Daily Samples Research Articles

Assessing daily patterns in stimulant use during the COVID-19 pandemic in Melbourne, Australia using wastewater analysis

During the COVID-19 pandemic, one of Australia’s biggest cities, Melbourne, experienced three major isolation (“lockdown”) periods in 2020 (160 days) and in 2021 (111 days) which makes it one of the most locked down cities world-wide. This study assessed how the pandemic affected temporal trends in methamphetamine, MDMA and cocaine consumption using wastewater-based epidemiology. Daily samples were collected for most of 2020 and 2021 (n = 660 days). Concentrations were measured using direct-injection LC-MS/MS and back-calculated to consumption estimates. Results indicate that methamphetamine use was increasing before the first lockdown and decreased after the end of the first lockdown in 2020. Methamphetamine trends appeared to have remained steady throughout the second lockdown period before increasing steeply after it ended. For most of 2020, cocaine use remained steady, with an increase after the second lockdown. MDMA use decreased after the start of the first lockdown and remained steady throughout most of 2020 and 2021. In comparison to 2020, trends in 2021 were less variable and stimulant use did not appear to be as associated with COVID-19 restrictions. Overall, this study was able to show the impact of lockdown periods and the related social restrictions on illicit stimulant use. Environmental ImplicationIllicit drugs are hazardous chemicals, of concern both to humans and the environment. While studies have been undertaken to understand their temporal trends, this work utilizes wastewater-based epidemiology and daily sampling to provide a comprehensive understanding of the impact of the COVID-19 pandemic on the use of methamphetamine, MDMA and cocaine on one of the most locked-down cities in the world. Understanding the consequences of this significant intervention on illicit drug use could provide valuable insights into its potential environmental impact.

Spatiotemporal distribution of oxidative potential in PM2.5 and its key components across six Chinese cities

Increasing evidence has supported that oxidative potential (OP) serves as a crucial indicator of health risk of exposure to PM2.5 over mass concentration. However, there is a lack of comparative studies across multiple cities, particularly on a fine temporal scale. In this study, we aim to investigate daily variation of ambient PM2.5 OP through simultaneous samplings in six Chinese cities for one year. Results showed that more than 60 % of the sampling days exhibited non-zero ranking difference between volume-normalized oxidative potential (OPv) and mass concentration among the six cities. Key components contributing to OPv inculde Mn, NO3-, and K+, followed by Ca2+, Al, SO42-, Cl-, Fe, and NH4+. Based on these chemical components, we developed a stepwise multivariable linear regression model (R2: 0.71) for OPv prediction. The performance of the model is comparable to both species- and sources-based ones in the literature. These findings suggest that a relatively lower daily-averaged mass concentration of PM2.5 does not necessarily indicate a lower oxidative risk. Future studies and policy developments on health benefits should also consider OPv rather than mass concentration alone. Priority could be given to sources/species that contribute significantly to oxidative potential of ambient PM2.5. SynopsisThis study highlights inclusion of oxidative potential as a complementary metric for air pollution assessment and control.

Impacts of anthropogenic habitat modification on mammalian diversity in the Mau Forest Complex, Kenya

Habitat destruction is a major driver of biodiversity loss, especially in the tropics. We investigated changes in the diversity and occupancy of forest-dwelling mammals in response to habitat loss, alteration, and degradation at the Mau Forest Complex (MFC), southwestern Kenya. Using a systematic camera trapping grid, we estimated patterns of mammalian diversity, occupancy of small carnivores, and detection rates of functional mammalian groups across three habitat types (primary forests, secondary forests, and cultivated habitats). Between March 2019 and October 2021, 246 camera traps were deployed across 181 locations representing 5509 sampling days, resulting in 47,345 images of 52 mammalian species in nine mammalian orders. Alpha diversity differed among treatments, with primary forests having the highest diversity, and cultivated habitats the lowest, though results were not statistically significant. Beta diversity differed significantly among treatments, indicating differences in community composition as well as species richness. Overall occupancy for small carnivores was highest in primary forests and lowest in cultivated habitats, with some species specializing in primary forests while others were able to occupy more degraded habitats. Among functional groups, detection rates only differed significantly for large rodents, primates, and ungulates. Our results indicate that although functional groups and species may respond differently to habitat loss and alteration at the MFC, overall species diversity of specialist mammals was highest in the less-disturbed primary forests. Specialist, forest-dependent species are likely most affected by human activities, making them especially vulnerable to habitat loss at the MFC.

P-105 Gender affirming hormone therapy affects the spermatogonial cell compartment in transwomen

Abstract Study question What are the specific changes under gender affirming hormone therapy (GAHT) in the spermatogonial cell compartment in testicular tissues of transwomen? Summary answer GAHT has a negative effect on the most undifferentiated PIWIL4+ spermatogonia potentially leading to reduced chances for fertility preservation in transwomen. What is known already Transwomen take GAHT to achieve transition between the gender assigned at birth and their gender identity. GAHT leads to a reduction of the most advanced germ cells and a reduction of MAGEA4 positive spermatogonia (pan spermatogonial marker). Single-cell RNA sequencing studies identified spermatogonial subpopulations based on the expression of the marker genes PIWIL4, FGFR3, NANOS3, GFRA1, KIT, UTF1. PIWIL4+ cells are considered the origin of germ cell differentiation (state 0). State 1 and 2 spermatogonia are characterized by expression of GFRA1 and KIT, respectively. Study design, size, duration On the day of gender affirming surgery (GAS) testicular tissues and blood samples were collected of 25 age matched trans women (mean 28.1 yr), who underwent comparable GAHT regimens (10 or 12.5 mg cyproterone acetate and estrogens) between 2013 and 2018. 8 adult cis men (mean 34.5 yr) with complete spermatogenesis served as controls. Age, start and type of GAHT were assessed using a questionnaire. This study was approved by the local ethics committee. Participants/materials, setting, methods We performed immunohistochemical stainings to evaluate the expression of spermatogonial markers (MAGEA4, UTF1, PIWIL4, FGFR3, NANOS3, GFRA1) in Bouin’s-fixed testicular tissue sections. For each marker the number of positive cells per round tubular cross-section was determined in 25 round tubules. The spermatogonial proliferation rate was evaluated using immunofluorescence co-stainings for GFRA1/MIKI67/MAGEA4 and KIT/MKI67/MAGEA4. We scored 400 MAGEA4+ cells per individual and determined the proportion of GFRA1+/KIT+. MKI67+ cells among the GFRA1+/KIT+ were evaluated. Main results and the role of chance The number of MAGEA4+ spermatogonia and the proportion of PIWIL4+ spermatogonia within the MAGEA4+ spermatogonia were reduced in transwomen compared to controls (p < 0.001). In contrast, the relative proportion of UTF1+, FGFR3+ and NANOS3+ spermatogonia was comparable. The number of spermatogonia per tubule correlated negatively with the age at start of GAHT (p < 0.01) and the age on the day of GAS (p < 0.001). The treatment duration correlated negatively with the proportion of PIWIL4+ (p < 0.01) and NANOS3+ spermatogonia (p < 0.05). The immunofluorescence stainings in transwomen revealed a similar ratio of proliferating KI67+ spermatogonia, both within the GFRA1+ and KIT+ cells, compared to controls. These results show the negative effects of the GAHT itself on the population of spermatogonia as well as the individual susceptibility depending on age-related factors. These results have the potential to better define the chances for fertility preservation under GAHT. Limitations, reasons for caution Transwomen are a very heterogeneous patient group varying in age, different GAHT, different treatment durations, intake of co-medications and possible pre-existing medical conditions. Hence, testicular size and histological appearance of transwomen’s testes are heterogeneous. Wider implications of the findings Transwomen, who wish to have own biological children, have to be counseled with regard to fertility preservation, best before initiation of GAHT, in order to avoid adverse effects on spermatogonia. Individual counselling has to take place with regard to fertility preservation. Trial registration number 2012-555-f-S

Validating a microphysical prognostic stratospheric aerosol implementation in E3SMv2 using observations after the Mount Pinatubo eruption

Abstract. This paper describes the addition of a stratospheric prognostic aerosol (SPA) capability – developed with the goal of accurately simulating sulfate aerosol formation and evolution in the stratosphere – in the Department of Energy (DOE) Energy Exascale Earth System Model, version 2 (E3SMv2). The implementation includes changes to the four-mode Modal Aerosol Module microphysics in the stratosphere to allow for larger particle growth and more accurate stratospheric aerosol lifetime following the Pinatubo eruption. E3SMv2-SPA reasonably reproduces stratospheric aerosol lifetime, burden, aerosol optical depth, and top-of-atmosphere flux when compared to remote sensing observations. E3SMv2-SPA also has close agreement with the interactive chemistry–climate model CESM2-WACCM (Community Earth System Model version 2–Whole Atmosphere Community Climate Model) – which has a more complete chemical treatment – and the observationally constrained, prescribed volcanic aerosol treatment in E3SMv2. Global stratospheric aerosol size distributions identify the nucleation and growth of sulfate aerosol from volcanically injected SO2 from both major and minor volcanic eruptions from 1991 to 1993. The modeled aerosol effective radius is consistently lower than satellite and in situ measurements (max differences of ∼ 30 %). Comparisons with in situ size distribution samples indicate that this simulated underestimation in both E3SMv2-SPA and CESM2-WACCM is due to overly small accumulation and coarse-mode aerosols 6–18 months post-eruption, with E3SMv2-SPA simulating ∼ 50 % of the coarse-mode geometric mean diameters of observations 11 months post-eruption. Effective radii from the models and observations are used to calculate offline scattering and absorption efficiencies to explore the implications of smaller simulated aerosol size for the Pinatubo climate impacts. Scattering efficiencies at wavelengths of peak solar irradiance (∼ 0.5 µm) are 10 %–80 % higher for daily samples in models relative to observations through 1993, suggesting higher diffuse radiation at the surface and a larger cooling effect in the models due to the smaller simulated aerosol; absorption efficiencies at the peak wavelengths of outgoing terrestrial radiation (∼ 10 µm) are 15 %–40 % lower for daily samples in models relative to observations, suggesting an underestimation in stratospheric heating in the models due to the smaller simulated aerosol. These potential biases are based on aerosol size alone and do not take into account differences in the aerosol number. The overall agreement of E3SMv2-SPA with observations and its similar performance to the well-validated CESM2-WACCM makes E3SMv2-SPA a viable alternative to simulating climate impacts from stratospheric sulfate aerosols.

Insights into the uptake, translocation, and accumulation dynamics of cyantraniliprole and thiamethoxam seed coating pesticides in maize plants.

Seed coating with pesticides is used extensively for the protection of both seeds and plants against pests. In this study, the uptake and transport of seed-coating pesticides (insecticides), including cyantraniliprole (CYN) and thiamethoxam (THX), were investigated. The translocation of these pesticides from the soil to the plant and their accumulation in different plant parts were also calculated. After sowing the seeds with seed coating pesticides, soil and plant samples were taken across the study area. These samples were extracted and analyzed in liquid chromatography with tandem mass spectrometry (LC-MS/MS). CYN and THX were used in maize plants for the first time to observe soil degradation kinetics, and CYN showed a higher half-life than THX in soil. Both pesticides have been taken up by the corn maize plant and transferred and accumulated to the upper parts of the plant. Although the THX concentration was between 2.240 and 0.003mg/kg in the root, between 3.360 and 0.085mg/kg in the stem, it was between 0.277 and 3.980mg/kg in the leaf, whereas CYN was detected at higher concentrations. The concentration of CYN was 1.472mg/ kg and 0.079mg/kg in the roots and stems of the maize plant, respectively. However, the bioconcentration factor (BCF) indicates the soil-to-plant accumulation of CYN from 28 to 34.6 and that of 12.5 to 4567.1 for THX on different sampling days. The translocation factor (TFstem) represents the ratio of pesticides absorbed from the stem and transported to the roots. For CYN, TFstem ranges from 3.6 to 20.5, while for THX, it varies between 1.5 and 26.8, indicating a higher translocation rate for THX. The ratio of leaf to root concentration are 3.6 to 20.5 for CYN and 1.8 to 87.7 for THX, demonstrating effective translocation for both pesticides. The TF values for both pesticides are above 1, signifying successful root-to-stem-to-leaf movement. Notably, THX exhibits a notably higher transport rate compared to CYN.

Influence of Controlled Condition Ice Encasement of Creeping Bentgrass and Annual Bluegrass on Plant Recovery, Gas Evolution, and Metabolites

Two cool-season putting green turfgrass species, annual bluegrass and creeping bentgrass, are differential in ice encasement tolerance. Physiological mechanisms associated with creeping bentgrass ice encasement tolerance and annual bluegrass susceptibility are not understood. The objectives were to evaluate oxygen, ethylene, and CO2 content within the upper soil space of the plants while frozen and immediately after ice melt after 0, 5, 10, 20, and 28 days of ice encasement (2.54 cm of ice) in growth chamber conditions. Following ice melt, plant samples were separated into leaf, crown, and root tissues and used to evaluate carbohydrate and amino acid content. Annual bluegrass exhibited higher damage (slower recovery rates) on most sampling days compared with creeping bentgrass. The organs that were most damaged and exhibited a differential principal component analysis snapshot, were the leaf and crown tissues. Creeping bentgrass may preserve leaf and crown tissues for postwinter recovery whereas significant metabolic changes occur in annual bluegrass leaves and crowns. Creeping bentgrass retained total amino acids in leaves following ice encasement whereas total leaf amino acid levels declined in annual bluegrass. Specific carbohydrates and amino acids such as the ability to maintain high levels of fructose, asparagine, and proline may be important indicators of the tolerance to ice encasement stress. On the basis of more prominent carbohydrate and amino acid loss in leaves and crowns and higher levels of CO2 evolution, annual bluegrass may exhibit a higher metabolism and/or tissue damage during ice encasement compared with creeping bentgrass, which could reduce spring recuperative potential.

Chronic Heat Stress Induces Oxidative Stress and Induces Inflammatory Injury in Broiler Spleen via TLRs/MyD88/NF-κB Signaling Pathway in Broilers.

The spleen is the largest peripheral immune organ of the organism, accounting for 25% of the total lymphoid tissue of the body. During HS, the spleen is damaged due to the elevated environment, which seriously affects life performance and broilers' health. This study aimed to investigate the mechanism of chronic HS damage to broiler spleen tissues. The broilers were typically raised until they reached 21 days of age, after which they were arbitrarily allocated into two groups: an HS group and a cntrol group. The HS group was subjected to a temperature of 35 °C for 10 h each day, starting at 21 days of age. At 35 and 42 days of age, spleen and serum samples were obtained from the broilers. The results showed that after HS, a significant decrease in productive performance was observed at 42 days of age (p < 0.01), and the spleen index, and bursa index were significantly decreased (p < 0.01). T-AOC of the organism was significantly decreased (p < 0.05), GSH-PX, SOD, and CAT antioxidant factors were significantly decreased (p < 0.01), and MDA was significantly elevated (p < 0.01). HS also led to a significant increase in cytokines IL-6, TNF-α, and INF-γ and a significant decrease in IL-4 in the spleen. The histopathologic results showed that the spleen's red-white medulla was poorly demarcated. The cells were sparsely arranged after HS. After HS, the expression of TLRs, MYD88, and NF-κB genes increased significantly. The expression of HSP70 increased significantly, suggesting that HS may induces an inflammatory response in broiler spleens through this signaling pathway, which may cause pathological damage to broiler spleens, leading to a decrease in immune function and progressively aggravating HS-induced damage with the prolongation of HS.

Continuous daily sampling of airborne eDNA detects all vertebrate species identified by camera traps

AbstractOngoing pressures on global biodiversity require conservation action that is not possible without effective biomonitoring. Terrestrial vertebrate surveys are commonly performed using camera traps, a time‐intensive method known to miss many small or arboreal species and birds. Recent advances have shown airborne eDNA to be a potentially suitable technique to more effectively monitor vertebrate communities in a time‐ and cost‐effective manner. Here, we test whether commercially available air samplers that collect air particles 24/7 during a 1‐week period can be used to detect the presence of vertebrates through airborne eDNA. The results are compared to camera trap records at three locations with differing habitats in the Netherlands. Simultaneous sampling with three different air samplers for 3 weeks resulted in detection of 154 vertebrate taxa, of which the majority were birds or mammals (113 and 33 species, respectively), along with four fish and four amphibian species. All species observed using camera traps were also retrieved via airborne eDNA, although not on every day of sampling. The Burkard spore trap, used routinely for pollen monitoring, showed the highest number of vertebrate species, and only in three samples when a mammal species was detected using a camera trap it remained undetected via eDNA. We also detected unique species at the three locations using airborne eDNA, indicative of the habitat in which they were living. However, we also detected species that we could not account for. The multitude of species found using airborne eDNA compared to camera traps indicate the sensitivity of the method; however, subsequent studies should prioritize validation of these findings through alternative biomonitoring approaches.

Superovulating cattle with corifollitropin-alpha, a long-acting recombinant human FSH (rhFSH): Dose-response, half-life, effects on the ovaries, and embryo outcomes

The aim of this study was to evaluate the superestimulatory and superovulatory responses of cattle treated with corifollitropin-alpha, a long-acting human recombinant FSH (rhFSH). In the first and second experiments, we used Nelore (Bos indicus) heifers previously submitted to follicular wave suppression by active immunization against GnRH. In Experiment 1 (a dose-response study), heifers (n = 20) were randomly allocated into five groups, which received placebo (saline) or a single sc dose of 7.5, 15.0, 22.5 or 30.0 μg rhFSH. The heifers were subjected to daily ovarian scan and blood sampling during 11 days. We observed group, time, and group x time effects (P<0.0001) for both average follicle size and circulating FSH concentrations, with a strong correlation (R = 0.82, P<0.0001) between the area under curve (AUC) for both parameters. The peak concentration of FSH 24h after treatment and average follicle size at all timepoints, however, were similar (P>0.05) between groups 22.5 and 30.0 μg. In Experiment 2, heifers (n = 18) were allocated into three groups, which received (0h) either placebo (control), 25 μg rhFSH or 130 mg pFSH (Folltropin). There was no difference (P>0.05) in average follicle size at any moment, as well as in intrafollicular E2 at 120h or in plasma P4 seven days later between groups rhFSH and pFSH. In Experiment 3, cycling Nelore heifers (n = 20) were subjected to a wave synchronization protocol and superovulated (day 0) using a standard pFSH protocol (120 mg split in eight decreasing im doses) or with a single sc injection of 20 μg rhFSH. The number of follicles >7 mm on day 4 did not differ (P=0.4370). Heifers receiving rhFSH had greater average follicle size on day 4 (P=0.0005), ovulation rate (P<0.0001), and number of CL (P=0.0155), as well as a trend towards a greater number of ova (P=0.07) and viable embryos (P=0.0590). In Experiment 4, superovulation was induced with a single sc injection of 25 μg rhFSH in Girolando and Nelore cows and heifers (n = 20). None of the embryo yield endpoints differed between the two breeds (P>0.05). In conclusion, cattle superstimulation and superovulation can be successfully induced with a single dose of a long-acting rhFSH (corifollitropin-alpha).

Wastewater-based epidemiology to monitor 68 NPS/conventional drug use in Taipei metropolitan area in Taiwan during and after COVID-19 pandemic

Amidst far-reaching COVID-19 effects and social constraints, this study leveraged wastewater-based epidemiology to track 38 conventional drugs and 30 new psychoactive substances (NPS) in northern Taiwan. Analyzing daily samples from four Taipei wastewater plants between September 2021 and January 2024—encompassing club reopenings, holidays, Lunar New Year, an outbreak, and regular periods—thirty-one drugs were detected, including 5 NPS. Tramadol, zolpidem tartrate, CMA, and MDPV were newly detected in Taiwanese sewage with frequency of 1.4 %− 89.0 %. Conventional drug use typically increased post-pandemic, aside from benzodiazepines and methadone. Methamphetamine showed 100 % frequency, indicating ongoing daily consumption despite COVID-19 measures. Methamphetamine and morphine's consumption dipped then rose around club reopening, hinting at limited access. The consumption trend of methadone appeared to compensate for the use of morphine. Ketamine and NPS demonstrated similar patterns throughout the entire period. NPS as party drugs seemed influenced by an unstable supply chain and complexities in implementation. Benzodiazepines, commonly abused alongside synthetic cathinones in Taiwan exhibited an opposing trend to NPS while aligned with acetaminophen, suggesting elevated stress and anxiety levels during the pandemic. No significant differences were observed in drug consumption between weekdays and weekends, potentially indicating that COVID-19 measures blurred the traditional distinctions between these timeframes. Environmental ImplicationNew psychoactive substances refer to chemically modified variants of controlled drugs designed to mimic the effects of the original drugs while evading modern detection methods, categorizing them as hazardous materials. The study presents a sewage monitoring project conducted from 2021 to 2024, collecting samples from four WWTPs to analyze NPS and conventional drug trends during and after the COVID-19 pandemic. The findings uncovered connections between drug consumption patterns and pandemic-related policies. In light of the persistent drug abuse and their environmental presence, the results bear critical importance for both environmental and public health. We provide a thorough assessment of these relationships and prioritize areas for future research.

Chemical characteristics and formation mechanism of secondary inorganic aerosols: The decisive role of aerosol acidity and meteorological conditions

In recent years, there has been a growing concern about air pollution and its impact on the air quality and human health, especially for fine particulate matter (PM2.5) and its associated secondary aerosols in urban areas. This study conducted a year-long field campaign to collect PM2.5 samples day and night in an urban area of central Taiwan. Higher PM2.5 mass concentrations were observed in winter (27.7 ± 9.7 μg/m3), followed by autumn (22.5 ± 8.3 μg/m3), spring (19.2 ± 6.4 μg/m3), and summer (11.0 ± 3.1 μg/m3). The dominant formation mechanism of secondary inorganic aerosols was heterogeneous reactions of NO3− at night and homogeneous reactions of SO42− during the day. Additionally, significant correlations were observed between aerosol liquid water content (ALWC) and NO3− during nighttime, indicating the importance of aqueous-phase NO3− formation. The role of aerosol acidity was explored and a unique alkaline condition was found in spring and summer, which showed lower PM2.5 concentrations than the neutralized condition. Under the neutralized condition, higher PM2.5 concentrations were commonly found when combining the ammonium-rich regime with molar ratios of [NO3−]/[SO42−] exceeding 1.6, suggesting the importance of reducing both NH3 and NOx. Furthermore, the results showed that reducing NH3 should be prioritized under high temperature conditions, while reducing NOx became important under low temperature conditions. Clustering of backward trajectories showed that long-range transport could enhance the formation of secondary aerosols, but local emissions emerged as the main factor driving high PM2.5 concentrations. This study provides insights for policymakers to improve air quality, suggesting that different mitigation strategies should be formulated based on meteorological variables and that using clean energy for vehicles and electricity generation is important to alleviate air pollution.

STUDY OF THE EFFECTIVENESS OF THE APPLICATION OF COMPLEX GEOLOGICAL AND TECHNOLOGICAL MEASURES ON THE EXAMPLE OF GAS-CONDENSATE GENERATION IN THE CENTRAL PART OF THE PRIAXIAL ZONE OF THE DNIPROV-DNETZ BASIN

The field development system is the basis of hydrocarbon production, as it ensures the achievement of the necessary technical and economic indicators. Therefore, a high-perfor- mance development system is necessary to ensure maximum profit in the shortest possible time. Empirical studies, analysis and assessment of the effect of implementing a complex of geological and technological measures on the operational facility of a gas condensate field in the central part of the peri-axial zone of the Dnieper-Donets Basin were carried out in order to find methods for improving the existing gas and gas condensate field development systems and increasing gas extraction rates. It is shown that due to hydraulic fracturing, drilling and transfer of wells to the production facility during the five years of production of the produced gas, it is 7.8 % more than in the previous 21 hours, the rate of development of the field from 2015 to 2019 increased from 2.52 %. to 7.9 %, the average well yield increased by 83 % with an increase in the operational fund by 55 %. The results of studies of the effect of the application of geo- logical-technological measures are shown in the form of graphical dependences of the change in the rate of sampling on the number of production wells over time, the rate of daily gas sampling on the number of conducted geological-technological measures on the researched operational object over time. It has been determined that the main driver of the increase in gas production is the hydraulic fracturing of the formation in most of the operational wells, thanks to which the average increase in the output of the wells is 425 %. According to the results of geological and industrial data processing and analysis of changes in the rate of daily gas extraction over time, the effect of the performed operations is divided into three time segments lasting 15–18 months. The decrease in the duration of the effect from the increase in the fund of production wells and the conduct of hydraulic fracturing of the reservoir is due to the decrease in the volume of reservoir energy, therefore the investigated complex of measures will have the greatest effect when implemented at the initial stage of the development of a reservoir with similar geological parameters.

Large-scale performance assessment of the BG-Counter 2 used with two different mosquito traps

BackgroundMosquitoes are important vectors of pathogens. They are usually collected with CO2-baited traps and subsequently identified by morphology. This procedure is very time-consuming. Automatic counting traps could facilitate timely evaluation of the local risk for mosquito-borne pathogen transmission or decision-making on vector control measures, but the counting accuracy of such devices has rarely been validated in the field.MethodsThe Biogents (BG)-Counter 2 automatically counts mosquitoes by discriminating the size of captured objects directly in the field and transmits the data to a cloud server. To assess the accuracy of this counting device, 27 traps were placed at 19 sampling sites across Germany and used in daily, weekly or bimonthly intervals from April until October 2021. The BG-Counter 2 was attached to a CO2-trap (BG-Pro trap = CO2-Pro) and the same trap was converted to also attract gravid mosquitoes (upside-down BG-Pro trap with a water container beneath = CO2-Pro-gravid). All captured mosquitoes were identified by morphology. The number of females (unfed and gravid), mosquito diversity and the number of identified specimens in relation to the counting data of the BG-Counter were compared between the two trapping devices to evaluate sampling success and counting accuracy.ResultsIn total 26,714 mosquitoes were collected during 854 trap days. The CO2-Pro-gravid trap captured significantly more mosquitoes per trap day for all specimens, gravid females and non-gravid females, while there was no difference in the mosquito diversity. The linear model with the captured mosquitoes as a response and the counted specimens as a predictor explained only a small degree of the variation within the data (R2 = 0.16), but per individual trap the value could reach up to 0.62 (mean R2 = 0.23). The counting accuracy for the daily samples had a significant positive correlation with sample size, resulting in higher accuracy for the CO2-Pro-gravid trap and higher accuracy for sites and sampling months with high mosquito abundance.ConclusionsWhile the accuracy of the BG-Counter 2 is quite low, the device is able to depict mosquito phenology and provide information about local population dynamics.Graphical

Enrichment of Phosphates, Lead, and Mixed Soil‐Organic Particles in INPs at the Southern Great Plains Site

AbstractIce nucleating particles (INPs) are rare particles that initiate primary ice formation, a critical step required for subsequent important cloud microphysical processes that ultimately govern cloud phase and cloud radiative properties. Laboratory studies have found that organic‐rich dusts, such as those found in soils, are more efficient INPs compared to mineral dust. However, the atmospheric relevance of these organic‐rich dusts are not well understood, particularly in regions with significant agricultural activity. The Agricultural Ice nuclei at the Southern Great Plains field campaign (AGINSGP) was conducted in rural Oklahoma to investigate how soil dusts contribute to INP populations in the Great Plains. We present chemical characterization of ambient and ice crystal residual particles from a single day of sampling, using single particle mass spectrometry (SPMS) and scanning microscopy. Ambient particles were primarily carbonaceous or secondary aerosol, while the fraction of dust particles was higher in the residual particles. We also observed an unusual particle type consisting of a carbonaceous core mixed with dust fragments on the surface, which was found in higher proportion in residuals. Dust particles measured during residual sampling contained greater proportions of phosphate (63 and 79) and lead (206Pb+). Strong sulfate signals were not seen in the residual dust particles measured by the SPMS, while nitrate was slightly depleted relative to ambient dust. This study shows that organic‐rich soils may be important contributors to the ambient INP population in agricultural regions.

Stability of ascorbic acid in human aqueous humor

ObjectiveThe stability of ascorbic acid (AA) in the human aqueous humor (AqH) remains unclear. This study aimed to investigate the stability of AqH AA under varying conditions (27, 4, − 20, and − 80 °C) without acidification.ResultsRapid AA degradation occurred at 27 °C. At 4 °C, a significant 12.2% degradation was observed after 24 h. Storage at − 20 °C resulted in a notable 37.5% degradation after 28 days, whereas storage at − 80 °C resulted in 10.7% degradation after 28 days. Unacidified AqH samples recorded early decomposition at 27 °C and 4 °C. In conclusion, it is recommended to conduct measurements within 28 days for samples stored at − 80 °C.

Modification of Anaerobic Digestion Model No. 1 for modeling anaerobic digestion of cattle manure with changing solids retention time

In this study, Anaerobic Digestion Model No.1 (ADM1) was modified to incorporate changes in biochemical parameters due to solids retention time (SRT) variations. Cattle manure (CM) and thermally hydrolyzed CM were selected for testing. Continuous anaerobic digestion reactors were operated under different SRT conditions ranging from 6.6 to 36.0 days for both samples. The biochemical parameters (kch, kli, kpr, um,ac, um,bu, um,pro, um,va, Kac, Kbu, Kpro, and Kva) for each SRT condition were determined. To modify ADM1, the equations obtained through linear regression were substituted into biochemical parameters as a function of SRT. The modified ADM1 demonstrated superior accuracy compared with conventional ADM1. This study implies the feasibility of optimizing biochemical parameters for modeling in response to changes in environmental variables.

The impact of gastrointestinal parasitism on the behaviour and welfare of weaned housed lambs

Gastrointestinal (GI) parasitism is a health and production concern in sheep, yet its impact on animal welfare remains unclear. The impact of subclinical infections is especially ambiguous as GI parasitism often remains undiagnosed until clinical signs such as diarrhoea are evident. This study applied quantitative and qualitative methods to examine the effects of subclinical Teladorsagia circumcincta infection on the behaviour and welfare of 96 Suffolk-cross lambs (24 pens of 4 lambs) weaned at 10 weeks old. The hypothesis that parasitism causes negative affective states was tested. Lambs were divided into three groups at the pen level: ad-lib fed control (AC), restricted-fed control (RC), and ad-lib fed parasitised (AP). Parasitised lambs (AP) were dosed three times weekly with 7000 third stage T. circumcincta larvae (L3) from 16 weeks of age. Lambs in the RC group were pair fed to match AP feed intake to separate the effects of infection-induced anorexia from the potential direct impacts of infection. From 7 days pre-infection to 23 days post-infection, scan and behaviour samples were taken from video recordings to quantitatively monitor behaviour, and animal-based measures such as faecal soiling score (FSS) were recorded as welfare indicators. Lying, standing, eating, play and social behaviour were monitored. Qualitative behaviour assessment (QBA) was conducted weekly using the AWIN (2015) protocol to gain insight into the lambs’ affective states over the onset of infection. Parasitised lambs were more likely to stand inactive than AC lambs as the infection progressed (P=0.006). They were also less likely to display eating behaviour in the third daily scan sample than RC lambs (P<0.001). Principal Component Analysis of the QBA data revealed that the first dimension (PC1) described arousal levels, the second (PC2) described the valence of the animals’ affective states, and the third (PC3) described fearfulness and aggression levels. Parasitised lambs (est=10.64,SE=0.33) scored higher than RC lambs (est=9.42, SE=0.33) on PC3, the fearfulness dimension (P=0.030). There were no differences between fearfulness scores of AC and AP lambs or RC lambs and treatment group had no significant impact on the distribution of scores on PC1 or PC2. These findings demonstrate that subclinical GI parasitism negatively impacts lamb welfare not only in the health domain but in the behaviour and mental domains as well. This has implications for welfare assessments and early disease detection in lambs. Future research could explore remote monitoring of the indicators of parasitism identified in this study.

Tracking the prevalence and emergence of SARS-CoV-2 variants of concern using a regional genomic surveillance program.

In our manuscript, we describe the results of SARS-CoV-2 genomic surveillance from February 2021 to June 2022 at a metropolitan hospital in the United States. We demonstrate that consistent daily sampling is sufficient to track the regional prevalence and emergence of variants of concern (VOCs). Similar sampling efforts should be considered a viable option for local SARS-CoV-2 genomic surveillance at other regional laboratories. While the SARS-CoV-2 pandemic has evolved into a more endemic form, we still believe that additional real-world information about sampling, procedures, and data interpretation is valuable for ongoing as well as future genomic surveillance efforts. Our study should be of substantial interest to clinical virologists.

National-scale antimicrobial resistance surveillance in wastewater: A comparative analysis of HT qPCR and metagenomic approaches

Wastewater serves as an important reservoir of antimicrobial resistance (AMR), and its surveillance can provide insights into population-level trends in AMR to inform public health policy. This study compared two common high-throughput screening approaches, namely (i) high-throughput quantitative PCR (HT qPCR), targeting 73 antimicrobial resistance genes, and (ii) metagenomic sequencing. Weekly composite samples of wastewater influent were taken from 47 wastewater treatment plants (WWTPs) across Wales, as part of a national AMR surveillance programme, alongside 4 weeks of daily wastewater effluent samples from a large municipal hospital. Metagenomic analysis provided more comprehensive resistome coverage, detecting 545 genes compared to the targeted 73 genes by HT qPCR. It further provided contextual information critical to risk assessment (i.e. potential bacterial hosts). In contrast, HT qPCR exhibited higher sensitivity, quantifying all targeted genes including those of clinical relevance present at low abundance. When limited to the HT qPCR target genes, both methods were able to reflect the spatiotemporal dynamics of the complete metagenomic resistome, distinguishing that of the hospital and the WWTPs. Both approaches revealed correlations between resistome compositional shifts and environmental variables like ammonium wastewater concentration, though differed in their interpretation of some potential influencing factors. Overall, metagenomics provides more comprehensive resistome profiling, while qPCR permits sensitive quantification of genes significant to clinical resistance. We highlight the importance of selecting appropriate methodologies aligned to surveillance aims to guide the development of effective wastewater-based AMR monitoring programmes.