Epitope identification and <Emphasis Type="Italic">in silico</Emphasis> prediction of the specificity of antibodies binding to the coat proteins of <Emphasis Type="Italic">Potato Virus Y</Emphasis> strains

A phage library containing 2.7 × 10⁹ randomly expressed peptides was used to determine the epitopes of three monoclonal antibodies that bind to the coat protein of potato virus Y. Construction of the consensus sequences for the peptides obtained after three selection rounds indicated that each antibody recognized a different epitope located within the first 50 N-terminal amino acids of the coat protein. The location of the epitopes was confirmed by heterologous expression of the N-terminal part of the coat protein in Escherichia coli, and, subsequently, by performing an immunological test with the three antibodies. The accuracy of the phage library was demonstrated by predicting in silico the cross-reactivity of the three antibodies with other potyvirus family members. ELISA and in silico predictions revealed the same results in almost every case. The potential of peptide phage libraries to optimize the use of antibodies in plant virology is discussed.     

Attaching and effacing Escherichia coli isolated from dogs in Brazil: characteristics and serotypic relationship to human enteropathogenic E. coli (EPEC)

Escherichia coli isolates recovered from 182 fecal specimens from dogs up to five months old from the cities of S?o Paulo and Campinas, SP, Brazil, were examined by polymerase chain reaction (PCR) for several virulence factors and properties. The eae gene was found in 23 isolates of E. coli from 22 dogs, 19 of 146 (13%) from dogs with diarrhea and 3 of 36 (8.3%) from dogs with no diarrhea. Two different eae+ isolates were recovered from one dog with diarrhea. Isolates from two dogs with diarrhea harbored the bfpA gene, and none of the isolates possessed genes for enterotoxins, the EAF plasmid or Shiga toxins. PCR showed that, among the 23 isolates, eight were positive for beta intimin, six for gamma, two for, one for alpha, one for kappa, and five showed no amplification with any of the nine pairs of specific intimin primers used. PCR also showed that the LEE (locus of enterocyte effacement) was inserted in selC in four isolates, likely in pheU in seven isolates, and in undetermined sites in twelve isolates. Fifteen isolates adhered to HEp-2 cells and were fluorescence actin staining (FAS) positive. The predominant adherence pattern was the localized adherence-like (LAL) pattern. The eae-positive isolates belonged to a wide diversity of serotypes, including O111:H25, O119:H2 and O142:H6, which are serotypes that are common among human EPEC. These results confirmed the presence of EPEC in dogs (DEPEC) with and without diarrhea. The virulence factors found in these strains were similar to those in human EPEC, leading to the possibility that EPEC may move back and forth among human and canine populations.     

IMAGING DIAGNOSIS–PULMONARY‐TRACHEOBRONCHIAL PROLAPSE IN A NEW CALEDONIAN GIANT GECKO (RHACODACTYLUS LEACHIANUS)

A 3‐year‐old male New Caledonian giant gecko, or Leach's gecko (Rhacodactylus leachianus) presented with acute lethargy and coelomic distention. Findings from survey radiographs and an upper gastrointestinal tract contrast study were consistent with severe aerophagia, a collapsed left lung, and hyperinflation of the right lung due to suspected bronchial obstruction. The gecko was treated with conservative medical management, but was found dead 5 days after presentation. Necropsy findings showed intussusception of the proximal left lung into the left mainstem bronchus and trachea.     

Spirocerca lupi infection in the dog: a review

Spirocercosis is a disease occurring predominantly in Canidae, caused by the nematode Spirocerca lupi. Typical clinical signs are regurgitation, vomiting and dyspnoea. The life-cycle involves an intermediate (coprophagous beetle) and a variety of paratenic hosts. Larvae follow a specific migratory route, penetrating the gastric mucosa of the host, migrating along arteries, maturing in the thoracic aorta before eventually moving to the caudal oesophagus. Here the worm lives in nodules and passes larvated eggs which can be detected using zinc sulphate faecal flotation. Histologically, the mature oesophageal nodule is composed mostly of actively dividing fibroblasts. Spirocerca lupi-associated oesophageal sarcomas may occur and damage to the aorta results in aneurysms. A pathognomonic lesion for spirocercosis is spondylitis of the thoracic vertebrae. Primary radiological lesions include an oesophageal mass, usually in the terminal oesophagus, spondylitis, and undulation of the aortic border. Contrast radiography and computed tomography are helpful additional emerging modalities. Oesophageal endoscopy has a greater diagnostic sensitivity than radiography. Endoscopic biopsies are not sensitive for detecting neoplastic transformation. Doramectin is the current drug of choice, effectively killing adult worms and decreasing egg shedding. Early diagnosis of infection is still a challenge and to date no ideal regimen for prophylaxis has been published.     

Estimating Rainy Season Nitrous Oxide and Methane Fluxes Across Forest and Pasture Landscapes in Costa Rica

The objectives of this research were to estimate exchanges of CH4 and N2O, both radiatively active gases, between soil and atmosphere on hilltop, slope and swale hillslope positions of northeastern Costa Rica; and to assess the importance of accounting for topography in making areal estimates across hilly terrain. Emission rates from soils were measured during the rainy season on three hillslope positions of both actively grazed pastures and primary forests. Emission rates from pasture and forest sites were significantly different for both gases. Differences between slope positions, though notable, were not significantly different for CH4, but were significantly different for N2O. The forest landscape was partitioned with GIS methods into hilltop, slope and swale topographic positions. The calculated areas for each of these were multiplied by their respective emission rates to calculate overall flux from the entire forested area of 618 ha. Nitrous oxide flux ranged from 16,181 to 17,100 g N d-1. Similarly, CH4 flux ranged from -6,201 to -6,658 g CH4d-1. Errors associated with both estimating mean emission rates for each hillslope position and judgmental errors in partitioning the landscape into hillslope positional classes are important to making landscape-scale estimates of flux.