Genetic diversity in seven populations of Nicaraguan teosinte (<Emphasis Type="Italic">Zea nicaraguensis</Emphasis> Iltis et Benz) as estimated by microsatellite variation

Teosintes are the closest relatives to modern maize, Zea mays L. ssp. mays. They are wild grasses with a native distribution area from Mexico to Nicaragua and represent an important genetic resource. However, the genetic diversity of Nicaraguan teosinte (Zea nicaraguensis Iltis et Benz) has not yet been determined. This teosinte species has decreased in the last 25 years and now must be regarded as an endangered species. An analysis of the genetic diversity of Zea nicaraguensis was carried out in a total of 240 individuals from seven populations. Eleven Simple Sequences Repeat (SSR) primer pairs were used. A total of 42 alleles were found, the range of alleles per locus was 2–5 (mean 3.8) and the numbers of genotypes varied between primers. The primer Bnlg 1538 showed the highest value, with 45 genotypes through all populations. The genetic diversity observed (Ho) between all populations varied from 0.51 to 0.63, with an average of 0.563. One of the populations had as many as 40 alleles. Analysis of molecular variance (AMOVA) revealed that most of the variation was within population, at a significantly high level (P < 0.001). Rare alleles were detected in all populations, but unique alleles were only found in four populations. These results are highly relevant when developing conservation strategies and show that preserving populations in their natural habits is highly important.     

Malignant transformation of a unicameral bone cyst in a cat

A unicameral bone cyst in the proximal humerus of a 3-year-old Norwegian forest cat was diagnosed by dynamic contrast-enhanced magnetic resonance imaging, surgical exploration, and histopathology. Surgical curettage and incorporation of bone cement led to full recovery. An osteosarcoma developed at the surgical site 17 months later. Thoracic radiographs showed pulmonary lesions consistent with metastasis.     

A longitudinal study of seroprevalence and seroconversion of Neospora caninum infection in dairy cattle in northeast Thailand

A long-term study was carried out in 11 dairy herds in the Khon Kaen province of northeast Thailand between August 2001 and November 2004. The objective was to investigate seroprevalence dynamics of Neospora caninum infection in the herds and to demonstrate patterns of seroconversion in individual cattle. Each herd was visited once a year, in total four times, and sera from cattle > 3 months of age and farm dogs as well as a sample from the bulk milk were collected. All samples were analysed for presence of specific antibodies by an N. caninum iscom ELISA. The overall percentage of antibody-positive cattle was constant and varied only between 10 and 13% over the 4 years, but the variation in within-herd seroprevalence between herds was substantial. Two herds had > or = 20% seropositive animals at all samplings and consistently high bulk milk OD, whereas two herds had no seropositive animal at the last two samplings and low bulk milk OD. Five herds had a decreasing trend of within-herd seroprevalence, whereas the remaining six herds had a higher portion of test-positive individuals at the end of the study. A total of 424 individuals were sampled more than once; 344 (81%) and 32 (8%) were consistently antibody-negative and antibody-positive, respectively. The proportions of animals that changed from being seronegative to seropositive and from being seropositive to seronegative between the years were 3.9-4.6% and 19-39%, respectively. Apparent vertical and horizontal transmission rates were 58% (95% CI; 44-71%) and 5% (95% CI; 3-7%), respectively. In conclusion, the overall percentage of N. caninum antibody-positive cattle was constant over the years, but the within-herd seroprevalence varied substantially between the herds. Seroconversions were likely to occur in individual cattle although most animals had consistent serological status throughout the study.     

Genetic relatedness and netB prevalence among environmental Clostridium perfringens strains associated with a broiler flock affected by mild necrotic enteritis

In a previous study we investigated pulsed-field gel electrophoresis (PFGE) genotype diversity and prevalence of the netB toxin gene in Clostridium perfringens (CP) isolates recovered from a broiler flock (flock 1) affected by necrotic enteritis (NE). In this follow-up work, we examined samples collected before placement of flock 1, to see if NE during rearing could be traced back to the cleaned and empty building or the day-old chicks. Litter from the next flock in the same building (flock 2) was also examined. We detected 25 different PFGE genotypes, five of which were found only in litter from flock 2. Six genotypes which had been found in flock 1 during rearing were detected in samples collected before placement. NetB positive isolates belonging to two of these genotypes had been recovered from NE lesions during rearing, suggesting that virulent strains were transmitted from the cleaned and disinfected broiler house. NetB frequency among isolates from the empty building was 45%, indicating that netB positive strains were prevalent in a building that previously had housed a healthy flock offered in-feed narasin (flock 0). NetB frequency among isolates from litter used by flock 2 was 22%, indicating that netB positive strains were present in the environment of a 14-days-old healthy flock offered in-feed narasin. Two prevalent genotypes were consistently either netB negative or netB positive. However, the presence of genotypes represented by both negative and positive isolates may suggest that the gene can spread horizontally among different CP strains.     

Chlorogenic acid,a metabolite identified by untargeted metabolome analysis in resistant tomatoes,inhibits the colonization by Alternaria alternata by inhibiting alternariol biosynthesis

Tomato fruits can be contaminated by saprophytic strains of Alternaria alternata which is the reason for the frequent occurrence of Alternaria toxins like alternariol, alternariol monomethylether or tenuazonic acid in these types of products. It was shown earlier that alternariol is a colonization factor for tomatoes. In the current analysis two different tomato genotypes were analysed by untargeted comprehensive two-dimensional gas chromatography mass spectrometry (GC×GC-MS). This analysis revealed clear differences in the metabolic profiles which were paralleled by differences in resistance towards Alternaria colonization. One of the genotypes was more resistant against A. alternata infection and contained high amounts of chlorogenic acid in contrast to the other genotype which was sensitive against infection. In in vitro analysis, chlorogenic acid reduced alternariol biosynthesis during the first days of growth of A. alternata. Expression analysis of the alternariol polyketide synthase gene, a key gene in the biosynthesis of alternariol, also revealed a temporal reduction in its expression in the first phases of growth. However by chromatographic analysis it could be demonstrated that chlorogenic acid was degraded over time. This degradation leads to a relief of inhibition resulting in an only temporal inhibition of alternariol biosynthesis. In vivo colonization experiments revealed that chlorogenic acid reduces colonization of tomatoes by A. alternata in a concentration dependent manner, which however is partly counteracted by the addition of alterariol.     

Genetic basis of penicillin resistance of S. aureus isolated in bovine mastitis

Background

The blaZ gene encoding penicillin resistance can be located either chromosomally or on plasmids. The aim of this study was to investigate the genetic relationships and to determine the location of the blaZ gene in S. aureus isolated in bovine mastitis in Finland and Sweden.

Methods

Seventy-eight β-lactamase positive S. aureus isolates from bovine mastitis (34 from Finland and 44 from Sweden) were included in the study. The localization of blaZ gene was determined by Southern blotting. The blaZ genes of the isolates were sequenced and the sequences were translated to beta-lactamase proteins and further grouped as different protein signatures. The isolates and, as control, 33 Swedish and 36 Finnish beta-lactamase negative isolates were typed with pulsed-field gel electrophoresis (PFGE).

Results

In 26 out of 34 Finnish isolates (76.5%) and in 25 out of 44 Swedish isolates (56.8%) the blaZ gene was localized on a plasmid. Six different protein signatures were found. One signature was found only in four Swedish isolates, but all other signatures were found both in Finnish and Swedish isolates. The PFGE results revealed a diversity of S. aureus clones. The protein signatures were not clearly associated with certain pulsotypes.

Conclusions

The plasmid location of the blaZ gene was not statistically significantly more common in Finland than in Sweden, and hence does not explain the higher proportion of penicillin-resistant isolates of S. aureus causing bovine mastitis in Finland compared to Sweden.     

Investigations of barley stripe mosaic virus as a gene silencing vector in barley roots and in <Emphasis Type="Italic">Brachypodium distachyon</Emphasis> and oat

Background  

Gene silencing vectors based on Barley stripe mosaic virus (BSMV) are used extensively in cereals to study gene function, but nearly all studies have been limited to genes expressed in leaves of barley and wheat. However since many important aspects of plant biology are based on root-expressed genes we wanted to explore the potential of BSMV for silencing genes in root tissues. Furthermore, the newly completed genome sequence of the emerging cereal model species Brachypodium distachyon as well as the increasing amount of EST sequence information available for oat (Avena species) have created a need for tools to study gene function in these species.