RESUMO
INTRODUCTION: Hookworm infections are neglected tropical diseases of humans and animals worldwide. A meta-analysis and systematic review was designed to evaluate the prevalence of hookworm infection in animal and human hosts in Asia until July 2018. METHODS: The available online articles of five English databases (PubMed, Scopus, Science Direct, Web of Science and Google Scholar) were explored. RESULTS: The most parasitized carnivores were jackal (48%, CI: 4 to 91%), followed by dog (41%, CI: 29 to 53%), cat (26%, CI: 14 to 38%) and the red fox (19%, CI: 13 to 24%). The weighted prevalence of Ancylostoma braziliensis, A. caninum, A. ceylanicum, A. tubaeforme and Uncinaria stenocephala isolated from different canids were found to be 27% (CI: 21 to 33%), 23% (CI: 7.0 to 53%), 24% (CI: 12 to 35%), 44% (CI: 37 to 51%) and 37% (CI: 18 to 55%), respectively. In total, 98 records were obtained for human hookworms from 3209 760 examined individuals and the calculated weighted prevalence in this population was 19% (CI: 17 to 20%). CONCLUSIONS: These findings highlight a desirable ecological milieu for parasite survival and transmission in such territories, which implicates revisiting control programs and public health infrastructures in those areas.
Assuntos
Infecções por Uncinaria/veterinária , Ancylostoma/classificação , Animais , Ásia/epidemiologia , Doenças do Gato/epidemiologia , Doenças do Gato/parasitologia , Gatos , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Cães , Fezes , Raposas , Infecções por Uncinaria/epidemiologia , PrevalênciaRESUMO
BACKGROUND: The immunomodulatory role of many parasites is well-documented. The current study designed to assess the immunoregulatory effects of the somatic extract (SE) of Toxocara canis on murine model of airway inflammations. METHODS: The experiment was performed in department of parasitology of Tarbiat Mo-dares University, Tehran, Iran from November 2018 to May 2019. Totally 30 female BALB/c mice divided into one control group and two experimental groups (10 mice in each group). The ovalbumin (OVA) group was sensitized with OVA in alum, while the SE group was administered with SE and OVA in alum intraperitoneally. The control group was injected with PBS in alum. Then, SE and OVA groups were intranasally challenged with OVA for three consecutive days and the control group encountered with PBS at the same time. One day after the last challenge, real-time PCR and histopathology survey were conducted on isolated lung tissues. RESULTS: The gene expression of IL-25, IL-33, TNF-α and TLR-4 in SE group was significantly lower than OVA group (P<0.05). The level of IL-10, TGF-ß and IFN-γ were considerably higher than the OVA group (P<0.05). The inflammation was reduced in SE group, as the total cell number of bronchoalveolar lavage fluid was less than OVA group. Based on the histopathology findings the inflammation was decreased in SE group compared to the OVA group. CONCLUSION: Although, an inhibitory effect of SE of T. canis on airway inflammations was detected, there is still a long way ahead regarding the indication of the precise mechanisms.
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Despite the high prevalence of canine Leishmania infantum infection in Portugal, significant differences associated with different risk factors can be found between geographically contiguous areas. In this study, a geographical area within the central region of Portugal (municipalities of Proença-a-Nova, Mação and Vila de Rei) was investigated. An epidemiological survey involved the analysis by an indirect enzyme-linked immunosorbent assay of serum samples collected during the anti-rabies vaccination campaign from 282 dogs. Geospatial analysis showed the distribution of geospatial prevalence of leishmaniosis and has delimited two areas (clusters) with a statistically significant higher risk of seropositivity in dogs (pâ¯=⯠0.003 and p = 0.027, for clusters 1 and 2, respectively). The highest seroprevalence (56.0%; CI: 41.2-70.0) was found in Vila de Rei. Five land occupation types showed a possible influence on the geographic distribution of seropositivity, with statistically significant differences between seropositive and seronegative dogs. Land occupied by temporary irrigated crops (pâ¯=⯠0.026), olive groves (pâ¯=⯠0.013), complex cultural systems and parcelling (pâ¯=⯠0.021), open forests, logging and new plantations (pâ¯=⯠0.043) and watercourses (pâ¯=⯠0.012) influenced the geographical distribution of canine Leishmania infection. Seropositive dogs had a greater average area of occupied land (i.e. open forests, logging and new plantations) than the seronegative ones (3.1439â¯km2 versus 2.5650â¯km2, respectively; pâ¯=⯠0.043).
Assuntos
Doenças do Cão/parasitologia , Leishmaniose/veterinária , Animais , Doenças do Cão/epidemiologia , Cães , Humanos , Leishmaniose/epidemiologia , Leishmaniose/parasitologia , Portugal/epidemiologia , Fatores de Risco , Estudos SoroepidemiológicosRESUMO
BACKGROUND: Of increasing importance to the medical and veterinary communities is the zoonotic filarioid nematode Onchocerca lupi. Onchocercosis, thus far found in wolves, dogs, cats and humans, is diagnosed via skin snips to detect microfilariae and surgical removal of adults from the eye of the host. These methods are time-consuming, laborious and invasive, highlighting the need for new tools for the diagnosis of O. lupi in susceptible hosts. Symptoms related to the presence of the adults in the eye can range from none apparent to severe, including blindness. No reliable chemotherapeutic protocols are available, as yet, to eliminate the infection. Paramyosin, an invertebrate-specific protein, has been well-studied as an allergen, diagnostic marker and vaccine candidate. The aim of this study, therefore, was to isolate and characterise paramyosin from O. lupi to assess its suitability for the development of a serological diagnostic assay. METHODS: The adult and microfilarial stages of O. lupi were isolated from the eyes and skin of a 3-year-old male dog. Total RNA was extracted and reverse transcribed into single stranded cDNA. Reverse-transcription PCR was used to isolate a full-length paramyosin cDNA from adult worms and to investigate the temporal expression patterns of this gene. All amplicons were sequenced using dideoxy chain termination sequencing. Bioinformatics was used to predict the amino acid sequence of the gene, to compare the DNA and protein sequences with those available in public databases and to investigate the phylogenetic relationship of all molecules. Antibody binding sites were predicted using bioinformatics and mapped along with published antigenic epitopes against the O. lupi paramyosin protein. The native protein, and three smaller recombinantly expressed peptides, were subjected to western blot using serum from dogs both positive and negative for O. lupi. RESULTS: Paramyosin of O. lupi was herein molecularly characterized, encoded by a transcript of 2,643 bp and producing a protein of 881 amino acids (101.24 kDa). The paramyosin transcript was detected, by reverse transcription PCR, in adults and microfilariae, but not in eggs. Phylogenetic analysis indicates that this molecule clusters with paramyosins from other filarioids to the exclusion of those from other taxa. A total of 621 unique antibody binding epitopes were predicted for this protein and another 28 were conserved in other organisms. This information was used to design three peptides, for recombinant expression, to identify the antibody binding epitope(s) and reduce potential cross-reactivity with serum from dogs infected with other filarioid nematodes. Native paramyosin, purified from microfilariae and adults, was detected by antibodies present in serum from dogs with known O. lupi infections. CONCLUSIONS: Data provided herein may assist in the development of a serological diagnostic test, based on antibodies to O. lupi paramyosin, for the diagnosis of this infection, in order to gain more information on the real distribution of this little known filarioid of zoonotic concern.
Assuntos
Doenças do Cão/diagnóstico , Doenças Negligenciadas/diagnóstico , Onchocerca/química , Oncocercose Ocular/diagnóstico , Oncocercose/diagnóstico , Tropomiosina/genética , Tropomiosina/imunologia , Adulto , Animais , Gatos , Biologia Computacional , Doenças do Cão/parasitologia , Cães , Epitopos/imunologia , Humanos , Masculino , Microfilárias/genética , Microfilárias/isolamento & purificação , Doenças Negligenciadas/parasitologia , Onchocerca/imunologia , Onchocerca/isolamento & purificação , Oncocercose/sangue , Oncocercose/imunologia , Oncocercose/parasitologia , Oncocercose Ocular/sangue , Oncocercose Ocular/imunologia , Oncocercose Ocular/parasitologia , Filogenia , Reação em Cadeia da Polimerase , Testes Sorológicos , Tropomiosina/sangue , Tropomiosina/isolamento & purificação , ZoonosesRESUMO
BACKGROUND: Bovine besnoitiosis, caused by the protozoan Besnoitia besnoiti, reduces productivity and fertility of affected herds. Besnoitiosis continues to expand in Europe and no effective control tools are currently available. Experimental models are urgently needed. Herein, we describe for the first time the kinetics of standardised in vitro models for the B. besnoiti lytic cycle. This will aid to study the pathogenesis of the disease, in the screening for vaccine targets and drugs potentially useful for the treatment of besnoitiosis. METHODS: We compared invasion and proliferation of one B. tarandi (from Finland) and seven B. besnoiti isolates (Bb-Spain1, Bb-Spain2, Bb-Israel, Bb-Evora03, Bb-Ger1, Bb-France, Bb-Italy2) in MARC-145 cell culture. Host cell invasion was studied at 4, 6, 8 and 24 h post infection (hpi), and proliferation characteristics were compared at 24, 48, 72, 96, 120, and 144 hpi. RESULTS: In Besnoitia spp., the key parameters that determine the sequential adhesion-invasion, proliferation and egress steps are clearly distinct from those in the related apicomplexans Toxoplasma gondii and Neospora caninum. Besnoitia spp. host cell invasion is a rather slow process, since only 50 % of parasites were found intracellular after 3-6 h of exposure to host cells, and invasion still took place after 24 h. Invasion efficacy was significantly higher for Bb-France, Bb-Evora03 and Bb-Israel. In addition, the time span for endodyogeny to take place was as long as 18-35 h. Bb-Israel and B. tarandi isolates were most prolific, as determined by the tachyzoite yield at 72 hpi. The total tachyzoite yield could not be predicted neither by invasion-related parameters (velocity and half time invasion) nor by proliferation parameters (lag phase and doubling time (dT)). The lytic cycle of Besnoitia was asynchronous as evidenced by the presence of three different plaque-forming tachyzoite categories (lysis plaques, large and small parasitophorous vacuoles). CONCLUSIONS: This study provides first insights into the lytic cycle of B. besnoiti isolates and a standardised in vitro model that allows screening of drug candidates for the treatment of besnoitiosis.
Assuntos
Proliferação de Células , Células Epiteliais/parasitologia , Modelos Biológicos , Sarcocystidae/crescimento & desenvolvimento , Animais , Linhagem Celular , HaplorrinosRESUMO
The identification of populations and spatial genetic patterns is important for ecological and conservation research, and spatially explicit individual-based methods have been recognised as powerful tools in this context. Mammalian carnivores are intrinsically vulnerable to habitat fragmentation but not much is known about the genetic consequences of fragmentation in common species. Stone martens (Martes foina) and red foxes (Vulpes vulpes) share a widespread Palearctic distribution and are considered habitat generalists, but in the Iberian Peninsula stone martens tend to occur in higher quality habitats. We compared their genetic structure in Portugal to see if they are consistent with their differences in ecological plasticity, and also to illustrate an approach to explicitly delineate the spatial boundaries of consistently identified genetic units. We analysed microsatellite data using spatial Bayesian clustering methods (implemented in the software BAPS, GENELAND and TESS), a progressive partitioning approach and a multivariate technique (Spatial Principal Components Analysis-sPCA). Three consensus Bayesian clusters were identified for the stone marten. No consensus was achieved for the red fox, but one cluster was the most probable clustering solution. Progressive partitioning and sPCA suggested additional clusters in the stone marten but they were not consistent among methods and were geographically incoherent. The contrasting results between the two species are consistent with the literature reporting stricter ecological requirements of the stone marten in the Iberian Peninsula. The observed genetic structure in the stone marten may have been influenced by landscape features, particularly rivers, and fragmentation. We suggest that an approach based on a consensus clustering solution of multiple different algorithms may provide an objective and effective means to delineate potential boundaries of inferred subpopulations. sPCA and progressive partitioning offer further verification of possible population structure and may be useful for revealing cryptic spatial genetic patterns worth further investigation.
Assuntos
Carnívoros/genética , Ecossistema , Animais , Variação Genética , Humanos , Repetições de Microssatélites/genética , Especificidade da EspécieRESUMO
Trichinellosis is one of the most important foodborne parasitic zoonoses, caused by nematodes of the genus Trichinella. Pigs and other domestic and wild animals, including red foxes (Vulpes vulpes), are sources of Trichinella infection for human beings. Trichinella britovi is the major agent of infection in sylvatic animals and the most important species circulating in the European wildlife. The present study aimed at assessing Trichinella spp. infection in red foxes from the North of Portugal. Forty-seven carcasses of wild red foxes shot during the official hunting season or killed in road accidents were obtained between November 2008 and March 2010. In order to identify the presence of Trichinella spp. larvae in red foxes, an individual artificial digestion was performed using approximately 30 g of muscle samples. Larvae of Trichinella spp. were detected in one (2.1%) out of the 47 assessed foxes. After a multiplex polymerase chain reaction analysis, T. britovi was molecularly identified as the infecting species. The recognition of T. britovi in a red fox confirms that a sylvatic cycle is present in the North of Portugal and that the local prevalence of Trichinella infection in wildlife must not be ignored due to its underlying zoonotic risks.
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Raposas/parasitologia , Trichinella/isolamento & purificação , Triquinelose/veterinária , Animais , Animais Domésticos , Animais Selvagens , Feminino , Masculino , Músculos/parasitologia , Portugal/epidemiologia , Prevalência , Trichinella/classificação , Trichinella/genética , Triquinelose/epidemiologia , Triquinelose/parasitologiaRESUMO
BACKGROUND: The bacteria Anaplasma platys and Ehrlichia canis and the protozoan Leishmania infantum are vector-borne agents that cause canine vector-borne diseases, some of which are zoonotic. The present survey investigated the prevalence of Anaplasma, Ehrlichia and Leishmania in red foxes (Vulpes vulpes) from Portugal by molecular analysis, in order to evaluate the epidemiological role of these canids as reservoirs of infection. METHODS: Blood and/or bone marrow samples were collected from 78 red foxes obtained in eight districts of northern, central and southern Portugal. Real-time polymerase chain reactions (PCR) amplified a 123 bp fragment of the 16S rRNA gene of Anaplasma spp. and Ehrlichia spp. and a 265 bp fragment of the L. infantum internal transcribed spacer one (ITS1) region of the rRNA operon evaluated by PCR-high resolution melt analysis (PCR-HRM), with sequencing of the DNA products. A phylogenetic analysis was carried out to compare these to other sequences from Anaplasma spp. and Ehrlichia spp. deposited in GenBank. RESULTS: A. platys was detected in 10 (14.5%) and E. canis in two (2.9%) out of 69 foxes; and L. infantum was detected in one (1.3%) of the 78 foxes. The prevalence of A. platys was significantly different from the prevalence of E. canis (p=0.016) and from that of L. infantum (p=0.002). No co-infections were found in any one of the 78 foxes. No statistically significant differences were found between the type of sample (blood and bone marrow), geographic regions (north/centre and south), age (<2 years and ≥2 years) and gender for any one of the agents. CONCLUSIONS: This is the first known report of A. platys in red foxes worldwide, as well as the first molecular evidence of E. canis in foxes from Portugal. The moderate prevalence of A. platys suggests that red foxes may play a role in the epidemiology of infection with this bacterium and serve as a reservoir for domestic dogs.
Assuntos
Anaplasma/isolamento & purificação , Infecções por Anaplasmataceae/veterinária , Ehrlichia canis/isolamento & purificação , Raposas/parasitologia , Leishmania infantum/isolamento & purificação , Leishmaniose Visceral/veterinária , Anaplasma/genética , Infecções por Anaplasmataceae/epidemiologia , Infecções por Anaplasmataceae/parasitologia , Anaplasmose/epidemiologia , Anaplasmose/parasitologia , Animais , Sequência de Bases , DNA de Protozoário/genética , Ehrlichia canis/genética , Ehrlichiose/epidemiologia , Ehrlichiose/parasitologia , Ehrlichiose/veterinária , Leishmania infantum/genética , Leishmaniose Visceral/epidemiologia , Leishmaniose Visceral/parasitologia , Filogenia , Reação em Cadeia da Polimerase , Portugal/epidemiologia , RNA Ribossômico 16S/genéticaRESUMO
Eimeria arloingi infections can cause severe haemorrhagic enteritis in young goat kids, thereby leading to high economic losses in goat industry worldwide. We aimed to isolate a new E. arloingi strain and establish a suitable in vitro culture system for the first merogony. E. arloingi oocysts were collected from naturally infected goat kids in the province of Alentejo, Portugal. For the maintenance of E. arloingi (strain A), kids kept under strict parasite-free conditions were orally infected with 10(3) sporulated oocysts each. Further, a new excystation protocol was successfully established to obtain viable sporozoites for further in vitro development in primary bovine umbilical vein endothelial cells (BUVEC). Overall, E. arloingi first merogony was successfully accomplished in BUVEC leading to macromeront formation (up to 150 µm) and the release of fully developed merozoites I stages. Moreover, host endothelial cell-parasite interactions were investigated in order to determine the extent of modulation carried out by E. arloingi in BUVEC during the first merogony. Gene transcription of adhesion molecules (E-selectin, P-selectin, VCAM-1, ICAM-1) was enhanced in the first hours post-infection (p.i.) in E. arloingi-infected BUVEC. BUVEC activation due to invasion was also shown by increased chemokine (CXCL8, CCL2, CCL5), cytokine (GM-CSF) and COX-2 gene transcription. The new E. arloingi (strain A) will be useful for better comprehension of early host innate immune reactions against this parasite in vitro/in vivo as well as to further our investigations in the complex Eimeria-host endothelial cell interactions.
Assuntos
Eimeria/fisiologia , Animais , Bovinos , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/metabolismo , Células Cultivadas , Quimiocinas , Coccidiose/parasitologia , Eimeria/citologia , Células Endoteliais/parasitologia , Regulação da Expressão Gênica , Humanos , Oocistos/fisiologia , Esporozoítos/imunologiaRESUMO
Coccidiosis caused by Eimeria species is a major form of intestinal infection affecting intensively and semi-intensively reared goats. The province of Alentejo is the main goat-producing area in Portugal. Therefore, all 15 Serpentina goat farms in Alentejo were analyzed regarding the occurrence and diversity of Eimeria species. Fecal samples obtained from 144 animals (52.1% dairy goats, 47.9% pre-pubertal goats) were examined using the modified McMaster technique to determine the number of oocysts per gram of feces. Eimeria spp. oocysts were present in 98.61% of the fecal samples and, overall, nine different Eimeria species were identified. The most prevalent species were E. ninakohlyakimovae (88%) and E. arloingi (85%), followed by E. alijevi (63%) and E. caprovina (63%). The average number of oocysts shed was significantly lower in dairy goats than in pre-adult animals. Astonishingly, no clinical signs of coccidiosis were observed in any of the animals examined, even though they were shedding high numbers of oocysts and were infected with highly pathogenic species. Thus, implementation of routine diagnostic investigation of the occurrence and diversity of caprine Eimeria species may be a useful tool for determination and better understanding of their potential economic impact on goat herds in southern Portugal.
Assuntos
Coccidiose/veterinária , Eimeria , Doenças das Cabras/diagnóstico , Doenças das Cabras/parasitologia , Doenças Parasitárias em Animais/diagnóstico , Animais , Coccidiose/diagnóstico , Feminino , Cabras , PortugalRESUMO
Bovine besnoitiosis, caused by the apicomplexan parasite Besnoitia besnoiti is considered an emergent disease in Europe. This study aimed to determine the prevalence and geographic distribution of B. besnoiti in cattle herds in continental Portugal and to identify potential spatial clustering of infection. A stratified two-stage cross-sectional serological survey was carried out between March 2012 and May 2013 with the five administrative NUTS II regions, Norte, Centro, Lisboa, Alentejo, and Algarve, as the stratification level. Sera from 391 herds in 220 parishes and 83 municipalities were analyzed by a serial testing strategy, with the modified agglutination test (B-MAT) as the first screening assay and the immunofluorescent antibody test (IFAT) as the confirmatory test. Within-herd prevalence of positive herds varied between 0.7 and 72.4% and was ≥10.3% in half of the infected herds. Using a Bayesian approach, the true prevalence of B. besnoiti in cattle herds was determined to be 5.1% (confidence interval (CI), 3.1-7.8%) and the mean within-herd prevalence of positive herds was 33.0% (CI, 20.3-46.0%). The sensitivity and specificity of the B-MAT were estimated to be 96.9% (CI, 93.7-98.8 %) and 99.7% (CI, 99.6-99.8%), whereas those of the IFAT were 89.6% (CI, 86.0-92.5%) and 99.7% (CI, 98.5-99.9%), respectively. Spatial scan statistics analysis identified one spatial cluster covering the majority of the Alentejo region. Seropositive herds were detected for the first time outside Alentejo, in the region Centro and in the northeast of Portugal. Further epidemiological research is needed to identify eco-biological factors, which could explain the geographic clustering of B. besnoiti in Portugal.
Assuntos
Anticorpos Antiprotozoários/sangue , Doenças dos Bovinos/epidemiologia , Coccidiose/veterinária , Sarcocystidae/imunologia , Testes de Aglutinação/veterinária , Animais , Teorema de Bayes , Bovinos , Doenças dos Bovinos/parasitologia , Coccidiose/epidemiologia , Estudos Transversais , Feminino , Técnica Direta de Fluorescência para Anticorpo/veterinária , Masculino , Portugal/epidemiologia , Sarcocystidae/isolamento & purificação , Sensibilidade e Especificidade , Estudos SoroepidemiológicosRESUMO
Besnoitia besnoiti and Toxoplasma gondii are two closely related parasites that interact with the host cell microtubule cytoskeleton during host cell invasion. Here we studied the relationship between the ability of these parasites to invade and to recruit the host cell centrosome and the Golgi apparatus. We observed that T. gondii recruits the host cell centrosome towards the parasitophorous vacuole (PV), whereas B. besnoiti does not. Notably, both parasites recruit the host Golgi apparatus to the PV but its organization is affected in different ways. We also investigated the impact of depleting and over-expressing the host centrosomal protein TBCCD1, involved in centrosome positioning and Golgi apparatus integrity, on the ability of these parasites to invade and replicate. Toxoplasma gondii replication rate decreases in cells over-expressing TBCCD1 but not in TBCCD1-depleted cells; while for B. besnoiti no differences were found. However, B. besnoiti promotes a reorganization of the Golgi ribbon previously fragmented by TBCCD1 depletion. These results suggest that successful establishment of PVs in the host cell requires modulation of the Golgi apparatus which probably involves modifications in microtubule cytoskeleton organization and dynamics. These differences in how T. gondii and B. besnoiti interact with their host cells may indicate different evolutionary paths.
Assuntos
Regulação da Expressão Gênica , Interações Hospedeiro-Parasita , Sarcocystidae/fisiologia , Toxoplasma/fisiologia , Linhagem Celular , Centrossomo/parasitologia , Centrossomo/ultraestrutura , Citoesqueleto , Complexo de Golgi/parasitologia , Complexo de Golgi/ultraestrutura , Humanos , Reprodução , Vacúolos/parasitologia , Vacúolos/ultraestruturaRESUMO
BACKGROUND: Filarioids belonging to the genus Cercopithifilaria (Spirurida: Onchocercidae) have been described in dogs in association with Rhipicephalus sanguineus group ticks, which act as their biological vectors. This study represents the first investigation on Cercopithifilaria spp. in dogs from Portugal. FINDINGS: Dogs (n=102) from the Algarve region (south of Portugal) were sampled by skin snip collection and tissues were left to soak overnight in saline solution. Sediments were observed under a light microscope and the detected microfilariae identified according to their morphology. Twenty-four dogs (23.5%) were found infected with at least one species of Cercopithifilaria, namely C. bainae (9.8%), C. grassii (3.9%) and Cercopithifilaria sp. II sensu Otranto et al., 2013 (13.7%). Results were confirmed by molecular amplification of partial cytochrome c oxidase subunit I and 12S rRNA genes and sequence analysis. Co-infections with more than one Cercopithifilaria species were detected in 3.9% of the animals. CONCLUSIONS: This is the first report of Cercopithifilaria spp. in dogs from Portugal. The estimated level of infection with C. bainae, C. grassii and Cercopithifilaria sp. II suggests that these filarioids are prevalent in the canine population of southern Portugal.
Assuntos
Doenças do Cão/parasitologia , Filariose/veterinária , Filarioidea/isolamento & purificação , Animais , Coinfecção , Doenças do Cão/epidemiologia , Cães , Filariose/epidemiologia , Filariose/parasitologia , Filarioidea/classificação , Portugal/epidemiologia , Prevalência , Pele/parasitologia , Especificidade da EspécieRESUMO
Neosporosis, caused by Neospora caninum, is an important cause of abortion in cattle and of neurological disease in dogs. This study investigated the prevalence and correlates of antibodies to N. caninum in 441 dogs from the five regions of mainland Portugal. A commercial competitive enzyme-linked immunosorbent assay (cELISA) was used and specific antibodies were detected in 35 (7.9%) dogs. Seroprevalence levels were significantly different among some of the studied regions, as well as between stray dogs (13.6%) and hunting dogs (1.7%). The difference between seropositivity in dogs presenting musculoskeletal or neurological signs (21.4%) and that in animals without clinical signs compatible with neosporosis (5.6%) was close to statistical significance. This is the first report on the seroprevalence of N. caninum in dogs in Portugal. Neosporosis should be considered in the differential diagnosis of neurological disorders of dogs.
Assuntos
Anticorpos Antiprotozoários/imunologia , Coccidiose/veterinária , Doenças do Cão/imunologia , Neospora/imunologia , Animais , Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/imunologia , Doenças Assintomáticas , Coccidiose/imunologia , Coccidiose/parasitologia , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Cães/parasitologia , Ensaio de Imunoadsorção Enzimática , Exame Neurológico/veterinária , Portugal/epidemiologia , Prevalência , Estudos Soroepidemiológicos , Avaliação de SintomasRESUMO
Coccidiosis caused by Eimeria species is a major form of intestinal infection affecting intensively and semi-intensively reared goats. The province of Alentejo is the main goat-producing area in Portugal. Therefore, all 15 Serpentina goat farms in Alentejo were analyzed regarding the occurrence and diversity of Eimeria species. Fecal samples obtained from 144 animals (52.1% dairy goats, 47.9% pre-pubertal goats) were examined using the modified McMaster technique to determine the number of oocysts per gram of feces. Eimeria spp. oocysts were present in 98.61% of the fecal samples and, overall, nine different Eimeria species were identified. The most prevalent species were E. ninakohlyakimovae (88%) and E. arloingi (85%), followed by E. alijevi (63%) and E. caprovina (63%). The average number of oocysts shed was significantly lower in dairy goats than in pre-adult animals. Astonishingly, no clinical signs of coccidiosis were observed in any of the animals examined, even though they were shedding high numbers of oocysts and were infected with highly pathogenic species. Thus, implementation of routine diagnostic investigation of the occurrence and diversity of caprine Eimeria species may be a useful tool for determination and better understanding of their potential economic impact on goat herds in southern Portugal.
A coccidiose causada por espécies de Eimeria é a maior infecção intestinal que afeta regimes intensivos e semi-intensivos de caprinos. A região do Alentejo é a mais importante na indústria caprina em Portugal. Assim, todas as 15 explorações de caprinos da raça Serpentina do Alentejo foram analisadas para determinar a frequência e diversidade de espécies de Eimeria presentes. Amostras fecais de 144 animais (52,1% adultas, 47,9% jovens) foram examinadas com a técnica de McMaster modificada para determinar o número de oocistos por grama de fezes. Oocistos de Eimeria spp. estavam presentes em 98,61% das amostras fecais e nove espécies distintas foram identificadas. As espécies mais frequentes foram E. ninakohlyakimovae (88%) e E. arloingi (85%), seguidas por E. alijevi (63%) e E. caprovina (63%). A média do número de oocistos excretados foi significativamente menor em adultas do que em jovens. Surpreendentemente, não foram observados quaisquer sinais clínicos em nenhum dos animais examinados, apesar de eliminarem elevados números de oocistos e de estarem infectados com espécies altamente patogénicas. A prática de diagnósticos de rotina para identificação de espécies de Eimeria caprinas pode ser um importante instrumento para o melhor entendimento do nefasto impacto da doença em explorações de caprinos no Sul de Portugal.
Assuntos
Animais , Feminino , Coccidiose/veterinária , Eimeria , Doenças das Cabras/diagnóstico , Doenças das Cabras/parasitologia , Doenças Parasitárias em Animais/diagnóstico , Coccidiose/diagnóstico , Cabras , PortugalRESUMO
As a novel effector mechanism polymorphonuclear neutrophils (PMN) release neutrophil extracellular traps (NETs), which represent protein-labeled DNA matrices capable of extracellular trapping and killing of invasive pathogens. Here, we demonstrate for the first time NET formation performed by caprine PMN exposed to different stages (sporozoites and oocysts) of the goat apicomplexan protozoan parasite Eimeria arloingi. Scanning electron microscopy as well as fluorescence microscopy of sporozoites- and oocysts-PMN co-cultures revealed a fine network of DNA fibrils partially covering the parasites. Immunofluorescence analyses confirmed the co-localization of histones (H3), neutrophil elastase (NE), and myeloperoxidase (MPO) in extracellular traps released from caprine PMN. In addition, the enzymatic activity of NE was found significantly enhanced in sporozoite-exposed caprine PMN. The treatment of caprine NET structures with deoxyribonuclease (DNase) and the NADPH oxidase inhibitor diphenylene iodondium (DPI) significantly reduced NETosis confirming the classical characteristics of NETs. Caprine NETs efficiently trapped vital sporozoites of E. arloingi since 72% of these stages were immobilized-but not killed-in NET structures. As a consequence, early infection rates were significantly reduced when PMN-pre-exposed sporozoites were allowed to infect adequate host cells. These findings suggest that NETs may play an important role in the early innate host response to E. arloingi infection in goats.
Assuntos
Coccidiose/veterinária , Eimeria/patogenicidade , Doenças das Cabras/imunologia , Imunidade Inata , Neutrófilos/parasitologia , Animais , Células Cultivadas , Coccidiose/imunologia , Técnicas de Cocultura , Espaço Extracelular/imunologia , Doenças das Cabras/parasitologia , Cabras/parasitologia , Histonas/imunologia , Elastase de Leucócito/imunologia , Elastase de Leucócito/metabolismo , Masculino , Microscopia Eletrônica de Varredura , NADPH Oxidases/metabolismo , Ativação de Neutrófilo , Neutrófilos/ultraestrutura , Oocistos , Peroxidase/imunologia , Peroxidase/metabolismo , EsporozoítosRESUMO
Onchocerca lupi, a zoonotic nematode infecting the eyes of carnivores, has been increasingly reported in dogs from Europe and the USA. In order to improve the current status of knowledge on this neglected filarioid, diagnostic imaging tools (i.e., ultrasound scan, computed tomography and magnetic resonance imaging) are herein used to diagnose canine onchocercosis in two dogs, which scored positive for O. lupi microfilariae at the skin snip test and to assess the anatomical location of the nematode within the ocular apparatus. Results indicate that ultrasound tools are useful to address the diagnosis of O. lupi in dogs and to evaluate the localization of nodules or cysts containing the adult nematode.
Assuntos
Doenças do Cão/diagnóstico , Onchocerca/fisiologia , Oncocercose Ocular/veterinária , Animais , Doenças do Cão/diagnóstico por imagem , Cães , Olho/parasitologia , Feminino , Imageamento por Ressonância Magnética/veterinária , Masculino , Oncocercose Ocular/diagnóstico , Oncocercose Ocular/diagnóstico por imagem , Tomografia Computadorizada por Raios X/veterinária , UltrassonografiaRESUMO
Bovine besnoitiosis is caused by the largely unexplored apicomplexan parasite Besnoitia besnoiti. In cows, infection during pregnancy often results in abortion, and chronically infected bulls become infertile. Similar to other apicomplexans B. besnoiti has acquired a largely intracellular lifestyle, but its complete life cycle is still unknown, modes of transmission have not been entirely resolved and the definitive host has not been identified. Outbreaks of bovine besnoitiosis in cattle were described in the 1990s in Portugal and Spain, and later several cases were also detected in France. More cases have been reported recently in hitherto unaffected countries, including Italy, Germany, Switzerland, Hungary and Croatia. To date, there is still no effective pharmaceutical compound available for the treatment of besnoitiosis in cattle, and progress in the identification of novel targets for intervention through pharmacological or immunological means is hampered by the lack of molecular data on the genomic and transcriptomic level. In addition, the lack of an appropriate small animal laboratory model, and wide gaps in our knowledge on the host-parasite interplay during the life cycle of this parasite, renders vaccine and drug development a cost- and labour-intensive undertaking.
Assuntos
Doenças dos Bovinos/parasitologia , Coccidiose/veterinária , Vacinas Protozoárias/imunologia , Sarcocystidae/imunologia , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/prevenção & controle , Coccidiose/diagnóstico , Coccidiose/parasitologia , Coccidiose/prevenção & controle , Feminino , Interações Hospedeiro-Parasita , Masculino , Sarcocystidae/isolamento & purificação , Sarcocystidae/patogenicidadeRESUMO
BACKGROUND: Hepatozoon canis is a protozoan tick-borne pathogen of dogs and wild canids. Hepatozoon spp. have been reported to infect foxes in different continents and recent studies have mostly used the polymerase chain reaction (PCR) for the detection and characterization of the infecting species. Surveying red foxes (Vulpes vulpes) may contribute to better understanding the epidemiology of canine vector-borne diseases, including hepatozoonosis caused by H. canis in domestic dogs. The present study investigated the prevalence of Hepatozoon spp. by means of histopathology and molecular analysis of different tissues in red foxes from different parts of Portugal. METHODS: Blood and tissues including bone marrow, heart, hind leg muscle, jejunum, kidney, liver, lung, popliteal or axillary lymph nodes, spleen and/or tongue were collected from 91 red foxes from eight districts in northern, central and southern Portugal. Tissues were formalin-fixed, paraffin-embedded, cut and stained with hematoxylin and eosin. Polymerase chain reaction (PCR) amplified a ~650 bp fragment of the 18S rRNA gene of Hepatozoon spp. and the DNA products were sequenced. RESULTS: Hepatozoon canis was detected in 68 out of 90 foxes (75.6%) from all the sampled areas by PCR and sequencing. Histopathology revealed H. canis meronts similar in shape to those found in dogs in the bone marrow of 11 (23.4%) and in the spleen of two (4.3%) out of 47 foxes (p = 0.007). All the 11 foxes found positive by histopathology were also positive by PCR of bone marrow and/or blood. Positivity by PCR (83.0%) was significantly higher (p < 0.001) than by histopathological examination (23.4%) in paired bone marrow samples from the same 47 foxes. Sequences of the 18S rRNA gene of H. canis were 98-99% identical to those in GenBank. CONCLUSIONS: Hepatozoon canis was found to be highly prevalent in red fox populations from northern, central and southern Portugal. Detection of the parasite by histopathology was significantly less sensitive than by PCR. Red foxes are a presumptive reservoir of H. canis infection for domestic dogs.