Canine neurogenic Keratoconjunctivitis sicca: 11 cases (2006-2010)

Objective To describe the clinical data of dogs with neurogenic Keratoconjunctivitis sicca (KCS) and an ipsilateral dry nose without other neurologic deficits. Procedure The retrospective case study included 11 dogs diagnosed with neurogenic KCS and an ipsilateral dry nose between 2006 and 2010. Medical records were reviewed for breed, age, sex, history, suspected cause of neurogenic KCS, clinical signs, and treatment modalities. Follow‐up information was obtained by re‐examination of patients or completion of a telephone survey with the referring veterinarian or the owners. Results Mean age of the dogs was 6.6 ± 4.5 years. Neurogenic KCS was diagnosed in three females, five spayed females, one male, and two castrated males representing 10 different breeds. Ophthalmic signs of KCS (mean Schirmer tear test [STT] value of 1.9 ± 2.9 mm/min) combined with an ipsilateral dry nose were diagnosed in seven left and four right eyes. The suspected cause of neurogenic KCS was idiopathic in nine and trauma in two cases. Systemic therapy consisted of oral pilocarpine 1–2% eye drops combined with case‐specific topical treatment with cyclosporine 0.2% and tear substitutes. Duration of systemic treatment with pilocarpine until healing was 125 days (range 84–204, median 98 days) for five dogs. One dog was lost to follow‐up, and the remaining five dogs are still under systemic treatment with pilocarpine. Conclusions Neurogenic KCS with an ipsilateral dry nose seems to be a predominantly idiopathic disease of middle‐aged female dogs without breed predisposition, which may be self‐limiting in some cases.     

Screening for in planta protein-protein interactions combining bimolecular fluorescence complementation with flow cytometry

ABSTRACT: Understanding protein and gene function requires identifying interaction partners usingbiochemical, molecular or genetic tools. In plants, searching for novel protein-proteininteractions is limited to protein purification assays, heterologous in vivo systems such as the yeast-two-hybrid or mutant screens. Ideally one would be able to search for novel proteinpartners in living plant cells. We demonstrate that it is possible to screen for novel proteinproteininteractions from a random library in protoplasted Arabidopsis plant cells and recoversome of the interacting partners. Our screen is based on capturing the bi-molecularcomplementation of mYFP between an YN-bait fusion partner and a completely random preyYC-cDNA library with FACS. The candidate interactions were confirmed using in plantaBiFC assays and in planta FRET-FLIM assays. From this work, we show that the wellcharacterized protein Calcium Dependent Protein Kinase 3 (CPK3) interacts with APX3,HMGB5, ORP2A and a ricin B-related lectin domain containing protein At2g39050. This isone of the first random in planta screens to be successfully employed.     

Ocular manifestations of a metastatic adenocarcinoma in a horse

A 10‐year‐old German Warmblood gelding was referred to the Equine Department of the Vetsuisse Faculty, University of Zurich, Switzerland, for an iris mass OD, lethargy, intermittent fever, and coughing. Ophthalmic examination revealed a 7 × 9 mm raised, fleshy, whitish to pinkish, vascularized iris mass at the 2 o`clock position OD. Fundic examination showed multifocal round, brown to black, slightly raised lesions with indistinct margins and a surrounding hyperreflective zone OU. Physical examination revealed a temperature of 39.2 °C, sinus tachycardia, preputial and ventral edema, and an enlarged right mandibular lymph node. Results of a complete blood count and plasma biochemical profile showed mild anemia, leukocytosis, and thrombocytopenia. Severe splenopathy, moderate splenomegaly, and severe pulmonary pathology with nodules and large areas of consolidated lung parenchyma were observed on abdominal ultrasound and thoracic radiographs, respectively. Fine needle aspirates of the enlarged mandibular lymph node showed malignant epithelial neoplastic cells. The horse was euthanized because of the poor prognosis and subsequently underwent postmortem examination. Macroscopic necropsy and histopathology revealed an adenocarcinoma of suspected pulmonary origin with involvement of eyes, heart, liver, kidneys, spleen, diaphragm, skeletal muscles, mandibular, pulmonary, and internal iliac lymph nodes. Metastatic adenocarcinoma should be considered as a differential diagnosis in horses with iris masses, multifocal chorioretinal infiltrates, and clinical signs that conform to a paraneoplastic syndrome.     

Naturally occurring bacteriophages lyse a large proportion of canine and feline uropathogenic Escherichia coli isolates in vitro

We investigated the feasibility of bacteriophage therapy to combat canine and feline Escherichia coli urinary tract infections (UTIs) by testing the in vitro lytic ability of 40 naturally occurring bacteriophages on 53 uropathogenic E. coli (UPEC). The mean number of UPEC strains lysed by an individual bacteriophage was 21/53 (40%, range 17-72%). In total, 50/53 (94%) of the UPEC strains were killed by one or more of the bacteriophages. Ten bacteriophages lysed 51% of UPEC strains individually and 92% of UPEC strains as a group. Electron microscopy and DNA sequencing of 5 'promising' bacteriophages revealed that 4 bacteriophages belonged to the lytic T4-like genus, while one displayed morphologic similarity to temperate P2-like bacteriophages. Overall, these results indicate that the majority of UPEC are susceptible to lysis by naturally occurring bacteriophages. Thus, bacteriophages show promise as therapeutic agents for treatment of canine and feline E. coli UTIs.     

Haemorrhage caused by thrombocytopenia in 3 calves

The clinical findings and treatment of 3 calves with bleeding attributable to severe thrombocytopenia (1000 to 5000 platelets/microl) are described. The calves responded to treatment with whole blood transfusion, dexamethasone and other drugs. One calf was subsequently diagnosed with persistent bovine viral diarrhoea virus infection and was euthanased despite a favourable response to treatment. The other two calves were discharged after platelet counts had normalised and bleeding had stopped.     

Alternative inoculum sources for fire blight: the potential role of fruit mummies and non‐host plants

Fire blight is the most damaging bacterial disease in apple production worldwide. Cankers and symptomless infected shoots are known as sites for the overwintering of Erwinia amylovora, subsequently providing primary inoculum for infection in the spring. In the present work, further potential sources of inoculum were investigated. Real‐time PCR assays covering a 3‐year‐period classified 19·9% of samples taken from fruit mummies as positive. Bacterial abundance in fruit mummies during autumn, winter and spring was up to 10⁹ cells per gram of tissue and correlated well with later infection rates of blossoms. Blossoms of non‐host plants growing close to infected trees were also shown to be colonized by E. amylovora and to enable epiphytic survival and propagation of bacteria. The results indicate a potential role of fruit mummies and buds in overwintering and as a source of primary inoculum for dissemination of the pathogen early in the growing season. Non‐host blossoms may also serve as an inoculum source in the build‐up of the pathogen population. Both aspects may contribute significantly to the epidemiology of E. amylovora. The significance of infected rootstocks as an inoculum source is also discussed. Fruit mummies might be used to determine pathogen pressure in an orchard before the beginning of the blooming period.     

Isolation,partial identification and application of diazotrophic rhizobacteria from traditional Indian rice cultivars

A diversity of N₂-fixing (diazotrophic) bacteria was isolated from two traditional rice cultivars, Sataria and Kartiki, from the rice growing area of Mithila region of North Bihar, India, where low levels of nitrogen fertilizers are applied. Nitrogen-free semisolid media NFb, JMV and LGI with different carbon sources and pH-values were used for enrichment and isolation of root-associated diazotrophs. The colonization density of roots by diazotrophs, as estimated from positive pellicle formation at highest dilution in nitrogen-free enrichment media, was 10⁶–10⁸ diazotrophic bacteria per g fresh root weight. Roots of the cultivar Kartiki were found to be more densely colonized endophytically by diazotrophs as detected after chloramine T (1%) surface disinfection. To ascertain the phylogenetic affiliation of the isolates, phylogenetic oligonucleotide probes and the Fluorescent in situ Hybridization (FISH) technique were applied. Using group-specific rRNA directed oligonucleotide probes, the majority of the isolates could be identified as alpha-, beta-, or gamma-proteobacteria. Using 16S and 23S rRNA-directed genus- or species-specific probes, Herbaspirillum seropedicae, Azospirillum amazonense, Burkholderia cepacia/vietnamiensis, Rhizobia and Pseudomonas spp. were found to be the most prominent root associated culturable diazotrophs. Diazotrophic Gluconacetobacter spp. were also demonstrated as colonizers of rice roots. Burkholderia cenocepacia, Pseudomonas sp. and three diazotrophic PGPR reference strains were used for the inoculation of axenically grown rice seedlings to determine the plant growth promoting potential. Significant increases in the shoot length (up to 60%), shoot dry weight (up to 33%) and the grain yield (up to 26%) per plant were observed in non-axenic pot and field trials. Using semisolid enrichment media after surface sterilization of field grown inoculated rice roots and oligonucleotide probing of the diazotrophic enrichment cultures, a sustainable colonization with the inoculated bacteria could be demonstrated.     

Isolation and characterisation of equine influenza viruses (H3N8) from Europe and North America from 2008 to 2009

Like other influenza A viruses, equine influenza virus undergoes antigenic drift. It is therefore essential that surveillance is carried out to ensure that recommended strains for inclusion in vaccines are kept up to date. Here we report antigenic and genetic characterisation carried out on equine influenza virus strains isolated in North America and Europe over a 2-year period from 2008 to 2009. Nasopharyngeal swabs were taken from equines showing acute clinical signs and submitted to diagnostic laboratories for testing and virus isolation in eggs. The sequence of the HA1 portion of the viral haemagglutinin was determined for each strain. Where possible, sequence was determined directly from swab material as well as from virus isolated in eggs. In Europe, 20 viruses were isolated from 15 sporadic outbreaks and 5 viruses were isolated from North America. All of the European and North American viruses were characterised as members of the Florida sublineage, with similarity to A/eq/Lincolnshire/1/07 (clade 1) or A/eq/Richmond/1/07 (clade 2). Antigenic characterisation by haemagglutination inhibition assay indicated that the two clades could be readily distinguished and there were also at least seven amino acid differences between them. The selection of vaccine strains for 2010 by the expert surveillance panel have taken these differences into account and it is now recommended that representatives of both Florida clade 1 and clade 2 are included in vaccines.     

GC-based analysis of plant stanyl fatty acid esters in enriched foods

Approaches for the capillary gas chromatographic (GC) based analysis of intact plant stanyl esters in enriched foods were developed. Reference compounds were synthesized by enzyme-catalyzed transesterifications. Their identities were confirmed by means of mass spectrometry. Using a medium polar trifluoropropylmethyl polysiloxane stationary phase, long-chain plant stanyl esters could be separated according to their stanol moieties and their fatty acid chains. Thermal degradation during GC analysis was compensated by determining response factors; calibrations were performed for ten individual plant stanyl esters. For the analysis of low-fat products (skimmed milk drinking yogurts), the GC separation was combined with a "fast extraction" under acidic conditions. For fat-based foods (margarines), online coupled LC-GC offered an elegant and efficient way to avoid time-consuming sample preparation steps. The robust and rapid methods allow conclusions on both, the stanol profiles and the fatty acid moieties, and thus provide a basis for the authentication of this type of functional food ingredients.