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The therapeutic potential of plant extracts has attracted significant interest, especially regarding indigenous species with health-promoting properties. Gymnema inodorum , native to Northern Thailand, is recognized for its rich phytochemical profile; however, the impact of various extraction techniques on its phenolic composition and bioactivity remains underexplored. Optimizing extraction methods is essential to enhance the pharmacological efficacy of this plant's bioactive compounds. This study investigated the influence of four extraction methods-ethanol maceration, ethanol reflux, aqueous decoction, and microwave-assisted extraction-on the bioactive profile of G. inodorum leaves, with a focus on the phenolic content and biological activities. Antioxidant activities were evaluated using DPPH, ABTS, and FRAP assays, while the total phenolic and flavonoid contents were quantified by colorimetric methods. High-Performance Liquid Chromatography (HPLC) quantified gymnemic acid and key phenolic compounds. Among the methods, ethanol reflux yielded the highest antioxidant activities (DPPH and ABTS scavenging), with a total phenolic content of 82.54 mg GAE/g and flavonoid content of 31.90 mg QE/g. HPLC analysis identified sinapic acid, myricetin, and p-hydroxybenzoic acid as major phenolics. Furthermore, the ethanol reflux extract displayed potent anti-diabetic activity, with IC 50 values of 13.36 mg/mL for α-amylase and 7.39 mg/mL for α-glucosidase, as well as strong anti-inflammatory activity (IC 50 of 1.6 mg/mL) and acetylcholinesterase inhibition (IC 50 of 1.2 mg/mL). These findings suggest that ethanol reflux extraction is a highly effective method for producing bioactive-rich G. inodorum extracts, with substantial pharmacological potential for developing herbal remedies and nutraceuticals, particularly in enhancing therapeutic approaches for diabetes and other health-related conditions.

Considering the limited information on the impact of PM 2.5 content on ocular health, a follow-up study was conducted on 50 healthy adults. Samples were collected twice, once before the PM 2.5 exposure season and again after exposure. Daily PM 2.5 concentration data was gathered from Thung Satok monitoring station. All subjects completed the self-structured ocular symptom questionnaire. The concentrations of 1-OHP were determined using HPLC-FLD. Logistic regression analysis investigated the relationship between PM 2.5 toxicity and ocular symptoms. The findings revealed that daily PM 2.5 concentrations surpassed the WHO-recommended range by around threefold. Exposure to PM 2.5 significantly raised the likelihood of ocular redness (adjusted OR: 12.39, 95% CI), watering (adjusted OR: 2.56, 95% CI), and dryness (adjusted OR: 5.06, 95% CI). Additionally, these symptoms had an exposure-response relationship with increasing 1-OHP levels. Ocular symptoms worsened in frequency and severity during the high PM 2.5 season, showing a strong link to elevated PM 2.5 levels. Lymphocyte counts were also positively correlated with redness, watering, and dryness during high PM 2.5 exposure. In conclusion, our study shows that subjects exposed to higher PM 2.5 levels presented more significant ocular surface alterations., (© 2024. The Author(s).)

The aim was to study associations between dialkylphosphates (DAPs), organophosphate (OP) metabolites in urine, biomarkers of OP insecticide exposure, and respiratory symptoms among children in upper northern Thailand. We recruited junior high school children in randomly selected schools in four cities (N = 337), with repeated data collection in wet and dry seasons. Urine was collected and analyzed for six OP metabolites, with creatinine adjustment. Total DAP was expressed as sum of DAPs. Data on respiratory symptoms was collected by a standardized questionnaire. Associations were analyzed by multiple logistic regression. Totally 11.3 % lived in farm families. Total DAPs concentration was higher in dry season (p = 0.002) but did not differ between farm and non-farm children. Total DAPs in wet season was associated with current wheeze (p = 0.019), current asthma attacks (p = 0.012) and attacks of breathlessness in last 12 months (p = 0.021). Total DAPs in dry season was associated with current wheeze (p = 0.042), and associations between DAPs and respiratory symptoms were stronger for dimethylphosphate metabolites (DMPs) than for diethylphosphate metabolites (DEPs). DMPs are produced by certain OP pesticides. Biomass burning inside or outside the home, and dampness or mold at home, enhanced the association between total DAPs and attacks of breathlessness. In conclusion, OP pesticide exposure, measured as urinary DAPs, was higher in dry season and similar in farm and non-farm children. OPs exposure, especially to DMP related pesticides, can increase asthmatic symptoms, especially in wet season. Combined exposure to OP and smoke from biomass burning, or dampness and mold, can further increase the prevalence of attacks of breathlessness. There is a need to reduce OP insecticide and biomass smoke exposure among Thai children. Since different pesticides can be used in different seasons, studies on respiratory health effects of OPs pesticide exposure should be done in different seasons., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

The absence of high-affinity antibodies has hindered the development of satisfactory immunoassays for dichlorvos (DDVP) and trichlorfon (TCP), two highly toxic organophosphorus pesticides. Herein, the de novo synthesis of a novel anti-DDVP hapten was introduced. Subsequently, a specific anti-DDVP monoclonal antibody (Mab) was produced with satisfying affinity to DDVP (IC 50 : 12.4 ng mL -1 ). This Mab was highly specific to DDVP, and TCP could readily convert into DDVP under mild alkaline conditions. Leveraging this insight, an indirect competitive ELISA was successfully developed for simultaneous detection of DDVP and TCP. The limit of detection in rice, cabbage and apple for DDVP /TCP was found to be 12.1/14.6 μg kg -1 , 7.3/8.8 μg kg -1 and 6.9/8.3 μg kg -1 , respectively. This study not only provides an effective strategy for producing a high-quality anti-DDVP Mab but also affords a reliable and cost-effective tool suitable for high-throughput detection of DDVP and TCP in food samples., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

The widespread use of organophosphate (OP) and carbamate (CM) pesticides requires efficient and cost-effective detection methods. This study introduces a micro-electrometric method using cricket cholinesterase (ChE) to detect OP and CM residues, providing a rapid and economical alternative to conventional chromatographic techniques. The parameters of the method, including the substrate concentration, incubation temperature, and incubation time, were optimized. By leveraging the sensitivity of cricket ChE to OP and CM inhibition, this approach translates enzyme inhibition into an electrical signal to quantify pesticide levels, achieving an impressive limit of detection (LOD) from 0.036 to 0.086 parts per million (ppm). This method demonstrated reproducibility and stability, making it suitable for field applications and on-site testing across various environmental matrices. This research represents a significant advancement in pesticide residue analysis with potential applications in the development of portable biosensor devices for real-time environmental monitoring and public health protection., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Wongta et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Honey bee brood (HBB) ( Apis mellifera L.), a traditional protein source, has been studied for its nutritional value, but bio-functional properties and safety concerns have not been verified. This study examined the Antioxidant capacity, phytochemicals, minerals, and chemical pollutants in worker broods from several apiaries in Northern Thailand. HBB samples were lyophilized to evaluate antioxidant capacity using ABTS, DPPH, and FRAP assays, tests with water, and 70% ethanol extracts. Phytochemicals were identified using LC-QTOF-MS; pollutants were analyzed chromatographically, and minerals were determined using ICP-OES. The results showed that the evaluated antioxidant capacity of the ethanol extracts included DPPH 2.04-3.37 mg/mL, ABTS 21.22-33.91 mg/mL, and FRAP 50.07-104.15 mg AAE/100 g dry weight. Water extracts had outstanding antioxidant activities except for ABTS, with DPPH 10.67-84.97 mg/mL, ABTS 9.25-13.54 mg/mL, and FRAP 57.66-177.32 mgAAE/100 g dry weight. Total phenolics and flavonoids in ethanol extracts ranged from 488.95-508.87 GAE/100 g to 4.7-12.98 mg QE/g dry weight, respectively. Thirteen phytochemicals were detected and contained adequate mineral contents in the HBBs from different locations found, which were K, Ca, Mg, and Na, and no heavy metals or pollutants exceeded safe levels. These results imply that HBB from different apiaries in Northern Thailand is a nutritious food source with considerable antioxidants and a safe and sustainable food source.

The immunological sexing method using antibodies offers cost-effective, high-volume production but faces challenges in terms of X-sperm purity in sexed semen. This research aimed to produce sexed bull semen using highly specific recombinant antibodies in magnetic-activated cell sorting (MACS), evaluate sperm quality and kinematic parameters, and verify the sex ratio of sperm, embryos, and live calves. Fresh semen from two Angus bulls was separated into two equal groups: conventional (CONV) semen and semen sexed using MACS with Y-scFv antibody conjugation to separate two fractions, i.e., the X-enriched and Y-enriched fractions. Then, computer assisted semen analysis and imaging flow cytometry were used to evaluate sperm motility and kinematic variables, acrosomal integrity, sperm viability, and sperm sex ratios. The results showed that sperm motility and quality did not differ between X-enriched and CONV semen. However, the Y-enriched fraction showed significantly lower sperm quality than the X-enriched fraction and CONV semen. The sperm ratio revealed that X-sperm accounted for up to 79.50% of the X-enriched fraction, while Y-sperm accounted for up to 78.56% of the Y-enriched fraction. The sex ratio of embryos was examined using in vitro fertilization. The cleavage rates using CONV and X-enriched semen were significantly higher than that using Y-enriched semen. Accordingly, 88.26% female blastocysts were obtained by using X-enriched semen, and 83.58% male blastocysts were obtained by using Y-enriched semen. In farm trials, 304 cows were subjected to AI using X-enriched and CONV semen. The pregnancy rate did not differ between the X-enriched and CONV semen groups. On the other hand, X-enriched semen generated significantly more live female calves (83.64%) than CONV semen (47.00%). The MACS sexing method significantly enhanced the X-sperm purity in sexed semen, producing high-quality sperm, a high percentage of female blastocytes, and a high percentage of live female calves., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Understanding heavy metals in rivers is crucial, as their presence and distribution impact water quality, ecosystem health, and human well-being. This study examined the presence and levels of nine heavy metals (Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, and Zn) in 16 surface water samples along the Chao Phraya River, identifying Fe, Mn, Zn, and Cr as predominant metals. Although average concentrations in both rainy and dry seasons generally adhered to WHO guidelines, Mn exceeded these limits yet remained within Thailand's acceptable standards. Seasonal variations were observed in the Chao Phraya River, and Spearman's correlation coefficient analysis established significant associations between season and concentrations of heavy metals. The water quality index (WQI) demonstrated varied water quality statuses at each sampling point along the Chao Phraya River, indicating poor conditions during the rainy season, further deteriorating to very poor conditions in the dry season. The hazard potential index (HPI) was employed to assess heavy metal contamination, revealing that during the dry season in the estuary area, the HPI value exceeded the critical threshold index, indicating the presence of heavy metal pollution in the water and unsuitable for consumption. Using the species sensitivity distribution model, an ecological risk assessment ranked the heavy metals' HC5 values as Pb > Zn > Cr > Cu > Hg > Cd > Ni, identifying nickel as the most detrimental and lead as the least toxic. Despite Cr and Zn showing a moderate risk, and Cu and Ni posing a high risk to aquatic organisms, the main contributors to ecological risk were identified as Cu, Ni, and Zn, suggesting a significant potential ecological risk in the Chao Phraya River's surface water. The results of this study provide fundamental insights that can direct future actions in preventing and managing heavy metal pollution in the river ecosystem., (© 2024. The Author(s).)

Background: At present, porcine reproductive and respiratory syndrome (PRRS) caused by the PRRS virus (PRRSV) is one of the most severe epidemics impacting pig farming globally. Despite the fact that a number of studies have been conducted on potential solutions to this problem, none have proven effective. The focus of problem solving is the use of natural ingredients such as plant extracts. Popular throughout Asia, Caesalpinia sappan (CS) is a therapeutic plant that inhibits PRRSV in vitro. Therefore, this study was performed to determine the efficacy of CS extract dietary supplementation on the productive performance, antibody levels, immunological indicators, and lung pathology of PRRSV-challenged weaned pigs. A total of 32 weaned piglets (28 days old) were randomized into 4 groups and kept separately for 14 days. The treatments were organized in a 2 × 2 factorial design involving two factors: PRRSV challenge and supplementation with 1 mg/kg CS extract. The pigs in the PRRSV-challenged groups were intranasally inoculated with 2 mL of PRRSV (VR2332) containing 10 4 TCID 50 /mL, while those in the groups not challenged with PRRSV were inoculated with 2 mL of normal saline., Results: In the PRRSV-challenged group (CS + PRRSV), supplementation with CS extract led to an increase in white blood cells (WBCs) on Day 7 post infection (p < 0.05) and particularly in lymphocytes on Days 7 and 14. The antibody titer was significantly greater in the CS + PRRSV group than in the PRRSV-challenged group not administered CS (PRRSV group) on Day 14 postinfection (S/P = 1.19 vs. 0.78). In addition, CS extract administration decreased the prevalence of pulmonary lesions, which were more prevalent in the PRRSV-challenged pigs that did not receive the CS extract., Conclusion: The findings of this study suggest that supplementation with CS extract is beneficial for increasing WBC counts, especially lymphocytes, increasing the levels of antibodies and reducing the prevalence of lung lesions in PRRSV-infected pigs., (© 2024. The Author(s).)

In this study, the efficacy of ionomers and poly-lactic acid (PLA) as an alternative solid material combined with scFv antibodies specific to bovine Y-sperm (Y-scFv) was studied to create a novel method of sexing technology. The coupling efficiency of Y-scFv to the surface of PLA, Na + and Zn 2+ ionomer film was between 2 and 8 mg/mL. Fourier transform infrared spectra confirm that Y-scFv was bound with a carboxylic acid group in each film. Therefore, Na + , Zn 2+ ionomers and PLA films conjugated with 4 and 8 mg/mL Y-scFv showed the highest concentration of Y-sperm in the eluted fraction. Considering that the elute fraction was enriched Y-sperm fraction, it contained 67.70-77.94 % of the Y-sperm ratio related to the produced supernatant fraction, which contained up to 69.31-76.01 % enriched X-sperm. In addition, the sperm quality after the sexing process was analyzed by CASA and imaging flow cytometry, which showed that each polymer did not have a negative effect on sperm motility and acrosome integrity for X-sperm. The capacity of ionomer and PLA combined with Y-scFv are used for bovine sperm sexing., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Korawan Sringarm reports financial support was provided by Chiang Mai University. Korawan Sringarm has patent #2101007525 pending to Licensee., (Copyright © 2023 Elsevier B.V. All rights reserved.)

This study aims to determine the prevalence of microorganisms and antibiotic-resistant microorganisms in beehives located on different plantations in Thailand. Seventeen swabs immersed in transport media were utilized for samples from different zones within beehives. Traditional microbial culture-based methods, biochemical tests, MALDI-TOF MS (VITEK ® MS, bioMerieux, Marcy-l'Étoile, France), and antibiotic drug susceptibility (disk-diffusion) tests were used to detect microorganism and antimicrobial resistance bacteria. The results from 16 beehive swabs found Gram-positive bacteria at 59.5%, Gram-negative bacteria at 35.1%, and fungi (yeast) at 5.4%. These organisms are classified as 11, 11, and 2 types of Gram-positive bacteria, Gram-negative bacteria, and fungi (yeast), respectively. Furthermore, no organism showed resistance to vancomycin or cefoxitin for antibiotic drug susceptibility testing. In contrast, all Acinetobacter spp. were susceptible to ciprofloxacin, levofloxacin, ceftazidime, cefotaxime, imipenem, and meropenem, except for Acinetobacter schindleri , which was resistant to ceftazidime and cefotaxime. For other organisms, due to the limitations of tests to identify some environmental microbial species, the antimicrobial susceptibility test results cannot be interpreted as resistant or susceptible to the drug for these organisms. The study's findings will support prevention, healthcare services, and public health systems.

Traditional plants have played a significant role in human culture and medicine throughout history. These plants have the capability to synthesize a diverse range of chemical compounds that serve essential biological functions. This study's objective was to analyze the phytochemical composition of five traditional plants, namely Emilia sonchifolia , Chloranthus erectus , Caesalpinia mimosoides , Acacia concinna , and Tacca chantrieri , native to northern Thailand, using LC-QTOF/MS analysis and assess their potential bioactivity through various assays including DPPH radical scavenging activity, ABTS radical scavenging activity, ferric ion reducing antioxidant power, total phenolic compounds, and total flavonoid content. The findings revealed the presence of natural bioactive compounds in each plant extract, which exhibited pharmacological activity. Notably, Caesalpinia mimosoides displayed the highest antioxidant capacity across all plant extracts (IC 50 in DPPH with the methanol extract was 0.03 and 898.18 mg AAE/100 g with the ethanol extract), along with elevated levels of total phenolic and flavonoid content, which showed the highest TFC at 46.79 µgRE/g in the methanol extract. In conclusion, traditional plants possess notable biological constituents and antioxidant properties, suggesting their potential for bioactive applications. Based on these findings, these indigenous plants can serve as a valuable resource in traditional medicine, offering the possibility of uncovering new products with similar capabilities and additional therapeutic attributes worthy of future exploration.

We demonstrate that dopamine can be used as a reagent for colorimetric enzyme-linked immunosorbent assay (ELISA) using horseradish peroxidase (HRP). Dopamine was able to be polymerized in the presence of HRP and H 2 O 2 , and black polydopamine was obtained after the enzymatic reaction. Because of the black color, the absorbance was significantly changed in the whole range of the visible light region. Here, an indirect competitive ELISA based on the polymerization of dopamine was performed to detect a fluoroquinolone antibiotic, enrofloxacin. The antibiotic is commonly used in livestock farming. The anti-antibiotics antibody was produced from egg yolk from chicken hens. In the visible range, sufficient absorbance changes of ∼0.4∼0.5 and a low background level for the ELISA response were obtained, and the 50 % inhibitory concentration value at 450 nm was determined to be 26 ppb. The performance of the indirect competitive ELISA based on the polymerization of dopamine was compared to that based on the oxidation of catechol because dopamine has a catechol skeleton. By the complex of HRP and H 2 O 2 , catechol can be oxidized to o -benzoquinone having a maximum absorption wavelength of 420 nm. It was shown that the absorbance change in the case of polydopamine was about 2.5 times higher than that of catechol, where the background levels were similar. This confirms that the polymerization of dopamine significantly enhanced the photosignal., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Bee propolis has been touted as a natural antimicrobial agent with the potential to replace antibiotics. Numerous reports and reviews have highlighted the functionalities and applications of the natural compound. Despite much clamor for the downstream application of propolis, there remain many grounds to cover, especially in the upstream production, and factors affecting the quality of the propolis. Moreover, geopropolis and cerumen, akin to propolis, hold promise for diverse human applications, yet their benefits and intricate manufacturing processes remain subjects of intensive research. Specialized cement bees are pivotal in gathering and transporting plant resins from suitable sources to their nests. Contrary to common belief, these resins are directly applied within the hive, smoothed out by cement bees, and blended with beeswax and trace components to create raw propolis. Beekeepers subsequently harvest and perform the extraction of the raw propolis to form the final propolis extract that is sold on the market. As a result of the production process, intrinsic and extrinsic factors, such as botanical origins, bee species, and the extraction process, have a direct impact on the quality of the final propolis extract. Towards the end of this paper, a section is dedicated to highlighting the antimicrobial potency of propolis extract.

Farmers are regarded as a high-risk population for pesticide exposure because of the frequency of their occupational exposure to organophosphate (OP) pesticides. The purposes of this study were 1) to compare urinary OP metabolites, nerve conduction, and neurobehavioral (NB) performance between farmers and a control group, as well as between baseline measurement and after spraying OP pesticides; 2) to investigate the factors associated with increased urinary OP metabolites after spraying OP pesticides; and 3) to investigate the effects of OP metabolites on changes in nerve conduction and NB performance after spraying OP pesticides. This study was conducted with a sample size of 71 farmers and a control group of 26. The findings showed that after spraying OP pesticides total dialkylphosphate (DAP) levels were significantly higher than baseline levels. Multiple linear regression showed that increased urinary OP metabolites among farmers after spraying pesticides were negatively associated with wearing a mask when spraying OP pesticides and positively associated with the number of years of farm work and type of sprayer equipment. The results also showed a positive association between the increased urinary total diethylphosphate (DEP) levels and the decreased amplitude of motor nerve conduction. Furthermore, a positive correlation was observed between increased urinary total DEP and decreased right amplitude of sympathetic skin response (SSR). An increase in urinary total DMP was positively associated with a decrease in raw score and a standard score of visual-motor integration (VMI). Our findings provide evidence that exposure to OP pesticides can cause a deficit in nerve conduction and NB performance in farmers. These findings may contribute to the early detection of neurological disease and inform strategies to prevent damage to the peripheral and central nervous systems., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Gymnema inodorum (Lour.) Decne is a vegetable local to Chiang Mai Province of Northern Thailand. This study aimed to analyze the antioxidant and phytochemical potential of G. inodorum found in Chiang Mai Province; antioxidant compounds of G. inodorum were tested via DPPH, ABTS and FRAP assays, and total phenolic compound and total flavonoid contents were analyzed. Anti-inflammatory effects were focused on regarding pharmacological potential. The gymnemic acid level was analyzed by HPLC-UV, and other potential chemicals were analyzed by LC-QTOF/MS. The quantifications of gymnemic acid contents analyzed using HPLC-UV showed that the highest gymnemic acid concentrations were found in the air-dried and roasted 1-day-fermented leaf extracts (0.1258 ± 0.0157 μg/mg). The highest free radical scavenging activity via DPPH assay was found in baked leaf extract, with an IC 50 of 8.99 mg/mL, and via ABTS assay in baked and roasted leaf extracts, with an IC 50 of 1.05 mg/mL. FRAP assays showed the highest free radical scavenging activity for the baked leaf extract, with 0.0085 ± 0.008 mM Fe 2+ /g sample. The total phenolic contents of fresh G. inodorum leaf extracts obtained with ethanol, methanol and water were 0.19 ± 0.0004, 0.21 ± 0.0010 and 0.10 ± 0.0008 μg GAE/g, respectively. The total flavonoid contents of fresh G. inodorum leaf extracts obtained with ethanol, methanol and water were 74.56 ± 28.00, 71.88 ± 16.11 and 10.74 ± 3.63 μg QE/g, respectively. The LC-QTOF/MS analysis of the fresh G. inodorum leaf extract showed that 6-hydroxykaempferol 7-rutinoside was the most abundant compound. In the study, G. inodorum , a plant local to Northern Thailand, is shown to be a useful plant with high antioxidant and phytochemical potential properties.

Ph 3 P-I 2 -mediated condensation reactions of isatoic anhydrides and o -phenylenediamines have been developed for the regioselective syntheses of a wide range of linearly and angularly fused benzoimidazoquinazolinones. The selectivity of the products relies on the generation of either highly electrophilic oxyphosphonium or less reactive imidate intermediates. A direct amine attack at the C-2 position of the oxyphosphonium intermediate presumably drives the reaction toward the linearly fused products, whereas an attack of the diamine at the C-4 position of the in situ generated cyclic imidate leads to the angularly fused derivatives. This strategy serves as a practical handle for the efficient synthesis of other related heterocycles.

Inappropriate use of personal protective equipment (PPE) among pesticide applicators may increase urinary organophosphate (OP) metabolite levels and subsequently increase risks of cognitive performance. Therefore, this study aims to (1) compare urinary OP metabolite levels and cognitive performance between pre-and post-pesticide application seasons; (2) PPE use and factors associated with PPE use linked to increased urinary OP metabolite levels during pesticide application; and (3) the association between urinary OP metabolite levels and cognitive performance. This longitudinal follow-up study on 79 pesticide applicators was carried out between October 2021 and January 2022. The applicators were interviewed, collected urine samples, and tested for cognitive performance in pre-and post-pesticide application seasons. The results found that the levels of urinary OP metabolites in post-application season were significantly higher than those in pre-application season ( p < 0.001). Multiple linear regression analysis found that increased total diethylphosphate (DEP) and total dialkylphosphate (DAP) levels were associated with not wearing gloves while mixing pesticides [beta (β) ± standard error (SE) = -43.74 ± 18.52, 95% confidence interval (95% CI) = -80.84, -6.64 for total DEP and -50.84 ± 19.26, 95% CI = -89.41, -12.26 for total DAP] and also with not wearing a mask while spraying pesticides (β ± SE = -31.76 ± 12.24, 95% CI = -56.28, -7.24 for total DEP and -33.20 ± 12.63, 95% CI = -58.49, -7.92 for total DAP) after adjusting for covariates. The scores of Montreal Cognitive Assessment-Thai, Thai Mental State Examination, and Mini-Cognitive test in post-pesticide application were significantly lower than those in pre-pesticide application ( p < 0.001). However, no association was found between urinary OP metabolite levels and cognitive decline. Our findings indicate that inappropriate PPE use during pesticide application was the major factor affecting urinary OP metabolite levels among pesticide applicators. Wearing gloves when mixing pesticides and a mask when spraying pesticides were key factors in reducing occupational exposure to OP. Exposure to OP at low levels and for short periods of exposure may not affect cognitive performance significantly. Therefore, long-term exposure and exposure to high levels of OP should be investigated further., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Chittrakul, Sapbamrer and Hongsibsong.)

The improper and excessive use in agriculture of chlorpyrifos-methyl (CPSM) and chlorpyrifos-ethyl (CPSE) may affect the health of human beings. Herein, a fluorescence-based immunochromatographic assay (FICA) was developed for the simultaneous determination of CPSM and CPSE. A monoclonal antibody (mAb) with equal recognition of CPSM and CPSE was generated by the careful designing of haptens and screening of hybridoma cells. Instead of labeling fluorescence with mAb, the probe was labeled with goat-anti-mouse IgG (GAM-IgG) and pre-incubated with mAb in the sample. The complex could compete with CPS by coating antigen in the test line. The new format of FICA used goat-anti-rabbit IgG (GAR-IgG) conjugated with rabbit IgG labeled with fluorescence microspheres as an independent quality control line (C line). The novel strategy significantly reduced nonspecific reactions and increased assay sensitivity. Under the optimal conditions, the proposed FICA showed a linear range of 0.015-64 mg/L and limit of detection (LOD) of 0.015 mg/L for both CPSE and CPSM. The average recoveries of CPS from spiked food samples by FICA were 82.0-110.0%. The accuracy was similar to the gas chromatography-tandem mass spectrometry (GC-MS/MS) results. The developed FICA was an ideal on-site tool for rapid screening of CPS residues in foods.

Background: Thailand's agricultural sector is crucial to the country's development and economy. The COVID-19 pandemic caused negative effects on the agricultural context and the mental health of Thai farmers. This study aimed to compare changes in the agricultural context and mental health among farmers in different regions of Thailand during the fifth wave of the COVID-19 pandemic. We also investigated the determinants associated with the mental health of farmers., Methods: This cross-sectional study was carried out from December 2021 to January 2022 in Thailand, with 1,676 Thai farmers interviewed., Results: The results found that the Central region found to be the highest impact of increased agrochemical costs (91.0%) and decreased crop yields (92.0%), while the highest in the North and the North-East was found in decreased crop product prices (84.0 and 66.4%). In the context of economic status, the greatest impact in the Central region was found to be in increased household expense (96.3%), while the highest impact in the North, the North-East, and the South was found in decreased household income (91.5, 80.0, and 69.8%, respectively). Regarding mental health, the Central region was found to be the highest prevalence of extremely high stress and severe depression symptoms (18.3 and 23.4%), whereas the South region was found to be the highest prevalence of severe anxiety symptoms (7.4%). Interestingly, the multivariate analysis also found that the agricultural factors associated with mental health were decreased working days, changes in agrochemical type and crop cultivation, decreased crop rotation, increased difficulty in accessing agrochemicals and markets, decreased crop yields, and increased household debt., Conclusion: The findings of this study are useful for government and relevant organizations to plan and implement supportive measures and improve mental health services to mitigate both short and long-term impacts of the COVID-19 pandemic suit on the regions. Specific measures and facilities should be tailored toward the target regions., (© 2022. The Author(s).)

Fluoroquinolones (FQs) are among the antibiotics whose widespread use in farm-raised animals results in potentially harmful residues in the end products. Additionally, most Thai farmers use antibiotics. Amoxicillin and enrofloxacin were commonly used by pig farms, and hens were given enrofloxacin to prevent immunization side effects. Moreover, antibiotic overuse has harmed food safety in the long term, and the use of low-dose antibiotics causes bacterial resistance. Herein, an indirect competitive enzyme-linked immunosorbent assay (icELISA) was used to make a fast, easy, sensitive, and cost-effective method for monitoring FQs residues. After immunizing hens with mixed multi-hapten ciprofloxacin-bovine serum albumin (CPFX-BSA) with norfloxacin-bovine serum albumin (NFX-BSA), the IgY antibody purified from egg yolk was used for the detection of FQs residues in chicken and pork samples. The efficiency of the IgY antibody showed excellent sensitivity, with 50% inhibitory concentration (IC 50 ) of enrofloxacin at 0.05 µg/mL, far below the MRLs defined by the EU for muscle samples, which was not to exceed 100 µg/kg. The recovery range for chicken muscle samples spiked with ENFX concentrations of 1.00-0.01 µg/mL was 86.65-112.71%, similar to pork samples, which were 84.24-117.22.2%. This method has a lot of potential for analyzing fluoroquinolones in complex samples quickly, easily, and at a low cost on-site. The IgY-based ic ELISA was developed to detect ciprofloxacin (CPFX), norfloxacin (NFX), and enrofloxacin (ENFX) residues; it confirms that IgY could be a promising choice for the detection of antibiotic residues in food samples.

The Smilacaceae family has been used as a food source and herbal medicine for a long time. This study aims to identify the phytochemicals extracted from Smilax glabra and Smilax corbularia by using LC-QTOF/MS analysis and determine their bioactive potential. Compounds were identified from S. glabra and S. corbularia extracts by LC-QTOF-MS and it was found that longistylin A and CAY10435 have higher degrees of matching compounds (99.66% and 99.87%). Smilax glabra showed antioxidant capacity, i.e., DPPH and ABTS at percentage inhibitions of 71.94 ± 1.46% and 59.84 ± 4.80%, respectively, and FRAP at 730.69 ± 33.62 mg AAE/100 g sample. The total phenolic compound contents of the ethanol, methanol, and water extracts were 0.017 ± 0.001, 0.015 ± 0.001, and 0.016 ± 0.001 mg GAE/g, respectively, while the total flavonoid contents were 0.043 ± 0.002, 0.033 ± 0.002, and 0.006 ± 0.003 mg QE/g, respectively. The anti-inflammatory capacity showed 97.26% protection and 2.74% hemolysis. The antimicrobial activity can inhibit Gram-positive bacteria with a minimum inhibitory concentration (MIC) of 62.5 mg/mL and a minimum bactericidal concentration (MBC) of 500 mg/mL. Smilax corbularia showed antioxidant capacity, i.e., DPPH and ABTS at percentage inhibitions of 72.24 ± 0.64% and 39.87 ± 2.37%, respectively, and FRAP at 208.33 ± 50.80 mg AAE/100 g sample. The total phenolic compound contents of the ethanol, methanol, and water extracts were 0.006 ± 0.000, 0.007 ± 0.002, and 0.002 ± 0.001 mg GAE/g, respectively, while the total flavonoid contents of the ethanol and methanol extracts were 0.012 ± 0.001 and 0.008 ± 0.000 mg QE/g, respectively. The anti-inflammatory capacity showed 96.64% protection and 3.36% hemolysis. The antimicrobial activity of the extracts can inhibit Gram-positive bacteria with a MIC of 31.25 mg/mL and MBC of 125 mg/mL for the ethanol extract and a MIC of 125 mg/mL and MBC of 62.5 mg/mL for the methanol extract. In conclusion, Smilax glabra and Smilax corbularia were found to contain several phytochemicals that can be used for further study. Both Smilax species can also be used as sources of antioxidants and herbal medicines for killing Gram-positive bacteria.

In Thailand, pesticides containing organophosphates (OP) are frequently applied to crops to suppress insects. School children can be exposed to OPs on a daily basis, from food consumption to breathing and touching pesticides drifted near classrooms. Living in an agricultural area can also be one of the causes. As a result, it is important to monitor OPs residues in the food chain and biomarkers of exposure. The Gas Chromatography-Flame Photometric Detector method was employed to examine the relationship between OPs residue and DAPs (Diakly phosphate) in four targeted locations in Thailand, as well as to examine the residues of OPs in vegetable samples and DAPs in 395 school children's urine samples. Vegetables were found to contain at least one OP, with chlorpyrifos being the most prevalent. The OPs detected frequencies for Sakon Nakhon, Chiang Mai, Phang Nga, and Pathum Thani are 96.1%, 94%, 91.7%, and 83.3%, respectively. The overall centration level of OPs showed 0.3261 mg/kg, 0.0636 mg/kg, 0.0023 mg/kg, 0.0150 mg/kg, 0.2003 mg/kg, 0.0295 mg/kg, and 0.0034 mg/kg for diazinon, dimethoate, pirimiphos-methyl, chlorpyrifos, profenofos, ethion, and triazophos, respectively. Nearly 98% of school children were detected with at least one DAP. The overall level of dimethyl phosphate metabolites (5.258 µmole/mole creatinine) in urine samples is higher than diethyl phosphate metabolites (2.884 µmole/mole creatinine), especially in the case of Pathum Thani. Our findings show a consistent relationship between OPs in vegetables from wet markets and DAPs in urine samples of school children in various parts of Thailand.

Early detection of decline in neurobehavioral (NB) performance requires reliable methods of testing. Although NB tests have been shown to be consistent and reliable in Western countries, there has been limited research in Asian populations., Objective: The purpose of this study was to investigate the test-retest reliability of NB tests in a Thai adult population and examine the impact of demographic data on NB tests. The aspects of the tests chosen were memory, attention, hand-eye coordination, motor speed, and dexterity., Methods: The three NB tests used were digit span, Purdue Pegboard, and visual-motor integration. All three were administered to a population of 30 Thai adults., Results: The outcomes of all Pearson's correlation coefficient tests (r) were positive and greater than 0.60, and subtest-retest reliability correlation coefficients ranged from 0.63 (p<0.001) to 0.81 (p<0.001). Interestingly, the outcomes of all of these tests were not affected by demographic data, with the exception of the Purdue Pegboard test, in which performance on the preferred hand and both hands assessment was weakly associated with age (β=-0.09, p<0.001 and β=-0.08, p<0.05, respectively)., Conclusions: NB tests have adequate reliability and are useful for the evaluation of clinical memory, attention, hand-eye coordination, motor speed, and dexterity in Thai adults. These tests were not affected by demographic data. However, further studies to measure the validity of the digit span, Purdue Pegboard, and visual-motor integration tests are needed., Competing Interests: Disclosure: The authors report no conflicts of interest.

Distinguishing between bull Y- and X-bearing sperm populations is advantageous for techniques using sexed bull semen. The aim of this study was to produce a single-chain fragment variable (scFv) antibody against plasma membrane epitopes on bull Y-bearing sperm. Variable heavy (VH)- and variable light (VL)-region genes generated from a hybridoma cell secreting a specific Y-bearing sperm monoclonal antibody (mAb-1F9) were cloned and expressed. The expected sizes of the DNA bands were ∼350 bp for the VH gene and ∼318 bp for the VL gene. The VH and VL genes were generated and used to construct an scFv gene (∼650 bp), which was expressed in E.coli TG1 cells and produced the corresponding soluble scFv antibody. Compared with the parent mAb-1F9, the scFv antibodies presented a high affinity for Y-bearing sperm and low cross-reactivity with X-bearing sperm. An immunofluorescence analysis confirmed that the scFv antibodies and mAb-1F9 recognize epitopes on the Y-bearing sperm surface. The fluorescence signal was strong on the plasma membrane of Y-bearing sperm but very weak for X-bearing sperm. This study aids the application and production of engineered scFv antibodies specific to Y-bearing sperm to distinguish between Y- and X-bearing sperm populations for techniques involving sexed bull semen.

Sperm sexing technique is favored in the dairy industry. This research focuses on the efficiency of bovine sperm sexing using magnetic-activated cell sorting (MACS) by scFv antibody against Y-chromosome-bearing sperm (Y-scFv) coupled to magnetic microbeads and its effects on kinematic variables, sperm quality, and X/Y-sperm ratio. In this study, the optimal concentration of Y-scFv antibody coupling to the surface of magnetic microbeads was 2-4 mg/mL. PY-microbeads revealed significantly enriched Y-chromosome-bearing sperm (Y-sperm) in the eluted fraction (78.01-81.43%) and X-chromosome-bearing sperm (X-sperm) in the supernatant fraction (79.04-82.65%). The quality of frozen-thawed sexed sperm was analyzed by CASA and imaging flow cytometer, which showed that PY-microbeads did not have a negative effect on X-sperm motility, viability, or acrosome integrity. However, sexed Y-sperm had significantly decreased motility and viability. The X/Y-sperm ratio was determined using an imaging flow cytometer and real-time PCR. PY-microbeads produced sperm with up to 82.65% X-sperm in the X-enriched fraction and up to 81.43% Y-sperm in the Y-enriched fraction. Bovine sperm sexing by PY-microbeads showed high efficiency in separating Y-sperm from X-sperm and acceptable sperm quality. This initial technique is feasible for bovine sperm sexing, which increases the number of heifers in dairy herds while lowering production expenses.

An ideal theoretical model for personal protective clothing (PPC) is to protect wearers from external workplace hazards while meeting certain ergonomic requirements. This study aims to compare the efficiency of different personal protective clothing in its protection against chlorpyrifos, including gum rosin-coated PPC, commercial PPC (Tychem ® coverall), and everyday clothing, during application under field conditions. Perception of discomfort and physiological effects after wearing PPC were also investigated. Thirty-one applicators were invited and consented to participate in the study. The study established that the median percentage of chlorpyrifos protection efficiency was 90.7% for commercial PPC, 89.2% for gum rosin-coated PPC, and 76.5% for everyday PPC. When the protection efficiency was compared among the different types of PPC, the percentage protection efficiency of gum rosin-coated PPC was not significantly different from that of commercial PPC. The percentage protection efficiencies of commercial PPC and gum rosin-coated PPC were significantly higher than that of everyday PPC. The major characteristics of gum rosin-coated PPC were water-repellency, breathability, low cost, and ease of manufacture. Therefore, this study suggests that gum rosin-coated PPC is a reasonable alternative PPC for farmers, particularly in low and middle-low-income countries and tropical climates, to protect against pesticides while providing acceptable comfort.

Objective: This cross-sectional study aimed to examine farmers' knowledge, awareness, practices regarding pesticide use, and prevalence of health symptoms related to pesticides exposure among farmers who applied organophosphates (OP) and pyrethroids (PY)., Methods: Data regarding demographic variables and health symptoms pertinent to pesticide use was collected from 67 farmers who applied OP and 50 farmers who applied PY using interviews from January to March 2021., Results: The farmers who applied OP had lower knowledge, awareness, and prevention practices regarding pesticide use than those who applied PY. After adjustment of covariate variables, the farmers who applied OP had a significantly higher prevalence of respiratory conditions (OR = 8.29 for chest pain, OR = 6.98 for chest tightness, OR = 27.54 for dry throat, and OR = 5.91 for cough), neurological symptoms (OR = 10.62 for fatigue and OR = 6.76 for paresthesia), and neurobehavioral symptoms (OR = 13.84 for poor concentration, OR = 3.75 for short term memory, and OR = 8.99 for insomnia) related to pesticide exposure than those who applied PY., Conclusion: Our findings suggest that OP had a more adverse effect on human health than PY, resulting in a higher prevalence of pesticide-related symptoms. The outcomes of this study have the benefit of providing vital information for all stakeholders with regard to the implementation of safe practices in the utilization of personal protective equipment (PPE) and pesticide use in a health intervention and health promotion program., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Thammachai, Sapbamrer, Rohitrattana, Tongprasert, Hongsibsong and Wangsan.)

Background: Measurements of urinary dialkyl phosphate (DAP) metabolites are often used to characterize exposures to organophosphate (OP) insecticides; however, some challenges to using urinary DAP metabolites as an exposure measure exist. OP insecticides have short biological half-lives with measurement in a single urine sample typically only reflecting recent exposure within the last few days. Because of the field staff and participant burden of longitudinal sample collection and the high cost of multiple measurements, typically only one or two urine samples have been used to evaluate OP insecticide exposure during pregnancy, which is unlikely to capture an accurate picture of prenatal exposure., Methods: We recruited pregnant farmworker women in Chom Thong and Fang, two districts of Chiang Mai province in northern Thailand (N = 330) into the Study of Asian Women and their Offspring's Development and Environmental Exposures (SAWASDEE) from 2017 to 2019. We collected up to 6 serial urine samples per participant during gestation and composited the samples to represent early, mid, and late pregnancy. We measured concentrations of urinary DAP metabolites in the composited urine samples and evaluated the within- and between-participant variability of these levels. We also investigated predictors of OP insecticide exposure., Results: DAP metabolite concentrations in serial composite samples were weakly to moderately correlated. Spearman correlations indicated that composite urine samples were more highly correlated in Fang participants than in Chom Thong participants. The within-person variances (0.064-0.65) exceeded the between-person variances for DETP, DEP, ∑DEAP, DMP, DMTP, ∑DMAP, ∑DAP. The intraclass correlations (ICCs) for the volume-based individual metabolite levels (ng/mL) ranged from 0.10 to 0.66. For ∑DEAP, ∑DMAP, and ∑DAP the ICCs were, 0.47, 0.17, 0.45 respectively. We observed significant differences between participants from Fang compared to those from Chom Thong both in demographic and exposure characteristics. Spearman correlations of composite samples from Fang participants ranged from 0.55 to 0.66 for the ∑DEAP metabolite concentrations in Fang indicating moderate correlation between pregnancy periods. The ICCs were higher for samples from Fang participants, which drove the overall ICCs., Conclusions: Collecting multiple (∼6) urine samples during pregnancy rather than just 1 or 2 improved our ability to accurately assess exposure during the prenatal period. By compositing the samples, we were able to still obtain trimester-specific information on exposure while keeping the analytic costs and laboratory burden low. This analysis also helped to inform how to best conduct future analyses within the SAWASDEE study. We observed two different exposure profiles in participants in which the concentrations and variability in data were highly linked to the residential location of the participants., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

This quasiexperimental study was designed to determine the effectiveness of a participatory program on pesticide use behavior and blood cholinesterase levels. The participants were 18-60 years old, living in Thung Satok subdistrict, San Pa Tong District, Chiang Mai Province. Eighty subjects, including 32 farmers and 48 consumers, were recruited to participate in the study voluntarily by a purposive sampling technique. Data concerning each individual's behaviors were collected using questionnaires, and blood cholinesterase levels were obtained from whole blood finger, providing whole blood pre and postexperiment. The data were analyzed using Fishers' exact and paired t -tests, for the problem situations were independently analyzed. The results show that the participatory process made it possible to improve methods for the prevention of the unhealthy effects of pesticides. After participating in the activities, both groups showed significantly higher scores in before and after knowledge ( P < 0.05) and a decrease in pesticide contamination in their bodies as a result of the measurement of average cholinesterase which increased significantly ( P < 0.001). In conclusion, the participatory program was applied to solve health problems from chemical hazards. The program can raise awareness about chemical toxicity, leading to the modification of the related behavior toward chemicals and reduction of the contamination of chemicals in the body. Therefore, the adoption of participatory processes can effectively solve problems related to chemical hazards that affect health., Competing Interests: The authors declare that there are no conflicts of interest., (Copyright © 2021 Nootchakarn Sawarng et al.)

Organophosphate (OP) pesticides may accumulate in pregnant agricultural workers, resulting in adverse effects on the growth and development of the fetus and neonates. This study aims to evaluate a possible association between prenatal urinary OP metabolite levels among pregnant agricultural workers and birth outcomes of infants. This study also investigated the factors associated with urinary OP metabolites among pregnant agricultural workers. The spot urine samples were collected and analyzed for six OP metabolite levels. Birth outcomes data were abstracted from medical records. Multiple regression analysis found that gestational age at childbirth was negatively associated with diethylphosphate (DEP) levels (β = -0.073; 95% CI, -0.121, -0.024). Apgar score at 1 and 5 min after birth were negatively associated with diethyldithiophosphate (DEDTP) levels (β = -0.036; 95% CI, -0.069, -0.003; and β = -0.034, 95% CI, -0.057, -0.011, respectively). In addition, DEDTP levels were negatively associated with maternal age (β = -0.181; 95% CI, -0.339, -0.023), and dimethylphosphate (DMP) levels were positively associated with frequency of agricultural work during pregnancy (β = 31.554; 95% CI, 0.194, 62.914). Our results indicate that prenatal OP exposure can cause adverse birth outcomes in babies. Therefore, it is necessary to develop an effective strategy for reducing prenatal exposure to OP pesticides., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Coffee pulp is one of the most underutilised by-products from coffee processing. For coffee growers, disposing of this agro-industrial biomass has become one of the most difficult challenges. This study utilised this potential biomass as raw material for polyphenolic antifungal agents. First, the proportion of biomass was obtained from the Arabica green bean processing. The yield of by-products was recorded, and the high-potency biomass was serially extracted with organic solvents for the polyphenol fraction. Quantification of the polyphenols was performed by High Performance Liquid Chromatography (HPLC), then further confirmed by mass spectrometry modes of the liquid chromatography-quadrupole time-of-flight (QTOF). Then, the fraction was used to test antifungal activities against Alternaria brassicicola , Pestalotiopsis sp. and Paramyrothecium breviseta . The results illustrated that caffeic acid and epigallocatechin gallate represented in the polyphenol fraction actively inhibited these fungi with an inhibitory concentration (IC50) of 0.09, 0.31 and 0.14, respectively. This study is also the first report on the alternative use of natural biocontrol agent of P. breviseta , the pathogen causing leaf spot in the Arabica coffee.

Carbamate pesticides (CPs) are the most used pesticides in agricultural production and pest control. In this study, carbofuran, isoprocarb and carbaryl were employed as models, and a general hapten strategy based on carbamate moiety recognition was proposed. Molecular modeling of the three-dimensional (3D) structure and surface electrostatic potential of the CPs indicated that the amide group formed by conjugation significantly influenced recognition by antibodies. The proposed strategy was used to obtain three sensitive and specific monoclonal antibodies (mAbs) with IC 50 values of 1.4 ng/mL, 8.4 ng/mL and 13.8 ng/mL for carbofuran, isoprocarb and carbaryl, respectively. Negligible cross-reactivity (%) with analogs was observed, except for fenobucarb (84.6%) for isoprocarb. The obtained antibodies were used to develop an immunochromatographic assay (ICA) to simultaneously and quantitatively detect the three CPs. A strip reader was used to determine the limits of quantitation (LOQs) as 0.05 ng/mL (carbofuran), 31.3 ng/mL (isoprocarb) and 31.3 ng/mL (carbaryl). The recoveries of cucumber and Chinese cabbage samples ranged from 76% to 111%, with CVs from 1.3% to 10.6%, indicating good potential for the rapid simultaneous detection of multiple pesticide residues in a large batch of samples., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Porcine reproductive and respiratory syndrome virus (PRRSV) is a major epidemic in pig production, leading to economic losses in the pig industry worldwide. The use of medicinal plants with antiviral properties might be useful help to prevent and control PRRSV outbreaks. Caesalpinia sappan (CS) heartwood is an important herbal ingredient used in Thai folk medicine, possessing various biological activities, including antiviral activity. The present study focuses on the in vitro antiviral activity against PRRSV of a semi-purified fraction of ethanolic CS crude extract using preparative high-performance liquid chromatography. Qualification of the fractions illustrating positive antiviral activity was carried out with liquid chromatography-quadrupole time-of-flight mass spectrometry. The preparative chromatography separated the crude extract into six consecutive fractions, among which the first fraction showed potential antiviral activity by inhibiting PRRSV replication in a MARC-145 monolayer (virus titer 2.75 median tissue culture infective dose (TCID 50 )/mL (log 10 ) vs. 9.50 median log10 TCID 50 /mL of the control) at 72 h post-infection, and this fraction included byakangelicin, brazilin, naringenin, and brazilein. These results provide useful information for further study to effectively develop the CS bioactive antiviral compounds against PRRSV as a feed additive or veterinary drug in the pig industry.

Organophosphate (OP) pesticides can transfer from mother to fetus via the placenta and amniotic fluid and may affect the development of infants. This study aims to evaluate the associations between maternal OP concentrations collected in the 1st-2nd trimester and the 3rd trimester of pregnancy and the infant developmental performance. The Screening Test of the Bayley Scales of Infants and Toddler Development, Third Edition (BSID-III screening test) was used to assess development performance at 2 and 6 months of age. Multiple regression analysis showed a negative correlation between cognitive performance at 2 months and maternal diethylthiophosphate (DETP) levels in the 1st-2nd trimester ( β ± SE = -0.012 ± 0.004, p < 0.05). We also found that expressive communication and fine motor performance at 6 months were negatively associated with maternal diethyldithiophosphate (DEDTP) levels in the 3rd trimester ( β ± SE = -0.047 ± 0.016, p < 0.05, and β ± SE = -0.044 ± 0.017, p < 0.05, respectively). These results suggest that maternal ethylated OP concentrations at different timing of exposure during pregnancy may influence different aspects of infant developmental performance.

Farmers are at a high risk of inhalation exposure when handling pesticides. Thai farmers usually protect themselves against pesticide exposure by wearing commercial respiratory protective equipment (RPE) available from rural community markets. However, scientific data regarding the pesticide filtration efficiency of RPE commonly worn by farmers is limited. Thus, this study aimed to investigate the efficiency of insecticide filtration of various RPE commonly worn by farmers in Thailand. The half facepiece respirator was used as a control to compare the results with other RPE. Ten types of RPE were selected for testing. The filtration efficiency of each RPE against insecticides was tested in a laboratory. The remarkable findings were that a surgical mask demonstrated the least filtration efficiency of all tested insecticides, with a range of 25.7-61.5%. The RPE available in rural markets of Thailand had a filtration efficiency within a range of 64.9-95.4%, whereas a half facepiece respirator was the most efficient in filtering insecticides, with a range of 96.5-98.9%. Therefore, our results suggest that the RPE most frequently worn by farmers may not provide adequate protection when compared with the respirator. However, considerations around RPE use in low-and middle-income countries and tropical climate conditions should be based on pesticide toxicity and practical use, ensuring balance between the risks from pesticide exposure and acceptance of PPE use.

Amyloid beta peptides (A β 1-42) have been found to be associated with the cause of Alzheimer's disease (AD) and dementia. Currently, methods for detecting A β 1-42 are complicated and expensive. The present study is aimed at developing an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) to detect A β 1-42 by using a polyclonal antibody from alpaca, an application used in urine samples. The serum was collected from the alpaca after immunizing it with A β 1-42 at 500  μ g/injection 5 times. The ic-ELISA was developed and showed a half-maximal inhibitory concentration (IC 50 ) of 103.20 ng/ml. The limit of detection (LOD) was 0.39 ng/100  μ l. The cross-reactivity was tested with A β 1-40 and 8 synthesized peptides that had sequence similarities to parts of A β 1-42. The cross-reactivity of A β 1-40 and peptide 1 (DAEFRHDSGYE) was 55% and 69.4%, respectively. The ic-ELISA was applied to analyze A β 1-42 in the urine and precipitated protein urine samples. This method can be used for detecting a normal level of total soluble A β (approximately 1 ng in 5 mg of precipitated urine protein) and can be used for detecting the early stages of AD. It is considered to be an easy and inexpensive method for monitoring and diagnosing AD., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2020 Anurak Wongta et al.)

A novel phosphonium-mediated reaction of isatins is described. In the presence of alcohol, the reaction proceeds to furnish C-12 modified tryptanthrin derivatives. Without alcohol, self-dimerization of isatins gives rise to tryptanthrin and its analogs. This divergent and step-economic approach provides a facile access to diverse indoloquinazoline structures including the natural alkaloids, methylisatoid and cephalanthrin B, in high yields from simple precursors under mild and metal-free reaction conditions.

Insulin resistance is a risk factor for various diseases. Chronic organophosphate exposure has been reported to be a cause of insulin resistance in animal models. This cross-sectional study aimed to evaluate the association between organophosphate exposure and insulin resistance in pesticide sprayers and nonfarmworkers. Participants aged 40-60 years, consisting of 150 pesticide sprayers and 150 nonfarmworkers, were interviewed and assessed for their homeostatic model assessment of insulin resistance (HOMA-IR) level. Organophosphate (OP) exposure was measured in 37 sprayers and 46 nonfarmworkers by first morning urinary dialkyl phosphate (DAP) metabolites. The DAP metabolite levels were not different in either group except for diethylthiophosphate (DETP; p = 0.03), which was higher in sprayers. No significant association was observed between DAP metabolite levels and HOMA-IR. Wearing a mask while handling pesticides was associated with lower dimethyl metabolites (95% CI = -11.10, -0.17). Work practices of reading pesticide labels (95% CI = -81.47, -14.99) and washing hands after mixing pesticide (95% CI = -39.97, -3.35) correlated with lower diethyl alkylphosphate level. Overall, we did not observe any association between OP exposure and insulin resistance in pesticide sprayers and the general population. However, personal protective equipment (PPE) utilization and work practice were associated with OP exposure level in sprayers.

A flow enzyme-linked immunosorbent assay (ELISA) method based on light absorption by enzymatically generated aniline oligomer in the presence of horseradish peroxidase (HRP), H 2 O 2 , and aniline is proposed. Aniline oligomer is rapidly formed through the polymerization reaction via the enzymatic reaction, and its fast reaction rate is beneficial for flow ELISA. An anti-3-phenoxybenzoic acid monoclonal antibody (mAb) was produced by mice, and was used for the flow competitive ELISA for the determination of 3-phenoxybenzoic acid (3PBA), which was performed on an acrylic plate having a Y-shaped channel. ABS resin beads (d = 1 mm) were filled in the channel to increase the surface area for the adsorption of the mAb. A clank-type detection chamber (optical length: 1 cm) made of polydimethylsiloxane (PDMS) containing carbon black, which can significantly decrease light scattering, was fabricated with a 3D printer. The PDMS detection chamber was connected to the outlet of the acrylic flow chip with a tube. A blue LED was used as a light source for the flow ELISA. The inhabitation concentration at 50% and the detection range (absorbance change from 90 to 10%) for the proposed flow competitive ELISA were 0.5 ppm and 0.05-5 ppm, respectively. We also performed the flow competitive ELISA in an artificial and real urine, and no significant matrix effect of the urine samples on the ELISA was found., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Raw materials used for black garlic (BG) processing were collected from the major garlic production areas in Northern Thailand. Five of those were identified as of Thai origin (accession G1-G5), and accession G6 was of the Chinese variety. They were initially analyzed for varietal differences using morphological characteristics and genetic variation. Fresh materials from each accession were dried to the same moisture content (55%-60%) and BG processed at 75°C, 90% relative humidity (RH) for 15 days. Thereafter, physiochemical and chemical profiles were analyzed and compared. The dendrogram from random amplified polymorphic DNA fingerprints grouped G2, G3, G4, and G5 as closely related while G1 and G6 were out-groups. Prior to BG processing, the pH of fresh garlic was approximately 6.3 and decreased to 3.7, thereafter. The contents of chemical properties were independent with genotypes. BG processing improved phenolic, flavonoid, and antioxidant but the content of thiosulfinate was minimized in all BG samples. Overall, result indicated that garlics grown in Northern Thailand were genotypically variable. BG processing altered physical and chemical appearance, and these changes were independent with the genotypes., Competing Interests: The authors declare no conflict of interest with respect to the research., (© 2020 The Authors. Food Science & Nutrition published by Wiley Periodicals LLC.)

Exposure to organophosphate (OP) pesticides in children may increase oxidative stress, resulting in the development of chronic diseases. This study aims to compare urinary OP metabolites and oxidative stress between children in agricultural and urban communities. The study also investigated the factors associated with urinary OP metabolites among children. Urine samples were collected from children for measuring levels of OP metabolites, glutathione (GSH), and malondialdehyde (MDA). The remarkable findings were that total dialkylphosphate (DAP) levels detected among children in this agricultural community were significantly higher than those from the urban community (P = 0.001). Multiple linear regression showed that total diethylphosphate (DEP) levels among children in the agricultural community were negatively associated with distances from children's residence to the agricultural fields (β ± SE. = - 1.535 ± 0.334, 95%CI = - 2.202, - 0.863) and positively associated with playing on farms (β ± SE. = 0.720 ± 0.342, 95%CI = 0.036, 1.405). In addition, total dimethylphosphate (DMP) levels were positively associated with children working on farms (β ± SE. = 0.619 ± 0.264, 95%CI = 0.091, 1.147). Importantly, GSH levels among children in the agricultural community were significantly lower than those in the urban community (P < 0.001), but MDA levels did not differ. These results therefore suggest that children can be exposed to OPs both outdoors and indoors. Our results also provide supporting evidence that OP exposure can cause oxidative stress in children. As oxidative stress contributes to several chronic diseases, a good proposed strategy for the future would include measurement of oxidative stress biomarkers among children exposed to OPs as an early warning of chronic diseases.

Chlorpyrifos is an organophosphate pesticide that is wildly used among farmers for crop protection. However, there are concerns regarding its contamination in the environment and food chain. In the present study, an in-house indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) specific for detecting chlorpyrifos is developed and validated against gas chromatography-flame photometric detection (GC-FPD) as the conventional method. The developed ic-ELISA was used for detecting chlorpyrifos residue in vegetable samples. The developed ic-ELISA showed good sensitivity to chlorpyrifos at an IC 50 of 0.80 µg/kg, with low cross-reactivity to other organophosphate pesticides. The 160 samples were collected from local markets located in the Chiang Rai, Chiang Mai, and Nan provinces in northern Thailand. The positive rate of chlorpyrifos residues in the vegetable samples was 33.8%, with the highest levels found in cucumbers, coriander, and morning glory, at 275, 145, and 35.3 µg/kg, respectively. The highest median levels of chlorpyrifos found in the detected samples were Chinese cabbage (332 μg/kg), cucumber (146.3 μg/kg) and Chinese Kale (26.95 μg/kg). The developed ic-ELISA is suitable for the rapid quantitation of chlorpyrifos residues.

Wearing appropriate personal protective equipment during the application of pesticides is one method of reducing dermal exposure to pesticides. Thus, the aim of this research is to develop personal protective clothing (PPC) coated with gum rosin and investigate the efficiency of its level of protection against chlorpyrifos and cypermethrin. Comparison of the protection efficiency of each PPC with Tychem ® C coveralls was also investigated. Five commercially available cotton fabrics were chosen for tailoring the PPC, and then, the PPC was coated with a gum rosin finish to provide water repellence. The efficiency of the level of protection of the gum rosin-coated PPC against insecticides was tested in a laboratory (closed chamber). The remarkable findings were that the % protection efficiencies for all the PPC, with the exception of one, were not significantly different to those for Tychem ® C coveralls. The protection efficiencies ranged from 99.85% to 99.97% against chlorpyrifos and 99.11% to 99.89% against cypermethrin. Therefore, our results suggest that gum rosin-coated clothing provided satisfactory levels of protection against insecticides and could be considered as suitable protective clothing for pesticide applicators. Choice of an appropriate fabric for coating with gum rosin also needs to be considered. A further study in field conditions is warranted to confirm the protection efficiency in a working environment.