Academic literature on the topic 'Non-ruminant research'

The fate of transgenic DNA (tDNA) and protein from feed derived from Genetically Modified organisms (GMOs) in animals has been a major issue since their commercialization in 1996. Several studies have investigated the risks of horizontal gene transfer (HGT) of tDNA and protein to bacteria or animal cells/tissues, but some of the reported data are controversial. Previous reports showed that tDNA fragments or proteins derived from GM plants could not be detected in tissues, fluids, or edible products from livestock. Other researchers have shown that there is a possibility of small fragments entering animal tissues, fluids and organs. This motivated us to update our knowledge about these concerns. Therefore, this review aimed to evaluate the probable transfer and accumulation of tDNA/proteins from transgenic feeds in animal samples (ruminant and non-ruminant) by evaluating the available experimental studies published scientifically. This study found that the tDNA/protein is not completely degraded during feed processing and digestion in Gastro-Intestinal Tract (GIT). In large ruminants (cattle), tDNA fragments/proteins were detected in GIT digesta, rumen fluid, and faeces. In small ruminants (goats), traces of tDNA/proteins were detected in GIT digesta, blood, milk, liver, kidney, heart and muscle. In pigs, they were detected in blood, spleen, liver, kidney, and GIT digesta. In poultry, traces were detected in blood, liver and GIT digesta but not in meat and eggs. Notwithstanding some studies that have shown transfer of tDNA/protein fragments in animal samples, we cannot rely on these few studies to give general evidence for transfer into tissues/fluids and organs of farm animals. However, this study clearly shows that transfer is possible. Therefore, intensive and authentic research should be conducted on GM plants before they are approved for commercial use, investigating issues such as the fate of tDNA or proteins and the effects of feeding GM feed to livestock.

Improving plant cell wall (fiber) degradation remains one of the highest priority research goals for all ruminant enterprises dependent on forages, hay, silage, or other fibrous byproducts as energy sources, because it governs the provision of energy-yielding nutrients to the host animal. Although the predominant species of microbes responsible for ruminal fiber degradation are culturable, the enzymology and genetics underpinning the process are poorly defined. In that context, there were two broad objectives for this proposal. The first objective was to identify the key cellulosomal components in Ruminococcus albus and to characterize their structural features as well as regulation of their expression, in response to polysaccharides and (or) P AA/PPA. The second objective was to evaluate the similarities in the structure and architecture of cellulosomal components between R. albus and other ruminal and non-ruminal cellulolytic bacteria. The cooperation among the investigators resulted in the identification of two glycoside hydrolases rate-limiting to cellulose degradation by Ruminococcus albus (Cel48A and CeI9B) and our demonstration that these enzymes possess a novel modular architecture specific to this bacterium (Devillard et al. 2004). We have now shown that the novel X-domains in Cel48A and Cel9B represent a new type of carbohydrate binding module, and the enzymes are not part of a ceiluiosome-like complex (CBM37, Xu et al. 2004). Both Cel48A and Cel9B are conditionally expressed in response to P AA/PPA, explaining why cellulose degradation in this bacterium is affected by the availability of these compounds, but additional studies have shown for the first time that neither PAA nor PPA influence xylan degradation by R. albus (Reveneau et al. 2003). Additionally, the R. albus genome sequencing project, led by the PI. Morrison, has supported our identification of many dockerin containing proteins. However, the identification of gene(s) encoding a scaffoldin has been more elusive, and recombinant proteins encoding candidate cohesin modules are now being used in Israel to verify the existence of dockerin-cohesin interactions and cellulosome production by R. albus. The Israeli partners have also conducted virtually all of the studies specific to the second Objective of the proposal. Comparative blotting studies have been conducted using specific antibodies prepare against purified recombinant cohesins and X-domains, derived from cellulosomal scaffoldins of R. flavefaciens 17, a Clostridium thermocellum mutant-preabsorbed antibody preparation, or against CbpC (fimbrial protein) of R. albus 8. The data also suggest that additional cellulolytic bacteria including Fibrobacter succinogenes S85, F. intestinalis DR7 and Butyrivibrio fibrisolvens Dl may also employ cellulosomal modules similar to those of R. flavefaciens 17. Collectively, our work during the grant period has shown that R. albus and other ruminal bacteria employ several novel mechanisms for their adhesion to plant surfaces, and produce both cellulosomal and non-cellulosomal forms of glycoside hydrolases underpinning plant fiber degradation. These improvements in our mechanistic understanding of bacterial adhesion and enzyme regulation now offers the potential to: i) optimize ruminal and hindgut conditions by dietary additives to maximize fiber degradation (e.g. by the addition of select enzymes or PAA/PPA); ii) identify plant-borne influences on adhesion and fiber-degradation, which might be overcome (or improved) by conventional breeding or transgenic plant technologies and; iii) engineer or select microbes with improved adhesion capabilities, cellulosome assembly and fiber degradation. The potential benefits associated with this research proposal are likely to be realized in the medium term (5-10 years).

The key objectives of this BARD project were to: (1) study long-term effects of immunization of prepubertal ewes against recombinant ovine placental lactogen (roPL) on subsequent birth weights of their lambs and their milk production; (2) optimize the anti-roPL immunization protocol using adjuvant preparations acceptable to producers and regulatory agencies; and (3) determine the physiological mechanism(s) whereby immunization against oPL increases fetal growth and development and mammogenesis. These objectives were based on key findings from a previous BARD project that: (a) immunization of ewes against roPL increased lamb birth weight and ewe milk production during lactation; (b) roPL and recombinant ovine growth hormone (roGH) increased the proliferation and differentiated function of endometrial glands that, in turn, would enhance uterine secretions necessary for fetal and placental growth; and (c) exogenous roPL and roGH stimulated mammogenesis and milk production during lactation. The BARD projects address central problems in sheep production, including reproductive failure due to embryonic/fetal mortality, low birth weight of lambs especially in prolific breeds, and reduced milk yields which affect neonatal survival. The sheep placenta secretes both lactogenic (oPL) and somatogenic (oGH) hormones. The receptors for those hormones are present in the fetus and placenta as well as maternal uterus, and mammary gland. Our research has focused on determining the biological role of these placental hormones in development and differentiation of the uterus during gestation and the mammary gland during pregnancy and lactation. Studies conducted in the current BARD project indicated that the effects of anti-roPL immunization were variable in ewes and that commercially available and widely acceptable adjuvant preparations were not effective to produce high anti-roPL titers in pre-pubertal ewes. In the non-prolific Rambouillet ewe in Texas and in the Awassi and the Assaf in Israel, anti-roPL immunization increased lamb birth weight; however, the magnitude of this effect and the inherent variability precluded our ability to determine the physiological mechanism of how the immunization increases fetal growth. Collectively, our findings suggest that anti-roPL immunization is not currently feasible as an easy and efficacious tool for the producer to increase flock reproductive and production efficiency. The variability in response of individual ewes to anti-roPL immunization likely includes modifying the recombinant hormone and the type of adjuvant used for the immunization. In particular, the oPL may need to be modified to ensure maximum antigenicity in a broad range of breed types. Nonetheless, the investigators continue to collaborate on identifying fundamental mechanisms that can be improved by genetics or management to enhance the efficiency of uteroplacental function and, in turn, fetal growth and development. High prolificacy is a desirable trait in intensive sheep production systems. One of the main limitations of using prolific breeds of sheep is that increased litter size is associated with low birth weights and increased mortality of lambs. Further, low birth weight is associated with an increased propensity for adult diseases and decreased production efficiency. Indeed, our recent studies find that the birth weights of lambs born in large litters can be improved by both genetics and management. Future cooperative research will continue to focus on reproductive efficiency of sheep that have broader implications for improving production efficiency in all types of ruminant livestock.

This research project was performed in context of the apparent probiotic effect of selected lactic acid bacteria (LAB) silage inoculants on the performance of ruminants (improved feed intake, faster live-weight gain, higher milk yields and improved feed efficiency). The overall objective was to find out how LAB affect ruminant performance. The project included several “chapters” as follows: 1. The effect of LAB silage inoculants on the survival of detrimental bacteria in rumen fluid, in vitro study (Weinberg et al., The Volcani Center). An in vitro model was developed to study the interaction between selected LAB and an E. coli strain tagged with green fluorescence protein (GFP) in buffered RF. Results indicated that both LAB inoculants and E. coli survived in the RF for several days; both LAB inoculants and LAB-treated silages did not affect survival of E. coli in rumen fluid in vitro. The effect of feeding baled wheat silages treated with or without three selected LAB silage inoculants on the performance of high-lactating cows (Weinberg et al., The Volcani Center). Treatments included control (no additive), Lacobacillusbuchneri40788 (LB), Lactobacillus plantarumMTD1 40027 (LP) and Pediococcuspentosaceus30168 (PP), each applied at 10⁶ cfu/g FM. The silages were included in the TMR of 32 high milking Holstein cows in a controlled feeding experiment. All baled silages were of good quality. The LB silage had the numerically highest acetic acid and were the most stable upon aerobic exposure. The cows fed the LB silages had the highest daily milk yields, percent milk fat and protein. The microbiome of baled wheat silages and changes during ensiling of wheat and corn (Sela et al., The Volcani Center). Bacterial community of the baled silages was dominated mainly of two genera in total, dominated by Lactobacillus and Clostridium_sensu_stricto_12 with 300 other genera at very low abundance. Fungal community was composed mainly of two genera in total, dominated by Candida and Monascuswith 20 other genera at very low abundance. In addition, changes in the microbiome during ensiling of wheat and corn with and without addition of L. plantarumMTD1 was studied in mini-silos. Overall 236 bacterial genera were identified in the fresh corn but after 3 months Lactobacillus outnumbered all other species by acquiring 95% of relative abundance. The wheat silage samples are still under analysis. The effect of applying LAB inoculants at ensiling on survival of E. coli O157:H7 in alfalfa and corn silages(Adesogan et al., University of Florida). E. coli (10⁵ cfu/g) was applied to fresh alfalfa and corn at ensiling with or without L. plantarumor L. buchneri. The pathogen was added again after about 3 moths at the beginning of an aerobic exposure period. The inoculants resulted in faster decrease in pH as compared with the control (no additives) or E. coli alone and therefore, the pathogen was eliminated faster from these silages. After aerobic exposure the pathogen was not detected in the LAB treated silages, whereas it was still present in the E. coli alone samples. 5. The effect of feeding corn silage treated with or without L. buchnerion shedding of E. coli O157:H7 by dairy cows (Adesogan et al., UFL). BARD Report - Project 4704 Page 2 of 12 Five hundred cows from the dairy herd of the University of Florida were screened for E. coli shedding, out of which 14 low and 13 high shedders were selected. These cows were fed a total mixed ration (TMR) which was inoculated with E. coli O157:H7 for 21 days. The TMR included corn silage treated with or without L. buchneri. The inoculated silages were more stable upon aerobic exposure than the control silages; the silage inoculant had no significant effect on any milk or cow blood parameters. However, the silage inoculant tended to reduce shedding of E. coli regardless of high or low shedders (p = 0.06). 6. The effect of feeding baled wheat silages treated with or without three selected LAB silage inoculants on the rumen microbiome (Mizrahi et al., BGU). Rumen fluid was sampled throughout the feeding experiment in which inoculated wheat silages were included in the rations. Microbial DNA was subsequently purified from each sample and the 16S rRNA was sequenced, thus obtaining an overview of the microbiome and its dynamic changes for each experimental treatment. We observed an increase in OTU richness in the group which received the baled silage inoculated with Lactobacillus Plantarum(LP). In contrast the group fed Lactobacillus buchneri(LB) inoculated silage resulted in a significant decrease in richness. Lower OTU richness was recently associated in lactating cows with higher performance (Ben Shabatet al., 2016). No significant clustering could be observed between the different inoculation treatments and the control in non metric multi-dimentional scaling, suggesting that the effect of the treatments is not the result of an overall modulation of the microbiome composition but possibly the result of more discrete interactions. Significant phylum level changes in composition also indicates that no broad changes in taxa identity and composition occurred under any treatment A more discrete modulation could be observed in the fold change of several taxonomic groups (genus level analysis), unique to each treatment, before and after the treatment. Of particular interest is the LB treated group, in which several taxa significantly decreased in abundance. BARD Report - Project 4704 Page 3 of 12