RESUMO
The prevalence of canine Dirofilaria infection in Maio Island (Cape Verde) was analysed by serology, morphological and molecular identification of the parasite species. Blood and sera were collected from 150 dogs and 80 cats aged over 6 months from various localities of the island. DNA was extracted from blood and samples were screened by polymerase chain reaction (PCR) using microfilaria-specific primers. No Dirofilaria immitis was found in dogs while D. repens microfilariae were found in 5.3% of dogs and 6% were positive by PCR. The species identity was confirmed by sequencing of PCR products, which showed almost 100% homology with D. repens European sequences published in GenBank. No difference in Dirofilaria infection was observed between males and females or in dogs with different weights. However, older dogs and those from the western part of Maio Island were more frequently infected. No Dirofilaria was found in cats. This study represents the first evidence of D. repens in Cape Verde (West Africa) and highlights the need for implementing control measures and for a better surveillance of dirofilariosis in Africa.
Assuntos
Dirofilaria repens/isolamento & purificação , Dirofilariose/epidemiologia , Dirofilariose/parasitologia , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Animais , Sangue/parasitologia , Cabo Verde/epidemiologia , Doenças do Gato/epidemiologia , Doenças do Gato/parasitologia , Gatos , DNA de Helmintos/genética , DNA de Helmintos/isolamento & purificação , Dirofilaria repens/genética , Cães , Ilhas/epidemiologia , Reação em Cadeia da Polimerase , Prevalência , Análise de Sequência de DNARESUMO
A survey was carried out to assess the prevalence and the intensity of helminth infections in the roe deer (Capreolus capreolus L.), red deer (Cervus elaphus L.) and fallow deer (Dama dama L.) from 16 hunting areas in western Romania. Overall, 122 deer shot during the hunting season 2013-2015 were examined. Haemonchus contortus, Nematodirus filicollis, Oesophagostomum venulosum and Dictyocalus spp. were found in all the deer species. Trichuris spp. and Dicrocoelium dendriticum were found in the roe deer and red deer and Moniezia expansa in roe deer, only. Overall, the prevalences (<35%) and the mean intensities (<20) were quite low in the abomasums and intestine, but H. contortus in fallow deer (>50%). No >2 or 3 helminth species were found in the abomasum, small and large intestine. Lung helminthes in the roe deer and follow deer accounted for 57.1% and 71.4%, respectively. The only specie with a prevalence >50% was in the red deer. D. dendriticum was found in the liver of roe deer and fallow deer.
RESUMO
Tetracycline treatment of animals or humans infected with filariae that harbor Wolbachia endosymbionts blocks further embryogenesis, and existing microfilariae gradually die. This treatment also kills developing larvae and has a slow-kill effect on adult filariae, all presumably due to elimination of the Wolbachia. Also, Dirofilaria immitis microfilariae in blood collected from dogs up to 25 days after the last dose of doxycycline developed to infective L3 that were normal in appearance and motility in mosquitoes but did not continue to develop or migrate normally after subcutaneous (SC) injection into dogs. The present study was designed to determine whether heartworm microfilariae collected at later times after treatment would regain the ability to continue normal development in a dog. The study also was expected to yield valuable data on the effects of treatment on microfilariae and antigen levels and adult worms. The study was conducted in 16 dogs as two separate replicates at different times. A total of five dogs (two in Replicate A and three in Replicate B) infected either by SC injection of L3 or intravenous transplantation of adult heartworms were given doxycycline orally at 10mg/kg twice daily for 30 days, with three untreated controls. Microfilarial counts in the five treated dogs gradually declined during the 12-13 months after treatment initiation. Two dogs were amicrofilaremic before necropsy and three had 13 or fewer microfilariae/ml. Only one treated dog was negative for heartworm antigen before necropsy. Overall, treated dogs generally had fewer live adult heartworms than controls, and most of their live worms were moribund. All three control dogs remained positive for microfilariae and antigen and had many live worms. L3 from mosquitoes fed on blood collected 73-77 or 161-164 days after initiation of doxycycline treatments were injected SC into five dogs. None of the dogs injected with L3 from mosquitoes fed on blood from doxycycline-treated dogs were ever positive for microfilariae or antigen, and none had worms at necropsy; three control dogs were positive for microfilariae and antigen and had many live worms. These data indicate that doxycycline treatment of microfilaremic dogs gradually reduces numbers of microfilariae and blocks further transmission of heartworms. This latter effect should be highly effective in reducing the rate of selection of heartworms with genes that confer resistance to macrocyclic lactone preventives and microfilaricides. The data also suggest that doxycycline has a slow-kill effect on adult heartworms.
Assuntos
Dirofilaria immitis/efeitos dos fármacos , Dirofilariose/tratamento farmacológico , Doenças do Cão/tratamento farmacológico , Doxiciclina/farmacologia , Doxiciclina/uso terapêutico , Animais , Antígenos de Helmintos/sangue , Culicidae/parasitologia , Dirofilaria immitis/embriologia , Dirofilariose/transmissão , Doenças do Cão/transmissão , Cães , Desenvolvimento Embrionário/efeitos dos fármacos , Microfilárias/efeitos dos fármacosRESUMO
Recently concerns are increasing that dirofilarial nematodes may spread from endemic areas in southern, eastern and central Europe to countries in northern regions of Europe. The increasing number of autochthonous cases of canine Dirofilaria repens infections in Germany indicates that worms of this genus may invade new areas, and climate change may be a key factor in this scenario. Thus analysis of long term development of regional temperature is a pivotal factor in risk analysis related to transmission of these worms. Such information is important for suggestions of counteracting strategies, such as definition of periods of increased transmission risk and, consequently, time slots most suited for preventative measures. In this study, mean daily temperature data from 34 geographical clustered weather stations representing all parts of Germany were analyzed. It is concluded that the increasing trend for average daily temperatures observed in the period from 1984 to 2013 has led to climatic conditions that allow the completion of dirofilarial life cycles in large parts of Germany between May and October. Autochthonous infection with D. repens is already established in some regions and targeted diagnosis and medical prophylaxis is advisable for dogs assumedly exposed during risk of transmission periods. It appears likely that global warming will support further spread of D. repens. Furthermore for the population of dogs the spread of the more pathogenic species D. immitis in hitherto non-endemic Germany is a potential risk if mean temperatures rise to a level suitable for parasite development in the abundant vector mosquitoes during the warmer seasons.
Assuntos
Clima , Dirofilaria immitis/fisiologia , Dirofilaria repens/fisiologia , Dirofilariose/parasitologia , Doenças do Cão/parasitologia , Animais , Análise por Conglomerados , Dirofilariose/epidemiologia , Doenças do Cão/epidemiologia , Cães , Alemanha/epidemiologia , TemperaturaRESUMO
After the repeated demonstration of Dirofilaria repens infections in German dogs, D. repens and Dirofilaria immitis DNA was detected in mosquitoes trapped in 2011, 2012 and 2013 in northeastern and southwestern Germany within the framework of culicid monitoring projects. As temperature is the most important factor dictating the extrinsic development of dirofilarial larvae in the potential vector, data of the German Weather Service (DWD) were analysed for the regions where the positive mosquitoes had been collected. Based on the mean daily temperatures recorded by weather stations most closely to the collection sites of the mosquitoes, it can be concluded that the mosquitoes were trapped in time periods that allowed for the completion of the developmental cycle of the worms in the mosquitoes and a subsequent transmission to a vertebrate host. The results of this study confirm the principal climatic suitability of certain German regions for the establishment of natural dirofilarial transmission cycles. Moreover, the theoretical climatic considerations, together with findings of D. repens infections in German dogs and mosquitoes, strongly suggest that the continuing spread of at least D. repens from its traditional habitats in the Mediterranean has reached southwestern and northeastern Germany.
Assuntos
Culicidae/parasitologia , Dirofilaria immitis/isolamento & purificação , Dirofilaria repens/isolamento & purificação , Dirofilariose/epidemiologia , Temperatura , Animais , Clima , DNA de Helmintos/genética , Dirofilaria immitis/crescimento & desenvolvimento , Dirofilaria repens/crescimento & desenvolvimento , Dirofilariose/transmissão , Doenças do Cão/epidemiologia , Doenças do Cão/transmissão , Cães , Alemanha/epidemiologia , Insetos Vetores/parasitologia , Fatores de TempoRESUMO
Climate change with an increase in average temperature may be responsible for propagation of temperature-dependent vectors and/or vector-associated pathogens in regions that were previously not endemic. The analysis of climate data of Brandenburg state (Germany) had showed that the extrinsic development of Dirofilaria repens, the causative agent of canine subcutaneous dirofilariosis, was possible from the years 2001 to 2012. This finding, associated to the movement of infected dogs and their relocation from endemic European areas to Germany makes possible a rapid endemization of the infection and highlight the risk for human health, being D. repens a frequent cause of zoonotic infections.
Assuntos
Doenças Transmissíveis Emergentes/epidemiologia , Dirofilaria repens/fisiologia , Dirofilariose/epidemiologia , Doenças do Cão/epidemiologia , Doenças Endêmicas , Animais , Clima , Doenças Transmissíveis Emergentes/parasitologia , Dirofilaria repens/efeitos da radiação , Dirofilariose/parasitologia , Doenças do Cão/parasitologia , Cães , Alemanha/epidemiologia , Aquecimento Global , Medição de Risco , TemperaturaRESUMO
Human toxocariasis has been reported to cause a broad spectrum of neurological syndromes, including encephalitis, meningitis and meningo-radiculitis. Nevertheless, cerebral infection by Toxocara may go undiagnosed due to its rarity, elusive symptoms and lack of availability of appropriate testing. We report the case of a 54-year-old man who presented with abdominal pain and paralytic ileus approximately 3 weeks after having eaten raw snails (a folk remedy for peptic ulcer). Three weeks later, marked eosinophilia ensued, associated with mental clouding, nystagmus, diplopia, peripheral limbs ataxia, urinary retention, slackened deep tendon reflexes, arthralgias and myalgias. Cerebrospinal fluid (CSF) examination demonstrated an eosinophilic meningitis, and Toxocara canis cerebral infection was diagnosed by positive serology and by the detection of T. canis DNA in the CSF. The patient made a full recovery following treatment with albendazole and corticosteroids. Physicians should be aware of this rare presentation of toxocariasis, whose diagnosis is, today, facilitated by molecular biology techniques. A history of ingestion of raw snails may alert the clinician to consider the possibility of such an uncommon condition.
Assuntos
Sistema Nervoso Autônomo/fisiopatologia , Encefalite/complicações , Encefalite/patologia , Íleus/etiologia , Toxocara canis/isolamento & purificação , Toxocaríase/complicações , Toxocaríase/patologia , Animais , Anticorpos Anti-Helmínticos/líquido cefalorraquidiano , Líquido Cefalorraquidiano/citologia , DNA de Helmintos/líquido cefalorraquidiano , Encefalite/parasitologia , Doenças Transmitidas por Alimentos/etiologia , Doenças Transmitidas por Alimentos/parasitologia , Doenças Transmitidas por Alimentos/patologia , Humanos , Íleus/parasitologia , Íleus/patologia , Masculino , Pessoa de Meia-Idade , Toxocara canis/genética , Toxocara canis/imunologia , Toxocaríase/parasitologiaRESUMO
Given the spread of Aedes albopictus from northern to southern Italy, and the lack of updated data on Dirofilaria infections, this study was carried out to assess the infection risk for dogs and cats in Apulia region. During a 2-year study, 175 A. albopictus female specimens and samples of blood from 427 dogs (309 privately owned dogs and 118 shelter dogs) and 12 cats were collected. All blood samples were subjected to a modified Knott method, to a test for the detection of circulating Dirofilaria immitis antigen, and to a Dirofilaria species-specific real-time PCR for the simultaneous detection of D. immitis and Dirofilaria repens, targeting on partial cytochrome oxidase subunit 1 and internal transcribed spacer-2, respectively. Two abdomen and one thorax pools from A. albopictus were positive for D. immitis, with minimum infection rates of 1.14 and 0.51, respectively, and a probability of a single positive specimen to be infected of P = 0.6 % (95 % confidence interval (CI) = 0.12-1.73). Out of 439 examined subjects, 22 (5.0 %) tested positive for Dirofilaria spp. in at least one diagnostic test. A specific D. immitis infestation rate of 3.5 % was found among the privately owned dogs, while shelter dogs tested positive only for D. repens with a prevalence of 3.4 %; one cat tested molecularly positive for D. immitis. There was a significantly higher rate of positivity among guard dogs for D. immitis (odds ratio, 6.24, 95 % CI, 1.26-25.28; P < 0.05). The increasing risk of D. immitis infection in southern Italy is supported by the noteworthy positivity of A. albopictus populations and the cat. Our data highlight the usefulness to include filarioid infestation in routine diagnosis.
Assuntos
Aedes/parasitologia , Doenças do Gato/epidemiologia , Dirofilaria/isolamento & purificação , Dirofilariose/epidemiologia , Doenças do Cão/epidemiologia , Animais , Antígenos de Helmintos/sangue , Doenças do Gato/parasitologia , Gatos , DNA de Helmintos/sangue , Dirofilaria/classificação , Dirofilaria/genética , Dirofilaria/imunologia , Dirofilariose/parasitologia , Doenças do Cão/parasitologia , Cães , Feminino , Imunoensaio , Itália/epidemiologia , Prevalência , Reação em Cadeia da Polimerase em Tempo Real , Medição de RiscoRESUMO
In modern taxonomy, DNA barcoding is particularly useful where biometric parameters are difficult to determine or useless owing to the poor quality of samples. These situations are frequent in parasitology. Here, we present an integrated study, based on both DNA barcoding and morphological analysis, on cestodes belonging to the genus Taenia, for which biodiversity is still largely underestimated. In particular, we characterized cestodes from Italian wildcats (Felis silvestris silvestris), free-ranging domestic cats (Felis silvestris catus) and hybrids populations. Adult taeniids were collected by post-mortem examinations of the hosts and morphologically identified as Taenia taeniaeformis. We produced cox1 barcode sequences for all the analysed specimens, and we compared them with reference sequences of individuals belonging to the genus Taenia retrieved from GenBank. In order to evaluate the performance of a DNA barcoding approach to discriminate these parasites, the strength of correlation between species identification based on classical morphology and the molecular divergence of cox1 sequences was measured. Our study provides clear evidence that DNA barcoding is highly efficient to reveal the presence of cryptic lineages within already-described taeniid species. Indeed, we detected three well-defined molecular lineages within the whole panel of specimens morphologically identified as T. taeniaeformis. Two of these molecular groups were already identified by other authors and should be ranked at species level. The third molecular group encompasses only samples collected in Italy during this study, and it represents a third candidate species, still morphologically undescribed.
Assuntos
Código de Barras de DNA Taxonômico/métodos , Taenia/classificação , Taenia/genética , Animais , Doenças do Gato/parasitologia , Gatos , Ciclo-Oxigenase 1/genética , DNA Mitocondrial/química , DNA Mitocondrial/genética , Itália , Dados de Sequência Molecular , Análise de Sequência de DNA , Taenia/isolamento & purificação , Teníase/parasitologia , Teníase/veterináriaRESUMO
Cats are considered a susceptible host for Dirofilaria immitis; however, increased host resistance is reflected by relatively low adult worm burdens in natural and experimental infections; the prolonged prepatent period (8 months); the low level and short duration of microfilaremia; and the short life span of adult worms (2-3 years). From April to September 2006, 212 cats and 608 dogs, all exposed for at least one transmission season, were screened for D. immitis infection in a multi-center study in the Po River Valley in northern Italy. Cats were initially evaluated by antibody testing; positive subjects were followed up by antigen testing and echocardiography (and necropsy if death occurred). The prevalence in dogs was 29% by a modified Knott test and antigen testing compared with a prevalence of 4.7% in cats by an antibody test; six of these infections (2.8%) were confirmed by the follow-up evaluations. This field study demonstrated that the prevalence of heartworm infection in cats in this area is within the expected limits of 9-18% of the prevalence in dogs. Antibody testing likely underestimates the real prevalence of D. immitis infection in cats. These results also emphasize the importance of preventive treatment in cats.
Assuntos
Doenças do Gato/epidemiologia , Dirofilaria immitis/imunologia , Dirofilariose/epidemiologia , Doenças do Cão/epidemiologia , Animais , Antígenos de Helmintos/imunologia , Doenças do Gato/diagnóstico por imagem , Doenças do Gato/parasitologia , Gatos , Dirofilaria immitis/patogenicidade , Dirofilariose/diagnóstico por imagem , Dirofilariose/parasitologia , Doenças do Cão/parasitologia , Cães , Ecocardiografia/veterinária , Feminino , Itália/epidemiologia , Expectativa de Vida , Masculino , PrevalênciaRESUMO
Geospatial tools (e.g., geographical information systems, remote sensing, global positioning systems, and virtual globes) are very useful for the simultaneous visualization of health data with environmental data, which holds promise to understand environmental-health linkages and to generate new hypotheses to be tested in future research. Current epidemiological studies clearly show that the distribution patterns of vector-borne infections are changing; for example, in Europe, heartworm infection and subcutaneous dirofilariosis are spreading throughout areas that previously had little to no incidence of heartworm. In view of the changes of the distribution patterns of Dirofilaria immitis and Dirofilaria repens, geospatial tools are now more useful for mapping (including territorial sampling), monitoring, ecological analysis, risk assessment, forecasting (including the choose of the timing of treatment), early warning, and surveillance of both heartworm and subcutaneous dirofilariosis. All these issues have control of these infections as the ultimate goal.
Assuntos
Doenças do Gato/prevenção & controle , Dirofilariose/prevenção & controle , Doenças do Cão/prevenção & controle , Sistemas de Informação Geográfica/normas , Animais , Doenças do Gato/epidemiologia , Doenças do Gato/parasitologia , Gatos , Coleta de Dados , Dirofilaria immitis/patogenicidade , Dirofilaria repens/patogenicidade , Dirofilariose/epidemiologia , Dirofilariose/parasitologia , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Cães , Ecossistema , Europa (Continente)/epidemiologia , Previsões , Vigilância da População , Medição de RiscoRESUMO
The antifilarial effects of tetracycline drugs were first demonstrated when they were found to be highly effective against L(3) and L(4) of Brugia pahangi and Litomosoides sigmodontis in rodent models. Tetracyclines are also now known to have activity against microfilariae and adult Dirofilaria immitis, but assessment of their activity against larval and juvenile heartworms has not been reported previously. This study assessed the effects of doxycycline administered orally at 10mg/kg twice daily for 30-day periods at selected times during the early part of the life cycle of D. immitis in dogs with dual infections of D. immitis and B. pahangi. Twenty beagles were randomly allocated by weight to four groups of five dogs each. On Day 0, each dog was given 50 D. immitis L(3) and 200 B. pahangi L(3) by SC injection. Dogs received doxycycline on Days 0-29 (Group 1); Days 40-69 (Group 2); or Days 65-94 (Group 3). Group 4 served as untreated controls. Blood samples were collected for microfilariae counting and antigen testing. Necropsy for collection of adult heartworms and selected tissues were performed Days 218-222. Heartworms recovered were examined by immunohistology, conventional microscopy/transmission electron microscopy, and molecular biology techniques. No live heartworms were recovered from dogs in Group 1; dogs in Group 2 had 0 to 2 live worms (98.4% efficacy), and dogs in Group 3 had 0-36 live worms (69.6% efficacy). All control dogs had live adult heartworms (25-41). The live worms recovered from dogs in Groups 2 and 3 were less developed and smaller that worms from control dogs. Microfilariae were not detected in any dogs in Groups 1 and 2; one dog in Group 3 had 1 microfilariae/ml at necropsy. All control dogs had microfilariae at necropsy. One dog in Group 1 was antigen positive at one sampling (Day 166). One dog in Group 2 was antigen positive Days 196 and 218-222 and three dogs in Group 3 were antigen positive at one or more samplings All five control dogs were antigen positive at all three sampling times. These findings suggest that doxycycline at 10mg/kg orally twice daily for 30 days has efficacy against migrating tissue-phase larvae and juvenile worms and will delay or restrict microfilarial production.
Assuntos
Antibacterianos/uso terapêutico , Brugia pahangi/efeitos dos fármacos , Dirofilariose/tratamento farmacológico , Doenças do Cão/tratamento farmacológico , Doxiciclina/uso terapêutico , Filariose/veterinária , Administração Oral , Animais , Antibacterianos/administração & dosagem , Antibacterianos/farmacologia , Antígenos de Helmintos/sangue , Brugia pahangi/patogenicidade , Dirofilaria immitis/efeitos dos fármacos , Dirofilaria immitis/patogenicidade , Dirofilariose/complicações , Dirofilariose/parasitologia , Doenças do Cão/parasitologia , Cães , Doxiciclina/administração & dosagem , Doxiciclina/farmacologia , Feminino , Filariose/complicações , Filariose/tratamento farmacológico , Larva/efeitos dos fármacos , Larva/patogenicidade , Masculino , Microfilárias/efeitos dos fármacos , Microfilárias/patogenicidade , Distribuição Aleatória , Fatores de TempoRESUMO
Dirofilaria immitis and Dirofilaria repens are the most common species of filarial nematodes described in the dogs with increasing spread into new geographical areas. The diagnosis of canine dirofilariosis is usually based upon the microscopical detection and identification of circulating microfilariae together with ELISA detection of serum circulating heartworm antigens or antibodies. The identification of the parasite species using the traditional approaches sometimes can be difficult and can lead to misdiagnosis especially on samples from areas where both Dirofilaria are present. In this paper we report a new molecular method based on single-step multiplex PCR to detect and differentiate simultaneously and unequivocally D. immitis and D. repens on DNA extracted from canine peripheral blood. The amplification was performed using a set of primers designed on a portion of the small subunit ribosomal RNA gene of the mitochondrion (12S rDNA). The single-step multiplex PCR here described ensured high (4 mf/ml) sensitivity and specificity with reduced cost and time saving. The multiplex PCR assay represents an additional tool for epidemiological studies and routine disease assessment in areas co-endemic for the two Dirofilaria species.
Assuntos
Dirofilaria immitis/isolamento & purificação , Dirofilariose/parasitologia , Doenças do Cão/parasitologia , Reação em Cadeia da Polimerase/veterinária , Animais , DNA de Helmintos/química , DNA de Helmintos/genética , Dirofilaria immitis/genética , Dirofilariose/diagnóstico , Doenças do Cão/diagnóstico , Cães , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico/química , RNA Ribossômico/genética , Sensibilidade e Especificidade , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
A five-year-old, entire female mixed-breed dog was presented with corneal oedema and episcleral hyperaemia in the left eye. The ophthalmological examination revealed the presence of a free-swimming nematode in the anterior chamber. Circulating microfilariae were not observed by a modified Knott test nor were adult antigens detected in serum by a commercial ELISA. The parasite was surgically removed from the dog's eye, but its anterior end was damaged during the surgery. Based on the morphology of the posterior end, the nematode was preliminarily identified as a male Dirofilaria immitis. The species identification was confirmed by PCR amplification and sequencing of the mitochondrial coxI and 12S rDNA genes, using a DNA barcoding approach. Although other cases of ocular dirofilariosis by D. immitis have been previously recorded in Australia and the United States, the case reported herein is the first in a dog from Europe.
Assuntos
Dirofilaria immitis , Dirofilariose/diagnóstico , Doenças do Cão/diagnóstico , Infecções Oculares Parasitárias/veterinária , Animais , DNA de Helmintos/análise , Dirofilaria immitis/isolamento & purificação , Dirofilariose/cirurgia , Doenças do Cão/cirurgia , Cães , Europa (Continente)/epidemiologia , Infecções Oculares Parasitárias/diagnóstico , Infecções Oculares Parasitárias/cirurgia , Feminino , Masculino , Reação em Cadeia da Polimerase/veterináriaRESUMO
Echinococcus multilocularis is characterised by a wide geographical distribution, encompassing three continents (North America, Asia and Europe) yet very low genetic variability is documented. Recently, this parasite has been detected in red foxes (Vulpes vulpes) circulating in an Alpine region of Italy, close to Austria. This finding raised the question as to whether an autochthonous cycle exists in Italy or whether the infected foxes originated from the neighbouring regions of Austria. Studies have shown that multi-locus microsatellite analysis can identify genomic regions carrying mutations that result in a local adaptation. We used a tandem repeated multi-locus microsatellite (EmsB) to evaluate the genetic differences amongst adult worms of E. multilocularis collected in Italy, worms from neighbouring Austria and from other European and extra-European countries. Fluorescent PCR was performed on a panel of E. multilocularis samples to assess intra-specific polymorphism. The analysis revealed four closed genotypes for Italian samples of E. multilocularis which were unique compared with the other 25 genotypes from Europe and the five genotypes from Alaska. An analysis in the Alpine watershed, comparing Italian adult worms with those from neighbouring areas in Austria, showed a unique cluster for Italian samples. This result supports the hypothesis of the presence of an autochthonous cycle of E. multilocularis in Italy. EmsB can be useful for 'tracking' the source of infection of this zoonotic parasite and developing appropriate measures for preventing or reducing the risk of human alveolar echinococcosis.
Assuntos
Equinococose/epidemiologia , Echinococcus multilocularis/genética , Sequências de Repetição em Tandem/genética , Animais , Análise por Conglomerados , DNA de Helmintos/genética , Equinococose/veterinária , Echinococcus multilocularis/isolamento & purificação , Echinococcus multilocularis/parasitologia , Europa (Continente)/epidemiologia , Raposas/parasitologia , Genótipo , Humanos , Itália/epidemiologia , Repetições de Microssatélites , Epidemiologia Molecular , Reação em Cadeia da Polimerase/métodos , Polimorfismo Genético/genética , Prevalência , Reprodutibilidade dos TestesRESUMO
Antigens of both Dirofilaria immitis and Wolbachia symbiont bacteria are implicated in the inflammatory pathology of heartworm infection. The aim of the present study was to compare the stimulatory capacity of in vitro cultures of vascular endothelial cells by the adult somatic antigens of D. immitis (DiSA) and the recombinant form of the Wolbachia surface protein (rWSP), during the first 24h of stimulation. Our results indicate a different stimulatory activity of the two antigens. Both the DiSA and rWSP stimulate the production of the enzymes responsible of the arachidonic acid metabolism, cyclooxygenase-2, 5-lipoxygenase (5-LO), and leukotriene B4. Only DiSA stimulates the production of prostaglandin E2. Related to the adhesion molecules, the DiSA stimulates the expression of intercellular adhesion molecule-1 (ICAM-1) and platelet endothelial cell adhesion molecule-1 (PECAM-1), whereas rWSP stimulates ICAM-1, PECAM-1, and vascular cell adhesion molecule-1 (VCAM-1). Expression of E-cadherin and vascular endothelial growth factor also were stimulated by rWSP. Neither of the two antigens altered the basic physiological mechanisms of endothelial cells, such as cell proliferation, cell cycle, or apoptosis. The biological and pathological significance of these finding are discussed.
Assuntos
Antígenos de Helmintos/imunologia , Dirofilaria immitis/imunologia , Endotélio Vascular/citologia , Proteínas de Membrana/imunologia , Wolbachia/imunologia , Animais , Células Cultivadas , Células Endoteliais/imunologia , Células Endoteliais/metabolismo , Endotélio Vascular/imunologia , Endotélio Vascular/metabolismo , Humanos , Inflamação/imunologia , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/imunologia , Molécula 1 de Adesão Intercelular/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/genética , Molécula-1 de Adesão Celular Endotelial a Plaquetas/imunologia , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Molécula 1 de Adesão de Célula Vascular/genética , Molécula 1 de Adesão de Célula Vascular/imunologia , Molécula 1 de Adesão de Célula Vascular/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/imunologia , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
A safer, more effective adulticidal treatment and a safe method for reducing microfilaremia and breaking transmission of heartworm disease early in the treatment are needed. The present study evaluated efficacy of ivermectin (IVM) and doxycycline (DOXY) alone or together (with or without melarsomine [MEL]) in dogs with induced adult heartworm infection and assessed the ability of microfilariae from DOXY-treated dogs to develop to L3 in Aedes aegypti mosquitoes and subsequently to become reproductive adults in dogs. Thirty beagles were each infected with 16 adult heartworms by intravenous transplantation. Six weeks later, dogs were ranked by microfilarial count and randomly allocated to 6 groups of 5 dogs each. Beginning on Day 0, Group 1 received IVM (6 mcg/kg) weekly for 36 weeks. Group 2 received DOXY (10 mcg/(kgday)) orally Weeks 1-6, 10-11, 16-17, 22-25, and 28-33. Groups 3 and 5 received IVM and DOXY according to doses and schedules used for Groups 1 and 2. At Week 24, Groups 3 and 4 received an intramuscular injection of MEL (2.5 mg/kg), followed 1 month later by two injections 24h apart. Group 6 was not treated. Blood samples were collected for periodic microfilaria counts and antigen (Ag) testing (and later immunologic evaluation and molecular biology procedures). Radiographic and physical examinations, hematology/clinical chemistry testing, and urinalysis were done before infection, before Day 0, and periodically during the treatment period. At 36 weeks, the dogs were euthanized and necropsied for worm recovery, collection of lung, liver, kidney, and spleen samples for examination by immunohistochemistry and conventional histological methods. All dogs treated with IVM + DOXY (with or without MEL) were amicrofilaremic after Week 9. Microfilarial counts gradually decreased in dogs treated with IVM or DOXY, but most had a few microfilariae at necropsy. Microfilarial counts for dogs treated only with MEL were similar to those for controls. Antigen test scores gradually decreased with IVM + DOXY (with or without MEL) and after MEL. Antigen scores for IVM or DOXY alone were similar to controls throughout the study. Reduction of adult worms was 20.3% for IVM, 8.7% for DOXY, 92.8% for IVM + DOXY + MEL, 100% for MEL, and 78.3% for IVM + DOXY. Mosquitoes that fed on blood from DOXY-treated dogs had L3 normal in appearance but were not infective for dogs. Preliminary observations suggest that administration of DOXY+IVM for several months prior to (or without) MEL will eliminate adult HW with less potential for severe thromboembolism than MEL alone.
Assuntos
Arsenicais/uso terapêutico , Dirofilaria immitis/microbiologia , Dirofilariose/tratamento farmacológico , Doxiciclina/uso terapêutico , Filaricidas/uso terapêutico , Ivermectina/uso terapêutico , Triazinas/uso terapêutico , Aedes/microbiologia , Animais , Antibacterianos/efeitos adversos , Antibacterianos/uso terapêutico , Antiparasitários/efeitos adversos , Antiparasitários/uso terapêutico , Arsenicais/efeitos adversos , Dirofilaria immitis/efeitos dos fármacos , Doenças do Cão/tratamento farmacológico , Cães , Relação Dose-Resposta a Droga , Doxiciclina/efeitos adversos , Feminino , Filaricidas/efeitos adversos , Ivermectina/efeitos adversos , Masculino , Microfilárias , Distribuição Aleatória , Tromboembolia/induzido quimicamente , Tromboembolia/prevenção & controle , Tromboembolia/veterinária , Fatores de Tempo , Resultado do Tratamento , Triazinas/efeitos adversos , Wolbachia/efeitos dos fármacosRESUMO
Since the definitive identification in 1995 of the bacterial endosymbiont Wolbachia that resides in different tissues of the filarial worm Dirofilaria immitis, there has been increasing interest to understand whether and what role it plays in the pathogenesis of and immune response to heartworm infection. The present study evaluated the effects of treatments on lung pathology in 20 beagle dogs experimentally infected with D. immitis. Dogs in Group 1 were treated with doxycycline (10 mg/kg/day) orally from weeks 0-6, 10-12, 16-18, 22-26, and 28-34. Dogs in Group 2 served as infected, non-treated controls. Dogs in Group 3 were given doxycycline as described for Group 1 combined with weekly oral doses of ivermectin (6 mcg/kg) for 34 weeks and intramuscular (IM) melarsomine (2.5 mg/kg) at week 24, followed by two additional melarsomine injections 24h apart 1 month later. Group 4 received only melarsomine as described for Group 3. Lung lesion criteria, scored by two independent blinded pathologists, included perivascular inflammation and endothelial proliferation. Doxycycline treatment alone had no effect on lesion scores, whereas the combination of doxycycline and ivermectin resulted in less severe perivascular inflammation. All lungs were evaluated for positive immunostaining for the Wolbachia surface protein (WSP). Control dogs showed numerous thrombi, intense perivascular and interstitial inflammation and, occasionally, positive staining for WSP. Interestingly, dogs receiving doxycycline/ivermectin/melarsomine showed significantly less severe arterial lesions and the virtual absence of thrombi.
Assuntos
Antibacterianos/uso terapêutico , Dirofilaria immitis/microbiologia , Dirofilariose/patologia , Doenças do Cão/patologia , Filaricidas/uso terapêutico , Pulmão/patologia , Wolbachia/imunologia , Animais , Arsenicais/uso terapêutico , Proteínas da Membrana Bacteriana Externa/imunologia , Dirofilaria immitis/patogenicidade , Dirofilariose/imunologia , Dirofilariose/microbiologia , Dirofilariose/parasitologia , Doenças do Cão/imunologia , Doenças do Cão/microbiologia , Doenças do Cão/parasitologia , Cães , Doxiciclina/uso terapêutico , Feminino , Imuno-Histoquímica/veterinária , Ivermectina/uso terapêutico , Pulmão/parasitologia , Masculino , Índice de Gravidade de Doença , Triazinas/uso terapêutico , Wolbachia/efeitos dos fármacosRESUMO
Clinical manifestations of heartworm disease in cats are variable; most cats seem to tolerate the infection well for extended periods. Heartworm-infected cats may undergo spontaneous self-cure due to the natural death of parasites without any symptomatology, or they may suddenly show dramatic and acute symptoms. Sudden death in apparently healthy cats is not a rare event. Thoracic radiographs are important tool for the diagnosis of cardiopulmonary disease. However, thoracic abnormalities are often absent or transient and highly variable in heartworm-infected cats. Findings, such as enlargement of the peripheral branches of the pulmonary arteries, with a varying degree of pulmonary parenchymal disease and hyperinflation, are the most typical features consistent with infection. A field study was performed for cats referred to the Veterinary Hospital Città di Pavia from January 1998 to December 2001 for routine health examinations and procedures to evaluate the clinical evolution and radiographic findings of feline heartworm infection. Thirty-four asymptomatic cats diagnosed with feline heartworm infection by antibody and antigen tests together with an echocardiogram that allowed worm visualization were included in the follow-up study. Cats were routinely examined every 3 months from the time of heartworm diagnosis until the outcome (self-cure or death). Self-cure was defined as no positive serology for heartworm antigens and no visualization of worms by echocardiography. A final examination for antibodies was carried after 12 months as a final confirmation of self-cure. Twenty-eight cats (82.4%) self-cured; including 21 that showed no clinical signs of infection throughout the study. Six cats died. The most common clinical features observed were acute respiratory symptoms and sudden death. Infection lasted over 3 years in the majority of the cats enrolled in the study. Thoracic radiograph appearance was variable, and the most commonly observed findings were focal and diffuse pulmonary parenchymal abnormalities.