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1.
Digital dermatitis (DD) is a major infectious foot disease of cattle worldwide. Some DD stages are associated with lameness, and the disease has significant economic and animal welfare consequences. The pathogenesis of the disease is not yet fully understood, but Treponema spp. have been associated consistently with clinical cases. Isolation of these fastidious bacteria is difficult and cumbersome. We describe an improved method enabling the culturing of the 3 Treponema spp. (T. pedis, T. phagedenis, and T. medium) from bovine foot specimens derived from DD lesions, using a combination of membrane filtering and subsequent growth on selective agar media. The entire procedure from sampling to verification of individual Treponema spp. takes up to 24 d. In addition, we established a MALDI-TOF MS–based identification method to be applied for confirmation of the different Treponema spp. This scheme provides an unambiguous, simple, and straightforward identification procedure for DD-associated Treponema spp.  相似文献   
2.
BackgroundThe microsporidian parasite Nosema ceranae is a global problem in honeybee populations and is known to cause winter mortality. A sensitive and rapid tool for stable quantitative detection is necessary to establish further research related to the diagnosis, prevention, and treatment of this pathogen.ObjectivesThe present study aimed to develop a quantitative method that incorporates ultra-rapid real-time quantitative polymerase chain reaction (UR-qPCR) for the rapid enumeration of N. ceranae in infected bees.MethodsA procedure for UR-qPCR detection of N. ceranae was developed, and the advantages of molecular detection were evaluated in comparison with microscopic enumeration.ResultsUR-qPCR was more sensitive than microscopic enumeration for detecting two copies of N. ceranae DNA and 24 spores per bee. Meanwhile, the limit of detection by microscopy was 2.40 × 104 spores/bee, and the stable detection level was ≥ 2.40 × 105 spores/bee. The results of N. ceranae calculations from the infected honeybees and purified spores by UR-qPCR showed that the DNA copy number was approximately 8-fold higher than the spore count. Additionally, honeybees infected with N. ceranae with 2.74 × 104 copies of N. ceranae DNA were incapable of detection by microscopy. The results of quantitative analysis using UR-qPCR were accomplished within 20 min.ConclusionsUR-qPCR is expected to be the most rapid molecular method for Nosema detection and has been developed for diagnosing nosemosis at low levels of infection.  相似文献   
3.
Bluetongue (BT) virus serotype 2 (BTV 2) was first confirmed in Tunisia in February 2000 and has since spread northward and westward, infecting several other countries and islands, including Corsica, where clinical disease was reported in October 2000. BT was again reported on the Island in July 2001, some six months after a vaccination campaign against BTV 2. The molecular relationship between isolates of the BTV 2 Corsican wild-type viruses from 2000 and 2001, and the attenuated BTV 2 vaccine were determined by comparing corresponding sequences of genome segments 2, 7 and 10 with each other and with already published sequences available in the genome database. Complete genetic stability was observed between the isolates of the Corsican BTV 2. There was some divergence between the nucleotide sequences of segment 10 obtained from the wild-type and vaccine virus strains. Based on these differences, primers were selected that could be used in RT-PCR to differentiate between the wild-type and the vaccine viruses.  相似文献   
4.
Forty-two horses from Normandy (France) were examined post-mortem for small strongyle infections from October to March. In the positive horses, total worm numbers ranged from 234 to 90,247 (mean 11,297). Encysted larvae represented the major part of the total cyathostome burdens with a high percentage (83%) being early third stage larvae. They were mostly recovered from the caecum (48%) and ventral colon (40%) and were less present in the dorsal colon (12%). Adult cyathostomes were mainly located in the ventral colon (64%) and less frequently in the dorsal colon (27%) and caecum (9%). Twenty species of Cyathostominae were identified. The 10 most prevalent species (in sequence of prevalence) were Cyathostomum coronatum, Cylicocyclus nassatus, Cylicocyclus insigne, Cyathostomum catinatum, Cylicostephanus goldi, Poteriostomum imparidentatum, Cyathostomum labiatum, Cylicocyclus ultrajectinus, Cylicostephanus calicatus and Cylicostephanus minutus which comprised 84% of the total adult population. Twelve species showed a site preference in the ventral colon, five in the dorsal colon and only one in the caecum while two species were collected in nearly equal numbers from the ventral and dorsal colon. The number of species per horse ranged from 1 to 12 with a median of 5. Infections with singletons occurred in 12.5% of the positive horses while multiple infections were encountered in 87.5%. A positive correlation was found between the intensity of cyathostome infection and its diversity, measured either by the number of occurring species or Shanon indexes.  相似文献   
5.
During March 2013, we investigated the presence and the levels of Schmallenberg virus (SBV) circulation in three dairy cow herds and three sheep flocks in Central Macedonia, Greece. In two cow herds, a high number of abortions had been observed during the winter. Six bulk-tank milk samples and 147 individual sera were screened for SBV-specific antibodies by ELISA. Positive reactions were obtained from 5 out of 6 bulk-tank milk samples, 58 out of 90 sera from the 3 cow herds, and 2 sera from 2 of the 3 sheep flocks. Twenty-two ELISA-positive sera were tested by serum neutralization test (SNT). SNT confirmed the presence of neutralizing antibodies against SBV in all samples tested, with titers ranging between 1:32 and ≥1:256. No neutralizing antibodies against Akabane virus (AKAV) or Shamonda virus (SHAV) were detected, indicating that neutralizing antibodies against SBV do not cross react with AKAV or SHAV in SNT. ELISA testing of bulk-tank milk samples proved to be convenient and reliable. None of the tested sera was found positive for SBV by real-time RT-PCR, indicating that the sampling was conducted past the viremia stage. This is the first report of SBV circulation in Greece.  相似文献   
6.
Lipid A is the essential component of endotoxin (Gram-negative lipopolysaccharide), a potent immunostimulatory compound. As the outer surface of the outer membrane, the details of lipid A structure are crucial not only to bacterial pathogenesis but also to membrane integrity. This work characterizes the structure of lipid A in two psychrophiles, Psychromonas marina and Psychrobacter cryohalolentis, and also two mesophiles to which they are related using MALDI-TOF MS and fatty acid methyl ester (FAME) GC-MS. P. marina lipid A is strikingly similar to that of Escherichia coli in organization and total acyl size, but incorporates an unusual doubly unsaturated tetradecadienoyl acyl residue. P. cryohalolentis also shows structural organization similar to a closely related mesophile, Acinetobacter baumannii, however it has generally shorter acyl constituents and shows many acyl variants differing by single methylene (-CH2-) units, a characteristic it shares with the one previously reported psychrotolerant lipid A structure. This work is the first detailed structural characterization of lipid A from an obligate psychrophile and the second from a psychrotolerant species. It reveals distinctive structural features of psychrophilic lipid A in comparison to that of related mesophiles which suggest constitutive adaptations to maintain outer membrane fluidity in cold environments.  相似文献   
7.
Genetically modified (GM) cassava is currently being developed to address problems of diseases that threaten the food security of farmers in developing countries. The technologies are aimed at smallholder farmers, in hopes of reducing the vulnerability of cassava production to these diseases. In this paper we examine barriers to farmers’ voice in the development of GM cassava. We also examine the role of a translational research process to enhance farmers’ voice, to understand the sources of vulnerability farmers in a group in Kenya’s Coast face, and to determine if their concerns are consistent with those of the scientists in agriculture addressing farmers’ needs. A two-way communication participatory process provided insights into the complex vulnerability context of farmers, their primary concerns with processing and markets of cassava in order to improve livelihoods, the lack of networks with two way communication flows, and the lack of information on GM technologies. The translational research engaged farmers and scientists in an iterative process where scientists are learning what farmers need, and farmers are learning about the potential benefits and risks from GM technologies, while at the same time expressing their concerns.  相似文献   
8.
Agroforestry has been suggested to produce an array of ecological and socio-economic benefits that not only reach their adopters, but society as a whole. In spite of strong evidence of the benefits of agroforestry, there is a lack of understanding of the public perceptions of those benefits, which prevents the development of benefit-based promotion strategies of agroforestry products. Thus, this study examines the awareness of these benefits among residents and explores whether perceptions vary across individuals with different socio-demographic characteristics. An on-line platform was used to survey three panels of residents from Missouri, Pennsylvania and Texas. Overall, respondents had a neutral perception of the importance of the ecological and socio-economic benefits that agroforestry farms provide to society when compared to farms employing conventional agricultural practices. These results stress the need to increase public awareness of agroforestry practices and their benefits to more strategically position their products among the final consumer. Gender, education level, and residence location were significantly associated with the perceived ecological benefits produced by agroforestry farms and to a lesser extent with the socio-economic benefits. Results also showed that females and young individuals have a greater awareness of several benefits produced by agroforestry farms. Critical marketing implications to stimulate the purchase of agroforestry products (e.g., berries, nuts) among specific groups of consumers are discussed.  相似文献   
9.
In this article the effect of different nitrogen fertilizer application on the availability of zinc (Zn) content in non calcic brown forest soil was investigated. For this purpose increasing rates of NH4Cl, (NH4)2S04; NH4N03 and Ca(N03)2.4H20 fertilizers were applied for having different physical and chemical properties in two soil samples. An incubation experiment was carried out over 4 months. The available Zn content of the soils was determined each month. According to the results the highest available Zn content of the soils was obtained from NH4Cl applied to the soils followed by (NH4)2S04; NH4N03 and Ca(N03)2.4H20 application to soil. At the end of the experiment the average available Zn content of non calcic brown forest soils for NH4Cl, (NH4)2S04; NH4N03 and Ca(N03)2.4H20 fertilizers are 0.93, 0.91, 0.88 and 0.77 mg kg?1, respectively.  相似文献   
10.

• Introduction  

Wildfires are considered the most important disturbance in the Mediterranean Basin, and some are propagated over long distances due to lift-off and ignition of firebrands.  相似文献   
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