Bartonella henselae and Bartonella clarridgeiae infection, hematological changes and associated factors in domestic cats and dogs from an Atlantic rain forest area, Brazil

Accepted Manuscript Title: Bartonella henselae and Bartonella clarridgeiae infection, hematological changes and associated factors in domestic cats and dogs from an Atlantic rain forest area, Brazil Authors: Beatriz Teixeira Gomes da Silva, Aline Moreira de Souza, Sabrina Destri Emmerick Campos, Daniel de Barros Macieira, Elba Regina Sampaio de Lemos, Alexsandra Rodrigues de Mendonc¸a Favacho, N´adia Regina Pereira Almosny PII: DOI: Reference:

S0001-706X(18)31329-9 https://doi.org/10.1016/j.actatropica.2019.02.026 ACTROP 4940

To appear in:

Acta Tropica

Received date: Revised date: Accepted date:

25 October 2018 25 February 2019 26 February 2019

Please cite this article as: Silva BTGd, de Souza AM, Campos SDE, de Barros Macieira D, de Lemos ERS, de Mendonc¸a Favacho AR, Pereira Almosny NR, Bartonella henselae and Bartonella clarridgeiae infection, hematological changes and associated factors in domestic cats and dogs from an Atlantic rain forest area, Brazil, Acta Tropica (2019), https://doi.org/10.1016/j.actatropica.2019.02.026 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Bartonella henselae and Bartonella clarridgeiae infection, hematological changes and associated factors in domestic cats and dogs from an Atlantic rain forest area, Brazil. Beatriz Teixeira Gomes da Silva , Aline Moreira de Souza , Sabrina Destri Emmerick Campos , Daniel de Barros Macieira , Elba Regina Sampaio de Lemos Alexsandra Rodrigues de Mendonça Favacho , Nádia Regina Pereira Almosny

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,

Beatriz Teixeira Gomes da Silva

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Universidade Federal Fluminense, Faculdade de Veterinária, Programa de Pós-Graduação em Medicina Veterinária (Clínica e Reprodução Animal), Rua Vital Brasil Filho, 64 - Santa Rosa, Niterói - RJ, 24230-340; Rio de Janeiro, Brasil [email protected]

Aline Moreira de Souza

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Universidade Federal Fluminense, Faculdade de Veterinária, Departamento de Patologia e Clínica Veterinária, Rua Vital Brasil Filho, 64 - Santa Rosa, Niterói - RJ, 24230-340; Rio de Janeiro, Brasil [email protected]

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Sabrina Destri Emmerick Campos

Daniel de Barros Macieira

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Universidade Federal Fluminense, Faculdade de Veterinária, Programa de Pós-Graduação em Medicina Veterinária (Clínica e Reprodução Animal), Rua Vital Brasil Filho, 64 - Santa Rosa, Niterói - RJ, 24230-340; Rio de Janeiro, Brasil [email protected]

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Universidade Federal Fluminense, Faculdade de Veterinária, Departamento de Patologia e Clínica Veterinária, Rua Vital Brasil Filho, 64 - Santa Rosa, Niterói - RJ, 24230-340; Rio de Janeiro, Brasil [email protected]

Elba Regina Sampaio de Lemos

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Fundação Oswaldo Cruz (FIOCRUZ), Laboratório de Hantaviroses e Rickettsioses, Rio de Janeiro, Rio de Janeiro, Brasil [email protected]

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Alexsandra Rodrigues de Mendonça Favacho Fundação Oswaldo Cruz Mato Grosso do Sul (FIOCRUZ MS), Campo Grande, Mato Grosso do Sul, Brasil [email protected]

Nádia Regina Pereira Almosny

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Universidade Federal Fluminense, Faculdade de Veterinária, Departamento de Patologia e Clínica Veterinária, Rua Vital Brasil Filho, 64 - Santa Rosa, Niterói - RJ, 24230-340; Rio de Janeiro, Brasil [email protected]

ABSTRACT Cats are considered main reservoir of Bartonella henselae, which is transmitted to other cats especially through Ctenocephalides felis fleas, and to humans

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through scratching and biting. Serra da Tiririca State Park (PESET) is an Atlantic Forest area that shelters a wide variety of endemic fauna. Recently, the

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park has been suffering due to irregular housing construction and domestic animal population that interacts with humans and wildlife. Given that

surveillance policies for animals are part of the global Strategic Framework for

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One Health, the aim of this study was to detect Bartonella spp. DNA in cats and

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dogs, evaluating laboratory changes and associated factors. Blood samples of

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124 dogs and 89 cats were collected for hematology and serum chemistry

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analysis. DNA was extracted and tested by conventional polymerase chain reaction (PCR) targeting a fragment of the citrate synthase (gltA) gene of

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Bartonella spp. with specific primers. Positive samples were sequenced to

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identify species. Bartonella henselae and B. clarridgeiae were detected in 24.7% of cats, being, for our knowledge, the first report of B. clarridgeiae in cats

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from Rio de Janeiro, Brazil. None of the samples obtained from dogs tested positive in the PCR assays. No statistical significance was observed in physical and laboratory exams. We suggest that cats that inhabit PESET can be

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considered sources of Bartonella sp. for other cats and humans. We highlight that infected cats did not present clinical or laboratory alterations. We alert for the need of care measures, avoiding scratch and bite, particularly in immunocompromised people. Keywords: Bartonellosis, diagnosis, cat-scratch disease

1. INTRODUCTION Vector-borne diseases are globally important to human and animal health, since pathogens, vertebrate and invertebrate animals, including human

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population, may interact with each other, which have allowed contact between man and zoonotic diseases (Harrus and Beneth, 2005).

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The genus Bartonella covers a wide variety of bacterial species, and at

least 20 species cause intra-erythrocytic infections in several hosts. (Regier et al., 2016). Cats are considered main reservoir of B. henselae, B. clarridgeiae

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and B. koehlerae (Kordick et al., 1999). Transmission to cats occurs through

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vectors such as Ctenocephalis felis fleas, and to humans, transmission by

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scratching, biting or contact with saliva of cats (Tsai et al., 2011).

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Serra da Tiririca State Park (PESET) is located in Rio de Janeiro state and covers the municipalities of Niterói and Maricá. Consists of an Atlantic

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Forest area with a subtropical climate and annual average temperatures of

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23.7ºC that shelters a wide variety of rare and endemic fauna and flora. However, many private properties are located in PESET, which leads to

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increased speculation process, irregular housing construction, population increase, derogatory tourism, fire, hunting, generation of many waste and consequent irregular disposal of waste (INEA, 2012). In addition to population

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growth, there is also an increase in domestic animals population, which interact with

humans

and

wild

animals,

often

through

hunting,

increasing

pathogens´transmission risk between two populations. The global Strategic Framework for health aims to decrease the risk and minimize impact of emerging infectious diseases at animal-human ecosystem

(OIE, 2008). According to Shaffer (2008), the use of surveillance policies for animals is a part of an approach in One Health Perspective. Inspired by principles of One Health idea, which recognizes that human welfare is linked to animals and environment that surrounds him, this study aims to detect Bartonella spp. DNA in cats and dogs inhabiting PESET and to

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evaluate and compare hematological and biochemical changes and associated

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factors with positivity.

2. MATERIAL AND METHODS 2.1.

Animals

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This study was approved by the Ethics Commission on Animal Use

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(CEUA) from Universidade Federal Fluminense (UFF) under the license

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number 677. Animals included in this study were part of a free spay/neuter

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project from UFF Veterinary College, and tutors granted consent.

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Blood samples of 213 owned animals, 124 dogs and 89 cats, which lived inside PESET and its surroundings (22°56′04.28″S, 43°02′22.66″W; Niterói/RJ,

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Brazil) were collected by jugular or cephalic phlebotomy. Samples were transferred into sterile tubes containing ethylenediaminetetraacetic acid (EDTA)

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anticoagulant and tubes without anticoagulant and transported at 40C to Laboratório de Pesquisa Clínica e Diagnóstico Molecular Marcilio Dias do

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Nascimento (LAMADIN) from UFF. Animals were physically examined and the owners answered a

questionnaire about ectoparasite presence and control, oral mucosa staining, corporal score, outdoor access, contact with other animal species and hunting

habit. The evaluation of the body condition score was evaluated in the five-point system of Edney and Smith (Edney and Smith, 1986).

2.2.

Hematological and biochemical analysis

Complete blood count was processed in an automated veterinary

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hematology analyzer pocH-100iV Diff (Sysmex do Brasil Indústria e Comércio Ltda®, São José dos Pinhais, Brasil) whereas differential cell counts were

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performed manually with Diff-Quik-stained blood smears examined under an

optical microscope (E-200 Nikon) by observing 100 random microscopic fields (Magnification: 1000x) to check for the presence of Bartonella spp. inclusion.

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Serum was tested in an automated analyzer Labmax 240 Premium

gammaglutamyltransferase

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aminotransferase (ALT),

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(Labtest®, LabtestDiagnóstica AS, Lagoa Santa, Brazil) to determine alanine (GGT) and alkaline

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phosphatase (AP) activity levels, as well as urea, creatinine, total proteins and

Molecular analysis

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2.3.

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albumin levels.

DNA extraction from whole blood was performed using a commercial kit

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(illustra blood genomic Prep Mini Spin Kit; GE Healthcare Life Sciences, São Paulo, SP, Brazil). Samples were tested by conventional polymerase chain

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reaction (PCR) targeting a fragment of the citrate synthase (gltA) gene of Bartonella spp. with primers BhCS781p (5’ GGGGACCAGCTCATGGTGG 3’) and BhCS1137n (5’ AATGCAAAAAGAACAGTAA ACA 3’) (Norman et al., 1995). For each reaction, a positive control (purified DNA of B. henselae

extracted from a naturally infected cat) and a negative control (nuclease-free water) were used. Amplicons with 380 pb were visualized following electrophoresis in 1.5% agarose gel stained with GelRedTM (Uniscience, Miami, EUA) under ultraviolet (UV) light. Sensitivity of this PCR analysis was evaluated with 10serial

dilution

(109 to

10)

of

a

known

concentration

of B.

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fold

henselae DNA. Five microliters of each resulting dilution was used as a

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template for PCR under the amplification conditions described in

Materials and Methods. The limit of detection considered for this PCR was 40 copies.

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All amplification products were purified using a commercial kit (illustra

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GFX PCR DNA and Gel Band Purification Kit; GE Healthcare Life Sciences,

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São Paulo, Brazil) following the manufacturer’s instruction and sent for

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sequencing on a capillary-type Sanger platform (PDTIS/FIOCRUZ) at the

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Oswaldo Cruz Foundation, Rio de Janeiro, Brazil. In this study, we choose to work with sequences that presented fragment sizes that would permit

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calculable results when submitted to phylogenetic analyses. Partial Bartonella-gltA sequences were trimmed and assembled in a

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contig. The resulting consensus sequence was used as a query for blastn analysis against the NCBI nr database to assess sequence similarities. A

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phylogram was generated using the Neighbor Joining method. Sequence analyses and phylogenetic trees were performed in CLC Main Workbench software version 7.9.1 (QIAGEN Aarhus A/S), and bootstrap analyses with 1,000 resamplings were performed to test the robustness of the branching.

2.4.

Statistical analysis

Statistical analysis was performed using the IBM SPSS Statistic® computer program (version 21). The descriptive statistics and the Shapiro-Wilk normality test were performed for all CBC and biochemical parameters.

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Student's T-Test for independent variables was performed in variables that presented normal distribution, while those that did not present normal

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distribution were tested by the Mann-Whitney method. For the physical

examination data and the individual questionnaires, the chi-square test was used. Due to the low sample size, Fisher's Exact Test was used for all

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parameters. In all cases p-values <0.05 were considered significant. Odds

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ratios and 95% confidence intervals were provided.

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3. RESULTS

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Bartonella DNA was detected in 24.7% (22/89) of evaluated cats. None of the 124 samples obtained from dogs tested positive in the gltA PCR assays.

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Results are shown in Table 1.

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Table 1: Results obtained after PCR assays in DNA from whole blood of cats and dogs living inside the Serra da Tiririca state park and its surroundings

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(22°56′04.28″S, 43°02′22.66″W; Niterói/RJ, Brazil). PCR1 results Total Positive Negative Cat 22 67 89 Dog 0 124 124 Total 22 191 213 1Polymerase chain reaction (PCR) targeting a fragment of the citrate synthase (gltA) gene of Bartonella spp. Animal

In the present study, most of dogs presented clinically health or with nonspecific clinical manifestations. Results included 94.3% (117/124) of dogs with normal oral mucosa, 78.2% (97/124) with ideal body condition score, 66.9% (83/124) with ectoparasites (fleas, ticks or both) at the time of sampling, although 37.1% (46/124) of the owners reported the adoption of

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ectoparasite control measures. Access to outdoor environments was

notified by the owners in 68.5% (85/124) of the dogs; the hunting habit was

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present in 53.2% (66/124) of them, and 83.9% (104/124) of the dogs maintained contact with other dogs and/or cats.

Female cats accounted for 46.1% (41/89), while male cats accounted for

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53.9% (48/89). The frequency of Bartonella spp. positivity was higher among

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male cats; however, there was no statistical difference (p>0.05) between cats

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with positive and negative PCR results. Physical examination of the evaluated

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cats (n=89) revealed pale mucous membranes in 18.2% (4/22) Bartonella-

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positive cats and in 13.4% (9/67) cats with negative PCR results. Poor body condition was seen in 31.8% (7/22) infected cats and in 19.4% (13/67) non-

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infected cats. All positive cats in PCR-based assays maintained contact with other dogs and/or cats. Only 2.99% (2/67) of negative cats did not maintain

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contact with any other animal. According to cat owners, 97.8% of all studied cats (n=89) had free access to outdoor environments. Interaction with the native

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fauna of the state park and hunting habits was described in 81.8% (18/22) of Bartonella-infected cats. However, 71.6% of uninfected cats also had this profile. Regarding the presence of ectoparasites at the time of sampling, 90.9% (20/22) of positive cats were parasitized only by fleas, or by fleas and ticks. Among negative cats, this frequency was even higher (97.0%, 65/67).

Ectoparasite control measures were not adopted in 90.9% infected cats (20/22) and in 95.5% (64/67) uninfected cats. Despite these differences found between positive and negative cats, no statistical significance was observed for any of these variables. Results are described in Table 2.

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Table 2. Frequencies, p-value, odds ratio and confidence interval of clinical findings and questionnaire data from cats tested for Bartonella

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spp. infection in the Serra da Tiririca state park (PESET) and its surroundings (22°56′04.28″S, 43°02′22.66″W; Niterói/RJ, Brazil). Positive cats (n=22)

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18 (81.8%)

9 (13.4%)

4 (18.2%)

54 (80.6%)

15 (68.2%)

Body condition

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Normal (3) Below normal (1,2)

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13 (19,4%)

OR

Confidence interval

89

1.000

0.900

0.224 – 3.616

89

0.248

1.938

0.657 – 5.722

89

1.000

0.970

0.930 – 1.012

89

1.000

0.970

0.930 – 1.012

0.412

1.781

0.533 – 5.953

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Paleness

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58 (86.6%)

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Mucous membrane Normal

pvaluea

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Negative cats (n=67)

Variable

7 (31.8%)

Access to outdoor environments 65 (97.0%)

22 (100%)

2 (3%)

0

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Yes No

Contact with other animals

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Yes No

65 (97.0%)

22 (100%)

2 (3.0%)

0

Interaction with wildlife and hunting habit Yes No Ectoparasites

48 (71.6%)

18 (81.8%)

19 (28.4%)

4 (18.2%)

89

Yes

65 (97.0%)

20 (90.9%)

2 (3.0%)

2 (9.1%)

No

89

0.094

0.195

0.030 – 1.253

89

0.594

2.133

0.333 – 13.681

Ectoparasites control measures Yes No

3 (4.5%)

2 (9.1%)

64 (95.5%)

20 (90.9%)

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OR: Odds ratio; ap-values>0.05 did not differ in the Fisher's Exact Test at 5% significance

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Among naturally infected cats, the laboratory abnormalities included anemia (4.5%; 1/22), hyperproteinemia (27.3%; 6/22), and hyperglobulinemia

(18.2%; 4/22). Statistically, there was no difference in complete blood count and

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biochemical analysis between cats with and without detection of Bartonella spp.

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N

Results are shown in Tables 3 and 4.

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Table 3. Mean ± standard deviation for hematologic variables from cats tested for Bartonella spp. DNA by PCR in the Serra da Tiririca state park

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(PESET) and its surroundings (22°56′04.28″S, 43°02′22.66″W; Niterói/RJ, Brazil).

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Negative

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Variable

RBC (x106/µL)

cats (n=67)

Positive cats (n=22)

p-value*

Reference range

Mean ± SD

9.06±1.5

8.98±1.99

0.879a

5-10

12.14±1.88

11.83±1.77

0.5a

8-15

Hematocrit (%)

37.19±5.92

36.76±6.18

0.779a

24-45

MCV (fL)

41.38±3.66

41.35±2.97

0.79b

39-55

MCHC (%)

32.7±1.95

32.28±1.31

0.356b

31-35

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Hemoglobin (g/dL)

Platelets

218.41±124.82

0.743b

200-700

WBC (x103/µL)

15.67±6.32

17.16±7.59

0.349b

5.5-19.5

Basophil (/µL)

0.18±0.49

0.09±0.3

0.56b

0-2

Eosinophil (%)

6.93±5.6

6.54±3.89

0.692b

2-12

Neutrophil (%)

57.55±14.15

58.36±10.49

0.775a

35-75

Limphocyte (%)

33.75±13.05

32.77±9.74

0.712a

Monocyte (%)

1.9±1.35

2.22±1.31

0.174b

RDW-SD (fL)

33.13±5.83

33.25±3.24

RDW-CV (%)

15.73±2.52

15.88±1.86

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209.92±114.03

20-35

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1-4

0.182b 0.43b

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(x103/µL)

>0.05 did not differ at 5% significance; aStudent's T-Test; bMannWhitneytest. SD = standard deviation RBC = red blood cell count; MCV = mean corpuscular volume; MCHC = mean corpuscular hemoglobin concentration; WBC = white blood cell count; RDW-SD = red cell distribution width measured by standard deviation; RDW-CV = red cell distribution width measured by coefficient of variation.

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A

N

*p-values

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Table 4. Mean ± standard deviation biochemical variables from cats tested for Bartonella spp. DNA by PCR in the Serra da Tiririca state park (PESET)

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and its surroundings (22°56′04.28″S, 43°02′22.66″W; Niterói/RJ, Brazil).

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Negative

Variable

cats (n=67)

Positive cats (n=22)

p-value*

Reference range

Mean ± SD 42.99±19.14

52.1±8.6

0.353a

43-74

Creatinine(mg/dL)

0.99±0.27

1.04±0.23

0.391a

0.8-1.8

ALT (UI/L)

51.54±22.39

57.3±46.73

0.991b

6-83

AP (UI/L)

62.53±52.99

68.62±40.64

0.246b

25-93

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Urea (mg/dL)

GGT (UI/L)

1.45±2.83

1.81±2.58

0.334b

1.3-10

Proteins (g/dL)

7.51±0.81

7.62±0.88

0.722b

5.4-7.8

Albumin (g/dL)

2.92±0.34

2.81±0.32

0.194a

2.5-3.3

Globulins (g/dL) 4.6±0.8 4.81±0.89 0.344b 2.6-5.1 a >0.05 did not differ at 5% significance; Student's T-Test; bMannWhitneytest. SD = standard deviation ALT= alanine aminotransferase; AP = alkaline phosphatase; GGT= gammaglutamyltransferase

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*p-values

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The hematological and biochemical results of dogs were expressed

by mean ± standard deviation (SD). We observed eosinophilia (14.58% ± 9.01), neutropenia (55.28% ± 11.28), hyperproteinemia (8.3g/dL ± 1.75) with

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hyperglobulinemia (5.47g/dL ± 1.7) and high GGT levels (6.79 U/L ± 19.93).

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Normal values were obtained for red blood cells (6.4 x106/µL ± 1.13),

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Hemoglobin (13.98 g/dL ± 2.77), Hematocrit (41.96% ± 7.79), MCV (65.47fL

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± 3.90), MCHC (33.1% ± 2.48), Platelets (223.53x103/µL ± 113.38), Limphocyte (24.46% ± 10.27), Monocyte (5.6% ± 2.88), RDW-SD (35.44fL ± (11.8% ± 7.9), Urea (29.98 mg/dL ± 10.86), Creatinine

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3.36), RDW-CV

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(0.75 mg/dL ± 0.22), ALT (34.29U/L ± 18.17), ALT (34.29U/L ± 18.17), and Albumin (2.82 g/dL ± 0.57).

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Among the Bartonella-positive samples (N=22), 12 amplicons were

sequenced with success. Two species were found to infect the cats in this

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study. Phylogenetic tree is shown in Figure 1.

these

Blastn search analysis of our gltA sequences revealed that 3 of sequences

(accession

numbers

MH019295,

MH019297,

and

MH019304) showed 100% identity with B. henselae clone Lafke, obtained from cat fleas in Valdivia, Southern Chile (KY913626), B. henselae strain

BR_LHR human 346, obtained from human lymph node at Rio de Janeiro, Brazil (KT945243), and B. henselae strain M21HN, obtained from a cat during an investigation of Bartonella infection in China (FJ492802). Other 4 nucleotide sequences (accession numbers MH019298; MH019299; MH019305; MH019306) showed 99% identity with those same reference

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strains. In addition, 5 sequences (accession numbers MH019296; MH019300; MH019301; MH019302; MH019303)) were closely positioned

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(99% identity) to B. clarridgeiae clone 147_14 (KY913636) and B clarridgeiae isolate K13 (KX001761), obtained from fleas collected in cats

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at Valdívia, and cats living in Thailand, respectively.

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A multiple alignment was performed with the sequences obtained in the

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present study and species available in GenBank to generate a phylogram

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(Figure 1). Our data showed that samples grouped with B. henselae with

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bootstrap value of 89% and with B. clarridgeiae with bootstrap value of 96%.

4. DISCUSSION

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Bartonella spp. bacteremia in cats has a very large variation. A low prevalence of 1.6% (3/182) Bartonella-infected stray cats from a Center for

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Zoonosis Control (CCZ) in Mato Grosso was reported (Braga et al., 2015). In another study conducted in the same region, 2.25% (4/178) were positive for Bartonella spp. by PCR (Miceli et al., 2013). In the Brazilian Northeast region, 4.5% (9/200) cats from Maranhão were positive in PCR (Braga et al., 2012). In

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southern region of Brazil, in Rio Grande do Sul state, 17.02% (8/47) positive cats from shelters were found (Staggemeier, 2010). In the present study, the frequency of cats with molecular detection of Bartonella spp. was higher than in these other studies. On the other hand, in Rio de Janeiro state, higher prevalence had been reported among cats. Recently, 74.85 % (122/163) owned cats from Rio de

Janeiro were found to be positive for Bartonella spp. DNA. (Souza et al., 2017). Previously, 42.5% (17/40) Bartonella-infected clinically healthy cats were reported in the same region (Crissiuma et al., 2011). In Vassouras municipality, expressive 97.3% (36/37) cats from a shelter were positive for Bartonella spp by PCR (Souza et al., 2010). Probably the proximal contact between cats increased risk of transmission (Case, 2006)

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This is, for the best of our knowledge, the first report of B. clarridgeiae in cats from Rio de Janeiro. Bartonella clarridgeiae, like B. henselae, have been

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described as agents for cat scratch disease (CSD), that in immunossupressed people can evolve to endocarditis, fever of unknown origin, ophtalmological and

neurological syndromes, bacillary angiomatosis and visceral peliosis (Lamas et

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al., 2008). Domestic cat is considered source of these bacteria (Chomel et al.,

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2004).

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In this study no significative statistical difference was found in

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hematological results between Bartonella spp. positive and negative cats. Although anemia, eosinophilia, neutropenia and thrombocytopenia have

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been reported in sick animals with Bartonella spp. (Souza et al., 2017,

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Álvares-Fernández et al., 2018), mild or no abnormalities are frequently seen in Complete blood count from Bartonella infected cats.

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Among biochemical profile, mild or no abnormalities have been

reported in Bartonella infected cats and only hyperproteinemia and

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hyperglobulinemia have been reported in sick animals (Whittemore et al., 2012; Álvares-Fernández et al., 2018). No significant associations were also identified in this study between biochemical analyses and Bartonella DNA positive cats.

Souza et al. (2017) carried out a study for the detection of Bartonella spp. in domestic cats in Rio de Janeiro, finding high prevalence by PCR assays based on the htrA gene. The differences between the phylogram of the present study and that for htrA gene had low bootstrap levels, indicating that these branching structures did not have high confidence values. Regardless of how

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other Bartonella species are grouped, the phylogram (Figure 1) showed that the

species described here (B. henselae and B. clarridgeiae) were correctly

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identified and grouped with corresponding species available in the database.

It is important to note that negative results in dogs could be caused by a low bacteremia, and that Real time PCR would be a more sensitive

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method; however, it was not available that time.

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Our findings suggest that cats that inhabit PESET can be considered

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sources of Bartonella spp. in the study area for other cats and for humans. This

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is especially important because infected cats did not present clinical or

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laboratory great abnormalities. Thus, excessive care should be taken with the

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scratching and bite, mainly by immunosuppressed people.

5. CONCLUSION

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Cats can be considered as sources of Bartonella henselae and B.

clarridgeiae in the region studied and, because they do not show clinical signs,

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care with scratching and biting should be taken, mainly by immunosuppressed people. Laboratory analysis can reveal mild or no hematological and biochemical

changes,

hyperglobulinemia.

including

anemia,

hyperproteinemia

and

Clinically healthy dogs are unlikely to present bacteremia by Bartonella spp. and therefore should not be considered as potential transmitters for these bacteria. No relation was detected between hunting habits and contact with wild animals and the transmission of the bacteria. However, more studies directed to

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this interaction should be performed to confirm the absence of relation for

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Bartonella spp. transmission.

Conflict of interest

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The authors declare no competing interests.

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Acknowledgements

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To CNPq, FAPERJ and Proppi/UFF for financial support and scholarship.

A.,

Breitschwerdt,

E.B.,

Solano-Gallego,

L.,

2018.

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