Production and evaluation of anti-BP26 monoclonal antibodies for the serological detection of animal brucellosis.

Brucella BP26 proves to be a highly immunogenic antigen with excellent specificity in brucellosis detection. In China, the authorized use of the Bp26-deleted vaccine M5ΔBP26 for preventing small ruminant brucellosis highlights the importance of developing accurate detection methods targeting BP26, particularly for the diagnosis of differentiation between infected and vaccinated animals (DIVA). Using the traditional mouse hybridoma technique, we successfully obtained 12 monoclonal antibodies (mAbs) targeting BP26. The efficacy of these mAbs in detecting various animal brucellosis cases using the competitive ELISA method was evaluated. Among them, only the E10 mAb exhibited significant efficiency, being inhibited by 100, 97.62, and 100% of brucellosis-positive sera from cattle, small ruminants, and canines, respectively. The E10-based competitive enzyme-linked immunosorbent assay (cELISA) outperformed the BP26-based indirect enzyme-linked immunosorbent assay (iELISA) in accuracy, particularly for cattle and small ruminant brucellosis, with cELISA sensitivity reaching 97.62% compared to 64.29% for iELISA for small ruminants. Although cELISA showed slightly lower specificity than iELISA, it still maintained high accuracy in canine brucellosis detection. The epitope of mAb E10 was identified in the amino acid sequence QPIYVYPDDKNNLKEPTITGY, suggesting its potential as a diagnostic antigen for brucellosis. In conclusion, the E10-based cELISA presents an effective means of detecting animal brucellosis, particularly significant for DIVA diagnosis in China, where the BP26-mutant vaccine is widely used.

Development and evaluation of a triplex droplet digital PCR method for differentiation of M. tuberculosis, M. bovis and BCG.

Introduction: Tuberculosis, caused by Mycobacterium tuberculosis complex (MTBC), remains a global health concern in both human and animals. However, the absence of rapid, accurate, and highly sensitive detection methods to differentiate the major pathogens of MTBC, including M. tuberculosis, M. bovis, and BCG, poses a potential challenge. Methods: In this study, we have established a triplex droplet digital polymerase chain reaction (ddPCR) method employing three types of probe fluorophores, with targets M. tuberculosis (targeting CFP-10-ESAT-6 gene of RD1 and Rv0222 genes of RD4), M. bovis (targeting CFP-10-ESATs-6 gene of RD1), and BCG (targeting Rv3871 and Rv3879c genes of ΔRD1), respectively. Results: Based on optimization of annealing temperature, sensitivity and repeatability, this method demonstrates a lower limit of detection (LOD) as 3.08 copies/reaction for M. tuberculosis, 4.47 copies/reaction for M. bovis and 3.59 copies/reaction for BCG, without cross-reaction to Mannheimia haemolytica, Mycoplasma bovis, Haemophilus parasuis, Escherichia coli, Pasteurella multocida, Ochrobactrum anthropi, Salmonella choleraesuis, Brucella melitensis, and Staphylococcus aureus, and showed repeatability with coefficients of variation (CV) lower than 10%. The method exhibits strong milk sample tolerance, the LOD of detecting in spike milk was 5 × 103 CFU/mL, which sensitivity is ten times higher than the triplex qPCR. 60 clinical DNA samples, including 20 milk, 20 tissue and 20 swab samples, were kept in China Animal Health and Epidemiology Center were tested by the triplex ddPCR and triplex qPCR. The triplex ddPCR presented a higher sensitivity (11.67%, 7/60) than that of the triplex qPCR method (8.33%, 5/60). The positive rates of M. tuberculosis, M. bovis, and BCG were 1.67, 10, and 0% by triplex ddPCR, and 1.67, 6.67, and 0% by triplex qPCR, with coincidence rates of 100, 96.7, and 100%, respectively. Discussion: Our data demonstrate that the established triplex ddPCR method is a sensitive, specific and rapid method for differentiation and identification of M. tuberculosis, M. bovis, and BCG.

Rhizophagus irregularis and biochar can synergistically improve the physiological characteristics of saline-alkali resistance of switchgrass.

Inoculation of arbuscular mycorrhizal fungi (AMF) or biochar (BC) application can improve photosynthesis and promote plant growth under saline-alkali stress. However, little is known about the effects of the two combined on growth and physiological characteristics of switchgrass under saline-alkali stress. This study examined the effects of four treatments: (1) no AMF inoculation and no biochar addition (control), (2) biochar (BC) alone, (3) AMF (Rhizophagus irregularis, Ri) alone, and (4) the combination of both (BC+Ri) on the plant biomass, antioxidant enzymes, chlorophyll, and photosynthetic parameters of switchgrass under saline-alkali stress. The results showed that the above-ground, belowground and total biomass of switchgrass in the BC+Ri treatment group was significantly higher (+136.7%, 120.2% and 132.4%, respectively) than in other treatments compared with Control. BC+Ri treatment significantly increased plant leaves' relative chlorophyll content, antioxidant enzyme activity, and photosynthesis parameters. It is worth noting that the transpiration rate, stomatal conductance, net photosynthetic rate, PSII efficiency and other photosynthetic-related indexes of the BC+Ri treatment group were the highest (38% to 54% higher than other treatments). The fitting results of light response and CO2 response curves showed that the light saturation point, light compensation point, maximum carboxylation rate and maximum electron transfer rate of switchgrass in the Ri+BC treatment group were the highest. In conclusion, biochar combined with Ri has potential beneficial effects on promoting switchgrass growth under saline-alkali stress and improving the activity of antioxidant enzymes and photosynthetic characteristics of plants.

Insula uses overlapping codes for emotion in self and others.

In daily life, we must recognize others' emotions so we can respond appropriately. This ability may rely, at least in part, on neural responses similar to those associated with our own emotions. We hypothesized that the insula, a cortical region near the junction of the temporal, parietal, and frontal lobes, may play a key role in this process. We recorded local field potential (LFP) activity in human neurosurgical patients performing two tasks, one focused on identifying their own emotional response and one on identifying facial emotional responses in others. We found matching patterns of gamma- and high-gamma band activity for the two tasks in the insula. Three other regions (MTL, ACC, and OFC) clearly encoded both self- and other-emotions, but used orthogonal activity patterns to do so. These results support the hypothesis that the insula plays a particularly important role in mediating between experienced vs. observed emotions.

Coinoculation of arbuscular mycorrhizal fungi and rhizobia stimulates atrazine dissipation by changing the atrazine-degrading bacterial community at the soil aggregate scale.

As a potential low-cost and environmentally friendly strategy, bioremediation of herbicide polluted soil has attracted increasing attention. However, there is a lack of knowledge regarding the response of the atrazine-degrading bacterial community to coinoculation of arbuscular mycorrhizal (AM) fungi and rhizobia for atrazine dissipation. In this study, a pot experiment was conducted with AM fungi Glomus mosseae (AM), rhizobia Rhizobium trifolii TA-1 (R) and their coinoculation (AMR) with atrazine. In each treatment, the atrazine-degrading bacterial community of four soil size aggregates, namely large macroaggregates (LMa), small macroaggregates (SMa), microaggregates (Mia) and primary particles (P) were investigated. The results showed that the atrazine residue concentration was lowest in AMR, and that in LMa was also significantly lower than that in the other smaller aggregate sizes. Overall, inoculation, the aggregate fraction and their interaction had significant effects on soil TN, SOC, AP and pH. For the atrazine-degrading bacterial community, the Chao1 index increased with decreasing particle size, but the Shannon index decreased. Moreover, the abundances of the dominant atrazine-degrading bacterial genera Arthrobacter, Bacillus, Marmoricola and Nocardioides in the Mia and P particle size groups were greater than those in the LMa and SMa groups in each treatment. The bacterial communities in the Mia and P particle sizes in each treatment group were more complex. Therefore, coinoculation of AM fungi and rhizobia stimulated atrazine dissipation by changing the atrazine-degrading bacterial community, and the response of the atrazine-degrading bacterial community to each aggregate size varied depending on its distinct soil physicochemical properties.

Rhythmic attentional sampling in autism.

Individuals diagnosed with autism often display alterations in visual spatial attention toward visual stimuli, but the underlying cause of these differences remains unclear. Recent evidence has demonstrated that covert spatial attention, rather than remaining constant at a cued location, samples stimuli rhythmically at a frequency of 4-8 Hz (theta). Here we tested whether rhythmic sampling of attention is altered in autism. Participants were asked to monitor three locations to detect a brief target presented 300-1200 ms after a spatial cue. Visual attention was oriented to the cue and modified visual processing at the cued location, consistent with previous studies. We measured detection performance at different cue-target intervals when the target occurred at the cued location. Significant oscillations in detection performance were identified using both a traditional time-shuffled approach and a new autoregressive surrogate method developed by Brookshire in 2022. We found that attention enhances behavioral performance rhythmically at the same frequency in both autism and control group at the cued location. However, rhythmic temporal structure was not observed in a subgroup of autistic individuals with co-occurring attention-deficit/hyperactivity disorder (ADHD). Our results imply that intrinsic brain rhythms which organize neural activity into alternating attentional states is functional in autistic individuals, but may be altered in autistic participants who have a concurrent ADHD diagnosis.

Brain mechanisms underlying the emotion processing bias in treatment-resistant depression.

Depression is associated with a cognitive bias towards negative information and away from positive information. This biased emotion processing may underlie core depression symptoms, including persistent feelings of sadness or low mood and a reduced capacity to experience pleasure. The neural mechanisms responsible for this biased emotion processing remain unknown. Here, we had a unique opportunity to record stereotactic electroencephalography (sEEG) signals in the amygdala and prefrontal cortex (PFC) from 5 treatment-resistant depression (TRD) patients and 12 epilepsy patients (as control) while they participated in an affective bias task in which happy and sad faces were rated. First, compared with the control group, patients with TRD showed increased amygdala responses to sad faces in the early stage (around 300 ms) and decreased amygdala responses to happy faces in the late stage (around 600 ms) following the onset of faces. Further, during the late stage of happy face processing, alpha-band activity in PFC as well as alpha-phase locking between the amygdala and PFC were significantly greater in TRD patients compared to the controls. Second, after deep brain stimulation (DBS) delivered to bilateral subcallosal cingulate (SCC) and ventral capsule/ventral striatum (VC/VS), atypical amygdala and PFC processing of happy faces in TRD patients remitted toward the normative pattern. The increased amygdala activation during the early stage of sad face processing suggests an overactive bottom-up processing system in TRD. Meanwhile, the reduced amygdala response during the late stage of happy face processing could be attributed to inhibition by PFC through alpha-band oscillation, which can be released by DBS in SCC and VC/VS.

Performance of atrazine adsorption behavior and microbial community structure in Mollisol aggregate fraction.

Owing to complex pore systems and chemical substances, soil aggregates provide a spatially heterogeneous microenvironment for adsorption capacity and microbial survival. As the widely used pesticide in farmlands, atrazine environmental behavior is not well known at the aggregate scale. In this study, Mollisol soil samples were sieved into four aggregate-size classes: large macroaggregates (>2 mm, LMa), small macroaggregates (1-2 mm, SMa), microaggregates (0.25-1 mm, Mia) and primary particles (<0.25 mm, P). The pore characteristics of each aggregate fraction was visualized by non-invasive X-ray three-dimensional microscopic computed tomography (3D-CT) combined with pore network extraction. The adsorption kinetics of atrazine in each aggregate-size fraction can be described well by a pseudo-second-order kinetic model. The adsorption isothermal process of atrazine can be better fitted by the Langmuir isotherm model than Freundlich isotherm model. There was an obvious linear correlation between the maximum atrazine adsorption capacity and aggregate SOC content as well as TN. In addition, the abundance of bacteria, actinomycetes and anaerobic bacteria in P was totally higher than those in SMa and Mia. Although pH is strongly linked to the bacterial community in the aggregate fraction, aggregate particle size explained 18 % for shaping the microbial community. Therefore, chemical properties and pore characteristics of each soil aggregate fraction both contributed to performance of atrazine adsorption behavior and microbial community.

Cocultivation with Solanum nigrum and inoculation with Rhizophagus intraradices can improve plant photosynthesis and antioxidant defense to alleviate cadmium toxicity to soybean.

High Cd pollution can damage plant physiology and seriously threaten ecological security and human health. Therefore, we designed a cropping system, arbuscular mycorrhizal fungi (AMF) - soybean - Solanum nigrum L., to solve the high Cd pollution problem in an environmentally and economically friendly way. The results showed that AMF were able to break free from the constraints of cocultivation and still promote plant photosynthesis and growth in combined treatments to resist Cd stress. In addition, cocultivation combined with AMF improved the antioxidant defense to scavenge reactive oxygen species by promoting the production of antioxidant enzymes and nonenzyme substances in host plants. The glutathione content in soybean and the catalase activity in nightshade were recorded at the highest values under cocultivation combined with AMF treatment, which were 23.68% and 129.12% higher than those of monoculture without AMF treatments. The improvement in antioxidant defense alleviated oxidative stress, which was manifested by the reduction in Cd dense electronic particles in the ultrastructure and a 26.38% decrease in MDA content. Furthermore, this cropping mode combined the advantages of cocultivation to improve the Cd extraction efficiency and Rhizophagus intraradices to limit Cd accumulation and transport so that Cd was more accumulated and restricted in the roots of the cocultivated Solanum nigrum L., and the Cd concentration in soybean beans was reduced by 56% compared with the soybean monoculture without AMF treatment. Therefore, we suggest that this cropping system is a comprehensive and mild remediation technology suitable for highly Cd-contaminated soil.

Arbuscular mycorrhizal fungi influence the uptake of cadmium in industrial hemp (Cannabis sativa L.).

Phytoremediation is currently a more environmentally friendly and economical measure for the remediation of cadmium (Cd) contaminated soil. Heavy metal-resistant plant species, Cannabis sativa L. was inoculated with Rhizophagus irregularis to investigate the mechanisms of mycorrhizal in improving the Cd remediation ability of C. sativa. The results showed that after inoculation with R. irregularis, C. sativa root Cd contents increased significantly, and leaf Cd enrichment decreased significantly. At the transcriptional level, R. irregularis down-regulated the expression of the ABC transporter family but up-regulated differentially expressed genes regulating low molecular weight organic acids. The levels of malic acid, citric acid, and lactic acid were significantly increased in the rhizosphere soil, and they were significantly and strongly related to oxidizable Cd concentrations. Then citric acid levels were considerably and positively connected to exchangeable Cd concentrations. Our findings revealed that through regulating the movement of root molecules, arbuscular mycorrhizal fungus enhanced the heavy metal tolerance of C. sativa even more, meanwhile, they changed the Cd chemical forms by altering the composition of low molecular weight organic acids, which in turn affected soil Cd bioavailability.

Top-down modulation and cortical-AMG/HPC interaction in familiar face processing.

Humans can accurately recognize familiar faces in only a few hundred milliseconds, but the underlying neural mechanism remains unclear. Here, we recorded intracranial electrophysiological signals from ventral temporal cortex (VTC), superior/middle temporal cortex (STC/MTC), medial parietal cortex (MPC), and amygdala/hippocampus (AMG/HPC) in 20 epilepsy patients while they viewed faces of famous people and strangers as well as common objects. In posterior VTC and MPC, familiarity-sensitive responses emerged significantly later than initial face-selective responses, suggesting that familiarity enhances face representations after they are first being extracted. Moreover, viewing famous faces increased the coupling between cortical areas and AMG/HPC in multiple frequency bands. These findings advance our understanding of the neural basis of familiar face perception by identifying the top-down modulation in local face-selective response and interactions between cortical face areas and AMG/HPC.

Comparison of BP26, Omp25 and Omp31 and a Multiepitope-Based Fusion Protein in the Serological Detection of Canine Brucellosis.

Background: Brucellosis is one of the most important zoonotic diseases in the world. Canine brucellosis, caused mainly by Brucella canis, is seriously neglected, and there is a lack of accurate diagnostic tools. Methods: In this study, to compare BP26, Omp25, Omp31 and a multiepitope-based fusion protein in the serological detection of canine brucellosis, using 34 brucellosis-positive dog sera and 62 negative control sera, the Brucella outer membrane proteins Omp31, BP26, Omp25 and a multiepitope-based fusion protein were evaluated by iELISA for their potential use as antigens in the serological diagnosis of canine brucellosis. Results: The results showed that the multiepitope-based fusion protein performed best in distinguishing brucellosis-positive and brucellosis-negative dog sera, with a positive predictive value (PPV) of 100% and a negative predictive value (NPV) of 98.41%. BP26 and Omp31 showed excellent sensitivity in detecting brucellosis-positive dog sera, but their cross reaction to sera infected with Vibrio parahaemolyticus and Listeria monocytogenes may hinder their application as diagnostic reagents. Omp25 lacked sufficient sensitivity and showed limited ability in distinguishing positive and negative dog sera. Conclusion: The multiepitope-based fusion protein can be used as an ideal antigen for serologically diagnosing canine brucellosis currently prevalent worldwide.

The impact of Mycobacterium tuberculosis complex in the environment on one health approach.

Tuberculosis caused by the Mycobacterium tuberculosis complex (MTBC) has become one of the leading causes of death in humans and animals. Current research suggests that the transmission of MTBC in the environment indirectly transmit to humans and animals with subsequent impact on their wellbeing. Therefore, it is of great significance to take One Health approach for understanding the role of MTBC in not only the interfaces of humans and animals, but also environment, including soil, water, pasture, air, and dust, etc., in response to the MTBC infection. In this review, we present the evidence of MTBC transmission from environment, as well as detection and control strategies in this interface, seeking to provide academic leads for the global goal of End Tuberculosis Strategy under multidisciplinary and multisectoral collaborations.

Honokiol improves depression-like behaviors in rats by HIF-1α- VEGF signaling pathway activation.

Increasing evidence indicates that the pathogenesis of depression is closely linked to impairments in neuronal synaptic plasticity. Honokiol, a biologically active substance extracted from Magnolia Officinalis, has been proven to exert significant antidepressant effects. However, the specific mechanism of action remains unclear. In this study, PC12 cells and chronic unpredictable mild stress (CUMS) model rats were used to explore the antidepressant effects and potential mechanisms of honokiol in vitro and in rats. In vitro experiment, a cell viability detection kit was used to screen the concentration and time of honokiol administration. PC12 cells were administered with hypoxia-inducible factor-1α (HIF-1α) blocker, 2-methoxyestradiol (2-ME), and vascular endothelial growth factor receptor 2 (VEGFR-2) blocker, SU5416, to detect the expression of HIF-1α, VEGF, synaptic protein 1 (SYN 1), and postsynaptic density protein 95 (PSD 95) by western blotting. In effect, we investigated whether the synaptic plasticity action of honokiol was dependent on the HIF-1α-VEGF pathway. In vivo, behavioral tests were used to evaluate the reproducibility of the CUMS depression model and depression-like behaviors. Molecular biology techniques were used to examine mRNA and protein expression of the HIF-1α-VEGF signaling pathway and synaptic plasticity-related regulators. Additionally, molecular docking techniques were used to study the interaction between honokiol and target proteins, and predict their binding patterns and affinities. Experimental results showed that honokiol significantly reversed CUMS-induced depression-like behaviors. Mechanically, honokiol exerted a significant antidepressant effect by enhancing synaptic plasticity. At the molecular level, honokiol can activate the HIF-1α-VEGF signaling pathway in vitro and in vivo, as well as promote the protein expression levels of SYN 1 and PSD 95. Taken together, the results do not only provide an experimental basis for honokiol in the clinical treatment of depression but also suggest that the HIF-1α-VEGF pathway may be a potential target for the treatment of depression.

Antidepressant-Like Effect and Mechanism of Ginsenoside Rd on Rodent Models of Depression.

Background: There is growing evidence to suggest that ginsenoside Rd (GRd) has a therapeutic effect on depression, but the specific mechanisms behind its activity require further study. Objective: This study is designed to investigate the antidepressant-like effect and underlying mechanisms of GRd. Methods: In this study, the behavioral despair mouse model of depression and chronic unpredictable mild stress (CUMS) rat model of depression were established to explore the effects of GRd on depression-like behavior and its underlying mechanisms. Behavioral tests were used to evaluate the replication of animal models and depression-like behaviors. The hypoxia-inducible factor-1α (HIF-1α) blocker 2-methoxyestradiol (2-ME) was injected to determine the role of HIF-1α in the antidepressant-like effect of GRd. In addition, molecular biology techniques were used to determine the mRNA and protein expression of HIF-1ɑ signaling pathway and synaptic plasticity-related regulators, that is synapsin 1 (SYN 1) and postsynaptic density protein 95 (PSD 95). In silico binding interaction studies of GRd with focused target proteins were performed using molecular docking to predict the affinity and optimal binding mode between ligands and receptors. Results: Our data show that GRd significantly reversed depression-like behavior and promoted mRNA and protein expression of HIF-1ɑ signaling pathway and synaptic plasticity-related regulators. However, the antidepressant-like effect of GRd disappeared upon inhibition of HIF-1α expression following administration of 2-ME. Furthermore, molecular docking results showed that GRd possessed significant binding affinity for HIF-1α, VEGF, and VEGFR-2. Conclusion: Our results show that GRd exhibits significant antidepressant-like effect and that HIF-1α signaling pathway is a promising target for the treatment of depression.

Cross-sectional study on risk factors associated with porcine epidemic diarrhea virus infection in pig farms in Junan county, China.

Porcine epidemic diarrhea (PED) is a highly contagious, intestinal infectious disease in pigs, characterized by severe diarrhea, vomiting and dehydration. PED is widely epidemic in China as well as in many eastern Asian and America countries, causing tremendous losses in pig industry. However, little was known about the disease frequency and the associated risk factors of PED in pig farms. A cross-sectional study was conducted to estimate the herd prevalence and to identify the potential risk factors of porcine epidemic diarrhea virus (PEDV) infection of pig farms (≥100 pigs) in Junan county, China. A two-stage random sampling strategy was adopted, and a total of 751 fecal samples from suckling piglets of 82 farms were collected and tested by RT-PCR and sequencing for PEDV. Meanwhile, information on the putative risk factors of PEDV infection of those farms were collected in forms of questionnaires, followed by a descriptive analysis, univariable and multivariable logistic regression analysis. The results showed that the herd-level true prevalence of PEDV infection of pig farms in Junan was 35.16 % (95 %CI: 22.91-53.89); there were two variables significantly associated with PEDV infection, which were 'having more than 1000 slaughter pigs per annum' (OR = 5.42, 95 %CI: 1.19-24.72), and 'Weaning at 21-25 days' old' (OR = 4.50, 95 %CI:1.25-16.20). The research suggested that PED was highly endemic in pig farms in Junan county; larger herd size and weaning at an earlier age were potential risk factors associated with PEDV infection in pig farms in Junan. This study set an example in the research on herd-level prevalence of PEDV infection and risk factors associated with PEDV infection, and the results were of practical significance for the future planning of prevention and control of PED in Junan or other areas of China.

Synthesis of hierarchical and flower-like TiO2 nanowire microspheres as biocompatible cell carriers.

Fibrous materials are of great interest in the development of tissue regenerative matrix. However, synthesis of inorganic fibrous microspheres as cell carriers is a great challenge. In this study, we for the first time report on the synthesis of novel hierarchical and flower-like TiO2 nanowire (NW) microspheres as biocompatible cell carriers. TiO2 NW microspheres were synthesized through in situ alkali hydrothermal treatment of the TiO2 nanoparticle (NP) microspheres and their microstructure, formation mechanism and in vitro biocompatibility were evaluated. SEM observations show that the resulting TiO2 NW microspheres were constructed by a large number of NWs with the diameter of 10-20 nm and exhibited a flower-like and hierarchical morphology with the diameter of 400-600 µm. XRD patterns indicate that TiO2 NW microspheres were constructed by both rutile and anatase phase of TiO2. FT-IR spectra reveal that Ti-O-Ti bonds were involved in TiO2 NW microspheres. In vitro biocompatibility was evaluated by seeding the fibroblast L929 cells on the microspheres. A conventional MTT assay quantitatively indicates that the TiO2 NW microspheres favored adhesion and proliferation of cells and were biocompatible, while SEM observations qualitatively confirmed that the cells were well grown on the surface of TiO2 NW microspheres. Thus, the as-synthesized TiO2 NW microspheres would be applicable to novel and biocompatible cell carriers.

High prevalence of extrapulmonary tuberculosis in dairy farms: Evidence for possible gastrointestinal transmission.

Bovine tuberculosis (bTB) caused by Mycobacterium bovis (M. bovis) represents one of major zoonotic diseases among cattle, it also affects the health of human, other domestic animals and wild life populations. Inhalation of infected aerosol droplets is considered as the most frequent route of the infection. This study aims to investigate the current forms of tuberculosis in cattle and identify the possible transmission modes in dairy farms of China. 13,345 cows from eight dairy farms in three provinces were comprehensively diagnosed by a multitude of assays, including SIT, CIT, IFN-γ assay and ELISA. It has been indicated that advanced infection of bTB was found in 752 (5.64%) cattle, suggesting a high prevalence of tuberculosis in these dairy farms. In the necropsy examination of 151 positive cattle, typical bTB lesions were observed in 131 cattle (86.75%), of which, notably, 90.84% lesions appeared in liver, spleen, mesenteric lymph nodes, mammary lymph nodes and other organs, taking up a large proportion among cattle with advanced bTB infection. 71.26% extrapulmonary tuberculosis (EPTB) was related to gastrointestinal system. M. bovis nucleic acid was further found in milk and feces samples and M. bovis was even isolated from milk samples. Phylogenetic analysis based on whole genome sequencing unraveled that six isolates were closely related to M. bovis AF2122/97 originated from UK, whereas four isolates shared close relation to M. bovis 30 from China, respectively. Our data demonstrate that the increase of EPTB transmitted by digestive tract is implicated in the current high prevalence rate of bTB in China, which also provides leads for bTB control in other countries with high prevalence of bTB in the future.

Clinical Validation of Two Recombinase-Based Isothermal Amplification Assays (RPA/RAA) for the Rapid Detection of African Swine Fever Virus.

African swine fever (ASF), caused by African swine fever virus (ASFV), is a devastating infectious disease of domestic pigs and wild boars, and has tremendous negative socioeconomic impact on the swine industry and food security worldwide. It is characterized as a notifiable disease by World Organisation for Animal Health (OIE). No effective vaccine or treatment against ASF has so far been available. Early detection and rapid diagnosis are of potential significance to control the spread of ASF. Recombinase-based isothermal amplification assay, recombinase polymerase amplification (RPA) developed by TwistDx (Cambridge, United Kingdom) or recombinase-aided amplification (RAA) by Qitian (Wuxi, China), is becoming a molecular tool for the rapid, specific, and cost-effective identification of multiple pathogens. In this study, we aim to investigate if RPA/RAA can be a potential candidate for on-site, rapid and primary detection of ASFV. A panel of 152 clinical samples previously well-characterized by OIE-recommended qPCR was enrolled in this study, including 20 weak positive (Ct value ≥ 30) samples. This panel was consisted of different types, such as EDTA-blood, spleen, lung, lymph node, kidney, tonsil, liver, brain. We evaluated two recombinase-based isothermal amplification assays, RPA or RAA, by targeting the ASFV B646L gene (p72), and validated the clinical performance in comparison with OIE real-time PCR. Our result showed that the analytical sensitivity of RPA and RAA was as 93.4 and 53.6 copies per reaction, respectively at 95% probability in 16 min, at 39°C. They were universally specific for all 24 genotypes of ASFV and no cross reaction to other pathogens including Classical swine fever virus (CSV), Foot-and-mouth disease virus (FMDV), Pseudorabies virus, Porcine circovirus 2 (PCV2), Porcine Reproductive and respiratory syndrome virus (PPRSV). The results on detection of various kinds of clinical samples indicated an excellent diagnostic agreement between RPA, RAA and OIE real-time PCR method, with the kappa value of 0.960 and 0.973, respectively. Compared to real-time PCR, the specificity of both RPA and RAA was 100% (94.40% ∼ 100%, 95% CI), while the sensitivity was 96.59% (90.36% ∼ 99.29%, 95% CI) and 97.73% (92.03% ∼ 99.72%, 95% CI), respectively. Our data demonstrate that the developed recombinase-based amplification assay (RPA/RAA), promisingly equipped with field-deployable instruments, offers a sensitive and specific platform for the rapid and reliable detection of ASFV, especially in the resource-limited settings for the purpose of screening and surveillance of ASF.

Facile synthesis, microstructure, formation mechanism, in vitro biocompatibility, and drug delivery property of novel dendritic TiO2 nanofibers with ultrahigh surface area.

In this study, novel dendritic TiO2 nanofibers (D-Ti NFs) with ultrahigh surface area were for the first time synthesized through in situ hydrothermal treatment of the electrospun TiO2 nanofibers (E-Ti NFs) and their microstructure, formation mechanism, in vitro biocompatibility and drug delivery property were investigated. SEM observations showed that D-Ti NFs had dendritic and fibrous morphology with the controllable diameter of 899 ± 26 nm to 1955 ± 64 nm. TEM observations showed the individual D-Ti NF was assembled by numerous primary nanowires with the diameter of 7 ± 1 nm. XRD pattern showed that the D-Ti NFs displayed the characteristic peaks at 24.1°, 28.1°, and 48.3°, all of which were assigned to hydrogen titanium oxide hydrate. BET measurement indicated that D-Ti NFs had the ultrahigh specific surface area of 213.41 m2/g. D-Ti NFs showed good in vitro biocompatibility when exposed to Hela cells. After soaked in solution of tetracycline hydrochloride (TH, one of the representative antibiotics, model drug), D-Ti NFs supported the loading of TH with the loading efficiency of 63.73 ± 4.61%. TH-loaded D-Ti NFs supported a sustained release model for TH when immersed in the phosphate buffer saline. After separately incubated with two representative types of bacteria: Escherichia coli and Staphylococcus aureus, TH-loaded D-Ti NFs showed excellent antibacterial property to inhibit the growth of the bacteria, indicating that the released TH was biologically active.