Dysregulated STAT3 signaling and T cell immunometabolic dysfunction define a targetable, high mortality subphenotype of critically ill children.

Sepsis is the leading cause of death of hospitalized children worldwide. Despite the established link between immune dysregulation and mortality in pediatric sepsis, it remains unclear which host immune factors contribute causally to adverse sepsis outcomes. Identifying modifiable pathobiology is an essential first step to successful translation of biologic insights into precision therapeutics. We designed a prospective, longitudinal cohort study of 88 critically ill pediatric patients with multiple organ dysfunction syndrome (MODS), including patients with and without sepsis, to define subphenotypes associated with targetable mechanisms of immune dysregulation. We first assessed plasma proteomic profiles and identified shared features of immune dysregulation in MODS patients with and without sepsis. We then employed consensus clustering to define three subphenotypes based on protein expression at disease onset and identified a strong association between subphenotype and clinical outcome. We next identified differences in immune cell frequency and activation state by MODS subphenotype and determined the association between hyperinflammatory pathway activation and cellular immunophenotype. Using single cell transcriptomics, we demonstrated STAT3 hyperactivation in lymphocytes from the sickest MODS subgroup and then identified an association between STAT3 hyperactivation and T cell immunometabolic dysregulation. Finally, we compared proteomics findings between patients with MODS and patients with inborn errors of immunity that amplify cytokine signaling pathways to further assess the impact of STAT3 hyperactivation in the most severe patients with MODS. Overall, these results identify a potentially pathologic and targetable role for STAT3 hyperactivation in a subset of pediatric patients with MODS who have high severity of illness and poor prognosis.

Influence of Native Saccharomyces cerevisiae Strains on Malvasia aromatica Wines.

BACKGROUND: In the search of tools to deal with climate change-related effects along with the aim of avoiding the loss of aromatic typicity in wine, two native yeasts strains of Saccharomyces cerevisiae (CLI 271 and CLI 889) were evaluated to determine their influence on white Malvasia aromatica wines aroma composition and sensory characteristics. METHODS: The strains were tested versus a commercial yeast strain (LSA). The fermentations were performed on grape must of the Malvasia aromatica variety previously macerated. Wine quality was studied by analysis of oenological parameters together with volatile aroma components using gas chromatography coupled to flame ionization detector (GC-FID) to quantify major volatiles compounds and headspace-solid phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME/GC-MS) to determine terpenoids and C13-norisoprenoids. Sensorial analysis was also realized by an experienced taster panel. RESULTS: Wines from locally-selected yeasts strains used had lower volatile acidity levels and higher concentration of aromatic compounds compared to the commercial strain ones. The yeast strain S. cerevisiae CLI 271 provided wines with a higher concentration of esters related to fruity attributes, especially isoamyl acetate. The tasting panel highlighted the strong floral character of wines from S. cerevisiae CLI 889 fermentation. CONCLUSIONS: The use of microorganisms well adapted to climatic conditions can be used to produce quality wines of the Malvasia aromatica variety.

Study of a First Approach to the Controlled Fermentation for Lambic Beer Production.

Non-Saccharomyces yeasts represent a great source of biodiversity for the production of new beer styles, since they can be used in different industrial areas, as pure culture starters, in co-fermentation with Saccharomyces, and in spontaneous fermentation (lambic and gueuze production, with the main contribution of Brettanomyces yeast). The fermentation process of lambic beer is characterized by different phases with a characteristic predominance of different microorganisms in each of them. As it is a spontaneous process, fermentation usually lasts from 10 months to 3 years. In this work, an attempt was made to perform a fermentation similar to the one that occurred in this process with lactic bacteria, Saccharomyces yeast and Brettanomyces yeast, but controlling their inoculation and therefore decreasing the time necessary for their action. For this purpose, after the first screening in 100 mL where eight Brettanomyces yeast strains from D.O. "Ribeira Sacra" (Galicia) were tested, one Brettanomyces bruxellensis strain was finally selected (B6) for fermentation in 1 L together with commercial strains of Saccharomyces cerevisiae S-04 yeast and Lactobacillus brevis lactic acid bacteria in different sequences. The combinations that showed the best fermentative capacity were tested in 14 L. Volatile compounds, lactic acid, acetic acid, colour, bitterness, residual sugars, ethanol, melatonin and antioxidant capacity were analysed at different maturation times of 1, 2, 6 and 12 months. Beers inoculated with Brettanomyces yeast independently of the other microorganisms showed pronounced aromas characteristic of the Brettanomyces yeast. Maturation after 12 months showed balanced beers with "Brett" aromas, as well as an increase in the antioxidant capacity of the beers.

Clinical Evolution and Risk Factors in Patients Infected during the First Wave of COVID-19: A Two-Year Longitudinal Study.

A limited number of longitudinal studies have examined the symptoms associated with long-COVID-19. We conducted an assessment of symptom onset, severity and patient recovery, and determined the percentage of patients who experienced reinfection up to 2 years after the initial onset of the disease. Our cohort comprises 377 patients (≥18 years) with laboratory-confirmed COVID-19 in a secondary hospital (Madrid, Spain), throughout March 3-16, 2020. Disease outcomes and clinical data were followed-up until August 12, 2022. We reviewed the evolution of the 253 patients who had survived as of April 2020 (67.1%). Nine died between April 2020 and August 2022. A multivariate regression analysis performed to detect the risk factors associated with long-COVID-19 revealed that the increased likelihood was associated with chronic obstructive lung disease (OR 14.35, 95% CI 1.89-109.09; p = 0.010), dyspnea (5.02, 1.02-24.75; p = 0.048), higher LDH (3.23, 1.34-7.52; p = 0.006), and lower D-dimer levels (0.164, 0.04-0.678; p = 0.012). Reinfected patients (n = 45) (47.8 years; 39.7-67.2) were younger than non-reinfected patients (64.1 years; 48.6-74.4)) (p < 0.001). Patients who received a combination of vaccines exhibited fewer symptoms (44.4%) compared to those who received a single type of vaccine (77.8%) (p = 0.048). Long-COVID-19 was detected in 27.05% (66/244) of patients. The early detection of risk factors helps predict the clinical course of patients with COVID-19. Middle-aged adults could be susceptible to reinfection, highlighting the importance of prevention and control measures regardless of vaccination status.

Non-Conventional Yeast: Behavior under Pure Culture, Sequential and Aeration Conditions in Beer Fermentation.

The use of wild yeasts, isolated from different environments, is becoming the most interesting option for the production of new beers. The objective of this study is to evaluate the potential of seven non-conventional yeast strains from five different species (Saccharomyces cerevisiae, Hanseniaspora guilliermondii, Metschnikowia pulcherrima, Torulaspora delbrueckii, and Zygosaccharomyces bailii) isolated from Madrid agriculture to produce type ale beer. Wild yeast strains were evaluated at laboratory and pilot plant scales under different fermentation conditions (pure, aerated, and sequential culture). Strain S. cerevisiae SafAle S-04 was used as a reference. Throughout the fermentation of beer, volatile compounds were determined by GC and residual sugars by HPLC, among other parameters. The yeast strains used for the fermentation in pure culture conditions were unable to ferment maltose and maltotriose (0.73-1.18% v/v of ethanol). The results of the study under aerated conditions showed varying levels of higher alcohol and ester concentrations. It should be noted that the strain CLI 1057 (S. cerevisiae) fermented maltose in the presence of oxygen (Kluyver effect). This strain also showed a high production of 4-vinyl guaiacol, making it suitable for producing beers with a phenolic profile. Finally, three strains (H. guilliermondii, Z. bailii, and T. delbrueckii) were evaluated in sequential culture together with commercial strain and found to improve the organoleptic characteristics of the brewed beer. These approaches offer the opportunity to add new product characteristics to the beers.

Impact of Non-Saccharomyces Wine Yeast Strains on Improving Healthy Characteristics and the Sensory Profile of Beer in Sequential Fermentation.

The use of non-Saccharomyces yeasts in brewing is a useful tool for developing new products to meet the growing consumer demand for innovative products. Non-Saccharomyces yeasts can be used both in single and in mixed fermentations with Saccharomyces cerevisiae, as they are able to improve the sensory profile of beers, and they can be used to obtain functional beers (with a low ethanol content and melatonin production). The aim of this study was to evaluate this capacity in eight non-Saccharomyces strains isolated from Madrid agriculture. For this purpose, single fermentations were carried out with non-Saccharomyces strains and sequential fermentations with non-Saccharomyces and the commercial strain SafAle S-04. The Wickerhamomyces anomalus strain CLI 1028 was selected in pure culture for brewing beer with a low ethanol content (1.25% (v/v)) for its fruity and phenolic flavours and the absence of wort flavours. The best-evaluated strains in sequential fermentation were CLI 3 (Hanseniaspora vineae) and CLI 457 (Metschnikowia pulcherrima), due to their fruity notes as well as their superior bitterness, body, and balance. Volatile compounds and melatonin production were analysed by GC and HPLC, respectively. The beers were sensory-analysed by a trained panel. The results of the study show the potential of non-Saccharomyces strains in the production of low-alcohol beers, and as a flavour enhancement in sequential fermentation.

Modeling the Impact of Climate Change on Cervid Chronic Wasting Disease in Semi-Arid South Texas.

Chronic wasting disease (CWD) is a spongiform encephalopathy disease caused by the transmission of infectious prion agents. CWD is a fatal disease that affects wild and farmed cervids in North America with few cases reported overseas. Social interaction of cervids, feeding practices by wildlife keepers and climate effects on the environmental carrying capacity all can affect CWD transmission in deer. Wildlife deer game hunting is economically important to the semi-arid South Texas region and is affected by climate change. In this paper, we model and investigate the effect of climate change on the spread of CWD using typical climate scenarios. We use a system of impulsive differential equations to depict the transmission of CWD between different age groups and gender of cervids. The carrying capacity and contact rates are assumed to depend on climate. Due to the polygamy of bucks, we use mating rates that depend on the number of bucks and does. We analyze the stability of the model and use simulations to study the effect of harvesting (culling) on eradicating the disease, given the climate of South Texas. We use typical climate change scenarios based on published data and our assumptions. For the climate indicator, we calculated and utilized the Standard Precipitation Evapotranspiration Index (SPEI). We found that climate change might hinder the efforts to reduce and effectively manage CWD as it becomes endemic to South Texas. The model shows the extinction of the deer population from this region is a likely outcome.

Temperatures Influence Susceptibility to Insecticides in Aedes aegypti and Aedes albopictus (Diptera: Culicidae) Mosquitoes.

Aedes aegypti and Aedes albopictus (Diptera: Culicidae) are vectors for several arboviruses, including dengue, Zika virus and chikungunya virus. The primary method of controlling these diseases is controlling the vector population, often with insecticides. Insecticide resistance may impact the success of these efforts. We tested the effect of variable temperature exposures on susceptibility to insecticides by exposing adult A.aegypti and A. albopictus to different temperatures and tested their susceptibility to insecticides. We hypothesized that adults maintained at high temperatures would show increased susceptibility to insecticides relative to lower temperatures. Colony mosquitoes were hatched, reared to adulthood and then maintained in three temperature regimes that reflect average seasonal temperatures in the Rio Grande Valley, TX. Susceptibility to permethrin and deltamethrin was assessed using the CDC bottle bioassay method. Overall Aedes albopictus had higher susceptibility to all insecticides than Aedes aegypti. Mosquitoes kept at different temperatures exhibited differential susceptibility to insecticides. Low temperature exposed mosquitoes had decreased susceptibility while high temperature conditions resulted in increased mortality. Our results suggest public health officials must consider temperature effects when controlling mosquitoes with insecticides.

Potential distribution of Amblyomma mixtum (Koch, 1844) in climate change scenarios in the Americas.

Amblyomma mixtum is a Neotropical generalist tick of medical and veterinary importance which is widely distributed from United States of America to Ecuador. The aim of this study was to evaluate changes in the geographic projections of the ecological niche models of A. mixtum in climate change scenarios in America. We constructed a database of published scientific publications, personal collections, personal communications, and online databases. Ecological niche modelling was performed with 15 Bioclimatic variables using kuenm in R and was projected to three time periods (Last Glacial Maximum, Current and 2050) for America. Our model indicated a wide distribution for A. mixtum, with higher probability of occurrence along the Gulf of Mexico and occurring in a lesser proportion in the Pacific states, Central America, and the northern part of South America. The areas of new invasion are located mainly on the border of Mexico with Guatemala and Belize, some regions of Central America and Colombia. We conclude that the ecological niche modelling are effective tools to infer the potential distribution of A. mixtum in America, in addition to helping to propose future measures of epidemiological control and surveillance in the new potential areas of invasion.

Diagnostic performance of CerTest and Panbio antigen rapid diagnostic tests to diagnose SARS-CoV-2 infection.

OBJECTIVES: Antigen rapid diagnostic tests (Ag-RDT) have been developed as reliable tools to control the SARS-CoV-2 pandemic. The objective of our study was to evaluate the diagnostic performance of two Ag-RDTs. METHODS: We evaluated CerTest SARS-CoV-2 Ag One Step Card Test and Panbio COVID-19 Ag Rapid Test Device Ag-RDTs. We included 320 nasopharyngeal samples: 150 PCR negative samples to assess the specificity and 170 PCR positive samples to evaluate the sensitivity. We also evaluated their sensitivity according to cycle threshold (Ct) values and the time from the onset of symptoms. Tests were compared using the McNemar's test and agreement was evaluated using the kappa score (k). RESULTS: Both Ag-RDTs showed a specificity of 100 %. Overall sensitivity was 53.5 % for CerTest and 60.0 % for Panbio. For samples with Ct≤ 25, sensitivity was 94.0 % for CerTest and 96.4 % for Panbio (p = 0.500). Regarding samples with Ct>25, sensitivity was 14.0 % for CerTest and 24.4 % for Panbio (p = 0.004). Sensitivity for samples within the first 5 days after the onset of symptoms were 84.8 % for CerTest and 91.3 % for Panbio (p = 0.250) and notably decreased for samples taken after the fifth day. Both Ag-RDTs showed an excellent agreement between them (agreement = 96.7 %, k = 0.920). Agreement with PCR was also excellent for high viral load samples (Ct<25) for CerTest (98.0 %, k = 0.954) and Panbio (98.8 %, k = 0.973). CONCLUSIONS: CerTest SARS-CoV-2 and Panbio COVID-19 Ag showed excellent performance and agreement results for samples with high viral loads (Ct ≤ 25) or samples taken within the first 5 days after the onset of symptoms.

Comparative evaluation of six immunoassays for the detection of antibodies against SARS-CoV-2.

OBJECTIVES: Serologic techniques can serve as a complement to diagnose SARS-CoV-2 infection. The objective of our study was to compare the diagnostic performance of six immunoassays to detect antibodies against SARS-CoV-2: three lateral flow immunoassays (LFAs), one ELISA and two chemiluminescence assays (CLIAs). METHODS: We evaluated three LFAs (Alltest, One Step and SeroFlash), one ELISA (Dia.Pro) and two CLIAs (Elecsys and COV2T). To assess the specificity, 60 pre-pandemic sera were used. To evaluate the sensitivity, we used 80 serum samples from patients with positive PCR for SARS-CoV-2. Agreement between techniques was evaluated using the kappa score (k). RESULTS: All immunoassays showed a specificity of 100 % except for SeroFlash (96.7 %). Overall sensitivity was 61.3 %, 73.8 %, 67.5 %, 85.9 %, 88.0 % and 92.0 % for Alltest, One Step, SeroFlash, Dia.Pro, Elecsys and COV2T, respectively. Sensitivity increased throughout the first two weeks from the onset of symptoms, reaching sensitivities over 85 % from 14 days for all LFAs, being One Step the most sensitive (97.6 %), followed by SeroFlash (95.1 %). Dia.Pro, Elecsys and COV2T showed sensitivities over 97 % from 14 days, being 100 % for COV2T. One Step showed the best agreement results among LFAs, showing excellent agreement with Dia.Pro (agreement = 94.2 %, k = 0.884), COV2T (99.1 %, k = 0.981) and Elecsys (97.3 %, k = 0.943). Dia.Pro, COV2T and Elecsys also showed excellent agreement between them. CONCLUSIONS: One Step, Dia.Pro, Elecsys and COV2T obtained the best diagnostic performance results. All these techniques showed a specificity of 100 % and sensitivities over 97 % from 14 days after the onset of symptoms, as well as excellent levels of agreement.

Panbio antigen rapid test is reliable to diagnose SARS-CoV-2 infection in the first 7 days after the onset of symptoms.

BACKGROUND: RT-qPCR is the current recommended laboratory method to diagnose SARS-CoV-2 acute infection, several factors such as requirement of special equipment, time consuming, high cost and skilled staff limit the use of these techniques. A more rapid and high-throughput method is essential. METHODS: We analyzed clinical data and nasopharyngeal samples, collected during September 2020, from patients attended at the emergency department of a secondary hospital and in two primary healthcare centers in Madrid. The performance of the Panbio™ COVID-19 AG Rapid Test Device for the detection of SARS-CoV-2 antigen was compared to RT-qPCR. RESULTS: 255 nasopharyngeal swabs, including 150 from the emergency department and 105 from primary helthcare centers, were tested. 184 patients were symptomatic (72.1 %). Amongst the 60 positive RT-qPCR samples, 40 were detected by the rapid antigen test, given an overall sensitivity of 73.3 %. All the samples detected positive with the rapid antigen test were also positive with RT-qPCR. The median cycle threshold was 23.28 (IQR 18.5-30.16). Patients with less than seven days onset of symptoms showed a higher viral load, and sensitivity for rapid antigen test (86.5 %), compared to those with more days (sensitivity of 53.8 %)(p < 0.004). CONCLUSIONS: The rapid antigen test evaluated in this study showed a high sensitivity and specificity in samples obtained during the first week of symptoms and with high viral loads. This assay seems to be an effective strategy for controlling the COVID-19 pandemic for the rapid identification and isolation of SARS-CoV-2 infected patients.

Alltest rapid lateral flow immunoassays is reliable in diagnosing SARS-CoV-2 infection from 14 days after symptom onset: A prospective single-center study.

OBJECTIVES: SARS-CoV-2 infection diagnosis is challenging in patients from 2 to 3 weeks after the onset of symptoms, due to the low positivity rate of the PCR. Serologic tests could be complementary to PCR in these situations. The aim of our study was to analyze the diagnostic performance of one serologic rapid test in COVID-19 patients. METHODS: We evaluated a lateral flow immunoassay (AllTest COVID-19 IgG/IgM) which detects IgG and IgM antibodies. We validated the serologic test using serum samples from 100 negative patients (group 1) and 90 patients with COVID-19 confirmed by PCR (group 2). Then, we prospectively evaluated the test in 61 patients with clinical diagnosis of pneumonia of unknown etiology that were negative for SARS-CoV-2 by PCR (group 3). RESULTS: All 100 patients from group 1 were negative for the serologic test (specificity = 100 %). Regarding group 2 (PCR-positive), the median time from their symptom onset until testing was 17 days. For these 90 group-2 patients, the test was positive for either IgM or IgG in 58 (overall sensitivity = 64.4 %), and in patients tested 14 days or more after the onset of symptoms, the sensitivity was 88.0 %. Regarding the 61 group-3 patients, median time after symptom onset was also 17 days, and the test was positive in 54 (88.5 % positivity). CONCLUSIONS: Our study shows that Alltest lateral flow immunoassay is reliable as a complement of PCR to diagnose SARS-CoV-2 infection after 14 days from the onset of symptoms and in patients with pneumonia and negative PCR for SARS-CoV-2.

Partial characterization of the voltage-gated sodium channel gene and molecular detection of permethrin resistance in Rhipicephalus annulatus (Say, 1821).

The cattle tick, Rhipicephalus annulatus (Say) is a vector of bovine babesiosis and responsible for direct and indirect losses to cattle producing areas located in temperate and subtropical dry regions. Resistance against pyrethroids has been reported for this species in Asia and Africa, but never before in North America. An outbreak strain, Rio Lado, collected close to the border between Mexico and the United States, in Maverick County, Texas, showed low level of resistance to permethrin, a pyrethroid pesticide. We used genomic material from different strains of cattle ticks collected within the Permanent Quarantine Zone (Rio Lado, Vega and Klein Grass strains) to partially characterize the coding gene of the voltage-gated sodium channel (Na-channel), target-site of pyrethroid pesticides, and search for putative mutations associated with resistance using quantitative PCR high resolution melt (HRM) analysis. The two amplified fragments, corresponding to domains II and III of the Na-channel, were 100 % identical to its ortholog in Rhipicephalus microplus (Canestrini). No nucleotide polymorphisms in the Na-channel gene were observed in the pyrethroid-resistant Rio Lado strain, when compared to the susceptible strains Klein Grass and Vega. This study reports the first case of pyrethroid resistance in R. annulatus collected in the United States. Also, we provide new genomic data for this species of tick that allows for the development of a new method to screen for mutations associated with pyrethroid resistance.

Eficacia y optimización de la citometría de flujo en el cribado universal de la infección del tracto urinario / Efficacy and optimisation of flow cytometry in the universal screening of urinary tract infection

Introducción: La citometría de flujo ha mostrado en los últimos años su utilidad en el cribado de infección urinaria y su integración en los Servicios de Microbiología podría evitar la siembra de hasta el 60% de las muestras del área. El propósito de este trabajo es valorar su utilidad en el cribado universal y la rentabilidad diagnóstica en diferentes subpoblaciones. Material y métodos: Se analizaron 1.338 muestras mediante citometría de flujo (Sysmex UF-1000i) y urocultivo en agar CPS. Se informaron como positivos los cultivos con 1 o 2 uropatógenos y recuentos superiores a 105 UFC/ml y en poblaciones especiales, recuentos inferiores con un solo uropatógeno. Resultados: Utilizando un punto de corte de>17,1 bacterias/μl o>29,5 leucocitos/μl se consiguió una sensibilidad del 95,15% y un porcentaje de cribado del 32,14%. De los once falsos negativos, seis presentaban recuentos bajos y en uno se cultivó Candida glabrata. En el estudio por subgrupos se encontraron variaciones estadísticamente significativas respecto al género y la procedencia. El punto de corte en los varones fue inferior con respecto a las mujeres. Sin embargo, se mantuvo estable en las muestras de Atención Primaria, reduciéndose sustancialmente en las hospitalarias. El valor predictivo negativo permaneció siempre por encima del 95%. Conclusión: El sistema de citometría de flujo puede evitar la siembra del 32% de las muestras aplicando criterios estrictos de positividad. En nuestro hospital, habría evitado la siembra de 13.705 orinas en 2016, y podría aumentar combinando los puntos de corte con la procedencia y el género de las muestras

Introduction: Flow cytometry has shown to be useful for ruling out urinary tract infection over the last few years. Its integration into the Microbiology Laboratories could avoid the urine culture of 60% of the samples. The aim of this study is to evaluate the usefulness of flow cytometry in the universal screening, as well as to improve its efficacy by using specific cut-off points in different groups. Material and methods: A total of 1338 urine samples were analysed by flow cytometry (Sysmex UF-1000i), as well as a urine culture in CPS agar. Cultures with one or two pathogens and more than 10,000 CFU/ml, and special cases with less counts but just one pathogen, were considered as positive. Results: A cut-off of >17.1 bacteria/μl or >29.5 leucocytes/μl resulted with a sensitivity of 95.15% and a screening yield of 32.14%. Eleven false negative were obtained, but six of them showed low counts, and another was due to Candida glabrata. On the other hand, statistically significant variations were found as regards gender and origin of the patients. The cut-off of male samples was lower than female ones. However, it remained stable in the samples from Primary Care, and it decreased notably in those from the hospital. The negative predictive value always remained over 95%. Conclusion: Automated flow cytometry can avoid the culture of 32% of samples, even after applying tight positive criteria. In the study hospital, it would have avoided the culture of 13,705 urine samples in the year 2016. These results could improve by combining cut-off points, gender, and origin of patients

Screening Sticky Cards as a Simple Method for Improving Efficiency of Diaphorina citri (Hemiptera: Liviidae) Monitoring and Reducing Nontarget Organisms.

Management of Diaphorina citri Kuwayama (Hemiptera: Liviidae) populations is one of the major strategies for reducing the spread and incidence of huanglongbing (HLB). HLB is putatively caused by Candidatus Liberibacter spp. (Rhizobiales: Phyllopbacteriaceae) that are transmitted to citrus by psyllid vectors. Diaphorina citri population monitoring is done to detect its presence and inform on management decisions. Various methods are used for detecting and estimating D. citri densities but trapping with yellow or lime-green sticky cards has proven to be the most effective method. These sticky cards rely on the color preference of adult D. citri, but many flying organisms are attracted to the same color spectrum as psyllids. Hence, in field situations, sticky traps are hampered by large numbers of bycatches of nontarget organisms and debris. Here, we described a method using a mesh laid on the surface of traps as a sift to catch mainly psyllids, while reducing bycatches. By filtering D. citri through this mesh, they can be counted more rapidly and accurately. Although mesh-covered traps captured 5-15% less D. citri relative to uncovered ACP traps, both types of traps statistically agreed on D. citri detection and population densities. The effectiveness of mesh-covered traps did not vary with season. In addition, mesh-covered traps eliminated >90% of nontarget organisms and allowed for quicker enumeration of D. citri. We expect this method will become an important component of redesigning integrated pest management programs in citrus groves by reducing unintended impacts of beneficial arthropods during large scale D. citri monitoring.

High-resolution melt (HRM) analysis for detection of SNPs associated with pyrethroid resistance in the southern cattle fever tick, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

The southern cattle fever tick, Rhipicephalus (Boophilus) microplus, is the most economically important ectoparasite of cattle worldwide. A limitation for sustainable control and eradication is the emergence of acaricide resistance among tick populations. Molecular diagnostic tools offer the opportunity to detect resistance rapidly, which can be complemented with confirmatory bioassays with larvae and adult ticks that are more resource and time consuming to generate. Synthetic pyrethroid resistance is one of the most prevalent and well-studied forms of resistance in arthropods, being linked with target site alterations in the sodium ion channel gene. Here, we report research on a novel molecular method to detect mutations in the para-sodium channel gene of R. microplus associated with acaricide resistance that is based on quantitative PCR high-resolution melt (HRM) analysis. Genomic DNA fragments of domains II and III of the para-sodium channel gene were amplified by real-time PCR in the presence of EVA®Green dye to test resistant and susceptible reference ticks from the U.S., Brazil, and Mexico. Larval packet tests with discriminating doses and a modified lethal time analysis were performed to confirm resistance to permethrin, cypermethrin, deltamethrin, and flumethrin in laboratory strains. Tick specimens collected from cattle that were inspected at the United States Port-of-Entry at the Texas-Mexico border were also genotyped. Previously described mutations associated with pyrethroid resistance (T170C, C190A, G184C, and T2134A) were successfully detected by qPCR-HRM in different genotypes and confirmed by sequencing. A novel non-synonymous SNP located at domain III (C2136A) and the G215T mutation in domain II, previously described only in Asian R. microplus and R. australis, were also detected with the HRM and confirmed by sequencing. This technique could be adapted for high-throughput screening, detection, and discovery of allele-specific mutations in cattle tick outbreak populations to inform eradication strategies in the USA. This knowledge could also be applied to integrated control programs in other parts of the world where R. microplus is endemic and where similar SNPs have been identified associated with pyrethroid resistance. This study highlights the existence of several mutations in the para-sodium channel gene in different combinations in field populations of R. microplus from Mexico.

Quantification of Wine Mixtures with an Electronic Nose and a Human Panel.

In this work, an electronic nose and a human panel were used for the quantification of wines formed by binary mixtures of four white grape varieties and two varieties of red wines at different percentages (from 0 to 100% in 10% steps for the electronic nose and from 0 to 100% in 25% steps for the human panel). The wines were prepared using the traditional method with commercial yeasts. Both techniques were able to quantify the mixtures tested, but it is important to note that the technology of the electronic nose is faster, simpler, and more objective than the human panel. In addition, better results of quantification were also obtained using the electronic nose.

Effect of seasonal climate fluctuations on the evolution of glycoconjugates during the ripening period of grapevine cv. Muscat à petits grains blancs berries.

BACKGROUND: Temperature changes and rainfall fluctuations affect the winemaking industry by altering vine phenology, crop yield, must composition and wine quality. Here, the evolution of the varietal aroma potential during ripening over three consecutive seasons has been analyzed considering climate conditions. The effect of season, ripening date and climatic conditions on aroma precursor accumulation in grapes was studied by multivariate analysis and discussed in regard to the potential aromatic quality of the wine. RESULTS: In contrast to musts obtained from grapes harvested in warm and dry seasons (2014 and 2015), musts obtained from grapes harvested in the coolest and most humid year (2013) showed a lower pH, higher total acidity, higher varietal aroma potential index (IPAv), and higher aromatic potential of linalool, geraniol and nerol (typical of the Muscat flavor). CONCLUSION: Considering the three seasons, grapes synthesized and accumulated more precursors when milder temperatures and late rains throughout veraison were recorded. However, we observed a high fluctuation of aroma precursors during the ripening period, so choosing the correct harvesting date (according to grape aroma potential) may overcome some of the negative effects linked to climate conditions. © 2017 Society of Chemical Industry.

Polysaccharides and Oligosaccharides Produced on Malvar Wines Elaborated with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts from D.O. "Vinos de Madrid".

Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. Both fractions from different white wines were separated by high-resolution size-exclusion chromatography. Glycosyl composition and wine polysaccharide linkages were determined by GC-EI-MS chromatography. Molar-mass distributions were determined by SEC-MALLS, and intrinsic viscosity was determined by differential viscometer. Yeast species and type of inoculation have a significant impact on wine carbohydrate composition and structure. Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. Moreover, we reported structural differences between mannoproteins released by T. delbrueckii CLI 918 and those released by S. cerevisiae CLI 889. These findings help to provide important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species.