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Background Four species within Anaplasma genus are emerging zoonotic pathogens, which are transmitted by ticks and generate veterinary and public health concerns. Here, we performed a molecular survey of Anaplasma in Ankang, Northwest China. Methods Hard ticks were collected and identified using morphological and molecular methods. Human-pathogenic Anaplasma species were tested using nested polymerase chain reaction. The nearly complete rrs , gltA , and groEL genes sequences from revealed Anaplasma species were amplified and sequenced to determine their molecular characteristics and their phylogeny. Results All ticks collected in Ankang belonged to the Rhipicephalus microplus . Novel unclassified Anaplasma strains genetically related to A. platys and A. capra were detected in these ticks. Co-infection of these two organisms was also found. The novel unclassified Anaplasma strains identified in this study formed a distinct phylogenetic lineage based on the groEL gene and two lineages based on the gltA gene within A. platys and related strains group. The revealed A. capra strains identified in this study were most closely related to those detected in humans and other vertebrate animals. Conclusion We revealed the presence of A. capra , a novel human pathogens in R. microplus ticks in previously unrecognized endemic regions. We also detected a novel unclassified Anaplasma species genetically related to A. platys . The epidemiology of anaplasmosis caused by these two Anaplasma species in humans should be assessed in future studies.

Background Clustered regularly interspaced short palindromic repeats interference (CRISPRi) has provided an efficient approach for targeted gene inhibition. A non-model microorganism Halomonas species TD01 has been developed as a promising industrial producer of polyhydroxyalkanoates (PHA), a family of biodegradable polyesters accumulated by bacteria as a carbon and energy reserve compound. A controllable gene repression system, such as CRISPRi, is needed for Halomonas sp. TD01 to regulate its gene expression levels. Results For the first time CRISPRi was successfully used in Halomonas sp. TD01 to repress expression of ftsZ gene encoding bacterial fission ring formation protein, leading to an elongated cell morphology with typical filamentous shape similar to phenomenon observed with Escherichia coli . CRISPRi was employed to regulate expressions of prpC gene encoding 2-methylcitrate synthase for regulating 3-hydroxyvalerate monomer ratio in PHBV copolymers of 3-hydroxybutyrate (HB) and 3-hydroxyvalerate (HV). Percentages of HV in PHBV copolymers were controllable ranging from less than 1 to 13%. Furthermore, repressions on gltA gene encoding citrate synthase channeled more acetyl-CoA from the tricarboxylic acid (TCA) cycle to poly(3-hydroxybutyrate) (PHB) synthesis. The PHB accumulation by Halomonas sp. TD01 with its gltA gene repressed in various intensities via CRISPRi was increased by approximately 8% compared with the wild type control containing the CRISPRi vector without target. Conclusions It has now been confirmed that the CRISPRi system can be applied to Halomonas sp. TD01, a promising industrial strain for production of various PHA and chemicals under open and continuous fermentation process conditions. In details, the CRISPRi system was successfully designed in this study to target genes of ftsZ , prpC and gltA, achieving longer cell sizes, channeling more substrates to PHBV and PHB synthesis, respectively. CRISPRi can be expected to use for more metabolic engineering applications in non-model organisms.

Ixodes ricinus and Dermacentor reticulatus ticks are important reservoirs and vectors of pathogens. The aim of the present study was to investigate the dynamic of the prevalence and genetic diversity of microorganisms detected in these tick species collected from two ecologically diverse biotopes undergoing disparate long-term climate condition. High-throughput real time PCR confirmed high prevalence of microorganisms detected in sympatrically occurring ticks species. D. reticulatus specimens were the most often infected with Francisella -like endosymbiont (FLE) (up to 100.0%) and Rickettsia spp. (up to 91.7%), while in case of I. ricinus the prevalence of Borreliaceae spirochetes reached up to 25.0%. Moreover, pathogens belonging to genera of Bartonella , Anaplasma , Ehrlichia and Babesia were detected in both tick species regardless the biotope. On the other hand, Neoehrlichia mikurensis was conformed only in I. ricinus in the forest biotope, while genetic material of Theileria spp. was found only in D. reticulatus collected from the meadow. Our study confirmed significant impact of biotope type on prevalence of representatives of Borreliaceae and Rickettsiaceae families. The most common co-infection detected in D. reticulatus was Rickettsia spp. + FLE, while Borreliaceae + R. helvetica was the most common in I. ricinus . Additionally, we found significant genetic diversity of R. raoultii gltA gene across studied years, however such relationship was not observed in ticks from studied biotopes. Our results suggest that ecological type of biotope undergoing disparate long-term climate conditions have an impact on prevalence of tick-borne pathogens in adult D. reticulatus and I. ricinus .

Ticks are the second most significant hematophagous ectoparasites, serving as vectors for numerous infectious diseases worldwide, surpassed only by mosquitoes. This study aimed to molecularly characterize ticks infesting cows and their associated pathogens in Khyber Pakhtunkhwa, Pakistan, with a focus on assessing the potential risk of zoonotic disease transmission. Between January 2023 and June 2024, 240 cows from six districts: Buner, Swat, Lower Dir, Upper Dir, Shangla, and Chitral, were examined for tick infestations. DNA was extracted using the phenol-chloroform method, and pathogen detection was performed via PCR targeting the gltA gene, while tick species were confirmed by amplifying partial 12 S rDNA sequences. The study revealed an average tick intensity of 12.73 ticks per infested cow and a mean abundance of 10.45 ticks per examined cow. In total, 2,507 ticks were collected and morphologically identified as Rhipicephalus microplus, comprising 915 females, 801 males, 510 nymphs, and 281 larvae. The highest tick burdens were observed in Buner and Swat, followed by Lower Dir, Upper Dir, and Shangla, with Chitral showing the lowest prevalence. Molecular screening detected multiple pathogens, notably the human pathogen Anaplasma capra and the cow pathogen Anaplasma marginale ; co-infections were occasionally observed. This paper presents the first phenotypic and phylogenetic characterization of A. capra in ticks from cows in Pakistan, emphasizing the zoonotic transmission risk. These findings advance our understanding of R. microplus associated pathogen risks and underscore the need for enhanced surveillance in the region.

Sheep and goats constitute a critical component of the Egyptian agricultural economy, serving as primary sources of milk, meat, and wool for rural populations. This study was conducted to detect and characterize infections caused by Borrelia theileri , Rickettsia aeschlimannii , Mycoplasma ovis , and Mycoplasma wenyonii in small ruminants from southern Egypt using molecular methods. The identification of these pathogens underscores potential threats to livestock productivity and raises public health concerns, particularly due to the zoonotic nature of Rickettsia aeschlimannii . The detection of these pathogens was performed using polymerase chain reaction assays targeting the flaB, gltA , and 16S rRNA genes, enabling precise identification and genetic characterization of the infectious agents. Confirmation and genotyping were achieved through bidirectional sequencing and phylogenetic analysis. Blood samples were collected from 300 sheep and 300 goats across two governorates in southern Egypt. The overall infection rate of Borrelia theileri was 2.34% in sheep and 1.00% in goats. For Rickettsia aeschlimannii , the infection rates were 2.00% in sheep and 1.33% in goats. Mycoplasma ovis was detected in 1.33% of sheep and 1.00% of goats, while Mycoplasma wenyonii was present in 1.33% of sheep but not detected in goats. Genetic analysis indicated that the Rickettsia aeschlimannii gltA gene and the Borrelia theileri flaB genes were identical to those found in dogs, camels, and ticks in southern Egypt. The Mycoplasma wenyonii 16S rRNA gene matched cattle samples from the same region, and the Mycoplasma ovis 16S rRNA gene was identical to the one found in camels. Beyond confirming the presence of tick-borne diseases in small ruminants, these results show that tick-borne diseases are genetically related to other hosts present in the area. These findings highlight the need for integrated vector surveillance and host health assessments to evaluate zoonotic risk and subclinical impacts.

is an emerging pathogen, which can infect ruminants and humans. This study was conducted to determine the occurrence of in the blood samples of sheep and goats in China. Using nested polymerase chain reaction (nested-PCR) targeting the gene and conventional PCR targeting the heat shock protein ( ) gene and the major surface protein4 gene ( ), was detected in 129 (8.9%) of 1453 sheep and goat blood samples. The positive rate was higher in goats (9.4%, 89/943) than in sheep (7.8%, 40/510) (χ = 1.04, > 0.05, = 1). For sheep, was found in 17 sites from 2 provinces. The prevalence was 28.6% in sheep from Liaoning province, which was higher than in Henan Province (7.3%). For goats, was detected in 35 sites from 7 provinces. The prevalence varied from 0 to 19.4% in the goat sites examined. The prevalence rates were 19.4, 19.3, 10, 8.8, 6.8, 1.8, and 0% in goats from Guizhou province, Henan Province, Inner Mongolia Autonomous Region, Shanxi Province, Xinjiang Uygur Autonomous Region, Yunnan province, and Gansu province, respectively. Based on the analysis of the citrate synthase gene ), two variants were identified. Variant I showed a high sequence similarity to the , which were previously reported in sheep, goats, , and humans. Variant II was only found in Luoyang, Anyang, and Sanmengxia, of Henan province. To our knowledge, this is the first detection of this variant of in sheep and goat blood in China. Phylogenetic analysis based on and genes showed that the sp. sequences clustered independently from and other species with high bootstrap values. We found DNA in sheep and goats in China, providing evidence that sheep and goats can be infected by . We also found that this zoonotic pathogen is widely distributed in China. This study provides information for assessing the public health risks for human anaplasmosis.

Bartonella species are emerging human pathogens. Bats are known to carry diverse Bartonella species, some of which are capable of infecting humans. However, as the second largest mammalian group by a number of species, the role of bats as the reservoirs of Bartonella species is not fully explored, in term of their species diversity and worldwide distribution. China, especially Northern China, harbors a number of endemic insectivorous bat species; however, to our knowledge, there are not yet studies about Bartonella in bats in China. The aim of the study was to investigate the prevalence and genetic diversity of Bartonella species in bats in Northern China. Bartonella species were detected by PCR amplification of gltA gene in 25.2% (27/107) bats in Mengyin County, Shandong Province of China, including 1/3 Rhinolophus ferrumequinum, 2/10 Rhinolophus pusillus, 9/16 Myotis fimbriatus, 1/5 Myotis ricketti, 14/58 Myotis pequinius. Phylogenetic analysis showed that Bartonella species detected in bats in this study clustered into ten groups, and some might be novel Bartonella species. An association between Bartonella species and bat species was demonstrated and co-infection with different Bartonella species in a single bat was also observed. Our findings expanded our knowledge on the genetic diversity of Bartonella in bats, and shed light on the ecology of bat-borne Bartonella species.

Tick-borne diseases have been an increasing threat to human and animal health all over the world. Anaplasmosis is one of the emerging tick-borne diseases and has zoonotic potential. A new novel species, which was detected in China in 2010–2012 and provisionally named Anaplasma capra in 2015, causes zoonotic infections and infects many different animal species. In this study, we investigated the presence of A. capra in domestic ruminants from Turkey. A total of 468 blood samples (cattle, sheep, and goat) were examined by the gltA gene-specific nested polymerase chain reaction, revealing the presence of A. capra in six samples (1.28%): one of them from cattle (0.41%) and the other five from sheep (3.22%). According to DNA sequences results of the gltA gene, A. capra isolates identified in the present study were shown high nucleotide similarity with A. capra isolates detected from different hosts. However, the nucleotide differences were detected in the same nucleotide positions between A. capra isolates. For this reason, we thought that at least two different A. capra genotypes could be circulating in the world. As a result, it is seen that A. capra, which was determined to be a new species with zoonotic potential, was revealed in European and Asian countries and in different hosts. In order to raise awareness about human anaplasmosis infections, it is important to reveal the prevalence of the species in the world. The emergence of A. capra in Turkey reveals the need for a re-evaluation of the human and animal health risk analysis in terms of anaplasmosis.

The Bartonella genus consists of neglected pathogens associated with potentially transfusional-transmitted and fatal human diseases. We aimed to evaluate Bartonella sp. prevalence in 500 blood donors and compare the results with the data already published about these samples. We used molecular diagnostic methods to detect Bartonella sp.-DNA from blood and liquid culture samples: (A) conventional PCR for two gene regions, the ITS targeting the genus Bartonella and the specific gltA Bartonella henselae ; (B) nested PCR for the ftsZ gene and (C) qualitative real-time PCR for the gltA gene, both B . henselae specific. We obtained 30/500 (6%) DNA detections from the blood samples; 77/500 (15.4%) DNA detections from liquid culture samples and five (1%) samples had DNA detection from both. In total, we detected B . henselae DNA from 102/500 (20.4%) donors. The samples used in this study had already been submitted for Bartonella sp.-DNA detection using only a conventional PCR in liquid culture. Sixteen samples (3.2%) were positive previously, and from these 16 samples, 13 were negative in the new investigation. We concluded that the use of liquid culture combined with different molecular tests increases the possibility of detecting Bartonella sp.-DNA, but the tests do not avoid false-negative results. More than a fifth of blood donors had at least one PCR that detected Bartonella sp.-DNA among the eight molecular reactions performed now (four reactions in whole blood and four in liquid culture). Seven percent had B . henselae -DNA detection for two or more distinct regions. Considering the results obtained previously, the DNA of Bartonella spp. was detected or the agent isolated in 23% of analyzed blood donors. The results establish that the low bacteremia and the fastidious characteristics of the bacterium are challenges to laboratory diagnosis and can make it difficult to confirm the infection in patients with bartonelloses.

Anaplasma and Ehrlichia are tick-borne bacterial pathogens that cause anaplasmoses and ehrlichioses in humans and animals. In this study, we examined the prevalence of Anaplasma and Ehrlichia species in ticks and domesticated animals in Suizhou County, Hubei Province in the central China. We used PCR amplification and DNA sequencing of the 16S rRNA, gro EL, and glt A genes to analyze. We collected 1900 ticks, including 1981 Haemaphysalis longicornis and 9 Rhipicephalus microplus , 159 blood samples of goats (n = 152), cattle (n = 4), and dogs (n = 3) from May to August of 2023. PCR products demonstrated that Anaplasma bovis , Anaplasma capra , and an Ehrlichia species were detected in the H. longicornis with the minimum infection rates (MIR) of 1.11%, 1.32%, and 0.05%, respectively; A. bovis , A. capra , and unnamed Anaplasma sp. were detected in goats with an infection rate of 26.31%, 1.31% and 1.97%, respectively. Anaplasma and Ehrlichia species were not detected from cattle, dogs and R. microplus ticks. The genetic differences in the gro EL gene sequences of the Anaplasma in the current study were large, whereas the 16S rRNA and glt A gene sequences were less disparate. This study shows that ticks and goats in Suizhou County, Hubei Province carry multiple Anaplasma species and an Ehrlichia species, with relatively higher infection rate of A. bovis in goats. Our study indicates that multiple Anaplasma and Ehrlichia species exist in ticks and goats in the central China with potential to cause human infection.