Ovine papillomaviruses: Diversity, pathogenicity, and evolution.

The family Papillomaviridae includes a plethora of viral species infecting virtually all vertebrates excluding amphibians, with astonishing impact on human and animal health. Although more than 250 species have been described in humans, the total number of papillomaviruses (PVs) discovered in animals does not reach up to this number. In animals, PV infections are mostly asymptomatic or can cause variable clinical conditions ranging from self-limiting papillomas and other cutaneous and mucosal benign lesions to cancer. Most of animal PV types have been discovered in cattle, dogs, horses, and cats with other farm host species remaining overlooked. In particular, the number of PV types so far identified in sheep is limited. This paper comprehensively reviews ovine PVs features, including viral taxonomy and evolution; genome organization; viral tropism and pathogenesis; macroscopical features and histopathological patterns, as well as available diagnostics tools. Data are critically presented and discussed in terms of impact on veterinary and public health. The development of future dedicated research is also discussed.

Molecular typing of bovine papillomaviruses in Costa Rica.

Bovine papillomaviruses are related to cause fibroepithelial proliferations in the skin and mucosae and are associated with economic loss mainly related to poor body condition and reduced milk production. This study aimed to investigate the presence and types of bovine papillomaviruses (BPVs) in cattle sampled in different areas of Costa Rica using molecular techniques. A descriptive study with a non-probability convenience sampling was carried out. A total of 99 papillomatous lesions were collected from 63 animals in 32 farms, and analyzed by polymerase chain reaction, rolling circle amplification (RCA), sequencing, and restriction enzymes digestion. Seven bovine papillomavirus types (BPV1, BPV2, BPV4, BPV6, BPV7, BPV10, BPV11) and two putative novel viral variants (BPV-CR1 and BPV-CR2) were identified for the first time in Costa Rica. BPV6 was the most frequently detected virus in lesions (31.2%), followed by BPV2 (25%) and BPV1 (25%). BPV1 and BPV2 were the most widely distributed in the Country. Coinfections were recorded in two animals (BPV1 / BPV2 and BPV4 / BPV6). Restriction analyses allowed differentiating BPV1 from BPV2, BPV4, and BPV7, but failed to identify BPV6, BPV10, and BPV11. Results suggest that a great PVs diversity is harbored by bovines in Costa Rica and indicate the need for further investigations aimed to uncover PV diversity at the full genomic level.

First Molecular Detection of Zoonotic Chlamydia Species in Vietnamese Goats.

The genus Chlamydia comprises obligate intracellular bacteria that infect a wide variety of hosts, with infection leading to a range of diseases in humans and animals; they thus constitute a major public health threat. Among the members of the Chlamydiaceae family, Chlamydia suis, C. abortus, C. pecorum, and C. psittaci represent the most important pathogenic species infecting a large range of hosts and are a well-established threat to livestock. Information regarding the circulation of Chlamydia species in ruminants from Vietnam is lacking. In this study, DNA extracted from 60 blood samples collected from goats in Hue province was used for Chlamydia spp. identification by classic PCR and Sanger sequencing. Chlamydia spp. were detected in eleven samples (18.3%) and C. abortus and C. psittaci were molecularly identified by sequencing. Despite the limited sample size in this study, findings point out the relevance of ruminants as hosts of chlamydial species in Central Vietnam and the importance of monitoring chlamydial strains through the activation of surveillance programs in this country. The need for a deeper evaluation of human and animal health risk analysis in terms of chlamydiosis should be also considered.

Genome typing, histopathology, and evolution of BPV30, a novel Xipapillomavirus type isolated from Bovine papilloma in Costa Rica.

Xipapillomavirus includes a group of viruses almost exclusively reported in both beef cattle and dairy breeding, in which they induce papillomatosis and occasionally malignant tumors. Bovine papillomaviruses (BPVs) infection impacts greatly on animal productions, and this is amplified by their cosmopolitan distribution. Cutaneous proliferative lesions in bovines can relate to leather depreciation and impaired milk production by giving rise to obstruction of the teat and hygiene limitations, often leading to hemorrhagic mastitis. This study reports the identification of a novel Xipapillomavirus type associated with udder papilloma in a Jersey cow in Costa Rica. Viral genome was fully sequenced and molecularly characterized. Histopathology and viral phylogeny and evolution are also presented and discussed by comparison with already described BPVs. Based on results, a novel Xipapillomavirus type, namely BPV30, is proposed. BPV30 is a typical Xipapillomavirus 2 most similar to BPV12, from which it separated roughly 18 million years ago. The absence of E6 and the presence of E10 in BPV30 confirm an E6 loss occurring along the clade leading to BPV12. The identification of this novel BPV is fundamental to the development of specific prophylactic tools, which represent the most effective weapon to fight viral circulation, to prevent infections, and eventually controlling associated proliferative lesions.

Emergence of Anaplasma Species Related to A. phagocytophilum and A. platys in Senegal.

The genus Anaplasma (Anaplasmataceae, Rickettsiales) includes tick-transmitted bacterial species of importance to both veterinary and human medicine. Apart from the traditionally recognized six Anaplasma species (A. phagocytophilum, A. platys, A. bovis, A. ovis, A. centrale, A. marginale), novel strains and candidate species, also of relevance to veterinary and human medicine, are emerging worldwide. Although species related to the zoonotic A. platys and A. phagocytophilum have been reported in several African and European Mediterranean countries, data on the presence of these species in sub-Saharan countries are still lacking. This manuscript reports the investigation of Anaplasma strains related to zoonotic species in ruminants in Senegal by combining different molecular tests and phylogenetic approaches. The results demonstrated a recent introduction of Candidatus (Ca) Anaplasma turritanum, a species related to the pathogenic A. platys, possibly originating by founder effect. Further, novel undetected strains related to Candidatus (Ca) Anaplasma cinensis were detected in cattle. Based on groEL and gltA molecular comparisons, we propose including these latter strains into the Candidatus (Ca) Anaplasma africanum species. Finally, we also report the emergence of Candidatus (Ca) A. boleense in Senegal. Collectively, results confirm that Anaplasma species diversity is greater than expected and should be further investigated, and that Anaplasma routine diagnostic procedures and epidemiological surveillance should take into account specificity issues raised by the presence of these novel strains, suggesting the use of a One Health approach for the management of Anaplasmataceae in sub-Saharan Africa.

gltA typing of Anaplasma strains related to A. platys: Taxonomical and one health implications.

Species belonging to the genus Anaplasma (Rickettsiales) include bacteria of veterinary and public health importance. Beside the zoonotic Anaplasma phagocytophilum, A. platys, the etiological agent of canine cyclic thrombocytopenia, has been sporadically reported in clinically ill human patients. The ongoing emergence of novel strains related to this species in vertebrate hosts emphasises the need for genetic comparisons among strains identified in different regions of the world. In this paper we developed a PCR test suitable for amplification of the still undescribed gltA gene of Anaplasma strains related to A. platys from Mediterranean ruminants and applied on a panel of 248 samples. gltA sequencing allowed phylogenetic comparison with strains related to A. platys recently identified in China, and strains representative of the Anaplasmataceae family. Results suggest the designation of Candidatus A. turritanum, including Mediterranean A. platys - like strains, and Candidatus A. cinensis, including strains isolated in China. Data generated in this study are a solid reference for future epidemiological studies of novel unclassified strains related to A. platys and for their diagnosis and raise concern on their potential veterinary and public health implications encouraging investigating the suspected unexplored diversity within the genus Anaplasma in animals and human.

Molecular survey of parvovirus, astrovirus, coronavirus, and calicivirus in symptomatic dogs.

Gastrointestinal disorders caused by enteric viruses are frequently reported in dogs worldwide, with significant mortality rates in unvaccinated individuals. This study reports the identification and molecular characterization of Canine parvovirus (CPV-2), Canine coronavirus (CcoV), Canine astrovirus (AstV), and Canine calicivirus (CcaV) in a panel of dogs showing severe enteric clinical signs sampled in a typical Mediterranean environment (Sardinia, Italy). At least one of these viral species was detected in 92.3% samples. CPV-2 was the most frequently detected virus (87.2%), followed by AsTv (20.5%), CCoV-IIa (18%), and CCoV-I (10.3%). CCoV-IIb and CaCV were not detected in any sample. Single infection was detected in 24 samples (66.7%), mainly related to CPV-2 (91.7%). Coinfections were present in 33.3% samples with constant detection of CPV-2. Canine coronavirus was present only in coinfected animals. The VP2 sequence analysis of CPV-2 positive samples confirmed the presence of all variants, with CPV-2b most frequently detected. Phylogeny based on the CcoV-IIa spike protein (S) gene allowed to identify 2 different clades among Sardinian isolates but failed to distinguish enteric from pantropic viruses. Study on presence and prevalence of enteroviruses in dogs increase our knowledge about the circulation of these pathogens in the Mediterranean area and highlight the need for dedicated routine vaccine prophylaxis. Molecular analyses of enteric viruses are fundamental to avoid failure of vaccines caused by frequent mutations observed in these enteroviruses.

Molecular characterization and phylogenetic analysis of Babesia and Theileria spp. in Sardinian ruminants.

Tick-borne diseases (TBDs) caused by Theileria and Babesia spp. are common in tropical and subtropical regions. This study investigates the presence of Theileria and Babesia spp. in ruminants from a subtropical Mediterranean region (Sardinia, Italy), a hotspot for ticks infestations. A total of 141 blood samples from healthy and symptomatic ruminants (showing symptoms consistent with tick-borne disease) were screened using a polymerase chain reaction test based on the amplification of the 18 s rRNA fragment. A total of 19/50 sheep (38%), 34/43 bovine (79.1%), and 5/48 goats (10.4%) tested positive to Babesia/Theileria. Phylogenetic analysis assigned all sequences obtained from sheep to the T. ovis cluster, while bovine and goats sequence types grouped in the Theileria buffeli/sergenti/orientalis group. One sequence type, isolated from a symptomatic bovine, clustered with B. major. Information on presence and frequency of piroplasms in ruminants increase our knowledge about the circulation of these pathogens in Sardinian animals and add up to previous studies conducted in ticks in the same area. Results also highlight the importance of subtropical Mediterranean environments as hotspots for ruminants piroplasmosis with potential impact on Veterinary Health.

Histopathological and Molecular Study of Pacific Oyster Tissues Provides Insights into V. aestuarianus Infection Related to Oyster Mortality.

Consumer preference for healthy and sustainable food products has been steadily increasing in recent years. Bivalve mollusks satisfy these characteristics and have captured ever-increasing market shares. However, the expansion of molluscan culture in worldwide and global trade have favored the spread of pathogens around the world. Combined with environmental changes and intensive production systems this has contributed to the occurrence of mass mortality episodes, thus posing a threat to the production of different species, including the Pacific oyster Crassotrea gigas. In the San Teodoro lagoon, one of the most devoted lagoons to extensive Pacific oyster aquaculture in Sardinia, a mortality outbreak was observed with an estimated 80% final loss of animal production. A study combining cultural, biomolecular and histopathological methods was conducted: (1) to investigate the presence of different Vibrio species and OsHV-1 in selected oyster tissues (digestive gland, gills, and mantle); (2) to quantify Vibrio aestuarianus and to evaluate the severity of hemocyte infiltration in infected tissues; (3) to produce post-amplification data and evaluating ToxR gene as a target for phylogenetic analyses. Results provide new insights into V. aestuarianus infection related to oyster mortality outbreaks and pave the way to the development of tools for oyster management.

Genomic characterization of a novel bat-associated Circovirus detected in European Miniopterus schreibersii bats.

Circoviruses are small circular DNA viruses causing severe pig and poultry disease, recently identified in various bat species worldwide. We report the detection and full-genome molecular characterization of a novel bat-associated Circovirus identified in faecal samples of Miniopterus schreibersii bats (Schreiber's bent-winged bats) from Sardinia, Italy. Full-genomic sequencing revealed a new putative member of Circoviridae family, with a genome size of 2063 nt. Sequencing allowed the characterization of the two major ORFs, inversely arranged, encoding replicase and capsid proteins, as well as the finding of a polythymidine tract within the genome, and highlighted phylogenetic relationships of the novel virus. This is the first report of circovirus in European bats. Giving the high level of genetic diversity of bat circoviruses, it is paramount to further investigate the relationships between these viruses and bats.

Leopardus wiedii Papillomavirus type 1, a novel papillomavirus species in the tree ocelot, suggests Felidae Lambdapapillomavirus polyphyletic origin and host-independent evolution.

The limited knowledge on Papillomavirus diversity (particularly in wild animal species) influences the accuracy of PVs phylogeny and their evolutionary history, and hinders the comprehension of PVs pathogenicity, especially the mechanism of virus - related cancer progression. This study reports the identification of Leopardus wiedii Papillomavirus type 1 (LwiePV1), the first PV type within Lambdapapillomavirus in a Leopardus host. LwiePV1 full genome sequencing allowed the investigation of its taxonomic position and phylogeny. Based on results, LwiePV1 should be assigned to a novel PV species providing evidence for a polyphyletic origin of feline lambda PVs, and representing an exception to codivergence between feline lambda PVs and their hosts. Results improve our knowledge on PV diversity and pave the way to future studies investigating biological and evolutionary features of animal PVs.

Molecular epidemiology of Anaplasma spp. related to A. phagocytophilum in Mediterranean small ruminants.

The genus Anaplasma currently comprises 6 bacterial species mostly pathogenic to animals and/or human, including the zoonotic species Anaplasma phagocytophilum, the causative agent of tick-borne fever (TBF) of ruminants, and of granulocytic anaplasmosis of horses, dogs and human. Recently, novel potentially non-pathogenic strains related to A. phagocytophilum have been identified in Japan, China, and Tunisia. This paper reports the identification, molecular typing, and evolutionary history of novel Anaplasma strains (A. phagocytophilum-like 1 and 2), related to but distinct from A. phagocytophilum in Mediterranean area of Europe and Africa. PCR-RFLP and phylogenetic analyses based on 16S rRNA provided evidence for the circulation of A. phagocytophilum-like 1 strains in Europe. Phylogeny based on groEL gene showed the inclusion of Sardinian and Tunisian A. phagocytophilum-like 1 strains in a unique clade distinct from, but related to that of Japanese strains. Results suggest that genetic diversity within the genus Anaplasma is much greater than expected and provide information useful for the development of specific and effective diagnostic and prophylactic tools.

Molecular characterization and phylogenetic analysis of Babesia and Theileria spp. in ticks from domestic and wild hosts in Sardinia.

Piroplasmoses are tick-borne protozoan diseases caused by hemoprotozoan parasites with considerable economic, veterinary and medical impact worldwide. Here, the presence and diversity of piroplasmids was investigated in ticks collected from domestic and wild hosts in a typical subtropical environment of Sardinia island by standard PCR, sequencing, and phylogenetic analyses. We demonstrate the presence of strains closely related to the Theileria buffeli/sergentii/orientalis complex in Rhipicephalus sanguineus s.l., Rh. bursa, Rh. annulatus, Hyalomma marginatum, Dermacentor marginatus and Haemaphysalis punctata ticks. A strain detected in two Rh. sanguineus s.l. ticks collected from dogs grouped with T. equi, the agent of equine piroplasmosis. T. ovis, the main etiological agent of ovine theileriosis, was detected in one Rh. bursa tick from a mouflon. Babesia bigemina, the causative agent of bovine babesiosis, was detected in two Rh. sanguineus s.l. ticks from dogs. Our findings expand the knowledge on the repertoire of tick-borne pathogens present in Mediterranean ticks. Further analyses are needed to determine the role of ticks in the biological or mechanical transmission of piroplasmoses in this area.

Use of 2-dimensional speckle-tracking echocardiography to assess left ventricular systolic function in dogs with systemic inflammatory response syndrome.

BACKGROUND: Early identification of systolic dysfunction in dogs with systemic inflammatory response syndrome (SIRS) potentially could improve the outcome and decrease mortality. OBJECTIVE: To compare 2-dimensional speckle tracking (2D-STE) with 2-dimensional (2D) and M-mode echocardiography in the evaluation of systolic function in SIRS dogs. ANIMALS: Seventeen SIRS and 17 healthy dogs. METHODS: Prospective observational case-control study. Each dog underwent physical examination, conventional echocardiography, 2D-STE, and C-reactive protein measurement. RESULTS: Dogs with SIRS had lower 2D-STE ejection fraction (X4D-EF; 44 ± 8 versus 53 ± 8; P = .003), endocardial global longitudinal strain (ENDO-G-Long-St; -14.6 ± 3.2 versus -18.5 ± 4.1; P = .003), and normalized left ventricular diameter in diastole (1.38 ± 0.25 versus 1.54 ± 0.17; P = .04) and systole (0.85 ± 0.18 versus 0.97 ± 0.11; P = .03) as compared to healthy dogs. Simpson method of disks (SMOD) right parasternal EF (55 ± 9 versus 60 ± 6; P = .07) and end systolic volume index (ESVI; 23 ± 10 versus 21 ± 6; P = .61), SMOD left apical EF (59 ± 9 versus 59 ± 6; P = .87) and ESVI (20 ± 8 versus 22 ± 6; P = .25), fractional shortening (FS; 34 ± 5 versus 33 ± 4; P = .39), M-mode EF (64 ± 7 versus 62 ± 5; P = .35), and ESVI (23 ± 11 versus 30 ± 9; P = .06) were not significantly different between SIRS and control group, respectively. CONCLUSION AND CLINICAL IMPORTANCE: Speckle tracking X4D-EF and ENDO-G-Long-St are more sensitive than 2D and M-Mode FS, EF, and ESVI in detecting systolic impairment in dogs with SIRS.

Molecular analysis of carnivore Protoparvovirus detected in white blood cells of naturally infected cats.

BACKGROUND: Cats are susceptible to feline panleukopenia virus (FPV) and canine parvovirus (CPV) variants 2a, 2b and 2c. Detection of FPV and CPV variants in apparently healthy cats and their persistence in white blood cells (WBC) and other tissues when neutralising antibodies are simultaneously present, suggest that parvovirus may persist long-term in the tissues of cats post-infection without causing clinical signs. The aim of this study was to screen a population of 54 cats from Sardinia (Italy) for the presence of both FPV and CPV DNA within buffy coat samples using polymerase chain reaction (PCR). The DNA viral load, genetic diversity, phylogeny and antibody titres against parvoviruses were investigated in the positive cats. RESULTS: Carnivore protoparvovirus 1 DNA was detected in nine cats (16.7%). Viral DNA was reassembled to FPV in four cats and to CPV (CPV-2b and 2c) in four cats; one subject showed an unusually high genetic complexity with mixed infection involving FPV and CPV-2c. Antibodies against parvovirus were detected in all subjects which tested positive to DNA parvoviruses. CONCLUSIONS: The identification of FPV and CPV DNA in the WBC of asymptomatic cats, despite the presence of specific antibodies against parvoviruses, and the high genetic heterogeneity detected in one sample, confirmed the relevant epidemiological role of cats in parvovirus infection.

GroEL typing and phylogeny of Anaplasma species in ticks from domestic and wild vertebrates.

Anaplasma species are globally distributed tick-borne bacteria causing a range of clinical conditions in domestic animals, wildlife, and human. Nevertheless, data on presence and distribution of Anaplasma strains in ticks are still lacking, especially in the Mediterranean region. This study reports the molecular identification, genetic characterization and phylogeny of Anaplasma strains of both veterinary and zoonotic importance in ticks collected from domestic and wild hosts sampled in a typical Mediterranean warm temperate region, the island of Sardinia. Results reveal the presence of A. ovis in Rhipicephalus bursa and R. sanguineus sensu lato ticks; A. platys-like was found in R. bursa ticks; while A. platys and A. phagocytophilum were detected in Hyalomma marginatum and H. lusitanicum ticks. Investigating the occurrence of different Anaplasma species in ticks from domestic and wild hosts improves the knowledge of tick-borne diseases in the Mediterranean area, and has implications in defining vector species distribution and risk assessment.

Molecular typing and diagnosis of Anaplasma spp. closely related to Anaplasma phagocytophilum in ruminants from Tunisia.

Accurate diagnosis of animal and zoonotic diseases, such as granulocytic anaplasmosis, is crucial to estimate risk during control programs. In this study, 16S rRNA nested PCR and RFLP assay were combined to investigate the presence of Anaplasma phagocytophilum and genetically related strains (namely A. phagocytophilum-like 1 and 2) in 936 Tunisian ruminants. By using this method, A. phagocytophilum was not detected in any of the tested animals, while A. phagocytophilum-like 1 and A. phagocytophilum-like 2 were detected at variable prevalence rates in sheep, goats and cattle at coinfection rates respectively of 3.9, 2.5 and 0.5%. Sequence analysis validated RFLP data, and confirmed the co-occurrence of two potentially novel species closely related to A. phagocytophilum in Tunisian ruminants. Phylogeny indicated the presence of genetic variants shared by different ruminant species for each type of A. phagocytophilum-like strains. Results raise concern on the use and interpretation of indirect and direct tests traditionally employed for detecting pathogenic A. phagocytophilum strains in ruminants and in other vertebrates' species, and provide additional background to improve classification of bacterial species closely related to A. phagocytophilum, and to reconstruct their evolutionary history.

Anaplasma platys-like strains in ruminants from Tunisia.

Molecular diagnosis of Anaplasma platys and related strains (A. platys-like) in carnivores and ruminants is challenging due to co-infections with cross-reacting strains, and require post-amplification sequencing of the hemi-nested PCR products traditionally generated by targeting the groEL gene. In this study, a Restriction Enzyme Fragment Length Polymorphism (RFLP) assay coupled to hemi-nested groEL PCR was developed to discriminate among A. platys and genetically related strains. This novel approach was used for investigating A. platys-like infection in 963 domesticated ruminants (241 goats, 355 sheep, and 367 cattle) from 22 delegations located in North Tunisia. Overall prevalence rates of A. platys-like were 22.8, 11, and 3.5% in goats, sheep, and cattle, respectively. Alignment, identity comparison, and phylogenetic analysis of the groEL sequence variants obtained in this study confirmed RFLP data suggesting that Tunisian ruminants are infected by novel unclassified Anaplasma strains genetically related to A. platys. Compared to sequencing, RFLP assay allows fast detection of A. platys and A. platys-like pathogens in the same sample and has a potential value especially when screening ticks, cats and ruminants, which can be a common host for these two bacteria. This newly developed molecular technique would provide valuable molecular tool for epidemiological studies related to A. platys as well as remove concern over specificity of serological and molecular methods routinely used to identify diverse Anaplasma strains and species in wild and domestic ruminants.

Characterization of Anaplasma spp. infection in dogs from Costa Rica.

A cross-sectional study combining serological and molecular techniques for detecting selected Anaplasma species was conducted between 2011 and 2012 in dogs and ticks sampled in all provinces of Costa Rica. Global Anaplasma spp. seroprevalence was 2.7% (11/408) by indirect immunofluorescence assay. The 16S rRNA PCR confirmed active A. phagocytophilum infection only in one dog (0.3%, 1/374); however, the same sample was negative to groEL PCR. Out of 122 Rhipicephalus sanguineus s.l. ticks analyzed, one (0.8%) was found positive to A. phagocytophilum 16S rRNA PCR but negative when tested by groEL PCR; this tick was collected from a seronegative and PCR negative dog. Both 16S rRNA sequences were 100% (510/510bp) identical to A. phagocytophilum strains isolated in different countries from different hosts. The presence of A. platys was established in four dogs (1%, 4/374) by both 16SrRNA and groEL PCR. Ticks collected from the same dogs tested negative by PCR. The 16S rRNA sequences were 100% identical to the corresponding sequences of A. platys strains isolated from dogs in Croatia and Brazil, however groEL sequences showed variable similarity levels (99-100%) with different strains of A. platys isolated in Chile, Japan and Thailand, pointing out the possible presence of different variants in Central America. Collectively data indicate low prevalence of A. phagocytophilum and A. platys in dogs from Costa Rica. Furthermore, infections seem to occur without clinical signs but with some hematological changes, and seem to resolve without treatment.

A case of canine neurological leishmaniasis.

In this study we describe a case of neurological leishmaniasis in a dog, reported in an endemic area, with signs of multifocal intracranial lesions involving the vestibular system and the cerebellum. Serum biochemistry revealed a decrease of albumin-globulin ratio caused by an increase of alfa2-, beta-, and gamma- globulin, while antibody titers were highly positive for Leishmania spp. Magnetic resonance images of the brain were consistent with a granulomatous in ammation/infection. Cerebrospinal fluid revealed a marked mononuclear pleocytosis and was positive to the Pandy Test, as well as to a Leishmania spp. -speci c polymerase chain reaction (PCR) assay. Sequencing of the PCR products revealed the highest similarity with several strains belonging to the Leishmania infantum / Leishmania chagasi group. Clinical response to treatment for systemic leishmaniasis was supportive of diagnosis. This report focuses on an atypical form of canine leishmaniasis and suggests that in endemic geographic areas leishmaniasis has to be considered for di erential diagnosis in neurological manifestations. Also, cerebrospinal liquor should always be tested when neurological symptoms are present in order to di erentiate neurological leishmaniasis from unrelated neurological signs in Leishmania positive patients.