Characterising the Tasmanian devil (Sarcophilus harrisii) pouch microbiome in lactating and non-lactating females.

Wildlife harbour a diverse range of microorganisms that affect their health and development. Marsupials are born immunologically naïve and physiologically underdeveloped, with primary development occurring inside a pouch. Secretion of immunological compounds and antimicrobial peptides in the epithelial lining of the female's pouch, pouch young skin, and through the milk, are thought to boost the neonate's immune system and potentially alter the pouch skin microbiome. Here, using 16S rRNA amplicon sequencing, we characterised the Tasmanian devil pouch skin microbiome from 25 lactating and 30 non-lactating wild females to describe and compare across these reproductive stages. We found that the lactating pouch skin microbiome had significantly lower amplicon sequence variant richness and diversity than non-lactating pouches, however there was no overall dissimilarity in community structure between lactating and non-lactating pouches. The top five phyla were found to be consistent between both reproductive stages, with over 85% of the microbiome being comprised of Firmicutes, Proteobacteria, Fusobacteriota, Actinobacteriota, and Bacteroidota. The most abundant taxa remained consistent across all taxonomic ranks between lactating and non-lactating pouch types. This suggests that any potential immunological compounds or antimicrobial peptide secretions did not significantly influence the main community members. Of the more than 16,000 total identified amplicon sequence variants, 25 were recognised as differentially abundant between lactating and non-lactating pouches. It is proposed that the secretion of antimicrobial peptides in the pouch act to modulate these microbial communities. This study identifies candidate bacterial clades on which to test the activity of Tasmanian devil antimicrobial peptides and their role in pouch young protection, which in turn may lead to future therapeutic development for human diseases.

Changes in the Skin Microbiota in Two Bare-nosed Wombats (Vombatus ursinus) with Differing Recovery Trajectories following Treatment for Sarcoptic Mange.

We report tracking of bacterial skin microbiota for two bare-nosed wombats (Vombatus ursinus) following in situ treatment for sarcoptic mange. Sarcoptes scabiei, the etiologic agent, has dramatic effects on skin microbiota. Our case reports show differing disease trajectory and bacterial beta diversity between the two treated individuals.

Environmental risk factors associated with the presence of Mycobacterium ulcerans in Victoria, Australia.

In recent years reported cases of Buruli ulcer, caused by Mycobacterium ulcerans, have increased substantially in Victoria, Australia, with the epidemic also expanding geographically. To develop an understanding of how M. ulcerans circulates in the environment and transmits to humans we analyzed environmental samples collected from 115 properties of recent Buruli ulcer cases and from 115 postcode-matched control properties, for the presence of M. ulcerans. Environmental factors associated with increased odds of M. ulcerans presence at a property included certain native plant species and native vegetation in general, more alkaline soil, lower altitude, the presence of common ringtail possums (Pseudocheirus peregrinus) and overhead powerlines. However, only overhead powerlines and the absence of the native plant Melaleuca lanceolata were associated with Buruli ulcer case properties. Samples positive for M. ulcerans were more likely to be found at case properties and were associated with detections of M. ulcerans in ringtail possum feces, supporting the hypothesis that M. ulcerans is zoonotic, with ringtail possums the strongest reservoir host candidate. However, the disparity in environmental risk factors associated with M. ulcerans positive properties versus case properties indicates the involvement of human behavior or the influence of other environmental factors in disease acquisition that requires further study.

Koala cathelicidin PhciCath5 has antimicrobial activity, including against Chlamydia pecorum.

Devastating fires in Australia over 2019-20 decimated native fauna and flora, including koalas. The resulting population bottleneck, combined with significant loss of habitat, increases the vulnerability of remaining koala populations to threats which include disease. Chlamydia is one disease which causes significant morbidity and mortality in koalas. The predominant pathogenic species, Chlamydia pecorum, causes severe ocular, urogenital and reproductive tract disease. In marsupials, including the koala, gene expansions of an antimicrobial peptide family known as cathelicidins have enabled protection of immunologically naïve pouch young during early development. We propose that koala cathelicidins are active against Chlamydia and other bacteria and fungi. Here we describe ten koala cathelicidins, five of which contained full length coding sequences that were widely expressed in tissues throughout the body. Focusing on these five, we investigate their antimicrobial activity against two koala C. pecorum isolates from distinct serovars; MarsBar and IPTaLE, as well as other bacteria and fungi. One cathelicidin, PhciCath5, inactivated C. pecorum IPTaLE and MarsBar elementary bodies and significantly reduced the number of inclusions compared to the control (p<0.0001). Despite evidence of cathelicidin expression within tissues known to be infected by Chlamydia, natural PhciCath5 concentrations may be inadequate in vivo to prevent or control C. pecorum infections in koalas. PhciCath5 also displayed antimicrobial activity against fungi and Gram negative and positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). Electrostatic interactions likely drive PhciCath5 adherence to the pathogen cell membrane, followed by membrane permeabilisation leading to cell death. Activity against E. coli was reduced in the presence of 10% serum and 20% whole blood. Future modification of the PhciCath5 peptide to enhance activity, including in the presence of serum/blood, may provide a novel solution to Chlamydia infection in koalas and other species.

Clinical ophthalmological diagnostic description of 10 healthy sugar gliders (Petaurus breviceps) and prevalence of ocular-related presentations in a larger hospital population.

OBJECTIVE: To provide reference values for ocular examination and diagnostics in ophthalmologically normal sugar gliders (Petaurus breviceps). To retrospectively determine the prevalence of ocular diseases in sugar gliders presenting to a single institution. ANIMALS: Ten client owned and 106 previously evaluated sugar gliders. PROCEDURE: A descriptive study evaluated sugar gliders presented to Colorado State University's Avian, Exotics, and Zoological Medicine Service (CSU-AEZ) from August-2019 to January-2020. A complete ophthalmic examination including Schirmer tear test II (STT II), phenol red threat test (PRTT), intraocular pressure (IOP) via rebound tonometry, fluorescein, and rose bengal stain was performed under anesthesia. Conjunctival aerobic culture swabs and cytology were collected prior to ophthalmic evaluation. A retrospective review of medical records of sugar gliders presented to CSU-AEZ from 2008 to 2018 for ocular disease was performed. RESULTS: Mean values ± standard deviation for selected diagnostics included the following: STT II: 2.2 ± 6.7 mm/min; PRTT: 0 ± 0 mm/15 s; IOP: 12 ± 2.6 mm Hg. Fluorescein and rose bengal staining highlighted corneal abrasions secondary to tear testing. The three most common conjunctival bacterial isolates cultured were Staphylococcus spp. (3/20, 15%), Coryneform spp. (3/20, 15%), and unidentified Gram-positive cocci (3/20, 15%). Retrospective analysis revealed ocular diseases to be the third most common abnormality resulting in sugar glider presentations (13/106, 12.3%). CONCLUSION: This descriptive study gives reference values for IOP, conjunctival microbiology, and cytology for sugar gliders. STT II and PRTT provide little clinical value in sugar gliders. The retrospective study revealed that ocular abnormalities, often secondary to dental disease, are a common reason for presentation.

Low occurrence of Bartonella in synanthropic mammals and associated ectoparasites in peri-urban areas from Central-Western and Southern Brazil.

Worldwide, Bartonella species are known to infect a wide range of mammalian and arthropod hosts, including humans. The current study aimed to investigate the prevalence of Bartonella spp. in synanthropic mammals captured in peri-urban areas from Central-Western and Southern Brazil and their ectoparasites. For this aim, 160 mammals belonging to four species, and 218 associated arthropods were sampled. DNA was extracted and subjected to different Bartonella screening assays. Additionally, blood samples from 48 small rodents were submitted to liquid BAPGM culture followed by qPCR assay and solid culture. Two out of 55 Rattus captured in Santa Catarina state were PCR-positive for Bartonella when targeting the nuoG, 16S, and ITS loci. Sequences showed high homology with Bartonella coopersplainsensis. Conversely, all 48 small rodents, 14 capybaras and 43 opossum DNA samples from animals trapped in Mato Grosso do Sul were Bartonella negative in the HRM real time PCR assays targeting the ITS locus and gltA gene. Additionally, all mammal-associated ectoparasites showed negativity results based on HRM real time PCR assays. The present study showed, for the first time, the occurrence of B. coopersplainsensis in Brazil, shedding some light on the distribution of rats-related Bartonella in South America. In addition, the majority of rodents and marsupials were negative for Bartonella spp. Since B. coopersplainsensis reservoirs - Rattus spp. - are widely dispersed around the globe, their zoonotic potential should be further investigated.

Phase variation in latB associated with a fatal Pasteurella multocida outbreak in captive squirrel gliders.

A septicaemic disease outbreak caused by Pasteurella multocida at a zoo in Western Australia (Zoo A) occurred in a resident group of squirrel gliders (Petaurus norfolcensis) following the introduction of two squirrel gliders imported from another zoo (Zoo B). P. multocida isolates obtained from the affected animals and asymptomatic, cohabiting marsupials at both zoos were typed via lipopolysaccharide outer core biosynthesis locus (LPS) typing, repetitive extragenic palindromic PCR (Rep-PCR) typing, and multilocus sequence typing (ST). Investigation of isolate relatedness via whole genome sequencing (WGS) and phylogenomic analysis found that the outbreak isolates shared the same genetic profile as those obtained from the imported gliders and the positive marsupials at Zoo B. Phylogenomic analysis demonstrated that these isolates belonged to the same clone (named complex one), confirming that the outbreak strain originated at Zoo B. As well, the carriage of multiple different strains of this pathogen in a range of marsupials in a zoo setting has been demonstrated. Importantly, the genomic investigation identified a missense mutation in the latB, a structural LPS gene, resulting in introduction of an immediate stop codon in the isolates carried by asymptomatic squirrel gliders in Zoo B. The identified diversity in the latB gene of LPS outer core biosynthesis loci of these isolates is consistent with a novel phase variable mechanism for virulence in P. multocida. Our study demonstrates the benefit of WGS and bioinformatics analysis in epidemiological investigations of pasteurellosis and its potential to reveal unexpected insights into bacterial virulence.

Microbiological survey of sugar gliders (Petaurus breviceps) kept as pets in Italy.

The sugar glider (Petaurus breviceps) is a small, arboreal, nocturnal, gliding mammalian possum belonging to the marsupial infraclass. Exotic marsupials, including sugar gliders, are becoming popular companion pets and, consequently, the risk of potential infections that can be transmitted to humans should be investigated. Data on the role of the sugar glider as a possible carrier of pathogenic and zoonotic bacteria are scarce and fragmentary. Therefore, this study is aimed at evaluating the prevalence of potentially zoonotic bacteria (Salmonella spp., Escherichia coli, Campylobacter spp., Pseudomonas spp., Klebsiella spp., Listeria monocytogenes and Yersinia enterocolitica) in 64 sugar gliders kept as pets in Italy. The highest prevalence of infection pertained to members of the family Enterobacteriaceae, in particular Citrobacter spp. (50%), Enterobacter spp. (28·1%) and Klebsiella pneumoniae (15·6%); Pseudomonas aeruginosa was isolated from 10 out of 64 samples (15·6%). All strains of Klebsiella pneumoniae exhibited some level of resistance to multiple antimicrobials (ampicillin, amoxicillin-clavulanic acid and doxycycline). SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study show that sugar gliders may act as carriers of potentially pathogenic agents for humans and other animal species, therefore caution should be exercised in the handling and contact with these animals.

Ongoing quiescence in the Borborema Plateau Plague focus (Paraiba, Brazil).

Plague is a zoonosis caused by Yersinia pestis, whose cycle is based on a reservoir system composed of mammals and their fleas. Its transmission cycle presents long enzootic periods with undetected cases, sometimes misleading that the cycle is extinct. While surveillance activities in Brazil are being carried out only in some focal areas, the serologic results confirm the persistence of Y. pestis in all monitored areas. We studied the small mammal assembly and Y. pestis presencein the Borborema Plateau Focus within the state of Paraíba, which staged the last Brazilian plague outbreak (1986-1987), through aninventory and Y. pestis detection survey of small mammals in peridomestic and sylvatic areas from two municipalities in the state of Paraíba.The field sampling captured 45 specimens (27 marsupials, 18 rodents), of 10 species. Only two species (one marsupial, one rodent) were captured in both peridomestic and sylvatic ecotopes. The sylvatic ecotope had higher richness and abundance. No evidence of circulation of the pathogen was detected, however, this result does not discard the necessity of continuous epidemiological surveillance due to the risk of rekindling the foci after long dormant periods, especially given the current epidemiological transition occurring on a Global scale.

Detection of antibodies against Leptospira spp in free-living marsupials caught in the Eastern Amazon.

INTRODUCTION: Serological surveys are important to assess the health status of wild animals. In this study, antibodies against Leptospira spp, causal agents of leptospirosis, were detected in free-living marsupials in the State of Pará, Brazil. METHODS: Nineteen blood samples collected from marsupials in the municipalities of Peixe-Boi, Viseu, and Castanhal were subjected to microscopic agglutination tests. RESULTS: In total, 36.8% (7/19) of samples were positive, and two exhibited co-agglutination. The most frequent serovars were Icterohaemorrhagiae (60%; 3/5), Panama (20%; 1/5), and Nupezo (20%; 1/5). CONCLUSIONS: Anti-Leptospira spp antibodies currently circulate in free-living marsupials in Northeastern Pará.

Detection of antibodies against Leptospira spp in free-living marsupials caught in the Eastern Amazon

Abstract INTRODUCTION: Serological surveys are important to assess the health status of wild animals. In this study, antibodies against Leptospira spp, causal agents of leptospirosis, were detected in free-living marsupials in the State of Pará, Brazil. METHODS: Nineteen blood samples collected from marsupials in the municipalities of Peixe-Boi, Viseu, and Castanhal were subjected to microscopic agglutination tests. RESULTS: In total, 36.8% (7/19) of samples were positive, and two exhibited co-agglutination. The most frequent serovars were Icterohaemorrhagiae (60%; 3/5), Panama (20%; 1/5), and Nupezo (20%; 1/5). CONCLUSIONS: Anti-Leptospira spp antibodies currently circulate in free-living marsupials in Northeastern Pará.

Mycobacterium ulcerans DNA in Bandicoot Excreta in Buruli Ulcer-Endemic Area, Northern Queensland, Australia.

To identify potential reservoirs/vectors of Mycobacterium ulcerans in northern Queensland, Australia, we analyzed environmental samples collected from the Daintree River catchment area, to which Buruli ulcer is endemic, and adjacent coastal lowlands by species-specific PCR. We detected M. ulcerans DNA in soil, mosquitoes, and excreta of bandicoots, which are small terrestrial marsupials.

Molecular evidence of Chlamydia pecorum and arthropod-associated Chlamydiae in an expanded range of marsupials.

The order Chlamydiales are biphasic intracellular bacterial pathogens infecting humans and domesticated animals. Wildlife infections have also been reported, with the most studied example being Chlamydia pecorum infections in the koala, an iconic Australian marsupial. In koalas, molecular evidence suggests that spill-over from C. pecorum infected livestock imported into Australia may have had a historical or contemporary role. Despite preliminary evidence that other native Australian marsupials also carry C. pecorum, their potential as reservoirs of this pathogen and other Chlamydia-related bacteria (CRBs) has been understudied. Mucosal epithelial samples collected from over 200 native Australian marsupials of different species and geographic regions across Australia were PCR screened for Chlamydiales. Previously described and genetically distinct C. pecorum genotypes and a range of 16S rRNA genotypes sharing similarity to different CRBs in the broader Chlamydiales order were present. One 16S rRNA Chlamydiales genotype recently described in Australian ticks that parasitise native Australian marsupials was also identified. This study provides further evidence that chlamydial infections are widespread in native fauna and that detailed investigations are required to understand the influence these infections have on host species conservation, but also whether infection spill-over plays a role in their epidemiology.

Lactococcus petauri sp. nov., isolated from an abscess of a sugar glider.

A strain of lactic acid bacteria, designated 159469T, isolated from a facial abscess in a sugar glider, was characterized genetically and phenotypically. Cells of the strain were Gram-stain-positive, coccoid and catalase-negative. Morphological, physiological and phylogenetic data indicated that the isolate belongs to the genus Lactococcus. Strain 159469T was closely related to Lactococcus garvieae ATCC 43921T, showing 95.86 and 98.08 % sequence similarity in 16S rRNA gene and rpoB gene sequences, respectively. Furthermore, a pairwise average nucleotide identity blast (ANIb) value of 93.54 % and in silico DNA-DNA hybridization value of 50.7  % were determined for the genome of strain 159469T, when compared with the genome of the type strain of Lactococcus garvieae. Based on the data presented here, the isolate represents a novel species of the genus Lactococcus, for which the name Lactococcus petauri sp. nov. is proposed. The type strain is 159469T (=LMG 30040T=DSM 104842T).

New Trypanosoma species, Trypanosoma gennarii sp. nov., from South American marsupial in Brazilian Cerrado.

Hundreds of trypanosome species have been described in all mammalian orders, on every continent, including with mixed infections. Trypanosomes circulate in the form of sylvatic enzootic infections transmitted by blood-sucking insects that are associated with the host mammals. Small wild mammals were caught in a fragment of Cerrado terrain on an island in the hydroelectric reservoir of Três Marias, in the central region of the state of Minas Gerais, using pitfall and Sherman traps with different means of attraction. DNA samples from these mammals were subjected to the conventional polymerase chain reaction (PCR) for the full-length genes SSU rDNA and gGAPDH. A total of 232 animals of the orders Didelphimorphia, Rodentia, Chiroptera and Cingulata were caught (total of 17 species). There were also four species of marsupials: Monodelphis domestica, Didelphis albiventris, Gralicinanus agilis and Micoureus paraguaianus. Among these, there were eight positive individuals of Monodelphis domestica. However, nine cultures were established, because one of them was parasitized by two species of trypanosomes: Trypanosoma cruzi and a new trypanosome species. The new species have a large epimastigote forms, and with a well-developed undulating membrane in trypomastigote forms. The new species Trypanosoma gennarii was described in Monodelphis domestica.

Francisella tularensis ssp. holarctica in Ringtail Possums, Australia.

The occurrence of Francisella tularensis outside of endemic areas, such as North America and Eurasia, has been enigmatic. We report the metagenomic discovery and isolation of F. tularensis ssp. holarctica biovar japonica from diseased ringtail possums in Sydney, Australia. This finding confirms the presence of F. tularensis in the Southern Hemisphere.

Rickettsia amblyommii associado a roedores e marsupiais nativos da Estação Experimental Rafael Fernandes da UFERSA, Rio Grande do Norte / Rickettsia amblyommii associated with rodent and marsupials native of Raphael Fernandes Experimental Station da UFERSA, Rio Grande do Norte, Brazil

O presente estudo teve como objetivo registrar a ocorrência de Rickettsia sp. em roedores e marsupiais nativos da Estação Experimental Rafael Fernandes da UFERSA, Mossoró/RN. O trabalho consistiu em uma pesquisa de campo, com roedores e marsupiais silvestres, com os dados expressos em frequência simples e porcentagem através do programa estatístico IBM SPSS (Armonk, NY: IBM Corp.), versão 22.0. Coletaram-se amostras de plasma sanguíneo de marsupiais (36) e de roedores (5). Destes, 64 continham Amblyomma auricularium, 7 Amblyomma parvum e 12 Amblyomma sp. As amostras de plasma sanguíneo foram analisadas através da técnica de Reação de Imunofluorescência Indireta. Exemplares de A. auricularium e a A. parvum foram macerados e submetidos a Técnica de Reação em Cadeia da Polimerase. Das amostras de plasma testadas, 17,60% apresentaram soropositividade para Rickettsia amblyommii. Oito exemplares de A. auricularium estavam positivos para R. amblyommii na análise de fragmentos dos genes gltA (350 bp) e ompA (587 pb), com 100% de similaridade com Candidatus R. amblyommii estirpe Bahia e AaPE, corres­pondendo a uma baixa circulação do agente dentre os vetores e hospedeiros. Esta pesquisa registra pela primeira vez a ocorrência de R. amblyommii em marsupiais Gracilinanus agilis e Monodelphis domestica pertencentes a Família Didelphidae, e roedores das Famílias Echimyidae e Cricetidae, cujas espécies foram Thrichomys sp. e Wiedomys sp., respectivamente, em Mossoró, estado do Rio Grande do Norte.(AU)

The study aimed to register the occurrence of Rickettsia sp. in rodents and marsupials native of the Rafael Fernandes Experimental Station of UFERSA, Mossoró/RN, Brazil. The study consisted of field research on small wild mammals, with data expressed in simple frequency and percentage through IBM SPSS (IBM Corp., Armonk, NY), version 22.0. Samples of blood plasma from 36 marsupials and 5 rodents were collected. From these, 64 contained Amblyomma auricularium, 7 Amblyomma parvum and 12 Amblyomma sp. All blood plasma samples were analyzed by indirect immunofluorescence technique, and 16 macerated specimens of A. auricularium and 3 of A. parvum were analyzed by reaction technique Polymerase Chain. From the tested plasma samples 17.60% were seropositive for Rickettsia amblyommii, 8 were positive for A. auricularium e R. amblyommii in gene gltA analysis of the fragments (350 bp) and ompA (587 bp) with 100% similarity with Candidatus R. amblyommii Bahia and AAPE strain, what corresponded to a low circulation of the agent from the vectors and hosts. This study registers for the first time the occurrence of R. amblyommii in marsupials Gracilinanus agilis and Monodelphis domestica belonging to the Didelphidae family, and in rodents of the Echimyidae and Cricetidae families, the species of which were Thrichomys sp. and Wiedomys sp. respectively, in Mossoró, Rio Grande do Norte.(AU)

Bartonella species pathogenic for humans infect pets, free-ranging wild mammals and their ectoparasites in the Caatinga biome, Northeastern Brazil: a serological and molecular study

ABSTRACT This study verified the occurrence of Bartonella spp. in dogs, cats, wild mammals and their ectoparasites in Petrolina and Lagoa Grande Counties, Pernambuco, located in a semi-arid region in Northeastern Brazil. Anti-Bartonella spp. antibodies were detected by indirect immunofluorescence assay (IFA) in 24.8% of dogs (27/109) and in 15% of cats (6/40). Bartonella sp. DNA was identified by PCR performed on DNA extracted from blood and ectoparasites using primers targeting Bartonella sp. gltA and ribC genes in 100% (9/9) of Pulex irritans from Cerdocyon thous, 57.4% (35/61) of P. irritans from dogs, 2.3% (1/43) of Ctenocephalides felis felis from dogs, 53.3% (24/45) of C. felis felis from cats, and 10% (1/10) of Polyplax spp. from Thrichomys apereoides. DNA sequencing identified Bartonella clarridgeiae and Bartonella henselae in C. felis felis from cats, Bartonella rochalimae in P. irritans from dog and C. thous, and Bartonella vinsoni berkhofii in P. irritans from dog.

Occurrence and molecular characterization of hemoplasmas in domestic dogs and wild mammals in a Brazilian wetland.

Hemotropic mycoplasmas are known to cause anemia in several mammalian species. The present work aimed to investigate the occurrence of Mycoplasma spp. in wild mammals, domestic dogs and their respective ectoparasites, in southern Pantanal region, central-western Brazil. Between August 2013 and March 2015, 31 Nasua nasua, 78 Cerdocyon thous, seven Leopardus pardalis, 42 dogs, 110 wild rodents, and 30 marsupials were trapped and ectoparasites (ticks and fleas) found parasitizing the animals were collected. Mammals and ectoparasites DNA samples were submitted to conventional PCR assays for Mycoplasma spp. targeting 16S rRNA and RnaseP genes. Twenty-four N. nasua, three C. thous, two domestic dogs, one L. pardalis and one wild rodent were positive for 16S rRNA PCR protocols. Fourteen N. nasua samples were also positive in RnaseP PCR. No marsupial or arthropod showed positivity for Mycoplasma spp. The phylogenetic analyses based on 16S rRNA gene showed that all sequences obtained from dogs, two sequences obtained from C. thous and ten sequences obtained from N. nasua showed to be closely related to Mycoplasma haemocanis/Mycoplasma haemofelis species. Genotypes closely related to 'Candidatus Mycoplasma haemominutum' and Mycoplasma haemomuris were detected in the L. pardalis and in the wild rodent, respectively. Probably a novel Mycoplasma genotype, closely related to a sequence obtained from a Brazilian capybara was detected in 14 N. nasua, based on a concatenated phylogenetic analysis of 16S rRNA and RnaseP genes. The present study revealed that wild animals in southern Pantanal region, Brazil, are exposed to different species of hemoplasmas.

Bartonella species pathogenic for humans infect pets, free-ranging wild mammals and their ectoparasites in the Caatinga biome, Northeastern Brazil: a serological and molecular study.

This study verified the occurrence of Bartonella spp. in dogs, cats, wild mammals and their ectoparasites in Petrolina and Lagoa Grande Counties, Pernambuco, located in a semi-arid region in Northeastern Brazil. Anti-Bartonella spp. antibodies were detected by indirect immunofluorescence assay (IFA) in 24.8% of dogs (27/109) and in 15% of cats (6/40). Bartonella sp. DNA was identified by PCR performed on DNA extracted from blood and ectoparasites using primers targeting Bartonella sp. gltA and ribC genes in 100% (9/9) of Pulex irritans from Cerdocyon thous, 57.4% (35/61) of P. irritans from dogs, 2.3% (1/43) of Ctenocephalides felis felis from dogs, 53.3% (24/45) of C. felis felis from cats, and 10% (1/10) of Polyplax spp. from Thrichomys apereoides. DNA sequencing identified Bartonella clarridgeiae and Bartonella henselae in C. felis felis from cats, Bartonella rochalimae in P. irritans from dog and C. thous, and Bartonella vinsoni berkhofii in P. irritans from dog.