RESUMO
Vascular endothelial growth factor (VEGF)-A is differentially spliced to give two functionally different isoform families; pro-angiogenic, pro-permeability VEGF-Axxx and anti-angiogenic, anti-permeability VEGF-Axxxb. VEGF-A splicing is dysregulated in several pathologies, including cancer, diabetes, and peripheral arterial disease. The bichromatic VEGF-A splicing-sensitive fluorescent reporter harboured in a transgenic mouse is a novel approach to investigate the splicing patterns of VEGF-A in vivo. We generated a transgenic mouse harbouring a splicing-sensitive fluorescent reporter designed to mimic VEGF-A terminal exon splicing (VEGF8ab) by insertion into the ROSA26 genomic locus. dsRED expression denotes proximal splice site selection (VEGF-Axxx) and eGFP expression denotes distal splice site selection (VEGF-Axxxb). We investigated the tissue-specific expression patterns in the eye, skeletal muscle, cardiac muscle, kidney, and pancreas, and determined whether the splicing pattern could be manipulated in the same manner as endogenous VEGF-A by treatment with the SRPK1 inhibitor SPHINX 31. We confirmed expression of both dsRED and eGFP in the eye, skeletal muscle, cardiac muscle, kidney, and pancreas, with the highest expression of both fluorescent proteins observed in the exocrine pancreas. The ratio of dsRED and eGFP matched that of endogenous VEGF-Axxx and VEGF-Axxxb. Treatment of the VEGF8ab mice with SPHINX 31 increased the mRNA and protein eGFP/dsRED ratio in the exocrine pancreas, mimicking endogenous VEGF-A splicing. The VEGF-A exon 8 splicing-sensitive fluorescent reporter mouse is a novel tool to assess splicing regulation in the individual cell-types and tissues, which provides a useful screening process for potentially therapeutic splicing regulatory compounds in vivo.
Assuntos
Processamento Alternativo , Animais Geneticamente Modificados , Genes Reporter , Proteínas de Fluorescência Verde/genética , Proteínas Luminescentes/genética , Fator A de Crescimento do Endotélio Vascular/genética , Animais , Éxons , Olho/metabolismo , Loci Gênicos , Proteínas de Fluorescência Verde/metabolismo , Íntrons , Rim/metabolismo , Proteínas Luminescentes/metabolismo , Camundongos , Músculo Esquelético/metabolismo , Miocárdio/metabolismo , Pâncreas/metabolismo , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Inibidores de Proteínas Quinases/farmacologia , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Neospora caninum is a commonly diagnosed cause of reproductive losses in farmed ruminants worldwide. This study examined 495 and 308 samples (brain, heart and placenta) which were collected from 455 and 119 aborted cattle and sheep fetuses, respectively. DNA was extracted and a nested Neospora ITS1 PCR was performed on all samples. The results showed that for bovine fetuses 79/449 brain [17.6% (14.2-21.4)], 7/25 heart [28.0% (12.1-49.4)] and 5/21 placenta [23.8% (8.2-47.2)] were PCR positive for the presence of Neospora DNA. Overall 82/455 [18.0% (14.6-21.7)] of the bovine fetuses tested positive for the presence of N. caninum DNA in at least one sample. None (0/308) of the ovine fetal samples tested positive for the presence of Neospora DNA in any of the tissues tested. The results show that N. caninum was associated with fetal losses in cattle (distributed across South-West Scotland), compared to sheep in the same geographical areas where no parasite DNA was found. Neospora is well distributed amongst cattle in South-West Scotland and is the potential cause of serious economic losses to the Scottish cattle farming community; however, it does not appear to be a problem amongst the Scottish sheep flocks.
Assuntos
Aborto Animal/parasitologia , Doenças dos Bovinos/parasitologia , DNA de Protozoário/isolamento & purificação , Neospora/isolamento & purificação , Doenças dos Ovinos/parasitologia , Feto Abortado/parasitologia , Animais , Encéfalo/parasitologia , Bovinos , DNA Intergênico/isolamento & purificação , Fazendas/estatística & dados numéricos , Feminino , Coração/parasitologia , Placenta/parasitologia , Gravidez , Complicações Parasitárias na Gravidez/parasitologia , OvinosRESUMO
Neck samples from 54 badgers and 32 tongue samples of the same badgers (Meles meles), collected in the Lothians and Borders regions of Scotland, were tested using polymerase chain reactions (PCRs) directed against the 18S ribosomal DNA and the internal transcribed spacer (ITS1) region of protozoan parasites of the family Sarcocystidae. Positive results were obtained from 36/54 (67%) neck and 24/32 (75%) tongue samples using an 18S rDNA PCR. A 468 base pair consensus sequence that was generated from the 18S rDNA PCR amplicons (KX229728) showed 100% identity to Sarcocystis lutrae. The ITS1 PCR results revealed that 12/20 (60%) neck and 10/20 (50%) tongue samples were positive for Sarcocystidae DNA. A 1074 bp consensus sequence was generated from the ITS1 PCR amplicons (KX431307) and showed 100% identity to S. lutrae. Multiple sequence alignments and phylogenetic analysis support the finding that the rDNA found in badgers is identical to that of S. lutrae. This parasite has not been previously reported in badgers or in the UK. Sarcocystis lutrae has previously only been detected in tongue, skeletal muscle and diaphragm samples of the Eurasian otter (Lutra lutra) in Norway and potentially in the Arctic fox (Vulpes lagopus).
Assuntos
DNA de Protozoário/genética , Mustelidae/parasitologia , Sarcocystis/genética , Sarcocystis/isolamento & purificação , Sarcocistose/veterinária , Animais , DNA Ribossômico , DNA Espaçador Ribossômico/genética , Técnicas de Diagnóstico Molecular , Filogenia , Reação em Cadeia da Polimerase , Sarcocystis/classificação , Sarcocistose/diagnóstico , Sarcocistose/epidemiologia , Sarcocistose/parasitologia , Escócia/epidemiologia , Análise de Sequência de DNARESUMO
Toxoplasma gondii is a zoonotic pathogen defined by three main clonal lineages (types I, II, III), of which type II is most common in Europe. Very few data exist on the prevalence and genotypes of T. gondii in the UK. Wildlife can act as sentinel species for T. gondii genotypes present in the environment, which may subsequently be transmitted to livestock and humans. DNA was extracted from tissue samples of wild British carnivores, including 99 ferrets, 83 red foxes, 70 polecats, 65 mink, 64 badgers and 9 stoats. Parasite DNA was detected using a nested ITS1 PCR specific for T. gondii, PCR positive samples were subsequently genotyped using five PCR-RFLP markers. Toxoplasma gondii DNA was detected within all these mammal species and prevalence varied from 6·0 to 44·4% depending on the host. PCR-RFLP genotyping identified type II as the predominant lineage, but type III and type I alleles were also identified. No atypical or mixed genotypes were identified within these animals. This study demonstrates the presence of alleles for all three clonal lineages with potential for transmission to cats and livestock. This is the first DNA-based study of T. gondii prevalence and genotypes across a broad range of wild British carnivores.
Assuntos
Carnívoros , Toxoplasma/genética , Toxoplasmose Animal/parasitologia , Animais , Sequência de Bases , DNA Intergênico/genética , DNA de Protozoário/genética , Regulação da Expressão Gênica , Variação Genética , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de Restrição , Alinhamento de Sequência , Especificidade da Espécie , Toxoplasma/classificação , Toxoplasmose Animal/epidemiologia , Reino UnidoRESUMO
Balb/c mice were inoculated intraperitoneally (i.p.) with either 5 x 10(6) live virulent (group 1) or 5 x 10(6) live attenuated (group 2) tachyzoites, or Vero cells (group 3). Animals were killed at 0, 14, 28 and 42 days post-inoculation (p.i.), with the remaining mice receiving a lethal challenge on day 48 p.i. Serum, spleen and brain samples were collected post-mortem to examine humoral and cell-mediated immune responses as well as pathological lesions and to quantify parasite loads. On day 14 p.i. group 2 (attenuated) demonstrated statistically significant (P < 0.001) lower levels of mean morbidity and weight loss, while also showing significantly (P = 0.01) higher levels of splenocyte proliferation and IFN-gamma production (P = 0.003), compared to group 1 (virulent). Histology of brain samples showed milder lesions and a lower incidence of positive immunohistochemistry, demonstrating tachyzoites and tissue cysts, and statistically significant (P = 0.03) lower mean burdens of parasite DNA in group 2 (attenuated) compared to group 1 (virulent). All mice in group 2 were protected following challenge on day 48 p.i. whereas naïve control mice succumbed to the challenge. No mice from group 1 (virulent) survived beyond day 24 p.i. so they were not included in the challenge.
Assuntos
Coccidiose/imunologia , Coccidiose/prevenção & controle , Neospora/imunologia , Animais , Anticorpos Antiprotozoários/sangue , Peso Corporal , Encéfalo/imunologia , Encéfalo/parasitologia , Encéfalo/patologia , Coccidiose/parasitologia , Feminino , Interferon gama/metabolismo , Leucócitos Mononucleares/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Soro/imunologia , Soro/parasitologia , Índice de Gravidade de Doença , Baço/imunologia , Baço/parasitologia , Análise de SobrevidaRESUMO
A 33-year-old multiparous term parturient requested an epidural for labour analgesia. An accidental dural puncture was noted at the time of epidural needle insertion and an intrathecal catheter was placed. The intrathecal catheter was removed 10 h later and was noted to be intact. The following day, there was a significant leak of clear fluid from the catheter insertion site. Cerebrospinal fluid was detected using beta-trace protein testing. The patient remained asymptomatic and we deduced the cause could be a cutaneous fistula. Similar published reports have suggested the causes of similar presentations could include leakage of interstitial oedema, local anaesthetic or cerebrospinal fluid. It is important to consider the diagnosis of clear fluid leakage following an accidental dural puncture and the management of a cerebrospinal fluid-cutaneous fistula in the absence of neurological symptoms.
RESUMO
PURPOSE: Toxoplasma gondii is a zoonotic parasite capable of infecting a wide range of hosts. Free-range chickens are important sentinels in the epidemiology of this parasite as they feed from the ground and are likely to ingest oocysts shed in the faeces of infected cats. Atypical strains of T. gondii are known to dominate in South America where they are associated with more severe disease in humans, yet relatively little is known about the strains circulating in neighbouring Caribbean islands. METHODS: In this study, hearts and brains were collected from free-range chickens in Antigua and Barbuda (n = 45), Dominica (n = 76) and Trinidad (n = 41), and DNA was extracted for nested ITS1 PCR and PCR-RFLP. Sera were collected and screened for antibodies using the modified agglutination test (MAT). RESULTS: Antibodies to T. gondii were detected in 20.5, 38.2 and 17.1% of chickens in Antigua and Barbuda, Dominica and Trinidad, respectively. Toxoplasma gondii DNA was also detected by PCR in 24.4, 17.1 and 17.1% of chickens, respectively, giving an overall prevalence of 31.1, 42.1, and 29.3% for each of the 3 island nations. Results of PCR-RFLP revealed 2 new atypical genotypes (designated ToxoDB #281 and #282) and one Type III (ToxoDB #2) in chickens from Antigua. Partial genotyping of a further 8 isolates (7 from Antigua and one from Trinidad) revealed different allele-types at five or more markers for 7 of the isolates, suggesting atypical genotypes. CONCLUSIONS: This is the first study to report the prevalence of T. gondii in free-range chickens in Antigua and Barbuda, Dominica and Trinidad and Tobago. It is also the first to report the presence of atypical genotypes in Antigua and Barbuda and Trinidad and Tobago.
Assuntos
Galinhas/parasitologia , Variação Genética , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/parasitologia , Toxoplasma/genética , Toxoplasmose Animal/epidemiologia , Animais , Anticorpos Antiprotozoários/sangue , Encéfalo/parasitologia , DNA de Protozoário/genética , Genótipo , Coração/parasitologia , Prevalência , Índias Ocidentais/epidemiologiaRESUMO
In cattle, antibodies to Toxoplasma gondii infection are frequently detected, but evidence for the presence of T. gondii tissue cysts in cattle is limited. To study the concordance between the presence of anti-T. gondii IgG and viable tissue cysts of T. gondii in cattle, serum, liver and diaphragm samples of 167 veal calves and 235 adult cattle were collected in Italy, the Netherlands, Romania and the United Kingdom. Serum samples were tested for anti-T. gondii IgG by the modified agglutination test and p30 immunoblot. Samples from liver were analyzed by mouse bioassay and PCR after trypsin digestion. In addition, all diaphragms of cattle that had tested T. gondii-positive (either in bioassay, by PCR on trypsin-digested liver or serologically by MAT) and a selection of diaphragms from cattle that had tested negative were analyzed by magnetic capture quantitative PCR (MC-PCR). Overall, 13 animals were considered positive by a direct detection method: seven out of 151 (4.6%) by MC-PCR and six out of 385 (1.6%) by bioassay, indicating the presence of viable parasites. As cattle that tested positive in the bioassay tested negative by MC-PCR and vice-versa, these results demonstrate a lack of concordance between the presence of viable parasites in liver and the detection of T. gondii DNA in diaphragm. In addition, the probability to detect T. gondii parasites or DNA in seropositive and seronegative cattle was comparable, demonstrating that serological testing by MAT or p30 immunoblot does not provide information about the presence of T. gondii parasites or DNA in cattle and therefore is not a reliable indicator of the risk for consumers.
Assuntos
Anticorpos Antiprotozoários/sangue , Doenças dos Bovinos/diagnóstico , Testes Diagnósticos de Rotina/métodos , Toxoplasma/isolamento & purificação , Toxoplasmose Animal/diagnóstico , Animais , Bovinos , Doenças dos Bovinos/parasitologia , Diafragma/parasitologia , Europa (Continente) , Imunoensaio/métodos , Imunoglobulina G/sangue , Fígado/parasitologia , Técnicas de Diagnóstico Molecular/métodos , Sensibilidade e Especificidade , Soro/imunologia , Soro/parasitologia , Toxoplasmose Animal/parasitologiaRESUMO
Toxoplasma gondii is a significant cause of abortion in sheep. Infection is picked up from the environment and if initiated during pregnancy may cause fetal mortality. Infected sheep remain persistently infected with tissue cysts in brain and muscle (meat), and are also immune and would not be expected to abort again. The live tachyzoite vaccine (Toxovax) protects against abortion and this allows the suggestion that it may also reduce or prevent tissue cyst development in muscle. If this were so it raises the question of whether the vaccine could be used to make meat safer for human consumption.
Assuntos
Doenças dos Ovinos/epidemiologia , Toxoplasmose Animal/epidemiologia , Aborto Animal/epidemiologia , Aborto Animal/etiologia , Aborto Animal/parasitologia , Aborto Animal/prevenção & controle , Ração Animal/parasitologia , Animais , Antiprotozoários/uso terapêutico , Gatos , Decoquinato/uso terapêutico , Feminino , Contaminação de Alimentos , Transmissão Vertical de Doenças Infecciosas , Parasitemia/epidemiologia , Parasitemia/parasitologia , Gravidez , Complicações Infecciosas na Gravidez/epidemiologia , Complicações Infecciosas na Gravidez/parasitologia , Complicações Infecciosas na Gravidez/veterinária , Vacinas Protozoárias , Ovinos , Doenças dos Ovinos/congênito , Doenças dos Ovinos/prevenção & controle , Doenças dos Ovinos/transmissão , Suínos/parasitologia , Toxoplasmose Animal/congênito , Toxoplasmose Animal/prevenção & controle , Toxoplasmose Animal/transmissão , Resultado do TratamentoRESUMO
A serial examination of three groups of cattle infected intravenously (iv) (Group 1, n=8) or subcutaneously (sc) (Group 2, n=8) with live Neospora caninum tachyzoites or with VERO cells (Group 3, n=8) at 70 days' gestation was carried out and the nature of the inflammatory responses in the placenta and the presence of parasite antigen were analysed. Immune cells expressing CD3, CD4, CD8, gamma delta (gammadelta) T-cell receptors (TCR), CD79alpha cytoplasmic (cy) (B cells) and NKp46 [natural killer (NK) cells] antigens were identified immunohistochemically and cells expressing mRNA for interferon-gamma (IFN-gamma) were labelled by in-situ hybridization. Intravenous inoculation caused mortality in all fetuses from 28 days post-inoculation (dpi) onwards. Subcutaneous inoculation caused mortality in 50% of the animals by 28dpi. Pathological changes in the placenta consisted of necrosis of fetal placental villi, necrosis and inflammation in adjacent areas of the maternal septum and inflammation at the base of the maternal caruncle. The inflammatory infiltrate consisted mainly of CD3(+) lymphocytes, dominated by CD4(+) and gammadelta TCR(+) cells, with CD8(+) cells present to a lesser extent. The results from the control group indicated fewer NK cells than those occurring in the placenta of human beings or mice. Infiltration of CD4(+) cells and NKp46(+) cells was observed in the caruncular base and septa 14 days after infection, whereas infiltration of gammadelta TCR(+) cells was observed from 28 dpi onwards. To our knowledge this is the first report on the presence and distribution of NK cells in the bovine placenta. Maternal inflammatory cells expressing mRNA for IFN-gamma were identified in animals inoculated with parasites iv or sc at 14 and 28 dpi, respectively. In the sc-inoculated dams with live fetuses at 28, 42 and 56dpi, there was no evidence of parasite antigen, infiltration of immune cells or production of IFN-gamma, suggesting that the parasite had not reached the placenta. The exact cause of fetal death was not established. Tissue destruction by the parasite may have occurred; in addition, there may have been a T helper 1 (Th-1) immune response to the neospora infection at the materno-fetal interface, resulting in infiltrations of CD4T cells, gammadelta T cells and NK cells and the subsequent production of IFN-gamma. It is possible that a pro-inflammatory Th-1 response early in gestation protects the dam by eliminating the parasite; however, it may lead to destruction of the placental tissues themselves and thus be incompatible with fetal survival.
Assuntos
Doenças dos Bovinos/parasitologia , Coccidiose/veterinária , Neospora/patogenicidade , Placenta/imunologia , Placenta/parasitologia , Prenhez/imunologia , Animais , Complexo CD3/genética , Complexo CD3/metabolismo , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD4-Positivos/patologia , Linfócitos T CD8-Positivos/metabolismo , Linfócitos T CD8-Positivos/patologia , Bovinos , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/metabolismo , Doenças dos Bovinos/patologia , Coccidiose/imunologia , Coccidiose/patologia , Feminino , Morte Fetal , Interferon gama/genética , Interferon gama/metabolismo , Neospora/imunologia , Placenta/metabolismo , Placenta/patologia , Gravidez , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Antígenos de Linfócitos T gama-delta/genética , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo , Receptores Imunológicos/genética , Receptores Imunológicos/metabolismo , Linfócitos T/imunologia , Linfócitos T/metabolismo , Linfócitos T/patologia , Células Th1/imunologia , Células Th1/metabolismo , Células Th1/patologiaRESUMO
Cryptosporidium transmission studies to date have concluded that adult cattle are not a significant source of oocysts contributing to clinical cryptosporidiosis in calves on farm. However current methods of sample processing have been optimised for calf faecal samples and may be less sensitive when used on adult samples due to lower numbers of oocysts and larger size of samples. A modified and novel method of oocyst extraction and concentration was developed and applied in an experiment involving spiking adult cattle faecal samples with known concentrations of Cryptosporidium oocysts. The results showed an increased sensitivity of detection from 100oocysts/g of faecal sample using conventional protocols to 5oocysts/g using the newly developed method. As it is important to be able to accurately assess the contribution of adult ruminants to the transmission of Cryptosporidium, both on farm and in the environment, the development of the techniques described here is likely to make an important contribution to Cryptosporidium transmission studies in future and in subsequent control strategies aimed at the reduction of Cryptosporidium infection in calves on farm.
Assuntos
Doenças dos Bovinos/parasitologia , Criptosporidiose/parasitologia , Cryptosporidium/isolamento & purificação , Fezes/parasitologia , Contagem de Ovos de Parasitas/veterinária , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Criptosporidiose/diagnóstico , Contagem de Ovos de Parasitas/métodos , Sensibilidade e EspecificidadeRESUMO
Avidity serological tests (avidity enzyme-linked immunosorbent assay [ELISA] and avidity Western blot) were developed and used to differentiate between acute (primary infection, reinfection, and recrudescence) and chronic Neospora caninum infection in cattle. In addition, the pattern of immunoglobulin G (IgG) avidity maturation against different specific antigens of N. caninum tachyzoites was studied. Sequential serum samples were collected from cattle naturally and experimentally infected with N. caninum. Four groups of experimentally infected cattle were included in the study and were representative of primary infection, reinfection, chronic infection, and noninfection. Serum samples were also collected from naturally infected cattle classified into nonaborting and aborting cows on the basis of clinical findings and serological profiles, and a third group composed of seronegative cows that seroconverted during the course of the experiment. All samples were tested by avidity ELISA and avidity Western blot. The IgG avidity ELISA allowed the discrimination between primary and chronic infection because all experimentally primary-infection cows showed low avidity indexes at week 4 postinfection (p.i.) compared with the high avidity values found at week 20 postinfection. However, this test did not allow the discrimination of reinfection or recrudescence from chronic infection. Regarding IgG avidity Western blot results, no antigenic markers correlating with acute (primary infection, recrudescence, and reinfection) or chronic infection were recognized. However, the 17-kD immunodominant antigen was mostly responsible for high avidity values obtained by avidity ELISA because it was intensively recognized by high-avidity antibodies in all chronically infected animals after urea treatment.
Assuntos
Western Blotting/veterinária , Doenças dos Bovinos/parasitologia , Coccidiose/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Neospora/imunologia , Aborto Animal/microbiologia , Aborto Animal/prevenção & controle , Doença Aguda , Animais , Anticorpos Antiprotozoários/sangue , Anticorpos Antiprotozoários/imunologia , Afinidade de Anticorpos , Western Blotting/métodos , Western Blotting/normas , Bovinos , Doenças dos Bovinos/diagnóstico , Doença Crônica , Coccidiose/diagnóstico , Coccidiose/parasitologia , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/normas , Epitopos/imunologia , Feminino , Imunoglobulina G/imunologia , GravidezRESUMO
The aim of the present study was to investigate and correlate the cell-mediated immune response and pathological changes at the maternal-fetal interface of Neospora-challenged pregnant cattle previously immunized with live and inactivated experimental vaccines. Pregnant heifers naïve to Neospora caninum were divided in 5 groups of 4 animals, each one immunized before mating: Group A heifers were intravenously (iv) immunized with 6.25 × 10(7) live tachyzoites of the NC-6 strain; group B heifers were immunized twice subcutaneously (sc) 3 weeks apart with native antigen extract of the NC-6 strain formulated with ISCOMs; group C heifers were sc immunized twice 3 weeks apart with three recombinant proteins (rNcSAG1, rNcHSP20, rNcGRA7) of the NC-1 strain formulated with ISCOMs; group D heifers were sc injected with sterile phosphate-buffered saline (PBS) and group E heifers received sc ISCOM-matrix (ISCOMs without antigen). All groups were iv-challenged with 4.7 × 10(7) NC-1 tachyzoites at 70 days of gestation. Heifers were culled at day 104 of gestation and placentomes were examined to evaluate lesions and local cellular immune responses using histopathology, immunohistochemistry and real time-PCR. Immunohistochemistry was performed using bovine leucocyte specific antibodies. Cytokine expression and levels (IFN-γ, IL-4, IL-10, IL-12 and TNF-α) were measured using real-time reverse transcription-PCR and ELISA, respectively. Minimal inflammation was observed in group A placentomes; while placentomes from group B, C, D and E had moderate to severe infiltration with CD3(+), CD4(+), γδ-T cells, CD8(+) cells and macrophages being more numerous in groups B and E placentomes, when compared with groups C and D (P<0.001). Cytokine levels were significantly increased in the caruncles of animals of groups B and C in comparison with the other animal groups (P < 0.001). The results from this study showed that the strongest cellular immune responses were observed in the placentomes of animals that were immunized with inactivated vaccines (groups B and C) and in the placentomes of animals that were sc-sham-inoculated (groups D and E). On the other hand, animals that were immunized with live tachyzoites showed a milder immune cell infiltration to the placenta possibly due to the existence of a protective systemic maternal immune response that helped to minimize N. caninum infection at the maternal-fetal interface.
Assuntos
Doenças dos Bovinos/imunologia , Coccidiose/veterinária , Imunidade Celular/imunologia , Neospora/imunologia , Placenta/imunologia , Vacinas Protozoárias/imunologia , Animais , Bovinos , Doenças dos Bovinos/prevenção & controle , Coccidiose/imunologia , Citocinas/sangue , Feminino , Gravidez , Vacinas Protozoárias/normas , Vacinação/veterinária , Vacinas de Produtos Inativados/imunologia , Vacinas de Produtos Inativados/normasRESUMO
The presence of Toxoplasma gondii in blood, brain, cardiac muscle and skeletal muscle (gracillis and psoas) of sheep 6 weeks after experimental infection with 10(5), 10(4) and 10(3) T. gondii oocysts was determined using the PCR technique. The study demonstrates that oral infection of sheep with T. gondii oocysts of the M3 isolate results in parasites being detectable in tissues 6 weeks p.i. The PCR detection was much more sensitive than histological detection. Parasite DNA was detected more frequently and consistently in the group of sheep given 10(5) oocysts compared with those given 10(3) oocysts. The brain and heart were most frequently infected compared with the other tissues.
Assuntos
Doenças dos Ovinos/parasitologia , Ovinos/parasitologia , Toxoplasma/isolamento & purificação , Toxoplasmose Animal/parasitologia , Fatores Etários , Animais , Anticorpos Antiprotozoários/sangue , Especificidade de Anticorpos , Gatos , DNA de Protozoário/análise , Ensaio de Imunoadsorção Enzimática , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Imunoglobulina G/sangue , Masculino , Camundongos , Especificidade de Órgãos , Parasitemia/imunologia , Parasitemia/veterinária , Reação em Cadeia da Polimerase , Doenças dos Ovinos/imunologia , Organismos Livres de Patógenos Específicos , Fatores de Tempo , Toxoplasma/imunologia , Toxoplasmose Animal/imunologiaRESUMO
In spite of the global importance of neosporosis as a cause of bovine abortion, there is very little information about its economic consequences. The economic costs are a product of estimations of the quantity of the effects attributable to Neospora infection, and the particular unit costs of those effects. In this brief review, which arose from a workshop on the economics of coccidiosis held at the COST 820 meeting, Toledo 1998, we discuss the possible effects of neosporosis which are of economic significance and summarise the available estimates of their magnitude to provide a basis for further economic analysis. Neospora infection has been associated with abortion, increased culling and reduced milk yield. In addition, it has been diagnosed in cases of stillbirth and neonatal mortality, it is likely to contribute to early foetal death and resorption and it is responsible for a reduction in the value of female breeding cattle. In quantifying the role of Neospora, it is important that epidemiologically based, case-controlled studies are conducted because, given the extreme efficiency with which bovine Neospora infection is vertically transmitted, demonstration of prevalence of infection in affected animals (including foetuses) is not a true indicator of the significance of this disease. Relatively few epidemiological studies have been conducted, but in investigations in the USA, Holland and Britain, infected cows have been shown to be about three times more likely to abort than non-infected cattle. In the UK this approach has been used to estimate the proportion of abortions in the national dairy population which may be attributable to Neospora caninum.
Assuntos
Aborto Animal/economia , Criação de Animais Domésticos/economia , Doenças dos Bovinos/economia , Coccidiose/veterinária , Neospora , Complicações Parasitárias na Gravidez/veterinária , Aborto Animal/epidemiologia , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Coccidiose/economia , Coccidiose/epidemiologia , Efeitos Psicossociais da Doença , Feminino , Gravidez , Complicações Parasitárias na Gravidez/economia , Complicações Parasitárias na Gravidez/epidemiologiaRESUMO
In this study we were interested to determine whether infection of cattle prior to pregnancy would afford any protection to the foetus if the dams were challenged with Neospora caninum at mid-gestation. The experiment comprised four groups of cattle: group 1, uninfected controls; group 2, inoculated with N. caninum tachyzoites 6 weeks prior to mating and then challenged with N. caninum at mid-gestation; group 3, naive cattle challenged with N. caninum at mid-gestation and group 4 were infected with N. caninum prior to mating and left unchallenged throughout pregnancy. Positive cell-mediated and humoral immune responses to N. caninum were recorded in groups 2 and 4 prior to pregnancy and in groups 2, 3 and 4 following challenge at mid-gestation. However there was a marked down regulation of the cell-mediated immune response in all groups around mid-gestation. There was a significant increase in rectal temperature response in animals in group 3 compared to group 2 following challenge but no other clinical symptoms of disease were recorded and all cattle proceeded to calving. At calving, pre-colostral blood samples were negative for antibodies to N. caninum in all the calves born to dams in groups 1, 2 and 4. In contrast, all the calves born to dams in group 3 had high levels of specific antibody to N. caninum indicating that they had been exposed to the parasite in utero. At post-mortem N. caninum DNA was detected in CNS, thymus and placental cotyledon samples in calves from group 3. All tissue samples from calves in the other 3 groups were negative for N. caninum DNA with the exception of one calf from group 2 where specific DNA was detected in a sample of spinal cord. These results suggest that the immune response generated in the dams in group 2 prior to pregnancy had protected against vertical transmission of the parasite following challenge at mid-gestation.
Assuntos
Doenças dos Bovinos/transmissão , Coccidiose/veterinária , Transmissão Vertical de Doenças Infecciosas/veterinária , Neospora/crescimento & desenvolvimento , Animais , Animais Recém-Nascidos , Anticorpos Antiprotozoários/sangue , Temperatura Corporal , Bovinos , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/parasitologia , Coccidiose/imunologia , Coccidiose/transmissão , DNA de Protozoário/química , Feminino , Histocitoquímica/veterinária , Transmissão Vertical de Doenças Infecciosas/prevenção & controle , Interferon gama/imunologia , Masculino , Leite/imunologia , Neospora/genética , Neospora/imunologia , Placenta/parasitologia , Placenta/patologia , Reação em Cadeia da Polimerase/veterinária , GravidezRESUMO
In recent years, neosporosis has been identified as a major cause of abortion in dairy and beef cattle. Although the disease has been described worldwide, there is a Jack of information concerning the prevalence of this infection in different cattle production systems. The aim of this study was to investigate the seroprevalence of Neospora caninum infection in a representative area of beef and dairy cattle production in Spain. A cross-sectional study was undertaken in which herds constituted the initial sampling unit and two strata (dairy and beef herds) were considered. Using a 95% level of confidence and setting 5% (beef) and 5.4% (dairy) error limits, 216 beef and 143 dairy herds were randomly selected and sampled. Nine animals (> 1 year old) were randomly sampled in each herd to detect the presence of the infection. A herd was considered infected when at least one animal was seropositive. In total, serum samples from 1121 dairy and 1712 beef animals were collected and tested for specific anti-N. caninum IgG using an ELISA. Specific antibodies were detected in 55.1% (119/216) beef and 83.2% (119/143) dairy herds. Individual prevalences obtained were 17.9% (306/1712) for beef and 35.9% (402/1121) for dairy animals. Presence of N. caninum infection was higher in dairy than in beef herds and the association between infection and the cattle production system (dairy or beef) was statistically significant [(chi2)Y= 29.21, P < 0.001, OR = 4.04 (2.35-6.99)]. Herd size of dairy cattle did not appear to be associated with N. caninum infection. On the contrary, infection was associated with herd size in beef cattle (chi2 = 12.79, P < 0.01). Finally, no association was found between replacement or pasture management and infection in beef herds.
Assuntos
Anticorpos Antiprotozoários/sangue , Doenças dos Bovinos/epidemiologia , Coccidiose/veterinária , Neospora/imunologia , Criação de Animais Domésticos/métodos , Animais , Bovinos , Coccidiose/epidemiologia , Estudos Transversais , Indústria de Laticínios , Ensaio de Imunoadsorção Enzimática , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Masculino , Neospora/isolamento & purificação , Estudos Soroepidemiológicos , Espanha/epidemiologiaRESUMO
Neospora caninum is a recently recognized protozoan parasite which has been described as causing a neuromuscular paralysis in dogs and is emerging as a major cause of bovine infertility and abortion worldwide. The parasite is known to infect a range of warm blooded animals but the disease predominates in dogs and cattle. It is not yet known if N. caninum can infect and cause disease in people. The dog has recently been identified as the definitive host and the parasite may be transmitted through the ingestion of oocysts or congenitally from mother to fetus. N. caninum is known to infect red foxes (Vulpes vulpes) and coyotes (Canis latrans) and the role of wildlife species as reservoirs of infection requires further investigation. Little is known about the range of parasite genotypes within the environment or the variation in virulence between different strains. RAPD-PCR analysis of geographically distinct bovine and canine isolates has revealed little genetic variation. Epidemiological studies from different areas of the world have investigated the importance of N. caninum as an abortifacient agent and longitudinal studies have shown the high rate (approximately 80%) of congenital transmission within infected herds. Information on the rates of repeat abortion due to neosporosis are less well defined however current estimates put this at 5% suggesting that cattle may develop some form of protective immunity against N. caninum-induced abortion. Diagnosis of the disease is based upon detection of the parasite in the tissues, most commonly using immunohistochemistry with additional information provided by serology. However, although positive fetal serology is a strong indicator of exposure to the parasite, care should be taken in the interpretation of maternal serology. As we understand more about the epidemiology of neosporosis we are also better able to interpret the results of diagnostic tests. The mere presence of the parasite does not necessarily infer that this was the primary cause of abortion. CD4+ T-cells, interferon gamma and macrophages have all been found to significantly inhibit multiplication of N. caninum tachyzoites. The nature of a protective immune response and its modulation in the pregnant animal is discussed.
Assuntos
Doenças dos Bovinos/epidemiologia , Coccidiose/veterinária , Doenças do Cão/epidemiologia , Neospora , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/imunologia , Coccidiose/epidemiologia , Coccidiose/imunologia , Doenças do Cão/diagnóstico , Doenças do Cão/imunologia , Cães , Técnica de Amplificação ao Acaso de DNA PolimórficoRESUMO
An open study was carried out in 40 elderly patients suffering from osteoarthrosis of the hip to assess the effectiveness of 200 mg flurbiprofen daily, fiven either in tablet or liquid suspension form, over a period of 4 weeks. Statistical evaluation of the results in the 19 patients admitted to analysis showed that flurbiprofen produced significant improvement in daytime and overnight pain, and did not impair their mental status, as assessed on a modified Goldfarb scale. No significant differences were found between the response of the two groups, except for severity of night pain. Fourteen of the 19 patients included in the analysis reported side-effects, as did 18 of the 21 patients who were excluded. Only 7 patients (4 on tablets, 3 on liquid), however, were withdrawn from the trial for this reason.
Assuntos
Flurbiprofeno/administração & dosagem , Osteoartrite/tratamento farmacológico , Propionatos/administração & dosagem , Idoso , Envelhecimento , Formas de Dosagem , Feminino , Flurbiprofeno/efeitos adversos , Flurbiprofeno/uso terapêutico , Articulação do Quadril , Humanos , Masculino , Osteoartrite/psicologia , Cuidados Paliativos , Cooperação do PacienteRESUMO
The protozoan parasite Toxoplasma gondii is capable of infecting all warm blooded animals; however, the consequences of infection are very variable between different species of animal. Marsupials and New World monkeys, which have evolved largely separately from the cat, the definitive host of the parasite, are among the most vulnerable species where infection with T. gondii can prove fatal. In more resistant species such as humans and sheep, infection is generally unapparent, provoking only mild symptoms; thereafter the host remains infected for life. However, when the immune system is compromised, such as in the immunologically immature fetus, infection with the parasite can have very serious consequences. Much of the work examining host immune responses has been done using experimentally infected mice. While there are many advantages in using this experimental model, care should be taken in extrapolating results from mice to other species. Mice are extremely vulnerable to the consequences of infection with T. gondii., and their use to further our understanding of congenital toxoplasmosis may not be ideal, as fetal infection can occur in successive pregnancies. This is not the case in rats or sheep; they are more resistant to the disease and therefore may provide a more relevant model for human congenital toxoplasmosis. Studies of host immune responses have emphasised the importance of the cytokine interferon gamma (IFN gamma) in resistance to T. gondii. The efficiency of induction of this cytokine may be critical for determining the outcome of the host-parasite relationship.