Viability of sporulated oocysts of Neospora caninum after exposure to different physical and chemical treatments

The aim of the present study was to evaluate the viability of Neospora caninum sporulated oocysts after various chemical and physical treatments. Bioassays in gerbils and molecular techniques (PCR-RFLP) were used for identification of the oocysts shed by experimentally infected dogs. Sporulated oocysts were purified and divided into 11 treatment groups as follows: absolute ethanol for 1 hr; 20 C for 6 hr; 4 C for 6 hr; 60 C for 1 min; 100 C for 1 min; 10% formaldehyde for 1 hr; 10% ammonia for 1 hr; 2% iodine for 1 hr; 10% sodium hypochlorite for 1 hr; 70% ethanol for 1 hr; and one group was left untreated and kept as a positive control. All chemical treatments were performed at room temperature (37 C). A total of 33 gerbils, or 3 gerbils per treatment, were used for bioassays. After treatment, the oocysts were divided into aliquots of 1,000 oocysts and orally administered to gerbils. After 63 days, the gerbils were anesthetized and killed with 0.2 ml of T61; blood and tissue samples were collected for serological (IFAT and western blotting), molecular (real-time PCR), histopathology, and immunohistochemical tests. Treatments were considered effective only if all 5 detection techniques tested negative. High temperatures at 100 C for 1 min and 10% sodium hypochlorite for 1 hr were the only treatments that met this condition, effectively inactivating all oocysts.     

Diverse and atypical genotypes identified in Toxoplasma gondii from dogs in São Paulo, Brazil

The prevalence of Toxoplasma gondii in 118 unwanted dogs from S?o Paulo City, S?o Paulo State, Brazil, was determined. Antibodies to T. gondii were assayed by the modified agglutination test and found in 42 (35.8%) dogs, with titers of 1:20 in 10, 1:40 in 6, 1:80 in 5, 1:160 in 5, 1:320 in 6, 1:640 in 7, and 1:1,280 or higher in 3. Hearts and brains of 36 seropositive dogs were bioassayed in mice, or cats, or both. Tissues from 20 seropositive dogs were fed to 20 T. gondii-free cats. Feces of cats were examined for oocysts. Toxoplasma gondii was isolated from 15 dogs by a bioassay in mice, from the brain alone of 1, from the heart alone of 4, and from both brains and hearts of 10. All infected mice from 5 of 15 isolates died of toxoplasmosis during primary infection. Four additional isolates were obtained by bioassay in cats. Genotyping of these 19 T. gondii isolates using polymorphisms at 10 nuclear markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and a new SAG2 (an apicoplast marker Apico) revealed 12 genotypes. One isolate had Type III alleles at all 11 loci, and the remaining 18 isolates contained a combination of different alleles and were divided into 11 genotypes. The absence of Type II in Brazil was confirmed. The result supports previous findings that T. gondii population genetics is highly diverse in Brazil.     

Genome-wide association and biological pathway analysis for milk-fat composition in Danish Holstein and Danish Jersey cattle

Background

The milk fat profile of the Danish Holstein (DH) and Danish Jersey (DJ) show clear differences. Identification of the genomic regions, genes and biological pathways underlying the milk fat biosynthesis will improve the understanding of the biology underlying bovine milk fat production and may provide new possibilities to change the milk fat composition by selective breeding. In this study a genome wide association scan (GWAS) in the DH and DJ was performed for a detailed milk fatty acid (FA) profile using the HD bovine SNP array and subsequently a biological pathway analysis based on the SNP data was performed.

Results

The GWAS identified in total 1,233 SNPs (FDR < 0.10) spread over 18 chromosomes for nine different FA traits for the DH breed and 1,122 SNPs (FDR < 0.10) spread over 26 chromosomes for 13 different FA traits were detected for the DJ breed. Of these significant SNPs, 108 SNP markers were significant in both DH and DJ (C14-index, BTA26; C16, BTA14; fat percentage (FP), BTA14). This was supported by an enrichment test. The QTL on BTA14 and BTA26 represented the known candidate genes DGAT and SCD. In addition we suggest ACSS3 to be a good candidate gene for the QTL on BTA5 for C10:0 and C15:0. In addition, genetic correlations between the FA traits within breed showed large similarity across breeds. Furthermore, the biological pathway analysis revealed that fat digestion and absorption (KEGG04975) plays a role for the traits FP, C14:1, C16 index and C16:1.

Conclusion

There was a clear similarity between the underlying genetics of FA in the milk between DH and DJ. This was supported by the fact that there was substantial overlap between SNPs for FP, C14 index, C14:1, C16 index and C16:1. In addition genetic correlations between FA showed a similar pattern across DH and DJ. Furthermore the biological pathway analysis suggested that fat digestion and absorption KEGG04975 is important for the traits FP, C14:1, C16 index and C16:1.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-1112) contains supplementary material, which is available to authorized users.     

Efficiency of population structures for mapping of Mendelian and imprinted quantitative trait loci in outbred pigs using variance component methods

In a simulation study different designs for a pure line pig population were compared for efficiency of mapping QTL using the variance component method. Phenotypes affected by a Mendelian QTL, a paternally expressed QTL, a maternally expressed QTL or by a QTL without an effect were simulated. In all alternative designs 960 progeny were phenotyped. Given the limited number of animals there is an optimum between the number of families and the family size. Estimation of Mendelian and parentally expressed QTL is more efficient in a design with large family sizes. Too small a number of sires should be avoided to minimize chances of sires to be non-segregating. When a large number of families is used, the number of haplotypes increases which reduces the accuracy of estimating the QTL effect and thereby reduces the power to show a significant QTL and to correctly position the QTL. Dense maps allow for smaller family size due to exploitation of LD-information. Given the different possible modes of inheritance of the QTL using 8 to16 boars, two litters per dam was optimal with respect to determining significance and correct location of the QTL for a data set consisting of 960 progeny. The variance component method combining linkage disequilibrium and linkage analysis seems to be an appropriate choice to analyze data sets which vary in marker density and which contain complex family structures.     

Plant use in a city in Northern Italy during the late Mediaeval and Renaissance periods: results of the archaeobotanical investigation of “The Mirror Pit” (14th–15th century a.d.) in Ferrara

Results are presented from archaeobotanical analyses carried out in ‘The Mirror Pit’, a rectangular pit, situated in the city of Ferrara, in Emilia Romagna—Northern Italy. The study is part of wider research focused on the Mediaeval period of this city. The pit belongs to a building which was in use from the second half of 14th to the end of 15th century a.d. The pit itself was filled with domestic rubbish during a few years in the middle of the 15th century. The layers were preserved by waterlogging and are therefore extraordinarily rich in organic remains and artifacts, among which was the beautiful spherical mirror which gave its name to the pit. Approximately 256,000 well-preserved seeds and fruits belonging to 98 species or carpological types were identified in 12 l of sediment from the different layers. Ficus carica, Brassica rapa subsp. rapa/subsp. sylvestris and Vitis vinifera subsp. vinifera prevailed. Many cultivated plants and weeds were present, together with a smaller number of wild plants of wet environments and woodlands. Altogether they provided much new information on the diet (cultivated and wild fruits, cereals and pulses, vegetables, spices and medicinal/fibre/oil plants) of the middle to upper class inhabitants of a city in Northern Italy during the Renaissance period. The records also yielded evidence of domestic activities such as winemaking, oil making, mustard making and other kitchen practices.     

Synthesis and reactivity of diphosphino Pt(II) silanolate complexes as molecular models of the interaction of platinum particles with silica

A series of di- and monosubstituted cis-platinum(II) silanolate complexes, Pt(OSiR₃)₂(dppe) (R=Et, 1; R=Me, 2) and Pt(OSiR₃)Cl(dppe) (R=Et, 3; R=ⁱPr, 4) where dppe is 1,2-bis(diphenylphosphino)ethane, have been isolated and characterised spectroscopically. Complex 1 does not react with CO and H₂ under anhydrous conditions, but the complexes Pt{C(O)OCH₃)}₂(dppe) (6) and Pt(CO₃)(dppe) (7) have been isolated bubbling CO in methanol and CO₂ in moist benzene solutions of 1, respectively. The behaviour of 1 towards water or methanol is discussed on the basis of ¹H, and ³¹P{¹H} NMR spectroscopic data. The new complex Pt{S₂C(OSiEt₃)₂}(dppe) (8) has been isolated by reaction of 1 with CS₂ in benzene solution. This reactivity would suggest a high sensitivity towards water, but not towards H₂ or CO, of the bonding of slightly oxidised platinum particles with silanol groups of silica surface.     

The long history of Cannabis and its cultivation by the Romans in central Italy, shown by pollen records from Lago Albano and Lago di Nemi

The cores from the Albano and Nemi lakes, near Rome, were studied within the European Union funded PALICLAS project and provided high resolution records of the Late-glacial and Holocene. Pollen evidence of increasing human influence on vegetation was recorded in the Holocene parts of both diagrams, and the Cannabis (hemp) curve was one of the major signs. In this paper we present unambiguous pollen evidence from the Cannabaceae records for the cultivation of hemp in central Italy by the Romans. The oldest records of Cannabis and Humulus (hop) date from to the Late-glacial. Hop pollen values rise during the mid Holocene, while hemp pollen becomes more abundant from ca. 3000 cal B.P. onwards. The highest earliest hemp peak (21%) is dated to the 1st century A.D. This ‘Cannabis phase’, with the abrupt rise of hemp pollen soon after the rise of cultivated trees (Castanea, Juglans and Olea) is associated with the increase in cereals and ruderal plants. This unambiguous proof of cultivation by Romans around 2000 B.P. occurs as well as a long lasting pre-Roman presence of hemp in the area, which is natural and possibly also anthropogenic. Subsequent clear episodes of cultivation in the medieval period were found. Received February 4, 2002 / Accepted September 13, 2002 Correspondence to: Anna Maria Mercuri, e-mail: mercuri.annamaria@unimo.it     

拉萨高原居民肺容量的测定

对年龄、身高和体重相同的拉萨男性世居藏族39人和男性移居汉族43人的肺容量进行了测定。结果显示:藏族组的肺活量(VC)、肺总容量、胸围均大于汉族组,残气量有大于汉族组之趋势(P=0.06)。胸围的大小与VC呈正相关。5岁前和18岁后移居高原者之肺容量无差别。结果提示,拉萨世居藏族具有较大的肺容量,这对提高肺弥散功能和维持运动时的血氧饱和度有重要意义。     

A change in twist of actin provides the force for the extension of the acrosomal process in limulus sperm: the false-discharge reaction

One of the most spectacular motions is the generation of the acrosomal process in the limulus sperm. On contact with the egg, the sperm generates a 60-mum-long process that literally drills its way through the jelly surrounding the egg. This irresversible reaction takes only a few seconds. We suggested earlier that this motion is driven by a change in twist of the actin filaments comprising the acrosomal process. In this paper we analyze the so-called false discharge, a reversible reaction, in which the acrosomal filament bundle extends laterally from the base of the sperm and not anteriorly from the apex. Unlike the true discharge, which is straight, the false discharge is helical. Before extension, the filament bundle is coiled about the base of the sperm. In the coil, the bundle is not smoothly bent but consists of arms (straight segments) and elbows (corners) so that the coil looks like a 14-sided polygon. The extension of the false discharge works as follows: starting at the base of the bundle, the filaments change their twist which concomitantly changes the orientations of the elbows relative to each other; that is, in the coil, the elbows all like in a common plane, but after the change in twist, the plane of each elbow is rotated to be perpendicular to that of its neighbors. This change transforms the bundle from a compact coil into an extended left- handed helix. Because the basal end of the bundle is unconstrained, the extension is lateral. The true discharge works the same way but starts at the apical end of the bundle. The apical end, however, is constrained by its passage through the nuclear canal, which directs the extention anteriorly. Unlike the false discharge, during the true discharge the elbows are melted out, making the reaction irreversible. This study shows that rapid movement can be regenerated by actin without myosin and gives us insight into the molecular mechanism.     

Does menarche mark a period of elevated resting metabolic rate?

Resting metabolic rate (RMR) and body composition were measured in 44 initially nonoverweight girls at three time points relative to menarche: premenarche (Tanner stage 1 or 2), menarche (+/-6 mo), and 4 yr after menarche. Mean absolute RMR was 1,167, 1,418, and 1,347 kcal/day, respectively. Absolute RMR was statistically significantly higher at menarche than at 4 yr after menarche despite statistically significantly less fat-free mass (FFM) and fat mass (FM), suggesting an elevation in RMR around the time of menarche. The pattern of change in RMR, adjusted for FFM, log transformed FM, age, race, parental overweight, and two interactions (visit by parental overweight, parental overweight by FFM), was also considered. Adjusted RMR did not differ statistically between the visits for girls with two normal-weight parents. For girls with at least one overweight parent, adjusted RMR was statistically significantly lower 4 yr after menarche than at premenarche or menarche. Thus parental overweight may influence changes that occur in RMR during adolescence in girls.