Molecular method for the characterization of Coxiella burnetii from clinical and environmental samples: variability of genotypes in Spain

Background: Coxiella burnetii is a highly clonal microorganism which is difficult to culture, requiring BSL3 conditions for its propagation. This leads to a scarce availability of isolates worldwide. On the other hand, published methods of characterization have delineated up to 8 different genomic groups and 36 genotypes. However, all these methodologies, with the exception of one that exhibited limited discriminatory power (3 genotypes), rely on performing between 10 and 20 PCR amplifications or sequencing long fragments of DNA, which make their direct application to clinical samples impracticable and leads to a scarce accessibility of data on the circulation of C. burnetii genotypes. Results: To assess the variability of this organism in Spain, we have developed a novel method that consists of a multiplex (8 targets) PCR and hybridization with specific probes that reproduce the previous classification of this organism into 8 genomic groups, and up to 16 genotypes. It allows for a direct haracterization from clinical and environmental samples in a single run, which will help in the study of the different genotypes circulating in wild and domestic cycles as well as from sporadic human cases and outbreaks. The method has been validated with reference isolates. A high variability of C. burnetii has been found in Spain among 90 samples tested, detecting 10 different genotypes, being those adaA negative associated with acute Q fever cases presenting as fever of intermediate duration with liver involvement and with chronic cases. Genotypes infecting humans are also found in sheep, goats, rats, wild boar and ticks, and the only genotype found in cattle has never been found among our clinical samples. Conclusions: This newly developed methodology has permitted to demonstrate that C. burnetii is highly variable in Spain. With the data presented here, cattle seem not to participate in the transmission of C. burnetii to humans in the samples studied, while sheep, goats, wild boar, rats and ticks share genotypes with the human population.

L'aprofitament ramader a la vall del Madriu-Perafita-Claror (Principat d'Andorra)

The farming exploitation in the Madriu-Perafita-Claror Valley (Principality of Andorra). The Madriu-Perafita-Claror Valley, a natural space located in the Principality of Andorra, has kept a high ecological and landscape value through time. At present, the Valley is considered in the cultural landscape category of the UNESCO World-wide Heritage. A study of the spatial variability of pastures in the Valley conducted from 1994 to 2003 concluded that there was an optimistic future for livestock. This future was mainly explained by new policies in the country, as well as by the new hopes of the tockbreeders. The study also stated that cattle and horse movements within the Valley did not varied over the study period, although entrance and exit points changed. Sheep only fed in the Madriu-Perafita-Claror Valley, but it wouldbe convenient its introduction in other areas where horses and cattle did not pasture. The study concluded that the use of the Valley by the stockbreeders contributed to the development of the vegetation and the landscape, and that the livestock is very important to keep natural and landscape values of the Valley.

Animals in an Urban Context. A Zooarchaeological study of the Medieval and Post-Medieval town of Turku

The aim of this study is to explore the role and importance of different animal species in Turku through an analysis of osteological data and documentary evidence. The osteological material used in this study is derived from two town plots in Turku dating from the 13th century to the 19th century. The osteological material deposited in Turku represents animals bred both in the town and in the surrounding landscape. Animal husbandry in SW-Finland can also be examined through a number of historical documents. The importance of animals in Turku and its hinterland are closely connected and therefore the roles of the animals in both urban and rural settings are examined. The study has revealed the complexity of the depositional patterns in medieval and post-medieval Turku. In the different areas of Turku, characteristic patterns in the osteological material and different deposit types were evident. These patterns are reflections of the activities and therefore of the lifestyles practiced in Turku. The results emphasise the importance of context- awareness in the study of material culture from archaeological sites. Both the zooarchaeological and historical sources indicate that cattle were important in animal husbandry in Turku from the Middle Ages up to the 19th century. Sheep were the second most common species. When taking into consideration the larger size of cattle, the dominance of these animals when it come to meat consumption seems clear even in those phases where sheep bones are more abundant. Pig is less abundant in the material than either cattle or sheep and their importance for subsistence was probably fairly modest, albeit constant. Goats were not abundant in the material. Most of the identified goat bones came from low utility body parts (e.g. skulls and lower extremities), but some amount of goat meat was also consumed. Wild species were of minor importance when it came to consumption practices in Turku. The changes in Turku’s animal husbandry patterns between the medieval and post medieval periods is reflected in the change in age of the animals slaughtered, which was part of a wider pattern seen in North- and Central Europe. More mature animals are also present in the assemblages. This pattern is related to the more pronounced importance of cattle as a manure producer and a draught animal as a result of the intensification of crop cultivation. This change seems to occur later in Finland than in the more Southerly regions, and indeed it did not necessarily take hold in all parts of the country.

Animal modelling for inherited central vision loss.

Disease-causing variants of a large number of genes trigger inherited retinal degeneration leading to photoreceptor loss. Because cones are essential for daylight and central vision such as reading, mobility, and face recognition, this review focuses on a variety of animal models for cone diseases. The pertinence of using these models to reveal genotype/phenotype correlations and to evaluate new therapeutic strategies is discussed. Interestingly, several large animal models recapitulate human diseases and can serve as a strong base from which to study the biology of disease and to assess the scale-up of new therapies. Examples of innovative approaches will be presented such as lentiviral-based transgenesis in pigs and adeno-associated virus (AAV)-gene transfer into the monkey eye to investigate the neural circuitry plasticity of the visual system. The models reported herein permit the exploration of common mechanisms that exist between different species and the identification and highlighting of pathways that may be specific to primates, including humans.

Bartonella apis sp. nov., a honey bee gut symbiont of the class Alphaproteobacteria.

Here, we report the culture and characterization of an alphaproteobacterium of the order Rhizobiales, isolated from the gut of the honey bee Apis mellifera. Strain PEB0122T shares >95 % 16S rRNA gene sequence similarity with species of the genus Bartonella, a group of mammalian pathogens transmitted by bloodsucking arthropods. Phylogenetic analyses showed that PEB0122T and related strains from the honey bee gut form a sister clade of the genus Bartonella. Optimal growth of strain PEB0122T was obtained on solid media supplemented with defibrinated sheep blood under microaerophilic conditions at 35-37 °C, which is consistent with the cultural characteristics of other species of the genus Bartonella. Reduced growth of strain PEB0122T also occurred under aerobic conditions. The rod-shaped cells of strain PEB0122T had a mean length of 1.2-1.8 μm and revealed hairy surface structures. Strain PEB0122T was positive for catalase, cytochrome c oxidase, urease and nitrate reductase. The fatty acid composition was comparable to those of other species of the genus Bartonella, with palmitic acid (C16 : 0) and isomers of 18- and 19-carbon chains being the most abundant. The genomic DNA G+C content of PEB0122T was determined to be about 45.5 mol%. The high 16S rRNA gene sequence similarity with species of Bartonella and its close phylogenetic position suggest that strain PEB0122T represents a novel species within the genus Bartonella, for which we propose the name Bartonella apis sp. nov. The type strain is PEB0122T ( = NCIMB 14961T = DSM 29779T).

Messiah

r1955, Boston.

Carbon sequestration and nutrient reserves under differen t land use systems

This study evaluated the contribution of agroforestry (AFS) and traditional systems to carbon sequestration and nutrient reserves in plants, litter and soil. The study was carried out in the semiarid region of Brazil in a long-term experiment on an experimental farm of the goat and sheep section of the Brazilian Agricultural Research Corporation (Embrapa). Two agroforestry systems were investigated: agrosilvopastoral (ASP) and forest-pasture areas (SP) as well as traditional agriculture management (TM), two areas left fallow after TM (six fallow years - F6 and nine fallow years - F9) and one area of preserved Caatinga vegetation (CAT). Soil, litter and plants were sampled from all areas and the contents of C, N, P, K, Ca and Mg per compartment determined. The AFS (ASP and SP) had higher nutrient stocks than the traditional and intermediate stocks compared to the preserved Caatinga. In the ASP, a relevant part of the nutrients extracted by crops is returned to the system by constant inputs of litter, weeding of herbaceous vegetation and cutting of the legume crops. After fallow periods of six and nine years, carbon and nutrient stocks in the compartments soil, litter and herbaceous plants were similar to those of the preserved Caatinga (CAT), but still lower than under natural conditions in the woody vegetation.

Tekijännimet Mikael Agricolan teosten kielessä. Henkilötarkoitteisten johdosten merkitykset, funktiot ja rakenteet

My research deals with agent nouns in the language of the works of Mikael Agricola (ca. 1510–1557). The main tasks addressed in my thesis have been to describe individual agent noun types, to provide a comprehensive picture of the category of agent nouns and to clarify the relations between different types of agent nouns. My research material consists of all the agent nouns referring to persons in the language of Agricola’s works, together with their context. The language studied is for the most part translated language. Agent nouns play an important role both in the vocabulary of natural language and in broader sentence structures, since in a text it is constantly necessary to refer to actors re-ferring to persons in the text. As a concept and a phenomenon, the agent noun is widely known in languages. It is a word formed with a certain derivational affixes, which typical-ly refers to a person. In my research the agent noun category includes both deverbal and denominal derivatives referring to persons, e.g. kirjoittaa > kirjoittaja (to write > writer), asua > asuva (to inhabit > inhabitant), imeä > imeväinen (to suck > suckling), juopua > juopunut (to drink > drunkard), pelätä > pelkuri (to fear > one who fears ‘a coward’), apu > apulainen (help/to help > helper); lammas > lampuri (sheep > shepherd). Besides original Finnish expressions, agent noun derivatives taken as such from foreign languages form a word group of central importance for the research (e.g. nikkari, porvari, ryöväri, based on the German/Swedish for carpenter, burgher, robber). Especially important for the formation of agent nouns in Finnish are the models offered by foreign languages. The starting point for my work is predominantly semantic, as both the criteria for collecting the material and the categorisation underlying the analysis of the material are based on semantic criteria. When examining derivatives, aspects relating to structure are also inevitably of central importance, as form and meaning are closely associated with each other in this type of vocabulary. The alliance of structure and meaning can be described in an illustrative manner with the help of structural schemata. The examination of agent nouns comprises on the one hand analysis of syntactic elements and on the other, study of cultural words in their most typical form. The latter aspect offers a research object in which language and the extralinguistic world, referents, their designations and cultural-historical reality are in concrete terms one and the same. Thus both the agent noun types that follow the word formation principles of the Finn-ish language and those of foreign origin borrowed as a whole into Finnish illustrate very well how an expression of a certain origin and formed according to a certain structural model is inseparably bound up with the background of its referent and in general with semantic factors. This becomes evident both on the level of the connection between cer-tain linguistic features and text genre and in relation to cultural words referring to per-sons. For example, the model for the designations of God based on agent nouns goes back thousands of years and is still closely linked in 16th century literature with certain text genres. This brings out the link between the linguistic feature and the genre in a very con-crete manner. A good example of the connection between language and the extralinguistic world is provided by the cultural vocabulary referring to persons. Originally Finnish agent noun derivatives are associated with an agrarian society, while the vocabulary relat-ing to mediaeval urbanisation, the Hansa trade and specialisation by trade or profession is borrowed and originates in its entirety from vocabulary that was originally German.

Anthrax in cattle in southern Brazil: 1978-2006

Ten outbreaks of anthrax occurred in cattle from 1978 to 2006 in southern Brazil, in 5 municipalities on the border with Uruguay, a country where the disease is frequent. The 10 outbreaks represented 0.2% of all bovine specimens received during the period by the Regional Diagnostic Laboratory of the Federal University of Pelotas, causing 267 deaths in a risk population of 6,605 head. The disease affected young and adult cattle mainly during summer. Only one farmer reported that sheep and horses were also affected. Clinically the peracute form was more frequent, but in some outbreaks the acute form with a clinical manifestation period of 6-48 hours was also observed. The source of infection was not established; but the reduced rainfall, associated with low, flat, flooded lands used for agriculture followed by animal grazing after harvest was probably related to the disease occurrence. Annual vaccination is an efficient way to prevent the disease.

Acute and chronic nervous signs in cattle associated with Phalaris angusta poisoning in Argentina

Phalaris angusta is a South American natural grass that produces poisoning in sheep and cattle in Argentina and Brazil. Phalaris spp. can produce unrelated forms of poisoning in ruminants, acute and chronic syndromes. The objective of this paper was to describe an outbreak of acute and chronic Phalaris nervous syndrome in 53 of 980 fattening steers and heifers in a farm of Buenos Aires province. On September of 2006 the animals developed nervous signs and died after 3-5 days. The herd was removed to a phalarisfree pasture. Three months later (on December) 15 new clinical cases developed in the herd. Necropsy performed in one affected calf showed neither grossly nor microscopic changes. Microscopically, there were no major alterations in tissues. Nervous signs had been described in some field cases where neither pigment deposition nor axonal degeneration could be detected. Clinical findings displayed by affected cattle after consumption of Phalaris angusta pastures resemble those observed by other authors in Phalaris staggers. This is the first report in Argentina where both syndromes were seen in the same herd.

Development of IgY antibodies in chickens and IgG in rabbits immunized against proteins of Pythium insidiosum isolated from horses in the state of Rio de Janeiro

Pythiosis is caused by Pythium insidiosum and the occurrence of disease in horses was described in the North and Northwest State of Rio de Janeiro, Brazil. The disease was described in cattle, sheep, humans, and horses in different states and regions across the country. This paper describes the development of IgY and IgG polyclonal antibodies, in chicken and rabbits, respectively against proteins extracted from kunkers and hyphae of P. insidiosum from affected horses. The proteins were recognized by chicken, rabbit and horse antibodies by immunodiffusion and Western blot against majority bands of 27 and 43 KDa, and titrated by ELISA. The antibodies IgY developed by the first time against Brazilian strains of P. insidiosum may represent a valuable tool in the detection of antigens of the pathogen and contribute to further studies aimed at immunotherapy and knowledge about this disease in endemic areas in Rio de Janeiro and in Brazil.

Brachiaria spp. poisoning of ruminants in Brazil

Brachiaria species are the most important grasses for cattle production in Brazil. However, a limiting factor for the use of Brachiaria spp. is their toxicity. Most outbreaks of hepatogenous photosensitization are caused by B. decumbens; however B. brizantha, B. humidicola and B. ruziziensis can also cause poisoning. The poisoning affects cattle, sheep, goats and buffalo. Sheep are more susceptible than other animal species and the young are more susceptible than adults. There are differences in susceptibility among animals of the same species and it has been suggested that this resistance is genetic. Also has been suggested that buffalo and probably some sheep are resilient, i.e. when poisoned these animals have histologic lesions and high GGT serum concentrations, but do not show clinical signs. In general, saponin concentrations are higher in growing plants, but outbreaks occur all over the year, probably due to unexplained rise in saponin concentration in the plant. A clinical syndrome of progressive weight loss and death, without photosensitization, has been reported in cattle poisoned by B. decumbens. Main preventive measures are based on the selection of resistant or resilient animals and on the development of Brachiaria species or varieties with low saponin concentration.

Effects of zearalenone in prepubertal gilts

Prepubertal gilts were fed with a diet containing zearalenone (ZEA) in a concentration of 0.75 mg/kg for 21 days. The effects of this mycotoxin on morphologic aspects of the reproductive tract as well as on complete blood count (CBC), serum biochemistry analysis (SBA) and humoral immune response against sheep red blood cells (SRBC) were evaluated. There was a significant increase (P<0.05) on the reproductive tract weight, vulvar area, height of the epithelial cells of endometrial glands and uterine mucosa. These results showed the ability of this nonsteroidal mycotoxin in mimicking actions of 17β estradiol at the concentration of 0.75mg/kg. No changes in weight gain, CBC, SBA parameters and humoral response against SRBC were observed.