FROM AUTHOR]

Background: The association of microbiota with clinical outcomes and the taxa associated with colitis in horses remains generally unknown. Objectives: Describe the fecal microbiota of horses with colitis and investigate the association of the fecal microbiota with the development of laminitis and survival. Animals: Thirty‐six healthy and 55 colitis horses subdivided into laminitis (n = 15) and non‐laminitis (n = 39, 1 horse with chronic laminitis was removed from this comparison) and survivors (n = 27) and nonsurvivors (n = 28). Methods: Unmatched case‐control study. The Illumina MiSeq platform targeting the V4 region of the 16S ribosomal RNA gene was used to assess the microbiota. Results: The community membership (Jaccard index) and structure (Yue and Clayton index) were different (analysis of molecular variance [AMOVA]; P <.001) between healthy and colitis horses. The linear discriminant analysis effect size (LEfSe; linear discriminant analysis [LDA] >3; P <.05) and random forest analyses found Enterobacteriaceae, Lactobacillus, Streptococcus, and Enterococcus enriched in colitis horses, whereas Treponema, Faecalibacterium, Ruminococcaceae, and Lachnospiraceae were enriched in healthy horses. The community membership and structure of colitis horses with or without laminitis was (AMOVA; P >.05). Enterobacteriaceae, Streptococcus, and Lactobacillus were enriched in horses with laminitis (LDA > 3; P <.05). The community membership (AMOVA; P =.008) of surviving and nonsurviving horses was different. Nonsurviving horses had an enrichment of Enterobacteriaceae, Pseudomonas, Streptococcus, and Enterococcus (LDA >3; P <.05). Conclusion and Clinical Importance: Differences in the microbiota of horses with colitis that survive or do not survive are minor and, similarly, the microbiota differences in horses with colitis that do or do not develop laminitis are minor. [ABSTRACT FROM AUTHOR]

Our paper intends to recognize the presence of the thopic civilizationsavagism in two books dealing with the mapuche people from the Araucanía region in Chili. Before reaching that goal we are constructing a theoretical approach to its two nouns as well as making references to context issues. The effect of the thopic in history became essentially rewarding for some people, the europeans, but evidently devastating for others, specially the native americans and the africans. The thopic's verbal components are extremely simple: Civilization Noun + Savagism Noun = Topic C -S. Soon became a powerful enunciation that imposed itself by the action of its followers in large are as upon the earth during the XIX and XX centuries. The books where we show its presence are the novels Flor Lumao and El vado de la noche by Lautaro Yankas (1932 and 1954 respectively). [ABSTRACT FROM AUTHOR]

This study developed a reliable laser‐induced breakdown spectroscopy (LIBS) screening approach capable of detecting GSR in just a few minutes with minimal damage to the sample, high specificity, and sensitivity. Moreover, a novel micro‐sampling method was developed to gather three‐dimensional data of the simultaneous occurrence of IGSR markers from a discrete space. The method is capable of micro‐spatial chemical analysis from just two laser shots fired at an area of 100‐μm diameter. The performance of the micro‐spot method is compared with our previously published bulk‐line method. Superior accuracy, spatial information of IGSR distribution in the sample, and a less invasive sampling are some of the advantages of the newly proposed method. A benefit afforded by this approach is the use of the universal hand's collection method currently used by practitioners, while leaving over 99% of the stub left unaltered for further analysis. Machine learning algorithms were used for the classification of samples derived from shooters' hands versus nonshooters hands, based on their LIBS spectrochemical data. Four different approaches—critical threshold, logistic regression, naïve Bayes, and neural networks—were applied to examine the performance and accuracy of two different ablation patterns (micro‐spot and bulk‐line mode). A validation set of 326 samples originated from 51 nonshooters and 56 known shooters resulted in an overall accuracy between 87% and 100%, depending on the ablation pattern and the type of prediction model applied. The incorporation of this rapid screening and statistical decision‐making approach could offer more efficient case management in firearm‐related investigations. A LIBS method capable of documenting the distribution of IGSR markers on substrates dabbed from an individual's hands was developed. The 3D data include simultaneous information about the analyte characteristic wavelengths, their respective intensity, and x‐y location coordinates within the sample. Logistic regression, naïve Bayes, and neural network machine learning models provided effective predictive methods, with classification accuracy better than 93.7% for a validation set of 326 authentic samples collected from shooters and background populations. [ABSTRACT FROM AUTHOR]

Abstract This study was conducted to assess the catalytic electrode surface adsorption and capture properties of different metal chitosan derivatives in aqueous phosphate buffer solution (pH = 7.3). Early, recent work showed that the response of Iron chitosan complex with R = -CH 3 on the periphery, over blood red cells in presence of sphingomyelinase C was protected. The effect of others substituent (R = -Br, -Cl, -F, NO 2 , -OCH 3 , -H) on the periphery of the Schiff base ligand did not show correlation with the oxidation of sphingomyelinase C and its biological response. For this reason, various adsorbed metal (M = Fe of recent work, Cu, Ni and Co) complexes of chitosan and Schiff bases on glassy carbon electrode for the oxidation of sphingomyelinase C were investigated and compared, each one with -CH 3 group on the periphery of the Schiff base. UV–Vis and IR-TF spectroscopies, electrochemistry and microscopy assay were performed; then, the metal effect underlying. For the Schiff base, cobalt and copper complexes did not proved to be a remarkable cellular protector in presence of the enzyme, but the nickel complex showed to be a cellular protector at short time, this conclusion help to proposal a reaction mechanism for the electrochemical and biological studies. Highlights • The inhibition of Sphingomyelinase C by metal chitosan complexes was examined. • Metal chitosan complexes were active for the oxidation of SMASE. • NiChi-CH 3 protected to erythrocytes exposed to Sphingomyelinase C at short times. • Reactions for SMASE oxidation by MChi-CH 3 were proposed. [ABSTRACT FROM AUTHOR]

This study evaluated the impact of an algorithm targeting antimicrobial therapy of diarrhoeic calves on the incidence of diarrhoea, antimicrobial treatment rates, overall mortality, mortality of diarrhoeic calves and changes in the faecal microbiota. The algorithm was designed to target antimicrobial therapy in systemically ill calves from on two dairy farms. Retrospective (farm 1: 529 calves; farm 2: 639 calves) and prospective (farm 1: 639 calves; farm 2: 842 calves) cohorts were examined for 12 months before and after implementation of the algorithm. The Mantel–Haenszel test and Kaplan–Meier survival curves were used to assess the cumulative incidence risk (CIR) and time to development of each outcome before and after implementation of the algorithm. The CIR of antimicrobial treatment rates was 80% lower after implementation of the algorithm on both farms (CIR 0.19, 95% confidence interval 0.17–0.21). There was no difference in the CIR of overall mortality, but the CRI for mortality of diarrhoeic calves was lower in the period after implementation of the algorithm on one farm. The faecal microbiota of 15 healthy calves from both farms at each time period were characterised using a sequencing platform targeting the V4 region of the 16S rRNA gene. On both farms, there were significant differences in community membership and structure (parsimony P < 0.001). Use of the algorithm for treatment of diarrhoeic calves reduced antimicrobial treatment rates without a negative impact on the health of calves. However, the experimental design did not take into account the potential confounding effects of dietary changes between the study periods. [ABSTRACT FROM AUTHOR]

Identification of genotypes with acceptable yield and yield stability in different environments is an important issue in plant breeding. Genotype-by-environment interaction (GEI) can alter genotypes performances making the selection of superior material a tedious task for breeders. Consequently, it is necessary to assess the usefulness of different available methods and identify the most suitable for understanding GEI. The objectives of this work were to compare three methods to study genotype stability considering incomplete data sets: (i) Di Rienzo, Guzmán and Casanoves’ test (DGC), (ii) relative yield (RY) and (iii) Piepho’s method. In addition, AMMI (additive main effect and multiplicative interaction) analysis and eight AMMI stability measures SIPC, EV, ASV, Da, FP, B, FA and Za were computed to explore their advantages and disadvantages to select stable entries. The usefulness of the genotype selection index (GSI) and the rank-sum (RS) procedures to identify stable and high-yielding genotypes were evaluated and then compared with the superiority (P) and reliability indexes (I).The association between yield variation and climatic factors as frosts, chilling, heat, rainfall and the interactions among them were also analyzed. 29 peach entries were assessed in four to seven seasons in a completely randomized design with three replications. DGC and RY tests agreed on classifying Fireprince as a stable and high-yielding peach, RY classified 25 entries as stable, while Piepho’s method did not separate the tested genotypes as DGC and RY did. The results of AMMI indicated that 25.06% of total variability was justified by genotypes, 9.76% by environments and 58.97% by GEI. The first five interaction principal components could explain 94.82% of GEI and showed the efficiency of AMMI model to study and understand GEI. The AMMI parameters showed no association with fruit yield, therefore, they could be useful to indicate stable entries but they would not be appropriate to select stable and high-yielding genotypes. The EV and Za indicated static stability while ASV, SIPC, Da, FA and FP pointed out the dynamic stability concept. The performance of the best entries selected by GSI, RS, P and I procedures were not different, therefore, any of them can be used to select superior peach genotypes. Rainfall during endodormancy, rainfall from floral bud endo- to ecodormancy - and heat accumulation during fruit development period showed significant correlation with yield variation across seasons. [ABSTRACT FROM AUTHOR]

Nowadays, although there are several studies based on peach phenotypic diversity and adaptability, there is no current research regarding associations among characters using multivariate statistical methods which provide more trustable conclusions than bivariate analysis. For this reason, we evaluated not only the adaptability of the accessions but also the relationships among qualitative and quantitative traits applying Multiple Correspondence Analysis and Factor Analysis, respectively. The study was carried out on seventy-seven nectarine and one hundred and thirty-two peach accessions grown in Argentina that were characterized with 35 fruit quality, agronomic and phenological traits during 3 seasons. Multiple Correspondence Analysis revealed that freestone trait would be associated with strong anthocyanin coloration of the flesh around the brown dark stone while semi-freestone trait would be related to weak or no anthocyanin coloration of the flesh around the light brown stone. Then, Factor Analysis enabled us to identify positive relationships between blooming and sprouting date; harvesting period and yield and among fruit development period, harvesting date, fruit weight and soluble solid content. In addition, ‘phenology traits’, ‘productivity’ and ‘fruit quality and reaping time’ would be unrelated to each other. Furthermore, the agglomerative hierarchical cluster analysis grouped the accessions in accordance to their similarities and allowed the identification of the most adapted cultivars. We have demonstrated that this approach is a powerful tool that should be considered for further studies on peach germplasm and it has also the possibility to be applied for other fruit species. Finally, the results of this study provide valuable information for germplasm characterization and for breeding practices aiming to develop accessions adapted to the climatic conditions of the northern of Buenos Aires province. [ABSTRACT FROM AUTHOR]

Theiler's disease (TD) is a (sub-)acute hepatitis in adult horses and one of the most common causes of acute hepatic failure. Recent findings indicate that equine parvovirus hepatitis (EqPV-H) likely causes TD and that its transmission occurs via iatrogenic and/or natural routes. Following the death of an EqPV-H positive mare with TD, close-contact mares and foals in the same paddock were monitored to evaluate if there was any evidence of EqPV-H. For this purpose, the serum of close contact horses was examined 6 and 42 days after the mare's death for the presence of EqPV-H DNA and changes in liver-associated serum biochemical parameters. The foals had higher EqPV-H viral loads than the mares. Apart from the mare that was euthanized, none of the horses included in this study showed signs of severe disease and nor did they have particularly elevated liver enzymes. Nucleotide sequence analysis revealed no major differences between the viral DNA detected in the serum of the dead mare and any of the in-contact horses. In conclusion, our data confirmed previous findings that horizontal transmission of EqPV-H may occur through close contact between horses. • Horizontal transmission of EqPV-H to close contact horses. • Foals display higher viral loads compared to mares. • None of the horses showed signs of severe disease with elevated liver enzymes. [ABSTRACT FROM AUTHOR]

Clostridioides difficile (formerly Clostridium difficile) is associated with colitis in foals and mature horses. C. difficile exposes specific phosphorylated polysaccharides (PSs), named PS-I, PS-II and PS-III. These cell-surface PSs are potential vaccine targets, especially the hexasaccharide phosphate PS-II, that has been found in all C. difficile ribotypes examined. Since we previously identified anti-PS-II circulating antibodies in horses, we postulated that vaccinating foals with PS-II may prevent colonization by C. difficile. In this study, we aim to evaluate the IgM antibody responses in foals to PS-II. To evaluate the reactogenicity and immunogenicity of C. difficile PS-II in foals, three-to four-month-old foals were vaccinated intramuscularly three times at intervals of three weeks with 100 μg/dose (3 foals) or 500 μg/dose (3 foals) of purified PS-II antigen with aluminum hydroxide adjuvant, or with a placebo preparation (2 foals) containing adjuvant alone. No injection site swelling, pain or fever was observed after vaccination. Two of the three foals receiving 100 μg/dose, and three out of three foals receiving 500 μg/dose of PS-II responded with increases in serum IgM antibodies. No control foals that received the placebo had IgM responses to PS-II. There was a trend towards a higher response rate in foals receiving 500 μg PS-II one week after second vaccination when compared to control foals and towards higher concentrations of serum IgM antibodies in foals receiving 500 μg PS-II. No adverse reactions were observed following vaccination with PS-II in foals; Serum IgM immune responses were induced by vaccination. A polysaccharide-based vaccine for C. difficile in horses deserves further investigation. • Clostridioides difficile polysaccharide PS-II is immunogenic in foals. • Vaccination of foals with C. difficile PS-II raises anti-PS-II serum IgM antibodies. • No adverse reactions were observed following vaccination with PS-II in foals. [ABSTRACT FROM AUTHOR]

Multiple analytical techniques for the screening of fentanyl‐related compounds exist. High discriminatory methods such as GC–MS and LC–MS are expensive, time‐consuming, and less amenable to onsite analysis. Raman spectroscopy provides a rapid, inexpensive alternative. Raman variants such as electrochemical surface‐enhanced Raman scattering (EC‐SERS) can provide signal enhancements with 1010 magnitudes, allowing for the detection of low‐concentration analytes, otherwise undetected using conventional Raman. Library search algorithms embedded in instruments utilizing SERS may suffer from accuracy when multicomponent mixtures involving fentanyl derivatives are analyzed. The complexing of machine learning techniques to Raman spectra demonstrates an improvement in the discrimination of drugs even when present in multicomponent mixtures of various ratios. Additionally, these algorithms are capable of identifying spectral features difficult to detect by manual comparisons. Therefore, the goal of this study was to evaluate fentanyl‐related compounds and other drugs of abuse using EC‐SERS and to process the acquired data using machine learning—convolutional neural networks (CNN). The CNN was created using Keras v 2.4.0 with Tensorflow v 2.9.1 backend. In‐house binary mixtures and authentic adjudicated case samples were used to evaluate the created machine‐learning models. The overall accuracy of the model was 98.4 ± 0.1% after 10‐fold cross‐validation. The correct identification for the in‐house binary mixtures was 92%, while the authentic case samples were 85%. The high accuracies achieved in this study demonstrate the advantage of using machine learning to process spectral data when screening seized drug materials comprised of multiple components. [ABSTRACT FROM AUTHOR]

Background: The composition of the microbiota of the equine intestinal tract is complex. Determining whether the microbial composition of fecal samples is representative of proximal compartments of the digestive tract could greatly simplify future studies. The objectives of this study were to compare the microbial populations of the duodenum, ileum, cecum, colon and rectum (feces) within and between healthy horses, and to determine whether rectal (fecal) samples are representative of proximal segments of the gastrointestinal tract. Intestinal samples were collected from ten euthanized horses. 16S rRNA gene PCR-based TRFLP was used to investigate microbiota richness in various segments of the gastrointestinal tract, and dice similarity indices were calculated to compare the samples. Results: Within horses large variations of microbial populations along the gastrointestinal tract were seen. The microbiota in rectal samples was only partially representative of other intestinal compartments. The highest similarity was obtained when feces were compared to the cecum. Large compartmental variations were also seen when microbial populations were compared between six horses with similar dietary and housing management. Conclusion: Rectal samples were not entirely representative of intestinal compartments in the small or large intestine. This should be taken into account when designing studies using fecal sampling to assess other intestinal compartments. Similarity between horses with similar dietary and husbandry management was also limited, suggesting that parts of the intestinal microbiota were unique to each animal in this study. [ABSTRACT FROM AUTHOR]

The intestinal tract houses one of the richest and most complex microbial populations on the planet, and plays a critical role in health and a wide range of diseases. Limited studies using new sequencing technologies in horses are available. The objective of this study was to characterize the fecal microbiome of healthy horses and to compare the fecal microbiome of healthy horses to that of horses with undifferentiated colitis. A total of 195,748 sequences obtained from 6 healthy horses and 10 horses affected by undifferentiated colitis were analyzed. Firmicutes predominated (68%) among healthy horses followed by Bacteroidetes (14%) and Proteobacteria (10%). In contrast, Bacteroidetes (40%) was the most abundant phylum among horses with colitis, followed by Firmicutes (30%) and Proteobacteria (18%). Healthy horses had a significantly higher relative abundance of Actinobacteria and Spirochaetes while horses with colitis had significantly more Fusobacteria. Members of the Clostridia class were more abundant in healthy horses. Members of the Lachnospiraceae family were the most frequently shared among healthy individuals. The species richness reported here indicates the complexity of the equine intestinal microbiome. The predominance of Clostridia demonstrates the importance of this group of bacteria in healthy horses. The marked differences in the microbiome between healthy horses and horses with colitis indicate that colitis may be a disease of gut dysbiosis, rather than one that occurs simply through overgrowth of an individual pathogen. [ABSTRACT FROM AUTHOR]

Background: Clostridium difficile and Clostridium perfringens are commonly associated with colitis in equids, but healthy carriers exist. Scarce information is available on the prevalence of Clostridium spp. in gastrointestinal compartments other than faeces in healthy horses, and it is unknown whether faecal samples are representative of proximal compartments. The objectives were to investigate the prevalence of C. difficile and C. perfringens in different intestinal compartments of healthy adult horses and to determine whether faecal samples are representative of colonization in proximal sites and overall carrier status. Results: Toxigenic C. difficile was isolated from 14/135 (10.3%) samples from 8/15 (53.3%) horses. Between zero and three sites were positive per horse, and multiple sites were positive in four horses. Isolates were recovered from duodenum, jejunum, ileum, right dorsal colon, small colon and rectum. When multiple compartments were positive in a single horse, two different C. difficile ribotypes were always present. Clostridium perfringens Type A (CPE, β2 toxin gene negative) was recovered from the left ventral colon of one horse (0.74%, 1/135 samples). Agreement between faeces and overall C. difficile carrier status was good. Conclusions: Clostridium difficile can be found in different compartments of the gastrointestinal tract of healthy horses, and multiple strains can be present in an individual horse. The prevalence of C. perfringens in healthy adult hoses was low, consistent with previous reports. Faecal samples were representative for presence of C. difficile in proximal compartments in 5/8 horses (63%) but were not representative for the specific strain. [ABSTRACT FROM AUTHOR]

This article describes the applicability of a rapid laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) method for the analysis of soil and sediment samples with broad chemical and physical properties and the comparison of its analytical performance to digestion protocols commonly used in environmental sciences. Two sets of samples were evaluated, contaminated soils from a residential area and undisturbed sediments from natural environments. Analytical results obtained by LA-ICP-MS correlate well at the 95% confidence level to total digestion followed by inductively coupled plasma optical emission spectroscopy (ICP-OES) and ICP-MS analysis. A total of 48 sediments collected in South Florida were also analyzed for screening purposes and to evaluate the application of LA-ICP-MS in environmental forensics. Normalization using Al combined with non-parametric correlation tests and principal component analyses were successfully used to predict correlations between data acquired by LA-ICP-MS and by partial digestion methods followed by ICP methods. Precision and accuracy for the LA method was <20%, which is typically accepted for digestion methods of soils and sediments. The overall bias, evaluated on reference standard materials, ranged from 8%-15%. The overall precision obtained on samples was <10% relative standard deviation (RSD). [ABSTRACT FROM AUTHOR]

Abstract: A simple, rapid and sensitive method was developed for the routine analysis of trace elements on sediments and soils by UV-ns-LA-ICP-MS. The homogenization procedure that reduces the particle size of the samples to less than 1 µm diameter was found to be a key factor to allow for a representative sampling of the bulk soil at the micro-scale and to improve reproducibility and cohesion of the sample without requiring the use of any binder. The elimination of binders simplified the sample preparation and avoided any undesirable dilution of the sample. SEM/EDX analyses were conducted to evaluate the efficiency of laser to sample interaction. Matrix effects and different quantitation strategies were employed to demonstrate the utility of the analytical technique. Sixteen elements were analyzed on soil and sediment samples and certified reference materials. Analytical results obtained by LA-ICP-MS were comparable to solution ICP-MS analysis in terms of accuracy, precision and limits of detection. Two independent proficiency tests for trace metals in soils were conducted to compare the performance of the method versus conventional digestion ICP and AA methods, obtaining z scores ≤3 for all elements measured by LA-ICP-MS. An overall bias between 8 and 15% was found, depending on the sample while the overall precision was found to be better than 5% RSD for all samples. Limits of detection were as low as 0.01 mg kg−1. [Copyright &y& Elsevier]

The chemical and biological properties of compost made from yard trimmings (YT) composted alone or mixed with slaughterhouse wastes (SHW) were evaluated in seven phases. Mixtures were weighed in a 2:1 proportion (YT:SHW) and placed in composting bins (0.91m2). Temperature was recorded to determine the time (d) needed to reach the first (1HC) and second heat cycles (2HC). Composting characteristics were measured at 0 d, at the peak of the 1HC and 2HC, and at maturation (0, 20, 50 and 70 d). During 1HC, bacterial isolates were cultivated in both treatments and identified using the Biolog® System. Chemical composition was statistically analyzed using a 2 (layers of SHW)× 7 (composting phases) factorial arrangement of treatments with the ANOVA procedure of SAS. The pH was neutral for YT and ranged from 7.41 to 6.82 for SHW throughout the process. There was a decrease in organic matter (OM) and carbon (C), and a relative increase in nitrogen (N) in both treatments. At 70 d of maturation, C:N values were similar between treatments, but lower (P >0.05) than the initial values. Final N concentration was higher (P >0.05) for the treatment with SHW. Only the SHW treatment exhibited thermophilic temperatures. At the 1HC in both treatments, different populations of bacteria responsible for the breakdown of OM were identified showing an active heterogeneous population. The presence of pathogenic microorganisms was not detected in treatments containing SHW. [Copyright &y& Elsevier]

Abstract: Clostridium difficile is an important cause of diarrhea in horses, causing sporadic and epidemic disease of varying severity. This study evaluated the molecular characteristics of 48 C. difficile isolates recovered from diarrheic horses admitted to a veterinary hospital by using PCR-ribotyping and toxin gene profile. Additionally, feces were tested for the presence of C. difficile toxin A/B via enzyme immunosorbant assay (EIA) in 38 horses. The toxin genes tcdA, tcdB and cdtB were present in 27 (56.25%), 35 (72.91%) and 2 (4.1%) strains, respectively. Eight isolates (16.6%) were A−B+ variants. Thirteen of forty-eight isolates (27.0%) did not posses any toxin genes (A−B−CDT−). A positive EIA result was reported in 17 (44%) of the cases. There was no association between the presence of different ribotypes or strains and toxin gene(s) profiles and the clinical outcome. [Copyright &y& Elsevier]

Case Description—4 racehorses were examined because of markedly abnormal behavior following administration of fluphenazine decanoate. Clinical Findings—Clinical signs included restlessness, agitation, profuse sweating, hypermetria, aimless circling, intense pawing and striking with the thoracic limbs, and rhythmic swinging of the head and neck alternating with episodes of severe stupor. Fluphenazine was detected in serum or plasma from all 4 horses. The dose of fluphenazine decanoate administered to 3 of the 4 horses was within the range (25 to 50 mg) routinely administered to adult humans. Treatment and Outcome—In 2 horses, there was no response to IV administration of diphenhydramine hydrochloride, but the abnormal behavior in these 2 horses appeared to resolve following administration of benztropine mesylate, and both horses returned to racing. The other 2 horses responded to diphenhydramine administration. One returned to racing. The other was euthanized because of severe neurologic signs, respiratory failure, and acute renal failure. Clinical Relevance—Findings indicate that adverse extrapyramidal effects may occur in horses given fluphenazine decanoate. These effects appear to be unpredictable and may be severe and life threatening. Use of fluphenazine decanoate as an anxiolytic in performance horses is not permitted in many racing and horse show jurisdictions, and analytic procedures are now available to detect the presence of fluphenazine in serum or plasma. [ABSTRACT FROM AUTHOR]

Simple Summary: DNA sequencing has been used to characterize the intestinal microbiota of horses. However, all published studies are based on Illumina technology to date because it is widely accessible, but a major limitation is the short bacterial DNA fragment sequenced. Other long-read sequencing technologies (i.e., PacBio) can classify bacteria at the species level and therefore are more meaningful for the identification of specific changes in the gut after the use of antimicrobials, for example. The objectives of this study were to characterize the gut microbiota of horses at the species level before and after trimethoprim sulfadiazine administration and to compare results with short-read Illumina sequencing. PacBio sequencing failed to classify the equine intestinal microbiota at the species level but detected greater richness and less unclassified bacteria compared to Illumina sequencing. Bacteroidetes were the most abundant bacteria in the feces, followed by Firmicutes and Fibrobacteres in this study. Further studies should focus on improving databanks for equine studies. All current studies have used Illumina short-read sequencing to characterize the equine intestinal microbiota. Long-read sequencing can classify bacteria at the species level. The objectives of this study were to characterize the gut microbiota of horses at the species level before and after trimethoprim sulfadiazine (TMS) administration and to compare results with Illumina sequencing. Nine horses received TMS (30 mg/kg) orally for 5 days twice a day to induce dysbiosis. Illumina sequencing of the V4 region or full-length PacBio sequencing of the 16S rRNA gene was performed in fecal samples collected before and after antibiotic administration. The relative abundance and alpha diversity were compared between the two technologies. PacBio failed to classify the equine intestinal microbiota at the species level but confirmed Bacteroidetes as the most abundant bacteria in the feces of the studied horses, followed by Firmicutes and Fibrobacteres. An unknown species of the Bacteroidales order was highly abundant (13%) and deserves further investigation. In conclusion, PacBio was not suitable to classify the equine microbiota species but detected greater richness and less unclassified bacteria. Further efforts in improving current databanks to be used in equine studies are necessary. [ABSTRACT FROM AUTHOR]

Background: This study aimed to describe and compare therapeutic approaches for horses with acute diarrhea in different geographic regions worldwide. Methods: Clinical information was retrospectively collected from diarrheic horses presented to participating institutions between 2016 and 2020, including fluid therapy on admission, antimicrobial drugs, probiotics, anti-endotoxic medications, anti-inflammatory drugs, gastroprotectants, digital cryotherapy, and toxin-binding agents. Seasonal and geographic differences were investigated. Results: 1438 horses from 26 participating hospitals from 5 continents were included. On admission, 65% (926/1419) of horses were administered a fluid bolus. Antimicrobial drugs were administered to 55% (792/1419) within the first 24 hours of admission, with penicillin and gentamicin being the most used combination (25%, 198/792). Horses with leukopenia (OR: 2.264, 95%CI: 1.754 to 2.921; P<0.001) or meeting systemic inflammatory response syndrome criteria (OR: 2.542, 95%CI: 1.919 to 3.368; P<0.001) had higher odds of being administered antimicrobial drugs. Other treatments administered included probiotics (15%, 215/1438), polymyxin B (13%; 187/1438), pentoxifylline (8%; 118/1438), gastroprotectants (44%; 626/1419), digital cryotherapy (34%; 489/1435), plasma transfusion (13%; 182/1410) and toxin-binding agents (36%; 515/1438). Limitations: Due to the retrospective nature of the study, the rationale for treatment decisions was unavailable, and associations with survival could not be evaluated. Conclusions: Treatments varied between hospitals from different geographic regions. Prospective clinical trials are required to evaluate the effects of various treatments on survival. [ABSTRACT FROM AUTHOR]

The identification of counterfeit pharmaceutical packaging is a complex problem that often requires multiple physical or chemical analyses. In this study, LIBS and ATR-FTIR were used as fast and non-invasive methods to classify pharmaceutical paperboard packaging sample into an authentic group or belonging to one of five counterfeit counterpart packaging sources. The collection set consisted of 124 external pharmaceutical packages, originating from 6 authentic sources (n = 12 packages) and 3 to 5 counterfeit sources per product (n = 112 packages). Orthogonal methods, such as LIBS and ATR-FTIR, were especially advantageous for classification purposes because they reveal chemical information of the organic and inorganic components of the package ink. Different ink colors from the logo, text, barcodes, and images were tested; additionally, the paperboard sample substrate was analyzed with ATR-FTIR. After data reduction via Principal Component Analysis, two supervised machine learning classification techniques were used for classification: k -Nearest Neighbors, and Linear Discriminant Analysis. A random 60:40 split of the data was used for training and testing the algorithms. Across all analyzed ink types, correct classification rates above 70% (LIBS) and 85% (ATR-FTIR) were observed. Data fusion of the two complementary methods improved the results, providing correct classifications ranging from 90 to 100% depending on the classifier and the pharmaceutical package. The results demonstrate that the elemental information obtained by LIBS and the chemical composition by ATR-FTIR are complementary and provide practical alternatives for fast screening of counterfeit secondary packaging. Unlabelled Image • LIBS and ATR-FTIR spectral profiles differentiated authentic versus counterfeit ink packaging. • Data fusion of LIBS and IR yielded more accurate classifications (>90%) • LIBS and ATR-FTIR are quick approaches for direct and non-invasive screening of counterfeit packages. [ABSTRACT FROM AUTHOR]

Simple Summary: Acute diarrhea (colitis) is a major problem in adult horses and the role of the intestinal bacteria (microbiota) is still poorly understood in this species. The aim of this study was to compare the mucosal and luminal content microbiota of the cecum and colon of healthy and diarrheic horses. We concluded that microbial dysbiosis (changes in the normal microbiota composition) occurs in horses with colitis at different levels of the intestinal tract and microbiota composition is different between the mucosa and luminal content of diarrheic horses. Changes in species associated with dysbiosis could be used in the future for disease diagnosis, prognosis and treatment of equine colitis. The aim of this study was to compare the mucosal and luminal content microbiota of the cecum and colon of healthy and diarrheic horses. Marked differences in the richness and in the community composition between the mucosal and luminal microbiota of the cecum and large colon of horses with colitis were observed. Microbial dysbiosis occurs in horses with colitis at different levels of the intestinal tract, and microbiota composition is different between the mucosa and luminal content of diarrheic horses. The changes in some key taxa associated with dysbiosis in the equine intestinal microbiota, such as Escherichia, Fusobacterium and Lactobacillus, deserve further inquiry in order to determine their utility for disease diagnosis and treatment. [ABSTRACT FROM AUTHOR]

Background: Increased concentration of haptoglobin (Hp) in serum is associated with survival of critically ill humans and horses. High serum activity of liver‐derived enzyme is associated with sepsis in children and foals. Hypothesis/Objectives: Investigate whether admission serum Hp and glutamic dehydrogenase (GLDH) are associated with systemic inflammatory response syndrome (SIRS) and survival of sick calves. Animals: One hundred two calves. Methods: Retrospective cross‐sectional study. Electronic medical records from all calves <30 days of age admitted to a teaching hospital for 8 years were reviewed. The signalment, clinicopathological findings, the presence of SIRS, final diagnosis, hospitalization time and outcome were recorded. A Cox proportional hazard ratio (HzR) were calculated to assess the association between clinicopathological variables and survival to discharge. Results: Serum Hp concentrations were similar between SIRS (0.29 g/L; range, 0.05‐3.6) and non‐SIRS calves (0.22 g/L; range, 0‐4.2; P =.62). GLDH activity was similar between SIRS (12 U/L; range, 1‐1025) and non‐SIRS calves (9 U/L; range, 2‐137; P =.2). Absent suckle reflex (HzR: 6.44, 95% CI: 1.44‐28.86), heart rate (HR) < 100 beats per minute (bpm; HzR: 12.2; 95% CI: 2.54‐58.62), HR > 140 bpm (HzR: 3.59, 95% CI: 1.05‐12.33), neutrophil count <1.7 × 109/L (HzR: 7.36; 95% CI: 2.03‐26.66) and increased gamma‐glutamyl transferase activity (every 50‐unit, HzR: 1.12; 95% CI: 1.03‐1.21) were predictive of nonsurvival. Conclusions and Clinical Importance: The use of Hp and GLDH for prediction of survival in sick calves cannot be recommended at this time. [ABSTRACT FROM AUTHOR]

The article focuses on the release of the Don Carlos INTA peach by the San Pedro Experiment Station, Instituto Nacional de Tecnología Agropecuaria (INTA) in Buenos Aires, Argentina. The Don Carlos INTA is said to provide a medium-chilling, commercially acceptable peach that ripens after the peaches June Gold and Ginart. It details the origin of the Don Carlos INTA. It describes the fruit of the Don Carlos INTA adult trees.

The article presents a letter to the editor about Clostridium difficile-associated disease.

A letter to the editor is presented in response to the article "Effects of Alkalinisation and Rehydration on Plasma Potassium Concentration in Nenonatal Calves with Diarrhea," by F. M. Trefz et al. in the March 2015 issue.

OBJECTIVE: To describe histologic findings in the small airways during postmortem examination of actively racing horses and to quantify the degree of airway inflammation by use of a semiquantitative scoring system. SAMPLE: Lung tissues obtained from 95 horses (Thoroughbreds, Standardbreds, and Quarter Horses) that had been actively racing or training. PROCEDURES: 10 standardized lung sections were obtained during postmortem examination of 37 horses that had been actively racing or training, and 2 histologic sections of the caudodorsal lung lobes previously collected from each of 58 horses that died or were euthanized during racing or training were also obtained. Sections were evaluated by use of a validated histologic small airway scoring system. RESULTS: Scores for inflammatory cells, smooth muscle, and hemosiderin typically were high. Signalment and cause of death were not significant predictors of lung scores. Lung sample location was a significant predictor, with the highest scores in the caudal and dorsal sections. Inflammatory cell infiltration in peribronchiolar tissues, smooth muscle hyperplasia, and hemosiderin (prevalence of 86%, 98%, and 80%, respectively) were common findings in lungs of these horses, with the caudodorsal regions more severely affected. Correlation was moderate between smooth muscle hyperplasia and inflammatory cell infltration, with minimal correlation between hemosiderin and inflammatory cell infiltration. CONCLUSIONS AND CLINICAL RELEVANCE: Inflammatory airway disease has been identified by use of bronchoalveolar lavage in young athletic horses throughout the world. In the study reported here, pathological changes were detected in the wall of small airways of horses that were actively training or racing. [ABSTRACT FROM AUTHOR]

A case study involves a 2-day-old Appaloosa filly presenting with straining and abdominal distension. Diagnostic imaging via abdominal ultrasonography reveals a large amount of homogeneous anechoic free peritoneal fluid with aggregation of the intestinal tube, indicative of uroperitoneum. Differential diagnoses include uroperitoneum, Foley, neonatal foal, medical management and equine.

Gunshot residue (GSR) is one of the few forensic disciplines that lack accurate screening techniques. This study proposes using a mobile LIBS instrument to detect inorganic GSR and compares performance to a previously validated laboratory instrument. The mobile LIBS is designed with advanced configurations specifically for on-site GSR analysis, including a CMOS detector and a sampling chamber that holds up to six typical GSR collection devices with separate gas flow ports to prevent cross-contamination. A significant novelty of the portable instrument is its image magnification, which allows quick searching and visualization of GSR particle morphology. The single-particle imaging and elemental composition capability is one of a kind and offers superior confirmatory features for GSR. The mobile LIBS performance was evaluated for residues collected from the hands of shooters (100 samples) and non-shooters (200 background samples), analyzed sequentially by the mobile instrument and then the laboratory instrument. Accuracies better than 98.8% were obtained by both instruments, demonstrating their suitability for trace IGSR detection from skin specimens. Implementation of this methodology is anticipated to drastically speed up response times (i.e., from several hours per sample by standard SEM-EDS practice to a few minutes by LIBS). The screening methods can be easily incorporated into workflows to improve decision-making processes at the crime scene and laboratory settings, reduce backlogs, and improve case management. [Display omitted] • Evolution of laboratory LIBS analysis to in-field GSR LIBS analysis. • Improved magnification allowed novel micron-GSR searching on mobile LIBS. • Instrument performances tested on 300 authentic shooter GSR hand specimens. • Over 98% accuracy for both instruments, increasing validity of field LIBS screening. [ABSTRACT FROM AUTHOR]

Neurotoxic side effects of chemotherapy include deficits in attention, memory, and executive functioning. Currently, there are no FDA-approved therapies. In mice, cisplatin causes long-term cognitive deficits, white matter damage, mitochondrial dysfunction, and loss of synaptic integrity. We hypothesized that MSC-derived small extracellular vesicles (sEVs) could restore cisplatin-induced cognitive impairments and brain damage. Animals were injected with cisplatin intraperitoneally and treated with MSC-derived sEVs intranasally 48 and 96 h after the last cisplatin injection. The puzzle box test (PBT) and the novel object place recognition test (NOPRT) were used to determine cognitive deficits. Synaptosomal mitochondrial morphology was analyzed by transmission electron microscopy. Immunohistochemistry using antibodies against synaptophysin and PSD95 was applied to assess synaptic loss. Black-Gold II staining was used to quantify white matter integrity. Our data show that sEVs enter the brain in 30 min and reverse the cisplatin-induced deficits in executive functioning and working and spatial memory. Abnormalities in mitochondrial morphology, loss of white matter, and synaptic integrity in the hippocampus were restored as well. Transcriptomic analysis revealed upregulation of regenerative functions after treatment with sEVs, pointing to a possible role of axonal guidance signaling, netrin signaling, and Wnt/Ca2+ signaling in recovery. Our data suggest that intranasal sEV treatment could become a novel therapeutic approach for the treatment of chemobrain. [ABSTRACT FROM AUTHOR]

Next generation sequencing has demonstrated that alpha diversity of the fecal microbiota is significantly altered in horses with typhlocolitis. The objective of this study was to evaluate the bacterial composition of the cecum content of horses with and without typhlocolitis through direct and culture-enriched 16S gene sequencing of six healthy horses and six horses with acute typhlocolitis; a case-control study design. Cecal content was collected after euthanasia. An aliquot was used for direct 16S gene sequencing. Another was serially diluted with brain heart infusion (BHI) and plated onto five different agar media. All culture medias, except for MacConkey, were incubated anaerobically. Bacterial colonies were harvested in bulk and used for DNA extraction, 16S PCR amplification, and sequenced using the Illumina MiSeq platform. Predominant phyla in healthy and diseased horses were Firmicutes, followed by Bacteroidetes in all cultured medias, except for MacConkey agar, in which Proteobacteria was the dominant phylum. Greater bacterial richness was identified in sequenced cecal contents as compared to cultured plates (P < 0.05). Culture-enriched molecular profiling combined with 16S rRNA gene sequencing offer an alternative method for the study of the gut microbiota of horses. For direct cecum content 16S gene amplification, the alpha diversity indices were lower in diarrheic horses compared to healthy horses (P < 0.05). A higher relative abundance of Fusobacteriota was found in 2/6 samples from diarrheic horses. The role of Fusobacteriota in equine colitis deserves investigation. [ABSTRACT FROM AUTHOR]

A letter to the editor is presented in response to the article "Hospitalized community-acquired pneumonia due to Streptococcus pneumoniae: has resistance to antibiotics decreased?," by Ignacio Obando and Luis Arroyo published in a previous issue of the periodical.

Alveolar macrophages (AMs) are the predominant innate immune cell in the distal respiratory tract. During inflammatory responses, AMs may be supplemented by blood monocytes, which differentiate into monocyte-derived macrophages (MDMs). Macrophages play important roles in a variety of common equine lower airway diseases, including severe equine asthma (SEA). In an experimental model, an inhaled mixture of Aspergillus fumigatus spores, lipopolysaccharide, and silica microspheres (FLS), induced SEA exacerbation in susceptible horses. However, whether equine AMs and MDMs have differing immunophenotypes and cytokine responses to FLS stimulation is unknown. To address these questions, alveolar macrophages/monocytes (AMMs) were isolated from bronchoalveolar lavage fluid and MDMs derived from blood of six healthy horses. Separately, AMMs and MDMs were cultured with and without FLS for six hours after which cell surface marker expression and cytokine production were analyzed by flow cytometry and a bead-based multiplex assay, respectively. Results showed that regardless of exposure conditions, AMMs had significantly higher surface expression of CD163 and CD206 than MDMs. Incubation with FLS induced secretion of IL-1β, IL-8, TNF-α and IFN-γ in AMMs, and IL-8, IL-10 and TNF-α in MDMs. These results suggest that AMMs have a greater proinflammatory response to in vitro FLS stimulation than MDMs, inferring differing roles in equine lung inflammation. Variability in recruitment and function of monocyte-macrophage populations warrant more detailed in vivo investigation in both homeostatic and diseased states. [ABSTRACT FROM AUTHOR]

Background: The intestinal tract is a rich and complex environment and its microbiota has been shown to have an important role in health and disease in the host. Several factors can cause disruption of the normal intestinal microbiota, including antimicrobial therapy, which is an important cause of diarrhea in horses. This study aimed to characterize changes in the fecal bacterial populations of healthy horses associated with the administration of frequently used antimicrobial drugs. Results: Twenty-four adult mares were assigned to receive procaine penicillin intramuscularly (IM), ceftiofur sodium IM, trimethoprim sulfadiazine (TMS) orally or to a control group. Treatment was given for 5 consecutive days and fecal samples were collected before drug administration (Day 1), at the end of treatment (Days 5), and on Days 14 and 30 of the trial. High throughput sequencing of the V4 region of the 16S rRNA gene was performed using an Illumina MiSeq sequencer. Significant changes of population structure and community membership were observed after the use of all drugs. TMS caused the most marked changes on fecal microbiota even at higher taxonomic levels including a significant decrease of richness and diversity. Those changes were mainly due to a drastic decrease of Verrucomicrobia, specifically the "5 genus incertae sedis". Changes in structure and membership caused by antimicrobial administration were specific for each drug and may be predictable. Twenty-five days after the end of treatment, bacterial profiles were more similar to pre-treatment patterns indicating a recovery from changes caused by antimicrobial administration, but differences were still evident, especially regarding community membership. Conclusions: The use of systemic antimicrobials leads to changes in the intestinal microbiota, with different and specific responses to different antimicrobials. All antimicrobials tested here had some impact on the microbiota, but TMS significantly reduced bacterial species richness and diversity and had the greatest apparent impact on population structure, specifically targeting members of the Verrucomicrobia phylum. [ABSTRACT FROM AUTHOR]

Analysis of gunshot residue currently lacks effective screening methods that can be implemented in real time at the crime scene. Historically, SEM‐EDS has been the standard for analysis; however, advances in technology have brought portable instrumentation to the forefront of forensic science disciplines, including the screening of GSR. This study proposes electrochemical methods with disposable screen‐printed carbon electrodes for GSR screening at the laboratory and points of care due to their rapid, cost‐efficient, and compact platform. GSR residues were extracted from typical aluminum/carbon adhesive collection stubs and analyzed via square‐wave anodic stripping voltammetry. Benchtop and portable electrochemical instruments were compared for the assessment and classification of authentic shooter samples by monitoring a panel of inorganic and organic GSR elements and compounds including lead, antimony, copper, 2,4‐dinitrotoluene, diphenylamine, nitroglycerin, and ethyl centralite. The evaluation included the assessment of figures of merit and performance measures from quality controls, nonshooter, and shooter data sets. Samples collected from the hands of 200 background individuals (nonshooters), and shooters who fired leaded ammunition (100) and lead‐free ammunition (50) were analyzed by the benchtop and portable systems with accuracies of 95.7% and 96.5%, respectively. The findings indicate that electrochemical methods are fast, sensitive, and specific for the identification of inorganic and organic gunshot residues. The portable potentiostat provided results comparable with the benchtop system, serving as a proof‐of‐concept to transition this methodology to crime scenes for a practical and inexpensive GSR screening that could reduce backlogs, improve investigative leads, and increase the impact of gunshot residues in forensic science. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the fecal shedding of C. difficile in calves on farms in Sao Paulo State, Brazil. Fecal samples (n = 300) were collected from diarrheic (n = 78) and nondiarrheic (n = 222) calves less than 60 days of age from 20 farms. Fecal samples were inoculated into enrichment broth supplemented with taurocholate and cultured under anaerobic conditions. Colonies suspected to be C. difficile were harvested for DNA extraction and then multiplex PCR for the detection of genes encoding toxins A and B and binary toxins. All toxigenic isolates were ribotyped and tested for antimicrobial susceptibility, and five selected strains were subjected to whole-genome sequencing to determine their sequence type. C. difficile was isolated from 29.3 % (88/300) of the samples. All toxigenic isolates (17/88, 19.3 %) were classified as ribotypes RT046 (13/17–79.47 %, A+B+ CDT−) and RT126 (4/17 = 20.53 %, A+B+ CDT+). The sequenced strains from RT046 were classified as ST35 (Clade 1), while those from RT126 were classified as ST11 (Clade 5). No associations between the epidemiological factors in any of the groups and C. difficile isolation were observed. Most of the toxigenic isolates (16/17 = 94.41 %) were classified as multidrug-resistant. Calves can be an important source of toxigenic C. difficile strains, including multidrug-resistant isolates from ribotypes commonly observed in humans. • C. difficile was isolated from fecal samples of calves with and without diarrhea. • Toxigenic isolates were classified as RT046 and RT126. • Most of the toxigenic isolates were classified as multidrug resistant. [ABSTRACT FROM AUTHOR]

Objective--To determine effects of extracorporeal shock wave therapy (ESWT) on healing of wounds in the distal portion of the forelimb in horses. Animals-6 horses. Procedures--Five 6.25-cm² superficial wounds were created over both third metacarpi of 6 horses. Forelimbs were randomly assigned to treatment (ESWT and bandage) or control (bandage only) groups. In treated limbs, each wound was treated with 625 shock wave pulses from an unfocused electrohydraulic shock wave generator. In control limbs, each wound received sham treatment. Wound appearance was recorded weekly as inflamed or healthy and scored for the amount of protruding granulation tissue. Standardized digital photographs were used to determine the area of neoepithelialization and absolute wound area. Biopsy was performed on 1 wound on each limb every week for 6 weeks to evaluate epithelialization, fibroplasia, neovascularization, and inflammation. Immunohistochemical staining fora smooth muscle actin was used to label myofibroblasts. Results--Control wounds were 1.9 times as likely to appear inflamed, compared with treated wounds. Control wounds had significantly higher scores for exuberant granulation tissue. Treatment did not affect wound size or area of neoepithelialization. No significant difference was found for any of the histologic or immunohistochemical variables between groups. Conclusions and Clinical Relevance--Treatment with ESWT did not accelerate healing of equine distal limb wounds, but treated wounds had less exuberant granulation tissue and appeared healthier than controls. Therefore, ESWT may be useful to prevent exuberant granulation tissue formation and chronic inflammation of such wounds, but further studies are necessary before recommending ESWT for clinical application. [ABSTRACT FROM AUTHOR]

Pediatric parapneumonic empyema (PPE) has been increasing in several countries including Spain. Streptococcus pneumoniae is a major PPE pathogen; however, antimicrobial pretreatment before pleural fluid (PF) sampling frequently results in negative diagnostic cultures, thus greatly underestimating the contribution of pneumococci, especially pneumococci susceptible to antimicrobial agents, to PPE. The study aim was to identify the serotypes and genotypes that cause PPE by using molecular diagnostics and relate these data to disease incidence and severity. A total of 208 children with PPE were prospectively enrolled; blood and PF samples were collected. Pneumococci were detected in 79% of culture-positive and 84% of culture-negative samples. All pneumococci were genotyped by multilocus sequence typing. Serotypes were determined for 111 PPE cases; 48% were serotype 1, of 3 major genotypes previously circulating in Spain. Variance in patient complication rates was statistically significant by serotype. The recent PPE increase is principally due to nonvaccine serotypes, especially the highly invasive serotype 1. [ABSTRACT FROM AUTHOR]

Abstract: Clostridium difficile is the most common cause of hospital- and antimicrobial-associated diarrhea in hospitalized humans however the role of C. difficile in diarrhea in dogs has not been defined. A prospective study of C. difficile colonization in dogs and cats was conducted in a veterinary teaching hospital intensive care unit (ICU). Rectal swabs were taken from patients upon admission to the ICU and every third day of hospitalization until discharge or death. C. difficile was isolated from 73/402 (18%) animals; 69% of isolates were toxigenic. Community-associated colonization was identified in 39/366 (11%) of animals that were sampled at the time of admission, while C. difficile was subsequently isolated from 27 of the remaining 327 (8.3%) animals that had a negative admission swab. C. difficile was isolated from seven other dogs during hospitalization, but the origin was unclear because the admission swab was not collected. Administration of antimicrobials prior to admission and administration of immunosuppressive drugs during hospitalization were risk factors for hospital-associated colonization (P =0.006, OR 4.05, 95% CI 1.4–10.8). Acquisition of C. difficile during hospitalization in the ICU was associated with the development of diarrhea (P =0.004). Two ribotypes, one toxigenic and one non-toxigenic, predominated. [Copyright &y& Elsevier]

Background: Prior retrospective studies have suggested that tacrolimus monotherapy is an option associated with excellent outcomes and reduced toxicities. Method: We conducted a prospective, randomized, 2-center study of tacrolimus combination therapy vs monotherapy. From April 16, 2004, to September 15, 2005, 58 adult heart transplant patients were studied. All received oral tacrolimus, mycophenolate mofetil, and corticosteroids. Patients were then randomized to a group where mycophenolate was maintained (COMBO) or to a group where it was discontinued (MONO) 14 days post-transplant. Corticosteroids were rapidly withdrawn in both groups between 8 and 12 weeks. Result: The primary end point (mean 6-month International Society of Heart and Lung Transplantation biopsy score) was 0.44 ± 0.04 in the MONO group and 0.60 ± 0.05 in the COMBO group (p = 0.013, unpaired Student’s t-test). The freedom from rejection grade of 2R or higher at 6 and 12 months was 93.3% with MONO and 92.9% with COMBO (p = NS). Conclusion: Tacrolimus monotherapy appears to be safe and efficacious in heart transplant recipients and is not associated with excess rejection in the first year post-transplant. Further studies of this approach are warranted. [Copyright &y& Elsevier]

Background: There is a paucity of knowledge on the long-term outcome in patients diagnosed with COVID-19. We describe a cohort of patients with a constellation of symptoms occurring four weeks after diagnosis causing different degrees of reduced functional capacity. Although different hypothesis have been proposed to explain this condition like persistent immune activation or immunological dysfunction, to date, no physiopathological mechanism has been identified. Consequently, there are no therapeutic options besides symptomatic treatment and rehabilitation.Methods: We evaluated patients with symptoms that persisted for at least 4 weeks after COVID-19. Epidemiological and clinical data were collected. Blood tests, including inflammatory markers, were conducted, and imaging studies made if deemed necessary. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) reverse transcription polymerase chain reaction (RT-PCR) in plasma, stool, and urine were performed. Patients were offered antiviral treatment (compassionate use).Results: We evaluated 29 patients who reported fatigue, muscle pain, dyspnea, inappropriate tachycardia, and low-grade fever. Median number of days from COVID-19 to positive RT-PCR in extra-respiratory samples was 55 (39-67). Previous COVID-19 was mild in 55% of the cases. Thirteen patients (45%) had positive plasma RT-PCR results and 51% were positive in at least one RT-PCR sample (plasma, urine, or stool). Functional status was severely reduced in 48% of the subjects. Eighteen patients (62%) received antiviral treatment. Improvement was seen in most patients (p = 0.000) and patients in the treatment group achieved better outcomes with significant differences (p = 0.01).Conclusions: In a cohort of COVID-19 patients with persistent symptoms, 45% of them have detectable plasma SARS-CoV-2 RNA. Our results indicate possible systemic viral persistence in these patients, who may benefit of antiviral treatment strategies. [ABSTRACT FROM AUTHOR]

To assess cardiopulmonary function in sedated and anesthetized dogs administered intravenous (IV) dexmedetomidine and subsequently administered IV lidocaine to treat dexmedetomidine-induced bradycardia. Prospective, randomized, crossover experimental trial. A total of six purpose-bred female Beagle dogs, weighing 9.1 ± 0.6 kg (mean ± standard deviation). Dogs were randomly assigned to one of three treatments: dexmedetomidine (10 μg kg–1 IV) administered to conscious (treatments SED1 and SED2) or isoflurane-anesthetized dogs (end-tidal isoflurane concentration 1.19 ± 0.04%; treatment ISO). After 30 minutes, a lidocaine bolus (2 mg kg–1) IV was administered in treatments SED1 and ISO, followed 20 minutes later by a second bolus (2 mg kg–1) and a 30 minute lidocaine constant rate infusion (L-CRI) at 50 (SED1) or 100 μg kg–1 minute–1 (ISO). In SED2, lidocaine bolus and L-CRI (50 μg kg–1 minute–1) were administered 5 minutes after dexmedetomidine. Cardiopulmonary measurements were obtained after dexmedetomidine, after lidocaine bolus, during L-CRI and 30 minutes after discontinuing L-CRI. A mixed linear model was used for comparisons within treatments (p < 0.05). When administered after a bolus of dexmedetomidine, lidocaine bolus and L-CRI significantly increased heart rate and cardiac index, decreased mean blood pressure, systemic vascular resistance index and oxygen extraction ratio, and did not affect stroke volume index in all treatments. Lidocaine was an effective treatment for dexmedetomidine-induced bradycardia in healthy research dogs. [ABSTRACT FROM AUTHOR]

• Multiple cases of non-biologic-associated Theiler's disease occurred on a farm in Ontario, Canada in 2005. • Retrospective analysis of serum showed the presence of EqPV-H DNA in horses with a confirmed histological diagnosis of Theiler's disease. • First report of the association of EqPV-H with Theiler's disease in Canada. • The highest EqPV-H DNA viral load was detected in a clinically normal animal • Phylogenetic analysis of EqPV-H showed high similarity to previous published sequences. Theiler's disease was confirmed within a group horses located on a farm in southwestern Ontario during the summer and autumn of 2005. Five sudden deaths occurred between 3 July and 21 August, 2005, none of which were necropsied, however two of the horses showed clinical signs compatible with hepatic encephalopathy prior to death. No horse on the farm had received a biologic product of equine blood origin in the preceding six months. The only biologics used on the property were the administration of killed vaccines for rabies, tetanus and West Nile Virus to all horses 30 days prior to the onset of the first sudden death. Between 22 August, 2005 and 21 October, 2005, a further four horses died suddenly or were euthanized with all having a confirmed histopathologic diagnosis of acute hepatic necrosis. Serum was collected from all horses on the farm on 30 September, 2005 and this was repeated on 29 October, 2005. Equine parvovirus-hepatitis (EqPV-H) DNA was detected by quantitative-PCR in the serum of 61.8% (34/55) of the horses on the farm on either one or both sampling dates with viral loads ranging from <3.75 × 103 copies/mL to 3.64 × 107 copies/mL. EqPV-H DNA was present in serum samples of three horses with a confirmed diagnosis of Theiler's disease, five horses with subclinical liver disease, and in clinically normal in-contact horses. Subsequent phylogenetic analysis based on partial NS1 of EqPV-H revealed not only high similarity on nucleotide level within the sequenced samples but also within other previously published sequences. [ABSTRACT FROM AUTHOR]

The increasing demand for rapid methods to identify both inorganic and organic gunshot residues (IGSR and OGSR) makes electrochemical methods, an attractive screening tool to modernize current practice. Our research group has previously demonstrated that electrochemical screening of GSR samples delivers a simple, inexpensive, and sensitive analytical solution that is capable of detecting IGSR and OGSR in less than 10 min per sample. In this study, we expand our previous work by increasing the number of GSR markers and applying machine learning classifiers to the interpretation of a larger population data set. Utilizing bare screen‐printed carbon electrodes, the detection and resolution of seven markers (IGSR; lead, antimony, and copper, and OGSR; nitroglycerin, 2,4‐dinitrotoluene, diphenylamine, and ethyl centralite) was achieved with limits of detection (LODs) below 1 µg/mL. A large population data set was obtained from 395 authentic shooter samples and 350 background samples. Various statistical methods and machine learning algorithms, including critical thresholds (CT), naïve Bayes (NB), logistic regression (LR), and neural networks (NN), were utilized to calculate the performance and error rates. Neural networks proved to be the best predictor when assessing the dichotomous question of detection of GSR on the hands of shooter versus nonshooter groups. Accuracies for the studied population were 81.8 % (CT), 88.1% (NB), 94.7% (LR), and 95.4% (NN), respectively. The ability to detect both IGSR and OGSR simultaneously provides a selective testing platform for gunshot residues that can provide a powerful field‐testing technique and assist with decisions in case management. [ABSTRACT FROM AUTHOR]

Prior to Hurricane Maria, Puerto Rico already had 200+ hazardous waste sites, significant contamination of water resources, and among the highest rates of preterm birth in the USA. To address these issues, the Puerto Rico Testsite for Exploring Contamination Threats (PROTECT) Center was formed in 2010 to investigate prenatal environmental exposures, particularly phthalates, and adverse birth outcomes. Recent work from the PROTECT study confirms that in utero exposure to certain phthalates is associated with shorter gestation and increased risk of preterm birth. However, previous research also suggests that pregnant women who experience a natural disaster such as Hurricane Maria are at higher risk of adverse birth outcomes, but it is unknown whether this is due to stress, hazardous exposures, or a combination of factors. Thus, the aim of this analysis was to characterize hurricane-related changes in phthalate exposures and experiences within the PROTECT cohort. Among 176 participants who were pregnant during or within 5 months after Maria, 122 completed a questionnaire on hurricane-related experiences. Questionnaire results and biomarkers of exposure suggest that participants did not have regular access to fresh foods and water during hurricane recovery, and almost half reported structural damage to their home. In addition, biomarker concentrations of phthalates commonly used in food packaging were higher among participants post-hurricane, while phthalates commonly used in personal care products were lower compared to pre-hurricane levels. Hurricane-related increases in phthalate exposure, as well as widespread structural damage, food and water shortages, and long-term absence of electricity and cell phone service, likely increased the risk of adverse birth outcomes among this already vulnerable population. [ABSTRACT FROM AUTHOR]

Introduction: In the equine racehorse industry, reduced athletic performance due to joint injury and lameness has been extensively reviewed. Intra-articular injections of glucocorticoids are routinely used to relieve pain and inflammation associated with osteoarthritis. Intra-articular injections of pharmaceutical agents require practice for precise needle placement and to minimize complications. Training on simulators or models is a viable alternative for developing these technical skills. The purpose of this study was to compare the qualitative ultrasonographic characteristics of three-dimensional (3D) printed models of equine cervical articular process joints to that of a dissected equine cervical spine (gold standard). Methods: A randomized complete block design study was conducted in which a total of thirteen cervical articular process joint models were printed using several materials, printers, and printing technologies. Ultrasound video clips with the models immersed in water were recorded. Two board certified veterinary radiologists and three veterinary radiology residents reviewed the videos and responded to a survey assessing and comparing the ultrasonographic characteristics of the 3D printed models to those of the gold standard. Results: Six 3D printed models had ultrasonographic characteristics similar to the gold standard. These six models were (material, printer, printing technology): nylon PA 12, EOS Formiga P100, selective laser sintering (P = 0.99); Onyx nylon with chopped carbon fiber, Markforged Onyx Two, fused deposition modeling (P = 0.48); polycarbonate, Ultimaker 3, fused deposition modeling (P = 0.28); gypsum, ProJet CJP 660 Pro, ColorJet Printing (P = 0.28); polylactic acid, Prusa I3, fused deposition modeling (P = 0.23); and high temperature V1 resin, Form 2, stereolithography (P = 0.22). Conclusion: When assessed in water, it is possible to replicate the qualitative ultrasonographic characteristics of bone using three dimensional printed models made by combining different materials, printing technologies, and printers. However, not all models share similar qualitative ultrasonographic characteristics with bone. We suggest that the aforementioned six models be used as proxy for simulating bones or joints for use with ultrasound. In order to replicate the resistance and acoustic window provided by soft tissues, further work testing the ability of these models to withstand embedding in material such as ballistic gelatin is required. [ABSTRACT FROM AUTHOR]

Highlights • Non-parametric was more informative than parametric ANOVA. • Test based on mixed model plus interaction-wise is recommended. • SREG was more useful than AMMI models for studying pattern interaction. Abstract The presence of genotype-by-environment interactions (GE) remains a major issue for crop improvement. The aims of this work were: i) to compare the efficiency of parametric and non-parametric methods to test the presence of crossover (COI) and non-crossover GE (NCOI), ii) visual examination of the relationships between environments and genotypes tested, and iii) to test the effectiveness of dividing the peach season evaluations into mega-environments (ME) using the biplot based on AMMI and SREG. Non-parametric ANOVA was more useful than the parametric approach because it can distinguish between the presence of COI and NCOI. Three test methods, suitable for investigating two-factor interactions, were used to show that interactions between genotypes and environment involve significant changes in rank order. The Yang test based on mixed model theory combined with interaction-wise error rate was the most sensitive to detect COI, while the Gail and Simon, as well as the Azzalini and Cox methods were conservative. Which-won-where pattern was followed with four and two ME were found with AMMI and SREG, respectively. Entries G16 (Hermosillo P), G21 (María Emilia N), G2 (84.351.029 N) and G8 (Cotogna del Berti P) showed specific adaptability to ME-1, ME-2, ME-3 and ME-4 generated by AMMI, respectively; while G28 (Sunprince P) exhibited specific adaptation to ME-1 and G16 in ME-2 which were created by SREG. Average environment coordination (AEC) view of the GGE biplot involving the seven environments identified G10 (Flameprince P) as the most stable and high-yielding genotype across environments, unlike G8 and G28, which showed only high yields. Results indicated that AMMI and GGE biplots are informative methods to explore stability and adaptation patterns of genotypes in practical plant breeding and in subsequent variety recommendations. In addition, finding ME helps identify the most suitable peach genotypes that can be recommended for areas within a specific ME in either one or more test locations. [ABSTRACT FROM AUTHOR]