Molecular characterization and phylogenetic analysis of the causative agent of hemoplasma infection in small Indian Mongoose (Herpestes Javanicus)

G Model

ARTICLE IN PRESS

CIMID-970; No. of Pages 5

Comparative Immunology, Microbiology and Infectious Diseases xxx (2014) xxx–xxx

Contents lists available at ScienceDirect

Comparative Immunology, Microbiology and Infectious Diseases journal homepage: www.elsevier.com/locate/cimid

Molecular characterization and phylogenetic analysis of the causative agent of hemoplasma infection in small Indian Mongoose (Herpestes Javanicus) Hassan Sharifiyazdi a , Saeed Nazifi a,∗ , Hesamaddin Shirzad Aski b , Hossein Shayegh c a b c

Department of Clinical Sciences, School of Veterinary Medicine, Shiraz University, Shiraz, Iran Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran School of Veterinary Medicine, Shiraz University, Shiraz, Iran

a r t i c l e

i n f o

Article history: Received 13 May 2014 Received in revised form 9 July 2014 Accepted 13 July 2014 Keywords: Small Indian mongoose (Herpestes Javanicus) Candidatus Mycoplasma turicensis 16S rDNA Phylogenetic analysis

a b s t r a c t Hemoplasmas are the trivial name for a group of erythrocyte-parasitizing bacteria of the genus Mycoplasma. This study is the first report of hemoplasma infection in Small Indian Mongoose (Herpestes Javanicus) based on molecular analysis of 16S rDNA. Whole blood samples were collected by sterile methods, from 14 live captured mongooses, in the south of Iran. Candidatus Mycoplasma turicensis (CMt)-like hemoplasma was detected in blood samples from one animal tested. BLAST search and phylogenetic analysis of partial 16S rDNA sequence (933 bp) of the hemoplasma from Small Indian mongoose (KJ530704) revealed only 96–97% identity to the previously described CMt followed by 95% and 91% similarity with Mycoplasma coccoides and Mycoplasma haemomuris, respectively. Accordingly, the Iranian mongoose CMt isolate showed a high intra-specific genetic variation compared to all previously reported CMt strains in GenBank. Further molecular studies using multiple phylogenetic markers are required to characterize the exact species of Mongoose-derived hemoplasma. © 2014 Elsevier Ltd. All rights reserved.

1. Introduction Hemotropic mycoplasmas (HM), also known as hemoplasmas, a subset of the Mycoplasma genus, are uncultivable, cell wall-free bacteria that attach to the red blood cells (RBCs) of numerous mammalian species such as horses, cattle, cats and dogs [1–3] and can induce lifethreatening hemolytic anemia in infected animals [4,5].

∗ Corresponding author at: Department of Clinical Studies, School of Veterinary Medicine, Shiraz University, Shiraz, P.O. Box 1731, Shiraz 71345, Iran. Tel.: +98 9177023010. E-mail address: nazifi@shirazu.ac.ir (S. Nazifi).

In felids, three important different hemoplasma species have been identified: Mycoplasma haemofelis (Mhf), “Candidatus Mycoplasma haemominutum” (CMhm) and, recently, “Candidatus Mycoplasma turicensis” (CMt) [6–9]. In general, acute infection of cats with either Mhf or CMt is associated with mild to severe anemia, whereas infection with CMhm usually results in few clinical signs [8,10]. Feline hemoplasma species have been detected in an increasing number of mammalian species worldwide, including wild and domestic felids with geographic variation of prevalence [11,12]. However, the molecular epidemiology of these agents and the genetic diversity of their strains worldwide are currently poorly defined, especially in wild animal species.

http://dx.doi.org/10.1016/j.cimid.2014.07.002 0147-9571/© 2014 Elsevier Ltd. All rights reserved.

Please cite this article in press as: Sharifiyazdi H, et al. Molecular characterization and phylogenetic analysis of the causative agent of hemoplasma infection in small Indian Mongoose (Herpestes Javanicus). Comp Immunol Microbiol Infect Dis (2014), http://dx.doi.org/10.1016/j.cimid.2014.07.002

G Model CIMID-970; No. of Pages 5

2

ARTICLE IN PRESS

H. Sharifiyazdi et al. / Comparative Immunology, Microbiology and Infectious Diseases xxx (2014) xxx–xxx

Within each feline hemoplasma species, the 16S rDNA gene sequences from domestic cat-derived isolates of different origin exhibit high sequence identities (>99%) [13,14]. The occurrence of feline hemoplasma infections in wild felids has only marginally been addressed to date [11,15], and may have a role as reservoirs and asymptomatic carriers of hemoplasma infections. Previous molecular analysis of hemoplasma isolates from wild felids i.e., Iberian lynx, Eurasian lynx, European wildcat, lion, puma, oncilla, Geoffroy’s cat, margay, and ocelot showed high sequence identities in the 16S rDNA gene sequence when compared to domestic cat-derived isolates of the same hemoplasma species [11]. The Small Indian Mongoose (Herpestes Javanicus) is native to some parts of Asia mainly India, Iran, Saudi Arabia, and Malaysia. It is a small predatory mammal capable of surviving in a variety of habitats including deserts, forests, agricultural areas and urban areas, often close to human habitation [16]. H. Javanicus represents one of the 10 species of Herpestes. The genus is just one of the genera within the mongoose family Herpestidae. Mongoose are related to civets and cats as part of the sub-order Feliformia, meaning “cat-like” carnivores [16]. But the taxonomy of H. Javanicus is unclear and often debated. Despite having a wide distribution, the parasitic and bacterial infections, especially hemotropic mycoplasmas infection have not been investigated in this host species. Therefore, the aims of the present study were to examine the prevalence of hemoplasma infection in mongoose and characterize the nature of the causative agent(s) by molecular and phylogenetic analyses. 2. Material and methods 2.1. Study area and animals sampling This study was undertaken during a period between June 2012 and July 2013, in an area located at about 15 km north of Shiraz (Badjgah, 29◦ 71 N, 52◦ 59 E), in the south of Iran. A total of 14 mongooses were caught using live trap boxes in most available habitats. Captured animals were anesthetized with ketamine/xylazine during sampling and management programs, then animals were released after markup. To conduct the study, 2 ml Ethylene diamine tetra acetic acid (EDTA)-anticoagulated blood was obtained from the heart. Hematological examination was carried out using automatic cell counter (Exigo, Veterinary Hematology System, Sweden) and blood smears were prepared for Giemsa staining. The remaining blood samples were stored at −20 ◦ C until molecular analysis. 2.2. Molecular analysis: PCR and sequencing Genomic DNA was obtained from 100 ␮l EDTA whole blood samples using a commercial DNA extraction kit (Qiagen DNAeasy; Valencia, California) according to the manufacturer’s protocol. The purity and concentration of the DNA were estimated by spectrophotometry at 260 and 280 nm and stored at −20 ◦ C for subsequent PCR assay.

To screen specimens for HM a PCR assay able to detect the entire group of known HM was performed according to Hoelzle et al. [17]. Regions of 16S rDNA gene (975 bp) were amplified with the forward primer 16S HAEMOforw: 5 -GGCCCATATTCCTRCGGGAAG-3 and the reverse primer 16S HAEMOrev: 5 -ACRGGATTACTAGTGATTCCA-3 using the lyophilized PCR micro tubes (Accupower PCR PreMix, BioNeer Co., Korea). PCR was performed in a thermocycler with the following program: An initial denaturation step at 95 ◦ C for 5 min, 35 cycles of denaturation at 94 ◦ C for 1 min, annealing at 55 ◦ C for 1 min, and extension at 72 ◦ C for 1 min, followed by a final extension at 72 ◦ C for 5 min. DNA samples from known 16S rDNA hemoplasma positive case (KC762746) and water sample were used as positive and negative controls, respectively. The amplified products were purified with a PCR product purification kit (BioNeer, Korea) and sequenced directly using the capillary DNA analyzer (ABI 3730; Applied Biosystems, Foster City, CA, USA) after sequencing reactions with a BigDye Terminator V3.1 cycle sequencing Kit (Applied Biosystems). Forward and reverse nucleic acid sequence data were used to construct a continuous sequence of each inserted DNA. The 16S rDNA sequence obtained was compared to GenBank entries using the BLAST tool provided by NCBI (http://www.ncbi.nlm.nih.gov/blast/Blast.cgi). 2.3. Phylogenetic analyses To classify the sequences derived from mongoose blood the nucleotide sequence data was separately aligned against homologous sequences existing in GenBank by MEGA 4.0. Creating multiple-sequence alignment was established using Clustal W program in the MEGA 4.0 software for each query DNA sequence. Data sequences were also used for construction of the phylogenetic trees using maximum parsimony and neighbor-jointed methods. To assess the robustness of the branches, a bootstrap test with 1000 replicates was performed, following the rule of branch consistency [18]. 3. Results Among all mongooses analyzed for hemoplasma species, only one sample (7.14%) was positive for HM using a PCR assay. Fragment of the positive sample (975 bp) was sequenced, analyzed, and compared with known corresponding sequences in the GenBank database. Chromatogram obtained from sequencing of the PCR product showed a high quality. Our sequence was designated “Iranian Candidatus Mycoplasma turicensis-like hemoplasma” and submitted to the Genbank database with the accession number KJ530704. Comparative sequence analysis using obtained 16S rDNA sequence (KJ530704: 933 bp) demonstrated the highest homology (96–97%) to CMt, previously described in cat, wildcat, Ocelot, Iriomote cat and lion. However, sequence analyses of the 16S rDNA gene identified a distinct genotype for Iranian mongoose isolates compared with those of sequences for other CMt deposited in the GenBank database. When the nucleotide sequence of the partial

Please cite this article in press as: Sharifiyazdi H, et al. Molecular characterization and phylogenetic analysis of the causative agent of hemoplasma infection in small Indian Mongoose (Herpestes Javanicus). Comp Immunol Microbiol Infect Dis (2014), http://dx.doi.org/10.1016/j.cimid.2014.07.002

G Model CIMID-970; No. of Pages 5

ARTICLE IN PRESS

H. Sharifiyazdi et al. / Comparative Immunology, Microbiology and Infectious Diseases xxx (2014) xxx–xxx

3

16S rDNA gene was compared with those of Mycoplasma coccoides and Mycoplasma haemomuris our hemoplasma shared 95% (883/933 bp) homology with M. coccoides and 91% (852/933) with M. haemomuris. Interestingly, all previous CMt derived 16S rDNA gene sequences showed a lower identity to those of M. coccoides (93–94%). Furthermore, phylogenetic analysis of concatenated data showed all previous CMt hemoplasma species were located within a single major group based on the phylogenetic tree whereas our CMt-like hemoplasma isolate branched away from other CMt and was located in a separated sub-group between CMt group and a rodent hemoplasma species (M. coccoides) (Fig. 1). In accordance with the phylogenetic results which mentioned Iranian mongoose, CMt-like hemoplasma isolate showed a noticeable intra-specific diversity (3.1–4.2%) compared to other existing CMt isolates in the GenBank. Previous level of intra-specific 16S rDNA variation existed in CMt isolates worldwide ranged from 0% to 2.4%. The bootstrap percentage values are given at the nodes of the phylogenetic tree shown in Fig. 1. Hematological results revealed that the infected mongoose did not exhibit any significant alterations in hematological values (hemoglobin (Hb), hematocrit (HCT) and red blood cells (RBCs)) in comparison to hemoplasma PCR-negative animals. Abnormalities noted in the stained blood smear for the positive case compared to other negative animals included mild increases in Howell–Jolly bodies and nucleated RBCs. 4. Discussion In recent years, there has been growing interest in hemotropic mycoplasmas, the causative agents of infectious anemia in a wide range of mammalian species [19]. These hemotropic bacteria were previously classified in the order Rickettsiales as a member of the family Anaplasmataceae. However, sequence analysis of the 16S rRNA gene showed that these bacteria were members of the genus Mycoplasma [19–21]. The development of molecular assays, which target primarily the 16S rDNA of these cell wall-deficient uncultivable microbes, has resulted in the recent recognition of several novel animal hemoplasmas [22,23]. The genetic diversity of these pathogens worldwide is currently poorly defined. Hemoplasmas are obligate epierythrocytic organisms that attach to erythrocytes, appear to be relatively nonpathogenic, and are visualized on blood smears more often during periods of stress, hard work, or concurrent infection [23]. In some animals, hemoplasma infection is associated with hemolytic anemia of variable severity, ranging from nonclinical hemolysis to severe anemia [11]. This study provides the first molecular evidence of a CMt-like hemoplasma infection in a mongoose. However, the prevalence of hemoplasma infection in this host species seems to be low (7.14%). The lack of clear clinical signs in the infected mongoose could be explained by the possibility that this case was sampled during a chronic carrier status and not during acute infection. Peripheral blood smear of infected animal showed only a

Fig. 1. Bootstrap phylogenetic analysis of the partial 16S rDNA gene sequences of our mongoose isolate Mycoplasma species (KJ530704) and related organisms. The phylogenetic tree was constructed using the neighbor-joining method, and evolutionary distances are to the scale shown. Bootstrap percentage values are given at the nodes of the tree.

slight increase in Howell–Jolly bodies and nucleated red blood cells (NRBC). However, hemoplasma PCR-positive mongoose did not exhibit significantly lower hemoglobin (Hb), hematocrit (HCT) and red blood cells (RBCs) values than did hemoplasma PCR-negative animals. Accordingly, our results suggest that hemoplasma infections might not pose a serious threat to the Small Iranian Indian mongoose population. Similarly, previous researchers reported

Please cite this article in press as: Sharifiyazdi H, et al. Molecular characterization and phylogenetic analysis of the causative agent of hemoplasma infection in small Indian Mongoose (Herpestes Javanicus). Comp Immunol Microbiol Infect Dis (2014), http://dx.doi.org/10.1016/j.cimid.2014.07.002

G Model CIMID-970; No. of Pages 5

4

ARTICLE IN PRESS

H. Sharifiyazdi et al. / Comparative Immunology, Microbiology and Infectious Diseases xxx (2014) xxx–xxx

that domestic cats recovering from acute infection usually lack signs of anemia although they test PCR positive and occasionally show high hemoplasma loads [9,10,24]. In addition, Willi et al. [11] also reported a clinically healthy Iberian lynx in their study that remained PCR positive for “CMhm” over a 22-month follow-up period. Phylogenetic analysis of the 16S rDNA gene of Iranian mongoose isolate showed this parasite belonged to the haemofelis cluster that included CMt, M. coccoides, M. haemomuris, Mycoplasma haemobos, M. haemofelis and Mycoplasma haemocanis. The hemotropic parasite infecting the mongoose was, unexpectedly, quite distinct from the sequences of 27 other CMt strains previously published, with ≤97% identities only. Phylogenetic analysis of the isolate showed it belonged to the CMt group and forms a sister group to the M. coccoides, which is rodent hemotropic mycoplasmal species. Phylogenetic tree suggested that CMt and M. coccoides may have been generated from a single common ancestor. The close relationship between the CMt and rodent hemoplasmas suggest potential interspecies transmission of these agents [8,22]. M. coccoides has been shown to be mechanically transmitted between mice through the mouse louse Polyplax serrata [25]. Willi et al. [12] mentioned that an interspecies transmission of hemoplasma from mouse to cat could have taken place through hunting. Furthermore, bloodsucking arthropods may play a role in the transmission of hemoplasmas in different host species worldwide [11]. As suggested by some authors, a relative important zoonotic role of such pathogen should be considered. Sporadic reports described organisms with morphological similarities to hemoplasmas in the blood of human patients [26,27]. Furthermore, a recently published report demonstrated the first molecular detection of a feline hemoplasma (M. haemofelis) species in an immunocompromised HIV-positive human patient in Brazil that supports the hypothesis of the zoonotic potential of these agents [28]. Maggi et al. [29] mentioned that the pathogenic potential of many hemotropic Mycoplasmas as a cause of human and animal disease remains to be elucidated as do the mechanisms of intra- and inter-species transmission. Because of the lifestyle mongooses that can live in almost any environment where cats and rodents are present and according to the principal modes of transmission for infection between animals, i.e., insect bites: lice, fleas, mosquitoes, midges and stable flies, this new host species must be considered as a potential reservoir for CMt. CMt infection has been reported in domestic cats in Switzerland [9,23], the United States [30], the United Kingdom, Australia, and South Africa [14] and in wild felid species from Spain, France, Switzerland, Tasmania, and Brazil [11] and Phylogenetic analyses revealed that most CMt isolates from wild felids were indeed very closely related to CMt from domestic cats [11]. In contrast, the CMt-like hemoplasma isolate from mongoose branched away from domestic cat isolates and could not be completely assigned to CMt subcluster and formed different subclusters between CMt group and M. coccoides in the phylogenetic tree (Fig. 1). This finding is surprising because most isolates from felids from all over the world could

be clearly assigned to the CMt subcluster. It has been speculated that CMt strains from different clusters differ in pathogenicity, geographical distribution or host origin. However, in addition to the gene of interest, numerous other genes must be simultaneously examined to clarify the correlation between phylogenetic lineage and these characteristics. In conclusion, we found that CMt-like hemoplasma is present in mongoose population in Iran but the prevalence of this hemoplasma infection seems relatively low. Furthermore, the Iranian mongoose CMt-like hemoplasma isolate showed a high intra-specific genetic variation compared to all previous CMt isolates. However, further studies using a large-scale molecular phylogenetic analysis are required to clarify the epidemiological importance and role of the present hemoplasma genotype of CMt in domestic and wild animals’ hemoplasma infection, especially in Iran. Conflict of interest We declare that we have no conflict of interest. Acknowledgments The researchers would like to thank the Research Council of Shiraz University and School of Veterinary Medicine, Shiraz University for financial and technical support of this study (Grant No. 71-GR-VT-5). The authors would specifically like to thank Mr. Rahsepar and Mr. Hosseini for their laboratory assistance. References [1] Hoelzle LE. Hemotrophic mycoplasmas: recent advances in Mycoplasma suis. Vet Microbiol 2008;130:215–26. [2] Groebel K, Hoelzle K, Wittenbrink MM, Ziegler U, Hoelzle LE. Mycoplasma suis invades porcine erythrocytes. Infect Immun 2009;77:576–84. [3] Dieckmann SM, Winkler M, Groebel K, Dieckmann MP, HofmannLehmann R, Hoelzle K, et al. Haemotrophic mycoplasma infection in horses. Vet Microbiol 2010;145:351–3. [4] Messick JB. New perspectives about Hemotrophic mycoplasma (formerly, Haemobartonella and Eperythrozoon species) infections in dogs and cats. Vet Clin North Am: Small Anim Pract 2003;33:1453–65. [5] Messick JB. Hemotrophic mycoplasmas (hemoplasmas): a review and new insights into pathogenic potential. Vet Clin Pathol 2004;33:2–13. [6] Foley JE, Pedersen NC. ‘Candidatus Mycoplasma haemominutum’, a low-virulence epierythrocytic parasite of cats. Int J Syst Evol Microbiol 2001;51:815–7. [7] Neimark H, Johansson KE, Rikihisa Y, Tully JG. Proposal to transfer some members of the genera Haemobartonella and Eperythrozoon to the genus Mycoplasma with descriptions of ‘Candidatus Mycoplasma haemofelis’, ‘Candidatus Mycoplasma haemomuris’, ‘Candidatus Mycoplasma haemosuis’ and ‘Candidatus Mycoplasma wenyonii’. Int J Syst Evol Microbiol 2001;51:891–9. [8] Willi B, Boretti FS, Cattori V, Tasker S, Meli ML, Reusch C, et al. Identification, molecular characterization, and experimental transmission of a new hemoplasma isolate from a cat with hemolytic anemia in Switzerland. J Clin Microbiol 2005;43:2581–5. [9] Willi B, Boretti FS, Baumgartner C, Tasker S, Wenger B, Cattori V, et al. Prevalence, risk factor analysis, and follow-up of infections caused by three feline hemoplasma species in cats in Switzerland. J Clin Microbiol 2006;44:961–9. [10] Foley JE, Harrus S, Poland A, Chomel B, Pedersen NC. Molecular, clinical, and pathologic comparison of two distinct strains of Haemobartonella felis in domestic cats. Am J Vet Res 1998;59:1581–8.

Please cite this article in press as: Sharifiyazdi H, et al. Molecular characterization and phylogenetic analysis of the causative agent of hemoplasma infection in small Indian Mongoose (Herpestes Javanicus). Comp Immunol Microbiol Infect Dis (2014), http://dx.doi.org/10.1016/j.cimid.2014.07.002

G Model CIMID-970; No. of Pages 5

ARTICLE IN PRESS

H. Sharifiyazdi et al. / Comparative Immunology, Microbiology and Infectious Diseases xxx (2014) xxx–xxx [11] Willi B, Filoni C, Catao-Dias JL, Cattori V, Meli ML, Vargas A, et al. Worldwide occurrence of feline hemoplasma infections in wild felid species. J Clin Microbiol 2007;45:1159–66. [12] Willi B, Novacco M, Meli M, Wolf-Jackel G, Boretti F, Wengi N, et al. Haemotropic mycoplasmas of cats and dogs: transmission, diagnosis, prevalence and importance in Europe. Schweiz Arch Tierheilkd 2010;152:237–44. [13] Tasker S, Helps CR, Day MJ, Harbour DA, Shaw SE, Harrus S, et al. Phylogenetic analysis of hemoplasma species: an international study. J Clin Microbiol 2003;41:3877–80. [14] Willi B, Tasker S, Boretti FS, Doherrm MG, Cattori V, Meli ML, et al. Phylogenetic analysis of “Candidatus Mycoplasma turicensis” isolates from pet cats in the United Kingdom, Australia, and South Africa, with analysis of risk factors for infection. J Clin Microbiol 2006;44:4430–5. [15] Haefner M, Burke TJ, Kitchell BE, Lamont LA, Schaeffer DJ, Behr M, et al. Identification of Haemobartonella felis (Mycoplasma haemofelis) in captive nondomestic cats. J Zoo Wildl Med 2003;34:139–43. [16] Csurhes S, Fisher P. Pest animal risk assessment: Indian mongoose (Herpestes javanicus). Queensland Government Publications; 2010. p. 1–13. [17] Hoelzle K, Winkler M, Kramer MM, Wittenbrink MM, Dieckmann SM, Hoelzle LE. Detection of Candidatus Mycoplasma haemobos in cattle herds with anaemia in Germany. Vet J 2010;187(3):408–10. [18] Tamura K, Dudley J, Nei M, Kumar S. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol 2007;24:1596–9. [19] Willi B, Boretti FS, Tasker S, Meli ML, Wengi N, Reusch CE, et al. From Haemobartonella to hemoplasma: molecular methods provide new insights. Vet Microbiol 2007;125:197–209. [20] Neimark H, Kocan KM. The cell wall-less rickettsia Eperythrozoon wenyonii is a mycoplasma. FEMS Microbiol Lett 1997;156:287–91. [21] Rikihisa Y, Kawahara M, Wenyon B, Kociba G, Fuerst P, Kawamori F, et al. Western immunoblot analysis of Haemobartonella muris and

[22]

[23]

[24]

[25] [26]

[27]

[28]

[29]

[30]

5

comparison of 16S rRNA gene sequences of H. muris, H. felis, and Eperythrozoon suis. J Clin Microbiol 1997;35:823–9. Sykes JE, Bailiff NL, Ball LM, Foreman O, George JW, Fry MM. Identification of a novel hemotropic mycoplasma in a splenectomized dog with hemic neoplasia. J Am Vet Med Assoc 2004;224: 1946–51. Willi B, Boretti FS, Baumgartner C, Cattori V, Meli ML, Doherr MG, et al. Feline hemoplasmas in Switzerland: identification of a novel species, diagnosis, prevalence, and clinical importance. Schweiz Arc Tierheilkd 2006;148:139–40. Tasker S, Caney SM, Day MJ, Dean RS, Helps CR, Knowles TG, et al. Effect of chronic FIV infection, and efficacy of marbofloxacin treatment, on Mycoplasma haemofelis infection. Vet Microbiol 2006;117:169–79. Berkenkamp SD, Wescott RB. Arthropod transmission of Eperythrozoon coccoides in mice. Lab Anim Sci 1988;38:398–401. Maggi RG, Compton SM, Trull CL, Mascarelli PE, Mozayeni BR, Breitschwerdt EB. Infection with hemotropic Mycoplasma species in patients with or without extensive arthropod or animal contact. J Clin Microbiol 2013;51(10):3237–41. Steer JA, Tasker S, Barker EN, Jensen J, Mitchell J, Stocki T, et al. A novel hemotropic Mycoplasma (hemoplasma) in a patient with hemolytic anemia and pyrexia. Clin Infect Dis 2011;53(11):147–51. dos Santos AP, dos Santos RP, Biondo AW, Dora JM, Goldani LZ, de Oliveira ST, et al. Hemoplasma infection in HIV positive patient, Brazil. Emerg Infect Dis 2008;14:1922–4. Maggi RG, Chitwood MC, Kennedy-Stoskopf S, DePerno CS. Novel hemotropic Mycoplasma species in white-tailed deer (Odocoileus virginianus). Comp Immunol Microbiol Infect Dis 2013;36(6): 607–11. Sykes JE, Terry JC, Lindsay LL, Owens SD. Prevalences of various hemoplasma species among cats in the United States with possible hemoplasmosis. J Am Vet Med Assoc 2008;232:372–9.

Please cite this article in press as: Sharifiyazdi H, et al. Molecular characterization and phylogenetic analysis of the causative agent of hemoplasma infection in small Indian Mongoose (Herpestes Javanicus). Comp Immunol Microbiol Infect Dis (2014), http://dx.doi.org/10.1016/j.cimid.2014.07.002