Design of a specific colonic mucus marker using a human commensal bacterium cell surface domain

Imaging living cells and organs requires innovative, specific, efficient, and well tolerated fluorescent markers targeting cellular components. Such tools will allow proceeding to the dynamic analysis of cells and the adaptation of tissues to environmental cues. In this study, we have identified and synthesized a novel non-toxic fluorescent marker allowing a specific fluorescent staining of the human colonic mucus. Our strategy to identify a molecule able to specifically bind to the human colonic mucus was on the basis of the mucus adhesion properties of commensal bacteria. We identified and characterized the mucus-binding property of a 70-amino acid domain (MUB(70)) expressed on the surface of Lactobacillus strains. The chemical synthesis of MUB(70) was achieved using the human commensal bacterium Lactobacillus reuteri AF120104 protein as a template. The synthesized Cy5-conjugated MUB(70) marker specifically stained the colonic mucus on fixed human, rabbit, and guinea pig tissues. Interestingly, murine tissue was not stained, suggesting significant differences in the composition of the murine colonic mucus. In addition, this marker stained the mucus of living cultured human colonic cells (HT29-MTX) and human colonic tissue explants. Using a biotinylated derivative of MUB(70), we demonstrated that this peptide binds specifically to Muc2, the most abundant secreted mucin, through its glycosylated moieties. Hence, Cy5-MUB(70) is a novel and specific fluorescent marker for mammalian colonic mucus. It may be used for live imaging analysis but also, as demonstrated in this study, as a marker for the diagnosis and the prognosis of colonic mucinous carcinomas.     

The effects of feed restriction during organogenesis on embryo-fetal development in the rat

BACKGROUND: Given the role of nutrition and body weight gain in normal development, pharmaceuticals intended to reduce appetite and promote weight loss will generate safety data that may be challenging to interpret. To aid with this, the effects of feed restriction and subsequent body weight reductions on embryo-fetal development were investigated in the rat. METHODS: Groups of 20 timed pregnant female Sprague-Dawley rats were offered Certified Rodent Diet 5002 either ad libitum or in restricted amounts of 20, 15, 10, and 7.5 g/day from Gestation Day (GD) 6-17. Clinical signs, body weights, and food consumption were recorded. Cesarean sections were performed on GD 21 and fetuses were sexed, weighed, and examined for external, visceral, and skeletal development. RESULTS: Mean maternal body weights at the end of the feed restriction period, GD 18, were reduced 0.87 x, 0.80 x, 0.69 x, and 0.63 x control mean in the 20, 15, 10, and 7.5 g/day groups, respectively. Mean body weight gains for the restriction period inclusive, GD 6-18, were 0.49 x and 0.24 x control at 10 and 7.5 g/day, respectively, and a mean body weight loss occurred at 10 and 7.5 g/day (0.95 x and 0.85 x mean GD 6 body weight, respectively). Fetal body weights were reduced 0.95 x, 0.93 x, 0.90 x, and 0.76 x control at 20, 15, 10, and 7.5 g/day, respectively. This resulted in a reduction in gravid uterine weight at 10 and 7.5 g/day. There were no external, visceral, or skeletal malformations attributed to feed restriction. There was an increase in the skeletal variation of wavy ribs and a decrease in ossification at 7.5 g/day. CONCLUSIONS: These data demonstrate that feed restriction-induced reductions in maternal gestational body weight gain of approximately 50% compared to ab lib fed rats only caused a reduction in fetal body weight. Even up to a 15% maternal gestational body weight loss had no effect on embryo viability in rats, but retarded fetal growth significantly enough to induce minor changes in skeletal development. There were no external, visceral, or skeletal malformations associated with any of the levels of maternal body weight reduction or loss.     

Correlation between the immune response elicited in rabbits by env-recombinant avipox vaccines and the inhibition of HIV-1-specific functions

The fine immunoreactivity of the rabbit humoral response elicited by four env-recombinant avipoxviruses and their ability to stimulate a memory T-cell response and a protective immunity have been studied. The antibody specificity was compared with the serum neutralizing activity and virus-specific T-cell proliferative response. Resistance to challenge by cell-associated HIV-1 was monitored by PCR. Canarypox (CP) and fowlpox (FP) constructs, containing the complete env gene (IS(+)) from the HIV-1(SF2) strain, induced a higher profile of epitope recognition than their counterparts expressing the env gene deleted of the putative immunosuppressive region (IS(-)). Serum neutralizing activity was in agreement with fusion inhibition and lymphoproliferative response in rabbits immunized with CPIS(+), and only partially with FPIS(+). Rabbits failed to be infected, but anti- p55 gag-specific antibodies could be demonstrated by Western blot. This study confirms the ability of these non-replicative live recombinant viruses to elicit a complete immune response, capable of inhibiting specific HIV-1 functions.     

Genetic markers on BTA14 predictive for residual feed intake in beef steers and their effects on carcass and meat quality traits

With the high cost of feed for animal production, genetic selection for animals that metabolize feed more efficiently could result in substantial cost savings for cattle producers. The purpose of this study was to identify DNA markers predictive for differences among cattle for traits associated with feed efficiency. Crossbred steers were fed a high‐corn diet for 140 days and average daily feed intake (ADFI), average daily gain (ADG), and residual feed intake (RFI) phenotypes were obtained. A region on chromosome 14 was previously associated with RFI in this population of animals. To develop markers with the highest utility for predicting an animal's genetic potential for RFI, we genotyped additional markers within this chromosomal region. These polymorphisms were genotyped on the same animals (n = 1066) and tested for association with ADFI, ADG and RFI. Six markers within this region were associated with RFI (P ≤ 0.05). After conservative correction for multiple testing, one marker at 25.09 Mb remained significant (P = 0.02) and is responsible for 3.6% of the RFI phenotypic variation in this population of animals. Several of these markers were also significant for ADG, although none were significant after correction. Marker alleles with positive effects on ADG corresponded to lower RFI, suggesting an effect increasing growth without increasing feed intake. All markers were also assessed for their effects on meat quality and carcass traits. All of the markers associated with RFI were associated with adjusted fat thickness (AFT, P ≤ 0.009) and three were also associated with hot carcass weight (HCW, P ≤ 0.003). Marker alleles associated with lower RFI were also associated with reduced AFT, and if they were associated for HCW, the effect was an increase in weight. These markers may be useful as prediction tools for animals that utilize feed more efficiently; however, validation with additional populations of cattle is required.     

Active polyphenolic compounds,nutrient contents and antioxidant capacity of extruded fish feed containing purple coneflower (Echinacea purpurea (L.) Moench.)

The growth of fish is directly dependent on feed composition and quality. Medicinal plants can be added to fish feed as adjuvant therapy for the prevention of fish diseases. The purple coneflower (Echinacea purpurea (L.) Moench.) has been reported to have multiple biological effects, including immunomodulatory and antioxidant activity. The most active compounds of E. purpurea are polyphenols - caffeic acid derivatives: caftaric acid, chlorogenic acid, cynarin, echinacoside and cichoric acid.Due to a relatively limited number of studies on the use of the purple coneflower as a nutritional supplement for fish feeding, extruded fish feed with addition of Echinacea roots was produced. In the feed total phenolic content, selected polyphenol contents, the energetic value, nutrient contents and antioxidant capacity were examined.The results indicate that fish feed with addition of the Echinacea has a great potential to be a good source of natural radical scavengers, for example polyphenols, and nutritive ingredients. Antioxidant properties of feed were well correlated with the coneflower content. The study findings confirmed that high-temperature extrusion-cooking process does not deactivate phenolic antioxidant compounds, which are present both in the Echinacea roots and in the final product. Fish feed with addition of E. purpurea can be used as a nutritional supplement in the prevention of fish diseases caused by oxidative stress.     

DNase I and Proteinase K eliminate DNA from injured or dead bacteria but not from living bacteria in microbial reference systems and natural drinking water biofilms for subsequent molecular biology analyses

Molecular techniques, such as polymerase chain reaction (PCR) and quantitative PCR (qPCR), are very sensitive, but may detect total DNA present in a sample, including extracellular DNA (eDNA) and DNA coming from live and dead cells. DNase I is an endonuclease that non-specifically cleaves single- and double-stranded DNA. This enzyme was tested in this study to analyze its capacity of digesting DNA coming from dead cells with damaged cell membranes, leaving DNA from living cells with intact cell membranes available for DNA-based methods. For this purpose, an optimized DNase I/Proteinase K (DNase/PK) protocol was developed. Intact Staphylococcus aureus cells, heat-killed Pseudomonas aeruginosa cells, free genomic DNA of Salmonella enterica, and a mixture of these targets were treated according to the developed DNase/PK protocol. In parallel, these samples were treated with propidium monoazide (PMA) as an already described assay for live-dead discrimination. Quantitative PCR and PCR-DGGE of the eubacterial 16S rDNA fragment were used to test the ability of the DNase/PK and PMA treatments to distinguish DNA coming from cells with intact cell membranes in the presence of DNA from dead cells and free genomic DNA. The methods were applied to three months old autochthonous drinking water biofilms from a pilot facility built at a German waterworks. Shifts in the DNA patterns observed after DGGE analysis demonstrated the applicability of DNase/PK as well as of the PMA treatment for natural biofilm investigation. However, the DNase/PK treatment demonstrated some practical advantages in comparison with the PMA treatment for live/dead discrimination of bacterial targets in drinking water systems.     

Bioavailability of two organic forms of zinc in comparison to zinc sulphate for weaning pigs fed a diet composed mainly of wheat,barley and soybean meal

This study was performed to compare the bioavailability of two organic zinc compounds, a zinc glycinate complex and a zinc amino acid chelate with that of zinc sulphate in growing pigs fed a basal diet composed mainly of wheat, barley and soybean meal. The experiment included 96 pigs with an average body weight of 8 kg, allotted to ten groups of nine to ten pigs each. The first group received the basal diet, containing 42 mg of native zinc per kg, without zinc supplementation over a period of five weeks. The other nine groups received the basal diet supplemented with 15, 30 or 50 mg of zinc/kg as zinc sulphate, zinc glycinate or the zinc amino acid chelate. Pigs fed the unsupplemented diet had a lower growth performance (body weight gain, feed conversion ratio) than the other nine groups. Supplementation of 15 mg zinc/kg diet (irrespective of zinc form) was sufficient to yield optimum growth performance. Plasma zinc concentration and activity of alkaline phosphatase were rising with increasing zinc supplementation levels up toa maximum reached at a supplementary level of 30 or 50 mg/kg diet for activity of alkaline phosphatase and plasma zinc concentration, respectively. The response of those parameters to zinc supplementation did, however, not differ between thethree zinc compounds considered. The apparent digestibility of zinc from the diet was also not different for the three zinc compounds. In conclusion, these findings show that the bioavailability of the two organic zinc compounds did not differ from that of zinc sulphate in growing pigs fed a diet with wheat, barley and soybean meal as major components.     

Design a system for measuring individual cow feed intake in commercial dairies

Monitoring individual cow feed intake is necessary for calculating the cow individual feed efficiency. The cost and maintenance time necessary for research systems make them impractical for most of the commercial producers. We developed a measurement system with producer convenience and low investment as key design criteria. The goal of this study was to design the system and validate its ability to rank cows by their feed conversion efficiency in commercial farms. The new system consisted of three principal parts: (a) a hanging weighing system, (b) a visual cow identification system and (c) an automatic cleaning system. The weighing system consisted of hanging a single load cell to provide feed mass measurements. The image-based cow identification system (replacing Radio-Frequency Identification) entailed cameras installed above the feeding area and an image processing algorithm that recognized cows by their collar numbers. The new system worked within normal farm routines: the feed supplying truck distributed the animal feed, and a tractor cleaned feed residual. To validate the accuracy and convenience of the system and to rank the cows by their efficiency, an experiment with six scales and 12 cows was conducted in a research barn, succeeded by eight-scale system in a commercial farm with 16 cows. The feed intake of each cow participating in the experiments was monitored for one month. The validation experiment showed that the system had the following specification: scales were accurate within 120 g; the visual cow identification rate was greater than 96%; feeding duration was accurate to 52 s; and routine farm practices (feed distribution, pushing, and residual removal) continued as usual. The cost for a feeding station (utilized consequently for a number of cows) was about 1 500 USD. An example of application of the system to rank cows by their efficiency under commercial conditions was shown. The system can potentially be used for ranking cows by their efficiency in commercial facilities.     

蛹虫草饲料添加剂的研究现状及展望

蛹虫草饲料添加剂包括蛹虫草子实体、蛹虫草培养残基、蛹虫草及其培养残基提取物、蛹虫草菌固液发酵产物、微生物发酵蛹虫草残基等产品.蛹虫草饲料添加剂含有粗蛋白、粗脂肪、氨基酸等营养成分,以及虫草素、腺苷、多糖等活性成分,在畜禽、反刍动物、水产品等动物养殖中的应用均获得较好的效果.对蛹虫草子实体、蛹虫草培养残基、蛹虫草及其培养...