The city nature challenge: A global citizen science phenomenon contributing to biodiversity knowledge and informing local government practices.

The bioblitz phenomenon has recently branched into cities, presenting exciting opportunities for local governments to channel participants' efforts toward local issues. The City Nature Challenge (CNC) is one such initiative that has been quickly taken up by hundreds of municipalities worldwide. Despite high levels of participation, we still lack a framework for evaluating how the CNC contributes to local biodiversity knowledge and to inform local government practices. In the present article, we develop such a tool and present a case study that illustrates its applicability. We demonstrate that the collected records contributed to a better understanding of contemporary, local biodiversity patterns and provide a more realistic representation of understudied groups such as insects and fungi. Importantly, we show that the CNC presented local governments with a cost-effective tool to make informed, evidence-based management and policy decisions, improve education and engagement programs, foster cross-council collaborations, and support a stronger sense of environmental stewardship within the local community.

Preoperative Atelectasis in Patients with Obesity Undergoing Bariatric Surgery: A Cross-Sectional Study.

BACKGROUND: Pulmonary atelectasis is present even before surgery in patients with obesity. We aimed to estimate the prevalence and extension of preoperative atelectasis in patients with obesity undergoing bariatric surgery and to determine if variation in preoperative Spo2 values in the seated position at room air is explained by the extent of atelectasis coverage in the supine position. METHODS: This was a cross-sectional study in a single center specialized in laparoscopic bariatric surgery. Preoperative chest computed tomographies were reassessed by a senior radiologist to quantify the extent of atelectasis coverage as a percentage of total lung volume. Patients were classified as having atelectasis when the affection was ≥2.5%, to estimate the prevalence of atelectasis. Crude and adjusted prevalence ratios (aPRs) and odds ratios (aORs) were obtained to assess the relative prevalence of atelectasis and percentage coverage, respectively, with increasing obesity category. Inverse probability weighting was used to assess the total, direct (not mediated), and indirect (mediated through atelectasis) effects of body mass index (BMI) on preoperative Spo2, and to quantify the magnitude of mediation (proportion mediated). E-values were calculated, to represent the minimum magnitude of association that an unmeasured confounder with the same directionality of the effect should have to drive the observed point estimates or lower confidence intervals (CIs) to 1, respectively. RESULTS: In 236 patients with a median BMI of 40.3 kg/m2 (interquartile range [IQR], 34.6-46.0, range: 30.0-77.3), the overall prevalence of atelectasis was 32.6% (95% CI, 27.0-38.9) and by BMI category: 30 to 35 kg/m2, 12.7% (95% CI, 6.1-24.4); 35 to 40 kg/m2, 28.3% (95% CI, 17.2-42.6); 40 to 45 kg/m2, 12.3% (95% CI, 5.5-24.3); 45 to 50 kg/m2, 48.4% (95% CI, 30.6-66.6); and ≥50 units, 100% (95% CI, 86.7-100). Compared to the 30 to 35 kg/m2 group, only the categories with BMI ≥45 kg/m2 had significantly higher relative prevalence of atelectasis-45 to 50 kg/m2, aPR = 3.52 (95% CI, 1.63-7.61, E-value lower bound: 2.64) and ≥50 kg/m2, aPR = 8.0 (95% CI, 4.22-15.2, E-value lower bound: 7.91)-and higher odds of greater atelectasis percentage coverage: 45-50 kg/m2, aOR = 7.5 (95% CI, 2.7-20.9) and ≥50 kg/m2, aOR = 91.5 (95% CI, 30.0-279.3). Atelectasis percent alone explained 70.2% of the variation in preoperative Spo2. The proportion of the effect of BMI on preoperative Spo2 values <96% mediated through atelectasis was 81.5% (95% CI, 56.0-100). CONCLUSIONS: The prevalence and extension of atelectasis increased with higher BMI, being significantly higher at BMI ≥45 kg/m2. Preoperative atelectasis mediated the effect of BMI on Spo2 at room air in the seated position.

Indigenous plants promote insect biodiversity in urban greenspaces.

The contribution of urban greenspaces to support biodiversity and provide benefits for people is increasingly recognized. However, ongoing management practices favor vegetation oversimplification, often limiting greenspaces to lawns and tree canopy rather than multi-layered vegetation that includes under- and midstorey, and the use of nonnative species. These practices hinder the potential of greenspaces to sustain indigenous biodiversity, particularly for taxa like insects that rely on plants for food and habitat. Yet, little is known about which plant species may maximize positive outcomes for taxonomically and functionally diverse insect communities in greenspaces. Additionally, while cities are expected to experience high rates of introductions, quantitative assessments of the relative occupancy of indigenous vs. introduced insect species in greenspace are rare, hindering understanding of how management may promote indigenous biodiversity while limiting the establishment of introduced insects. Using a hierarchically replicated study design across 15 public parks, we recorded occurrence data from 552 insect species on 133 plant species, differing in planting design element (lawn, midstorey, and tree canopy), midstorey growth form (forbs, lilioids, graminoids, and shrubs) and origin (nonnative, native, and indigenous), to assess (1) the relative contributions of indigenous and introduced insect species and (2) which plant species sustained the highest number of indigenous insects. We found that the insect community was overwhelmingly composed of indigenous rather than introduced species. Our findings further highlight the core role of multi-layered vegetation in sustaining high insect biodiversity in urban areas, with indigenous midstorey and canopy representing key elements to maintain rich and functionally diverse indigenous insect communities. Intriguingly, graminoids supported the highest indigenous insect richness across all studied growth forms by plant origin groups. Our work highlights the opportunity presented by indigenous understory and midstorey plants, particularly indigenous graminoids, in our study area to promote indigenous insect biodiversity in urban greenspaces. Our study provides a blueprint and stimulus for architects, engineers, developers, designers, and planners to incorporate into their practice plant species palettes that foster a larger presence of indigenous over regionally native or nonnative plant species, while incorporating a broader mixture of midstorey growth forms.

Critical analysis and limitations of resting ankle-brachial index in the diagnosis of symptomatic peripheral arterial disease patients and the role of diabetes mellitus and chronic kidney disease.

BACKGROUND: The ankle-brachial index (ABI) may underestimate the severity of peripheral arterial disease (PAD) in patients with noncompressible vessels. This study analyzed limitations of the ABI and toe-brachial index (TBI), if done alone, in patients with symptomatic PAD, diagnosed by duplex ultrasound (DUS) examination, particularly in patients with diabetes and chronic kidney disease (CKD). METHODS: This is a retrospective review of prospectively collected data. All patients underwent resting ABIs, TBI, and/or DUS. An ABIs of 0.90 or less in either leg was considered abnormal, and the term inconclusive ABIs (noncompressibility) was used if the ABI was 1.3 or greater. The sensitivity, specificity, positive predictive value, negative predictive value, and overall accuracy (OA) of ABIs in detecting 50% or greater stenosis of any arterial segment based on DUS were determined. A TBI of less than 0.7 was considered abnormal. RESULTS: We included 2226 ABIs and 1383 DUS examinations: 46% of patients had diabetes, 16% had CKD, and 39% had coronary artery disease. Fifty-three percent of the ABIs were normal, 34% were abnormal, and 13% were inconclusive. For patients with limb-threatening ischemia, 40% had normal ABIs, 40% abnormal ABIs, and 20% were inconclusive. The sensitivity and OA for ABIs in detecting 50% or greater stenosis in the whole series were 57% (95% confidence interval [CI], 53.7-61.2) and 74% (95% CI, 71.9-76.6); for diabetics 51% (95% CI, 46.1-56.3) and 66% (95% CI, 62.3-69.8); nondiabetics 66% (95% CI, 59.9-70.9) and 81% (95% CI, 78.2-83.9). For patients with CKD, the sensitivity and OA for ABIs in detecting 50% or greater stenosis was 43% (95% CI, 34.3-52.7) and 67% (95% CI, 60.2-73.0) versus patients with no CKD 60% (95% CI, 56.3-64.6) and 76% (95% CI, 73.1-78.1). If patients with inconclusive ABIs were excluded, these values were 69% (95% CI, 65.2-72.9) and 80% (95% CI, 77.2-81.9) in the whole series; 67% (95% CI, 61.6-72.7) and 75% (95% CI, 70.5-78.4) for diabetics; and 63% (95% CI, 51.3-73.0) and 78% (95% CI, 70.6-83.9) for patients with CKD. Thirty-three percent of TBIs were normal and 67% were abnormal. The sensitivity and OA for abnormal TBI in detecting 50% or greater stenosis were 85% (95% CI, 78.9-90.0) and 75% (95% CI, 70.1-80.2) in the whole series; 84% (95% CI, 76.0-90.3) and 74% (95% CI, 67.1-80.2) for diabetics; and 77% (95% CI, 61.4-88.2) and 72% (95% CI, 59.9-82.3) for patients with CKD. For those with inconclusive ABIs, these values for TBI were 75% and 69%. CONCLUSIONS: Of symptomatic patients with PAD with 50% or greater stenosis on DUS examination, 43% had normal/inconclusive resting ABIs (49% in diabetics and 57% in CKD). TBI may help in patients with inconclusive ABIs. These patients should undergo further imaging to determine proper treatment.

Exogenous melatonin and male foetuses improve the quality of sheep colostrum.

Colostrum is the first product secreted by the mammary gland to transfer immunity to the newborn, especially through immunoglobulins (Ig) G. Melatonin is an immunomodulatory factor and there is evidence that it has a direct effect on IgG production. To evaluate the effects of melatonin treatment during pregnancy, litter size and offspring sex on colostrum quality, sixty pregnant Rasa Aragonesa ewes were divided into three groups: one group received a melatonin implant at the third month of pregnancy (3M, n = 13), another group at the fourth month (4M, n = 18) and the remaining ewes were not implanted (Control, C, n = 29). Immediately after lambing, a sample of colostrum was collected and IgG, crude protein and fat content analysed. Timing of melatonin implantation (p < .001), and offspring sex (p < .01) had a significant effect on IgG concentration. Colostrum of treated ewes had a higher mean (±SEM) IgG concentration than that of the control ewes (55.54 ± 3.09 and 49.50 ± 4.36 mg/ml, respectively; p < .05), mainly because the concentration in the 4M group was significantly (p < .001) higher than it was in the other groups. The relationship between lamb sex and IgG (p < .01) and %CP (p < .05) was evident in singletons (ewes with a male lamb: 54.57 ± 5.37 mg IgG/ml, 15.42 ± 0.82%CP; ewes with a female lamb: 34.66 ± 4.30 mg/ml, 13.18 ± 0.73%CP). The presence of a female in the litter was associated with significantly (p < .01) lower colostrum IgG levels (litters with at least one female: 49.33 ± 3.42 mg/ml; litters with no females: 58.24 ± 4.00 mg/ml). Among 4M ewes, female foetuses had significantly (p < .01) lower IgG levels whether they carried singletons or multiple lambs. In conclusion, treatment with melatonin implants at the fourth month of pregnancy resulted in a higher colostrum quality based on IgG concentration.

Disposable Amperometric Immunosensor for the Detection of Adulteration in Milk through Single or Multiplexed Determination of Bovine, Ovine, or Caprine Immunoglobulins G.

This paper reports the first immunoplatforms for the detection of adulteration in milk with milk or colostrum from other animals. The developed electrochemical bioplatforms allow the reliable determination of immunoglobulins G (IgGs) from cows, sheeps, or goats. They rely on sandwiching each animal species-specific IgGs with selective antibody pairs [unconjugated and conjugated with horseradish peroxidase (HRP)] onto magnetic microbeads (MBs) used as solid supports and amperometric transduction with the H2O2/hydroquinone (HQ) system at disposable electrodes. The immunoplatforms allow achieving limits of detection (LODs) of 0.74, 0.82, and 0.66 ng mL-1 for bovine, ovine, and caprine IgGs, respectively, which are lower than those obtained with conventional enzyme-linked immunosorbent assay (ELISA) methodologies and in 2-5 times shorter time. The bioplatforms were successfully applied to the determination of the individual content of the target IgGs in milk samples of different animals (cow, sheep, and goat) and type (colostrum, raw, and pasteurized), without matrix effect and after just a sample dilution. They were also applied to the detection of adulteration with milks from other animals at levels below than those required by the European legislation (1.0%, v/v). The possibility to detect milk adulteration with colostrum using a strategy based on the measurement of the total content of the three target IgGs in raw milks is also demonstrated. Multiplexing platforms were constructed to be used in routine surveillance of milk. They are able to provide in a single run and in just 30 min relevant information regarding the milk sample including its animal origin, the undergone heat treatment, and whether it was adulterated with milk or colostrum from other species.

Validation of the Microlab Salmonella for Detection of Salmonella O Group A-E in Selected Foods: AOAC Performance Tested MethodSM 052401.

BACKGROUND: Microlab Salmonella is an all-in-one solution for detection of Salmonella O Groups A-E that integrates, in a single disposable device, all elements to perform the analysis; a ready-to-use enrichment broth, a lateral flow immunochromatography test and a chemical agent for bacterial inactivation. Microlab Salmonella can be easily used on-site and does not require specific laboratory equipment or technical skills. The device is sealed during the analysis avoiding risks of contamination and is therefore safe to use in food production environments. OBJECTIVE: This report details the method validation study for raw ground beef, raw turkey (thermal processed, marinated), fresh cheese, deli ham and pasteurized liquid egg. METHODS: Matrix studies and inclusivity/exclusivity, product consistency/stability, and robustness testing were conducted to assess the method's performance. RESULTS: Inclusivity/exclusivity testing showed that the Microlab Salmonella method was able to detect Salmonella O Group A-E isolates while excluding the non-Salmonella strains. In the matrix studies the differences between the candidate and reference methods were not statistically significant. Minor variations in test parameters (enrichment time, time to read results, and enrichment temperature) did not affect the performance of the Microlab assay. There were no statistically significant differences between recently manufactured lots and those that were halfway through their expiration period or between those that were close to expiring. CONCLUSIONS: The reported data confirms that Microlab Salmonella is reliable for detecting Salmonella in raw ground beef, raw turkey (thermal processed, marinated), fresh cheese, pasteurized liquid egg and deli ham. HIGHLIGHTS: Microlab Salmonella is validated to detect Salmonella in select food matrixes. Microlab Salmonella integrates the steps of enrichment, detection, and inactivation in a single device.

Acute toxicity effects of pesticides on predatory snout mites (Trombidiformes: Bdellidae).

Predatory mites biologically control a range of arthropod crop pests and are often central to agricultural IPM strategies globally. Conflict between chemical and biological pest control has prompted increasing interest in selective pesticides with fewer off-target impacts on beneficial invertebrates, including predatory mites. However, the range of predatory mite species included in standardized pesticide toxicity assessments does not match the diversity of naturally occurring species contributing to biocontrol, with most testing carried out on species from the family Phytoseiidae (Mesostigmata). Here, we aim to bridge this knowledge gap by investigating the impacts of 22 agricultural pesticides on the predatory snout mite, Odontoscirus lapidaria (Kramer) (Trombidiformes: Bdellidae). Using internationally standardized testing methodologies, we identified several active ingredients with minimal impact on O. lapidaria mortality, including Bacillus thuringiensis, nuclear polyhedrosis virus, flonicamid, afidopyropen, chlorantraniliprole, and cyantraniliprole, which may therefore be good candidates for IPM strategies utilizing both chemical and biological control. Comparison of our findings with previous studies on Phytoseiid mites reveals important differences in responses to a number of chemicals between predatory mite families, including the miticides diafenthiuron and abamectin, highlighting the risk of making family-level generalizations from acute toxicity assessments. We also tested the impacts of several pesticides on a second Bdellidae species (Trombidiformes: Bdellidae) and found differences in the response to chlorpyrifos compared with O. lapidaria, further highlighting the taxon-specific nature of nontarget toxicity effects.

Acute toxicity effects of pesticides on beneficial organisms - Dispelling myths for a more sustainable use of chemicals in agricultural environments.

Agricultural practitioners, researchers and policymakers are increasingly advocating for integrated pest management (IPM) to reduce pesticide use while preserving crop productivity and profitability. Using selective pesticides, putatively designed to act on pests while minimising impacts on off-target organisms, is one such option - yet evidence of whether these chemicals control pests without adversely affecting natural enemies and other beneficial species (henceforth beneficials) remains scarce. At present, the selection of pesticides compatible with IPM often considers a single (or a limited number of) widely distributed beneficial species, without considering undesired effects on co-occurring beneficials. In this study, we conducted standardised laboratory bioassays to assess the acute toxicity effects of 20 chemicals on 15 beneficial species at multiple exposure timepoints, with the specific aims to: (1) identify common and diverging patterns in acute toxicity responses of tested beneficials; (2) determine if the effect of pesticides on beetles, wasps and mites is consistent across species within these groups; and (3) assess the impact of mortality assessment timepoints on International Organisation for Biological Control (IOBC) toxicity classifications. Our work demonstrates that in most cases, chemical toxicities cannot be generalised across a range of beneficial insects and mites providing biological control, a finding that was found even when comparing impacts among closely related species of beetles, wasps and mites. Additionally, we show that toxicity impacts increase with exposure length, pointing to limitations of IOBC protocols. This work challenges the notion that chemical toxicities can be adequately tested on a limited number of 'representative' species; instead, it highlights the need for careful consideration and testing on a range of regionally and seasonally relevant beneficial species.

Sensitive ELISA and lateral flow immunoassay for the detection of walnut traces in processed food and working surfaces.

Walnut represents one of the most allergenic nuts that can be found as a hidden allergen. In this study, sandwich ELISA and lateral flow immunoassay (LFIA), based on the determination of Jug r 1, were developed to detect walnut. Cross-reactivity was only found with Pecan nut among a panel of 88 food ingredients tested. ELISA and LFIA could detect 0.25 and 0.5 µg/g of walnut protein in complex food matrices spiked with walnut extract, respectively. Furthermore, walnut was detected in blended (chocolate) and incurred foods (ice cream and bread) added with ground walnut at levels of 0.5 and 1.5 µg protein/g by ELISA and LFIA, respectively. LFIA could also detect 0.1 µg of walnut protein in working surfaces. ELISA displayed acceptable precision and high recovery (71-97 %) and both tests were robust. This study shows that developed ELISA and LFIA are reliable tools to be applied in allergen control programs.

Quantitative determination of the okadaic acid toxins group by a colorimetric phosphatase inhibition assay: interlaboratory study.

An interlaboratory collaborative study to validate a colorimetric phosphatase inhibition assay for quantitative determination of the okadaic acid (OA) toxins group in molluscs, OkaTest, was conducted. Eight test materials, including mussels, scallops, clams, and cockles, were analyzed as blind duplicates. Blank samples and materials containing different OA toxin levels ranging from 98 to 275 microg/kg OA equivalents were included. The study was carried out by a total of 16 laboratories from 11 different countries. Values obtained for repeatability relative standard deviations (RSDr) ranged from 5.4 to 11.2% (mean 7.5%). Reproducibility RSD (RSD(R)) values were between 7.6 and 13.2% (mean 9.9%). The Horwitz ratio (HorRat) values ranged between 0.4 and 0.6. A recovery assay was also carried out using a sample spiked with OA. A mean recovery of 98.0% and an RSD of 14.5% were obtained. The results obtained in this validation study indicate that the colorimetric phosphatase inhibition assay, OkaTest, is suitable for quantitative determination of the OA toxins group. OkaTest could be used as a test that is complementary to the reference method for monitoring the OA toxins group.

[Clinical features of pericarditis in patients with SARS-CoV-2]. / Características clínicas de la pericarditis en pacientes con SARS-CoV-2.

Background: COVID-19 disease (coronavirus disease 2019) has multiple potentially fatal cardiovascular complications and pericarditis is one of them; however, if prompt treatment is given, fatal events associated to this complication decrease. Its frequency and presentation characteristics are unknown, which is why its early diagnosis is important. Objective: To know the frequency of pericarditis secondary to COVID-19 and its presentation characteristics. Material and methods: Cross-sectional study in patients with a diagnosis of pericarditis after COVID-19 disease (with a positive test). Symptoms, age, sex, comorbidities, and electrocardiogram (ECG) and transthoracic echocardiogram (TTE) results were obtained. Results: A total of 3364 patients positive for COVID-19 were registered, out of which 10 met criteria for pericarditis, which represented a frequency of 0.30%. The average age of the sample was 46.1 years and 60% predominated in the male gender with a 1.5:1 ratio. The most frequent clinical characteristics were the presence of retrosternal pain (90%), absence of comorbidity (50%), and absence of electrocardiographic changes (40%). Conclusions: Pericarditis has a low frequency in patients with COVID-19. The predominant clinical presentation is chest pain. Almost half will not have electrocardiographic changes, and half will not have comorbidities.

Introducción: la enfermedad COVID-19 (coronavirus disease 2019) tiene múltiples complicaciones cardiovasculares potencialmente mortales y la pericarditis es una de ellas; sin embargo, si se da un tratamiento oportuno disminuyen sus eventos fatales. Se desconoce su frecuencia y características de presentación, por lo que es importante su diagnóstico temprano. Objetivo: conocer la frecuencia de pericarditis secundaria a COVID-19 y sus características de presentación. Material y métodos: estudio transversal en pacientes con diagnóstico de pericarditis posterior a la enfermedad COVID-19 (con prueba positiva). Se obtuvo la sintomatología, edad, sexo, comorbilidades y resultados de electrocardiograma (ECG) y ecocardiograma transtorácico (ETT). Resultados: se registraron 3364 pacientes positivos a COVID-19, de los cuales 10 cumplieron con criterios para pericarditis, lo que representó una frecuencia del 0.30%. La edad promedio de la muestra fue de 46.1 años y predominó en el género masculino (60%) con una relación 1.5:1. Las características clínicas más frecuentes fueron la presencia de dolor retroesternal (90 %), ausencia de comorbilidad (50 %) y ausencia de cambios electrocardiográficos (40%). Conclusiones: la pericarditis tiene una frecuencia baja en pacientes con COVID-19. La presentación clínica predominante es el dolor retroesternal. Casi la mitad no tendrá cambios electrocardiográficos y la mitad no tendrá comorbilidades.

Systemic lupus erythematosus and systemic autoimmune connective tissue disorders behind recurrent diastolic heart failure.

Diastolic heart failure (DHF) remains unexplained in some patients with recurrent admissions after full investigation. A study was directed for screening SLE and systemic autoimmune connective tissue disorders in recurrent unexplained DHF patients admitted at a short-stay and intermediate care unit. It was found that systemic autoimmune conditions explained 11% from all of cases. Therapy also prevented new readmissions. Autoimmunity should be investigated in DHF.

Development of sandwich ELISA and lateral flow immunoassay to detect almond in processed food.

Almond (Prunus dulcis) represents a potential allergenic hazard that should be included in Allergen Control Plans. In this study, sandwich ELISA and lateral flow immunoassay (LFIA), using amandin (Pru du 6) as the target protein, were developed to detect almond in processed food and validated according to international guides. ELISA could detect 2 ng/mL and LFIA 30 ng/mL of pure amandin. No cross-reactivity was found on a panel of 50 food commodities with the exception of Pecan nut, Brazil nut and chestnut for which the cross-reactivity was lower than 0.02%. Furthermore, ELISA and LFIA were able to detect 0.12 and 0.70 ppm of almond protein in foods spiked with almond extract whereas 0.20 and 2.0 ppm could be detected in baked cookies incurred with almond, respectively. Both techniques could be applied for food manufacturers and control agencies for monitoring the presence of almond traces in food and working surfaces.

Ultrasensitive detection of soy traces by immunosensing of glycinin and ß-conglycinin at disposable electrochemical platforms.

This work reports the first electrochemical bioplatform for the determination of soy traces in food. The bioplatform involves sandwich-type immunoassays using specific antibodies for ß-conglycinin and glycinin, which are the main allergenic soy proteins, and carboxylic acid-modified magnetic microbeads. Amperometric detection at -0.20 V (vs. an Ag pseudo-reference electrode) was performed using single or dual screen-printed carbon electrodes and the H2O2/hydroquinone (HQ) system. The measured variation in the cathodic current was directly proportional to the concentration of target allergenic proteins. The developed bioplatforms exhibit a good selectivity and sensitivity providing limits of detection (LOD) values of 0.03 and 0.02 ng mL-1 for ß-conglycinin and glycinin, respectively. The determination of both proteins can be carried out in only 1.5 h. The electrochemical bioplatforms allow their accurate determinations (with results statistically comparable to those provided by ELISA methodologies) in raw cookie dough and baked cookies enriched with soy flour. The results obtained confirm, in a pioneering way with electrochemical biosensors, the possibility of discriminating samples incurred with as little as 0.0005 ppm of a food allergen in model cookie extracts.

Optimization and Validation of a New Microbial Inhibition Test for the Detection of Antimicrobial Residues in Living Animals Intended for Human Consumption.

Even though antibiotics are necessary in livestock production, they can be harmful not only due to their toxicity, but also in view of their contribution to the emergence of antimicrobial resistance. Screening tests based on microbial growth inhibition appeared to be useful tools to prevent its entry into the food chain. They have nevertheless been traditionally carried out post mortem, leading to great economical loss and harm to the environment in case a positive sample is found. Hence, the objective was to evaluate the use of a screening test as an ante mortem alternative for the detection of antibiotic residues in meat: thus, Explorer®-Blood test was optimized and validated. After adapting the procedure for matrix preparation, the assay parameters were assessed from 344 antibiotic-free blood serum samples. Limits of Detection (LoDs) were defined by spiking blood serum with several of the most common antimicrobials used in veterinary practice. LoDs were similar to those obtained for meat and were at or below the maximum residue limits set by EU legislation for muscle. Analyses of in vivo injected samples, previously characterized by LC-MS/MS, demonstrated the method's accuracy and proved that Explorer®-Blood can be considered a suitable alternative to conventional post mortem screening methods.

Validation of the Eclipse Farm 4G & COMET for Detection of Antibiotics in Raw Bovine Milk: AOAC Performance Tested MethodSM 022101.

BACKGROUND: The Eclipse Farm 4G test coupled to the COMET device allows for automatic and easy screening of a broad range of antimicrobials in raw bovine milk. All results obtained with this system are available in real-time through a smartphone application (App) and in the Test4all cloud platform. OBJECTIVE: The objective of this study was to validate Eclipse Farm 4G & COMET for the screening of antimicrobials in raw bovine milk according to AOAC Performance Tested MethodSM procedures. METHOD: The test is based on the inhibition of microbial growth of Geobacillus stearothermophilus in the presence of antimicrobials. When sample tubes are incubated in the absence of antibiotics, spores germinate and cells grow, changing the medium color from purple to yellow (negative result). The presence of antimicrobials in the milk sample hinders bacterial growth, preventing the tubes from changing color from purple to yellow (positive result). The COMET device automatically integrates all assay steps (incubation, time control, and results interpretation) and communications between the device, smartphone, and cloud. RESULTS: LODs and detection capabilities were confirmed to be at or below the European Union (EU) maximum residue limit (MRL) for most of the evaluated molecules representing the main families of antimicrobials (penicillins, cephalosporins, tetracyclines, sulfonamides, macrolides, and aminoglycosides). False-positive rates and the effect of potentially interfering substances showed sufficient selectivity/specificity. The Eclipse Farm 4G & COMET system was shown to be robust, consistent, and stable during shelf life. CONCLUSIONS: The Eclipse Farm 4G & COMET system has been shown to be suitable for screening antibiotics in raw bovine milk. HIGHLIGHTS: The assay is an automatic and easy system for the detection of a broad range of antibiotics in raw bovine milk consistent with EU MRLs and can provide results any time and anywhere through a smartphone App and a cloud platform.

Antibacterial Residue Excretion via Urine as an Indicator for Therapeutical Treatment Choice and Farm Waste Treatment.

Many of the infectious diseases that affect livestock have bacteria as etiological agents. Thus, therapy is based on antimicrobials that leave the animal's tissues mainly via urine, reaching the environment through slurry and waste water. Once there, antimicrobial residues may lead to antibacterial resistance as well as toxicity for plants, animals, or humans. Hence, the objective was to describe the rate of antimicrobial excretion in urine in order to select the most appropriate molecule while reducing harmful effects. Thus, 62 pigs were treated with sulfamethoxypyridazine, oxytetracycline, and enrofloxacin. Urine was collected through the withdrawal period and analysed via LC-MS/MS. Oxytetracycline had the slowest rate of degradation (a half-life time of 4.18 days) and the most extended elimination period in urine (over 2 months), followed by enrofloxacin (a half-life time of 1.48 days, total urine elimination in ca. 3 weeks) and sulfamethoxypyridazine (a half-life time of 0.49 days, total urine elimination in ca. 1 week). Bacterial sensitivity and recommendations for responsible use are limiting when selecting the treatment. Nevertheless, with similar effectiveness, sulfamethoxypyridazine would be the choice, as waste treatment would only need to be implemented for 1 week after treatment. Thus, more in-depth knowledge regarding antibacterial elimination would improve resource management, while protecting animals and consumers' health.

Influence of different extraction conditions on the detection of glycinin and ß-conglycinin in model processed foods by ELISA.

The presence of undeclared soy proteins in food can cause severe reactions in soy allergic individuals. The extraction of target proteins from processed foods is a crucial step in allergen detection by immunoassays, as only successfully extracted target proteins can be detected by the specific antibodies. The effectiveness was studied of different conditions (type of buffer, temperature and time of incubation) on the extraction of total protein, and concentration of glycinin and ß-conglycinin from different food matrices. The yields were determined using a soy protein isolate and three processed foods (sausage, bread and pâté) incurred with soy proteins. The yields were affected by the processing of analysed products and the composition and pH of the extraction buffers. Neutral and alkaline buffers (pH from 7.4 to 10.6) exhibited good protein extraction capacity and detectability of the specific target proteins. Denaturing additives and highly alkaline buffer (pH 12) extracted more crude protein but they were incompatible with the ELISA assay. Overall, the best results were obtained using phosphate (pH 7.4) and Tris/HCl (pH 8.5) buffers in the presence of 0.5 M NaCl. Crude protein yield of food extracts did not correlate with that of glycinin and ß-conglycinin, whereas a good relationship was found between the yields of the two proteins.

Is Blood a Good Indicator for Detecting Antimicrobials in Meat? Evidence for the Development of In Vivo Surveillance Methods.

The introduction of antimicrobial residues in the food chain has a significant impact on human health. An innovative solution to avoid their presence in meat is the adaptation of current control methods for use with in vivo matrixes. Thus, the aim was to obtain paired blood and muscle samples from pigs treated with some of the main antimicrobials currently used in veterinary medicine (oxytetracycline, sulfamethoxypyridazine, enrofloxacin, amoxicillin), and to compare their rate of depletion in both matrixes. Antimicrobial concentrations in paired samples of blood and muscle were determined by liquid chromatography with tandem mass spectrometry (LC-MS/MS) or high performance liquid chromatography with fluorescence detection (HPLC-FLD). A comparison between values obtained in muscle and blood showed a similar distribution in both matrixes for oxytetracycline; for sulfamethoxypyridazine, a similar decrease rate but a concentration three times higher in blood compared to muscle was found; for enrofloxacin, we found significant differences in the rate of depletion, with similar antimicrobial concentrations in both matrixes with values close to the maximum residue limit (MRL) and higher amounts in muscle for values that lay considerably over the MRL. Conversely, amoxicillin depletion was so rapid that its appearance in carcasses does not seem to pose a risk. Therefore, blood would be a feasible matrix for the development of new in vivo tests.