Recently, the incidence and awareness of tick-borne diseases in humans and animals have increased due to several factors, which in association favor the chances of contact among wild animals and ...their ectoparasites, domestic animals and humans. Wild and domestic carnivores are considered the primary source of tick-borne zoonotic agents to humans. Among emergent tick-borne pathogens, agents belonging to family Anaplasmataceae (Order Rickettsiales) agents stand out due their worldwide distribution and zoonotic potential. In this review we aimed to review the genetic diversity of the tick-transmitted genera
Neoehrlichia sp." in wild carnivores Caniformia (Canidae, Mustelidae and Ursidae) and Feliformia (Felidae, Hyanidae, Procyonidae and Viverridae) worldwide, discussing the implications for human and domestic animal health and wildlife conservation. Red foxes (
) have been identified as hosts for
spp. (
),
and "
Neoehrlichia sp." (FU98 strain) and may contribute to the maintaenance of
in Europe. Raccoons (
) have been reported as hosts for
, "
Neoehrlichia lotoris" and
, and play a role in the maintenance of
in the USA. Raccoon dogs (
) may play a role as hosts for
and
. New
and/or
genotypes circulate in wild canids and felids from South America and Africa. While
sp. closely related to
has been reported in wild felids from Brazil and Japan,
sp. closely related to
has been detected in wild felids from Brazil and Africa. Red foxes and mustelids (otters) are exposed to
in countries located in the Mediaterranean basin, probably as a consequence of spillover from domestic dogs. Similarly,
occurs in procyonids in North (raccoons in USA, Spain) and South (
in Brazil) Hemispheres, in areas where
is frequent in dogs. While "
Neoehrlichia lotoris" seems to be a common and specific agent of raccoons in the USA, "
Neoehrlichia sp." (FU98 strain) seems to show a broader range of hosts, since it has been detected in red fox, golden jackal (
) and badger (
) in Europe so far. Brown (
) and black (
) bears seem to play a role as hosts for
in the North Hemisphere.
has been detected in wild Procyonidae, Canidae and Felidae in Asia and Brazil. In order to assess the real identity of the involved agents, future works should benefit from the application of MLST (Multi Locus Sequence Typing), WGS (Whole Genome Sequencing) and NGS (Next Generation Sequencing) technologies aiming at shedding some light on the role of wild carnivores in the epidemiology of Anaplasmataceae agents.
Rationale
Discovery proteomics has been popularized to be essential in the investigator's biological toolbox. Many biological problems involve the interplay of multiple organisms. Herein, a bottom‐up ...proteomics workflow was developed to study a system containing multiple organisms to promote a thorough understanding of how each interacts with the others.
Methods
A label‐free quantification proteomics workflow was developed with nanoscale liquid chromatography coupled to tandem mass spectrometry (nanoLC‐MS/MS). This protocol describes a bottom‐up proteomics workflow used to study differential protein expression in the context of fleas (Ctenocephalides felis felis) experimentally infected by the bacterium Bartonella henselae, the etiological agent of Cat Scratch Disease (CSD).
Results
Step‐by‐step instructions are provided for protein extraction, protein cleanup, total protein measurement, nanoLC‐MS/MS data acquisition, and data analysis using Proteome Discoverer software. Comprehensive and exhaustive details are included to promote the adoption of this proteomics workflow in other laboratories.
Conclusion
A proteomics protocol is detailed for a system containing multiple proteomes from different taxonomic lineages using CSD (cats bitten by fleas infected with Bartonella henselae) as a model. The operating protocol can be readily applied to other label‐free proteomics work involving multiple proteomes from taxonomically distinct organisms.
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The family Streblidae comprises a monophyletic group of Hippoboscoidea, hematophagous dipterans that parasitize bats. Bartonella spp. and Rickettsia spp. have been reported in bats sampled in Europe, ...Africa, Asia, North, Central and South America. However, there are few reports on the Bartonella and Rickettsia bacteria infecting Hippoboscoidea flies and mites. While Spinturnicidae mites are ectoparasites found only in bats, those belonging to the family Macronyssidae comprise mites that also parasitize other mammal species. This study investigates the occurrence and assesses the phylogenetic positioning of Bartonella spp. and Rickettsia spp. found in Streblidae flies and Spinturnicidae and Macronyssidae mites collected from bats captured in Brazil. From May 2011 to April 2012 and September 2013 to December 2014, 400 Streblidae flies, 100 Macronyssidaes, and 100 Spinturnicidae mites were collected from bats captured in two sites in northeastern Nova Iguaçu, Rio de Janeiro, southeastern Brazil. Forty (19.8%) out of 202 Streblidae flies were positive for Bartonella spp. in qPCR assays based on the nuoG gene. Among the flies positive for the bacterium, six (18%) were Paratrichobius longicrus, seven (29%) Strebla guajiro, two (40%) Aspidoptera phyllostomatis, five (11%) Aspidoptera falcata, one (10%) Trichobius anducei, one (25%) Megistopoda aranea, and 18 (32%) Trichobius joblingi, and collected from bats of the following species: Artibeus lituratus, Carollia perspicillata, Artibeus planirostris, Sturnira lilium, and Artibeus obscurus. Six sequences were obtained for Bartonella (nuoG n = 2, gltA n = 2, rpoB n = 1, ribC = 1). The phylogenetic analysis based on gltA (750pb) gene showed that the Bartonella sequences clustered with Bartonella genotypes detected in bats and ectoparasites previously sampled in Latin America, including Brazil. Only one sample (0.49%) of the species Trichobius joblingi collected from a specimen of Carollia perspicillata was positive for Rickettsia sp. in cPCR based on the gltA gene (401bp). This sequence was clustered with a 'Candidatus Rickettsia andaenae" genotype detected in an Amblyomma parvum tick collected from a rodent in the southern region of Brazilian Pantanal. The sampled Macronyssidae and Spinturnicidae mites were negative for Bartonella spp. and Rickettsia spp. This study demonstrated the first occurrence of Bartonella spp. and Rickettsia spp. DNA in Streblidae flies collected from bats in Brazil.
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Feline cytauxzoonosis is a disease caused by Cytauxzoon felis, a protozoan that infects the red blood cells and macrophages. It is responsible for an acute and often fatal disease in domestic cats. ...The purpose of this study was to investigate the occurrence of C. felis infections in healthy cats. Piroplasm forms were seen in the erythrocytes of 2 cats, and C. felis DNA was identified by polymerase chain reaction (PCR) in one of them. The results demonstrate that erythrocytic piroplasmids associated with tick-borne parasitic protozoa may be found circulating in the blood of healthy cats in Rio de Janeiro. These can be differentiated from the morphologically similar forms of species such as Babesia by analysis of DNA, thereby demonstrating the potential for further studies of feline populations in Brazil.
Piroplasmida is an order of the phylum Apicomplexa that comprises the
Babesia
,
Cytauxzoon
, and
Theileria
genera. These hemoparasites infect vertebrate blood cells and may cause serious diseases in ...animals and humans. Even though previous studies have shown that bats are infected by different species of piroplasmids, the occurrence and diversity of these hemoparasites have not been investigated in this group of mammals in Brazil. Therefore, the present work aimed to investigate the occurrence and assess the phylogenetic placement of piroplasmids infecting bats sampled in a peri-urban area from Central-Western Brazil. Seventeen (12.6%) out of 135 animals were positive by nested PCR assay for the detection of
Babesia/Theileria
targeting the 18S rRNA gene. Eleven sequences of the 17 positive samples could be analyzed and showed an identity of 91.8–100% with
Theileria bicornis
,
Babesia vogeli
, a
Babesia
sp. identified in a small rodent (
Thrichomys pachyurus
) from the Brazilian Pantanal and a
Babesia
sp. identified in a dog from Thailand as assessed by nBLAST. A phylogenetic tree was constructed from an alignment of 1399 bp length using analyzed and known piroplasmid 18S rRNA sequences. In this tree, piroplasmid 18S rRNA sequences detected in three specimens of
Phyllostomus discolor
(Piroplasmid n. sp.,
P. discolor
) were placed as a sister taxon to
Theileria
sensu stricto (Clade V) and
Babesia
sensu stricto (Clade VI). An additional phylogenetic tree was generated from a shorter alignment of 524 bp length including analyzed piroplasmid 18S rRNA sequences of bat species
Artibeus planirostris
and
A. lituratus
(Piroplasmid sp.,
Artibeus
spp.). The two 18S rRNA sequences detected in
Artibeus
spp. (Piroplasmid n. sp.,
Artibeus
spp.) were placed within
Babesia
sensu stricto (Clade VI) into a strongly supported clade (bootstrap: 100) that included
Babesia vogeli
. The two 18S rRNA sequences of Piroplasmid sp.,
Artibeus
spp. showed a single and a two-nucleotide differences, respectively, with respect to
B. vogeli
in a 709 pb length alignment. For the first time, the present study shows the occurrence of putative new piroplasmid species in non-hematophagous bats from Brazil.
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The occurrence of Trypanosoma spp. in wild carnivore populations has been intensively investigated during the last decades. However, the impact of these parasites on the health of free-living ...infected animals has been largely neglected. The Pantanal biome is the world's largest seasonal wetland, harboring a great diversity of species and habitats. This includes 174 species of mammals, of which 20 belong to the order Carnivora. The present study aimed to investigate the effect of Trypanosoma evansi and Trypanosoma cruzi infections and coinfections on the health of the most abundant carnivores in the Pantanal: coati (Nasua nasua), crab-eating fox (Cerdocyon thous), and ocelot (Leopardus pardalis). We captured 39 coatis, 48 crab-eating foxes, and 19 ocelots. Diagnostic tests showed T. cruzi infection in 7 crab-eating foxes and 5 coatis. Additionally, 7 crab-eating foxes, 10 coatis, and 12 ocelots were positive for T. evansi. We observed coinfections in 9 crab-eating foxes, 8 coatis, and 2 ocelots. This is the first report of T. evansi and T. cruzi infection on the health of free-living ocelots and crab-eating foxes. We showed that single T. evansi or T. cruzi infection, as well as coinfection, caused some degree of anemia in all animals, as well as an indirect negative effect on body condition in coatis and crab-eating foxes via anemia indicators and immune investment, respectively. Furthermore, the vigorous immune investment observed in sampled coatis, crab-eating foxes and ocelots infected by T. evansi, T. cruzi and coinfected can be highly harmful to their health. Overall, our results indicate that single and combined infection with T. evansi and T. cruzi represent a severe risk to the health of wild carnivores in the Pantanal region.
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This study reports the occurrence of parasites belonging to the
Hepatozoon
genus in fish
Hoplias aimara
from the Eastern Amazon. Fish (
n
= 54) were sampled from the Falsino River, located in the ...Amapá National Forest (FLONA), in the state of Amapá, northern Brazil. Fresh liver preparations were examined in the field between a slide and a coverslip under a light microscope. Cysts containing
Hepatozoon
cystozoites were observed in the liver of 5 (9%) out of 54
H. aimara
individuals. The cysts were ovoid (mean dimensions 10.28 × 9.8 μm), presenting up to four elongated cystozoites (mean dimensions 11.04 × 1.68 μm), containing 1 to 4 residual bodies of different sizes. A single liver sample containing cysts was submitted to DNA extraction and PCR analyses based on a fragment of the 18S rRNA gene. The sequencing revealed a 465 bp fragment exhibiting 99% query coverage, 0.0
E
-value, and 98.7% identity with
Hepatozoon caimani
(MF322538 and MF322539), detected in caimans (
Caiman yacare
) from Brazil. This is the first report of the occurrence of cysts containing
Hepatozoon
cystozoites in free-living fishes.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
Feline piroplasmids include the genera
Babesia
spp.,
Cytauxzoon
spp., and
Theileria
spp. In Brazil, there are few reports regarding these hemoprotozoans; however, clinicopathological and molecular ...data are scarce. This study aimed to characterize the clinical relevance of these parasites through hematological, biochemical, and molecular approaches. For this purpose, 166 cats from Brasilia, Federal District, Midwestern Brazil, were screened using a quantitative polymerase chain reaction (qPCR) for piroplasmids based on the LSU4 mitochondrial gene, which resulted in an overall prevalence of 36/166 (21.7%). Twelve of 166 samples (7.2%) were positive for
C. felis
, while 19/166 (11.4%) were positive for
Babesia vogeli
. No samples tested positive for
Theileria
spp.
Babesia vogeli
and
Cytauxzoon
spp
.
LSU4 sequences showed identities of 97–100% and 99.3%, respectively, to US isolates. The hematological and biochemical findings did not differ significantly between the cats that tested positive and negative for piroplasmids. Although the lack of abnormalities in clinical and laboratory parameters does not eliminate the possibility that these cats were sick and recovered, it may suggest that the Brazilian strain of
Cytauxzoon
spp. is not as pathogenic as that from the USA, despite the high molecular identity with North American isolates.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
The present work aimed to investigate the genetic diversity of Bartonella in mammals and ectoparasites in Pantanal wetland, Brazil. For this purpose, 31 Nasua nasua, 78Cerdocyon thous, 7Leopardus ...pardalis, 110 wild rodents, 30 marsupials, and 42 dogs were sampled. DNA samples were submitted to a quantitative real-time PCR assay (qPCR). Positive samples in qPCR were submitted to conventional PCR assays targeting other five protein-coding genes. Thirty-five wild rodents and three Polygenis (P.) bohlsi bohlsi flea pools showed positive results in qPCR for Bartonella spp. Thirty-seven out of 38 positive samples in qPCR were also positive in cPCR assays based on ftsZ gene, nine in nuoG-cPCR, and six in gltA-cPCR. Concatenated phylogenetic analyses showed that two main genotypes circulate in rodents and ectoparasites in the studied region. While one of them was closely related to Bartonella spp. previously detected in Cricetidae rodents from North America and Brazil, the other one was related to Bartonella alsatica, Bartonella pachyuromydis, Bartonella birtlesii, Bartonella acomydis, Bartonella silvatica, and Bartonella callosciuri. These results showed that at least two Bartonella genotypes circulate among wild rodents. Additionally, the present study suggests that Polygenis (P.) bohlsi bohlsi fleas could act as possible Bartonella vectors among rodents in Pantanal wetland, Brazil.
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