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Background: Ticks are widely distributed throughout China and are the second most prevalent pathogen vectors in the world, following only mosquitoes. Tick bites can lead to Lyme disease, forest encephalitis, and other illnesses that may result in death under severe circumstances. Materials and methods: Ticks collected from March 2021 to May 2023 were pooled and used in metatranscriptomic analyses to gain insight into the diversity and distribution of tick-borne viruses in Inner Mongolia. Next-generation sequencing (NGS) outcomes were validated, and viral prevalence across distinct tick species was determined through the application of polymerase chain reaction (PCR) paired with Sanger sequencing. Results: A total of 20 RNA viruses belonging to at least 8 families, including Chuviridae, Flaviviridae, Solemoviridae, Nairoviridae, Partitiviridae, Phenuiviridae, Rhabdoviridae, and Totiviridae, and to unclassified families were identified by NGS. Five of the identified RNA viruses (Nuomin virus, Yezo virus, tick-borne encephalitis virus, Alongshan virus, and Beiji nairovirus) are considered human pathogens. A potential human pathogen, Mukawa virus, was also among the identified viruses. Ixodes persulcatus carried a significantly greater number of viral species than did Dermacentor nuttalli, Hyalomma marginatum, and Haemaphysalis concinna. The prevalence of coinfection with multiple viruses differed in I. persulcatus from Hinggan League and Hulun Buir, and Beiji nairovirus was the codominant virus species. Conclusions: There is a remarkable diversity of RNA viruses harboured by ticks in Inner Mongolia, with variations observed in the distribution of these tick-borne viruses across different regions and tick hosts. Author summary: Ticks are important vectors of zoonotic diseases and play a major role in the circulation and transmission of such diseases. In this study, ticks collected from March 2021 to May 2023 were pooled and subjected to metatranscriptomic analysis to gain insight into the diversity and distribution of tick-borne viruses in Inner Mongolia. Among the identified RNA viruses, 6 are recognized as human pathogens or potential human pathogens; these are Nuomin virus, Yezo virus, tick-borne encephalitis virus, Alongshan virus, Beiji nairovirus, and Mukawa phlebovirus. This study revealed significant diversity among RNA viruses transported by ticks in Inner Mongolia as well as variations in their geographic distribution. To effectively detect alterations in pathogen distribution, ongoing surveillance is urgently needed. [ABSTRACT FROM AUTHOR]

Ticks are important vectors of zoonotic diseases and play a major role in the circulation and transmission of many rickettsial species. The aim of this study was to investigate the carriage of Candidatus Rickettsia tarasevichiae (CRT) in a total of 1168 ticks collected in Inner Mongolia to elucidate the potential public health risk of this pathogen, provide a basis for infectious disease prevention, control and prediction and contribute diagnostic ideas for clinical diseases that present with fever in populations exposed to ticks. A total of four tick species, Haemaphysalis concinna (n = 21), Dermacentor nuttalli (n = 122), Hyalomma marginatum (n = 148), and Ixodes persulcatus (n = 877), were collected at nine sampling sites in Inner Mongolia, China, and identified by morphological and molecular biological methods. Reverse transcription PCR targeting the 16S ribosomal RNA (rrs), gltA, groEL, ompB and Sca4 genes was used to detect CRT DNA. Sequencing was used for pathogen species confirmation. The molecular epidemiological analysis showed that three species of ticks were infected with CRT, and the overall positive rate was as high as 42%. The positive rate of I. persulcatus collected in Hinggan League city was up to 96%, and that of I. persulcatus collected in Hulun Buir city was 50%. The pool positive rates of D. nuttalli and H. marginatum collected in Bayan Nur city and H. concinna collected in Hulun Buir city were 0%, 28% and 40%, respectively. This study revealed the high prevalence of CRT infection in ticks from Inner Mongolia and the first confirmation of CRT detected in H. marginatum in China. The wide host range and high infection rate in Inner Mongolia may dramatically increase the exposure of CRT to humans and other vertebrates. The role of H. marginatum in the transmission of rickettsiosis and its potential risk to public health should be further considered. [ABSTRACT FROM AUTHOR]

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Spotted fever group (SFG) rickettsioses are important zoonoses, threatening human health seriously and gradually attracting more attention in the world. SFG rickettsiae are classified as neglected pathogens. If these pathogens are detected at all, they are usually recognized very late in the infection through indirect detection of specific antibodies. Previous studies have shown that Rickettsia raoultii (R. raoultii), a member of the SFG rickettsiae, occurs with increasing incidence in remote countries. Therefore, a rapid detection method for R. raoultii is in urgently need. In this study, a R. raoultii diagnosis method by closed dumbbell-mediated isothermal amplification (R-CDA) assay targeting a conserved sequence of the outer membrane protein A (OmpA) gene with high sensitivity and specificity was developed. This assay offered a rapid and simple method for on-site detection of R. raoultii. Firstly, four pairs of R-CDA primers were designed and the optimum primer set was selected to amplify target gene specifically and effectively. Then, a pair of outer primer was designed to accelerate the reaction based on the inner primers to establish the RO-CDA reaction. In addition, the results of real-time amplification curves, melting curves and end-point colorimetric judgements showed that the established visual RO-CDA reaction could accurately detect R. raoultii without cross-reaction with other closely related pathogens. Furthermore, the detection limit of visual RO-CDA assay was 10 copies/μL, which was feasible for on-site detection with merits of easy-operation, rapidity, high sensitivity, and specificity. In conclusion, the developed RO-CDA detection method could be helpful for pathogen screening and epidemic prevention at the point of care. Author summary: Spotted fever group (SFG) rickettsioses are important neglected zoonoses throughout the world, with expanding known distribution, among which R. raoultii is an emerging member. As the clinical symptoms of R. raoultii is similar to other diseases, the accurate diagnosis of the pathogen based on the detection of genome plays a vital role in effective treatment and control at low infection levels. Due to the lack of on-site detection technology of Rickettsia raoultii in remote areas, the pathogen can only be treated in the late infection stage with specific clinical symptoms. Closed dumbbell-mediated isothermal amplification (CDA) is a method that rapidly amplifies nucleic acids under isothermal condition, which can help to resolve the problem of point of care detection for R. raoultii. The established R. raoultii detection approach based on CDA was feasible for on-site detection with merits of easy-operation, rapidity, high sensitivity, and specificity. At last, compared with PCR and LAMP, CDA method is simpler and more efficient to detect R. raoultii, which helps to improve the efficiency of detection and strengthen the prevention and control of this neglected but widely distributed disease. [ABSTRACT FROM AUTHOR]

Background: The genus Rickettsia contains the lineages spotted fever group (SFG), typhus group (TG), and transitional group (TRG). The spotted fever group Rickettsia (SFGR) is transmitted by ticks. The tick species Dermacentor nuttalli is considered the main vector carrying SFGR in Inner Mongolia. Studying the genetic diversity and population structure of Rickettsia is essential for developing effective control strategies and predicting evolutionary trends of Rickettsia. Methods: In 2019 we collected 408 D. nuttalli in the Inner Mongolia Autonomous Region, detected the percentage of Rickettsia-positive specimens, and characterized the haplotypes. From the Rickettsia-positive ticks, the gltA and ompA genes were extracted, amplified, and sequenced. Results: Ten haplotypes of the gltA gene and 22 haplotypes of the ompA gene were obtained. The phylogenetic analysis showed that the haplotypes G1–G7 and G9 of the gltA gene cluster with Rickettsia raoultii, while G8 and G10 cluster with Rickettsia sibirica. Haplotypes O1–O15, O18 and O20–O22 of the ompA gene cluster with R. raoultii, while O16 and O19 cluster with R. sibirica. The average haplotype diversity was 0.3 for gltA and 0.7 for ompA. The average nucleotide diversity was greater than 0.05. Neutrality tests were nonsignificant for Tajima's D results and Fu's Fs results. The fixation index values (FST) showed that the degree of genetic differentiation between most sampled populations was small (FST < 0.05), whereas some populations showed a medium (FST > 0.05) or large (FST > 0.15) degree of differentiation. Analysis of molecular variance (AMOVA) revealed that the variation within populations was greater than that between populations. The mismatch analysis of Rickettsia showed double peaks. Conclusions: We found two Rickettsia spp. (R. raoultii and R. sibirica). The high genetic disparity of Rickettsia allows for easy adaption to different environments. Genetic differentiation between populations is small, and Rickettsia populations do not show a geographically differentiated structure. The high rates of retention and infection of Rickettsia in D. nuttalli together with the animal husbandry exchange in Inner Mongolia gradually led to the harmonization of genetic characteristics of Rickettsia across various regions. Overall, the significant genetic diversity and geographical structure of Rickettsia in D. nuttalli are critical for SFGR control. [ABSTRACT FROM AUTHOR]

Background: The genus Rickettsia contains the lineages spotted fever group (SFG), typhus group (TG), and transitional group (TRG). The spotted fever group Rickettsia (SFGR) is transmitted by ticks. The tick species Dermacentor nuttalli is considered the main vector carrying SFGR in Inner Mongolia. Studying the genetic diversity and population structure of Rickettsia is essential for developing effective control strategies and predicting evolutionary trends of Rickettsia. Methods: In 2019 we collected 408 D. nuttalli in the Inner Mongolia Autonomous Region, detected the percentage of Rickettsia-positive specimens, and characterized the haplotypes. From the Rickettsia-positive ticks, the gltA and ompA genes were extracted, amplified, and sequenced. Results: Ten haplotypes of the gltA gene and 22 haplotypes of the ompA gene were obtained. The phylogenetic analysis showed that the haplotypes G1–G7 and G9 of the gltA gene cluster with Rickettsia raoultii, while G8 and G10 cluster with Rickettsia sibirica. Haplotypes O1–O15, O18 and O20–O22 of the ompA gene cluster with R. raoultii, while O16 and O19 cluster with R. sibirica. The average haplotype diversity was 0.3 for gltA and 0.7 for ompA. The average nucleotide diversity was greater than 0.05. Neutrality tests were nonsignificant for Tajima's D results and Fu's Fs results. The fixation index values (FST) showed that the degree of genetic differentiation between most sampled populations was small (FST < 0.05), whereas some populations showed a medium (FST > 0.05) or large (FST > 0.15) degree of differentiation. Analysis of molecular variance (AMOVA) revealed that the variation within populations was greater than that between populations. The mismatch analysis of Rickettsia showed double peaks. Conclusions: We found two Rickettsia spp. (R. raoultii and R. sibirica). The high genetic disparity of Rickettsia allows for easy adaption to different environments. Genetic differentiation between populations is small, and Rickettsia populations do not show a geographically differentiated structure. The high rates of retention and infection of Rickettsia in D. nuttalli together with the animal husbandry exchange in Inner Mongolia gradually led to the harmonization of genetic characteristics of Rickettsia across various regions. Overall, the significant genetic diversity and geographical structure of Rickettsia in D. nuttalli are critical for SFGR control. [ABSTRACT FROM AUTHOR]

Background: Ticks (Arthropoda, Ixodida), after mosquitoes, are the second most prevalent vector of infectious diseases. They are responsible for spreading a multitude of pathogens and threatening the health and welfare of animals and human beings. However, given the history of tick-borne pathogen infections in the Inner Mongolia Autonomous Region of China, surprisingly, neither the genetic diversity nor the spatial distribution of haplotypes within ticks has been studied. Methods: We characterized the haplotype distribution of Dermacentor nuttalli in four main pastoral areas of the Inner Mongolia Autonomous Region, by sampling 109 individuals (recovered from sheep) in April–August 2019. The 16S rRNA gene, cytochrome c oxidase subunit I (COI), and the internal transcribed spacer 2 region (ITS2) were amplified and sequenced from extracted DNA. Results: Twenty-six haplotypes were identified using 16S rRNA sequences, 57 haplotypes were identified with COI sequences, and 75 haplotypes were identified with ITS2 sequences. Among the three genes, total haplotype diversity was greater than 0.7, while total nucleotide diversity was greater than 0.06. Neutrality tests revealed a significantly negative Tajima's D result, while Fu's Fs was not significantly positive. Fixation index values (FST) indicated that the degree of genetic differentiation among some sampled populations was small, while for others it was moderate. Analysis of molecular variance (AMOVA) revealed that the variation within populations was greater than that among populations. The mismatch analysis of D. nuttalli exhibited double peaks. Conclusion: The genetic diversity of D. nuttalli populations in our region can likely adapt to different geographical environments, thereby leading to genetic diversity, and creating genetic differentiation among different populations. However, genetic differentiation is cryptic and does not form a pedigree geographical structure. [ABSTRACT FROM AUTHOR]

High pressure processing (HPP), as nonthermal processing technology, has the potential to increase the drying rate due to its improvement of heat and mass exchange in different processes. In this study, the moisture migration in shrimps during HPP‐vacuum‐freeze drying (HPP‐VFD) processes has been monitored by using low‐field nuclear magnetic resonance and magnetic resonance image (MRI) in comparison with hot air‐drying and VFD. Based on the T2 relaxation spectra, three water fractions corresponding to bound water (hydrogen‐bonded water), immobile water (water trapped by organization structure or cell member), and free water were observed. For group B, with increasing drying time (4 to 22 hr), the transverse relaxation times of T21, T22, and T23 were significantly decreased (76.79%, 57.78%, and 40.9%) (P < 0.05). The content of immobile water (A22) and free water (A23) decreased (81.55% and 89.07%), whereas the bound water (A21) increased (7.26%). In comparison with group B, the T21, T22, and T23 of group C showed greater decrease (83.12%, 87.12%, and 89.57% for group C) so that HPP pretreatment could shorten the relaxation time. MRI analysis further proved that HPP‐VFD drying has improved drying efficiency, and moisture migration was from the exterior to the interior part with increasing drying time. SEM analysis demonstrated that no significant damage of muscle fibers with narrower gaps was observed for groups B and C. Overall, HPP, as a pretreatment technology, could accelerate the moisture migration and improve the drying efficiency of VFD process for shrimp. Practical Application: High pressure processing (HPP) is now well known as a nonthermal processing technology and becoming increasingly acknowledged. However, there is limited information about its application in shrimp‐drying process and the moisture dynamic of shrimp subjected to high pressure processing‐assisted vacuum‐freeze drying. This study could provide valuable information regarding the moisture status and migration in HPP‐VFD shrimp monitored by LF‐NMR and MRI methods. The results showed that HPP processing at 550 MPa for 10 min can be used as an interesting method for drying pretreatment, increasing its drying rate and consequently reducing its process time, and it demonstrated that the methods used in this study had good correlation coefficient with physicochemical properties of shrimp, which may be real‐time and nondestructive monitoring methods for shrimp‐drying process. [ABSTRACT FROM AUTHOR]

Abstract: The effects of high hydrostatic pressure (HHP) treatments (200, 300, and 400 MPa for 1, 3, 5 and 10 min) on the shelling efficacy (the rate of shelling, the rate of integrity and yield of razor clam meat) and the physicochemical (drip loss, water‐holding capacity, pH, conductivity, lipid oxidation, Ca2+‐ATPase activity, myofibrillar protein content), microbiological (total viable counts) and microstructural properties of fresh razor clam (Sinonovacula constricta) were investigated. HHP treatments significantly (P < 0.05) increased shelling efficiency, water‐holding capacity, pH, conductivity, and lipid oxidation, and HHP‐treated razor clam showed lower levels of microorganisms and drip loss than untreated razor clam. Levels of thiobarbituric acid reacting substances (TBA) in HHP‐treated razor clam were greatly increased (up to 0.93 ± 0.09 mg MDA/kg at 400 MPa for 10 min) which was caused by the formation of hydroperoxides during HHP treatment. All HHP treatments were found to have adverse effects on the activity of Ca2+‐ATPase and the content of myofibrillar protein (MP), which might be due to the substantial damage to the tertiary structure of proteins at high pressure. Moreover, scanning electron microscopy (SEM) revealed the compaction of the muscle fibers and a decrease in the extracellular space with increasing pressure and holding time. This phenomenon was mainly correlated with the compaction of muscle fibers and denaturation, aggregation, and gelation of muscle protein triggered by high pressure. In general, HHP could be applied as a safe and effective nonthermal technology to produce high‐quality shelled razor clam. Practical application: High hydrostatic pressure (HHP) is now well known as a nonthermal processing technology and becoming increasingly acknowledged. However, it has not been widely applied to shell seafood due to its uncertain influence on its quality and shelling property. This study could provide valuable information regarding the shelling efficacy, physicochemical properties, and microstructure of razor clam treated by HHP. And it demonstrated that HHP showed a positive impact on quality of razor clam treated by HHP. [ABSTRACT FROM AUTHOR]

The article presents a study which synthesized and characterized the amphiphilic random copolymer composed of monomeric units of poly (butyl acrylate) and poly (maleic acid). It examined the emulsion polymerization kinetics of styrene stabilized by the copolymer. It mentioned the use of dynamic light scattering technique and transmission electron microscope to measure the sizes and shapes of the particles.