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Abstract Repair welding technology is widely used in the manufacturing and maintenance of rail transit equipment to repair welding defects. However, repair welding induces modifications in joint performance, and it is necessary to study the microstructure evolution behavior to reveal the reasons. In this study, the effects of multiple repair welding on the microstructure, mechanical, and fatigue properties of 06Cr19Ni10 stainless steel samples were studied. The surface texture and fracture morphology analyses of the samples were conducted by optical microscopy (OM), scanning electron microscopy (SEM), and its equipped backscattered electron diffraction (EBSD) technique. The mechanical and fatigue properties of the samples with different repair welding times were further obtained by hardness, tensile, and fatigue tests. The results show an increase in the grain size and the substructure content in the heat-affected zone (HAZ), and the austenite orientation is changed, attributable to multiple repair welding. Multiple heat inputs result in a significant increase in hardness from 165 HV to 185 HV, a noticeable decrease in tensile strength and elongation, and an upward trend in yield strength. Under the constant stress level, the heat input of multiple repair welding causes a decrease in the fatigue life and significantly reduces toughness in the instantaneous fracture zone of the secondary repair sample.

Abstract Micro-expressions are spontaneous, rapid and subtle facial movements that can hardly be suppressed or fabricated. Micro-expression recognition (MER) is one of the most challenging topics in affective computing. It aims to recognize subtle facial movements which are quite difficult for humans to perceive in a fleeting period. Recently, many deep learning-based MER methods have been developed. However, how to effectively capture subtle temporal variations for robust MER still perplexes us. We propose a counterfactual discriminative micro-expression recognition (CoDER) method to effectively learn the slight temporal variations for video-based MER. To explicitly capture the causality from temporal dynamics hidden in the micro-expression (ME) sequence, we propose ME counterfactual reasoning by comparing the effects of the facts w.r.t. original ME sequences and the counterfactuals w.r.t. counterfactually-revised ME sequences, and then perform causality-aware prediction to encourage the model to learn those latent ME temporal cues. Extensive experiments on four widely-used ME databases demonstrate the effectiveness of CoDER, which results in comparable and superior MER performance compared with that of the state-of-the-art methods. The visualization results show that CoDER successfully perceives the meaningful temporal variations in sequential faces.

To study the Mittag-Leffler projective synchronization (MLPS) problem of fractional-order fuzzy neural networks (FOFNNs), in this work we introduced the FOFNNs model. On this basis, we discussed the MLPS of uncertain fractional-order fuzzy complex valued neural networks (FOFCVNNs) with distributed and time-varying delays. Utilizing Banach contraction mapping principle, we proved the existence and uniqueness of the model solution. Moreover, employing the construction of a new hybrid controller, an adaptive hybrid controller, and the fractional-order Razumikhin theorem, algebraic criteria was obtained for implementing MLPS. The algebraic inequality criterion obtained in this article improves and extends the previously published results on MLPS, making it easy to prove and greatly reducing the computational complexity. Finally, different Caputo derivatives of different orders were given, and four numerical examples were provided to fully verify the accuracy of the modified criterion.

Abstract Background Chlorophenoxy compounds represent a group of selective herbicides widely used around the world. Chlorophenoxy herbicides are toxic, chemically stable, and can migrate into groundwater through soil leaching, posing a significant threat to drinking water safety and human health. Chlorophenoxy herbicides in groundwater aquifers are subject to anaerobic processes; however, the pathway and microbiology involved in the attenuation of chlorophenoxy herbicides under anaerobic condition are largely unknown. Here, the anaerobic degradation process of 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), a typical chlorophenoxy herbicide, was investigated. Results The initial 52.5 ± 2.3 μM 2,4,5-T was completely degraded by a sediment-derived microbial consortium, with 3,4-dichlorophenol, 2,5-dichlorophenol, 3-chlorophenol (3-CP) and phenol being identified as the intermediate products. Reductive dechlorination of 3-CP to phenol and the subsequent elimination of phenol were the key transformation steps in the overall degradation process of 2,4,5-T. Amplicon sequencing suggested that Dehalobacter, Sulfuricurvum, Bacteroides, Acetobacterium, and Clostridium sensu stricto 7 might contribute to the transformation of 2,4,5-T to phenol, and Smithella, Syntrophorhabdus, Methanofollis and Methanosaeta likely cooperated to accomplish the complete mineralization of phenol. Conclusions This study reported the anaerobic degradation of 2,4,5-T via reductive dechlorination and the subsequent syntrophic metabolization of phenol, an intermediate product transformed from 2,4,5-T. Dehalobacter was identified as the organohalide-respiring population catalyzing the reductive dechlorination reaction. Syntrophorhabdus and methanogenic populations were likely involved in anaerobic phenol oxidation and facilitated the complete mineralization of 2,4,5-T.

Abstract Light-emitting diodes (LEDs) based on perovskite semiconductor materials with tunable emission wavelength in visible light range as well as narrow linewidth are potential competitors among current light-emitting display technologies, but still suffer from severe instability driven by electric field. Here, we develop a stable, efficient and high-color purity hybrid LED with a tandem structure by combining the perovskite LED and the commercial organic LED technologies to accelerate the practical application of perovskites. Perovskite LED and organic LED with close photoluminescence peak are selected to maximize photon emission without photon reabsorption and to achieve the narrowed emission spectra. By designing an efficient interconnecting layer with p-type interface doping that provides good opto-electric coupling and reduces Joule heating, the resulting green emitting hybrid LED shows a narrow linewidth of around 30 nm, a peak luminance of over 176,000 cd m−2, a maximum external quantum efficiency of over 40%, and an operational half-lifetime of over 42,000 h.

Abstract Background Multiple species of the genera Cytauxzoon and Hepatozoon can infect wild felines, but the diversity of these and other apicomplexan parasites in Eurasian lynx is scarcely known. The aim of this study was to detect Cytauxzoon and Hepatozoon species with molecular methods in Eurasian lynxes and their ticks in northwestern China. Methods DNA was extracted from the heart, liver, spleen, lung, and kidney samples of three Eurasian lynxes as well as from their five ixodid ticks. These DNA samples were screened with polymerase chain reactions (PCRs) for Cytauxzoon with the partial cytochrome b gene (CytB), cytochrome c oxidase subunit I gene (COI), and small subunit ribosomal RNA gene (18S rRNA), and Hepatozoon with three different fragments of small subunit ribosomal RNA gene (18S rRNA). PCR products were sequenced, aligned, and phylogenetically analyzed. Results One adult female of Eurasian lynx (#1, adult female) was co-infected with Cytauxzoon manul and Hepatozoon felis genotype I, while an adult male lynx (#2) was infected with C. manul. Interestingly, H. felis genotype I was both detected in a male cub (#3) and two out of five infesting Hyalomma asiaticum ticks. Conclusions For the first time, Cytauxzoon manul is reported here from Eurasian lynx. In addition, H. felis has not been known to occur in this host species in China and Central Asia. Thus, the findings of this study extend our knowledge on the geographical distribution and host range of these haemoprotozoan parasites. Moreover, this is also the first evidence of C. manul and H. felis co-infection in Eurasian lynx. Graphical Abstract

Abstract Background Ticks are blood-feeding significant arthropods that can harbour various microorganisms, including pathogens that pose health risks to humans and animals. Tick-symbiont microorganisms are believed to influence tick development, but the intricate interactions between these microbes and the relationships between different tick-borne microorganisms remain largely unexplored. Results Based on 111 tick pool samples presenting questing and engorged statuses including 752 questing tick and 1083 engorged tick from cattle and goats, which were collected in two types of geographic landscape (semi-desert and alpine meadow). We observed significant variations in the composition of tick-borne microorganisms across different environments and blood-engorgement statuses, with a pronounced divergence in symbionts compared to environmental bacteria. Metabolic predictions revealed over 90 differential pathways for tick-borne microorganisms in distinct environments and more than 80 metabolic variations in response to varying blood engorgement statuses. Interestingly, nine pathways were identified, particularly related to chorismate synthesis and carbohydrate metabolism. Moreover, microbial network relationships within tick-borne microorganism groups were highly distinct across different environments and blood-engorgement statuses. The microbial network relationships of symbionts involve some pathogenic and environmental microorganisms. Regression modelling highlighted positive correlations between the Coxiella symbiont and related pathogens, while some environmental bacteria showed strong negative correlations with Coxiella abundance. We also identified commensal bacteria/pathogens in bacterial cooccurrence patterns. Furthermore, we tested pathogenic microorganisms of each tick sample analysis revealed that 86.36% (1601/1855) of the tick samples carried one or more pathogenic microorganisms, The total carrier rate of bacterial pathogens was 43.77% ((812/1855). Most blood samples carried at least one pathogenic microorganism. The pathogens carried by the ticks have both genus and species diversity, and Rickettsia species are the most abundant pathogens among all pathogens. Conclusion Our findings underscore that the bacterial pattern of ticks is dynamic and unstable, which is influenced by the environment factors and tick developmental characteristics.

ABSTRACT: This study aimed to investigate how various selenium sources affect the intestinal health of broiler chickens. A total of 384, one-day-old Arbor Acres broilers were weighed and randomly allocated to four treatment groups. The control diet was a basal diet added with: 0.2 mg/kg Sodium Selenite (SS-control), 0.2 mg/kg Selenium nano-particles (Nano-Se), 0.2 mg/kg Selenomethionine (SeMet), and 0.2 mg/kg Selenocysteine (Sec) as the treatments. The results indicated that Nano-Se and SeMet were effective in enhancing the villus height (VH) and the villus height/crypt depth ratio (VH/CD) in the jejunum compared to (SS) (P < 0.05). The inclusion of Nano-Se into the diets increased the mRNA levels of zonula occluden-1 (ZO-1), ZO-2, Occludin, Claudin-1, and Claudin-3 compared to the SS diet (P < 0.05). The SeMet increased the levels of ZO-1 and Claudin-3 compared to the SS (P < 0.05). Moreover, SeMet upregulated the marker genes of intestinal enteroendocrine cells, stem cells, and epithelial cells compared to the SS diet (P < 0.05). However, supplementation of Nano-Se reduced the mRNA levels of interleukin 1β (IL-1β), and IL-8 and the concentration of reactive oxygen species (ROS) in the jejunum compared to the SS (P < 0.05). The Nano-Se and SeMet also increased the protein levels of CAT and SOD compared to the SS and Sec diet (P < 0.05). The number of the goblet cells and Mucin-2 (Muc2) levels were the highest in the Nano-Se group (P < 0.05). The protein expression levels of goblet cell differentiation regulator (v-myc avian myelocytomatosis viral oncogene homolog, c-Myc) were highest in the Nano-Se compared to the SS diet (P < 0.05). The Nano-Se decreased the mRNA and protein levels of NLRP3 signaling pathway-related genes compared to the SS diet (P < 0.05). In conclusion, our study demonstrated that Nano-Se and SeMet are better at improving the intestinal health of 21-day-old broilers. Additionally, Nano-Se demonstrated superior antioxidant and anti-inflammatory effects, promoting the development of intestinal goblet cells by modifying the NLRP3 signaling pathway.

Fleas are known to be vectors for a variety of pathogens in veterinary medicine. However, no information is available on the presence of Bartonella and Trypanosomatidae in fleas of the long-tailed ground squirrel (LTGR, Spermophilus undulatus). The present study shows detection of these pathogens in LTGR fleas. During 2022–2023, a total of 396 fleas were collected from 91 LTGRs in 4 alpine regions of Xinjiang Uygur Autonomous Region (northwestern China) and grouped into 54 flea pools. Flea species were identified according to morphological characteristics and molecular data. In addition, all flea samples were analyzed for Bartonella with amplification and sequencing of a 380-bp part of the gltA gene and Trypanosomatidae with targeting the 18S rRNA (850-bp) and gGAPDH (820-bp) genes. The flea species included Frontopsylla elatoides elatoides (203), Neopsylla mana (49), and Citellophilus tesquorum dzetysuensis (144). Of 54 flea pools, seven (12.96%) tested positive for Bartonella, and three (5.56%) were positive for Trypanosomatidae. Based on BLASTn and phylogenetic analyses, i) Bartonella washoensis in F. elatoides elatoides and C. tesquorum dzetysuensis, and Bartonella rochalimae in F. elatoides elatoides were identified. Interestingly, a new haplotype within the species Ba. washoensis was discovered in C. tesquorum dzetysuensis; and ii) Blechomonas luni was confirmed in C. tesquorum dzetysuensis and Trypanosoma otospermophili in F. elatoides elatoides. Two Bartonella species and two Trypanosomatidae members were discovered for the first time in fleas from LTGRs. This study broadens our understanding of the geographic distribution and potential vectors for Bartonella and Trypanosomatidae.

Abstract Background In the family Trypanosomatidae, the genus Trypanosoma contains protozoan parasites that infect a diverse range of hosts, including humans, domestic animals, and wildlife. Wild rodents, as natural reservoir hosts of various pathogens, play an important role in the evolution and emergence of Trypanosomatidae. To date, no reports are available on the trypanosomatid infection of pikas (Lagomorpha: Ochotonidae). Methods In this study, Mongolian pikas and their fleas were sampled at the China–Mongolia border, northwestern China. The samples were analyzed with polymerase chain reaction (PCR) and sequencing for the presence of Trypanosomatidae on the basis of both the 18S ribosomal RNA (18S rRNA) gene and the glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) gene. The morphology of trypomastigotes was also observed in peripheral blood smears by microscopy. Results Molecular and phylogenetic analyses revealed a new genotype of the Trypanosoma lewisi clade that was found both in pika blood and flea samples. This genotype, which probably represents a new species, was provisionally designated as “Trypanosoma sp. pika”. In addition, a novel genotype belonging to the genus Blechomonas of Trypanosomatidae was detected in fleas. On the basis of its molecular and phylogenetic properties, this genotype was named Blechomonas luni-like, because it was shown to be the closest related to B. luni compared with other flea-associated trypanosomatids. Conclusions To the best of our knowledge, this is the first study to report any trypanosomatid species in Mongolian pikas and their fleas. Further studies are needed to investigate the epidemiology of these protozoan parasites, as well as to evaluate their pathogenicity for humans or domestic animals. Graphical Abstract

Abstract To investigate the efficacy and safety of drug-eluting bead-transarterial chemoembolization (DEB-TACE) combined with systemic chemotherapy in HR+/Her2− locally advanced breast cancer (LABC) patients. A controlled study was conducted on LABC patients treated at Jianyang People’s Hospital and the First Affiliated Hospital of Chengdu Medical College from December 2020 to June 2022. The patients were randomly divided into the experimental group and the control group. The experimental group received DEB-TACE combined with the TAC regimen (175 mg/m2 paclitaxel-loaded albumin, 50 mg/m2 Doxorubicin, and 500 mg/m2 cyclophosphamide), while the control group received the TAC regimen intravenously. The therapeutic efficacy was evaluated using the mRECIST criteria. Statistical analysis was performed using SPSS 22.0 software, and baseline characteristics, overall response rate (ORR), pathological complete response (PCR), adverse reactions, and complications were compared between the two groups using paired t-test and chi-square test. A total of 60 patients were included, with 30 patients in the experimental group (50%) and 30 patients in the control group (50%). After the first treatment, the ORR was 90% in the experimental group and 60% in the control group (P

Abstract Background Rodents play an important role in the life cycle of ixodid and argasid ticks, particularly as hosts of larvae and nymphs. The great gerbil (Rhombomys opimus), the preferred prey item of several carnivores (e.g. the red fox and marbled polecat), is the dominant rodent species in the Gurbantunggut Desert in northwestern China. The aim of this study was to investigate tick species associated with different hosts in the habitat of great gerbils, including wildlife and livestock. Methods During 2018–2023, ticks were removed from 326 great gerbils, two red foxes (Vulpes vulpes), three marbled polecats (Vormela peregusna), 35 pastured sheep (Ovis aries), and one long-eared desert hedgehog (Hemiechinus auritus) in the Gurbantunggut Desert. Ticks were identified according to standard morphological keys. Then, they were further analyzed by molecular and phylogenic methods based on two mitochondrial markers, 16S rDNA and cytochrome c oxidase subunit I (COI) genes. Results A total of 889 ticks were collected, representing five species. These included Hyalomma asiaticum (n = 425: 24 larvae, 79 nymphs and 322 adults), Rhipicephalus turanicus (n = 153: 2 nymphs and 151 adults), Haemaphysalis erinacei (n = 298: 4 larvae, 7 nymphs and 287 adults), Ixodes acuminatus (n = 7: 4 nymphs and 3 adults) and Ornithodoros tartakovskyi (6 adults). Based on COI sequences, molecular and phylogenetic analyses showed that (i) I. acuminatus from great gerbils and marbled polecats clustered with I. acuminatus reported from Europe; (ii) O. tartakovskyi found in northwestern China belonged to an independent clade; (iii) Hy. asiaticum, R. turanicus and Ha. erinacei had 100% sequence identities to conspecific ticks sampled previously in China. Conclusions The great gerbil is an important host for the developmental stages of I. acuminatus, O. tartakovskyi, Ha. erinacei, Hy. asiaticum and R. turanicus, thus supporting the life cycle of several tick species which, as adults, parasitize predators (red fox and marble polecat) as well as pastured sheep and hedgehogs in the Gurbantunggut Desert. Ixodes acuminatus and O. tartakovskyi were found for the first time on great gerbil and marbled polecat, respectively. Graphical Abstract

Disassembly line balancing (DLB) is a crucial optimization item in the recycling and remanufacturing of waste products. Considering the variations in the number of operators assigned to each station, this study investigates DLBs with six distinct station configurations: single-manned, multi-manned, single-robotic, multi-robotic, single-manned–robotic, and multi-manned–robotic setups. First, a unified mixed-integer programming (MIP) model is established for Type-I DLBs with each configuration to minimize four objectives: the number of stations, the number of operators, the total disassembly time, and the idle balancing index. To obtain more solutions, a novel bi-metric is proposed to replace the quadratic idle balancing index and is used in lexicographic optimization. Subsequently, based on the unified Type-I models, a unified MIP model for Type-II DLBs is established to minimize the cycle time, the number of operators, the total disassembly time, and the idle balancing index. Finally, the correctness of the established unified models and the effectiveness of the proposed bi-metric are verified by solving two disassembly cases of lighters and hairdryers, which further shows that the mathematical integration method of unified modeling has significant theoretical value for the multi-objective optimization of the DLBs with six distinct station configurations.

Equine piroplasmosis (EP) is an important tick-borne disease of equids, caused by Theileria equi, Theileria haneyi, and Babesia caballi. Nonetheless, there has been a scarcity of systematic reports on EP parasites in donkeys in Kyrgyzstan, Central Asia. In this study, piroplasms were screened in 1900 blood samples from imported donkeys from the Osh Oblast (southwestern Kyrgyzstan) by targeting partial 18S ribosomal RNA using the polymerase chain reaction (PCR). Through molecular and phylogenetic analyses, all positive samples were sequenced to identify the species and genotypes. The results indicated the presence of both B. caballi and T. equi, with prevalence rates of 8.4% (160/1900) and 12.2% (232/1900), respectively. By amplifying part of the Erythrocyte Merozoite Antigen 1 (EMA-1) and Rhoptry-Associated Protein (RAP-1) genes, B. caballi genotype B and T. equi genotype A were identified. To the best of our knowledge, this is the first report on piroplasm infection among donkeys from Kyrgyzstan.

Red foxes (Vulpes vulpes) have been recognized as natural reservoirs for multiple pathogens and a source of infection for domestic animals, wildlife and humans. To date, no reports are available on the Bartonella rochalimae and Hepatozoon canis infection in red foxes from China. In 2018–2022, a total of 16 red foxes were sampled in two counties and a city in Xinjiang Uygur Autonomous Region (XUAR) in northwest China. Subsequently analyzed by DNA extraction amplified by polymerase chain reaction (PCR). In the present study, based on nucleotide sequence and phylogenetic tree analyses, B. rochalimae and H. canis were molecularly identified in red foxes. Our findings provide the first molecular evidence of B. rochalimae and H. canis in red foxes from China.

Eurasian lynx (Lynx lynx) is widely distributed in various habitats in Asia and Europe, and it may harbor multiple pathogens. Currently, the information on protozoan infection in Eurasian lynx is scarce. In this study, we performed nested polymerase chain reaction (nPCR) analysis to detect intestinal protozoan infection in three dead Eurasian lynxes, in northwestern China. Three dead Eurasian lynxes, an adult female (#1), an adult male (#2), and a cub male (#3), were sampled in West Junggar Mountain, the northwestern region of Xinjiang Uyghur Autonomous Region. The intestine samples were analyzed using nPCR. We used primers targeting the cytochrome C oxidase subunit I gene (COI) for detection of Sarcocystis and Eimeria species and targeting the small subunit 18 S ribosomal RNA gene (18S rRNA) for detection of Cystoisospora species. The nPCR-positive products were sequenced, aligned, and phylogenetically analyzed. Three intestinal protozoa, Sarcocystis albifronsi, Eimeria alpacae, and Cystoisospora felis, were found in three Eurasian lynxes. The intestine sample of Eurasian lynx #2 was detected with S. albifronsi and E. alpacae. In addition, C. felis was only found in the intestine sample of Eurasian lynx #3. To the best of our knowledge, S. albifronsi and E. alpacae were detected in Eurasian lynx for the first time. In addition, C. felis was firstly found in Eurasian lynx in China. These findings extend our knowledge of the geographical distribution and host range of intestinal protozoa.

Human brucellosis caused by Brucella is a widespread zoonosis that is prevalent in many countries globally. The high homology between members of the Brucella genus and Ochrobactrum spp. often complicates the determination of disease etiology in patients. The efficient and reliable identification and distinction of Brucella are of primary interest for both medical surveillance and outbreak purposes. A large amount of genomic data for the Brucella genus was analyzed to uncover novel probes containing single-nucleotide polymorphisms (SNPs). GAMOSCE v1.0 software was developed based on the above novel eProbes. In conjunction with clinical requirements, an RPA-Cas12a detection method was developed for the on-site determination of B. abortus and B. melitensis by fluorescence and lateral flow dipsticks (LFDs). We demonstrated the potential of these probes for rapid and accurate detection of the Brucella genus and five significant Brucella species in silico using GAMOSCE. GAMOSCE was validated on different Brucella datasets and correctly identified all Brucella strains, demonstrating a strong discrimination ability. The RPA-Cas12a detection method showed good performance in detection in clinical blood samples and veterinary isolates. We provide both in silico and on-site methods that are convenient and reliable for use in local hospitals and public health programs for the detection of brucellosis.

Infections with spotted fever group rickettsiae represent a worldwide health problem, characterized by persistent high fever, headache, and rash in humans, domestic animals, and wildlife. To date, the occurrence of Rickettsia species in hard ticks has not been thoroughly studied, especially in eastern and southern Kazakhstan. A total of 1,245 adult ticks, comprising 734 Dermacentor marginatus, 219 Hyalomma scupense, 144 Hyalomma asiaticum, 84 Hyalomma marginatum, 48 Rhipicephalus turanicus, and 16 Haemaphysalis erinacei, collected from East Kazakhstan, Abay, Jetsu, Almaty, Jambyl, South Kazakhstan and Qyzylorda oblasts of Kazakhstan, were used to screen rickettsial agents using molecular methods. Rickettsia raoultii, Rickettsia slovaca, Rickettsia aeschlimannii and Rickettsia heilongjiangensis were identified using sequencing, and 31.5% (392/1245) of ticks carried rickettsial agents. The difference in the natural landscapes explains the variety of the collected ticks and expands our knowledge of Rickettsia species and their geographical distribution in Kazakhstan. To the best of our knowledge, this study reports the first finding of R. heilongjiangensis in Kazakhstan.

The role of gut microbiota has been described as an important influencer of the immune system. Gut-lung axis is critical in the prevention of mycobacterium infection, but the specific mechanism, by which dysbiosis affects tuberculosis, has not been reported. In this study, we attempted to provide more information on how the gut-lung axis contributes to Mycobacterium bovis (M. bovis) infection. Mice are pre-treated with broad-spectrum antibiotics cocktail (Abx) to induce gut dysbiosis. Interestingly, dysbiosis of microbes showed a significant increase in the bacterial burden in the lungs and inhibited the level of COX-2. After faecal transplantation, cyclooxygenase 2 (COX-2) expression was restored and the inflammatory lesion in the lungs was reduced. Further research found that the deficiency of COX-2 inhibited endoplasmic reticulum stress (ER stress). This mechanism was completed by COX-2 interaction with BIP. Moreover, we found a positive feedback mechanism by which blocking ER stress could reduce COX-2 levels by the NF-κB pathway. Taken together, we reveal for the first time gut dysbacteriosis exacerbates M. bovis disease by limiting the COX-2/ER stress pathway. The finding strengthens the foundation of gut microbiota-targeted therapy for tuberculosis treatment.

Respiratory mucosal immunization is an effective immunization strategy against tuberculosis (TB), and effective mucosal vaccines require adjuvants that can promote protective immunity without deleterious inflammation. Mucosal BCG (Bacille Calmette-Guerin) is effective, but it causes a severe inflammatory response in the lung. A novel less cytotoxic mucosal vaccine AH-PB containing Mycobacterium tuberculosis (Mtb) cell surface antigens Ag85A and HspX (AH), as well as polyinosinic-polycytidylic acid (Poly IC) and bovine neutrophil β-defensin-5 (B5) adjuvants were prepared, with the overarching goal of protecting against TB. Then, the immunogenicity and protective efficacy of these vaccines via the intranasal route were evaluated in a mouse model. Results showed that intranasal AH-PB promoted tissue-resident memory T cells (TRMs) development in the lung, induced antigen-specific antibody response in airway, provided protection against Mycobacterium bovis (M. bovis), conferred better protection than parenteral BCG in the later stage of infection, and boosted the protective immunity generated by BCG in mice. Moreover, both B5 and Poly IC were indispensable for the protection generated by AH-PB. Furthermore, intranasal immunization with AH-B5 fusion vaccines also provided similar protection against M. bovis compared to AH-PB. Collectively, B5-based TB vaccine via the intranasal route is a promising immunization strategy against bovine TB, and this kind of immunization strategy may be applied to human TB vaccine development. These findings highlight the potential importance of B5 as a mucosal adjuvant used in TB vaccines or other respiratory disease vaccines.

Abstract Background Among live wild mammals adapted to urban and peri-urban habitats in Europe, members of the families Felidae, Mustelidae and Sciuridae deserve special attention as pathogen reservoirs because all of these families include members that are kept as pets. We report here the results of our study on two important groups of tick-borne protozoan parasites in ticks and tissues of wild cats, mustelids and red squirrels. Methods DNA was extracted from the tissues of carnivores (wild cats, mustelids; n = 16) and red squirrels (n = 4), as well as from ixodid ticks (n = 89) collected from these hosts. These DNA extracts were screened for piroplasms and Hepatozoon spp. using conventional PCR analysis and sequencing. In addition, 53 pooled samples of 259 questing Haemaphysalis concinna ticks were evaluated for the presence of Hepatozoon DNA, followed by phylogenetic analyses. Results One wild cat was found to be coinfected with Cytauxzoon europaeus and a new genotype of Hepatozoon felis, and two additional wild cats were infected with H. felis from a different phylogenetic group. In mustelids, Hepatozoon martis and two further Hepatozoon genotypes were detected. The latter clustered separately, close to others reported from eastern Asia. In addition, Hepatozoon sciuri was detected in red squirrels. Morphologic and molecular analyses verified eight tick species. One wild cat was infected with a H. felis genotype that was significantly different from that in Ixodes ricinus females infesting this cat. Only three pools of questing H. concinna nymphs tested positive for Hepatozoon, one of which contained H. martis. Conclusions This study provides the first evidence of the occurrence of any Cytauxzoon species and of three Hepatozoon species in Hungary. In addition to H. martis, two further mustelid-associated Hepatozoon genotypes were detected, one of which was new in terms of phylogenetic and broader geographical contexts. This may be the first indication that H. felis genotypes from both of its phylogenetic groups occur in Europe. This also appears to be the first evidence of H. felis and C. europaeus coinfection in felids in Europe, and of autochthonous H. felis infection in wild cats north of the Mediterranean Basin. New tick–host associations were also observed in this study. Based on the results, H. felis and H. martis might survive transstadially in I. ricinus and H. concinna, respectively. Graphical Abstract

Accurately extracting buildings is essential for urbanization rate statistics, urban planning, resource allocation, etc. The high-resolution remote sensing images contain rich building information, which provides an important data source for building extraction. However, the extreme abundance of building types with large differences in size, as well as the extreme complexity of the background environment, result in the accurate extraction of spatial details of multi-scale buildings, which remains a difficult problem worth studying. To this end, this study selects the representative Xinjiang Tumxuk urban area as the study area. A building extraction network (SCA-Net) with feature highlighting, multi-scale sensing, and multi-level feature fusion is proposed, which includes Selective kernel spatial Feature Extraction (SFE), Contextual Information Aggregation (CIA), and Attentional Feature Fusion (AFF) modules. First, Selective kernel spatial Feature Extraction modules are used for cascading composition, highlighting information representation of features, and improving the feature extraction capability. Adding a Contextual Information Aggregation module enables the acquisition of multi-scale contextual information. The Attentional Feature Fusion module bridges the semantic gap between high-level and low-level features to achieve effective fusion between cross-level features. The classical U-Net, Segnet, Deeplab v3+, and HRNet v2 semantic segmentation models are compared on the self-built Tmsk and WHU building datasets. The experimental results show that the algorithm proposed in this paper can effectively extract multi-scale buildings in complex backgrounds with IoUs of 85.98% and 89.90% on the two datasets, respectively. SCA-Net is a suitable method for building extraction from high-resolution remote sensing images with good usability and generalization.

To address the current difficult problem of scientifically assessing the health status of date palm trees due to a single parameter for date palm health assessment, an imperfect index system, and low precision. In this paper, using jujube trees in 224 regiment of the 14th division of Xinjiang Production and Construction Corps “Kunyu city” as the research object, we carried out the inversion study of various physicochemical parameters of jujube trees (canopy chlorophyll content, leaf area index (LAI), tree height, canopy area) using the unmanned aerial vehicle (UAV) hyperspectral imagery of jujube trees during the period of fruit expansion, and put forward a model for assessing the health of jujube trees based on multiple physicochemical parameters. First, we calculated six spectral indices for inversion of chlorophyll content and four spectral index for inversion of LAI, analyzed the spectral index with high correlation with chlorophyll content and LAI of jujube trees canopy, and constructed the inversion models of chlorophyll content and LAI. Second, the Mask R-CNN model was used to achieve jujube trees’ canopy segmentation and area extraction, and the segmented canopy was matched with the Canopy Height Model (CHM) for jujube trees’ height extraction. Finally, based on the four physicochemical parameters of inversion, we construct four jujube trees’ health assessment models, namely, Partial Least Squares Regression Analysis (PLSR), Random Forest (RF), Support Vector Machines (SVM), and Decision Tree (DT). The results showed that the R2 of the PLSR tree health assessment model constructed based on the multi-physical and chemical parameters of chlorophyll content, LAI, tree height, and canopy area was 0.853, and the RMSE was 0.3. Compared with the jujube trees’ health assessment models constructed by RF, SVM, and DT, the R2 increased by 0.127, 0.386, and 0.165, and the RMSE decreased by 0.04, 0.175, and 0.063, respectively. This paper can achieve rapid and accurate inversion of multi-physical and chemical parameters of jujube trees with the help of UAV hyperspectral images, and the PLSR model constructed based on multi-physical and chemical parameters can accurately assess the health status of jujube trees and provide a reference for a scientific and reasonable assessment of jujube trees’ health.

The Eurasian lynx (Lynx) is a medium-sized wild cat species distributed throughout Eurasia. There has been no report on Taenia species (Cestoda: Cyclophyllidea) infecting this felid in China. In this study, 24 tapeworms were found in two Eurasian lynxes (#1 and #2) in Xinjiang Uygur Autonomous Region (XUAR), northwestern China. Based on the number, measurements and the shape of rostellar hooks, these tapeworms belong to two Taenia species. According to the number (n = 32) and length (185–194 μm) of small hooks, the first Taenia species (n = 1, found in #2 lynx) was identified as Taenia laticollis. Phylogenetically, this species was clustered with T. laticollis genotype C (JX860623) based on its cytochrome c oxidase subunit 1 (cox1) and 16S rDNA sequences. The second Taenia species (n = 23, provisionally named as “Taenia sp.“) may represent a potentially novel tapeworm species, because of its obvious differences in the shape and lengths (174–182 μm, 98–113 μm) of large and small rostellar hooks in comparison with ten taxonomically related species. Molecular and phylogenetic analyses of the cox1 gene revealed that “Taenia sp.” has the highest rate of sequence identity (92.93%, 368/396 bp) with Taenia hydatigena reported from sheep (Ovis aries) in Slovakia. To sum up, a potentially novel tapeworm species, “Taenia sp.“, is found in Eurasian lynx. In addition, T. laticollis was found for the first time in China.

In this study, 263 tapeworms were collected from eight road-killed red foxes in Xinjiang Uygur Autonomous Region (XUAR, northwestern China). The tapeworms were analyzed based on morphological characters and mitochondrial cytochrome c oxidase subunit 1 (cox1) gene sequences. Eighty-three Taenia and 180 Mesocestoides tapeworms were identified according to the presence or absence of rostellum, and the number, the length and the shape of the large rostellar hooks. The morphological and molecular analyses revealed that i) 180 Mesocestoides tapeworms, here named as Mesocestoides sp. (Vulpes vulpes), showed 99.21% (378/381 bp) identity to Mesocestoides sp. reported from red fox in Mongolia; and ii) 83 Taenia tapeworms belonged to three species. The first Taenia species [n = 16, named as Taenia sp. (Vulpes vulpes)], based on the length of large rostellar hooks (337–342 μm) and its cox1 sequence, was identified as a potentially novel species, which is phylogenetically close to Taenia laticollis. The second species [n = 54, named as Taenia sp. (Vulpes vulpes & Rhombomys opimus)], was morphologically similar to Taenia endothoracicus according to the number (n = 52), the length (319–332 μm) and the shape of the large rostellar hooks. This species, infecting three red foxes, shared 100% cox1 sequence identity with Taenia sp. (Rhombomys opimus) genotype C found previously in great gerbils (Rhombomys opimus) in the same region. The third species (n = 13, named as Taenia polyacantha-like), had shorter large rostellar hooks (178–180 μm) and showed 96.27% (361/375 bp) sequence identity to Taenia polyacantha reported from red fox in Italy. The “great gerbil-red fox” life cycle of Taenia sp. (Vulpes vulpes & Rhombomys opimus), belonging to the mitochondrial lineage of T. endothoracicus, is confirmed. The T. polyacantha-like species was firstly found in red fox in China. Taenia sp. (Vulpes vulpes) is a potentially novel species, which is close to T. laticollis based on its phylogenetic properties.

We aim to detect the presence of Tacheng tick virus 2 (TcTV-2) in ticks of southeastern Kazakhstan. A total of 205 ticks were collected and separated into four species, namely Hyalomma scupense, Dermacentor marginatus, Hyalomma asiaticum and Hyalomma anatolicum. The partial S segment of TcTV-2 was detected in individual RNA of separated ticks by reverse transcriptase polymerase chain reaction. 11.22% (23/205) of ticks were positive to the viral S segment. This is first report of presence of the TcTV-2 in Hy. scupense and Hy. anatolicum from Kazakhstan. This finding extends tick species and the geographic distribution of TcTV-2.

Abstract Background Increasing numbers of tick-borne pathogens are being discovered, including those that infect humans. However, reports on co-infections caused by two or more tick-borne pathogens are scarce. Case presentation A 38-year-old male farmer was bitten by a hard tick, presented with fever (37.7 °C), severe headache and ejection vomiting. Lumbar puncture was performed in the lateral decubitus. The cerebrospinal fluid (CSF) was clear, and analysis showed severe increased pressure (320 mm H2O), mild leukocytosis (126.0 × 106/L, mononuclear cells accounting for 73%) and elevated total protein concentration (0.92 g/L). Bacterial cultures of CSF and blood were negative. The diagnosis of Rickettsia raoultii and Tacheng tick virus 1 (TcTV-1) co-infection was confirmed by amplifying four rickettsial genetic markers and the partial small (S) RNA segment of TcTV-1 from the patient’s blood. The patient gradually recovered after treatment with levofloxacin and ribavirin. Conclusions This is the first reported co-infection case with fever and meningitis caused by R. raoultii and TcTV-1. It is vital to screen for multiple pathogens in tick-bitten patients, especially in those with severe complex symptoms.

The great gerbil (Rhombomys opimus), widely distributed in Asia, is a natural reservoir for Yersinia pestis, Leishmania donovani and some species of helminths. In this study, 188 great gerbils were sampled in Alataw City and Manas County, northwestern China, and tested for the presence of Theileria, Hepatozoon and Taenia species by molecular methods. Theileria sp., named as “Candidatus Theileria xinjiangensis”, was detected in heart, liver, spleen, lung, and kidney of 6.9% rodents. Six genotypes of “Taenia sp. Rhombomys opimus”, which were close to Taenia laticollis (87.3–94.0% identities), were detected in cyst liquid of 5.3% rodents. “Hepatozoon ayorgbor-like” haemogregarines was detected in spleens of 1.6% rodents. To our best knowledge, Candidatus Theileria xinjiangensis, Hepatozoon ayorgbor-like and genotypes of “Taenia sp. Rhombomys opimus” were found for the first time in the great gerbil. These results extend our knowledge on the diversity and pathogenesis of Theileria, Hepatozoon and Taenia species.

Abstract Background This case report describes the clinical process of a shepherd who suffered brucellosis-related endocarditis (BE) and spondylitis (BS) and was infected with Brucella melitensis biovar 3 (B. melitensis biovar 3). Case presentation A 55-year-old male patient was admitted to The First Affiliated Hospital of Shihezi University on October 11, 2018, due to over 3 months of intermittent fever, back pain, and heart trouble. The Rose Bengal Plate test was positive, the standard agglutination test titer for brucellosis was 1/800, and the blood culture was positive for B. melitensis biovar 3. Three instances of transthoracic echocardiography examination at days 1, 25, and 376 after admission to the hospital and magnetic resonance imaging (MRI) and computed tomography (CT) checks at days 5 and 38 revealed that the size of the vegetation on the posterior leaflet of the mitral valve increased from 0.7 × 1.4 cm to 1.2 × 1.5 cm and that the left atrium and ventricle were enlarged. The MRI and CT results showed hyperplasia of the second and third vertebra, a cold abscess formed on both sides of the psoas major muscles, and the vertebra hyperplasia became aggravated at a later time point. The patient’s situation deteriorated, and heart failure was discovered on October 22, 2019. At the moment of submission of this manuscript, the patient remains in bed at home because of severe debility caused by brucellosis. Conclusions This is the first reported case of endocarditis combined with spondylitis caused by B. melitensis biovar 3 in a shepherd. Brucellosis infection can cause work-power losses because of misdiagnosis or a lack of proper treatment. Early diagnosis and treatment are essential for a successful outcome.

Decompressive craniectomy (DC) is of great significance for relieving acute intracranial hypertension and saving lives after traumatic brain injury (TBI). In this study, a severe TBI mouse model was created using controlled cortical impact (CCI), and a surgical model of DC was established. Furthermore, a series of neurological function assessments were performed to better understand the pathophysiological changes after DC. In this study, mice were randomly allocated into three groups, namely, CCI group, CCI+DC group, and Sham group. The mice in the CCI and CCI+DC groups received CCI after opening a bone window, and after brain injury, immediately returned the bone window to simulate skull condition after a TBI. The CCI+DC group underwent DC and contused tissue removal 6 h after CCI. The mice in the CCI group underwent the same anesthesia process; however, no further treatment of the bone window and trauma was performed. The mice in the Sham group underwent anesthesia and the process of opening the skin and bone window, but not in the CCI group. Changes in Modified Neurological Severity Score, rotarod performance, Morris water maze, intracranial pressure (ICP), cerebral blood flow (CBF), brain edema, blood–brain barrier (BBB), inflammatory factors, neuronal apoptosis, and glial cell expression were evaluated. Compared with the CCI group, the CCI+DC group had significantly lower ICP, superior neurological and motor function at 24 h after injury, and less severe BBB damage after injury. Most inflammatory cytokine expressions and the number of apoptotic cells in the brain tissue of mice in the CCI+DC group were lower than in the CCI group at 3 days after injury, with markedly reduced astrocyte and microglia expression. However, the degree of brain edema in the CCI+DC group was greater than in the CCI group, and neurological and motor functions, as well as spatial cognitive and learning ability, were significantly poorer at 14 days after injury.

Despite the recognized epidemiological importance of ticks as vectors for pathogens that cause numerous zoonotic and veterinary diseases, data regarding the pathogens of pet dogs and their parasitic ticks in the Junggar Basin are scarce. In this study, a total of 178 blood samples and 436 parasitic ticks were collected from pet dogs in Junggar Basin, Xinjiang Uygur Autonomous Region (XUAR), north-western China. All ticks were identified as Rhipicephalus turanicus sensu stricto (s.s.) according to morphological and molecular characteristics. Rh. turanicus s.s. ticks were collected from pet dogs in China for the first time. Seven tick-borne pathogens, such as Ehrlichia chaffeensis, Anaplasma phagocytophilum, Rickettsia massiliae, Candidatus R. barbariae, Brucella spp., Rickettsia sibirica, and Anaplasma ovis, were detected from ticks, whereas the first five bacteria were detected from blood samples of dogs. Brucella spp. was the most predominant pathogen in both blood samples and ticks of pet dogs, with the detection rates of 16.29 and 16.74%, respectively. Moreover, 17 ticks and 1 blood sample were co-infected with two pathogens, and 1 tick was co-infected with three pathogens. This study provided molecular evidence for the occurrence of Anaplasma spp., Ehrlichia spp., Rickettsia spp., and Brucella spp. circulating in pet dogs and their parasitic ticks in Junggar Basin, north-western China. These findings extend our knowledge of the tick-borne pathogens in pet dogs and their parasitic ticks in Central Asia; therefore, further research on these pathogens and their role in human and animal diseases is required.

Brucellosis, caused by Brucella spp., is one of the most widespread bacterial zoonoses worldwide. Vaccination is still considered the best way to control brucellosis. An investigation into the differential proteome expression patterns of wild and vaccine strains may help researchers and clinicians differentiate between the strains to diagnose and better understand the mechanism(s) underlying differences in virulence. In the present study, a mass spectrometry-based, label-free relative quantitative proteomics approach was used to investigate the proteins expressed by the wild strain, B. melitensis biovar 3 and compare it with those expressed by B. melitensis M5-90. The higher level of virulence for B. melitensis biovar 3 compared to B. melitensis M5-90 was validated in vitro and in vivo. A total of 2133 proteins, encompassing 68% of the theoretical proteome, were identified and quantified by proteomic analysis, resulting in broad coverage of the B. melitensis proteome. A total of 147 proteins were identified as differentially expressed (DE) between these two strains. In addition, 9 proteins and 30 proteins were identified as unique to B. melitensis M5-90 and B. melitensis biovar 3, respectively. Pathway analysis revealed that the majority of the DE proteins were involved in iron uptake, quorum sensing, pyrimidine metabolism, glycine betaine biosynthetic and metabolic processes, thiamine-containing compound metabolism and ABC transporters. The expression of BtpA and VjbR proteins (two well-known virulence factors) in B. melitensis biovar 3 was 8-fold and 2-fold higher than in B. melitensis M5-90. In summary, our results identified many unique proteins that could be selected as candidate markers for differentiating vaccinated animals from animals with wild-type infections. BtpA and VjbR proteins might be responsible for the residual virulence of B. melitensis M5-90, while ABC transporters and thiamine metabolism associated proteins may be newly identified Brucella virulence factors. All of the identified DE proteins provide valuable information for the development of vaccines and the discovery of novel therapeutic targets.

Abstract Background Previously, twelve Rickettsia species were identified in ticks, fleas, sheep keds (Melophagus ovinus), bats (Pipistrellus pipistrellus) and a tick-bitten patient in the Xinjiang Uygur Autonomous Region (XUAR) in northwestern China. Here we aimed to molecularly detect rickettsial agents in red fox (Vulpes vulpes), marbled polecat (Vormela peregusna) and their ticks. Methods During 2018–2019, 12 red foxes, one marbled polecat and their ticks were sampled in two counties and a city of the XUAR. The heart, liver, spleen, lung and kidney of these 13 carnivores were dissected, followed by DNA extraction. Hard ticks were identified both morphologically and molecularly. All samples were examined for the presence of rickettsiae by amplifying four genetic markers (17-kDa, gltA, ompA, sca1). Results A total of 26 adult ticks and 28 nymphs (38 Ixodes canisuga, nine Ixodes kaiseri, six Haemaphysalis erinacei and one Dermacentor marginatus) were collected from red foxes, and four Ha. erinacei ticks were removed from the marbled polecat. Analysis of cytochrome c oxidase subunit I (COI) gene sequences indicated that 2–32 nucleotides differed between I. canisuga, I. kaiseri and Ha. erinacei from northwestern China and Europe. Rickettsia raoultii was detected in three red foxes, Candidatus Rickettsia barbariae in a red fox, Rickettsia sibirica in a red fox and a marbled polecat, and R. raoultii in two tick species (I. canisuga and D. marginatus). Conclusions To the best of our knowledge, I. canisuga and I. kaiseri have not been previously reported from red foxes in China. The DNA of R. sibirica and R. raoultii was detected for the first time in the organs of red foxes, and R. sibirica in the organs of a marbled polecat. This is also the first molecular evidence for the presence of R. raoultii in I. canisuga. Our findings expand the range of tick-borne pathogens in wildlife species and associated ticks in China.

We used metagenomic analysis to identify Tacheng tick virus 2 infection in a patient with a history of tick bite in northwestern China. We confirmed the virus with reverse transcription-PCR, virus isolation, and genomic analysis. We detected viral RNA in 9.6% of ticks collected from the same region.

The wet deposition flux of black carbon (BC) over the Sichuan Basin is studied on the basis of the MERRA-2 data from 1981 to 2020, aiming at investigating high BC wet deposition flux in China in terms of long-term spatial-temporal trends and influences of BC column mass density and precipitation. In China, the largest BC wet deposition flux with a regionally-averaged value of 1.00 × 10−2 μg m−2 s−1 over the Sichuan Basin is observed, especially in the western and southern regions of the Basin with values as high as 2.20 × 10−2 μg m−2 s−1. The seasonality of BC wet deposition flux over the Sichuan Basin depicts maximum levels in autumn, moderate levels in spring and winter, and minimum levels in summer. The monthly mean BC wet deposition flux varies almost twofold, ranging from the lowest average value of 8.05 × 10−3 μg m−2 s−1 in July to the highest 1.28 × 10−2 μg m−2 s−1 in October. This study suggests that BC column mass density and precipitation are two significant factors affecting high BC wet deposition flux, whereas BC wet deposition flux is more related to BC column mass density than to precipitation over the Sichuan Basin.

Brucella utilizes diverse virulence factors to modulate the infectious cycle and lifestyle associated with eukaryotic hosts. Lipopolysaccharides (LPS) play an important role in the establishment of persistent infections of Brucella, but its mechanism in host cell to interfere with the host"s specific signaling pathway related to the elimination of Brucella is still not clear. LpsA, a glycosyl transferase, involves in the biosynthesis of LPS, and may aff ect the intracellular survival of Brucella. This study aimed to investigate the eff ects of lpsA on pyroptosis in Brucella-infected macrophage cells. We constructed the lpsA mutant strain (M5-90ΔlpsA) and the complementary strain (M5-90ΔlpsA-C) of Brucella melitensis strain M5-90 infected mouse macrophages to detect pyroptosis. We found that the inactivation of the lpsA gene weakened the ability of LPS gains access to cytosol during Brucella mutant infections, and reduced the survival of Brucella. Western blot and quantitative real-time PCR assays showed that the mRNA and protein levels of caspase-11 and NOD-like receptor family pyrin domain-containing 3 in M5-90ΔlpsA group were higher than those observed in M5-90 group. These results indicated M5-90ΔlpsA promoted the pyroptosis of RAW264.7 cells (a mouse macrophage cell line) after Brucella infection. The finding reveals that Brucella lpsA plays a partial role in innate immunity and infl ammatory response by inhibiting the LPS-induced atypical pyroptosis pathway.

Bacterial load in clinical samples is relatively low and difficult to detect. Improvements in assay sensitivity will greatly reduce false negative results and contribute to more accurate diagnoses. In the present study, we present a new strategy to improve the sensitivity of a nucleic acid assay by detecting the presence of a multi-copy gene. By using Brucella as a test model, we screened the genome and identified IS711 as a multiple copy gene. Distribution analysis of insertion sequence IS711 among different species and strains showed that each of the strains have 5 to 13 copies of IS711. Compared with the BMEI1001, BMEI0775 and BMEI0027, the assays of high copy genes IS711 showed higher sensitivity and is an ideal high copy signature gene for Brucella. Detection of clinical samples with assays targeting the signature genes showed that IS711 exist in higher concentrations than BMEI1001, BMEI0775 and BMEI0027. In addition, IS711 assay is more sensitive than other signature genes assay. Analysis of several other pathogenic bacteria successfully identified high copy number genes that could be used as signature genes. Therefore, this strategy of targeting high copy signature genes represents a universal strategy for the ultrasensitive detection of bacteria.

Yersinia pestis, the cause of plague, is a newly evolved Gram-negative bacterium. Through the acquisition of the plasminogen activator (Pla), Y. pestis gained the means to rapidly disseminate throughout its mammalian hosts. It was suggested that Y. pestis utilizes Pla to interact with the DEC-205 (CD205) receptor on antigen-presenting cells (APCs) to initiate host dissemination and infection. However, the evolutionary origin of Pla has not been fully elucidated. The PgtE enzyme of Salmonella enterica, involved in host dissemination, shows sequence similarity with the Y. pestis Pla. In this study, we demonstrated that both Escherichia coli K-12 and Y. pestis bacteria expressing the PgtE-protein were able to interact with primary alveolar macrophages and DEC-205-transfected CHO cells. The interaction between PgtE-expressing bacteria and DEC-205-expressing transfectants could be inhibited by the application of an anti-DEC-205 antibody. Moreover, PgtE-expressing Y. pestis partially re-gained the ability to promote host dissemination and infection. In conclusion, the DEC-205-PgtE interaction plays a role in promoting the dissemination and infection of Y. pestis, suggesting that Pla and the PgtE of S. enterica might share a common evolutionary origin.

Twenty-five dog fleas (Ctenocephalides canis) and five ticks identified as Hyalomma asiaticum were collected from a Eurasian lynx (Lynx lynx) pup in northwestern China. Molecular analyses of four genetic markers showed the presence of Rickettsia aeschlimannii DNA in 5 out of 20 fleas. Only rickettsial 17-kDa gene was detected in the blood sample of the lynx. In addition, 2 out of 20 fleas were positive to Wolbachia endosymbiont by targeting 16S rDNA while there was no Bartonella DNA found both in 5 ticks and 20 fleas by using gltA and 16S-23S ITS. Our findings suggest that i) C. canis parasitizing wild Eurasian lynx harbors R. aeschlimannii and Wolbachia endosymbiont in the China-Kazakhstan border, and ii) Wolbachia endosymbiont in present study is closer to that in C. canis infesting sheltered dogs in Turkey.

Flea distribution in northern region of Xinjiang Uygur Autonomous Region (XUAR) and fluctuations of the annual fleas index in Alataw Pass were investigated. During a 4-year (2015–2018) study, 5789 fleas were collected directly from 15 mammals at eight counties in northern XUAR of northwestern China. Nineteen flea species, belonging to sixteen genera and seven families, were further confirmed by four genetic markers (18S rDNA, 28S rDNA, COI and COII) after morphological observation. Pulex irritans and Paraceras crispum parasitizing Asian badgers (Meles leucurus) were recorded for the first time. In addition, the fluctuations of the annual fleas index in Alataw Pass were surveyed. Xenopsylla gerbilli minax, Xenopsylla conformis conformis and Nosopsyllus laeviceps laeviceps were highly detected in the warm season while Paradoxopsyllus repandus, Ctenophthalmus dolichus dolichus and Coptopsylla lamellifer ardua were only found in the cold season. These findings extend our knowledge of flea species, distribution and annual fluctuations especially in China-Kazakhstan border. Keywords: Flea distribution, XUAR, Fluctuations, The annual fleas index, Wild mammals

Abstract Background Increasing molecular evidence supports that bats and/or their ectoparasites may harbor vector-borne bacteria, such as bartonellae and borreliae. However, the simultaneous occurrence of rickettsiae in bats and bat ticks has been poorly studied. Methods In this study, 54 bat carcasses and their infesting soft ticks (n = 67) were collected in Shihezi City, northwestern China. The heart, liver, spleen, lung, kidney, small intestine and large intestine of bats were dissected, followed by DNA extraction. Soft ticks were identified both morphologically and molecularly. All samples were examined for the presence of rickettsiae by amplifying four genetic markers (17-kDa, gltA, ompA and ompB). Results All bats were identified as Pipistrellus pipistrellus, and their ticks as Argas vespertilionis. Molecular analyses showed that DNA of Rickettsia parkeri, R. lusitaniae, R. slovaca and R. raoultii was present in bat organs/tissues. In addition, nine of the 67 bat soft ticks (13.43%) were positive for R. raoultii (n = 5) and R. rickettsii (n = 4). In the phylogenetic analysis, these bat-associated rickettsiae clustered together with conspecific sequences reported from other host and tick species, confirming the above results. Conclusions To the best of our knowledge, DNA of R. parkeri, R. slovaca and R. raoultii was detected for the first time in bat organs/tissues. This is also the first molecular evidence for the presence of R. raoultii and R. rickettsii in bat ticks. To our knowledge, R. parkeri was not known to occur in Asia. Our results highlight the need to assess rickettsial agents in a broader range of bat species and associated tick species.

This paper presents a piezoresistive differential pressure sensor based on a silicon-on-insulator (SOI) structure for low pressure detection from 0 to 30 kPa. In the design phase, the stress distribution on the sensing membrane surface is simulated, and the doping concentration and geometry of the piezoresistor are evaluated. By optimizing the process, the realization of the pressure sensing diaphragm with a controllable thickness is achieved, and good ohmic contact is ensured. To obtain higher sensitivity and high temperature stability, an SOI structure with a 1.5 µm ultra-thin monocrystalline silicon layer is used in device manufacturing. The device diaphragm size is 700 µm × 700 µm × 2.1 µm. The experimental results show that the fabricated piezoresistive pressure sensor has a high sensitivity of 2.255 mV/V/kPa and a sensing resolution of less than 100 Pa at room temperature. The sensor has a temperature coefficient of sensitivity (TCS) of −0.221 %FS/°C and a temperature coefficient of offset (TCO) of −0.209 %FS/°C at operating temperatures ranging from 20 °C to 160 °C. The reported piezoresistive microelectromechanical systems (MEMS) pressure sensors are fabricated on 8-inch wafers using standard CMOS-compatible processes, which provides a volume solution for embedded integrated precision detection applications of air pressure, offering better insights for high-temperature and miniaturized low-pressure sensor research.

Hybrid systems that integrate wave energy converters (WECs) with floating offshore wind turbines (FOWTs) are considered to be key equipment to deeply exploit marine renewable energy. The power take-off (PTO) system is an important component of the hybrid system, whose parameters also have a significant impact on the hybrid system’s performance. In this paper, a wind-wave hybrid system using hydraulic PTO systems is proposed. A numerical simulation framework based on the linear wave theory and basic equations of hydraulic components is built and verified. The influence of six critical hydraulic parameters on the wave energy capture and motion response performance of the hybrid system is investigated. Specifically, the parameters of piston area, motor displacement, and equivalent generator damping affect the performance of the hybrid system similar to changing the damping term of the PTO system. The parameters of the initial gas volume and the pre-charged pressure of the accumulator affect the wave power capture only for short wave periods, while the motion response of the hybrid system increases with the increase of these two parameters. The parameter of orifice area of the throttle valve affects the performance of the hybrid system slightly only when it is small. The optimal value of partial hydraulic parameters and their corresponding peak performance are also discussed.

It has been established that kallikrein12 (KLK12) expression is closely related to bovine tuberculosis (bTB) development. Herein, we sought to clarify the regulatory mechanism of KLK12 and its application in tuberculosis diagnosis. KLK12 knockdown macrophages were produced by siRNA transfection. Bradykinin receptors (BR, including B1R and B2R) were blocked with specific inhibitors. Mannose-capped lipoarabinomannan (ManLAM) was extracted from Mycobacterium bovis (M. bovis) and used to study the mechanism of KLK12 activation. In addition, we constructed different mouse models representing the latent and active stages of M. bovis infection. Mouse models and clinical serum samples were used to assess the diagnostic value of biomarkers. Through the above methods, we confirmed that KLK12 regulates MMP-1 and MMP-9 via BR. KLK12 upregulation is mediated by the M. bovis-specific antigen ManLAM. KLK12, MMP-1, and MMP-9 harbor significant value as serological markers for differentiating between latent and active bTB, especially KLK12. In conclusion, we identified a novel signaling pathway, KLK12/BR/ERK/MMPs, in M. bovis-infected macrophages, which is activated by ManLAM. From this signaling pathway, KLK12 can be used as a serological marker to differentiate between latent and active bTB. Importantly, KLK12 also has enormous potential for the clinical diagnosis of human tuberculosis (TB).

We isolated Brucella melitensis biovar 3 from the spleen of an Asian badger (Meles leucurus) in Nilka County, northwestern China. Our investigation showed that this isolate had a common multilocus variable-number tandem-repeat analysis 16 genotype, similar to bacterial isolates from local aborted sheep fetuses.

The zoonotic rickettsial pathogen Anaplasma species have a broad geographical distribution and are important intracellular agents. Domestic animals and wild rodents may play an important role in the epidemiology of this disease. The aim of this study was to estimate the prevalence of infection with Anaplasma species among domestic animals and wild rodents in northern Xinjiang Uygur Autonomous Region (XUAR), Northwest China, during 2015-2016. In this study, DNA from Anaplasma ovis was detected by nested PCR in blood samples from 21/137 sheep (15.3%) in Ili Kazakh Autonomous Prefecture (Ili), 18/79 sheep (22.8%) in Bole Mongol Autonomous Prefecture (Bole) and 13/71 sheep (18.3%) in Shihezi City. In addition, detection of Anaplasma phagocytophilum DNA in spleen samples from great gerbils (Rhombomys opimus) in Urumqi (37/356, 10.4%) and Bole (21/134, 15.7%). Interestingly, co-infection A. ovis and A. phagocytophilum in blood of sheep (9/137, 6.7%) from Ili, northern XUAR. Based on 16S rRNA sequence, phylogenetic analyses showed that A. ovis were separated into one clade, while the A. phagocytophilum was separated into another clade. This analysis demonstrated that there are at least two different Anaplasma species widespread. The present investigation revealed high infection rates of A. phagocytophilum and A. ovis, which shed light on making effective measures to prevent and control Anaplasma species infection in animals in XUAR, Northwest China.

Abstract Background The number of new rickettsial species are rapidly increasing, and increasing numbers of Rickettsia raoultii (R. raoultii) infection cases have been detected in humans. However, neurological abnormalities caused by R. raoultii are rarely reported, especially in northwestern China. Case presentation A 36-year-old Kazakh shepherd with an attached tick on part temporalis, presented with right eyelid droop, lethargy, fever, headache, fever (38.0–41.0 °C) and erythematous rash. The examination of cerebrospinal fluid (CSF) showed cerebrospinal pressure of 200 mm H2O, leukocyte count of 300.0 × 106/L, adenosine deaminase of 2.15 U/L, and total protein concentration of 0.93 g/L. The diagnosis of R. raoultii infection was confirmed by six genetic markers, and semi-quantified by enzyme-linked immunosorbent assay for rickettsial antigen. The patient gradually recovered after treatment with doxycycline and ceftriaxone. R. raoultii DNA was found both in a tick detached from this patient and in 0.18% (2/1107) of blood samples collected from local shepherds. Conclusions This is the first reported case with neurological abnormalities caused by R. raoultii in northwestern China. It is vital to detect rickettsial agents both in blood and CSF for tick bite patients with neurological abnormalities. Public health workers and physicians should pay attention to neurological abnormalities caused by Rickettsia.

This study aimed to investigate the cause of abortion in a traditional sheep farm. Here, 31 specimens were examined by PCR targeting the Brucella outer membrane protein gene 22 (omp22), and 25 (80.6%, 25/31) were found to be positive. Totally, 3 Brucella suis S2 and 10 Brucella melitensis were isolated from 31 aborted fetuses in which one B. suis S2 and B. melitensis were isolated from a same aborted fetus. All of these isolates were identified by PCR and conventional bacterial tests. These 3 B. suis isolates and the reference strain B. suis S2 were all identified as B. suis biovar 1 and the 10 B. melitensis isolates were all identified as B. melitensis biovar 3. This study suggests that B. suis S2 can partially induce abortion in pregnant ewes. Hence, for the sake of safety, it is need to develop a new Brucella vaccine to protect pregnant sheep from brucellosis.

Abstract Background Hantaviruses are important zoonotic pathogens, and they pose a profound risk to public health. So far, there has been no evidence showing that Tula virus (TULV), one species of hantavirus, is endemic in China. In this study, we captured rodents and found that the Tula virus had infected voles in Yili region, Xinjiang, China. Methods Rodents were captured by flooding their burrows in mountain pasture areas in Narati, Xinyuan County, Xinjiang, China. Hantavirus L gene fragments were amplified by nest RT-PCR using genus-specific primers. Positive samples were further identified by sequencing of RT-PCR products of S gene fragment for species identification. To identify the species of captured small mammals, the rodents’ cytochrome b (Cytb) was amplified by PCR and sequenced. Phylogenetic analysis was used to show the clustering and evolution relationship of the viral nucleic acids. Results Here, 31 out of 198 voles captured (16%) were infected with TULV. Host sequencing analysis showed these voles were Microtus obscurus (M. obscurs). Alignment and phylogenetic analysis of the exon region (1191 bp) of the hantavirus S gene confirmed that all of the detected amplicons were TULV, which was similar to one strain of TULV identified in Kazakhstan. Conclusion This is the first identification of Tula virus in China, and we found that M. obscurus acts as a natural reservoir for carrying the virus. Although the infection rate in the local human population remains unknown, the high prevalence of TULV in the small mammals in the region constitutes a risk that this putative pathogen may spread to the local population.

Abstract Background Despite the increasingly recognized eco-epidemiological significance of bats, data from molecular analyses of vector-borne bacteria in bat ectoparasites are lacking from several regions of the Old and New Worlds. Methods During this study, six species of ticks (630 specimens) were collected from bats in Hungary, Romania, Italy, Kenya, South Africa, China, Vietnam and Mexico. DNA was extracted from these ticks and analyzed for vector-borne bacteria with real-time PCRs (screening), as well as conventional PCRs and sequencing (for pathogen identification), based on the amplification of various genetic markers. Results In the screening assays, Rickettsia DNA was only detected in bat soft ticks, whereas Anaplasma phagocytophilum and haemoplasma DNA were present exclusively in hard ticks. Bartonella DNA was significantly more frequently amplified from hard ticks than from soft ticks of bats. In addition to Rickettsia helvetica detected by a species-specific PCR, sequencing identified four Rickettsia species in soft ticks, including a Rickettsia africae-like genotype (in association with a bat species, which is not known to migrate to Africa), three haemotropic Mycoplasma genotypes in Ixodes simplex, and Bartonella genotypes in I. ariadnae and I. vespertilionis. Conclusions Rickettsiae (from both the spotted fever and the R. felis groups) appear to be associated with soft rather than hard ticks of bats, as opposed to bartonellae. Two tick-borne zoonotic pathogens (R. helvetica and A. phagocytophilum) have been detected for the first time in bat ticks. The present findings add Asia (China) to the geographical range of R. lusitaniae, as well as indicate the occurrence of R. hoogstraalii in South Africa. This is also the first molecular evidence for the autochthonous occurrence of a R. africae-like genotype in Europe. Bat haemoplasmas, which are closely related to haemoplasmas previously identified in bats in Spain and to “Candidatus Mycoplasma haemohominis”, are reported here for the first time from Central Europe and from any bat tick.