Here, we describe for the first time the prevalence and genetic properties of Bartonella organisms in wild rodents in Japan. Recently, genomics, transcriptomics, and proteomics studies were used in this traditional field and induced much progress. Phylogenetic relationships within the Bartonella genus based on the groEL gene. Bartonella bacilliformis was used as an outgroup. They are Gram-negative bacteria and are characterized by motile structures known as flagella.. According to the genetic identity values proposed by La Scola et al. 2013 May-June, Katie L. Burke, “In the News". Samples negative for the IRBP gene by PCR were subsequently submitted to another internal control PCR targeting the GAPDH gene (29), using GAPDH-F (5′-CCTTCATTGACCTCAACTACAT-3′) and GAPDH-R (5′-CCAAAGTTGTCATGGATGACC-3′) primers and the same concentration of reagents described for IRBP PCR (except DMSO). Humans generally become infected by coming in contact with contaminated soil or water, often while In addition, the Bartonella species nucleotide diversity revealed in our study was higher than that reported in B. grahamii detected in rodents belonging to the species Myodes rutilus, Microtus fortis, and Apodemus agrarius (π = 0.012; Hd = 0.700) sampled in 4 sites from China (58). detected in the infected rodents, (iii) identify and characterize the Bartonella spp. Rodents were trapped using cage and breakâback traps and purchased from local markets in June and December 2006 in four Lao provinces. The tree was inferred by using the maximum likelihood (ML) method with the GTR+G+I model. Euthanasia of animals was performed for taxonomic identification and/or diagnosis of parasites. Diagnostic Microbiology Micrococci are catalase-positive, oxidase-positive, strictly aerobic gram-positive cocci that grow in clusters. Herein we reported the occurrence of Bartonella spp. Additionally, we performed a concatenated phylogenetic analysis with the sequences which were simultaneously positive in PCR assays based on gltA, ftsZ, and groEL genes. Polymorphism and genetic diversity of Bartonella spp. Hantaviruses Causing HPS Sin Nombre virus (SNV) was first isolated from rodents collected on the premises of one of the initial HPS patients in the Four Corners region. (2019) Leishmaniasis In: Hof H, Schlüter D, Dörries R, eds Duale Reihe Medizinische Mikrobiologie. Some studies have reported a high prevalence of infection in rodent communities, which can reach up to 82% (15). MA, Mata Atlântica biome; CE, Cerrado biome; CA, Caatinga biome. Rodents that have undergone an adventitious viral infection should not be used for passaging cell lines or as sources of tissues and fluids for subsequent experiments. Rodents were chemically immobilized using a combination of ketamine hydrochloride (100 mg/ml) and acepromazine (10 mg/ml) intramuscularly. The sequences detected in the present study are highlighted in bold. 7th, completely revised and expanded edition.
Backgrounds. when they analyzed rodents from the state of Mato Grosso do Sul, Brazilian Pantanal (21). Copyright © 2020 LoveToKnow. The current study aimed to assess the protozoan infection of rodents in Boyer-Ahmad district, southwestern Iran.
Materials and Methods. PCR amplifications were conducted in low-profile multiplate unskirted PCR plates (Bio-Rad, CA, USA) using a CFX96 thermal cycler (Bio-Rad, CA, USA). Bartonella genetic diversity and distribution analysis.The alignment sequences of the gltA, ftsZ, and groEL genes amplified in the present study were utilized to calculate the nucleotide diversity (π), the polymorphic level (haplotype diversity [Hd]), the number of variable sites (v), and the average number of nucleotide differences (K) using DnaSP (v. 5.10) (using default parameters) (42). MA, Mata Atlântica biome; CE, Cerrado biome. Additionally, the logistic regression analysis performed by André et al. Therefore, more studies are needed to assess the genetic diversity of Bartonella spp. Bartonella bacilliformis was used as an outgroup. Strains are used in laboratory experiments. Rodents, for example, are a branch of mammals that split off after the end of the period when the clade Dinosauria stopped being the dominant terrestrial vertebrates 66 million years ago. Spirilla Bacteria Definition, Examples, Shape, Diseases What is Spirilla (Singular, Spirillum)? Evaluation of DNA extraction quality.In order to evaluate the quality of DNA samples, each sample of spleen-extracted DNA was used as a template in an internal control PCR targeting the mammalian IRBP (interphotoreceptor retinoid binding protein) gene (28). Definition of microorganisms: Organisms that cannot be seen with the naked eye and without microscopes are called microorganisms. in Brazil is caused by B. henselae and B. quintana (22, 23). The efficiency mean of qPCR assays was E = 96.6% (90.8% to 101%; SD, ±2.78), slope = −3.407, and r2 = 0.994. Only the haplotype numbers 1 (with four sequences), 5 (with two sequences), and 14 (with two sequences) had one or more sequences (Fig. arupensis in wild rodents (21). Several studies suggest that Bartonella spp. arupensis (9), B. washoensis (10), B. rochalimae (11), and B. doshiae (12) have been implicated in human illnesses. Among 1,341 blood samples from rodents that were trapped in Southeast Asia between 2008 and 2010, we found a prevalence of Bartonella infection ranging from 9.6 to 11.9%.Bartonella species identified (143 isolates) included B. elizabethae, B. coopersplainsensis, B. phoceensis, B. queenslandensis, B. rattimassiliensis, B. tribocorum, and three new putative Bartonella species. However, it was lower than that demonstrated in Myodes glareolus (π = 0.077) sampled in Paris, France (49). The phylogenetic analysis of partial sequences of gltA, ftsZ, and groEL genes when analyzed separately or by concatenated tree, showed a clear separation between the sequences obtained in the present study and other Bartonella spp. This microscopic parasite causes a diarrheal illness in the small intestines of vertebrates called giardiasis. Except for the sequence 2741 (belonging to haplotype 1) detected in Thrichomys laurentius sampled in the Caatinga biome, all of the haplotypes identified were unique to a particular biome. These findings reveal a higher sensitivity of qPCR than of cPCR assays, highlighting the use of multiple approaches in order to increase the sensitivity of Bartonella detection. Except for gltA sequences, all amplified sequences (ftsZ and groEL genes) showed a low percentage of identity (ranging from 91 to 93%) with other Bartonella spp., mainly with the B. vinsonii species complex. Only sequences with expected sizes and without background in the electropherograms were submitted to BLASTn, phylogenetic, and diversity analyses. Because of this, a One Health approach (60) is required to elucidate the vector ecology and transmission dynamics of these bacteria detected among wild Brazilian rodents and the possible role in human infections. within rodent communities in Brazil. Among the 20 gltA sequences analyzed (distributed in 11 different sites), 15 different haplotypes were identified showing π = 0.02459, Hd = 0.958, and K = 8.1894. The Akaike information criterion (AIC) available in MEGA 5.05 (41) was applied to identify the most appropriate model of nucleotide substitution. have adapted to rodent species. They are mainly found on soil, food, or contaminated water. Because of the high diversity and ecological plasticity displayed by this bacterial group, new approaches have been proposed to help in solving some conflicts between the definitions of new species (3, 4). in rodents is much more frequent and widespread that previously recognized, and this trait may be also influenced by the characteristics of rodent fauna and environment from different Brazilian biomes. However, rodents have become the animal model of choice for studies of immunocompetence, and most of our knowledge of the development of the immune system has come from studies using mice or rats. in qPCR. In conclusion, our study showed that the occurrence of Bartonella bacteria in rodents is much more frequent and widespread than previously recognized. The genus Yersinia includes Yersinia pestis, the cause of plague; Yersinia enterocolitica, an important cause of human diarrheal disease; and several others considered nonpathogenic for humans.Pasteurella are primarily animal pathogens but Pasteurella ⦠Although the Kosoy et al. Among the 117 (25.6%) samples positive for Bartonella spp. Interestingly, we found differences in the occurrence of Bartonella spp. received a scholarship from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior. However, limited information about the epidemiological, genetic diversity, distribution, and ecological aspects of Bartonella infections in Brazil is available as yet. were detected in 25.6% (117/457) of rodent spleen samples analyzed, and this occurrence varied among different biomes. Median-joining network of gltA haplotypes detected in wild rodents sampled in 11 different sites in Brazil. Analysis of the phylogenetic relationship of gltA sequences performed by Bayesian inference and maximum likelihood showed that the Bartonella species detected in rodents from Brazil was closely related to the phylogenetic group A detected in other cricetid rodents from North America, probably constituting only one species. Last, in order to correctly determine the nucleotide composition, the electropherograms were also submitted to the Phred-Phrap program (35) and compared through alignment in BioEdit. comprise an ecologically successful group of microorganisms that infect erythrocytes and have adapted to different hosts, which include a wide range of mammals, besides humans. The consensus sequences were obtained through the analysis of the sense and antisense sequences using the CAP3 program (http://mobyle.pasteur.fr/cgi-bin/MobylePortal/portal.py) (34). DNA among rodents trapped in five distinct Brazilian biomes. in humans. infecting wild rodents in Brazil is similar to that reported for B. grahamii (a Bartonella species with high genetic diversity) upon comparison with gltA sequences obtained in strains from Asia (π = 0.02154; Hd = 0.943) and North America and Europe (π = 0.01427; Hd = 0.889) (58). However, such findings should be analyzed with prudence, since in the present study, the hosts were not deeply sampled to accurately determine Bartonella abundance in each biome. The definition of rodent is something gnawing, or something related to a small mammal with constantly growing incisor teeth used for gnawing. Spleen tissues from 500 rodents were collected and stored in DNase- and RNase-free microtubes containing ethanol and maintained at −20°C until DNA extraction. live organisms) are ⦠A trap intended to capture a mouse or a rat is an example of a rodent trap. The phylogenetic analyses performed using different methods (BI and ML) yielded similar tree topologies and the same relationships for all genes analyzed in this study. were known; however, in the last few decades, the number of species in this genus has been significantly expanded. The Bayesian tree was chosen to represent the phylogenetic relationships among the gltA sequences (Fig. The immunological characteristics of these Taiwan isolates were compared with those of other ⦠5). Consequently, a given genotype may circulate in different rodent species coexisting in a certain biome, as described for hemoplasmas, another group of pathogens also transmitted between rodents by arthropods (55). These studies reported the first isolation of B. queenslandensis (5/26) and B. tribocorum (1/26) in synanthropic rodents (20) and the first detection of Bartonella vinsonii subsp. Spirilla (singular, Spirillum) are a group of bacteria characterized by a corkscrew (spiral) appearance. Accuracy assessment, Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice, BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT, MrBayes 3: Bayesian phylogenetic inference under mixed models, A rapid bootstrap algorithm for the RAxML Web servers, Creating the CIPRES science gateway for inference of large phylogenetic trees, p 1-8, Proceedings of the Gateway Computing Environments Workshop (GCE), MEGA 5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods, DnaSP v5: a software for comprehensive analysis of DNA polymorphism, Median-joining networks for inferring intraspecific phylogenies, POPART: full-feature software for haplotype network construction, Distribution, diversity, and host specificity of. Wild rodents are reservoirs of various zoonotic diseases, such as toxoplasmosis, babesiosis, and leishmaniasis. Additionally, the Bartonella genotypes detected were closely related to those detected in other Cricetidae rodents from Americas, probably representing an unique species. Supplemental material for this article may be found at http://dx.doi.org/10.1128/AEM.02447-16. Last, the nucleotide diversity observed in the ftsZ sequences (π = 0.037; Hd = 1.000) amplified in the present study was higher than those previously reported within ftsZ sequences of B. grahamii amplified in France (π = 0.016) (49) and China (π = 0.010; Hd = 1.000) (58). Quality of DNA samples and qPCR assay.Out of the 500 rodent spleen samples analyzed, 457 were positive for the internal control (IRBP gene) PCR (Table 1). These findings revealed that Bartonella detected in the present study are a monophyletic and widespread group among seven different rodent species trapped in three distinct Brazilian biomes. Trends in Microbiology 11:318â321. detected in wild rodents (Sigmodon hispidus and Oryzomys palustris) and fleas (Polygenis gwyni) from the United States and previously named phylogenetic group A (45) (92.6% identity with B. vinsonii), a frequently found but not very well molecularly characterized Bartonella genogroup isolated from S. hispidus in the southeastern United States. Purified amplified DNA fragments from positive samples were submitted to sequence confirmation in an automatic sequencer (ABI Prism 310 genetic analyzer; Applied Biosystems/PerkinElmer) in both directions. MA: Mata Atlântica; CE: Cerrado biomes. Among them, only 25 samples were simultaneously positive in gltA, ftsZ, and groEL cPCR assays. Additionally, the concatenated phylogenetic tree (gltA, ftsZ, and groEL genes) generated by the maximum likelihood method using B. vinsonii and B. elizabethae species complexes, B. grahamii, B. taylorii and B. tamiae species, and three sequences amplified in our study showed that the Bartonella detected in rodents trapped in distinct Brazilian biomes are more closely related to the B. vinsonii species complex (Fig. associated with rodents coupled with the fact that Rodentia are widely distributed in different habitats and represent the largest order of mammals (24) emphasizes the need for further studies to assess the pathogenicity of rodent-associated Bartonella spp. was detected in different rodent species derived from distinct (and distant) Brazilian biomes, showing a probable dominance of this genotype. In contrast to the traditional focus on avoiding or reducing negative welfare states, modern animal welfare concepts highlight the importance of promoting positive welfare states in laboratory animals. The Norway rat, roof rat, and house mouse are considered commensal rodents because of their intimate relationships with humans. Many commonly named groups, rodents and insects for example, are clades because, in each case, the group consists of a common ancestor with all its descendant branches. From the Latin prefix com- (with, together, jointly) + the Latin adjective mensalis (of the table). Comparative Immunology, Microbiology and Infectious Diseases Volume 72, October 2020, 101515 Bartonella spp. The maximum likelihood (ML) phylogenetic analysis was inferred with RAxML-HPC BlackBox (v. 7.6.3) (the bootstrap values higher than 50% were accessed with 1,000 replicates) (39); both analyses were performed through the CIPRES Science Gateway (40). Serial dilutions were performed with the aim of constructing standard curves with different concentrations of plasmid DNA (pIDTSmart; Integrated DNA Technologies) (2.0 × 107 to 2.0 × 100 copies/μl), which encoded a 83-bp Bartonella henselae nuoG gene fragment (30). Bartonella infection in rodents usually results in a persistent and subclinical bacteremia lasting for months, characterizing the infected rodents as important sources of infection to other susceptible hosts, including humans (13, 14). Any of various mammals of the order Rodentia, such as a mouse, rat, squirrel, or beaver, characterized by large incisors used for gnawing or nibbling. Definition of a Parasite Parasites are plants or animals that live on or in a host getting their nutrients from that host. Any of a very large order (Rodentia) of gnawing mammals, including rats, mice, squirrels, and beavers, characterized by constantly growing incisors adapted for gnawing or nibbling; esp., in popular usage, a rat or mouse. Nevertheless, the presence of bartonellae infection Any of various very numerous, mostly small mammals of the order Rodentia, having large front teeth used for gnawing. All Rights Reserved. An animal’s capacity to suffer is a prerequisite for any animal welfare concern, and the minimization of suffering is a key aim of refinement research. Bartonella henselae DNA obtained from a naturally infected cat was used as a positive control (30). Another definition is: âthe process by which biological agents (i.e. salinicola is a much smaller rodent from the salt-lagunas of Argentina. Leptospira spp. Rodents (from Latin rodere, "to gnaw") are mammals of the order Rodentia (/roÊËdÉnÊÉ/), which are characterized by a single pair of continuously growing incisors in each of the upper and lower jaws. The sequences of the gltA gene were submitted to a median-joining network (43) inferred using the Population Analysis with Reticulate Trees (popART) (v. 1.7) (using default parameters) (44). by qPCR. (phylogenetic group A and strain R-phy1) detected in rodents belong to the Cricetidae family and fleas parasitizing some of these rodents in North America (1, 45, 53). On the other hand, a lower occurrence of Bartonella was observed in rodents caught in Amazon forest (3.2% [1/31]). Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback, Print ISSN: 0099-2240; Online ISSN: 1098-5336, Public and Environmental Health Microbiology, Sign In to Email Alerts with your Email Address. associated with rodents in human health should be assessed in future studies. All samples negative for the IRBP gene PCR were also negative for the GAPDH gene PCR and were excluded from subsequent analyses. by qPCR, only 25 (21.3%) were simultaneously positive by gltA, ftsZ, and groEL cPCR assays. Rodents A mouse or a rat strain is a group of animals that is genetically uniform. Rodent is defined as a small mammal that has a pair of incisor teeth that are constantly growing and that are used for gnawing. Although the genetic diversity of this group of pathogens infecting rodents has been very well documented (49–52), this high variability has caused a laborious challenge in the taxonomic constitution of this group when the old criteria, such as DNA-DNA hybridization, comparison of 16S rRNA gene sequences, and phenotypical characteristics, are used to describe Bartonella spp. In our study, we performed molecular and phylogenetic analyses of Definition, importance and microbiological characteristics of laboratory animals. previously described in GenBank, corroborating the low percentage of identity obtained by BLASTn analysis. The other phylogenetic trees are available in Fig. Topic 3. IMPORTANCE In the present study, we reported the occurrence of Bartonella spp. Bartonella spp. The sequences detected in the present study are highlighted in bold. The haplotype diversity results of the additional genes are shown in Table 3. Rodents are reservoirs of about two-thirds of Bartonella spp. To determine the limit of detection from the qPCR assay, the standard curves generated by 10-fold dilutions were used to determine the amount of DNA that could be detected with 95% sensitivity (31). Microorganisms are everywhere. Copyright © 2016, American Society for Microbiology. Rodents are reservoirs of about two-thirds of Bartonella spp. Microorganisms are found in water in the air, in animals on We are thankful to Gener Tadeu Pereira (FCAV/UNESP) and to Aline Meira Bonfim Mantellatto (FCAV/UNESP) for assistance in statistical and diversity and distribution analyses, respectively. All specimens were photographed, vouchered (specimens will be registered in the Australian National Wildlife Collection, CSIRO Sustainable Ecosystems, Canberra, Australia) and identified subsequently (by KA) [ 4 ]. Occurrence and absolute quantification of Bartonella spp. research group (45, 48) showed host specificity for the phylogenetic group A, our results demonstrate that Bartonella spp. The sequences detected in the present study are highlighted in bold. Phylogenetic relationships within the Bartonella genus based on the ftsZ gene. that have been reported worldwide. 4), compared with other previously described Bartonella sequences, showed a clear separation in both genes analyzed, constituting a monophyletic group, clustering closely to the Bartonella vinsonii complex. These findings reveal that the genetic diversity of Bartonella detected in wild rodents from Brazil is similar to or higher than that in some strains of B. grahamii detected around the world. DNA extraction.DNA was extracted from 10 mg of each rodent spleen tissue using the DNeasy blood and tissue kit (Qiagen, Valencia, CA, USA), according to the manufacturer's instructions. We also noted subtle intergene variation in levels of diversity among the ftsZ (π = 0.037) and groEL (π = 0.027) genes compared to that for gltA. The same Bartonella sp. We captured 685 wild rodents throughout Japan (in 12 prefectures) and successfully isolated Bartonella organisms from 176 of the 685 rodents (isolation rate, 25.7%). Eight-one (69.2%), 56 (47.8%), and 35 (29.9%) samples were positive in ftsZ, gltA, and groEL cPCR assays, respectively. The diversity analysis performed in the present study suggests that the genetic diversity detected in gltA sequences of Bartonella spp. in wild rodents from Caatinga, Cerrado, Atlantic forest, and Amazon forest biomes in Brazil. All Rights Reserved. An intimate relationship. Phylogenetic relationships within the Bartonella genus based on a concatenated tree (gltA, ftsZ, and groEL genes). Last, the Bartonella species genogroup identified in the present study formed a monophyletic group that included Bartonella samples from seven different rodent species distributed in three distinct biomes.
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