Distribution of rodents sampled in different Brazilian biomes. Additionally, the number of Bartonella spp. in the present study (54). This is most easily observed in microbiology where strains are derived from a single cell colony and are typically quarantined by the physical constraints of a Petri dish. in wild rodents.Among all DNA rodent spleen samples analyzed, 25.6% (117/457) were positive for Bartonella spp. The identification and understanding of the distribution of this important group of bacteria may allow the Brazilian authorities to recognize potential regions with the risk of transmission of these pathogens among wild and domestic animals and humans. were known; however, in the last few decades, the number of species in this genus has been significantly expanded. In a nutshell, our study showed that the occurrence of Bartonella spp. Trends in Microbiology 11:318–321. Bartonella spp. Definition, importance and microbiological characteristics of laboratory animals. Bartonella bacilliformis was used as an outgroup. The tree was inferred by using the maximum likelihood (ML) method with the GTR+G+I model. detected in rodents trapped in America. Screening and quantification of Bartonella species DNA (qPCR assay).A previously described broad-range qPCR protocol based on the nuoG gene was used with the aim of detecting and quantifying Bartonella species DNA copies (number of copies per microliter) (30). Median-joining network of gltA haplotypes detected in wild rodents sampled in 11 different sites in Brazil. described to date; and some of them have been implicated as causative agents of human diseases. infecting wild and synanthropic rodents using gltA, ftsZ, and groEL genes, and (iv) analyze the haplotype diversity of Bartonella sequences detected. 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). The Bayesian tree was chosen to represent the phylogenetic relationships among the gltA sequences (Fig. Distribution and rodent species sampled.Between 2000 and 2011, different rodent species (n = 52) were trapped in five Brazilian biomes (Fig. L.R.G. Rodents (from Latin rodere, "to gnaw") are mammals of the order Rodentia (/roʊˈdɛnʃə/), which are characterized by a single pair of continuously growing incisors in each of the upper and lower jaws. Additionally, the Bartonella genotypes detected were closely related to those detected in other Cricetidae rodents from Americas, probably representing an unique species. However, gltA sequences detected in infected rodents were closely related to Bartonella spp. described, the criteria used for separation of new genotypes of previously described species remain a big issue (3, 4). Recent studies have revealed a complex interaction between Bartonella spp. Comparative Immunology, Microbiology and Infectious Diseases Volume 72, October 2020, 101515 Bartonella spp. was detected in different rodent species derived from distinct (and distant) Brazilian biomes, showing a probable dominance of this genotype. in qPCR. Microbiology is an ancient biological science in which identification and characterization of microorganisms and their interactions with the surrounding environment are studied. Rodents are reservoirs of about two-thirds of Bartonella spp. 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. Phylogenetic analyses.The sequences obtained from gltA, ftsZ, and groEL cPCR assays were identified by BLASTn (nucleotide BLAST 2.4.0 between 2 April 2016 to 15 April 2016) using megaBLAST (with default parameters) and, a posteriori, a multiple sequence alignment with sequences available in GenBank was performed using Clustal/W v. 7.0.5.3 (using default parameters) (36) and manually adjusted in BioEdit (v. 7.0.5.3) (37). The median-joining network of the gltA sequences showed a strong geographic separation between the haplotypes detected. 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. (3). existence in rodents of Turkish Thrace has been detected for the first time, and the risky habitat types were evaluated for the infection. The electropherogram quality was initially analyzed in the FinchTV 1.4.0. program (http://www.geospiza.com/ftvdlinfo.html). Trends in Microbiology 11:318–321. Microorganisms are everywhere. All of the duplicate samples with quantification cycle (Cq) values higher than 0.5 of difference were tested again. The concentration mean and absorbance ratio (260/280 nm) from the DNA spleen samples extracted were 221.6 ng/μl (ranging from 29.4 to 511.7 ng/μl; standard deviation [SD], ±130.6) and 2.1 (ranging from 1.78 to 2.25; SD, ±0.15), respectively. have adapted to rodent species. Supplemental material for this article may be found at http://dx.doi.org/10.1128/AEM.02447-16. 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. The Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) provided financial support (2015/14896-1) for this article. The numbers at the nodes correspond to bootstrap values higher than 50% accessed with 107 replicates. 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. Considering the increasing destruction of natural habitats associated with intense human activities in forests and the presence of wild rodents in peridomiciliar areas, wild rodents and humans can also share arthropod vectors and pathogens. A host is an organism that supports a parasite. have been conducted in Brazil: one in synanthropic rodents (Rattus norvegicus) in the state of Bahia (northeastern Brazil) and one in wild rodents in the state of Mato Grosso do Sul (midwestern Brazil). Definition Our understanding of cells and tissues that make up the immune system of vertebrates came from seminal studies of avian species. Although it may not reflect the natural abundance, a higher occurrence of Bartonella was observed in rodents sampled in Atlantic forest (35.9% [52/145]). Rodents are important reservoirs of different bartonellae, including some human pathogens (Birtles et al., 2001; Holmberg et al., 2003; Engbæk & Lawson, 2004; Tea et al., 2004). The tree inferred by BI as ftsZ sequences (Fig. These findings highlight the fact that even within the same species and gene (B. grahamii and gltA, respectively), different rates of mutation, recombination, and specific demographic characteristics might promote distinct haplotype diversity. Diagnostic Microbiology Micrococci are catalase-positive, oxidase-positive, strictly aerobic gram-positive cocci that grow in clusters. NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. Some studies have reported a high prevalence of infection in rodent communities, which can reach up to 82% (15). The present study aimed to (i) investigate the occurrence of Bartonella infection in wild rodents from five different Brazilian biomes by quantitative PCR (qPCR), (ii) quantify the number of DNA copies of Bartonella spp. Fifteen DNA samples positive for Bartonella spp. 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. Rodent is defined as a small mammal that has a pair of incisor teeth that are constantly growing and that are used for gnawing. Goeldi states that the paca-rana is a rodent of phlegmatic and gentle disposition,. Contrasting patterns in mammal-bacteria coevolution: Ecological fitness and strategies of adaptation of, Experimental evidence of host specificity of, Prevalence and diversity of small mammal-associated, Mixed infections, cryptic diversity, and vector-borne pathogens: evidence from, Guia dos Roedores do Brasil com chaves para gêneros baseadas em caracteres externos, Diversity and molecular characterization of novel hemoplasmas infecting wild rodents from different Brazilian biomes, Run-off replication of host-adaptability genes is associated with gene transfer agents in the genome of mouse-infecting, High prevalence and genetic heterogeneity of rodent-borne, Experimental infection of cotton rats with three natural occurring, Bartonellosis: One Health perspective for an emerging infectious disease, Submission, Review, & Publication Processes, http://mobyle.pasteur.fr/cgi-bin/MobylePortal/portal.py, Association of Bartonella Species with Wild and Synanthropic Rodents in Different Brazilian Biomes. (2019) Hof H et al. 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). 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 … The most appropriate models of nucleotide substitution (lower AIC value) were T92+G+I, GTR+G+I, TN93+G+I, and GTR+G+I for the gltA, ftsZ, groEL, and concatenated phylogenetic analyses, respectively. Any of various very numerous, mostly small mammals of the order Rodentia, having large front teeth used for gnawing. We do not retain these email addresses. MA, Mata Atlântica biome; CE, Cerrado biome; CA, Caatinga biome. Rodents play role as a reservoir for some Bartonella species which cause different clinical manifestations in humans. Gnawing; biting; corroding; applied to a destructive variety of cancer or ulcer. *Negative rodents include Gerbilliscus species, Lemniscomys species, Praomys daltoni, Praomys species, Rattus rattus, Uranomys ruddi, Nannomys minutoides/mattheyi, Myomys daltoni, Praomys rostratus, , Praomys cf. Amplification efficiency (E) was calculated from the slope of the standard curve in each run using the formula E = 10−1/slope. Animals were caught using Tomahawk and Sherman live traps during previous studies performed by the Laboratory of Trypanosomatid Biology, Laboratory of Biology and Parasitology of Wild Reservoirs Mammals, and the Laboratory of Hantaviruses and Rickettsioses, Oswaldo Cruz Institute, Rio de Janeiro, Brazil (25–27). In 1681, the Dutch microscopist Antonie van … in wild rodents from Caatinga, Cerrado, Atlantic forest, and Amazon forest biomes in Brazil. (3), which discriminate Bartonella at the species level, the answer is yes. Molecular characterization of Bartonella spp.All positive qPCR samples from rodents were submitted to the previously described conventional PCR (cPCR) assays for three other protein-coding genes, namely, gltA (350 bp), ftsZ (515 bp), and groEL (752 bp) genes (1, 17, 32, 33). 6). 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). Therefore, more studies are needed to assess the genetic diversity of Bartonella spp. described to date; and some of them have been implicated as causative agents of human diseases. Furthermore, it is important to emphasize that there is a huge overlap of distribution areas for the majority of rodent specimens found infected by Bartonella spp. In our study, we performed molecular and phylogenetic analyses of In conclusion, our study showed that the occurrence of Bartonella bacteria in rodents is much more frequent and widespread than previously recognized. 5). From the Latin prefix com- (with, together, jointly) + the Latin adjective mensalis (of the table). 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. 7th, completely revised and expanded edition. MA, Mata Atlântica biome; CE, Cerrado biome. The occurrence of Bartonella spp. In addition, our study accessed important gaps in the biology of this group of bacteria in Brazil, such as its low host specificity, high genetic diversity, and relationship with other Bartonella spp. This microscopic parasite causes a diarrheal illness in the small intestines of vertebrates called giardiasis. 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). are emerging and reemerging Gram-negative facultative intracellular Alphaproteobacteria belonging to the order Rhizobiales, family Bartonellaceae, that infect erythrocyte and endothelial cells from a wide range of animal species, including humans (1, 2). 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 humans. detected in wild rodents in Brazila. According to the genetic identity values proposed by La Scola et al. Three microliters of DNA was used as a template in 25-μl reaction mixtures containing 10× PCR buffer, 1.0 mM MgCl2, 0.6 mM deoxynucleotide triphosphate (dNTPs) mixture, 1.5 U of Taq DNA polymerase (Life Technologies), 1.25 μl of dimethyl sulfoxide (DMSO) (Sigma-Aldrich), and 0.5 μM IRBPfwd (5′-TCCAACACCACCACTGAGATCTGGAC-3′) and IRBPrev (5′-GTGAGGAAGAAATCGGACTGGCC-3′) primers. Is a rodent andisfound inconsiderable numbers in the south of Prussia. infecting wild rodents from five different Brazilian biomes. A trap intended to capture a mouse or a rat is an example of a rodent trap. In our study, we performed molecular and phylogenetic analyses of Bartonella spp. The DNA concentration and absorbance ratio (260/280 nm) were measured using a NanoDrop spectrophotometer (Thermo Scientific, Waltham, MA, USA). The haplotype diversity is controlled by different processes, including but not limited to mutation, recombination, and demography. (30) showed that a 10-fold increase in copy numbers assessed by qPCR was associated with the odds of positive results in cPCR, demonstrating the better performance of qPCR over cPCR in detecting low Bartonella DNA copy numbers (30). received a scholarship from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior. 2. Molecular identification and phylogenetic analysis. Among them, only 25 samples were simultaneously positive in gltA, ftsZ, and groEL cPCR assays. (2019) Leishmaniasis In: Hof H, Schlüter D, Dörries R, eds Duale Reihe Medizinische Mikrobiologie. Spirilla (singular, Spirillum) are a group of bacteria characterized by a corkscrew (spiral) appearance. by qPCR. 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 ]. The diversity analysis of gltA sequences showed the presence of 15 different haplotypes. The Akaike information criterion (AIC) available in MEGA 5.05 (41) was applied to identify the most appropriate model of nucleotide substitution. 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. Eighteen amplified gltA sequences shared percentages of identity ranging from 96 to 98% with Bartonella spp. Phylogenetic relationships within the Bartonella genus based on the ftsZ gene. 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. PCR amplifications were conducted in low-profile multiplate unskirted PCR plates (Bio-Rad, CA, USA) using a CFX96 thermal cycler (Bio-Rad, CA, USA). were detected in 25.6% (117/457) of rodent spleen samples analyzed, and this occurrence varied among different biomes. Bartonella bacilliformis was used as an outgroup. This question is for testing whether or not you are a human visitor and to prevent automated spam submissions. by qPCR, only 25 (21.3%) were simultaneously positive by gltA, ftsZ, and groEL cPCR assays. 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). Microbiota and its influence in animal models. 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. 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. are found widely in animals such as dogs, horses, cattle, pigs, and rodents, and are excreted in their urine. As a probable result, rodents develop a long-lasting subclinical bacteremia and a high diversity of Bartonella genotypes (13, 46–48). Bartonella henselae DNA obtained from a naturally infected cat was used as a positive control (30). 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). Topic 3. Subsequently, the sequences were submitted to phylogenetic analyses. An intimate relationship. However, limited information about the epidemiological, genetic diversity, distribution, and ecological aspects of Bartonella infections in Brazil is available as yet. Rodents were trapped using cage and break‐back traps and purchased from local markets in June and December 2006 in four Lao provinces. 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). Wild rodents are reservoirs of various zoonotic diseases, such as toxoplasmosis, babesiosis, and leishmaniasis. 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. 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. MA: Mata Atlântica; CE: Cerrado biomes. Additionally, we performed a concatenated phylogenetic analysis with the sequences which were simultaneously positive in PCR assays based on gltA, ftsZ, and groEL genes. 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. 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. A rat is an example of a rodent. Backgrounds. Rodents are reservoirs of about two-thirds of Bartonella spp. Rodents are the most diverse order of mammals with at least 2552 currently recognized species of living rodents, representing about 39.3% of all mammals. It is an anaerobic flagellated protozoan parasite. Several studies suggest that Bartonella spp. Leptospira spp. Here, we describe for the first time the prevalence and genetic properties of Bartonella organisms in wild rodents in Japan.
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