Changes in the risk management of Salmonella enterica subspecies diarizonae serovar 61:(k):1, 5, (7) in Swedish sheep herds and sheep meat due to the results of a prevalence study 2012

Background

The prevalence of Salmonella in food producing animals is very low in Sweden due to rigorous control programmes. However, no active surveillance is in place in sheep. The authorities decided to perform a prevalence study in sheep herds because findings at slaughter indicated that sheep associated S. diarizonae (S. enterica subspecies diarizonae serovar 61:(k):1, 5, (7)) might be common in sheep. Sampling was stratified by herd size in two groups, small herds with ≤ 30 animals and large herds with > 30 animals. In each stratum, 237 herds were selected at random. Faecal samples received from 244 out of the 474 randomly selected herds were analysed.

Results

A total of 40 of 100 (40%) of large herds and 17 of 144 (12%) of small herds were positive. The overall adjusted prevalence was 17.6% (95% CI, 12.9-22.2). Sheep associated S. diarizonae was detected in all counties (n = 21). Scientific opinions and an evaluation of on-farm control measures performed concluded that the impact of sheep associated S. diarizonae on human health is very low, and that risk management measures applied in response to findings of sheep associated S. diarizonae in sheep or sheep meat can be expected to have very little impact on reducing risks to human health. As a result, Swedish authorities decided to make an exemption for sheep associated Salmonella diarizonae in sheep and sheep meat in the current Salmonella control measures.

Conclusions

Sheep associated S. diarizonae is endemic in Swedish sheep herds. It is more common in large herds and not limited to certain parts of the country. The responsible authorities concluded that current risk management actions regarding sheep associated S. diarizonae in sheep and sheep meat are not proportional to the risk. This is the first time in the history of the Swedish Salmonella control programme that an exemption from the legislation has been made for a specific serovar. If there is any future indication of an increasing risk, due to e.g. change in the pathogenicity or development of antimicrobial resistance, the risk assessment will be re-evaluated and control measures reinforced if needed.     

Diabetes mellitus in a population of 180,000 insured dogs: incidence, survival, and breed distribution

BACKGROUND: Canine diabetes mellitus (DM) is a common endocrinopathy with an unclear etiology. For a better understanding of the underlying mechanisms, there is a need for comprehensive epidemiologic studies. Earlier studies have shown that the risk of disease is higher in certain dog breeds. HYPOTHESIS: Incidence, age of onset, survival and sex proportion of DM vary by breed. ANIMALS: Data from a cohort of 182,087 insured dogs aged 5-12 years accounting for 652,898 dog-years at risk were studied retrospectively. METHODS: Incidence rates by sex, breed, and geography were calculated with exact denominators. Age-specific incidence and survival after 1st DM claim were computed with Cox's regression and Kaplan-Meier survival function. Multivariable survival analysis was performed for the outcome diagnosis of DM with age, sex, and geography tested as fixed effects, previous endocrine or pancreatic diseases tested as time-dependent covariates, and breed tested as a random effect. RESULTS: The mean age at 1st insurance claim for the 860 DM dogs (72% females) was 8.6 years. The incidence of DM was 13 cases per 10,000 dog-years at risk. Australian Terriers, Samoyeds, Swedish Elkhounds, and Swedish Lapphunds were found to have the highest incidence. The proportion of females with DM varied significantly among breeds. Swedish Elkhounds, Beagles, Norwegian Elkhounds, and Border Collies that developed DM were almost exclusively females. The multivariable model showed that breed, previous hyperadrenocorticism, and female sex were risk factors for developing DM. Median survival time was 57 days after 1st claim. Excluding the 223 dogs that died within 1 day, the median survival time was 2 years after 1st claim of DM. CONCLUSION: The significant breed-specific sex and age differences shown in this study indicate that genetic variation could make breeds more or less susceptible to different types of DM.     

Combining information from surveys of several species to estimate the probability of freedom from Echinococcus multilocularis in Sweden,Finland and mainland Norway

Background

The fox tapeworm Echinococcus multilocularis has foxes and other canids as definitive host and rodents as intermediate hosts. However, most mammals can be accidental intermediate hosts and the larval stage may cause serious disease in humans. The parasite has never been detected in Sweden, Finland and mainland Norway. All three countries require currently an anthelminthic treatment for dogs and cats prior to entry in order to prevent introduction of the parasite. Documentation of freedom from E. multilocularis is necessary for justification of the present import requirements.

Methods

The probability that Sweden, Finland and mainland Norway were free from E. multilocularis and the sensitivity of the surveillance systems were estimated using scenario trees. Surveillance data from five animal species were included in the study: red fox (Vulpes vulpes), raccoon dog (Nyctereutes procyonoides), domestic pig, wild boar (Sus scrofa) and voles and lemmings (Arvicolinae).

Results

The cumulative probability of freedom from EM in December 2009 was high in all three countries, 0.98 (95% CI 0.96-0.99) in Finland and 0.99 (0.97-0.995) in Sweden and 0.98 (0.95-0.99) in Norway.

Conclusions

Results from the model confirm that there is a high probability that in 2009 the countries were free from E. multilocularis. The sensitivity analyses showed that the choice of the design prevalences in different infected populations was influential. Therefore more knowledge on expected prevalences for E. multilocularis in infected populations of different species is desirable to reduce residual uncertainty of the results.     

Poinsettia protoplasts - a simple, robust and efficient system for transient gene expression studies

ABSTRACT: BACKGROUND: Transient gene expression systems are indispensable tools in molecular biology. Yet, their routine application is limited to only few plant species and often requires substantial equipment and facilities. The high content of chloroplasts and chlorophyll can impede downstream applications of transformed cells from green plant tissue. RESULTS: We describe a fast and simple technique for the high-yield isolation and efficient transformation (>70%) of mesophyll-derived protoplasts from red leaves of the perennial plant Poinsettia (Euphorbia pulccherrima) for which no particular growth facilities or expensive equipment are needed. Poinsettia protoplasts display an astonishing robustness and can be employed in all commonly-used downstream applications, such as subcellular localisation (multi-colour fluorescence) or promoter activity studies. Due to a low abundance of chloroplasts or chromoplasts, problems encountered in other mesophyll-derived protoplast systems (particularly autofluorescence) are alleviated. Transgene expression is detectable within 90 min post-transformation and lasts over several days. CONCLUSIONS: The simplicity of isolation and transformation renders Poinsettia protoplasts an attractive system for transient gene expression experiments, including multicolour fluorescence, subcellular localisation and promoter activity studies. In addition, they offer hitherto unknown possibilities for anthocyan research and industrial applications.     

Landscape contrast: a solution to hidden assumptions in the metacommunity concept?

The metacommunity concept provides a spatial perspective on community dynamics, and the landscape provides the physical template for a metacommunity. Several aspects of landscape heterogeneity, such as landscape diversity and composition, and characteristics of the matrix between habitat patches such as habitat connectivity, and geometry of habitat patches, may moderate metacommunity processes. These aspects of landscape heterogeneity are rarely considered explicitly in the metacommunity discussion, however. We propose landscape contrast (i.e., the average dissimilarity in habitat quality between neighboring patches) as a key dimension of landscape heterogeneity. The concept of landscape contrast unifies discrete and continuous landscape representations (homogeneous, gradient, mosaic and binary) and offers a means to integrate landscape heterogeneity in the metacommunity concept. Landscape contrast as perceived by the organisms affects several fundamental metacommunity processes and may thus constrain which metacommunity models may be observed. In a review of empirical metacommunity studies (n = 123), only 22 % of studies were explicit about their underlying landscape model assumptions, with striking differences among taxonomic groups. The assumed landscape model constrained, but did not determine, metacommunity models. Integration and explicit investigation of landscape contrast effects in metacommunity studies are likely to advance ecological theory and facilitate its application to real-world conservation problems.     

Site fidelity and movements of an amphidromous goby revealed by otolith multi‐elemental signatures along a tropical watershed

Otolith microchemistry can provide crucial information to address gaps of knowledge in spatio‐temporal ecology of fish species. However, understanding the seasonal variability of water chemistry and its effect on otolith signatures is needed to interpret fish movements. Otolith multi‐elemental signatures were used to examine the diadromous migration and small‐scale movements of a tropical goby (Sicyopterus lagocephalus) within a watershed in La Réunion Island, Indian Ocean. The elemental ratios Sr:Ca, Ba:Ca and Sr:Ba recorded monthly in the water of nine sampling sites varied between localities and were correlated with otolith signatures, whereas the Mg:Ca signature was not. The incorporation of some trace elements in otoliths was influenced by the size of fish and site where they were caught, reflecting an effect of endogenous and exogenous factors. Despite these sources of variation, the elemental signatures did not overlap for three river sections distributed along the 26‐km‐long watershed. The diadromous migration and fish lifetime movements between these river sections were reconstructed using an original process based on random forest analysis. This approach revealed distinct migratory behaviours after fish entered in the river, with some individuals progressing slowly in the watershed whereas other rapidly reached the upper localities. As adults, S. lagocephalus showed a restricted exploratory behaviour and a strong site fidelity to a particular part of the watershed. The detection of adult movement was typically only between adjacent locations and is possibly explained by shifts in local intraspecific hierarchies and/or displacement during extreme discharge caused by cyclones.     

Site-Directed Disruption of the fimA and fimF Fimbrial Genes of Xylella fastidiosa

ABSTRACT Xylella fastidiosa causes Pierce's disease, a serious disease of grape, citrus variegated chlorosis, almond and oleander leaf scorches, and many other similar diseases. Although the complete genome sequences of several strains of this organism are now available, the function of most genes in this organism, especially those conferring virulence, is lacking. Attachment of X. fastidiosa to xylem vessels and insect vectors may be required for virulence and transmission; therefore, we disrupted fimA and fimF, genes encoding the major fimbrial protein FimA and a homolog of the fimbrial adhesin MrkD, to determine their role in the attachment process. Disruption of the fimA and fimF genes in Temecula1 and STL grape strains of X. fastidiosa was obtained by homologous recombination using plasmids pFAK and pFFK, respectively. These vectors contained a kanamycin resistance gene cloned into either the fimA or fimF genes of X. fastidiosa grape strains Temecula1 or STL. Efficiency of transformation was sufficiently high ( approximately 600 transformants per mug of pFFK DNA) to enable selection of rare recombination events. Polymerase chain reaction and Southern blot analyses of the mutants indicated that a double crossover event had occurred exclusively within the fimA and fimF genes, replacing the chromosomal gene with the disrupted gene and abolishing production of the corresponding proteins, FimA or FimF. Scanning electron microscopy revealed that fimbriae size and number, cell aggregation, and cell size were reduced for the FimA or FimF mutants of X. fastidiosa when compared with the parental strain. FimA or FimF mutants of X. fastidiosa remained pathogenic to grapevines, with bacterial populations slightly reduced compared with those of the wild-type X. fastidiosa cells. These mutants maintained their resistance to kanamycin in planta for at least 6 months in the greenhouse.     

Impact of hardness of model fresh cheese on aroma release: in vivo and in vitro study

Using atmospheric pressure chemical ionization-mass spectrometry, aroma release was investigated in vivo and in vitro from three cheese-like gels with different hardnesses. In vivo, nosespace experiments were performed with 14 subjects. Results showed that the harder gel induced a greater and a faster release of all aroma compounds. In vitro, aroma release was followed in a mouth simulator where breakdown was mechanically produced. The same rate of stirring was applied to the three gels. In these conditions, we found that the amount of aroma released from the three gels was not discriminant. Thus, modification of gel structure had a strong impact on in vivo aroma release, but structural variations alone were not sufficient to induce a greater release. Natural breakdown provided by panelists during food consumption and adapted to the texture of the food was proposed to be the key parameter affecting in vivo aroma release.