Immune response enhancement by dietary supplementation with Caesalpinia sappan extract in weaned pigs challenged with porcine reproductive and respiratory syndrome virus.

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 104 TCID50/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.

Divergent Approach for Regioselective Synthesis of Linearly and Angularly Fused Benzoimidazoquinazolinones from Isatoic Anhydrides.

Ph3P-I2-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.

Changes in agricultural context and mental health of farmers in different regions of Thailand during the fifth wave of the COVID-19 pandemic.

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.

Production of single-chain fragment variable (scFv) antibodies specific to plasma membrane epitopes on bull Y-bearing sperm.

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.

Divergent Synthesis of Methylisatoid and Tryptanthrin Derivatives by Ph3P-I2-Mediated Reaction of Isatins with and without Alcohols.

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.

In vitro screening antiviral activity of Thai medicinal plants against porcine reproductive and respiratory syndrome virus.

BACKGROUND: Porcine reproductive and respiratory syndrome (PRRS) caused by PRRS virus (PRRSV) results in economic losses in the swine industry globally. Several studies have investigated the use of plant extracts in the prevention and control of PRRS outbreaks. Thai medicinal plants may be useful for treating PRRSV infection in pigs. Therefore, we investigated the in vitro anti-PRRSV and antioxidant properties of seven Thai medicinal plants: Caesalpinia sappan Linn., Garcinia mangostana Linn., Houttuynia cordata, Perilla frutescens, Clinacanthus nutans, Phyllanthus emblica, and Tiliacora triandra. RESULTS: Using antiviral screening, we observed that T. triandra extract strongly inhibited PRRSV infectivity in MARC-145 cells [virus titer 3.5 median tissue culture infective dose (TCID50)/ml (log10)] at 24 h post-infection, whereas C. sappan extract strongly inhibited PRRSV replication [virus titer 2.5 TCID50/ml (log10)] at 72 h post-infection. C. sappan extract had the highest total phenolic content [220.52 mM gallic acid equivalent/g] and lowest half-maximal inhibitory concentration [1.17 mg/ml in 2,2-diphenyl-1-picrylhydrazyl and 2.58 mg/ml in 2,2-azino-bis (3-ethylbenzothiazo-line-6-sulfonic acid) diammonium salt]. CONCLUSION: T. triandra extract could inhibit PRRSV infectivity, whereas C. sappan extract was the most effective in inhibiting PRRSV replication in MARC-145 cells. This study elucidates the antiviral activities of Thai medicinal plant extracts in vivo. The results promise that Thai medicinal plant extracts, particularly T. triandra and C. sappan extracts, can be developed into pharmaceutical drugs for the prevention of PRRS in pigs.

A permethrin metabolite is associated with adaptive immune responses in Gulf War Illness.

Gulf War Illness (GWI), affecting 30% of veterans from the 1991 Gulf War (GW), is a multi-symptom illness with features similar to those of patients with autoimmune diseases. The objective of the current work is to determine if exposure to GW-related pesticides, such as permethrin (PER), activates peripheral and central nervous system (CNS) adaptive immune responses. In the current study, we focused on a PER metabolite, 3-phenoxybenzoic acid (3-PBA), as this is a common metabolite previously shown to form adducts with endogenous proteins. We observed the presence of 3-PBA and 3-PBA modified lysine of protein peptides in the brain, blood and liver of pyridostigmine bromide (PB) and  PER (PB+PER) exposed mice at acute and chronic post-exposure timepoints. We tested whether 3-PBA-haptenated albumin (3-PBA-albumin) can activate immune cells since it is known that chemically haptenated proteins can stimulate immune responses. We detected autoantibodies against 3-PBA-albumin in plasma from PB + PER exposed mice and veterans with GWI at chronic post-exposure timepoints. We also observed that in vitro treatment of blood with 3-PBA-albumin resulted in the activation of B- and T-helper lymphocytes and that these immune cells were also increased in blood of PB + PER exposed mice and veterans with GWI. These immune changes corresponded with elevated levels of infiltrating monocytes in the brain and blood of PB + PER exposed mice which coincided with alterations in the markers of blood-brain barrier disruption, brain macrophages and neuroinflammation. These studies suggest that pesticide exposure associated with GWI may have resulted in the activation of the peripheral and CNS adaptive immune responses, possibly contributing to an autoimmune-type phenotype in veterans with GWI.

Organophosphorus pesticide residues in vegetables from farms, markets, and a supermarket around Kwan Phayao Lake of Northern Thailand.

This study investigated organophosphorus (OP) residues in vegetables from 27 farms, 106 markets, and 1 supermarket around Kwan Phayao Lake, Northern Thailand, between August and September 2013. Types of vegetables sampled were all vegetables cultivated or sold around the study site. The most common OP pesticides detected in farm samples were chlorpyrifos (50 %), malathion (31.8 %), monocrotophos (31.8 %), diazinon (13.6 %), omethoate (13.6 %), and dicrotophos (9.1 %). The most common OP pesticides detected in market samples were chlorpyrifos (33.9 %), diazinon (18.6 %), parathion-methyl (3.4 %), profenofos (3.4 %), primiphos-ethyl (3.4 %), and fenitrothion (1.7 %). The OP pesticides detected in supermarket samples were chlorpyrifos (33.3 %), and diazinon (66.7 %). Among the compounds detected, chlorpyrifos was detected in most of the vegetable samples from all sources. The highest chlorpyrifos level in farm samples were found in lemon balm (2.423 mg/kg) followed by Vietnamese coriander (0.835 mg/kg), and cowpea (0.027 mg/kg). The highest level in markets samples were found in garlic (7.785 mg/kg) followed by Chinese cabbage (2.864 mg/kg) and Vietnamese coriander (1.308 mg/kg). Residues from supermarket samples were found only in parsley (0.027 mg/kg). The findings showed that 16 samples (59.3 %) from farms and 14 samples (13.2 %) from markets contained OP residues at or above the maximum residue limits established by the European Union. It is concluded that awareness, safety education, and strict regulation of pesticide use are necessary.

Advancing micro-electrometric techniques for the detection of organophosphate and carbamate residues using cricket cholinesterase.

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.

Organophosphate insecticide exposure and respiratory symptoms among school children in Northern Thailand: Interaction by biomass burning, dampness and season.

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.

New insights from poly-lactic acid and ionomer films coupled with recombinant antibodies for processing sexed-sorting bovine sperm.

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 Zn2+ 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+, Zn2+ 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.

Sperm quality variables of sex-sorted bull semen produced by magnetic-activated cell sorting coupled with recombinant antibodies targeting Y-chromosome-bearing sperm.

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.

Antioxidant Capacity, Phytochemicals, Minerals, and Chemical Pollutants in Worker Honey Bee (Apis mellifera L.) Broods from Northern Thailand: A Safe and Sustainable Food Source.

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.

De novo synthesis of a novel hapten and development of a monoclonal antibody-based immunoassay for the detection of dichlorvos and trichlorfon.

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 (IC50: 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.

Occurrence, distribution, and ecological risk assessment of heavy metals in Chao Phraya River, Thailand.

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.

Phytochemical Profiling and Antioxidant Capacity of Traditional Plants, Northern Thailand.

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 (IC50 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.

Effects of urinary of organophosphate metabolites on nerve conduction and neurobehavioral performance among farmers in northern Thailand.

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.

Dopamine as a polymerizable reagent for enzyme-linked immunosorbent assay using horseradish peroxidase.

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 H2O2, 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 H2O2, 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.

Exploring the Functional Properties of Propolis, Geopropolis, and Cerumen, with a Special Emphasis on Their Antimicrobial Effects.

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.