Canine leishmaniosis in Central Europe: retrospective survey and serological study of imported and travelling dogs

Canine leishmaniosis is a common parasitic disease in Central Europe affecting dogs imported or returning from endemic countries around the Mediterranean basin. Through an internet discussion forum owners of dogs with suspected or proven leishmaniosis living in Central Europe (D, A, CH), were questioned about the dog's history. Additionally, serologic examinations of the dogs for anti-Leishmania antibodies (ELISA using antigen of promastigote stages) were offered to the participants. From February to October 2003, 291 dogs imported or returning from Southern Europe (Spain, Italy, Greece, Turkey, France, Malta, Portugal and others) were analysed. Serologically, 111 dogs (38%) were classified positive; 103 being imported and eight travelling dogs. The majority of seropositive dogs originated from Spain (67%). No significant correlation could be established between race, sex and age and the incidence of the disease. The clinical symptoms in the seropositive dogs varied widely and ranged from mild general symptoms to visceral manifestations with chronic renal failure. Skin disorders were found in 78% of the seropositive, symptomatic animals. In the country of origin or after import or return, 174 out of 291 dogs (60%) had been tested for the presence of anti-Leishmania antibodies by different immunofluorescence antibody tests (IFAT). Discrepancies between the ELISA and the various IFATs used were noted in 55 cases (32%), especially in cases of low IFAT titers. Most of the seropositive dogs (80%) had been treated against leishmaniosis. In 91% of these cases, Allopurinol as monotherapy or in combination with Glucantime had been used. For diagnostics and therapy, dog owners had spent an average of 1,100 euros (median 900 euros, maximum 5,800 euros).     

Maedi-visna impact on productivity in Quebec sheep flocks (Canada)

An epidemiological study was conducted to determine the impact of maedi-visna (MV) seropositivity on productivity in commercial sheep flocks of the province of Quebec, Canada. A total of 1734 ewes and 220 rams were selected randomly from 29 flocks distributed in the Bas-St-Laurent and Estrie regions. Serostatus was determined with an enzyme-linked immunosorbent assay (ELISA) using recombinant proteins.Flock-specific, animal-level seroprevalence varied from 3 to 70% (median=29%). Seroprevalence increased with age and size of the flock, and was higher in ewes relative to rams (but was not associated with body score). A decrease of 0.94 kg per lamb in weaning weight was seen only for lambs raised by seropositive ewes >/=4 years old, and seropositivity in ewes of any age was associated with an increase in 0-30 days lamb-mortality (OR: 1.65). The impact of MV infection on weaning weight and lamb mortality did not vary between flocks, and seropositivity in ewes was not associated with litter size or lamb's birth weight.     

Species and strain specific identification of lactic acid bacteria in complex microflora

The identification of lactic acid bacteria in a complex microbiota using bacteriological culture in combination with phenotypic and genotypic identification techniques is laborious and time-consuming. New molecular methods permit a fast and culture-independent characterisation of such microbiota. Denaturing gradient gel electrophoresis (DGGE) of PCR fragments of the 16S rRNA gene has been proven to be a suitable tool. Here the use of PCR-DGGE with group specific primers is described to investigate the dynamic of sourdough microbiota from addition of the starter until the microbiota remained stable. Species were identified by applying an identification ladder obtained from reference strains or by sequence analysis of the PCR fragments. Furthermore, a method for detection of strains in complex microbiota is described. A strain specific chromosomal DNA fragment of Lactobacillus paracasei LTH 2579 was isolated applying the subtraction hybridisation. Based on the acquired target sequence a specific PCR system was established and combined with a PCR system specific for the species L. paracasei. Use of this detection system permitted to identify and quantitatively detect L. paracasei LTH 2579 in fermented sausages and upon consumption in faecal samples.     

Rib osteoblastic osteosarcoma in an African hedgehog (Atelerix albiventris).

A 3-year-old African hedgehog (Atelerix albiventris) was presented to the Exotic Animal Clinic of the University of Montreal for evaluation of a mass growing on the right thoracic wall. The diagnostic workup, which included helical computed tomography, confirmed the presence of a large mass, originating from the right 7th rib, infiltrating the thoracic wall and cavity. The animal was euthanized due to the poor prognosis. At necropsy, a well-demarcated mass penetrated the thoracic wall and incorporated the 6th to 8th ribs. Cut sections of the tumor were white, glistening, firm, and gritty. Microscopically, it was composed of polyhedral to elongated cells with interspersed trabeculae of osteoid and large areas of coagulative necrosis. On the basis of histopathologic findings, a diagnosis of osteoblastic osteosarcoma was made. To the authors' knowledge, this is the first report of an osteoblastic osteosarcoma on the thoracic wall of an African hedgehog, as well as the first report of the use of helical computed tomography in that species.     

Evolutionary history of Salmonella typhi

For microbial pathogens, phylogeographic differentiation seems to be relatively common. However, the neutral population structure of Salmonella enterica serovar Typhi reflects the continued existence of ubiquitous haplotypes over millennia. In contrast, clinical use of fluoroquinolones has yielded at least 15 independent gyrA mutations within a decade and stimulated clonal expansion of haplotype H58 in Asia and Africa. Yet, antibiotic-sensitive strains and haplotypes other than H58 still persist despite selection for antibiotic resistance. Neutral evolution in Typhi appears to reflect the asymptomatic carrier state, and adaptive evolution depends on the rapid transmission of phenotypic changes through acute infections.     

Members of the Kunitz-type protease inhibitor gene family of potato inhibit soluble tuber invertase in vitro

Summary A soluble proteinaceous invertase inhibitor was purified from tubers of cv. Provita. N-terminal amino acid sequencing of the purified protein indicated that the invertase inhibitor was a member of a family of small tuber proteins known as Kunitz-type protease inhibitors. The purified protein was completely inhibitory to soluble tuber invertase at quantities that did not inhibit trypsin. Based on amino acid sequence information of invertase inhibitor protein, seven candidate cDNA clones were identified in libraries of Provita and Saturna tubers. The candidate cDNA sequences differed from each other between 2% and 5%, having several non-conservative amino acid substitutions when compared with sequence related protease inhibitors. The results suggest invertase as an alternative target of tuber “protease” inhibitors.     

Vaccine-induced antibodies linked to bovine neonatal pancytopenia (BNP) recognize cattle major histocompatibility complex class I (MHC I)

ABSTRACT: A mysterious disease affecting calves, named bovine neonatal pancytopenia (BNP), emerged in 2007 in several European countries. Epidemiological studies revealed a connection between BNP and vaccination with an inactivated vaccine against bovine virus diarrhea (BVD). Alloantibodies reacting with blood leukocytes of calves were detected in serum and colostrum of dams, which have given birth to calves affected by BNP. To understand the linkage between vaccination and the development of alloantibodies, we determined the antigens reacting with these alloantibodies. Immunoprecipitation of surface proteins from bovine leukocytes and kidney cells using sera from dams with a confirmed case of BNP in their gestation history reacted with two dominant protein species of 44 and 12 kDa. These proteins were not detected by sera from dams, free of BVDV and not vaccinated against BVD, and from sera of animals vaccinated with a different inactivated BVD vaccine. The 44 kDa protein was identified by mass spectrometry analysis as MHC I, the other as β-2-microglobulin. The presence of major histocompatibility complex class I (MHC I) in the vaccine was confirmed by Western blot using a MHC I specific monoclonal antibody. A model of BNP pathogenesis is proposed.