A severe head louse (Pediculus humanus capitis) infestation: Timeline, pathogen detection, kdr-gene screening and morphological anomalies of lice.

Lice are assuming an increasing importance in forensic investigations, given their capacity to provide information about an individual's care. Head louse pediculosis is a frequent condition in school-age children and can be properly controlled using topical treatments combined with good personal hygiene. Prolonged and chronic infestations may result in more serious outcomes including severe iron deficiency anaemia. We conducted entomological and laboratory investigations of a head louse infestation in a 12-year-old girl who experienced severe anaemia and subsequent death. Numerous lice were found postmortem on the head, face and neck of the patient, as well as on bedding and clothing. Analysis of nits on individual hairs determined that the louse infestation had been present for at least 166 days. The lice had some morphological traits characteristic of body lice: the third antennal segment in some specimens was distinctly longer than wide, and the apices of some paratergal plates did not extend into intersegmental membranes, while other morphological features were characteristic of head or body lice. All lice were heterozygous for the T917I kdr genotype, a marker of permethrin resistance. Nineteen (79.2%, 95%CI 59.5%-90.8%) louse DNA samples tested TaqMan positive for Acinetobacter (Moraxellales; Moraxellaceae) sp. Available information and laboratory findings are further discussed regarding their possible contribution to the negative outcome of this case. We stress the impact head louse pediculosis can have on children with limited parental attention, and how severe head louse infestation may serve as warning sign of neglect, and other high-risk situations.

Improved efficiency of Sedum lineare (Crassulaceae) in remediation of arsenic-contaminated soil by phosphate-dissolving strain P-1 in association with phosphate rock.

The selection of appropriate plants and growth strategies is a key factor in improving the efficiency and universal applicability of phytoremediation. Sedum lineare grows rapidly and tolerates multiple adversities. The effects of inoculation of Acinetobacter sp. phosphate solubilizing bacteria P-1 and application of phosphate rock (PR) as additives on the remediation efficiency of As-contaminated soil by S. lineare were investigated. Compared with the control, both the single treatment and the combination of inoculation with strain P-1 and application of PR improved the biomass by 30.7-395.5%, chlorophyll content by 48.1-134.8%, total protein content by 12.5-92.4% and total As accumulation by 45.1-177.5%, and reduced the As-induced oxidative damage. Inoculation with strain P-1 increased the activities of superoxide dismutases and catalases of S. lineare under As stress, decreased the accumulation of reactive oxygen species in plant tissues and promoted the accumulation of As in roots. In contrast, simultaneous application of PR decreased As concentration in S. lineare tissues, attenuated As-induced lipid peroxidation and improved As transport to shoots. In addition, the combined application showed the best performance in improving resistance and biomass, which significantly increased root length by 149.1%, shoot length by 33%, fresh weight by 395.5% and total arsenic accumulation by 159.2%, but decreased the malondialdehyde content by 89.1%. Our results indicate that the combined application of strain P-1 and PR with S. lineare is a promising bioremediation strategy to accelerate phytoremediation of As-contaminated soils.

A novel enzyme synthesized by Acinetobacter sp. SM04 is responsible for zearalenone biodegradation.

Zearalenone (ZEA), a nonsteroidal estrogenic mycotoxin produced by multiple Fusarium species, contaminates cereals and threatens the health of both humans and animals by inducing hepatotoxicity, immunotoxicity, and genotoxicity. A new alkali tolerant enzyme named Ase, capable of degrading ZEA without H2O2, was derived from Acinetobacter sp. SM04 in this study. The Ase gene shares 97% sequence identity with hypothetical proteins from Acinetobacter pittii strain WCHAP 100004 and YMC 2010/8/T346 and Acinetobacter calcoaceticus PHEA-2, respectively. Based on the Acinetobacter genus database, the gene encoding Ase was cloned and extracellularly expressed in Escherichia coli BL21. After degrading 88.4% of ZEA (20 µg/mL), it was confirmed through MCF-7 cell proliferation assays that Ase can transform ZEA into a nonestrogenic toxic metabolite. Recombinant Ase (molecular weight: 28 kDa), produced by E. coli BL21/pET32a(+)-His-Ase, was identified as an oxygen-utilizing and cytochrome-related enzyme with optimal activity at 60 °C and pH 9.0.

A comprehensive study on bisphenol A degradation by newly isolated strains Acinetobacter sp. K1MN and Pseudomonas sp. BG12.

Bisphenol A (BPA) is an endocrine disrupting chemical. Its extensive use has led to the wide occurrence of BPA in various environmental ecosystems, at levels that may cause negative effects to the ecosystem and public health. Although there are many bacteria able to BPA utilization, only a few of them have a strong capacity for its biodegradation. Therefore, it is important to search for new bacteria strains, investigate their BPA biodegradation ability and potential effect of pH and other organic compounds on the process. These tasks have become the object of the present study. The results of our research show that for the newly isolated strains Acinetobacter sp. K1MN and Pseudomonas sp. BG12 after 15 days, with an initial BPA concentration of 100 mg L- 1, the highest BPA removal was achieved at pH 8, while sodium glutamate as a biostimulant best accelerated BPA degradation. Kinetic data for BPA biodegradation by both strains best fitted the Monod model. The specific degradation rate and the half saturation constant were estimated respectively as 8.75 mg L- 1 day- 1 and 111.27 mg L- 1 for Acinetobacter sp. K1MN, and 8.6 mg L- 1 day- 1 and 135.79 mg L- 1 for Pseudomonas sp. BG12. The half-maximal effective concentration (EC50) of BPA for Acinetobacter sp. K1MN was 120 mg L- 1 and for Pseudomonas sp. BG12 it was 123 mg L- 1. The toxicity bioassay (Microtox test) showed that elimination of BPA by both strains is accompanied by reduction of its toxic effect. The ability of tested strains to degrade BPA combined with their high resistance to this xenobiotic indicates that Acinetobacter sp. K1MN and Pseudomonas sp. BG12 are potential tools for BPA removal during wastewater treatment plant.

Protocatechuic acid production from lignin-associated phenolics.

Biobased chemicals are gaining popularity and market in attempts to mitigate the deteriorating environmental and sustainability issues. Components of renewable agricultural and forest biomass residues are projected to serve as abundant precursors to synthesis of expanding range of products. Agroindustrial wastes comprises of several phenolic compounds associated with lignin via ether linkages such as ferulic acid, p-coumaric, syringic acid and vanillin. These aromatic chemicals have myriad industrial applications. In this study, p-coumaric acid and ferulic acid were found to be two major components in corn bran derived lignin hydrolysate. Engineered Pseudomonas putida KT2440 was constructed and found to convert p-coumaric acid and vanillic acid to protocatechuic acid in >90% and >50% yields, respectively. Engineering the strain included deletion of the gene encoding protocatechuate 3,4-dioxygenase, and overexpression of vanillate-O-demethylase gene from Acinetobacter sp. ADP1.

The performance and mechanism of simultaneous removal of fluoride, calcium, and nitrate by calcium precipitating strain Acinetobacter sp. H12.

A calcium precipitating and denitrifying bacterium H12 was used to investigate the F- removal performance and mechanism. The results showed that the strain H12 reduced 85.24% (0.036 mg·L-1·h-1) of F-, 62.43% (0.94 mg·L-1·h-1) of Ca2+, and approximately 100% of NO3- over 120 h in continuous determination experiments. The response surface methodology analysis demonstrated that the maximum removal efficiency of F- was 88.98% (0.062 mg·L-1·h-1) within 72 h under the following conditions: the initial Ca2+ concentration of 250.00 mg·L-1, pH of 7.50, and the initial C4H4Na2O4·6H2O concentration of 800.00 mg·L-1. The scanning electron microscopy images, the X-ray photoelectron spectroscopy, and X-ray diffraction results suggested the following removal mechanism of F-: (1) the bacteria, as the nucleation site, were encapsulated by bioprecipitation to form biological crystal seeds; (2) Biological crystal seeds adsorbed F- to form Ca5(PO4)3F and CaF2; (3) Under the induction of bacteria, calcium, fluoride and phosphate coprecipitated to form Ca5(PO4)3F and CaF2. In addition, the gas chromatography data indicated that F- had little or no effect on the gas composition during denitrification, and the fluorescence spectroscopy analysis also proved that the extracellular polymeric substance (protein) is the site of bioprecipitation nucleation.

Acinetobacter sp. mediated synthesis of AgNPs, its optimization, characterization and synergistic antifungal activity against C. albicans.

AIMS: To synthesize silver nanoparticles (AgNPs) with cell free extract of Acinetobacter sp. and evaluate antifungal activity against planktonic and biofilm of Candida. Also, to study mechanism of antifungal action of AgNPs. METHODS AND RESULT: Acinetobacter spp were screened for synthesis of AgNPs. Physio-chemical parameters were optimized to obtained monodispersed nanoparticles. Optimized nanoparticles were characterized using spectroscopic, microscopic and diffraction techniques. Antifungal and biofilm disruption activity of AgNPs (10 ± 5 nm) were investigated against C. albicans. Mechanism of antifungal activity of nanosilver was deduced by growth curve, reactive oxygen species generation, thiol interaction and microscopic analysis. Acinetobacter sp. GWRFH 45 gave maximum synthesis of AgNPs. At optimized condition monodispersed, spherical nanoparticles were obtained which were crystalline with negative surface charge. AgNPs exhibited antifungal activity against planktonic cells and biofilm of Candida. AgNPs showed synergistic effect with amphotericin B as well as fluconazole against biofilm disruption. AgNPs were found to affect growth of Candida, generate reactive oxygen species and disrupt cellular morphology. CONCLUSIONS: Cell free extract of A. calcoaceticus GWRFH 45 has ability to synthesize AgNPs. AgNPs alone and in combination with drugs have potential to inhibit C. albicans. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of bacteriogenic AgNPs used in combination with antifungal drugs against Candida.

Optimization, purification, and characterization of hydroxylamine oxidoreductase from Acinetobacter sp. Y1.

Hydroxylamine oxidoreductase (HAO) is a key enzyme involved in ammonium removal pathway. To further study the enzyme, HAO was purified from heterotrophic nitrifier Acinetobacter sp. Y1 and its property was investigated. Results of single-factor experiments showed that the optimal carbon source, nitrogen source, and C/N ratio were trisodium citrate, ammonium sulfate, and 14, respectively, with incubation time of 16 H. DEAE SefinoseTM FF anion-exchange chromatography was used to purify HAO, followed by SefinoseTM CL-6B gel filtration chromatography. SDS-PAGE revealed that a 47 kDa enzyme was purified successfully, with a purification fold of 7.32 and a recovery rate of 19.40%. The optimized enzyme activity of purified HAO was tested at pH 8.0 and 30 °C. The results showed that the activity was increased by 43.78% and 25.64% in the presence of 1 mM Fe2+ and Fe3+ , respectively. HAO activity was increased with the increase of Na+ and K+ , Mn2+ , Zn2+ , Cu2+ , Ca2+ , Ba2+ inhibited the HAO activity at three concentrations. In addition, HAO activity was activated by ethylenediaminetetraacetic acid at 0.4 mM, and a negative effect arose as the dose increased. The purified enzyme from Y1 is different from other reported HAOs. Further study should be conducted to investigate the enzyme.

Novel Antimicrobial Indolepyrazines A and B from the Marine-Associated Acinetobacter sp. ZZ1275.

Two new alkaloids indolepyrazines A (1) and B (2) were isolated from the marine-derived Acinetobacter sp. ZZ1275. Their structures were elucidated through extensive nuclear magnetic resonance (NMR) spectroscopic analyses, high resolution electrospray ionization mass spectroscopy (HRESIMS) data, and electronic circular dichroism (ECD) calculation. Indolepyrazine A represents the first example of alkaloids with an indole-pyrazine-oxindole skeleton. Both 1 and 2 showed antimicrobial activities against methicillin-resistant Staphylococcus aureus, Escherichia coli, and Candida albicans with minimum inhibitory concentration (MIC) values of 12 µg/mL, 8⁻10 µg/mL, and 12⁻14 µg/mL, respectively.

Biodegradation and metabolic pathway of sulfamethoxazole by a novel strain Acinetobacter sp.

In this study, a novel strain capable of degrading sulfamethoxazole (SMX) was isolated and identified as Acinetobacter sp. The effect of influencing factors, such as initial SMX concentration (5-240 mg/L), temperature (15-35 °C), and pH (5-7), on SMX degradation was investigated. The results showed that when the initial SMX concentration was in the range of 5-240, the removal efficiency was 100%. The optimal condition for SMX biodegradation and microbial growth was determined to be 25 °C and pH = 7.0 in terms of the removal efficiencies of SMX and total organic carbon (TOC). Four metabolite compounds were detected during the process of SMX biodegradation, and the degradation pathways were tentatively proposed. In summary, Acinetobacter sp. was highly efficient in mineralizing SMX, which has the potential to be used for degrading SMX in water and wastewater.

Biobegradation and metabolic mechanism of cyprodinil by strain Acinetobacter sp. from a contaminated-agricultural soil in China.

Using sequential soil and liquid culture enrichments with cyprodinil as the sole carbon source, a Gram-negative cyprodinil-degrader from cyprodinil-polluted agricultural soil was isolated. The sequencing analysis of 16 S rRNA indicated that the strain showed 99% homology to Acinetobacter sp. The strain could effectively degrade cyprodinil at the neutral condition. At the initial concentrations of 10, 20, 50, 100, 150 and 200 mg L-1 in minimal medium, cyprodinil was degraded by 10, 20, 49.3, 64.2, 57 and 24 mg L-1 within 14 days, respectively. Two metabolites (4-cyclopropyl-6-methyl-2-pyrimidpyridine amine and monohydroxylated para-substitution) were identified using high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (HPLC-QTOF-MS/MS). A biodegradation pathway involving imines hydrolysis and monohydroxyl substitution on benzene ring was proposed on basis of the identified metabolites. Acinetobacter sp. would have a potential application in bioremediation of cyprodinil-contaminated soil, and the strain might have important implications in detoxification and bioremediation of pyrimidine analogues.

High phenol degradation capacity of a newly characterized Acinetobacter sp. SA01: Bacterial cell viability and membrane impairment in respect to the phenol toxicity.

An efficient phenol-degrading bacterial strain, belonging to Acinetobacter genus, was isolated and selected to study the impact of different environmentally relevant phenol concentrations on the degradation process. The bacterial isolate, labeled as Acinetobacter sp. SA01 was able to degrade the maximum phenol concentration of 1 g/l during 60 h at optimum condition of pH 7, 30 °C and 180 rpm. Aeration and initial cell density, the two important factors, were carefully examined in the optimal growth conditions. The results showed that these two variables related proportionally with phenol degradation rate. Further investigations showed no effect of inoculum size on the enhancement of degradation of phenol at over 1 g/l. Flow cytometry (FCM) study was performed to find out the relationship between phenol-induced damages and phenol degradation process. Single staining using propidium iodide (PI) showed increased cell membrane permeability with an increase of phenol concentration, while single staining with carboxyfluorescein diacetate (cFDA) demonstrated a considerable reduction in esterase activity of the cells treated with phenol at more than 1 g/l. A detailed investigation of cellular viability using concurrent double staining of cFDA/PI revealed that the cell death increases in cells exposed to phenol at more than 1 g/l. The rate of cell death was low but noticeable in the presence of phenol concentration of 2 g/l, over time. Phenol at concentrations of 3 and 4 g/l caused strong toxicity in living cells of Acinetobacter sp. SA01. The plate count method and microscopy analysis of the cells treated with phenol at 1.5 and 2 g/l confirmed an apparent reduction in cell number over time. It was assumed that the phenol concentrations higher than 1 g/l have destructive effects on membrane integrity of Acinetobacter sp. SA01. Our results also revealed that the toxicity did not reduce by increasing initial cell density. Scanning electron microscopy (SEM) examination of bacterial cells revealed the surface morphological changes following exposure to phenol. The bacterial cells, with wizened appearance and wrinkled surface, were observed by exposing to phenol (1 g/l) at lag phase. A morphological change occurred in the mid-logarithmic phase as the bacterial cells demonstrated coccobacilli form as well as elongated filamentous shape. The wrinkled cell surface were totally disappeared in mid-stationary phase, suggesting that the complete degradation of phenol relieve the stress and direct bacterial cells toward possessing smoother cell membrane.

Stress responses of Acinetobacter strain Y during phenol degradation.

Quantification of gene expression of Acinetobacter strain Y under 1000 mg/l of phenol was investigated using qPCR and proteomic analyses. The results show that Acinetobacter strain Y utilized 100 % of phenol within 18 h of exposure. The results of qPCR and proteomic analyses demonstrate a sequential expression of phenol-degrading genes of Acinetobacter strain Y via the ortho-pathway followed by the ß-ketoadipate pathway. Many stress-responsive proteins such as chaperones, chaperonins, porins and the enzymes involved in the signal transduction pathway were upregulated especially in the early stage. The stressed bacteria produced more ABC-type transporters, membrane receptors and efflux pumps to mitigate the impacts of phenol stress. The functions of TCA/glyoxylate cycle and oxidative phosphorylation processes were negatively affected. Many enzymes in the gluconeogenesis pathway were upregulated. This study demonstrates bacterial strategies of Acinetobacter strain Y via the energy saving mechanisms and the coordinated control between carbon (C)- and nitrogen (N)-limitations in coping with the stress by scavenging the reactive oxygen species.

Quorum sensing molecules production by nosocomial and soil isolates Acinetobacter baumannii.

Acinetobacter species remain alive in hospitals on various surfaces, both dry and moist, forming an important source of hospital infections. These bacteria are naturally resistant to many antibiotic classes. Although the role of the quorum sensing system in regulating the virulence factors of Acinetobacter species has not been fully elucidated, it has been reported that they play a role in bacterial biofilm formation. The biofilm formation helps them to survive under unfavorable growth conditions and antimicrobial treatments. It is based on the accumulation of bacterial communication signal molecules in the area. In this study, we compared the bacterial signal molecules of 50 nosocomial Acinetobacter baumannii strain and 20 A. baumannii strain isolated from soil. The signal molecules were detected by the biosensor bacteria (Chromobacterium violaceum 026, Agrobacterium tumefaciens A136, and Agrobacterium tumefaciens NTL1) and their separation was determined by thin-layer chromatography. As a result, it has been found that soil-borne isolates can produce 3-oxo-C8-AHL and C8-AHL, whereas nosocomial-derived isolates can produce long-chain signals such as C10-AHL, C12-AHL, C14-AHL and C16-AHL. According to these results, it is possible to understand that these signal molecules are found in the infection caused by A. baumannii. The inhibition of this signaling molecules in a communication could use to prevent multiple antibiotic resistance of these bacteria.

PCR Assay Based on the gyrB Gene for Rapid Identification of Acinetobacter baumannii-calcoaceticus Complex at Specie Level.

BACKGROUND: The genus Acinetobacter sp. comprises more than 50 species, and four are closely related and difficult to be distinguished by either phenotypic or genotypic methods: the Acinetobacter calcoaceticus-baumannii complex (ABC). The correct identification at species level is necessary mainly due to the epidemiological aspects. METHODS: We evaluated a multiplex PCR for gyrB gene to identify the species of the ABC using the sequencing of the ITS 16S-23S fragment as a gold standard. Isolates identified as Acinetobacter calcoaceticus-baumannii from three hospitals at southern Brazil in 2011 were included in this study. RESULTS: A total of 117 isolates were obtained and 106 (90.6%) were confirmed as A. baumannii, 6 (5.1%) as A. nosocomialis and 4 (3.4%) as A. pittii by PCR for gyrB gene. Only one isolate did not present a product of the PCR for the gyrB gene; this isolate was identified as Acinetobacter genospecie 10 by sequencing of ITS. We also noted that the non-A. baumannii isolates were recovered from respiratory tract (8/72.7%), blood (2/18.2%) and urine (1/9.1%), suggesting that these species can cause serious infection. CONCLUSION: These findings evidenced that the multiplex PCR of the gyrB is a feasible and simple method to identify isolates of the ABC at the species level.

Medium chain length polyhydroxyalkanoates consisting primarily of unsaturated 3-hydroxy-5-cis-dodecanoate synthesized by newly isolated bacteria using crude glycerol.

BACKGROUND: Our study aimed to search for novel bacteria capable of producing polyhydroxyalkanoates (PHAs) using crude glycerol residue obtained from biodiesel production in which used cooking oils were the substrates. RESULTS: Newly isolated bacteria from soils in Thailand were screened for the efficient production of PHAs from crude glycerol. The bacterial strains were cultivated on glucose, refined glycerol, crude glycerol, or various cooking oils (canola oil, palm oil, soybean oil, sunflower oil, corn oil, grape seed oil, olive oil, rice bran oil, camellia seed oil) for growth and PHA production. The effects of the total organic carbon (TOC) concentration and the mole ratio of carbon to nitrogen were investigated in batch cultivation. (1)H NMR, two dimensional-(1)H-correlation spectroscopy (2D-(1)H-COSY) and (13)C NMR analyses confirmed four bacterial strains were capable of producing medium-chain-length PHAs (mcl-PHAs), consisting of 3-hydroxyoctanoate (3HO) and 3-hydroxy-5-cis-dodecanoate (3H5DD), from crude glycerol. On the basis of phenotypic features and genotypic investigations, the bacterial strains were assigned as: ASC1, Acinetobacter genus (94.9% similarity); ASC2, Pseudomonas genus (99.2% similarity); ASC3, Enterobacter genus (99.2% similarity); ASC4, Bacillus genus (98.4% similarity). The highest amount of mcl-PHAs, 17.5 ± 0.8 g/L (content 61.8 ± 3.3% wt), with 3HO (14.7 ± 2.2 mol %), 3H5DD (85.3 ± 2.2 mol%), and a total biomass of 32.3 ± 0.3 g/L, was obtained from Pseudomonas sp. ASC2 in batch cultivation after 36 h. The mcl-PHAs recovered had a number-average molecular weight (M N) of 3.6 × 10(4) Da. Homopolymeric 3H5DD was obtained when the cultivation time was prolonged to 96 h. CONCLUSIONS: Novel PHA-producing strains were isolated and identified. These bacterial strains are able to produce mcl-PHAs from crude glycerol. The mcl-PHAs produced contained a high percentage of 3H5DD, which suggests their future application as softeners mixed with other biomaterials. The unsaturated side chain of 3H5DD monomers containing double bounds offers additional potential for improving the properties of the mcl-PHAs or extending their applications to the food industry.

A metallo-keratinase from a newly isolated Acinetobacter sp. R-1 with low collagenase activity and its biotechnological application potential in leather industry.

Microbial keratinase is a well-recognized enzyme that can specifically degrade insoluble keratins. A keratinase-producing bacterium was isolated from a duck ranch soil and identified as Acinetobacter sp. R-1 based on the biochemical characteristics and 16S rDNA gene sequencing. It showed high keratinase activity and low collagenase activity. The keratinase was purified to electrophoretic homogeneity with 6.69% recovery, 2.68-fold purification and an estimated molecular weight of 25 kDa. Additionally, the keratinase showed optimal activity at 50 °C and pH11. Keratinase activity of Acinetobacter sp. significantly increased in the presence of Li(+), Na(+), and Ca(2+), while it was completely inhibited by EDTA, indicating it was a metallo-keratinase. Moreover, the crude keratinase from Acinetobacter sp. R-1 could thoroughly depilate goat skin and simultaneously modify the wool surface, which indicated its applicable potential in leather and textile industries.

Simultaneous enhancement of phenolic compound degradations by Acinetobacter strain V2 via a step-wise continuous acclimation process.

Phenol degradation enhancement of Acinetobacter strain V2 by a step-wise continuous acclimation process was investigated. At the end of 8 months, three stable adapted strains, designated as R, G, and Y, were developed with the sub-lethal concentration of phenol at 800, 1100, and 1400 mg/L, respectively, from 400 mg/L of V2 parent strain. All strains degraded phenol at their sub-lethal level within 24 h, their growth rate increased as the acclimation process continued and retained their degradation properties even after storing at -80 °C for more than 3 years. All adapted strains appeared coccoid with an ungranulated surface under electron microscope compared to typical rod-shaped parental strain V2 . The adapted Y strain also possessed superior degradation ability against aniline, benzoate, and toluene. This study demonstrated the use of long term acclimation process to develop efficient and better pollutant degrading bacterial strains with potentials in industrial and environmental bioremediation.

Consequences of antagonistic interactions between endophytic fungus and bacterium on plant growth and defense responses in Atractylodes lancea.

Many studies have examined pair-wise interactions between plants and endophytes, while overlooking the interplays among multiple endosymbionts and their combined impacts on hosts. In this study, Atractylodes lancea plantlets were inoculated with endophytic fungus Acremonium strictum AL16, or endophytic bacterium Acinetobacter sp., or both, to investigate the impacts of the three-way symbiosis on the host. Our results showed that defense-related responses of the co-inoculated plantlets were delayed and weakened relative to plantlets with single inoculants, but no detrimental effects on phyto-physiology (growth, photosynthesis) were observed after combined inoculations. Quantitative PCR analysis verified a decrease in AL16 colonization density within plants after co-inoculation with the endobacteria. An in vitro assay was then performed to elucidate the suppressed plant defense responses and reduced fungal colonization by dual inoculation. The data showed that the presence of Acinetobacter sp. reduced AL16 colony diameter and spore germination rate without negatively affecting fungal morphology. Additionally, direct hyphal attachment of the bacterium to AL16 in vitro was visualized by scanning electronic microscopy. Therefore, we propose that a balanced and compatible symbiosis might require constraints conferred by the antagonistic endophyte Acinetobacter sp. on the fungus AL16 in the tripartite endophytic bacterium-fungus-plant system.

Biodegradation of type II pyrethroids and major degraded products by a newly isolated Acinetobacter sp. strain JN8.

A Gram-negative aerobic bacterium, designated as JN8, was isolated from activated sludge and soil in a pesticides factory in China. It was found that JN8 had a high capacity for degrading a broad range of type II pyrethroids and utilizing these pyrethroids as the sole carbon source for cell growth. The degradation rates of a 100 mg·L(-1) concentration of ß-cypermethrin, cypermethrin, fenpropathrin, fenvalerate, and deltamethrin by JN8 in mineral salt medium were 74.1%, 64.9%, 57.9%, 48.1% and 34.9%, respectively. Strain JN8 was identified as a species of Acinetobacter based on its biochemical properties and 16S rRNA sequence analysis. ß-Cypermethrin was degraded by JN8 through hydrolysis of the carboxylester linkage to form 3-phenoxybenzoic acid and 3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane carboxylic acid, both of which could be further degraded by JN8. JN8 is the first strain of an Acinetobacter species in which pyrethoid-degrading activity has been detected, and such a feature makes it a potential resource for disposal of waste and effluent from pyrethroid manufacturing facilities.