A Gram-negative-selective antibiotic that spares the gut microbiome.

Infections caused by Gram-negative pathogens are increasingly prevalent and are typically treated with broad-spectrum antibiotics, resulting in disruption of the gut microbiome and susceptibility to secondary infections1-3. There is a critical need for antibiotics that are selective both for Gram-negative bacteria over Gram-positive bacteria, as well as for pathogenic bacteria over commensal bacteria. Here we report the design and discovery of lolamicin, a Gram-negative-specific antibiotic targeting the lipoprotein transport system. Lolamicin has activity against a panel of more than 130 multidrug-resistant clinical isolates, shows efficacy in multiple mouse models of acute pneumonia and septicaemia infection, and spares the gut microbiome in mice, preventing secondary infection with Clostridioides difficile. The selective killing of pathogenic Gram-negative bacteria by lolamicin is a consequence of low sequence homology for the target in pathogenic bacteria versus commensals; this doubly selective strategy can be a blueprint for the development of other microbiome-sparing antibiotics.

CAMPYLOBACTER HYOINTESTINALIS ISOLATION FROM HOWLER (ALOUATTA CARAYA) AND SPIDER MONKEYS (ATELES FUSCICEPS ROBUSTUS) AT A ZOOLOGIC FACILITY IN CENTRAL ILLINOIS.

Campylobacter hyointestinalis was initially isolated from an asymptomatic black howler monkey (Alouatta caraya) in a routine fecal culture examination. Fecal cultures from other individuals in this group and an adjacently housed black-headed spider monkey (Ateles fusciceps robustus) group recovered C. hyointestinalis from all but one of the individuals sampled (1.1 spider monkeys and 2.1 howler monkeys). Concurrently, one spider monkey presented with acute onset severe rectal prolapse and diarrhea. Whole-genome sequencing results of C. hyointestinalis isolates from all individuals were homologous and closely related to Campylobacter hyointestinalis subsp. hyointestinalis TTU_618, a strain typically associated with environmental samples. In addition, two cytolethal distending toxin (CDT) expressing gene clusters, cdt-I and cdt-II, were identified in all isolates. These results suggest C. hyointestinalis is transmissible to both howler monkeys and spider monkeys, though the origin of infection and whether it is transmissible between these species is undetermined.

Salmonella invasion is controlled through the secondary structure of the hilD transcript.

Virulence functions of bacterial pathogens are often energetically costly and thus are subjected to intricate regulatory mechanisms. In Salmonella, invasion of the intestinal epithelium, an essential early step in virulence, requires the production of a multi-protein type III secretion apparatus. The pathogen mitigates the overall cost of invasion by inducing it in only a fraction of its population. This constitutes a successful virulence strategy as invasion by a small number is sufficient to promote the proliferation of the non-invading majority. Such a system suggests the existence of a sensitive triggering mechanism that permits only a minority of Salmonella to reach a threshold of invasion-gene induction. We show here that the secondary structure of the invasion regulator hilD message provides such a trigger. The 5' end of the hilD mRNA is predicted to contain two mutually exclusive stem-loop structures, the first of which (SL1) overlaps the ribosome-binding site and the ORF start codon. Changes that reduce its stability enhance invasion gene expression, while those that increase stability reduce invasion. Conversely, disrupting the second stem-loop (SL2) represses invasion genes. Although SL2 is the energetically more favorable, repression through SL1 is enhanced by binding of the global regulator CsrA. This system thus alters the levels of hilD mRNA and is so sensitive that changing a single base pair within SL1, predicted to augment its stability, eliminates expression of invasion genes and significantly reduces Salmonella virulence in mice. This system thus provides a possible means to rapidly and finely tune an essential virulence function.

Salmonella Pathogenicity Island 1 Is Expressed in the Chicken Intestine and Promotes Bacterial Proliferation.

Salmonella enterica serovar Enteritidis is a common cause of foodborne illness in the United States. The bacterium can be transmitted to humans via contaminated chicken meat and eggs, and virulence in humans requires type III secretion system 1 (TTSS-1), encoded on Salmonella pathogenicity island 1 (SPI-1). Chickens often carry S Enteritidis subclinically, obscuring the role of SPI-1 in facilitating bacterial colonization. To evaluate the role of SPI-1 in the infection of chicks by Salmonella, we created and utilized strains harboring a stable fluorescent reporter fusion designed to quantify SPI-1 expression within the intestinal tracts of animals. Using mutants unable to express TTSS-1, we demonstrated the important role of the secretion system in facilitating bacterial colonization. We further showed that coinoculation of an SPI-1 mutant with the wild-type strain increased the number of mutant organisms in intestinal tissue and contents, suggesting that the wild type rescues the mutant. Our results support the hypothesis that SPI-1 facilitates S Enteritidis colonization of the chicken and make SPI-1 an attractive target in preventing Salmonella carriage and colonization in chickens to reduce contamination of poultry meat and eggs by this foodborne pathogen.

The protein acyltransferase Pat post-transcriptionally controls HilD to repress Salmonella invasion.

N-Lysine acylation is a post-translational modification important for both prokaryotic and eukaryotic cells to control a wide array of cellular functions. Here we demonstrate that the protein acyltransferase Pat regulates genes on Salmonella Pathogenicity Island 1 (SPI1) that are required for the invasion of the intestinal epithelium. Mutation of pat slightly increased spleen colonization by Salmonella in streptomycin-treated mice, with more of the pat mutant reaching the spleen than the wild type strain. Growth of Salmonella under specific conditions selectively induced expression of Pat, and deletion of pat increased SPI1 gene expression under the same growth conditions. In addition, over-expression of Pat repressed SPI1 expression and bacterial entry into epithelial cells. These results demonstrate that Salmonella invasion is negatively controlled by Pat. Regulation of the SPI1 central regulator HilD was essential for Pat to exert its effects. The control of HilD by Pat was through post-transcriptional mechanisms, moderately repressing hilD translation while significantly reducing HilD stability. Additionally, growth of Salmonella in the presence of histone deacetylases inhibitors reduced expression of SPI1 by affecting HilD stability, supporting the concept that altering the stability of this regulator is required for Pat to control Salmonella invasion.

Bile Acids Function Synergistically To Repress Invasion Gene Expression in Salmonella by Destabilizing the Invasion Regulator HilD.

Salmonella spp. are carried by and can acutely infect agricultural animals and humans. After ingestion, salmonellae traverse the upper digestive tract and initiate tissue invasion of the distal ileum, a virulence process carried out by the type III secretion system encoded within Salmonella pathogenicity island 1 (SPI-1). Salmonellae coordinate SPI-1 expression with anatomical location via environmental cues, one of which is bile, a complex digestive fluid that causes potent repression of SPI-1 genes. The individual components of bile responsible for SPI-1 repression have not been previously characterized, nor have the bacterial signaling processes that modulate their effects been determined. Here, we characterize the mechanism by which bile represses SPI-1 expression. Individual bile acids exhibit repressive activity on SPI-1-regulated genes that requires neither passive diffusion nor OmpF-mediated entry. By using genetic methods, the effects of bile and bile acids were shown to require the invasion gene transcriptional activator hilD and to function independently of known upstream signaling pathways. Protein analysis techniques showed that SPI-1 repression by bile acids is mediated by posttranslational destabilization of HilD. Finally, we found that bile acids function synergistically to achieve the overall repressive activity of bile. These studies demonstrate a common mechanism by which diverse environmental cues (e.g., certain short-chain fatty acids and bile acids) inhibit SPI-1 expression. These data provide information relevant to Salmonella pathogenesis during acute infection in the intestine and during chronic infection of the gallbladder and inform the basis for development of therapeutics to inhibit invasion as a means of repressing Salmonella pathogenicity.

The intestinal fatty acid propionate inhibits Salmonella invasion through the post-translational control of HilD.

To cause disease, Salmonella must invade the intestinal epithelium employing genes encoded within Salmonella Pathogenicity Island 1 (SPI1). We show here that propionate, a fatty acid abundant in the intestine of animals, repressed SPI1 at physiologically relevant concentration and pH, reducing expression of SPI1 transcriptional regulators and consequently decreasing expression and secretion of effector proteins, leading to reduced bacterial penetration of cultured epithelial cells. Essential to repression was hilD, which occupies the apex of the regulatory cascade within SPI1, as loss of only this gene among those of the regulon prevented repression of SPI1 transcription by propionate. Regulation through hilD, however, was achieved through the control of neither transcription nor translation. Instead, growth of Salmonella in propionate significantly reduced the stability of HilD. Extending protein half-life using a Lon protease mutant demonstrated that protein stability itself did not dictate the effects of propionate and suggested modification of HilD with subsequent degradation as the means of action. Furthermore, repression was significantly lessened in a mutant unable to produce propionyl-CoA, while further metabolism of propionyl-CoA appeared not to be required. These results suggest a mechanism of control of Salmonella virulence in which HilD is post-translationally modified using the high-energy intermediate propionyl-CoA.

Dietary monoglyceride supplementation to support intestinal integrity and host defenses in health-challenged weanling pigs.

Frequent incidence of postweaning enterotoxigenic Escherichia coli (ETEC) diarrhea in the swine industry contributes to high mortality rates and associated economic losses. In this study, a combination of butyric, caprylic, and capric fatty acid monoglycerides was investigated to promote intestinal integrity and host defenses in weanling pigs infected with ETEC. A total of 160 pigs were allotted to treatment groups based on weight and sex. Throughout the 17-d study, three treatment groups were maintained: sham-inoculated pigs fed a control diet (uninfected control [UC], n = 40), ETEC-inoculated pigs fed the same control diet (infected control [IC], n = 60), and ETEC-inoculated pigs fed the control diet supplemented with monoglycerides included at 0.3% of the diet (infected supplemented [MG], n = 60). After a 7-d acclimation period, pigs were orally inoculated on each of three consecutive days with either 3 mL of a sham-control (saline) or live ETEC culture (3 × 109 colony-forming units/mL). The first day of inoculations was designated as 0 d postinoculation (DPI), and all study outcomes reference this time point. Fecal, tissue, and blood samples were collected from 48 individual pigs (UC, n = 12; IC, n = 18; MG, n = 18) on 5 and 10 DPI for analysis of dry matter (DM), bacterial enumeration, inflammatory markers, and intestinal permeability. ETEC-inoculated pigs in both the IC and MG groups exhibited clear signs of infection including lower (P < 0.05) gain:feed and fecal DM, indicative of excess water in the feces, and elevated (P < 0.05) rectal temperatures, total bacteria, total E. coli, and total F18 ETEC during the peak-infection period (5 DPI). Reduced (P < 0.05) expression of the occludin, tumor necrosis factor α, and vascular endothelial growth factor A genes was observed in both ETEC-inoculated groups at the 5 DPI time point. There were no meaningful differences between treatments for any of the outcomes measured at 10 DPI. Overall, all significant changes were the result of the ETEC infection, not monoglyceride supplementation.

Infection caused by the bacterium known as enterotoxigenic Escherichia coli (ETEC) is a common disruptor of weaned pigs' health, leading to economic losses for the producers. To determine if nutritional supplementation could help protect against these losses, weaned pigs were assigned to one of three treatments: 1) uninfected and fed a standard nursery pig diet, 2) infected with ETEC and fed the same standard diet, or 3) infected with ETEC and fed the standard diet supplemented with a combination of butyric, caprylic, and capric fatty acid monoglycerides. Growth performance was tracked throughout the 17-d study and health outcomes were measured at the peak and resolution of ETEC infection. At the peak-infection time point, pigs that were infected with ETEC had lower fecal moisture content, greater fecal bacterial concentrations, and elevated body temperatures compared with uninfected pigs. Additionally, infection reduced expression of genes related to inflammation, angiogenesis, and the intestinal barrier during the peak-infection period. Overall, all significant changes were the result of the ETEC infection, and there were no meaningful differences observed between the different treatments.

Human exposures to Brucella canis from a pregnant dog during an international flight: Public health risks, diagnostic challenges and future considerations.

AIMS: This report documents the exposure of passengers and crew of a commercial international flight to the zoonotic pathogen Brucella canis after an infected dog aborted in the passenger cabin of the aircraft. This case demonstrates the challenges associated with brucellosis screening and the risks that airline personnel, airport employees and travellers face when animals with unrecognized zoonotic infections are transported. METHODS/RESULTS: The public health investigation of this case was conducted by the Centers for Disease Control, the Illinois Department of Health and the Illinois Department of Agriculture, in collaboration with a local veterinary clinic and several academic and federal diagnostic laboratories. It included an extensive diagnostic evaluation of the dam and aborted foetuses to confirm a diagnosis of canine brucellosis. Passengers, airline personnel and staff from the veterinary clinic where the dogs were treated underwent risk assessments, and clinic staff also received detailed guidance regarding infection prevention practices. CONCLUSIONS: Animal shelters and breeding programs are recommended to screen dogs routinely for brucellosis, but it is not unusual for domestic or imported animals to have unknown health histories, including the dog's brucellosis status, at the time of purchase, adoption, or re-homing. Testing recommendations and requirements vary by state, making it challenging for state public health and animal health agencies to monitor and respond appropriately. This case highlights the importance of Brucella spp. screening in sexually intact dogs prior to breeding, purchase, or domestic or international transportation of the dogs. The transportation of pregnant dogs may present a previously unrecognized public health threat in addition to contributing to unnecessary stress and health risks for pregnant animals.

Availability of Using Honeybees as Bioindicators of Pesticide Exposure in the Vicinity of Agricultural Environments in Taiwan.

While pollinating, honeybees are subject to exposure to a variety of pesticides; with their characteristics of certain foraging distances, they could serve as bioindicators of pesticide exposure in a neighborhood. We conducted a study to assess availability by collecting and analyzing bee samples from 15 apiaries located in East Taiwan and dust samples from the adjacent environment, and by finding relations between both samples. Seventeen pesticides were selected for the analysis using gas or liquid chromatography coupled with mass spectrometry, and eight (three insecticides, two herbicides, and three fungicides) were more frequently detected from bee or dust samples; the levels of these pesticides were mostly under 1000 ng/g. Significant correlation results (r ≅ 0.8) between residue concentrations in bees and in dust suggest that honeybees could be a good bioindicator for exposure to herbicides and fungicides within certain ranges. The pesticide contents of sick/dead bees were much higher than those of healthy counterparts regarding any pesticide type, with the mean total concentrations of 635 ng/g and 176 ng/g, respectively. We conclude that honeybees could be used as bioindicators of pesticide exposure; sick/dead bees could serve as a warning sign of the severity of pesticide pollution.

Assessing the urinary concentration of nitrofurantoin and its antibacterial activity against Escherichia coli, Staphylococcus pseudintermedius, and Enterococcus faecium isolated from dogs with urinary tract infections.

Nitrofurantoin, a broad-spectrum nitrofuran class antibiotic, is applied as a first-line antibiotic in treating human urinary tract infections (UTIs) due to its great efficacy and high achievable concentration. The interest in using this antibiotic in companion animals has increased due to the growing demand for effective antibiotics to treat UTIs caused by multidrug-resistant bacteria. Currently, the susceptibility interpretations for nitrofurantoin are based on the breakpoints set for humans, while the canine-specific breakpoints are still unavailable. In this study, we assessed the concentration of nitrofurantoin reaching the dog's urine using the recommended oral dosing regimen. In addition, we examined the efficacy of this breakpoint concentration against the common canine UTI pathogens, Escherichia coli, Staphylococcus pseudintermedius, and Enterococcus faecium. Eight experimental beagle dogs were treated with ~5 mg/kg of nitrofurantoin macrocrystal PO 8qh for 7 days. The urine samples were collected via cystocentesis at 2, 4, and 6 h after administration on day 2 and day 7 and used to quantify nitrofurantoin concentrations by ultra-high performance liquid chromatography. The results showed that 26.13-315.87 µg/mL nitrofurantoin was detected in the dogs' urine with a mean and median concentration of 104.82 and 92.75 µg/mL, respectively. Additionally, individual dogs presented with urinary nitrofurantoin concentrations greater than 64 µg/mL for at least 50% of the dosing intervals. This concentration efficiently killed E. coli, and S. pseudintermedius, but not E. faecium strains carrying an MIC90 value equal to 16, 16, and 128 µg/mL, respectively. Taken together, these results suggest that the value of 64 µg/mL may be set as a breakpoint against UTI pathogens, and nitrofurantoin could be an effective therapeutic drug against E. coli and S. pseudintermedius for canine UTIs.

Antimicrobial Resistance in Bacteria Isolated From Canine Urine Samples Submitted to a Veterinary Diagnostic Laboratory, Illinois, United States.

The emergence of antimicrobial resistance (AMR) in dogs constitutes a threat to animal and human health. There is a lack of studies in Illinois that evaluated the prevalence of AMR among urinary bacterial pathogens. In the study, we included 803 isolates (299 Gram-positive and 504 Gram-negative) that were isolated from 2,583 canine urine samples submitted to the Veterinary Diagnostic Laboratory, the University of Illinois between 2019 and 2020 from dogs suspected of urinary tract infections (UTI). The most common Gram-positive isolates included Staphylococcus pseudintermedius (17.93%), Enterococcus faecalis (9.46%), Streptococcus canis (6.10%), and Enterococcus faecium (3.74%), while Gram-negative isolates included Escherichia coli (45.58%), Proteus mirabilis (11.08%), Klebsiella pneumoniae (3.11%), and Pseudomonas aeruginosa (2.99%). Among the Gram-positive isolates, Staphylococcus pseudintermedius isolates showed a very high prevalence of resistance to penicillin (56.94%), a high prevalence of resistance to trimethoprim-sulfamethoxazole (31.94%), enrofloxacin (29.17%), and oxacillin (27.08%). Among Gram-negative bacteria, Escherichia coli isolates showed a high prevalence of resistance to ampicillin (31.42%). Considering the high prevalence of resistance to antimicrobials commonly used to treat UTI in dogs, urine samples should be collected for bacterial culture and susceptibility testing before treatment initiation to prevent treatment failures and the development of multidrug resistance. Given the possibility of zoonotic transmission of antimicrobial-resistant bacteria, veterinarians when treating UTI cases, should inform dog owners of the potential transmission risk.

Development and validation of an insulated isothermal PCR assay for the rapid detection of Mannheimia haemolytica.

We developed a rapid insulated isothermal PCR (iiPCR) assay for on-site detection of Mannheimia haemolytica using a primer and probe set targeting the superoxide dismutase (sodA) gene. Our iiPCR assay detected M. haemolytica clinical isolates successfully and produced negative results on other bovine or ovine respiratory pathogens, including Histophilus somni, Bibersteinia trehalosi, Trueperella pyogenes, Streptococcus suis, and Mycoplasma spp., indicating that the PCR reactions were specific. Additionally, our iiPCR assay detected as few as 21 copies of genomic DNA and 17.2 cfu/mL of bacterial culture, which was 10 and 100 times more sensitive than conventional PCR, respectively. Our iiPCR assay can be performed on a portable device in a total of 58 min and may be a useful tool for the detection of M. haemolytica in bovine and ovine respiratory disease in the field.

Mycobacterium fortuitum abortion in a sow.

Two aborted Chester White pig fetuses were presented to a veterinary diagnostic laboratory in Illinois. Postmortem examination identified no gross abnormalities. Histologic evaluation revealed multifocal necrosis of chorionic epithelial cells, coalescing areas of mineralization in the placenta, and focal accumulations of viable and degenerate neutrophils in the lung. Intra- and extracellular acid-fast bacilli were identified in the lesions in both the placenta and lungs. Bacterial culture of stomach contents yielded heavy growth of Mycobacterium fortuitum, a rapidly growing nontuberculous mycobacterium (NTM), which was further confirmed through whole-genome sequencing. NTM are opportunistic pathogens commonly found in the soil and in contaminated water supplies. In animals, M. fortuitum is typically introduced through cutaneous wounds leading to infections limited to the skin, with systemic infection being uncommon. To our knowledge, abortion caused by M. fortuitum has not been reported previously.

Contribution of Visible Surface Mold to Airborne Fungal Concentration as Assessed by Digital Image Quantification.

The rapid monitoring of total fungi, including air and surface fungal profiling, is an important issue. Here, we applied air and surface sampling, combined with digital image quantification of surface mold spots, to evaluate the contribution of surface fungi to airborne fungal concentrations. Cladosporium, Penicillium, Aspergillus, and yeast often appeared in the air or on wall surfaces during sampling. The indoor/outdoor concentration ratios (I/O ratios) demonstrated that the airborne concentrations of commonly found fungal genera outdoors were higher than those indoors (median I/O ratio = 0.65-0.91), excluding those of Penicillium and yeast. Additionally, the surface density (fungal concentration/area) of individual fungi showed no significant correlation with the airborne concentration, excluding that of Geotrichum. However, if a higher surface ratio (>0.00031) of mold spots appeared in the total area of an indoor environment, then the concentrations of Aspergillus and Geotrichum in the air increased significantly. Our results demonstrated that the airborne concentration of indoor fungi is significantly correlated with the outdoor concentration. A higher density of surface fungi does not necessarily contribute to a high fungal concentration in the air. In contrast to fungal density, quantification of the surface fungal area is recommended to assess the risk of surface fungi propelling into the air.

Associations between phthalic acid esters in house dust and home characteristics/living habits in a rural region of Taiwan.

Phthalic acid esters (PAEs) commonly used as plasticizers are distributed ubiquitously in the living environment. We conducted a field study to examine the associations between PAE residue in dust and home characteristics/living habits in 47 rural homes in Taiwan. A questionnaire regarding home characteristics/living habits and composite sampling of house dust were conducted in each participating home. Five PAEs were selected for analysis on gas chromatography-mass spectrometry with the limits of quantification being 0.5 ng/g or lower. Uni- and multivariate linear regression analyses were performed for examining the associations. The five PAEs were prevalently detected from the samples, and the concentrations were below 1000 ng/g; di(2-ethylhexyl) phthalate (DEHP) was the most frequently detected PAE (85%), whereas di-isobutyl phthalate (DiBP) appeared to the most abundant congener with the maximum concentration of 807.65 ng/g. Floor cleaning frequency and use of detergents for floor cleaning were significantly associated with DEHP in dust (P < 0.05), suggesting additives of plasticizers in detergent products. The factors of plastic wraps in storage and use of disposable cups were both significantly related to DiBP (P < 0.01), which could be extensively used in food packaging products. We confirmed that several home characteristics/living habits were related to certain PAE residue in dust.

Association between Pesticides in House Dust and Residential Proximity to Farmland in a Rural Region of Taiwan.

Pesticide drift was reported in many international studies, but rarely studied in Taiwan. We conducted a study in a rural region of Taiwan to examine the associations between pesticides in house dust and nearby agricultural areas using geographic information system (GIS). A questionnaire regarding home characteristics and pesticide use, and indoor and outdoor dust samples were collected from 47 rural homes. Dust samples were analyzed for six pesticides, and agricultural land data for GIS analysis were retrieved from a national website. All but prallethrin were frequently detected from indoor dust samples (>50%), and the maximum concentrations were all below 1000 ng/g. Detection frequencies and concentrations of pesticides in outdoor dust samples were even lower than that in indoor dust samples. Only "work involving pesticides" in the questionnaire was significantly associated with four pesticides in house dust (p < 0.05). Carbofuran and tetramethrin in house dust were significantly correlated with rice cultivation area at certain buffer distances (ρ > 0.33, p < 0.05), and chlorpyrifos was found to be associated with abandoned cultivation area, suggesting the occurrence of pesticide drift. Despite the low levels of pesticides in house dust, residents in the rural region should be cautious of pesticide drift from nearby active or abandoned farmlands.

Optimization of a Portable Adenosine Triphosphate Bioluminescence Assay Coupled with a Receiver Operating Characteristic Model to Assess Bioaerosol Concentrations on Site.

Rapid monitoring of the microbial content in indoor air is an important issue. In this study, we develop a method for applying a Coriolis sampler coupled with a portable ATP luminometer for characterization of the collection efficiency of bioaerosol samplers and then test this approach in field applications. The biological collection efficiencies of the Coriolis sampler and a BioSampler for collecting four different types of bioaerosols, including Escherichia coli, Staphylococcus aureus, Candida famata and endospores of Bacillus subtilis, were compared in a chamber study. The results showed that the ATP assay may indicate the four microbes' viability, and that their defined viabilities were positively correlated with their culturability. In addition, the optimal sampling conditions of the Coriolis sampler were a 200 L/min flow rate and a sampling time of 30 min. Under these conditions, there was no significant difference in sampling performance between the BioSampler and Coriolis sampler. In field applications, the best ATP benchmark that corresponded to culturable levels of < 500 CFU/m3 was 287 RLUs (sensitivity: 100%; specificity: 80%) for bacteria and 370 RLUs (sensitivity: 79%; specificity: 82%) for fungi according to receiver operating characteristic curve analysis. Consequently, an ATP criterion is recommended for indicating whether the corresponding airborne culturable concentrations of microbes meet those of published guidelines.

Relationship between Organophosphate and Pyrethroid Insecticides in Blood and Their Metabolites in Urine: A Pilot Study.

We conducted a pilot study to examine the relationship between organophosphate (OP) and pyrethroid (PYR) insecticides in blood and their metabolites in urine. A total of 30 pregnant women were enrolled in the study, and blood and urine was sampled from each subject during a regular clinic visit. Two OP and nine PYR insecticides were selected for blood sample analysis, while six OP and five PYR metabolites were analyzed for urine specimens. Both types of samples were processed and analyzed on gas chromatography-mass spectrometry. For OPs in blood, chlorpyrifos had a higher mean concentration (73.33 µg/L) than terbufos. For PYRs in blood, cypermethrin and imiprothrin were the most frequently detected species with the highest mean concentrations (151.25 and 141.25 µg/L). The concentrations of PYRs appeared to be higher than that of OPs, and the most frequently detected PYRs were commonly used in domestic products, suggesting that the exposure could mostly originate from use of domestic insecticides. The correlation between insecticides in blood and their metabolites in urine was significantly high (r = 0.795 for OPs and 0.882 for PYRs, p < 0.001), indicating routine exposure at a steady state. Residents should be cautious with domestic use of insecticide products to lower their exposure.

Analysis of Leafy Vegetable Nitrate Using a Modified Spectrometric Method.

A quick and cost-effective method to analyze leafy vegetable nitrate on spectrometry was developed and compared with a standard method using high performance liquid chromatography (HPLC). This method was designed to use ion-exchange solid phase extraction (SPE) cartridges in reducing interference from organic matrices to meet the criterion of an existing method for analyzing nitrate in wastewater. Nine vegetables (bok coy, cabbage, celery, Chinese cabbage, Chinese kale, lettuce, mustard green, pak choi, and water spinach) were selected for the method testing with three replicates being conducted for each vegetable. The nitrate contents ranged from 800 to 4,300 µg/g, with bok coy, celery, and pak choi being the highest. Data derived from spectrometry and HPLC were close to each other with most relative errors being within ±10% and were highly correlated with an R square value of 0.969. Stability testing and spike analysis resulted in a mean coefficient of variation lower than 6% and a mean recovery rate of 83.7%, suggesting reliability of the method. In addition, both the cost and time consumption for using this method were lower than the standard method using HPLC or ion chromatography, making this spectrometric method a good alternative for analysis of leafy vegetable nitrate.