RESUMO
Susceptability of Ascaridia galli to benzimidazole (BZ) was investigated using faecal egg count reduction test (FECRT), in ovo larval development test (LDT) and genetic markers (mutations at codons 167, 198 and 200 of ß-tubulin gene). Six flocks (F1-F6) of a commercial laying hen farm with different number of exposure to BZ were recruited. The FECR was calculated by analyzing individual faeces (F1, F2, F4 and F5) before and 10 days after treatment. The LDT was performed on parasite eggs from pooled samples from F1 to F6 and LC50 and LC99 were calculated. DNA was extracted from 120 worms and sequenced for ß-tubulin gene. In all flocks, the FECRs were above 95% (lower CI above 90%). No significant difference was observed (p > 0·05) among obtained LC50 (F1/F4 and F2/F5 vs F3/F6) in the LDT. However, LC50 and LC99 were higher than suggested values for declaration of resistance in other nematode species. No variation was observed in codon positions involved in BZ resistance. Overall, our results indicated lack of evidence of resistance to BZ in A. galli. More research is needed to confirm these results and to further optimize the existing tools for detection and monitoring of anthelmintic resistance in A. galli.
Assuntos
Antinematódeos/farmacologia , Ascaridia/efeitos dos fármacos , Ascaridíase/veterinária , Benzimidazóis/farmacologia , Galinhas , Doenças das Aves Domésticas/tratamento farmacológico , Animais , Ascaridia/genética , Ascaridia/crescimento & desenvolvimento , Ascaridíase/tratamento farmacológico , Códon/efeitos dos fármacos , Códon/genética , Códon/metabolismo , Fezes/parasitologia , Feminino , Marcadores Genéticos/genética , Proteínas de Helminto/genética , Proteínas de Helminto/metabolismo , Larva/efeitos dos fármacos , Larva/genética , Larva/crescimento & desenvolvimento , Masculino , Contagem de Ovos de Parasitas/veterinária , Análise de Sequência de DNA/veterinária , Tubulina (Proteína)/genética , Tubulina (Proteína)/metabolismoRESUMO
Haematophagous mites were collected from the vent region and plumage of chickens in six hobby flocks of ornamental breeds in Sweden, one of which included turkeys. Soiled vent skin and feathers, dermatitis, hyperkeratosis, skin necroses and ulcers were observed in 12 necropsied birds from two of the flocks. The mites were identified as the northern fowl mite Ornithonyssus sylviarum (Mesostigmata: Macronyssidae). This was supported by sequence analysis of a 642-bp region in the mitochondrial cytochrome oxidase subunit 1 (COI) gene (COI) in mites collected from five flocks, which showed 97-99% sequence similarity to O. sylviarum by blast analysis. Pairwise sequence comparisons revealed nucleotide variations in the range of 0-2.8%, whereas amino acid sequences were highly conserved. This paper represents one of very few records of O. sylviarum in European poultry, and is the first to report COI sequence data for O. sylviarum from poultry in Europe.
Assuntos
Galinhas , Infestações por Ácaros/veterinária , Ácaros/classificação , Doenças das Aves Domésticas/parasitologia , Animais , Sequência de Bases , DNA/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Infestações por Ácaros/epidemiologia , Infestações por Ácaros/parasitologia , Ácaros/enzimologia , Ácaros/genética , Filogenia , Doenças das Aves Domésticas/epidemiologia , Suécia/epidemiologiaRESUMO
1. Vancomycin-resistant Enterococcus faecium (VRE(fm)) has recently spread among Swedish broiler farms. The objectives were to investigate VRE(fm) persistence within barns between flocks, and to determine whether day-old chicks, feed or forklift trucks used for loading crates could be identified as a means of transmission. 2. Faeces were collected for selective culture from 12 farms (9 culture-positive, 3 culture-negative as determined by prior monitoring), and samples were collected from the barn environment before and after cleaning and disinfection, from forklift tyres, hatcheries and feed. 3. VRE(fm) was isolated only from previously known VRE(fm)-positive farms. The proportions of culture-positive environmental samples were 75% (9 out of 9 farms) prior to and 31% (7 out of 9 farms) after cleaning/disinfection. Five out of 6 samples from forklift tyres were culture-positive. No VRE(fm) were isolated from feed or hatcheries. The majority of 27 vanA gene positive isolates showed similar banding patterns by SmaI restriction digestion and pulse-field gel electrophoresis. No consistent differences were observed regarding management between VRE(fm)-positive and negative farms. 4. We conclude that VRE(fm) contaminates barns and remains present between flocks. Forklift trucks are one possible means of transmission between farms.
Assuntos
Galinhas , Enterococcus faecium/genética , Infecções por Bactérias Gram-Positivas/veterinária , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/transmissão , Resistência a Vancomicina , Ração Animal/microbiologia , Criação de Animais Domésticos , Animais , Proteínas de Bactérias/genética , Carbono-Oxigênio Ligases/genética , Contagem de Colônia Microbiana/veterinária , Desoxirribonucleases de Sítio Específico do Tipo II/metabolismo , Eletroforese em Gel de Campo Pulsado/veterinária , Enterococcus faecium/efeitos dos fármacos , Enterococcus faecium/isolamento & purificação , Fezes/microbiologia , Infecções por Bactérias Gram-Positivas/epidemiologia , Infecções por Bactérias Gram-Positivas/microbiologia , Infecções por Bactérias Gram-Positivas/transmissão , Testes de Sensibilidade Microbiana/veterinária , Doenças das Aves Domésticas/microbiologia , Prevalência , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Suécia/epidemiologia , Meios de TransporteRESUMO
Since the EU ban on battery cages, many studies have listed Ascaridia galli and Heterakis gallinarum as the most common roundworms in the European laying hen population. A complicating factor is that the eggs of these parasites are almost identical. Thus, lack of molecular diagnostic approaches has driven epidemiological studies to take on necropsy for species discrimination, which is labor and cost intensive. Here, we describe a novel diagnostic tool based on droplet digital PCR for simultaneous identification and absolute quantification of the eggs of both of these ascarids in chickens' droppings using two different genus-specific primer-probe sets targeting the second internal transcribed spacer region (ITS-2) in the nuclear ribosomal (rRNA) gene array. No cross-reaction was observed when different combinations of DNA and species-specific primers and probes were tested. The lowest obtained frequency threshold for the detection of H. gallinarum in the presence of a constant A. galli DNA concentration was determined to be 0.8 %. After validation, we used the assay to analyze field samples collected from several Swedish laying hen farms. Out of 134 samples, 86 (64 %) were positive for A. galli while 11 (8.3 %) samples were positive for H. gallinarum. These samples were initially analyzed with flotation technique for detection of ascarid eggs. The results of the Cohen's kappa indicated substantial agreement (85.8 %) between the two tests. In conclusion, we have validated a novel molecular-based diagnostic tool for quantification and differentiation between intestinal parasites of major importance in chickens with high precision. Although this study focuses on identification of parasites of laying hens, the findings may well have a bearing on all types of chicken production systems. The present study lays the groundwork for future research into epidemiology of these two important chicken parasite species.
Assuntos
Ascaridia , Ascaridíase , Nematoides , Infecções por Nematoides , Reação em Cadeia da Polimerase , Doenças das Aves Domésticas , Animais , Ascaridia/genética , Ascaridíase/diagnóstico , Ascaridíase/parasitologia , Ascaridíase/veterinária , Galinhas , Diagnóstico Diferencial , Fezes/parasitologia , Feminino , Nematoides/genética , Infecções por Nematoides/diagnóstico , Infecções por Nematoides/parasitologia , Infecções por Nematoides/veterinária , Óvulo , Reação em Cadeia da Polimerase/normas , Reação em Cadeia da Polimerase/veterinária , Doenças das Aves Domésticas/diagnóstico , Doenças das Aves Domésticas/parasitologiaRESUMO
Worm control is an important aspect of the successful management of the egg production industry. Of particular concern is Ascaridia galli, which at high parasite loads affect health and production in layers. Application of a targeted treatment strategy (TT) to control A. galli has shown promise. The purpose of this study was to explore the effect of such a strategy on welfare indicators and production performance of layers. Six flocks (F1-6) on a commercial farm were allocated to three treatment groups. Flocks F1 and F4 were treated (TT) with fenbendazole at 22, 27 and 36 weeks post-placement (WPP). Flocks F2 and F5 were treated at 27 WPP (conventional treatment, CT) and hens in flocks F3 and F6 served as untreated (UT) control groups. At 19, 35 and 45 WPP twenty-five hens plus thirty eggs per flock were randomly selected. Hens were weighed and their plumage conditions (PC) were assessed. The eggs were subjected to various external and interior quality analyses. Production data such as number of eggs/hen/week, egg mass and feed conversion ratio (FCR) were calculated from raw data obtained from all flocks on a weekly basis. The number of eggs/hen/week, egg mass and FCR were higher (Pâ¯<â¯0.05) in the TT flocks and hens had better PC both at 35 and 45 WPP compared with other flocks. No differences in body weight and physical egg quality were observed between groups except for egg shell strength which was higher (Pâ¯<â¯0.05) in the CT flocks. These data suggest that better production performance and plumage, which suggests improved health, can be achieved through the application of a TT strategy. The insights gained from this research should help to justify the extra cost and labor associated with the TT strategy.
Assuntos
Ascaridíase/veterinária , Galinhas , Doenças das Aves Domésticas/prevenção & controle , Criação de Animais Domésticos , Animais , Ascaridia/fisiologia , Ascaridíase/parasitologia , Ascaridíase/prevenção & controle , Galinhas/fisiologia , Feminino , Óvulo/fisiologia , Doenças das Aves Domésticas/parasitologia , ReproduçãoRESUMO
Several species of intestinal spirochaetes, Brachyspira (B.) alvinipulli, B. intermedia and B. pilosicoli, may cause reduced egg production and faecal staining of eggshells in chickens. The aim of this study was to characterize potentially pathogenic and presumably non-pathogenic Brachyspira spp. from commercial laying hens. Selective culture, phenotyping, PCR and 16S rRNA gene sequencing were used and clinical data were collected. Phenotypic profiles were obtained for 489 isolates and 351 isolates obtained after subculture, and 30 isolates were selected for molecular characterization. Seven isolates were positive by a B. intermedia-specific PCR based on the nox gene, and two were positive in a B. hyodysenteriae-specific 23S rRNA gene based PCR. By comparative phylogenetic analysis in combination with PCR and phenotyping, seven isolates were identified as B. intermedia, eight isolates as B. innocens, five as B. murdochii, and three isolates each as B. alvinipulli and "B. pulli". The remaining four isolates could not be assigned to any presently recognized species. Co-infection with several species or genetic variants of Brachyspira spp. were detected in some flocks and samples, suggesting a high level of diversity. Organic flocks with access to outdoor areas were at higher risk (RR=2.3; 95% CI 1.5-3.6) for being colonized than chickens in other housing systems. No significant differences between colonized and non-colonized flocks were found regarding clinical parameters, i.e. mortality, egg production, faecally contaminated eggshells, and wet litter. Our results show that a combination of traditional laboratory diagnostics, molecular tests and phylogeny is needed for identification of Brachyspira sp. from chickens.
Assuntos
Brachyspira/classificação , Brachyspira/genética , Galinhas , Infecções por Bactérias Gram-Negativas/veterinária , Abrigo para Animais , Doenças das Aves Domésticas/microbiologia , Animais , Feminino , Infecções por Bactérias Gram-Negativas/epidemiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Oviposição , Filogenia , Reação em Cadeia da Polimerase , Doenças das Aves Domésticas/epidemiologia , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Especificidade da Espécie , Suécia/epidemiologiaRESUMO
The reliability of the results of in vitro studies such as detection of anthelmintic resistance often depends on the ability of the parasite eggs to develop under laboratory conditions. The aim of this study was to assess the embryonation capability of the chicken roundworm Ascaridia galli eggs after storage under different conditions. Two storage media for parasite eggs were used; faeces or water. Eggs in petri dishes (90 dishes in total) containing faces or water media were first exposed either to aerobic or anaerobic conditions at different temperatures (4⯰C /+O2, 4⯰C /-O2, 25⯰C/-O2) for a maximum of 72 days. Every second week, materials from petri dishes in triplicates were recovered and incubated aerobically for two weeks at 25⯰C. After the incubation, 200-300 eggs from each petri shish (sampling unit) were counted and the number of embryonated eggs was determined. Data was analyzed in R (version 3.4.3) A logistic regression model with the probability of an egg to embryonate as dependent variable and conditions, storage medium and time points as fixed effects with quasibinomial distribution was run. Least-square means were calculated and pairwise comparisons were made with the fixed effect factors (condition, storage medium and time point). Eggs in faeces had a significantly (pâ¯Ëâ¯0.05) higher embryonation than those in water, irrespective of storage conditions. At 4⯰C embryonation tended to decline over time under aerobic conditions irrespective of the storage medium, whereas it remained constant following storage at anaerobic conditions. In contrast, anaerobic storage at the 25⯰C negatively affected egg development in both media, except for day 14 in faeces. Our major finding was that eggs in faeces under anaerobic conditions and at 4⯰C retained the highest rate of development, with a minimum decline in their developmental capacity over time compared to cleaned eggs stored in water.
Assuntos
Ascaridia/crescimento & desenvolvimento , Manejo de Espécimes/métodos , Anaerobiose , Animais , Galinhas/parasitologia , Fezes/parasitologia , Óvulo/crescimento & desenvolvimentoRESUMO
The efficacy of a sustainable deworming strategy based on targeted treatments (TT) against Ascaridia galli was investigated for the first time in laying hen flocks on a Swedish commercial farm. Three experimental protocols with different levels of treatment, e.g. targeted treatment (TT), conventional treatment (CT) and untreated (UT), were tested in randomly allocated flocks of two different bird hybrids. Every second week faecal egg counts (FECs) were determined from pooled faecal materials collected on trays (20×27cm) placed for a maximum of 12h on the litter belts. In the TT, anthelmintic administration (fenbendazole, 1mg/kg body weight for 5days) started at 22 weeks post placement (wpp) and was repeated twice when the pooled FECs surpassed the assigned threshold of 200 egg per gram faeces (EPG). The CT flocks were treated once at 27wpp using the same anthelmintic. Hens in the UT were not dewormed and served as controls. Additionally, FECs on cloacal contents, worm fecundity and worm burdens were determined at 19, 35 and 45wpp. None of the flocks became infected until after 16wpp. The cumulative pooled FECs at the end of the study were significantly (p<0.01) lower in the TT compared to both CT and UT. Although repeated treatment in the TT protocol did not affect the fecundity, a worm density-dependent increase in fecundity was observed. Cloacal FECs and the number of adult A. galli in TT at 35 and 45wpp were significantly lower compared to other flocks. This study provides evidence that the TT strategy is better in terms of lower worm burden and decreased cumulative environmental parasite egg numbers compared to CT strategy. The TT strategy should be considered as an alternative to the CT strategy with regard to A. galli control in commercial laying hens.
Assuntos
Anti-Helmínticos/uso terapêutico , Ascaridia/efeitos dos fármacos , Ascaridíase/veterinária , Galinhas/parasitologia , Doenças das Aves Domésticas/tratamento farmacológico , Animais , Anti-Helmínticos/administração & dosagem , Anti-Helmínticos/farmacologia , Ascaridia/fisiologia , Ascaridíase/tratamento farmacológico , Ascaridíase/parasitologia , Cloaca/parasitologia , Fezes/parasitologia , Feminino , Fertilidade/efeitos dos fármacos , Intestinos/parasitologia , Masculino , Contagem de Ovos de Parasitas/veterinária , Doenças das Aves Domésticas/parasitologiaRESUMO
Infection with the poultry roundworm Ascaridia galli has increased in European countries due to the ban on battery cages. This study was conducted in two commercial laying hen flocks (F1 & F2) on different farms in central Sweden. The aims were to (1) investigate the efficacy of flubendazole (FLBZ, 1.43 mg/kg administered in drinking water for 7 days) against adult and larval stages including histotrophic larvae of A. galli, and (2) determine how long it took before the flocks were reinfected after deworming. Accordingly, 180 randomly selected hens were sacrificed before drug administration (bd), on day 3 and 7 during drug administration (dd), and on a weekly basis for up to five weeks post drug administration (pd). Intestinal contents and cloacal materials of each hen plus pooled faecal samples from manure belts were investigated to assess the worm burden and the parasite egg per gram faeces (epg). Additionally, drinking water, and serum and gastrointestinal digesta content samples obtained from ten treated animals were analyzed by HPLC to measure FLBZ and its reduced (R-FLBZ) and hydrolyzed (H-FLBZ) metabolites. No parasite eggs were observed in cloacal samples on day 21 and 28 pd on F1 and on day 21 pd on F2. The epg in manure decreased by 65% and 88% on day 3 dd and by 99% and 97% on day 35 pd on F1 and F2 respectively. Mean FLBZ concentrations quantified in duodenal contents ranged between 0.50 and 0.79 µg/g. Although, no histotrophic larvae were found dd, they reappeared one week pd (7 ± 7 F1, 0.5 ± 0.5 F2). Adult worms were found in both flocks before drug administration (44 ± 20 F1, 35 ± 25 F2), on day 3 dd (4 ± 3 F1, 2 ± 2 F2), and then not until day 35 (0.2 ± 0.6) on F1 and day 28 (0.4 ± 0.9) pd on F2. Thus, the only period in which no A. galli were found was on day 7 dd. Although FLBZ was highly efficient our results indicate that the birds were reinfected already within one week pd.
Assuntos
Ascaridíase/veterinária , Mebendazol/análogos & derivados , Doenças das Aves Domésticas/tratamento farmacológico , Animais , Ascaridia/fisiologia , Ascaridíase/tratamento farmacológico , Galinhas , Fezes/parasitologia , Feminino , Conteúdo Gastrointestinal/química , Larva , Mebendazol/uso terapêutico , Carga Parasitária , Suécia , Resultado do TratamentoRESUMO
We studied by immunocytochemistry 573 tissue and 106 cytologic samples of human tumors, non-neoplastic proliferative lesions and normal tissues with the monoclonal antibody (Mab) A-80 that recognizes a mucinous glycoprotein from the colon carcinoma cell line LS-174T. The spectrum of benign and malignant breast lesions was studied as were epithelial tumors of the colon, stomach, pancreas, lung, salivary glands, thyroid, prostate, kidney, endometrium, skin and mesothelium; non-epithelial tumors included lymphomas, melanomas, gliomas, meningiomas, and sarcomas of soft tissue and bone. With a single exception, breast carcinomas regardless of histologic type were reactive while few fibroadenomas stained weakly and focally. In fibrocystic disease, the presence and intensity of the reactivity paralleled the severity of the epithelial proliferation, e.g. staining was strong in foci of severe or atypical hyperplasia, borderline lesions and carcinomas in situ; apocrine metaplasia stained often but less strongly. Barrett's mucosa, colonic polyps and most gastric and colonic carcinomas stained regardless of glandular features while small cell neuroendocrine carcinomas did not. Adenocarcinomas of the pancreas and lung, and a subset of large cell lung carcinomas reacted whereas neuroendocrine carcinomas of those sites did not. Carcinomas of endometrium, ovary and prostate reacted variably whereas thyroid and renal carcinomas and mesotheliomas were either negative or weakly reactive despite the presence of glands. Lymphomas, skin adnexal tumors, nevi, schwannomas, melanomas, gliomas and sarcomas generally did not react but occasional A-80-positive cells were seen in rare sarcomas and meningiomas. Immunostaining patterns in cytologic specimens were similar to the aforementioned. We conclude that Mab A-80 is an excellent marker for breast carcinomas, and for certain proliferative forms of fibrocystic disease that may precede or be associated with carcinomatous transformation. In colonic, pulmonary and gastric carcinomas, staining with Mab A-80 revealed exocrine features regardless of the absence of glands whereas in renal and thyroid carcinomas and in mesotheliomas staining was focal and weak or absent irrespective of glandular features. We suggest that Mab A-80 is a very promising immunolabel for select exocrine carcinomas, and for some of the dysplasias that may precede their development; its ease of application on tissue sections and cytologic specimens should broaden its usefulness.
Assuntos
Anticorpos Monoclonais , Mucinas/metabolismo , Neoplasias/metabolismo , Anticorpos Monoclonais/imunologia , Biomarcadores Tumorais/análise , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Carcinoma/metabolismo , Carcinoma/patologia , Epitélio/metabolismo , Epitélio/patologia , Doença da Mama Fibrocística/metabolismo , Doença da Mama Fibrocística/patologia , Neoplasias Gastrointestinais/metabolismo , Neoplasias Gastrointestinais/patologia , Humanos , Imuno-Histoquímica , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Mucinas/imunologia , Neoplasias/patologiaRESUMO
BACKGROUND: The purpose of this study was to determine by immunocytochemistry the relative incidence and clinicopathologic characteristics of neuroendocrine carcinomas of the stomach. METHODS: Sections from paraffin blocks from 81 patients who had undergone resection of carcinomas of the stomach were immunostained with a battery of neuroendocrine differentiation markers and with A-80, a marker of exocrine differentiation. The clinical and pathologic data of the 12 patients diagnosed with neuroendocrine carcinomas of the stomach were analyzed. RESULTS: The 10 men and two women ranged from 53 to 81 years of age (median, 69 years). Procedures performed included distal subtotal gastrectomy in eight patients and total gastrectomy in four patients. Pathologic stages were stage I, one patient; stage III, four patients; and stage IV, seven patients. Metastatic sites included regional nodes, 11 patients; liver, four patients; and bone, one patient. Adjunct treatment included multiagent chemotherapy plus radiotherapy, four patients; and only radiotherapy, one patient. Eleven patients died of disease 1 to 27 months after diagnosis with an overall median survival of 15 months. Three groups of neuroendocrine carcinomas were identified based on immunostaining patterns. These included pure neuroendocrine carcinomas, two patients; neuroendocrine carcinomas with occasional exocrine cells, three patients; and mixed neuroendocrine-exocrine carcinomas, seven patients. CONCLUSIONS: (1) The relative incidence of neuroendocrine differentiation in carcinomas of the stomach is higher than is generally recognized. (2) Neuroendocrine gastric carcinomas behave aggressively and display numerous structural and functional similarities with their colonic, extrahepatic biliary tract, and pulmonary counterparts.
Assuntos
Neoplasias das Glândulas Endócrinas/patologia , Neoplasias do Sistema Nervoso/patologia , Neoplasias Gástricas/patologia , Idoso , Neoplasias das Glândulas Endócrinas/metabolismo , Neoplasias das Glândulas Endócrinas/cirurgia , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Neoplasias do Sistema Nervoso/metabolismo , Neoplasias do Sistema Nervoso/cirurgia , Neoplasias Gástricas/metabolismoRESUMO
Only limited data concerning the prevalence of intestinal spirochetes are available in game birds. This paper describes the prevalence and biochemical reactions of spirochetes isolated from 25 common partridges, 7 pheasants and 16 mallards originating from nine Swedish game-bird farms. The birds, which had been submitted for post-mortem examination due to various problems, showed a variety of underlying diseases. Additionally, fecal droppings from 22 common partridges, 20 pheasants and 20 mallards obtained at one of the farms were included in the study. Intestinal spirochetes were isolated from 85.4% of the game birds and from 71% of the fecal droppings. Seven biochemical types were identified. Seventeen per cent of all isolates were classified as Brachyspira pilosicoli and 3% as B. intermedia. One isolate showed strong beta-hemolysis and a positive indole reaction that is consistent with B. hyodysenteriae. In addition, three previously unknown biochemical types were found. Unclassified spirochetes in presumed mixed cultures were observed in 21% of all samples of fecal droppings. Histologic examination of spirochete-positive birds revealed numerous spirochetes in the lumen and crypts of the cecum, in some cases densely adhered by one end to the luminal surface. The significance of the findings is discussed.
Assuntos
Doenças das Aves/epidemiologia , Brachyspira/isolamento & purificação , Infecções por Spirochaetales/veterinária , Animais , Animais Selvagens , Aderência Bacteriana , Doenças das Aves/microbiologia , Aves , Brachyspira/classificação , Ceco/microbiologia , Fezes/microbiologia , Prevalência , Infecções por Spirochaetales/epidemiologia , Suécia/epidemiologiaRESUMO
The diagnosis of "poorly differentiated" carcinoma was made in 47 of 683 colon cancers on the basis of conventional light microscopy which showed poorly defined glands, solid architecture or variable admixtures thereof. Samples from 44 of these 47 tumors were assessed by immunohistochemical analysis for the presence of neuroendocrine (NE) antigens. Paraffin sections were immunostained with antibodies to NSE, chromogranin, serotonin, VIP, substance P and somatostatin. Additional sections were also stained with monoclonal antibody (Mab) A-80 that recognizes a glycoprotein related to exocrine (EX) differentiation. Based on our findings, the tumors were phenotypically reclassified as follows: I) pure EX (n = 8), II) pure NE (n = 4), III) mixed EX-NE carcinomas (n = 23), and IV) predominantly EX carcinomas with occasional NE cells (n = 9). Survival among groups II and III appeared to be less than group I and survival in group IV was significantly less than group I. Survival among the four pure NE (group II) and 11 predominantly NE mixed carcinomas (group III) taken together was significantly less than the pure EX carcinomas. This study indicates: 1) The incidence of NE differentiation in tumors of the colon and rectum is higher than previously believed. 2) The poorly differentiated colon carcinomas comprise four distinct groups: pure EX, pure NE, mixed EX-NE carcinomas, and predominantly EX carcinomas with a NE cell subpopulation. 3) The presence of NE differentiation or of a NE cell subpopulation in colon carcinoma appears to be associated with a poorer prognosis.
Assuntos
Carcinoma/patologia , Neoplasias do Colo/patologia , Proteínas do Tecido Nervoso/análise , Idoso , Idoso de 80 Anos ou mais , Diferenciação Celular , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Estadiamento de NeoplasiasRESUMO
A red fox (Vulpes vulpes) and a European badger (Meles meles) were found dead on a golf-course in October 1997 near Stockholm (Sweden). At necropsy, both animals were obese and the main finding was acute circulatory collapse. Theobromine intoxication was suspected as chocolate waste was available at a nearby farm and no other cause of death could be detected. Gastric contents and samples of liver from both animals were analyzed by reversed-phase high pressure liquid chromatography for the presence of methylxanthines. Theobromine and caffeine were detected in gastric contents and theobromine was identified in the liver samples from both animals. This appears to be the first report of theobromine intoxication in the red fox and the European badger.
Assuntos
Cacau/intoxicação , Carnívoros , Raposas , Teobromina/intoxicação , Doenças dos Animais/induzido quimicamente , Animais , Cafeína/análise , Carnívoros/metabolismo , Cromatografia Líquida de Alta Pressão/veterinária , Evolução Fatal , Feminino , Raposas/metabolismo , Fígado/química , Masculino , Suécia , Xantinas/análiseRESUMO
The aim of the current study was to look for evidence of possible cross-species transmission of Brachyspira species between rodents and farm animals. To do this, previously collected and characterised Brachyspira isolates from rodents, pigs and chickens on the same farms were analysed by random amplified polymorphic DNA (RAPD). Isolates with similar RAPD banding patterns were further typed by pulsed-field gel electrophoresis (PFGE). Identical isolates of Brachyspira pilosicoli, Brachyspira intermedia, Brachyspira murdochii and Brachyspira innocens from pigs and rodents and of B. murdochii from laying hens and rodents were found, indicating cross-species transmission at farm level. PFGE data from rodent isolates of Brachyspira hyodysenteriae were compared with PFGE data from previously typed field isolates of B. hyodysenteriae from pigs with swine dysentery and isolates from mallards (Anas platyrhynchos). Three of four isolates of B. hyodysenteriae from rodents were similar to porcine field isolates by PFGE. PCR analyses of the plasmid-encoded and potential virulence determinants rfb genes B, A, D and C showed that they were present in isolates of B. hyodysenteriae of porcine, mallard and rodent origin.
Assuntos
Brachyspira/classificação , Infecções por Bactérias Gram-Negativas/veterinária , Animais , Brachyspira/genética , Brachyspira/isolamento & purificação , Galinhas , Patos , Eletroforese em Gel de Campo Pulsado/veterinária , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/transmissão , Filogenia , Doenças das Aves Domésticas/microbiologia , Doenças das Aves Domésticas/transmissão , Técnica de Amplificação ao Acaso de DNA Polimórfico , Ratos , Doenças dos Roedores/microbiologia , Sus scrofa , Suínos , Doenças dos Suínos/microbiologia , Doenças dos Suínos/transmissãoAssuntos
Biomarcadores Tumorais/análise , Sistema Nervoso Central , Citoesqueleto/metabolismo , Filamentos Intermediários/metabolismo , Neoplasias Embrionárias de Células Germinativas/metabolismo , Neoplasias do Sistema Nervoso/metabolismo , Humanos , Imuno-Histoquímica , Proteínas de Filamentos Intermediários/metabolismoRESUMO
Brachyspira hyodysenteriae, the aetiological agent of swine dysentery, and a recently proposed and closely related enteropathogenic spirochaete "Brachyspira suanatina", originally isolated from pigs or mallards (Anas platyrhynchos), were used to inoculate week-old mallard ducklings orally or cloacally. The colonization rate, clinical outcome, faecal dry matter content, blood leucocyte counts and gross, microscopical and electron microscopical features 14-16 days post-inoculation were investigated at necropsy examination. Strains of "B. suanatina" of pig and mallard origin and B. hyodysenteriae of mallard origin colonized the ducklings by oral inoculation, and colonization was also established by cloacal inoculation with a "B. suanatina" strain of mallard origin. The porcine reference strain of B. hyodysenteriae (B204) failed to colonize the birds. Unchallenged contact birds in one of the challenge groups were readily colonized by a strain of "B. suanatina" of mallard origin. The proportion of colonized birds differed significantly between the challenge groups (P < 0.0001). For each challenge group, the inoculum and a randomly selected subset of recovered isolates had an identical biochemical profile and banding pattern by randomly amplified polymorphic DNA (RAPD) analysis. None of the birds developed clinical signs of gastrointestinal disease during the trial. The faecal dry weight contents, body weights and total leucocyte and heterophil counts did not differ between the various groups of birds. At the microscopical and electron microscopical levels, the caecal mucosa in some of the Brachyspira culture-positive birds had sharply demarcated epithelial cell changes and there were features of irreversible cell damage in crypt necks coinciding with spirochaetal infiltration of the mucosa. The crypts in Brachyspira culture-positive birds were deeper than in culture-negative birds (median: 237 microm and 218 microm, respectively, P = 0.019). This challenge model was well suited for use in mallards and consistent with previous findings that strongly haemolytic Brachyspira spp. may cross the species barrier between pigs and birds.
Assuntos
Brachyspira/patogenicidade , Infecções por Bactérias Gram-Negativas/veterinária , Doenças dos Suínos/microbiologia , Animais , Brachyspira/genética , Brachyspira/isolamento & purificação , Patos/genética , Patos/microbiologia , Fezes/microbiologia , Mucosa Gástrica/microbiologia , Infecções por Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/microbiologia , Microscopia Eletrônica , Estatísticas não Paramétricas , Suínos/genética , Suínos/microbiologia , Doenças dos Suínos/genéticaRESUMO
The purpose of this study was to evaluate a multilocus sequence typing (MLST) scheme for intestinal spirochaetes of the genus Brachyspira. Eight loci mainly coding for enzymes previously used in multilocus enzyme electrophoresis analysis of Brachyspira species were examined in 66 Brachyspira field isolates and type/reference strains. The isolates and strains were recovered from pigs, birds, dogs and a mouse and originated from seven European countries, the USA and Canada. Forty-six isolates represented recognized Brachyspira species and 20 represented provisionally designated species or isolates that have not been classified. Only two loci gave PCR products for all 66 strains and isolates, but amplicons for seven loci were obtained for 44 of the isolates. Sequences for each locus had a DNA allelic variation of 30-47 and an amino acid allelic variation of 14-47 that gave rise to the same number of sequence and amino acid types (58) for the strains and isolates studied. A population snapshot based on sequence and amino acid types showed a close phylogenetic relationship amongst the porcine isolates from the same geographical regions, and indicated a close evolutionary relationship between isolates recovered from pigs and mallards. A general concordance was obtained between the MLST groupings and classifications based on culture and biochemical tests, 16S rDNA sequence analysis and random amplified polymorphic DNA analysis. This is a first step towards establishing an MLST system for use in identifying Brachyspira species and determining relationships between individual strains and species in the genus.
Assuntos
Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana , Brachyspira/classificação , Intestinos/microbiologia , Análise de Sequência de DNA , Infecções por Spirochaetales/veterinária , Animais , Doenças das Aves/epidemiologia , Doenças das Aves/microbiologia , Aves , Brachyspira/genética , Brachyspira/isolamento & purificação , Doenças do Cão/epidemiologia , Doenças do Cão/microbiologia , Cães , Camundongos , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Infecções por Spirochaetales/microbiologia , Suínos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/microbiologiaAssuntos
Galinhas , Infecções por Herpesviridae/veterinária , Herpesvirus Galináceo 1 , Doenças das Aves Domésticas/prevenção & controle , Vacinação/veterinária , Animais , Surtos de Doenças/veterinária , Feminino , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/prevenção & controle , Infecções por Herpesviridae/transmissão , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/transmissão , Latência Viral , Eliminação de Partículas ViraisRESUMO
The authors studied by immunohistochemistry the intermediate filament (IF) protein profile of 66 frozen samples of breast tissue, including normal parenchyma, all variants of fibrocystic disease (FCD), fibroadenomas, cystosarcoma phylloides, and ductal and lobular carcinomas. Monoclonal antibodies (MAbs) to cytokeratins included MAb KA 1, which binds to polypeptide 5 in a complex with polypeptide 14 and recognizes preferentially myoepithelial cells; MAb KA4, which binds to polypeptides 14, 15, 16 and 19; individual MAbs to polypeptides 7, 13, and 16, 17, 18, and 19, and the MAb mixture AE1/AE3. The authors also applied three MAbs to vimentin (Vim), and three MAbs to glial filament protein (GFP). Selected samples were studied by double-label immunofluorescence microscopy and by staining sequential sections with some of the said MAbs, an MAb to alpha-smooth muscle actin, and well-characterized polyclonal antibodies for the possible coexpression of diverse types of cytoskeletal proteins. Gel electrophoresis and immunoblot analysis also were performed. All samples reacted for cytokeratins with MAbs AE1/AE3, although the reaction did not involve all cells. Monoclonal antibody KA4 stained preferentially the luminal-secretory cells in the normal breast and in FCD, whereas it stained the vast majority of cells in all carcinomas. Monoclonal antibody KA1 stained preferentially the basal-myoepithelial cells of the normal breast and FCD while staining tumor cell subpopulations in 4 of 31 carcinomas. Vimentin-positive cells were found in 8 of 12 normal breasts and in 12 of 20 FCD; in most cases, Vim-reactive cells appeared to be myoepithelial, but occasional luminal cells were also stained. Variable subpopulations of Vim-positive cells were noted in 9 of 20 ductal and in 1 of 7 lobular carcinomas. Glial filament protein-reactive cells were found in normal breast lobules and ducts and in 15 of 20 cases of FCD; with rare exceptions, GFP-reactivity was restricted to basally located, myoepithelial-appearing cells. Occasional GFP-reactive cells were found in 3 of 31 carcinomas. Evaluation of sequential sections and double-label immunofluorescence microscopy showed the coexpression of certain cytokeratins (possibly including polypeptides 14 and 17) with vimentin and alpha-smooth muscle actin together with GFP in some myoepithelial cells. The presence of GFP in myoepithelial cells was confirmed by gel electrophoresis and immunoblotting. Our results indicate that coexpression of cytokeratin with vimentin and/or GFP is comparatively frequent in normal basal-myoepithelial cells of the breast.(ABSTRACT TRUNCATED AT 400 WORDS)