Characterizing the blood microbiota in healthy and febrile domestic cats via 16s rRNA sequencing.

This study aimed to evaluate the blood bacterial microbiota in healthy and febrile cats. High-quality sequencing reads from the 16S rRNA gene variable region V3-V4 were obtained from genomic blood DNA belonging to 145 healthy cats, and 140 febrile cats. Comparisons between the blood microbiota of healthy and febrile cats revealed dominant presence of Actinobacteria, followed by Firmicutes and Proteobacteria, and a lower relative abundance of Bacteroidetes. Upon lower taxonomic levels, the bacterial composition was significantly different between healthy and febrile cats. The families Faecalibacterium and Kineothrix (Firmicutes), and Phyllobacterium (Proteobacteria) experienced increased abundance in febrile samples. Whereas Thioprofundum (Proteobacteria) demonstrated a significant decrease in abundance in febrile. The bacterial composition and beta diversity within febrile cats was different according to the affected body system (Oral/GI, systemic, skin, and respiratory) at both family and genus levels. Sex and age were not significant factors affecting the blood microbiota of febrile cats nor healthy ones. Age was different between young adult and mature adult healthy cats. Alpha diversity was unaffected by any factors. Overall, the findings suggest that age, health status and nature of disease are significant factors affecting blood microbiota diversity and composition in cats, but sex is not.

Genetic diversity of Bartonella rpoB haplotypes in domestic cats from Chile.

The study aimed to determine the inter and intra-host Bartonella spp. genetic diversity in cats from Chile. 'Seventy-nine cats' blood DNA samples qPCR Bartonella spp. positive were subjected to T-A cloning of Bartonella spp. rpoB partial gene (825 bp), and sequencing by Sanger method. The sequences were submitted to phylogenetic and polymorphism analysis. Thirty-six (45.6%) samples were successfully cloned, generating 118 clones of which 109 showed 99.6%-100% identity with Bartonella henselae whereas 9 showed 99.8-100% identity with Bartonella koehlerae. Haplotype analysis yielded 29 different rpoB-B. henselae haplotypes, one (hap#2) overrepresented in 31 out of 33 cats, and 4 rpoB-B. koehlerae haplotypes, with hap#2 represented in all 3 B. koehlerae infected cats. More than one rpoB -B. henselae and B. koehlerae haplotypes were identified in individual cats, reporting by first time coinfection by different B. henselae/B. koehlerae rpoB variants in cats from Chile.

Molecular survey and phylogenetic analysis of Bartonella sp., Coxiella sp., and hemoplamas in pudu (Pudu puda) from Chile: first report of Bartonella henselae in a wild ungulate species.

Introduction: Recent evidence shows a high diversity of infectious agents in wildlife that represent a threat to human, domestic, and wild animal health. In Chile, wild populations of the most common cervid species, pudu (Pudu puda), have been reported as hosts for novel pathogens such as Mycoplasma ovis-like and a novel ecotype of Anaplasma phagocytophilum. A better understanding of the epidemiology of this group and other intracellular bacteria that might have cervids as hosts would enlighten their population relevance. This study aimed to determine the occurrence and genetic diversity of Bartonella spp., hemotropic mycoplasmas, and Coxiella burnetii in pudus from Chile. Methods: The DNA was extracted from the blood samples of 69 wild free-ranging and 30 captive pudus from Chile. A combination of real-time (nouG gene for Bartonella and IS1111 element for C. burnetii) and conventional PCR (16S rRNA for hemotropic Mycoplasma spp. and rpoB, gltA, and ITS for Bartonella spp.) was used for pathogen screening and molecular characterization. Results: DNA of Bartonella spp. was detected in 10.1% [95% CI (5.2-18.2%)] samples, hemotropic Mycoplasma spp. in 1.7% [95% CI (0.08-10.1%)], and C. burnetii in 1.0% [95% CI (0.05-6.3%)] samples. Two sequenced samples were identified as Mycoplasma ovis-like, and one free-ranging pudu was positive for C. burnetii. While one captive and two free-ranging pudus were positive for Bartonella henselae, one wild pudu was co-positive for B. henselae and Bartonella sp., similar to Bartonellae identified in ruminants. Discussion: To the best of our knowledge, this is the first report of B. henselae in wild ungulate species, and C. burnetii and Bartonella spp. in wild ungulate species in South America. Further research will be necessary to evaluate the potential role of pudu as reservoirs of infection and identify the sources for disease transmission among humans and wild and domestic animals.

Molecular detection and characterization of Bartonella spp. in rodents from central and southern Chile, with emphasis on introduced rats (Rattus spp.).

Bartonella spp. was screened in 155 rodents from Chile, mainly the invasive rats Rattus norvegicus and Rattus rattus. A total of 155 spleen and 50 blood samples were analyzed through real-time PCR for Bartonella spp. (nuoG gene). Positive samples were subjected to amplification of fragment of loci gltA, rpoB and ITS by conventional PCR (cPCR). Overall, 43 spleen samples (27.7%) and 6 rodent blood samples (12%) were positive for nuoG-Bartonella spp. Positive samples were found in R. norvegicus, R. rattus, Abrothrix olivacea and Oligoryzomys longicaudatus. Bartonella spp. DNA was amplified by cPCR in 16 samples, resulting in 21 sequences (6 gltA, 5 ITS and 10 rpoB). Sequencing and phylogenic analyses identified genotypes from Rattus spp., potentially belonging to Bartonella coopersplainsensis, Bartonella henselae, Bartonella tribocorum, and an undescribed Bartonella sp. From native rodents, one sequence was identified, being related B. machadoae. In conclusion, this work describes diverse and potentially zoonotic Bartonella spp. genotypes in Rattus spp. Additionally, this is the first report of Bartonella in O. longicaudatus, including a potentially novel Bartonella genotype or species.

Bartonella spp. in households with cats: Risk factors for infection in cats and human exposure.

The aim of this study was to estimate the occurrence of Bartonella spp. per household in cats and the risk factors for Bartonella spp. positivity in cats and their owners from Valdivia, Chile. A total of 464 cats (distributed within 324 households) and 326 humans (control group [n = 112] and cat owner [n = 214]) distributed in 262 households were sampled. From the cat owners (n = 214), 128 humans were in households where the cat was also sampled, totaling 84 households with dual sampling. Real-time PCR (qPCR) was used for Bartonella spp. detection in blood from cats and humans, and immunofluorescent immunoassay (IFA) anti-Bartonella henselae was performed in human serum samples. Out of the total of 324 households, 20.43% presented at least one Bartonella positive cat. From the households with dual sampling, 29.7% (25/84) presented at least one qPCR-Bartonella spp. positive cat. However, Bartonella DNA was not amplified in humans, and in 7.3% (6/82) of the households was found at least one of the cat's owners exposed to B. henselae. Cats younger than one year (Odds Ratio (OR) = 5.3), non-neutered (OR 3.46), sampled at home (OR 5.82), and with improper application of tick/flea control products (OR 3.13) showed a higher risk for Bartonella spp. presence. Humans with occupational exposure involving animal contact, were more likely to exhibit B. henselae seropositivity (OR 7.5). Bartonella spp. was present in the cats a moderate number of households, but Bartonella DNA was not detected in owners' blood, inferring that there is a low risk of recent human infection in the studied population.

Molecular identification of Bartonella spp. and Rickettsia felis in fox fleas, Chile.

Seventy-five flea pools (one to ten fleas per pool) from 51 Andean foxes (Lycalopex culpaeus) and five South American grey foxes or chillas (Lycalopex griseus) from the Mediterranean region of Chile were analyzed for the presence of DNA of Bartonella spp. and Rickettsia spp. through quantitative real-time PCR for the nouG and gltA genes, respectively. Positive samples were further characterized by conventional PCR protocols, targeting gltA and ITS genes for Bartonella, and gltA, ompA, and ompB genes for Rickettsia. Bartonella was detected in 48 % of the Pulex irritans pools (B. rochalimae in three pools, B. berkhoffii in two pools, B. henselae in one pool), and 8 % of the Ctenocephalides felis felis pools (B. rochalimae, one pool). Rickettsia was confirmed in 11 % of P. irritans pools and 92 % of the Ct. felis pools. Characterization confirmed R. felis in all sequenced Rickettsia-positive pools. All Ct. canis pools were negative. A Ct. felis pool from a wild-found domestic ferret (Mustela putorius furo) also resulted positive for R. felis. Although opportunistic, this survey provides the first description of zoonotic pathogens naturally circulating in fleas parasitizing Chilean free-living carnivores.

Spirocerca lupi in the stomach of two Andean foxes (Lycalopex culpaeus) from Chile.

The genus Spirocerca includes nematodes that parasitize the stomach and the oesophagus of carnivores, chiefly canids. Herein, we provide new data about the morphological, histopathological, and molecular characterization of Spirocerca sp. in Andean foxes (Lycalopex culpaeus) in Chile. Intact immature worms, identified as Spirocerca sp., were recovered in the lumen of the stomach from two foxes. Histologically, worms morphologically consistent with spirurid nematodes were present within the wall of the stomach and surrounded by nodular areas of inflammation with central necrotic debris. Molecular analysis of the cox1 gene yielded 19 sequences and 5 nucleotide sequence types with 99.95 to 99.98% similarity, being shared between both foxes. Nucleotide similarity ranged from 93.1 (with genotype 2 of S. lupi and S. vulpis) to 95.8% (with genotype 1 of S. lupi), a higher similarity than noted from sequences of S. lupi from an Andean fox from Peru (91.0 to 93.3%). However, the Poisson Tree Processes for species delineation did not support the existence of a new species Spirocerca. Phylogenetic and nucleotide analyses suggest that these specimens belong to a new variant or genotype of S. lupi or to a cryptic species. Whether the presence of the worms in the stomach has to do with genotypic differences in parasites or host or some combination is uncertain. Spirocerca lupi has never been found in Chilean dogs and must be investigated.

Molecular survey and genetic diversity of Bartonella spp. in domestic cats from Paraguay.

Although Bartonella spp. is described in cats worldwide, little is known about the occurrence and genetic diversity of Bartonella spp. in cats from South America. To date, it has only been detected in cats from Brazil, Chile and Argentina. This study aimed to undertake a molecular survey and explore the genetic diversity of Bartonella spp. in domestic cats from Paraguay. A TaqMan real-time quantitative PCR (qPCR) targeting the nuoG gene (83 bp) for Bartonella spp. was used to screen 125 blood samples from cats in Asuncion, Paraguay. nuoG qPCR-positive samples were further submitted to conventional PCR assays based on the ITS (453- 717 bp), gltA (767 bp), ftsZ (515 bp), rpoB (333 bp), ribC (585-588 bp), and pap-31 (564 bp) loci. Positive samples were sequenced for species identification, phylogenetic, and haplotype analyses. Bartonella D.N.A. was present in 20.8% (26/125) cat blood samples, with low levels of Bartonella nuoG D.N.A. cPCR products targeting gltA, ftsZ, ITS, and rpoB loci from sixteen cats were successfully sequenced. However, all nouG qPCR-positive samples were negative for the ribC and pap-31 genes. Bartonella henselae [62.5% (10/16)] and Bartonella clarridgeiae [37.5% (6/16)] were identified among the sequenced samples. Upon phylogenetic analysis, B. henselae and B. clarridgeiae from Paraguay clustered with sequences detected in domestic and wild cats, dogs, and cat fleas worldwide. Two to four haplotypes of B. henselae and B. clarridgeiae in cats from Paraguay were observed, with some being exclusive and others shared with worldwide distributed haplotypes. Here, we report B. henselae and B. clarridgeiae for the first time in cats from Paraguay. Its circulation in cats suggests the need to consider Bartonellae when testing clinical samples from suspected infectious diseases in humans from Paraguay.

Genetic diversity of Hepatozoon spp. in rodents from Chile.

This study aimed to investigate the genetic diversity of Hepatozoon spp. in rodents from Valdivia, Chile. A total of 74 rodents (synanthropic n=38; wild n=36) were trapped in Valdivia. We performed conventional PCR assays for Apicomplexa organisms targeting two overlapping 18S rDNA gene fragments (600 bp and 900 bp) followed by sequencing of selected amplicons. Hepatozoon spp. occurrence was 82.43% (61/74). Twelve sequences obtained from the 600 bp and ten from the 900 bp 18S rDNA fragments were identified as Hepatozoon sp. Six sequences obtained from 18S rDNA-based overlapping PCR protocols were used for concatenated (1,400 bp) phylogenetic, haplotype and distance analyses. Hepatozoon spp. 18S rDNA concatenated sequences from the present study were detected in Oligoryzomys longicaudatus, Rattus norvegicus, Mus musculus, and Abrothrix longipilis grouped with Hepatozoon species earlier described in rodents and reptiles from Chile and Brazil. Nucleotide polymorphism of the six 18S rDNA sequences (1,400 bp) from this study, and other Chilean sequences from rodents and rodent's ticks, showed high diversity with a total of nine Chilean haplotypes. Three haplotypes from Valdivia were identified for the first time in this study, suggesting the circulation of novel haplotypes in rodents from southern Chile.

A Molecular Survey on Neglected Gurltia paralysans and Aelurostrongylus abstrusus Infections in Domestic Cats (Felis catus) from Southern Chile.

Gurltia paralysans and Aelurostrongylus abstrusus are neglected metastrongyloid nematode species which infect domestic and wild cats in South American countries and in Chile, but no epidemiological studies on concomitant infections have been conducted in Chile so far. The aim of this study was not only to evaluate the occurrence of concomitant infections, but also to identify epidemiological risk factors associated with of G. paralysans and A. abstrusus infections in urban domestic cats (Felis catus) from Southern Chile. Blood samples from clinically healthy domestic cats from three cities of Southern Chile-Temuco, Valdivia, and Puerto Montt-were analyzed by an experimental semi-nested PCR protocol. A total of 171 apparently healthy domestic cats in Temuco (n = 68), Valdivia (n = 50), and Puerto Montt (n = 53) were sampled and analyzed. A total of 93 domestic cats (54.4%) were positive for G. paralysans, and 34 (19.9%) were positive for A. abstrusus infections. From those animals, 34 (19.9%) were co-infected. Cats positive with G. paralysans were found in all three cities; 47.2% in Puerto Montt, 48% in Valdivia, and 64.7% in Temuco. Levels of infection for A. abstrusus in the population under study were 4% (Valdivia), 10% (Puerto Montt), and 32.4% (Temuco). The present large-scale epidemiological study confirmed the presence of these neglected nematodes in domestic cat populations in Southern Chile, and described the possible risk factors associated with feline gurltiosis and aelurostrongylosis.

Molecular survey and genetic characterization of 'Candidatus Mycoplasma haemolamae' in llamas (Lama glama) and alpacas (Vicugna pacos) from Southern Chile.

This study aimed to perform a molecular survey and identification of hemotropic Mycoplasma spp. in domestic South American Camelids from Southern Chile. Conventional PCR (cPCR) for hemotropic Mycoplasma spp. based on 16S rRNA gene (620bp fragment) was performed in 87 EDTA-blood samples taken from 48 llamas (Lama glama) and 39 and alpacas (Vicugna pacos) from to Temuco, La Araucanía region and Valdivia, Los Rios region, Southern Chile. 16S rRNA hemotropic Mycoplasma PCR-positive were sequenced for species identification, phylogenetic and haplotype analyses, and further tested by cPCR targeting a fragment (160-210 bp) of the RNaseP (rnpB) gene. Based upon 16S rRNA cPCR results, the overall hemotropic Mycoplasma spp. occurrence in Southern camelids was 9.2% (8/87 [95% CI (4.0-17.3%)]), with five positive alpacas (12.8%; 5/39 [95% CI (4.3-27.4%)]) and three llamas (6.3%; 3/48 [95% CI (1.7-17.2%)]). All 16S rRNA PCR-positive samples were negative for the rnpB gene. Obtained 16S sequences presented high identity (99-100%) by BLASTn analysis to 'Candidatus Mycoplasma haemolamae' from an alpaca in the United Kingdom. Phylogenetic and haplotype analyses of the 16s rRNA gene showed high similarity among 'Candidatus M. haemolamae' sequences of this study and the ones from North America, Europe, and Asia evidencing a low diversity of Chilean samples, with only one haplotype detected (#1). Haplotype #1 from South American Camelids in Chile was worldwide distributed and observed in North America, Europe, and Asia. 'Candidatus M. haemolamae' detected for the first time in South American camelids in Southern Chile had low diversity and was worldwide spread.

Evaluation of terrestrial gastropods as possible intermediate hosts of Gurltia paralysans in southern Chile.

Gurltia paralysans is the causal agent of gurltiosis in domestic cats in South America. Although the life cycle of G. paralysans is unknown, it is thought that gastropods could act as intermediate hosts (IHs), as is the case for several nematodes in the Angiostrongylidae family. The aim of this study was to search for G. paralysans larvae in terrestrial gastropods and determine their role in the life cycle of this nematode species. Terrestrial gastropod samples (n=835) were collected in Punucapa, Valdivia, southern Chile, where cases of gurltiosis had been reported before. The samples included species from the families Arionidae, Limacidae, Helicidae and Milacidae. All gastropods were subjected to enzymatic digestion to isolate G. paralysans larvae. Ten percent of the gastropod samples were analyzed using seminested PCR targeting the 28S rRNA gene, while 2.6% were analyzed by histopathological examination. The results indicated the absence of G. paralysans when using any of the three methods. In conclusion, further studies are needed to evaluate specific species of aquatic or native gastropods acting as possible IHs (in this geographic location).

Molecular survey of Bartonella spp. and haemoplasmas in American minks (Neovison vison).

The aim of this study was to perform a molecular survey and characterize Bartonella spp. and haemotropic Mycoplasma (haemoplasmas) in invasive American minks (Neovison vison) from Southern Chile. Additionally, we addressed risk factors for positivity in both groups of agents. Blood and/or tissue samples from 246 minks were analysed by qPCR targeting the nuoG gene for Bartonella spp. and conventional (c)PCR for 16S rRNA for haemotropic Mycoplasma spp. nuoG qPCR-positive Bartonella spp. samples were submitted to cPCR assays (ITS, ribC, gltA, rpoB, pap-31 and ftsZ genes) to perform phylogenetic inferences. Haemotropic Mycoplasma spp. 16S-positive samples were further amplified by cPCR targeting RNaseP gene (160-210 bp) and by two overlapping 16S rRNA cPCR assays to amplify a larger portion of the gene (1,200bp) for phylogenetics. Bartonella DNA was detected in 8.9% of minks (22/246). Out of 22 nuoG qPCR-positive samples, one and two showed positive results in cPCR assays based on ITS and ribC, respectively. Consistent sequencing results were obtained for only one ITS sample (464 bp sequence), which shared 99.6% identity with B. clarridgeiae. Two per cent of minks (5/246) were positive for 16S rRNA haemotropic Mycoplasma-cPCR assay. Two concatenated sequences of 16S rRNA (1,176 and 1,230 bp) were obtained: one sample shared 97.87% identity with haemotropic Mycoplasma sp. from a wild rodent, and the other 96.49% identity with 'Candidatus Mycoplasma haematoparvum' from a dog. All BLAST results were supported by phylogenetic analysis. One haemoplasma RNase P sequence shared 94.86% identity with Mycoplasma haemofelis from a cat. No risk factors for PCR positivity were identified. In a nutshell, Bartonella clarridgeiae and a potentially novel haemoplasma closely related to haemoplasmas previously reported in rodents, dogs, domestic and wild cats were described for the first time in American minks.

Genetic diversity of Hepatozoon spp. in rodents from Chile / Diversidade genética de Hepatozoon spp. em roedores do Chile

Abstract This study aimed to investigate the genetic diversity of Hepatozoon spp. in rodents from Valdivia, Chile. A total of 74 rodents (synanthropic n=38; wild n=36) were trapped in Valdivia. We performed conventional PCR assays for Apicomplexa organisms targeting two overlapping 18S rDNA gene fragments (600 bp and 900 bp) followed by sequencing of selected amplicons. Hepatozoon spp. occurrence was 82.43% (61/74). Twelve sequences obtained from the 600 bp and ten from the 900 bp 18S rDNA fragments were identified as Hepatozoon sp. Six sequences obtained from 18S rDNA-based overlapping PCR protocols were used for concatenated (1,400 bp) phylogenetic, haplotype and distance analyses. Hepatozoon spp. 18S rDNA concatenated sequences from the present study were detected in Oligoryzomys longicaudatus, Rattus norvegicus, Mus musculus, and Abrothrix longipilis grouped with Hepatozoon species earlier described in rodents and reptiles from Chile and Brazil. Nucleotide polymorphism of the six 18S rDNA sequences (1,400 bp) from this study, and other Chilean sequences from rodents and rodent's ticks, showed high diversity with a total of nine Chilean haplotypes. Three haplotypes from Valdivia were identified for the first time in this study, suggesting the circulation of novel haplotypes in rodents from southern Chile.

Resumo Este estudo teve como objetivo investigar a diversidade genética de Hepatozoon spp. em roedores de Valdivia, Chile. Um total de 74 roedores (sinantrópicos n=38; selvagens n=36) foram capturados. PCR convencional foi realizada para organismos Apicomplexa, visando dois fragmentos sobrepostos do gene 18S rDNA (600 bp e 900 bp), seguida pelo sequenciamento de amplicons selecionados. A ocorrência de Hepatozoon spp. foi de 82,43% (61/74). Doze sequências obtidas dos fragmentos de 18S rDNA de 600 pb e dez dos fragmentos de 18S rDNA de 900 pb foram identificadas como Hepatozoon sp. Seis sequências obtidas, a partir de protocolos de PCR sobrepostos, foram usadas para análises filogenéticas (1.400 bp), de haplótipos e de distância. Sequências concatenadas 18S rDNA do presente estudo foram detectadas em Oligoryzomys longicaudatus, Rattus norvegicus, Mus musculus e Abrothrix longipilis e agrupadas com Hepatozoon descrito em roedores e répteis do Chile e do Brasil. A análise de polimorfismos das seis sequências deste estudo, junto com outras sequências chilenas de roedores e carrapatos de roedores, mostrou alta diversidade com um total de nove haplótipos no Chile. Três haplótipos detectados em Valdivia foram identificados pela primeira vez neste estudo, sugerindo que novos haplótipos circulam em roedores do sul do Chile.

Evaluation of terrestrial gastropods as possible intermediate hosts of Gurltia paralysans in southern Chile / Avaliação de gastrópodes terrestres como possíveis hospedeiros intermediários de Gurltia paralysans no sul do Chile

Abstract Gurltia paralysans is the causal agent of gurltiosis in domestic cats in South America. Although the life cycle of G. paralysans is unknown, it is thought that gastropods could act as intermediate hosts (IHs), as is the case for several nematodes in the Angiostrongylidae family. The aim of this study was to search for G. paralysans larvae in terrestrial gastropods and determine their role in the life cycle of this nematode species. Terrestrial gastropod samples (n=835) were collected in Punucapa, Valdivia, southern Chile, where cases of gurltiosis had been reported before. The samples included species from the families Arionidae, Limacidae, Helicidae and Milacidae. All gastropods were subjected to enzymatic digestion to isolate G. paralysans larvae. Ten percent of the gastropod samples were analyzed using seminested PCR targeting the 28S rRNA gene, while 2.6% were analyzed by histopathological examination. The results indicated the absence of G. paralysans when using any of the three methods. In conclusion, further studies are needed to evaluate specific species of aquatic or native gastropods acting as possible IHs (in this geographic location).

Resumo Gurltia paralysans é o agente etiológico da gurltiose em gatos domésticos na América do Sul. Embora o ciclo biologico de G. paralysans seja desconhecido, provavelmente é indireto com gastrópodes atuando como hospedeiros intermediários (HIs), como no caso de vários nematoides da família Angiostrongylidae. O objetivo deste estudo foi investigar a presença de larvas de G. paralysans em gastrópodes terrestres para avaliar seu papel no ciclo de vida do parasito. Amostras de gastrópodes terrestres (n = 835) foram coletadas em Punucapa, Valdivia, sul do Chile, onde casos de gurltiose foram relatados anteriormente. As amostras incluíram espécies das famílias Arionidae, Limacidae, Helicidae e Milacidae. Todos os gastrópodes foram submetidos à digestão enzimática para isolar as larvas de G. paralysans. 10% das amostras foram analisadas, utilizando-se seminested PCR para o gen 28S RNAr de G. paralysans, enquanto 2,6% foram analisados ​​por exame histopatológico. Os resultados indicaram ausência de G. paralysans em todos os três métodos. Os dados permitem concluir que são necessários mais estudos para avaliar espécies específicas de gastrópodes aquáticos ou nativos, que atuam como possíveis hospedeiros intermediários nessa localização geográfica.

Molecular Survey and Genetic Diversity of Hemoplasmas in Rodents from Chile.

Even though hemotrophic mycoplasma (hemoplasma) infections are well documented in a wide variety of hosts worldwide, there is a gap in the knowledge aobut hemoplasmas in rodents. This study aimed to molecularly survey and investigate the genetic diversity of hemoplasmas in rodents from Chile. Synanthropic and wild rodents (n = 74) were captured in the southern province of Valdivia (Corral, Valdivia, Riñihue, and Reumén localities). Spleen samples were submitted to a conventional PCR for hemotrophic Mycoplasma spp. targeting the 16S rRNA gene (800 bp), followed by sequencing, phylogenetic, and genetic diversity analyses. The overall occurrence of hemotrophic mycoplasmas in rodents from Valdivia was 24.5% (18/74) [95% CI (14.5; 34.1)]. Hemoplasmas were detected in Mus musculus (1/4), Rattus norvegicus (1/16), Abrothrix longipilis (7/13), A. olivaceo (6/8), and Oligoryzomys longicaudatus (3/10). The nucleotide polymorphism analysis of the targeted 16S rRNA region showed low diversity, with two genotypes and a high identity to the variants detected in wild rodents from Brazil. Hemoplasmas are described for the first time in rodents from Chile with a moderate occurrence and low 16S rDNA genetic diversity within the sampled rodent population. The detected hemoplasma genotypes were specific to rodents and were not shared with other mammals.