Klebsiella to Salmonella gene transfer within rumen protozoa: implications for antibiotic resistance and rumen defaunation

The rumen has long been thought to be a site of gene transfer for microorganisms. Rumen protozoa (RPz) are active predators of bacteria that can harbor antibiotic resistance genes. In this study, RPz were assessed as sites of gene transfer between two bacterial species, Salmonella and Klebsiella. One Klebsiella isolate carried a plasmid bearing bla(CMY-2), encoding an extended-spectrum beta-lactamase conferring ceftriaxone resistance, while the Salmonella was susceptible to ceftriaxone yet capable of thriving within protozoa. In vitro studies revealed that ceftriaxone-resistant Salmonella could be isolated following co-incubation of Salmonella and Klebsiella with RPz obtained from adult cattle and goats. Ceftriaxone-resistant Salmonella were not recovered in the presence of an inhibitor of protozoa engulfment or when a protozoa-sensitive Salmonella was part of the co-incubation. This transfer event was additionally observed in vitro for protozoa-independent stressors although at a significantly lower frequency. The gene transfer event was related to bacterial conjugation since a conjugation inhibitor, nalidixic acid, perturbed the phenomenon. Ceftriaxone-resistant Salmonella were recovered from calves, sheep, and goats co-challenged with ceftriaxone-resistant Klebsiella and ceftriaxone-sensitive Salmonella. However, the transfer event was not observed in calves and sheep that were defaunated prior to the co-challenge. Moreover, Salmonella transconjugants were isolated from separate bovine in vivo studies involving a Klebsiella donor carrying a plasmid conferring colicin activity while no such transconjugants were obtained from defaunated calves. These results provide an important basis for evaluating and preventing the spread of antibiotic resistance and other selective advantages for pathogens present in ruminants.     

Clovis age Western Stemmed projectile points and human coprolites at the Paisley Caves

The Paisley Caves in Oregon record the oldest directly dated human remains (DNA) in the Western Hemisphere. More than 100 high-precision radiocarbon dates show that deposits containing artifacts and coprolites ranging in age from 12,450 to 2295 (14)C years ago are well stratified. Western Stemmed projectile points were recovered in deposits dated to 11,070 to 11,340 (14)C years ago, a time contemporaneous with or preceding the Clovis technology. There is no evidence of diagnostic Clovis technology at the site. These two distinct technologies were parallel developments, not the product of a unilinear technological evolution. "Blind testing" analysis of coprolites by an independent laboratory confirms the presence of human DNA in specimens of pre-Clovis age. The colonization of the Americas involved multiple technologically divergent, and possibly genetically divergent, founding groups.     

Visual Colour Shades in Pelts from Brown Mink (Mustela vison) Explained Colourimetrically

Microspectrophotometric methods were used to provide objective correlates to the visually judged colour shade (CS) of the underfur in scanbrown mink pelts. The study included 21 scanbrown mink pelts (winter coat), representing a larger group of 87 mink pelts coming from a feeding trial with experimental and control animals. The pelts were visually graded from 1 (blue-greyish) to 5 (reddish), primarily in respect of the CS (in auction classification=clarity of colour) of the underfur seen on the edge of the pelt. Prepared samples of underfur fibres showed in visual inspection a gliding change of colour from paler to darker in a proximal-distal direction and were consequently examined at three levels above the skin surface. The means of the dominant wavelengths (hues) were 589.40, 588.00 and 586.70 nm, respectively, indicating that the variation between levels was small. Investigation of the small pelt material concerned showed that CS was significantly correlated with the measured lightness (L*) and with the yellow chromaticity coordinate (b*) of the underfur samples. In this investigation it was also shown that the underfur fibres with a visually blue-greyish colour shade were relatively dark and less yellow compared with underfur fibres with a reddish colour shade. However, data and the model estimating the colour shade demonstrated that, for the material concerned, visually very reddish underfur colour shades were not found in the lightest coloured pelts.     

Low compliance with recommendations on folic acid use in relation to pregnancy: is there a need for fortification?

OBJECTIVE: As a means to prevent neural tube defects (NTDs), women planning pregnancy in Denmark are recommended to take a dietary supplement of 400 microg folic acid daily during the periconceptional period. We examined compliance with this recommendation in a national survey. DESIGN: Cohort study on pregnant women in Denmark. SETTING: The Danish National Birth Cohort (DNBC). SUBJECTS: From November 2000 to February 2002, 22,000 pregnant women were recruited for DNBC. Use of dietary supplements was recorded at enrollment. Compliance with the recommendation was related to an information campaign that took place during the second half of 2001, and to lifestyle factors provided in a telephone interview by the end of the first trimester of pregnancy. RESULTS: An increase was seen in the proportion of women complying with the recommendation in the study period and this coincided with the information campaign events. However, even at the end of the period, only 22.3% of the women who had planned their pregnancy fully complied with the recommendation. No increase at all was seen in periconceptional folic acid use among women with unplanned pregnancies. Young age, low education and smoking were identified as factors that determined non-compliance. CONCLUSIONS: Alternative and more effective strategies are needed if the Danish population is to benefit fully from the knowledge that folic acid prevents NTDs. Future strategies should not only target vulnerable groups, such as the less educated and the young, but also women who get pregnant without planning this. The only possible way to reach the last group may be through fortification of foods with folic acid.     

西藏绒山羊羊绒细度候选基因筛选及其chi-miR-105a靶基因的验证

为研究西藏绒山羊羊绒细度,以及了解相关候选基因对绒山羊产绒性能的影响,本研究对筛选chi-miR-105a靶基因进行验证,以西藏绒山羊为研究对象,联合RNA测序数据和蛋白质组学数据,筛选与羊绒细度相关的关键候选基因,并通过实时荧光定量和双荧光素酶报告基因试验对候选基因进行调控作用验证。结果表明:1)通过转录组和蛋白组数据整理得到384个DE mRNAs,12个DE miRNAs和29个DEPs。通过多组学联合分析发现CXCL10、SRC、CXCL9、FOS、ETV6、GMPS和PIP5K1B基因与羊毛细度相关。2)通过DEG-DE miRNA互作网络图发现chi-miR-105a与靶基因ETV6和PIP5K1B均在网络当中出现。因此对该调控因子进行验证和分析,根据靶基因表达水平试验发现转染chi-miR-105a mimics后引起毛乳头细胞中ETV6基因的mRNA表达显著降低(P<0.001),而PIP5K1B基因的表达量均无显著变化。3)通过双荧光素酶报告基因试验表明,过表达chi-miR-105a后,ETV6酶活性显著降低,即chi-miR-105a可与ETV6的3′UTR区结合。综上,通过多组学联合分析筛选出与羊毛细度相关基因CXCL10、SRC、CXCL9、FOS、ETV6、GMPS和PIP5K1B。通过双荧光素酶报告基因进行chi-miR-105a与其预测靶基因ETV6和PIP5K1B的靶标验证,确定chi-miR-105a是ETV6潜在的调控因子,本研究为加快优质绒山羊新品种(系)的培养提供理论依据。     

A spatial-temporal model of cell activation

A spatial-temporal model of calcium messenger function is proposed to account for sustained cellular responses to sustained stimuli, as well as for the persistent enhancement of cell responsiveness after removal of a stimulus, that is, cellular memory. According to this model, spatial separation of calcium function contributes to temporal separation of distinct phases of the cellular response. At different cellular sites, within successive temporal domains, the calcium messenger is generated by different mechanisms and has distinct molecular targets. In particular, prolonged cell activation is brought about by the interaction of calcium with another spatially confined messenger, diacylglycerol, to cause the association of protein kinase C with the plasma membrane. Activity of the membrane-associated protein kinase C is controlled by the rate of calcium cycling across the plasma membrane. In some instances, a single stimulus induces both protein kinase C activation and calcium cycling and thus causes prolonged activation; but in others, a close temporal association of distinct stimuli brings about cell activation via interaction of these intracellular messengers. Persistent enhancement of cell responsiveness after removal of stimuli is suggested to be due to the continued association, or anchoring, of protein kinase C to the membrane.     

Aerial Transmission of Thielaviopsis basicola, a Pathogen of Corn-Salad, by Adult Shore Flies

ABSTRACT Chlamydospores of Thielaviopsis basicola were consistently observed in frass excreted by adults and larvae of shore flies that were collected in the immediate vicinity of naturally infected corn-salad plants obtained from a commercial greenhouse production facility. Approximately 95% of the adult flies and 85% of the larvae were internally infested with the pathogen. Pathogen-free adult shore flies were subsequently shown to acquire the pathogen by ingestion after feeding on naturally infected plants. Viable propagules of the pathogen were excreted by these internally infested adults and were capable of transmitting the pathogen to healthy seedlings, which subsequently became infected.     

Investigations on the Influence of Helminth Parasites on Vaccination of Chickens against Newcastle Disease Virus under Village Conditions

Prevalence studies have shown that almost 100% of free-range chickens are infected with a wide range of parasites. The infections are mostly subclinical in nature, resulting in production losses and occasionally mortality. Newcastle disease (ND), on the other hand, results in high mortality rates during epidemics. ND is a limiting factor for increasing poultry production in many tropical countries, where frequent reports indicate vaccination failures. The aim of our study was to investigate the influence of helminths on the antibody response after vaccination against Newcastle disease of free-range chickens naturally infected with parasites. Sixty chickens were divided into six groups, of which three were vaccinated against ND with a live De Soto vaccine, while the other three remained non-vaccinated. One group within the vaccinated groups and the one within the non-vaccinated group was kept naturally infected with helminth parasites, while the other two groups in each set were dewormed with fenbendazole and niclosamide, and one of each of these groups was subsequently infected with Ascaridia galli. After vaccination, all the groups were followed for 5 weeks and their antibody titres were determined weekly using a HI test. All the birds were finally challenged 4 weeks after vaccination with a virulent velogenic ND virus obtained from a field outbreak. All the vaccinated chickens seroconverted and had high antibody levels after 3 weeks, but these dropped to low levels at 4 weeks after vaccination. After challenge, the antibody titres rose in the dewormed groups but not in the parasite-infected groups. After 5 weeks, all the parasite-infected animals had significantly lower antibody titres than the dewormed animals. All the vaccinated chickens survived the challenge infection, emphasizing the importance of the cellular immune response. Further studies are needed to examine the effects of the parasitic infection on protection against ND over a longer period.     

Cytotoxic constituents of Alectra and Striga species

Decimation of cereal growth and yields by hemiparasitic Striga species cannot be accounted for entirely by the removal of host‐plant resources. The production of toxic compounds by the parasite has been suggested. An investigation of three species of the economically important Striga and the related Alectra vogelii has now resulted in the isolation of eight iridoid glucosides (mussaenosidic acid, mussaenoside, gardoside methyl ester, bartsioside, isoaucubin, melittoside, aucubin and eurostoside), two caffeoyl phenylethyl glycosides (calceolarioside  A and verbascoside) as well as shikimic acid and trigonelline, all identified by NMR spectroscopy. The iridoids are potent cytotoxins and probably represent an anti‐herbivore defence system common to Scrophulariaceae (sensu lato). This has the potential to explain differences in tolerance apparent for contrasting host taxa and cultivars. The nature of the iridoids present also provides additional validation of the recent transferral of parasitic Scrophulariaceae (s.l.) to Orobanchaceae.     

Survival of sea‐water‐adapted trout,Salmo trutta L. ranched in a Danish fjord

The effect of seawater adaptation on the survival of coastally released post‐smolt trout, Salmo trutta L., was investigated by release: (1) directly (with no adaptation); (2) after retention in net pens in the sea for 29–131 days (delayed release); (3) after feeding with a high‐salt diet (12–13.5% NaCl) for 4 weeks; and (4) after a combination of (2) and (3). In total, 17 640 trout (age = 1+, 1.5 and 2+ years; mean fork lengths = 18.2–25.6 cm) were released in 14 batches in the summer or autumn months of 1986–1989. All fish were of domesticated origin and Carlin tagged. Survival and instantaneous mortality rates (total and fishing mortality) were estimated from reported recaptures. Mortality rates were estimated for: (1) the post‐smolt period; (2) the period until the legal size of capture (40 cm) was attained; and (3) for larger sea‐trout. Release with a delay of 4 weeks gave an increased survival rate. A longer adaptation period did not increase survival. On average, survival was increased by 36%. Survival was not increased by high‐salt diets. Until attainment of the legal size for capture, survival was 9.6% higher on average, with extremes as low as 1.7% and as high as 38% in individual batches.