Genome sequences of four bacterial strains isolated from organofluorine enrichment cultures.

Because fluorinated organic compounds are broadly used and highly persistent, microbes isolated from wastewater may be able to degrade these contaminants. Here, we report the genome sequences of Flavobacterium sp. str. WV_118_3, Nocardioides sp. str. WV_118_6, Ochrobactrum anthropi str. WV_118_8, and Sphingomonas sp. str. VL_57B, isolated from wastewater.

[Determination of fluoroacetic acid in human blood and urine by accelerated solvent extraction-ion chromatography-mass spectrometry].

Fluoroacetic acid is a highly polar poison used for rodent control. When ingested by the human body, it seriously damages nerve cells and heart tissues and even causes death by cardiac arrest or respiratory failure. Common detection methods for fluoroacetic acid include gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry, both of which require complex pretreatment methods, such as derivatization. In this study, a method to determine fluoroacetic acid in human blood and urine based on accelerated solvent extraction-ion chromatography-mass spectrometry (ASE-IC-MS) was established. Two pretreatment methods, namely, acetonitrile precipitation and accelerated solvent extraction, were compared. Furthermore, the effects of different extraction conditions, such as the extraction time, extraction temperature, and number of cycles, were investigated. The most suitable chromatographic separation conditions, such as the chromatographic column, column temperature, and elution procedure, were determined, and the MS conditions, such as the collision energy (CE) and declustering potential (DP) of the ion pairs of the target compound, were investigated. Based on the experimental results, the optimal pretreatment methods and detection conditions were obtained, and reliable data were collected. Deionized water was used as the extraction solvent, and blood and urine samples were processed by accelerated solvent extractor. The supernatant was sequentially collected via centrifugal ultrafiltration and 0.22 µm membrane filtration, diluted 50 times, and then injected into the chromatographic column for detection. An Ion Pac AS20 IC column was used for isocratic elution with 15.0 mmol/L KOH solution as the eluent. The effluent was passed through a suppressor and into a triple quadrupole mass spectrometer, which was used to perform MS/MS (ESI-) in multiple reaction monitoring (MRM) mode. The quantitative ion was m/z 77.0>57.0 when the CE and DP were -15.0 eV and -20.0 V, respectively. An external standard method was used for quantitative analysis. The results showed a good linear relationship for fluoroacetic acid in the range of 0.5-500.0 µg/L (r>0.999), with limits of detection (LOD) and quantification (LOQ) of 0.14 and 0.47 µg/L, respectively. The recoveries of fluoroacetic acid in blood and urine were 93.4%-95.8% and 96.2%-98.4%, respectively. The intra-day RSDs for blood and urine were 0.8%-1.6% and 0.2%-1.0%, respectively, while the inter-day RSDs were 2.3%-3.8% and 3.9%-6.9%, respectively. Further investigation revealed that the matrix effects of this method in blood and urine, at -7.4% and -3.0%, respectively, were fairly weak. The established method was successfully applied to detect fluoroacetic acid in human blood and urine obtained from a poisoning case, and the results obtained provided crucial clues that led to swift case resolution. The efficiency of the method was significantly higher than that of conventional detection methods. In conclusion, the developed method has high sensitivity and good repeatability and is suitable for the rapid detection of fluoroacetic acid in human blood and urine. Moreover, because this method does not require derivatization, it is simple and efficient.

Fluoroacetate distribution, response to fluoridation, and synthesis in juvenile Gastrolobium bilobum plants.

Like angiosperms from several other families, the leguminous shrub Gastrolobium bilobum R.Br. produces and accumulates fluoroacetate, indicating that it performs the difficult chemistry needed to make a C-F bond. Bioinformatic analyses indicate that plants lack homologs of the only enzymes known to make a C-F bond, i.e., the Actinomycete flurorinases that form 5'-fluoro-5'-deoxyadenosine from S-adenosylmethionine and fluoride ion. To probe the origin of fluoroacetate in G. bilobum we first showed that fluoroacetate accumulates to millimolar levels in young leaves but not older leaves, stems or roots, that leaf fluoroacetate levels vary >20-fold between individual plants and are not markedly raised by sodium fluoride treatment. Young leaves were fed adenosine-13C-ribose, 13C-serine, or 13C-acetate to test plausible biosynthetic routes to fluoroacetate from S-adenosylmethionine, a C3-pyridoxal phosphate complex, or acetyl-CoA, respectively. Incorporation of 13C into expected metabolites confirmed that all three precursors were taken up and metabolized. Consistent with the bioinformatic evidence against an Actinomycete-type pathway, no adenosine-13C-ribose was converted to 13C-fluoroacetate; nor was the characteristic 4-fluorothreonine product of the Actinomycete pathway detected. Similarly, no 13C from acetate or serine was incorporated into fluoroacetate. While not fully excluding the hypothetical pathways that were tested, these negative labeling data imply that G. bilobum creates the C-F bond by an unprecedented biochemical reaction. Enzyme(s) that mediate such a reaction could be of great value in pharmaceutical and agrochemical manufacturing.

Plasma fluoroacetic acid concentrations: Symptoms, hematological, and biochemical characteristics in patients with fluoroacetic acid poisoning in the emergency department.

Fluoroacetic acid (FAcOH) was once a highly toxic rodenticide widely used in the world. In the past, studies on the toxicity of FAcOH have focused on animal experiments. The toxicity of FAcOH to humans and the changes of FAcOH in plasma have not been studied. Therefore, the present study aimed to describe the changes of plasma FAcOH concentrations, hematological, and biochemical characteristics in patients with FAcOH intoxication. According to clinical symptoms, 68 patients from the emergency department were divided into different groups: convulsion group, unconsciousness group, death group, and control groups. Plasma FAcOH concentrations, hematological, and biochemical parameters were investigated. Results demonstrated that patients in the convulsion group and the unconsciousness group had a significant increase (p < 0.01) in the level of neuron-specific enolase (NSE), creatine kinase MB (CKMB), glucose (GLU), and white blood cell count (WBC) and a significant decrease (p < 0.01) in serum potassium compared with the control group, respectively. Moreover, patients in the death group had a significant increase (p < 0.01) in the level of NSE, CKMB, N-terminal pro-brain natriuretic peptide, GLU, and WBC and a significant decrease (p < 0.01) in serum potassium and total calcium compared with the survival group. The concentrations of FAcOH in plasma in the convulsion group, the unconsciousness group, and the death group were 72.31 ± 42.29, 118.33 ± 55.41, and 163.78 ± 43.32 µg/mL, respectively. These changes and the plasma FAcOH concentrations may increase our understanding of the toxicity of FAcOH to humans and may help doctors to judge the clinical prognosis of patients with FAcOH intoxication.

Evidence for the Formation of Difluoroacetic Acid in Chlorofluorocarbon-Contaminated Ground Water.

The concentrations of difluoroacetic acid (DFA) and trifluoroacetic acid (TFA) in rainwater and surface water from Berlin, Germany resembled those reported for similar urban areas, and the TFA/DFA ratio in rainwater of 10:1 was in accordance with the literature. In contrast, nearby ground water historically contaminated with 1,1,2-trichloro-1,2,2-trifluoroethane (R113) displayed a TFA/DFA ratio of 1:3. This observation is discussed versus the inventory of microbial degradation products present in this ground water along with the parent R113 itself. A microbial transformation of chlorotrifluoroethylene (R1113) to DFA so far has not been reported for environmental media, and is suggested based on well-established mammalian metabolic pathways.

Evaluation of a novel thermo-alkaline Staphylococcus aureus lipase for application in detergent formulations.

An extracellular lipase of a newly isolated S. aureus strain ALA1 (SAL4) was purified from the optimized culture medium. The SAL4 specific activity determined at 60 °C and pH 12 by using olive oil emulsion or TC4, reached 7215 U/mg and 2484 U/mg, respectively. The 38 NH2-terminal amino acid sequence of the purified enzyme starting with two extra amino acid residues (LK) was similar to known staphylococcal lipase sequences. This novel lipase maintained almost 100% and 75% of its full activity in a pH range of 4.0-12 after a 24 h incubation or after 0.5 h treatment at 70 °C, respectively. Interestingly, SAL4 displayed appreciable stability toward oxidizing agents, anionic and non-ionic surfactants in addition to its compatibility with several commercial detergents. Overall, these interesting characteristics make this new lipase promising for its application in detergent industry.

Intoxicação por Mascagnia rigida (Malpighiaceae) em ovinos e caprinos / Poisoning by Mascagnia rigida (Malpighiaceae) in sheep and goats

Mascagnia rigida é a planta tóxica mais importante para bovinos na região Nordeste, causando morte súbita associada ao exercício. O presente trabalho teve como objetivos descrever três surtos de intoxicação por M. rigida em ovinos e um surto em caprinos no semi-árido da Paraíba, reproduzir experimentalmente a intoxicação em ovinos e caprinos, e comprovar a passagem do princípio ativo de M. rigida pelo colostro destes pequenos ruminantes. Os surtos ocorreram no início do período chuvoso, quando a planta brota antes do que outras forrageiras ou após o final desse período, quando após secarem algumas forrageiras, M. rigida permanece verde. Na reprodução experimental da intoxicação por M. rigida, doses de 10 e 20g/kg de peso animal, com as planta proveniente de duas regiões diferentes, foram letais para três caprinos e três ovinos. Um caprino que ingeriu 20g/kg da planta um ovino que ingeriu 10g/kg, se recuperaram. Dois ovinos e dois caprinos que ingeriram 5g/kg tiveram sinais discretos e se recuperaram. Tanto os casos experimentais quanto os espontâneos apresentaram ingurgitamento das veias jugulares, relutância em caminhar, decúbito externo abdominal, incoordenação, respiração ofegante, depressão, instabilidade e tremores musculares. A morte ocorreu após um curso clínico de alguns minutos a 27h40 min. As principais lesões foram edema pulmonar e vacuolização e necrose de células epiteliais dos túbulos renais. Para testar se o princípio ativo de M. rigida é eliminado pelo leite causando morte súbita nas crias foi realizado um experimento com duas cabras e cinco ovelhas que ingeriram, diariamente, 2g/kg de M. rigida, nos 15 dias anteriores ao parto. Uma ovelha que tinha gestação gemelar abortou depois de ter ingerido a planta por 10 dias. Os cordeiros das demais ovelhas mamaram normalmente o colostro sem aprestar sinais clínicos. O cabrito de uma cabra mamou o colostro e 5 minutos após morreu subitamente...(AU)

Mascagnia rigida is the most important toxic plant for cattle in the Northeastern region of Brazil, causing sudden death during exercise. The objectives of this research were to report three outbreaks of poisoning by M. rigida in sheep and one in goats in the semiarid of the state of Paraíba, to reproduce experimentally the disease, and to determine if the active principle of the plant is eliminated through the milk. The outbreaks occurred at beginning of the raining season, when the plant sprouts ahead other forages, or after the end of the raining season, when M. rigida stayed green and other forages had dried. In the experimental reproduction of the poisoning doses of 10 and 20g of M. rigida per kg body weight from two different regions were lethal to 3 goats and 3 sheep. One goat that ingested 20g/kg and a sheep that ingested 10g/kg recovered. Two sheep and two goats that ingested 5g/kg had mild clinical signs and recovered. Clinical signs of experimental and spontaneous cases were ingurgitation of the jugular veins, reluctance to move, sternal recumbence, incoordination, respiratory distress, depression, instability, and muscular tremors. Death occurred after a clinical manifestation period of about 4 min to 27h40min. The main lesions were pulmonary edema and vacuolization and necrosis of epithelial cells in some renal tubules. To test if the active principle of M. rigida causes sudden deaths in newborn lambs and kids, 2g/kg of the plant were given daily to two goats and five sheep in the 15 days previous to parturition. One sheep aborted two lambs, 5 days before parturition. The four lambs of the other four sheep ingested the colostrum without problems. The kid from one goat ingested the colostrum and died suddenly 5 minutes after. The kid from the other goat died immediately after parturition before ingestion of colostrum...(AU)

Mortes súbitas em bovinos causadas por Palicourea aeneofusca (Rubiaceae) e Mascagnia rigida (Malpighiaceae)na Zona da Mata Paraibana / Sudden deaths caused by Palicourea aeneofusca (Rubiaceae) and Mascagnia rigida (Malpighiaceae) in cattle in the Zona da Mata of Paraíba

Neste trabalho descrevem-se, em bovinos, um surto de intoxicação por Palicourea aeneofusca no município de Jacaraú e um surto de intoxicação por Mascagnia rigida, no município de Sapé. Ambos os surtos ocorreram na zona da Mata Paraibana, onde não havia, anteriormente, informação sobre intoxicação por plantas que causam mortes súbitas associadas ao exercício. A toxicidade de ambas as plantas frescas recém coletadas foi testada em coelhos, sendo a dose letal de 3g por kg de peso animal (g/kg) para P. aeneofusca e de 10g/kg para M. rigida. Na fazenda onde ocorreu o surto de intoxicação por P. aeneofusca foi encontrada, também M. rigida e o produtor informava que esta tinha causado, anteriormente, mortes de bovinos. M. rigida coletada nesta última fazenda foi tóxica na dose de 10g/kg da planta seca. Anteriormente a intoxicação por P. aeneofusca tinha sido diagnosticada na Zona da Mata e Agreste Pernambucano e leste da Bahia. Há também numerosos históricos da ocorrência desta intoxicação na Zona da Mata Alagoana, o que sugere sua ocorrência no litoral do nordeste, desde a Bahia até Paraíba. M. rigida é a planta tóxica mais conhecida e mais importante do nordeste ocorrendo no sertão e no agreste. Este trabalho comprova que, pelo menos na Paraíba, a intoxicação ocorre, também, na Zona da Mata.(AU)

This paper reports an outbreak of poisoning by Palicourea aeneofusca in cattle, in the municipality of Jacaraú, and another by Mascagnia rigida in the municipality of Sapé. Both outbreaks occurred in the Zona da Mata of Paraíba, where there are no previous reports of poisoning by toxic plants causing sudden death associated to exercise. The toxicity of both plants was tested in rabbits. The lethal dose was of 3g per kg body weight (g/kg) for fresh P. aeneofusca and 10g/kg for fresh Mascagnia rigida. On the farm where the poisoning by P. aeneofusca was observed, M. rigida also was found. The farmer reported that this plant also caused sudden death previously. M. rigida collected on that farm and given dried to rabbits caused death at the dose of 10gkg. The poisoning by P. aeneofusca in cattle had been previously reported in the Zona da Mata and Agreste of Pernambuco and east of Bahia. There are also numerous farmers' reports about the occurrence of the poisoning in the Zona da Mata of Alagoas, suggesting that it occurs in the whole coastal region, from Bahia to Paraíba. M. rigida is the most important toxic plant for cattle in the Brazilian semiarid. The results of this paper demonstrate that this plant poisoning also occurs in the tropical wet climate of the Zona da Mata.(AU)

Fluoroacetic Acid Is a Potent and Specific Inhibitor of Reproduction in the Nematode Caenorhabditis elegans.

Fluoroacetic acid is known to lead to inhibition of aconitase and block both the Krebs and glyoxylate cycles. In this study, we discovered it to be a potent and specific inhibitor of reproduction in a bioassay using the nematode Caenorhabditis elegans. Fluoroacetic acid added to the growth medium reduced reproduction in the second generation by 50% at concentrations 3,000 times lower than the concentrations that reduced 24-hour survival by 50%. Four concentrations (2, 4, 8, and 17 mM) of fluoroacetic acid were tested thoroughly. At the two lower concentrations, the survival rates were unaffected, and first-generation reproduction was greatly reduced but not completely eliminated. Survival was reduced at the higher concentrations. Malonate, which inhibits the Krebs cycle, and itaconate, which inhibits the glyoxylate cycle, were tested individually and in combination. The combination did not specifically inhibit reproduction, suggesting another mode of action for fluoroacetic acid. Fluoroacetic acid shows promise as a tool in studies requiring age synchrony.