Haemaphysalis flava ticks as a competent vector of severe fever with thrombocytopenia syndrome virus.

Severe fever with thrombocytopenia syndrome virus (SFTSV), a tick-borne Bunyavirus, causes an emerging hemorrhagic fever in humans with a high fatality in Asia. The tick vectors and hosts of SFTSV are not well studied. We evaluated SFTSV transmission in laboratory reared Haemaphysalis flava ticks. RT-PCR demonstrated that after acquisition feeding in SFTSV-infected rabbits, 10 % (4/40) engorged larvae, 25% (5/20) engorged nymphs, and 50% (5/10) engorged females of H. flava became SFTSV RNA positive; after engorged larvae and nymphs molted into nymphs and adults, respectively, 12.5% (3/24) newly molted nymphs and 20% (2/10) newly molted adults were SFTSV RNA positive. Among 30 engorged females that oviposited, 10% (3/30) clutches of eggs and 3.3% (1/30) colonies of larvae were RNA positive for SFTSV. RT-PCR also showed that 6 days after being infested with SFTSV-infected ticks, 100% (3/3) rabbits infested with larvae, 100% (2/2) rabbits infested with nymphs, and 100% (2/2) rabbits infested with adult ticks became SFTSV RNA positive. In conclusion, H. flava can acquire SFTSV from infected rabbits by feeding; there is transstadial and transovarial transmission of the virus and all three stages of H. flava can transmit SFTSV to rabbits by feeding. Thus, H. flava tick is an effective vector of SFTSV and may play a role in the transmission of SFTSV in wild animals and humans.

Pb(II)-mediated precipitate transformation promotes Cr(VI) immobilization by biogenic hydroxyapatite.

In this work, the mechanism of Pb(II)-mediated precipitation transformation to improve the removal of Cr(VI)-oxyanion on biogenic hydroxyapatite (BHAp) were investigated. The Pb(II)-preloading formed pyromorphite [Pb5(PO4)3Cl] precipitate on the BHAp surface (Pb@BHAp), thus causing an increase of 2.2 times in the uptake of Cr(VI) by Pb@BHAp at pH of 2.4. It was primarily due to the dissolution of Pb5(PO4)3Cl accompanied with the release of Pb(II), resulting in the rapid formation of crocoite (PbCrO4). Although the Ksp of Pb5(PO4)3Cl was approximately 23 orders of magnitude lower than that of PbCrO4, Pb(II)-mediated precipitation transformation could still occur. XRD and SEM-EDX analyses demonstrated that the process was a time-dependent that included rapid crystal precipitation in the initial 10 min and subsequent precipitate accumulation for several hours. The Pb(II) released from the dissolution of Pb5(PO4)3Cl was immediately immobilized by Cr(VI); therefore, it did not cause any retention risk of Pb(II) in the solution. Furthermore, a small quantity of Cr(VI) could be reduced to Cr(III) by BHAp, and Cr(III) could enter into the BHAp lattice for the exchange of Ca(II). This study provides a new insight into the resource utilization of Pb-bearing BHAp and a potential method for the successive removal of Pb(II) and Cr(VI).

Detection of Multiple Intracellular Bacterial Pathogens in Haemaphysalis flava Ticks Collected from Hedgehogs in Central China.

Tickborne intracellular bacterial pathogens including Anaplasma, Coxiella burnetti, Ehrlichia, and Rickettsia cause emerging infectious diseases worldwide. PCR was used to amplify the genes of these pathogens in Haemaphysalis flava ticks collected from hedgehogs in Central China. Among 125 samples including 20 egg batches, 24 engorged females, and 81 molted male and female adult ticks, the DNA sequences and phylogenetic analysis showed that the minimum infection rate of the ticks was 4% (5/125) for A. bovis, 3.2% (4/125) for C. burnetti, 9.6%, (12/125) for E. ewingii, and 5.6% for Rickettsia including R.japonica (3.2%, 4/125) and R. raoultii (2.4%, 3/125), respectively. The prevalence of these pathogens was significantly higher in dead engorged females (83.3%, 20/24) than in eggs (5%, 1/20) and molted ticks (8.6%, 7/81). Our study indicated that H. flava ticks could be infected with multiple species of tickborne pathogens including Anaplasma, C. burnetti, Ehrlichia, and Rickettsia in Central China, and the prevalence of these pathogens was reduced during transovarial and transstadial transmission in ticks, suggesting that ticks may not be real reservoirs but only vectors for these tickborne pathogens.

Pathogenic New World Relapsing Fever Borrelia in a Myotis Bat, Eastern China, 2015.

We identified Candidatus Borrelia fainii, a human pathogenic bacterium causing New World relapsing fever in a Myotis bat in eastern China. This finding expands knowledge about the geographic distribution of Borrelia spp. and the potential for infection with New World relapsing fever in China.

Occurrence and Genotyping of Coxiella burnetii in Hedgehogs in China.

Coxiella burnetii is the causative agent of query fever (Q fever), and distributes broadly in environment. Livestock are identified as main reservoirs, which may infect people through their contaminative urine, feces, milk, and birth products. Wild animals can also be the potential carriers and transmitters of C. burnetii. To understand the geographic distribution and host species of C. burnetii in China, we investigated the prevalence of C. burnetii in hedgehogs (Erinaceus amurensis) in Hubei Province. Hedgehogs were tested for C. burnetii with PCR targeting three genes (com1, rrs, and icd) followed by multispacer sequence typing (MST). We found that 12.2% (5/41) hedgehogs were PCR positive for C. burnetii. MST revealed presence of two novel genotypes and phylogenetic analysis revealed that the strains were similar to a group of isolates from chronic Q fever patients and mammals. This study showed that C. burnetii are highly prevalent in hedgehogs in Hubei Province in central China, suggesting that hedgehogs may play an important role in the ecology and transmission of C. burnetii to humans because it is captured and used as traditional medicine in China.

Detection of Leptospira interrogans in Hedgehogs from Central China.

Background:Leptospira is the causative agent of leptospirosis, a zoonotic disease of global importance. To have a better understanding on the host species of Leptospira, we investigated the prevalence of Leptospira species in hedgehogs in Central China. Materials and Methods: Hedgehogs were captured in Hubei Province, China in May and October, 2018. Total DNA was extracted from the kidney tissues of hedgehogs for determining the Leptospira species by PCR amplification of the rrs2, secY, and flaB genes with genus-specific primers. Results: PCR amplification indicated that the positive rate of hedgehogs to the rrs2, secY, and flaB genes were 19.5% (8/41), 12.2% (5/41), and 9.8% (4/41), respectively. The homology of the partial sequence of rrs2, secY, and flaB genes were 99.0-100% among the Leptospira strains from hedgehogs. Phylogenetic analysis revealed that Leptospira species detected in this study clustered together with Leptospira interrogans.Conclusions: We detected L. interrogans from hedgehogs in Central China, suggesting hedgehogs are the hosts of L. interrogans.

Lead competition alters the zinc adsorption mechanism on animal-derived biochar.

In this study, the adsorption behaviors and mechanisms of Pb(II) and Zn(II) by animal-derived biochar (ADB) in single and binary metal systems were comparatively investigated. ADB contains considerable amounts of Ca/P components and is mainly composed of hydroxyapatite (HAP), which plays an important role in the adsorption of Pb(II) and Zn(II). The maximum adsorption capacities of Pb(II) and Zn(II) on ADB were in the order of Zn(II)-single (3.23 mmol g-1) > Pb(II)-single (2.74 mmol g-1) ≈ Pb(II)-binary (2.71 mmol g-1) > Zn(II)-binary (2.31 mmol g-1). In the single metal system, approximately 99.9% of the adsorbed Pb(II) existed as Pb5(PO4)3Cl, while the dominant adsorption mechanism of Zn(II) was cation exchange, followed by precipitation, accounting for 78.0%-80.6% and 19.4%-21.5% of the adsorption capacity, respectively. These findings were verified by X-ray diffraction refinement, X-ray photoelectron spectroscopy, metal speciation modeling, and Ca(II) exchange experiment. In the binary metal system, the proportion and form of Pb(II) precipitate remained unchanged. However, the binding of Zn(II) to ADB was completely dependent on the cation exchange with Ca(II), and no remarkable Zn(II) precipitation was observed. Phosphate released from HAP preferentially precipitated with Pb(II) than with Zn(II) when they coexisted. Consequently, Pb(II) competition may alter the Zn(II) adsorption mechanism on ADB. Nonetheless, ADB could serve as an efficient biochar for the simultaneous immobilization of Pb(II) and Zn(II) via different mechanisms.

Hybrid ash/biochar biocomposites as soil amendments for the alleviation of cadmium accumulation by Oryza sativa L. in a contaminated paddy field.

A novel ash/biochar (A/B) biocomposite composed of 90% biomass bottom ash from agroforestry biomass direct-fired power plants, 5% animal-derived biochar from carcass pyrolysis, and 5% bentonite as an adhesive was amended in cadmium (Cd)-polluted paddy soil to alleviate cadmium accumulation by Oryza sativa L. Ash increased the soil pH and contributed exogenous available silicon. Biochar with high Ca/P components played an important role in soil cadmium immobilization. A 1-year field experiment with consecutive rice growing seasons (early and late rice) was conducted in Xiangtan, China, to examine the effects of A/B amendment in Cd-contaminated paddy soil. The A/B biocomposite was amended into soil through one-time addition at three application rates (1, 5, and 10 kg/m2). When A/B amendment was ≥5 kg/m2, the soil pH increased from 4.11 to more than 6. The available silicon content in the soil even increased by 22.9 times. For early rice soil, the CaCl2-extractable Cd(II) and toxicity characteristic leaching procedure (TCLP)-extractable Cd(II) decreased by 77.9%-96.1% and 52.4%-70.7%, respectively. A/B remarkably reduced Cd accumulation in rice organs, and this observation was related to A/B treatment rates. Ash and biochar contributed to the inhibition of Cd accumulation in rice organs and Cd translocation from roots to stems. The Cd concentrations in brown rice decreased to 0.11 and 0.12 mg/kg in early and late rice, respectively, and these values were lower than the national food safety standard limit value of China (0.2 mg/kg).

Mechanistic insights and multiple characterizations of cadmium binding to animal-derived biochar.

Cattle-derived biochar (CB), which is derived from industrial pyrolysis of cattle carcasses in harmless treatment plants, is a naturally occurring mineral form of carbonate-bearing hydroxyapatite (CHAP) with a small amount of elemental carbon. CB has 4.02% of carbonate content, which falls under the B-type substitution of CHAP. In this work, the Cd(II) sorption capacity of CB was determined to be 0.82 mmol/g, with 97.6% of the Cd(II) uptake contributing to CHAP and only 2.36% of the Cd(II) uptake contributing to the elemental carbon component. The calculation and linear combination fitting (LCF) of Cd L3-edge X-ray absorption near-edge structure (XANES) analysis indicated that the contributions of Cd(II) species to CB presented the following order: ion exchange (57.6%-61.0%) > precipitation (24.4%-29.9%) > surface complexation (12.5%-13.4%). The depth dependent X-ray photoelectron spectroscopy (XPS) showed the presence of ion exchange, which is accompanied by intraparticle diffusion. LCF of XANES and Rietveld analysis of X-ray diffraction (XRD) demonstrated that Cd(II) was precipitated in the form of Cd5H2(PO4)4·4H2O on the CB surface. Furthermore, the precipitate was directly observed and identified by scanning electron microscopy with energy-dispersive spectroscopy (SEM-EDS). Consequently, we revealed the intricate binding mechanism of Cd(II) to CHAP-rich CB and confirmed the importance of surface precipitation.

Molecular identification and phylogenetic analysis of Cryptosporidium, Hepatozoon and Spirometra in snakes from central China.

Snakes are popular as food and traditional medicine in China. However, information about parasitic and bacterial infections in snakes from China is scarce. We investigated the prevalence of selected zoonotic agents including Cryptosporidium, Hepatozoon and Spirometra, in snakes in central China from June to October in 2018 by PCR amplification using parasite-specific primers. PCR amplification and DNA sequencing showed that 10.1% (15/149) of snakes were positive for Cryptosporidium spp., while 2.7% (4/149) were positive for Hepatozoon. Additionally, we found 36.9% (55/149) of snakes were infected with Spirometra erinaceieuropaei. The spargana burden per infected snake ranged from 1 to 26. BLAST and phylogenetic analysis of small subunit ribosomal RNA (SSU rRNA) gene and 60-kDa glycoprotein (gp60) gene showed that the parasites belonged to Cryptosporidium parvum genotype IIdA15G1, C. baileyi, C. serpentis and a Hepatozoon species. We conclude that intensively farmed snakes excrete C. parvum and C. baileyi oocysts due to ingestion of infected feeder animals, and that wild snakes in central China were commonly infected with S. erinaceieuropaei, suggesting that eating improperly cooked snakes could be risky to human health.

High prevalence and genetic diversity of hepatitis B viruses in insectivorous bats from China.

Bats have been identified as the hosts of hepatitis B virus (HBV) in recent years and bats HBV can infect human hepatocyte. We investigated the prevalence and genetic diversity of HBV in bats in China. In this study, a total of 197 insectivorous bats belonging to 10 bat species were captured from karst caves in Mengyin County, Shandong Province and Xianning City, Hubei Province, China. PCR amplification indicated that in total 6.6% (13/197) bats were positive to HBVs. The HBV positive rate in bats was 7.1% (9/127) and 5.7% (4/70) in Shandong Province and Hubei Province, respectively. Phylogenetic analysis indicated that HBV from the two places were in the same cluster with 90.5%-99.5% homology, but distinct from bat HBVs from other places in China and other countries. We concluded that HBV was prevalent and genetic diversified in bats, supporting the hypothesis that bats may be the origin of primate hepadnaviruses.

Leptospira in Bats from Hubei Province, China, 2018.

To understand the potential role of bats in the epidemiology of Leptospira, we investigated the frequency of Leptospira in bats from Central China in 2018. Sixty bats belonging to four species of two families were captured from rural areas of Xianning City, Hubei Province, China. We extracted DNA from the kidneys of bats, and Leptospira spp. were determined by PCR amplification of the rrs2, flaB, lipL32, and secY genes. We found that 57% (34/60) of bats were positive for Leptospira with at least one of the four genes, and the positive rate of Leptospira in bats was 45% (27/60) with rrs2, 50% (30/60) with flaB, 15% (9/60) with lipL32, and 27% (16/60) with secY. Phylogenetic analysis indicated that bats were infected with two species of Leptospira, including Leptospira interrogans and Leptospira borgpetersenii. A high prevalence of Leptospira spp. in bats suggested that bats were important carriers of Leptospira in China.

Performance and mechanisms of emerging animal-derived biochars for immobilization of heavy metals.

Animal-derived biochars (ADBs) are a series of newly emerging biochars from the pyrolysis of animal carcasses. Their elemental composition and structural properties would be quite different to those of plant-based biochars and/or manure-derived biochars (MDBs). In this work, ADBs derived from cattle and swine carcasses were applied to investigate their sorption potentials and mechanisms for immobilization of Pb(II), Cd(II), Cu(II), and Hg(II). ADBs showed high calcium (Ca; >27%) and phosphorus (P; >15%) contents and relatively low carbon (C; <8%) content. Ca and P were present in the skeleton as calcium-phosphate complexes, dominantly as hydroxyapatite (HAP), which was identified by CP-MAS 31P NMR analysis. By contrast, the reference biochars derived from cattle, swine, and poultry manures possessed low Ca (<13%) and P (<4%) contents and relatively high C (>30%) content. Sorption experiments showed that ADBs were more effective in immobilizing Pb(II), Cd(II), and Cu(II) than MDBs. However, the affinities of ADBs for Hg(II) were lower than those of MDBs. Characterization analyses indicated that Pb(II) was effectively immobilized via precipitation, whereas ion exchange was the dominant mechanism in combining Cd(II) or Cu(II) with ADBs. ADBs are unsuitable for immobilizing Hg(II) because of the lack of oxygen-containing functional groups on the C surface, which can induce the reduction of Hg(II) to form Hg2Cl2. The immobilized heavy metals were difficult to be rereleased from ADBs in simulated acid rain environment. Hence, ADBs are promising biochars for immobilizing Pb(II), Cd(II), and Cu(II).

Clinic significance of markedly decreased α-klothoin women with preeclampsia.

Preeclampsia (PE) is a leading cause of maternal and perinatal morbidity and mortality. Klotho is a novel gene and the secret form, α-klotho (α-KL), is related to preeclampsia. We conducted this cross-sectional study in Wuhan, China. We used immunohistochemistry, real-time PCR, western blot, ELISA to measure α-KL expression in placenta and its secretion in maternal and umbilical cord serum, and analyzed correlations between α-KL level and other parameters in normal and preeclampsia pregnancy. We found that both mRNA and protein expression of placental α-KL in women with PE was significantly lower than that in normal pregnancy. Also, expression level of α-KL in both maternal and umbilical cord was markedly decreased in PE patients. Further analyses showed that serum α-KL exhibited positive association with fetal birth weight, and reverse association with oxidative stress and renal function markers. Receiver operating characteristic analysis suggested α-KL might be a potential predictor for preeclampsia.

α-Naphthoflavone modulates inflammatory response in adipocytes-macrophages interaction through NFκB signaling.

OBJECTIVE: Our previous study demonstrated that α-naphthoflavone (α-NF) inhibits mouse 3T3-L1 pre-adipocytes differentiation via PPARγ, a key transcription factor in adipogenesis. Due to the critical role of inflammation in adipogenesis, we speculated that the suppression role of α-NF in adipogenesis might involve in modulation of cytokines secretion raised by adipocyte differentiation cocktail. Therefore, the present study aims to investigate the role of α-NF in modulating of inflammatory response during adipocytes differentiation and adipocyte-macrophage interaction. METHODS: Conditioned medium from different doses of α-NF treated 10-day differentiated 3T3-L1 adipocytes were collected to culture RAW264.7 macrophages. Conditioned medium from activated macrophages and α-NF pre-treated macrophage were used to investigate the effects of α-NF in adipocytes differentiation. Cultured cells and medium were harvested for RT-PCR, Western blot and ELISA. RESULTS: α-NF dose-dependently decreased TNF-α and IL-6 and increased IL-10 expression induced by IDM (Insulin, dexamethasone, isobutylmethylxanthine) in 3T3-L1 pre-adipocytes. Conditioned medium from α-NF treated 3T3-L1 differentiated cells inhibited inflammatory response in mouse macrophage cell line RAW264.7 in contrast to IDM control medium. NFĸB activation elicited by IDM was suppressed by α-NF in a dose-response manner. Consequently, decreased TNF-α and increased IL-10 secretion, downstream targets of NFĸB signaling pathway, were observed with α-NF in macrophages. Finally, Conditioned medium from α-NF pre-treated, LPS-activated macrophages ameliorated the suppression of 3T3-L1 adipogenesis by LPS activated macrophages. CONCLUSION: Our results suggest that α-NF regulates inflammation response in both adipocytes and macrophages and adipocyte-macrophage interaction which contributes to pre-adipocyte differentiation.