Leprosy is an infectious disease caused by Mycobacterium leprae and frequently resulting in irreversible deformities and disabilities. Ticks play an important role in infectious disease transmission ...due to their low host specificity, worldwide distribution, and the biological ability to support transovarial transmission of a wide spectrum of pathogens, including viruses, bacteria and protozoa. To investigate a possible role for ticks as vectors of leprosy, we assessed transovarial transmission of M. leprae in artificially-fed adult female Amblyomma sculptum ticks, and infection and growth of M. leprae in tick cell lines. Our results revealed M. leprae RNA and antigens persisting in the midgut and present in the ovaries of adult female A. sculptum at least 2 days after oral infection, and present in their progeny (eggs and larvae), which demonstrates the occurrence of transovarial transmission of this pathogen. Infected tick larvae were able to inoculate viable bacilli during blood-feeding on a rabbit. Moreover, following inoculation with M. leprae, the Ixodes scapularis embryo-derived tick cell line IDE8 supported a detectable increase in the number of bacilli for at least 20 days, presenting a doubling time of approximately 12 days. As far as we know, this is the first in vitro cellular system able to promote growth of M. leprae. Finally, we successfully transformed a clinical M. leprae isolate by inserting the reporter plasmid pCHERRY3; transformed bacteria infected and grew in IDE8 cells over a 2-month period. Taken together, our data not only support the hypothesis that ticks may have the potential to act as a reservoir and/or vector of leprosy, but also suggest the feasibility of technological development of tick cell lines as a tool for large-scale production of M. leprae bacteria, as well as describing for the first time a method for their transformation.
IDE8 tick cell cultures have been used for the isolation and propagation of several isolates of Anaplasma marginale. The genetic heterogeneity of A. marginale strains in cattle is diverse in endemic ...regions worldwide and the analyses of msp1α (major surface protein 1 alpha) gene sequences have allowed the identification of different strains. This study reports the isolation and propagation of two new isolates of A. marginale in IDE8 cells from blood of two cattle and their morphological and molecular characterization using light microscopy and the msp1α gene, respectively. Small colonies were observed in cytospin smears of each of the isolates 60 days after culture initiation. Based on msp1α sequence variation, the two isolates were found to be separate strains and were named AmRio1 and AmRio2. Analysis of msp1α microsatellite in both strains resulted in a single genotype, genotype E. The amino acid sequence of one MSP1α tandem repeat from the strain AmRio1 resulted in a new sequence (named 162) with one amino acid change. The results of these phylogenetic analyses demonstrated that A. marginale strains from Brazil and Argentina formed two large clusters of which one was less divergent that the other.
Resumo: A espiroquetose aviária é uma enfermidade septicêmica de curso agudo, cosmopolita, que acomete diversas espécies aviárias, causada por Borrelia anserina e transmitida pelo carrapato Argas ...miniatus. O experimento teve como objetivos avaliar as alterações bioquímicas e anátomo-histopatológicas no fígado de Gallus gallus, causadas pela infecção experimental por B. anserina. Quarenta aves da espécie G. gallus foram divididas em quatro grupos inteiramente casualizados com 10 animais cada: G1 - inoculado com soro infectado com B. anserina; G2 - inoculado com soro fisiológico a 0,9%; G3 - exposto a ninfas de terceiro ínstar de A. miniatus infectados por B. anserina; G4 - exposto a ninfas de terceiro ínstar de A. miniatus livres de B. anserina. As aves dos Grupos 1 e 3 manifestaram no 3º e 6º dias pós-inoculação (DPI) respectivamente, sintomatologia característica da doença como inapetência, perda de peso, sonolência, diarreia esverdeada, mucosas hipocoradas, penas arrepiadas e hipertermia. Os níveis de ALT do Grupo 1 mostraram-se significativamente mais elevados apenas no 12ºDPI e 24ºDPI em relação ao seu grupo controle (Grupo 2) e no Grupo 3 esses níveis se mantiveram elevados até o 20º DPI em comparação ao seu grupo controle (Grupo 4). Os níveis da enzima AST pouco oscilaram nos grupos experimentais, embora tenham sido encontradas elevações no 12ºDPI nos Grupos 1 e 3. Os fígados das aves dos Grupos 1 e 3 apresentaram à necropsia, moderada hepatomegalia, congestão, superfície irregular e coloração vermelha a cianótica; constataram-se ainda pequenos pontos esbranquiçados na superfície. A histopatologia do fígado revelou congestão, infiltrados inflamatórios mononucleares, focos de necrose fibrinoide, dilatação dos sinusoides e vacuolização de hepatócitos. A coloração de Warthin-Starry revelou, nos fígados das aves dos Grupos 1 e 3, a presença de espiroquetas compatíveis com B. anserina, frequentemente no interior de vasos sanguíneos.
Resumo: A espiroquetose aviária é uma enfermidade septicêmica de curso agudo, cosmopolita, que acomete diversas espécies aviárias, causada por Borrelia anserina e transmitida pelo carrapato Argas ...miniatus. O experimento teve como objetivos avaliar as alterações bioquímicas e anátomo-histopatológicas no fígado de Gallus gallus, causadas pela infecção experimental por B. anserina. Quarenta aves da espécie G. gallus foram divididas em quatro grupos inteiramente casualizados com 10 animais cada: G1 - inoculado com soro infectado com B. anserina; G2 - inoculado com soro fisiológico a 0,9%; G3 - exposto a ninfas de terceiro ínstar de A. miniatus infectados por B. anserina; G4 - exposto a ninfas de terceiro ínstar de A. miniatus livres de B. anserina. As aves dos Grupos 1 e 3 manifestaram no 3º e 6º dias pós-inoculação (DPI) respectivamente, sintomatologia característica da doença como inapetência, perda de peso, sonolência, diarreia esverdeada, mucosas hipocoradas, penas arrepiadas e hipertermia. Os níveis de ALT do Grupo 1 mostraram-se significativamente mais elevados apenas no 12ºDPI e 24ºDPI em relação ao seu grupo controle (Grupo 2) e no Grupo 3 esses níveis se mantiveram elevados até o 20º DPI em comparação ao seu grupo controle (Grupo 4). Os níveis da enzima AST pouco oscilaram nos grupos experimentais, embora tenham sido encontradas elevações no 12ºDPI nos Grupos 1 e 3. Os fígados das aves dos Grupos 1 e 3 apresentaram à necropsia, moderada hepatomegalia, congestão, superfície irregular e coloração vermelha a cianótica; constataram-se ainda pequenos pontos esbranquiçados na superfície. A histopatologia do fígado revelou congestão, infiltrados inflamatórios mononucleares, focos de necrose fibrinoide, dilatação dos sinusoides e vacuolização de hepatócitos. A coloração de Warthin-Starry revelou, nos fígados das aves dos Grupos 1 e 3, a presença de espiroquetas compatíveis com B. anserina, frequentemente no interior de vasos sanguíneos.
Abstract: Spirochetosis avian is a septicemic disease of acute course and cosmopolitan can affect various avian species, caused by Borrelia anserina and transmitted by Argas miniatus. The experiment aimed to evaluate the biochemical, anatomical and histopathological changes in the liver of Gallus gallus caused by experimental infection with B. anserina. A total of 40 fowls of the species G. gallus were divided into four randomized groups of ten fowls each: G1 - inoculated with serum infected with B. anserina; G2 - inoculated with 0.9% saline; G3 - exposed to nymphs of 3rd instar of A. miniatus infected with B. anserina; G4 - exposed to ticks nymphs of 3rd instar of A. miniatus free of B. anserina. The fowls of Groups 1 and 3 expressed at 3 and 6 days post-inoculation (DAI) respectively , symptoms characteristic of the disease as lack of appetite , weight loss , drowsiness, greenish diarrhea, pale mucous membranes , ruffled feathers and hyperthermia. ALT of group 1 levels were significantly higher only at the 12º and 24º day after inoculation (DAI) compared with its control group (group 2), and in group 3 these levels remained high until the 20º DAI as compared with its control group (group 4). AST enzyme fluctuated little in the experimental groups, although elevations at 12ºDAI has been found in group 1 and 3. The liver of fowls in groups 1 and 3, presented at necropsy moderate hepatomegaly, congestion, irregular surface and red color to cyanotic. If found even small whitish spots on the surface. The histopathology revealed congestion, mononuclear inflammatory infiltrates, fibrinoid necrotic foci, dilatation of sinusoids, and vacuolation of hepatocytes. The Warthin-Starry staining revealed in the liver of fowls in groups 1 and 3 the presence of spirochetes compatible with B. anserina, often within blood vessels.
Sera samples were collected from 330 water buffaloes on Marajó Island, state of Pará, Brazil, to assess the presence of antibodies against Borrelia burgdorferi by indirect Enzyme Linked Immunosorbent ...Assay. Approximately 45% of the animals had antibodies against B. burgdoferi. The prevalence of seropositive buffaloes, 72% (85/118), was statistically higher in the city of Soure than in the other municipalities tested. Murrah breed animals were significantly more seropositive (Prevalence Ratio (PR) = 1.84, p = 0.000) than those of the Mediterranean breed. Among the animals diagnosed positive for tuberculosis, 33% (4/12), were also seropositive for B. burgdoferi. Animals positive for tuberculosis had a significantly lower level of B. burgdorferi seropositivity (PR = 1.36, p = 0.0017) than negative animals. The study showed that there was an agent immunologically close to B. burgdorferi circulating in water buffalo on Marajó Island, state of Pará, Brazil. These results demonstrate a possible role of the water buffalo in the epidemiological chain of this agent in Brazil, and are a warning of the danger this disease could pose to public health.
The in vitro feeding of ticks facilitates the conduction of studies involving the intrinsic vector-pathogen relationship, susceptibility tests, and resistance to acaricides, in addition to mimicking ...the use of experimental hosts. The objective of this study was to establish an in vitro feeding system using silicone membranes to supply various diets to the species
Ornithodoros rostratu
s. Each experimental group included 130 first-instar
O. rostratus
nymphs. The groups were divided according to the diet provided: citrated rabbit blood, citrated bovine blood, bovine blood with antibiotics, and defibrinated bovine blood. The control group was fed directly on rabbits. Ticks were weighed before and after the feeding and monitored individually according to their biological parameters. The results of the experiment demonstrated that the proposed system was efficient in terms of fixation stimulus and satisfactory in terms of tick engorgement, which would allow the maintenance of
O. rostratus
colonies by using artificial feeding through silicone membranes. All diets provided were efficient for the maintenance of colonies, but the ticks that received citrated rabbit blood displayed similar biological parameters to those observed under in vivo feeding conditions.
The present study aimed to use polymerase chain reaction (PCR) to detect species of the order Piroplasmida, such as Anaplasma spp., Borrelia spp., and Ehrlichia spp., circulating in the blood of ...Didelphis aurita in a peridomiciliary environment. Blood samples collected from big-eared opossum (Didelphis aurita) were screened for hemoparasites using PCR. The extracted DNA was tested for tick-borne hemoparasites. We were unable to detect hemoparasites, such as Ehrlichia spp., Babesia spp., Anaplasma spp., and Borrelia spp. Theileria DNA was detected in only one sample screened using PCR for an approximately 650-base pair fragment of the 18S rRNA gene. Sequencing and BLAST analysis of a subset of the PCR amplicons revealed 97% (535/553 bp) identity with Theileria bicornis. The detection of Theileria sp. in D. aurita challenges us to pursue more in-depth studies of marsupial piroplasmosids and to evaluate the morphological aspects of the findings and their possible involvement in zoonoses.
Abstract The present study aimed for the molecular detection of Rickettsia spp. in ticks collected from military training areas in southeastern Brazil. Previously, a total of 9,374 free-living ticks ...in 66 military instruction areas of five Military Organizations (MOs) were collected. From these, a sample of 1,885 ticks was used for molecular analysis..DNA extraction was performed using the “HotSHOT” technique, and PCR was performed using the Rickettsia spp. gltA, 17kDa and ompA genes. It was found that 11 samples produced DNA sequences of sizes compatible with the Rickettsia spp. gltA and 17kDa gene and two of these, also for ompA, with a prevalence of 0.58% (11/1885). Analyses of amplified fragments of gltA identified the samples as Rickettsia bellii in seven nymphs of Amblyomma dubitatum and one nymph of Amblyomma sculptum, and identified samples as Rickettsia sp. in two nymphs of A. dubitatum and one of A. sculptum. This finding warns the possibility of the risk of exposure of military personnel to ticks infected with Rickettsia spp. during training.
Experimental infection by Borrelia anserina strain PL in Gallus gallus Cepeda, Marcio Barizon; Cordeiro, Matheus Dias; Baêta, Bruna de Azevedo ...
Semina. Ciências agrárias : revista cultural e científica da Universidade Estadual de Londrina,
07/2021, Letnik:
42, Številka:
4
Journal Article
Recenzirano
Odprti dostop
Avian borreliosis is a tick-borne acute septicemic disease that affects a variety of avian species and is caused by the bacterium Borrelia anserina. The present study aimed to establish the infection ...dynamics of B. anserina in experimentally infected Gallus gallus to determine its impact on the health and weight gain of the animals. Forty G. gallus were divided into four groups. Group 1 (G1) included 10 animals inoculated intramuscularly (IM) with 250μL of blood serum containing 3.7x106 spirochetes per milliliter. Group 2 (G2) included 10 animals inoculated IM with 250μL of saline 0.9%. Group 3 (G3) included 10 animals infested with four Argas miniatus nymphs (3rd instar) infected with B. anserina strain PL. Group 4 (G4) included 10 animals infested with four A. miniatus nymphs (3rd instar) free of infection by B. anserina. The number of spirochetes in the blood was directly proportional to the appearance of clinical signs and symptoms. This is the first study showing the experimental clinical evolution of the B. anserina PL strain in G. gallus. Additionally, the first assessments of spirochetaemia are presented, based on the direct count of bacteria in the serum by a Neubauer chamber, during the infection.
Human contact with wild animals in synanthropic habits is often mediated by arthropod vectors such as ticks. This is an important method of spreading infectious agents that pose a risk to human ...health. Thus, this study aimed to molecularly detect
Ehrlichia
spp.,
Anaplasma
spp.,
Borrelia
spp., and protozoa of the order Piroplasmida in ticks collected from coatis of Iguaçu National Park (PNI), Paraná, Brazil. This study involved 553 ticks DNA, including
Amblyomma
spp. larvae,
Haemaphysalis juxtakochi
nymphs,
Amblyomma brasiliense
,
Amblyomma coelebs
, and adults of
Amblyomma ovale
. The DNA extracted from each sample was subjected to polymerase chain reaction (PCR) targeting the genes 23S rRNA for the Anaplasmataceae family, 16S rRNA for
Anaplasma
spp.,
dsb
for
Ehrlichia
spp.,
flaB
, 16S rRNA,
hpt
, and
glpQ
for
Borrelia
spp., and 18S rRNA for Piroplasmid protozoans. DNA from
Anaplasma
sp. was detected in ticks of the species
A. coelebs
(4/553);
Borrelia
sp. DNA was detected in
A. coelebs
(3/553),
A. ovale
(1/553), and
Amblyomma
larvae (1/553); and
Theileria
sp. was detected in
A. coelebs
(2/553). All tested samples were negative for
Ehrlichia
spp. Our study constitutes the newest report in South America of these microorganisms, which remain poorly studied.
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